Kyushu University Academic Staff Educational and Research Activities Database
List of Papers
YUTAKA BANNO Last modified date:2020.06.18

Professor / Genetic resources / Department of Bioscience and Biotechnology / Faculty of Agriculture

1. Eiichi Nagashima, Masao Nakagaki, Masaaki Sato, Hidetada Wakabayashi, Yutaka Banno, Zhonghuai Xiang, Testosteron-like immunoreactivities in the haemolymph of the silkworm, Bombyx mori, Journal of Insect Biotechnology and Sericology, 10.11416/kontyushigen1930.52.243, 52, 3, 243-244, 1983.01.
2. Yutaka Banno, Yutaka Kawaguchi, Meng Kui Xu, Hiroshi Doira, Inheritance of female specific protein in larval hemolymph of Bombyx mori, Journal of Insect Biotechnology and Sericology, 10.11416/kontyushigen1930.53.549, 53, 6, 549-550, 1984.
3. Hiroshi Doira, Hajime Kihara, Yutaka Banno, Genetical studies on the non-molting dwarf” mutation in Bombyx mori, Journal of Insect Biotechnology and Sericology, 10.11416/kontyushigen1930.53.427, 53, 5, 427-431, 1984.02, The recessive lethal mutation “non-molting dwarf” (symbol, nm-d) was found in F2progenies of the cross between a female sparsely mottled with fine translucent dots and a normal male. Linkage test showed that the nm-d gene was linked to Dominant chocolate (I-a) which had been mapped at position 5.9 on the ninth linkage group. Precise localization of nm-d was performed by a three-point experiment of the type I I-a/nm-d sib-mating. The locus of I (Yellow inhibitor) gene on the genetic map corresponds to position 0.0. The recombination value between I and nm-d was calculated to be 0.13%, nm-d and I-a 6.60%, I and I-a 6.74%, respectively, through the observation of 10,895 individuals. Hence the arrangement of these three gene loci on the chromosome is in the order of I/nm-d-I-a, and nm-d gene lies at position 0.1 on the ninth linkage group..
4. Yutaka Banno, Yutaka Kawaguchi, Hiroshi Doira, Sexual dimorphism and developmental changes in hemolymph amylase of Bombyx mori, Journal of Insect Biotechnology and Sericology, 10.11416/kontyushigen1930.53.335, 53, 4, 335-340, 1984.02, The amylases in hemolymph of Bombyx mori were assayed by Somogyi-Nelson's reduction method and by starch-iodine reaction after polyacrylamide gel electrophoresis. Total enzyme activities were plotted against stages of development and metamorphosis from the third larval instar to emergence of moth. The maximum amylase activity was shewn at the end of the fifth larval instar, and sexual dimorphism in the activity was defined. When activity was expressed in maltose units/ml hemolymph, females had a greater activity than males in the later half of the fifth instar to the middle stage of pupal life throughout. Different electrophoretic banding patterns for amylases were produced as a function of development and sexuality. Hemolymph amylases were separated into eleven active bands, only the band c was detected in common throughout the developmental stages tested. Band 1 was shown to appear at the middle stage of the fifth instar and disappeared in pupal hemolymph. Bands S-l to S-4 were detected in female larvae of the fifth instar, but not in pupal hemolymph nor in male ones. Band SV was identified characteristically of female pupae. Electrophoretic mobilities of these female-specific amylases coincided with those of female-specific hemclymph proteins thus far recognized by dye-staining..
5. Yutaka Banno, Yutaka Kawaguchi, Hiroshi Doira, Genetical studies on the N-methyl-N-nitrosourea induced non-molting b“ mutation in Bombyx mori”, Journal of Insect Biotechnology and Sericology, 10.11416/kontyushigen1930.54.227, 54, 3, 227-231, 1985, Newly laid silkworm eggs of a inbred normal strain were immersed in 0.5mM N-methyl-N-nitrosourea solution for 60 minutes. The eggs were at an age of 90±15 minutes when the treatment was started. In the M3 progenies after the treatment, a considerable number of dwarf larva was found on the second day of the first larval instar. All dwarf larvae continued to eat mulberry leaves slightly increasing body-size for about ten days, and then died without entering into molt. Duration of the first larval instar is 3–4 days for normal development of this insect. Genetic analysis revealed that the dwarf and non-molting lethal character was controlled by an induced recessive mutation. The mutant was named “non-molting b” with gene symbol nm-b. Linkage experiment showed that nm-b was linked with plain (p) and Yellow-blood (Y) which had been mapped at 0.0 and 25.6 position, respectively, on ganetic map of the second linkage group. Precise localization of nm-b gene was made by a three-point experiment involving pM, Y and nm-b. The recombination value between pMand Y was calculated to be 25.68%, nm-b and Y 0.55%, and pMand Y 26.23%, Hence the arrangement of these three gene loci on the chromosome is in. the order of p-nm-b-Y. Taking correction factor into account, the locus of of nm-b is determined as 25.1 position on the second linkage group..
6. Yutaka Kawaguchi, Hiroshi Doira, Yutaka Banno, Hiroshi Fujii, Biological effects of N-methyl-N-nitrosourea as revealed by soaking of newly laid eggs of Bombyx mori, Journal of Insect Biotechnology and Sericology, 10.11416/kontyushigen1930.54.213, 54, 3, 213-221, 1985.01, Egg shell of Bombyx mori is porous and having many spiracles on the surface, chemical compounds of low molecular weight can pass through the shell and penetrate into egg-plasm. Present experiment concerns with MNU, a known mutagenic and carcinogenic agent that react directly with DNA and protein in living organisms. Eggs of an inbred normal strain of Bombyx were immersed in aqueous solution of MNU for 15 minutes at the age of 0.5 hr to 24 hr. Eggs at 0.5 hr age undergo meiotic division and syngamy of egg and sperm-nuclei takes place around 2 hr after oviposition, followed by synchronous cleavage mitosis with cycle 60 minutes. Gastrulation takes place at egg age 24 hr. Effect on fertilization rate was negligible after MNU-treatment at 0.5 hr age. The effectiveness of the treatment was determined by killing of embryos. Hatchability was extremely low when treated at early stages, from meiosis to second cleavage division, then increased rapidly and attained normal survival level at 8. 5 hr and thereafter treatment with MNU. Formation of cellular membrane surrounding cleavage nucleus migrated to periplasm was observable in 9. 5 hr egg. MNU-treatment also caused various anomalies in hatched larvae including malformations and monstrosities. Frequency of induced larval anomalies changing with the progress of developmental stages at the treatment was in parallel with that of killing effect. Soaking of egg in MNU solution effectively induced mutation, we discovered several mutants in descending progenies of the treatment..
7. Yutaka Kawaguchi, Yutaka Banno, Toru Shimada, Masahiko Kobayashi, Linkage studies on the Storage protein-2 gene (Pst) in Bombyx mori, Journal of Insect Biotechnology and Sericology, 10.11416/kontyushigen1930.55.243, 55, 3, 243-245, 1986.01, Codominant manifestation of the gene controlling electrophoretic mobility of storage protein-2 in larval haemolymph of Borribyx mori was reconfirmed, and the gene was named “Storage protein-2” with symbol Pst. Precise localization of Pst was made by a three-point experiment involving lem and Ze. The gene lem have been mapped at 0.0 position and Ze at 20.8, respectivelly, on the third linkage group. The recombination value between lem and Pst was 15.25%, Pst-Ze 3.71% and lem-Ze 18.64%. Taking correction factor into account, the locus of Pst was determined to be 16.7 to the right of lem and 4.1 to the left of Ze. Hence the Pst gene is localized at 16.7 on genetic map of the third linkage group..
8. Mitsuiku Nakayama, Hiroshi Doira, Yutaka Banno, Revision of the genetic map of the linkage group, Journal of Insect Biotechnology and Sericology, 10.11416/kontyushigen1930.57.53, 57, 1, 53-56, 1988, It was revealed that the mutant Nd-t, which had been mapped at 0.0 position on the twenty-third linkage group, was not a gene mutation but a chromosomal aberration of the type T (23; 25) Nd. A series of crossing over experiments was carried out to revise the genetic map of the twenty-third linkage group. Heterozygous F1 males of the type BphB tub sp/Bphc ++ were backcrossed to BphB tub sp females. The recombination value between Bph and tub obtained from the segregation in male larvae was 6.88%, whereas the value between tub and sp was calculated to be 15.98% based on the observation of female moths. Taking previously determined order of gene arrangement on the chromosome into consideration, the map units of the genes on the twenty-third linkage group were revised as follows..
9. Yutaka Banno, Hiromu Akai, Ultrastructural changes of corpus allatum in the imaginal stages in the silkworm, Bombyx mori, Journal of Insect Biotechnology and Sericology, 10.11416/kontyushigen1930.57.431, 57, 5, 431-437, 1988, Ultrastractural changes of the corpus allata during the imaginal stages were observed light and electron microscopically. Several characteristic changes of the corpus allata were found with the progress of time after the emergence. However, the ultrastructural differences of corpus allata between male and female moths were not markedly detected. The results obtained were as follows: 1) From just after the emergence, a large number of small vacuole are detected in the cytoplasm. After the emergence these vacuoles gradually increased in both number and size, occupying a great part of their cytoplasm. 2) The neurosecretory axons containing electron dense glanules are observed in both conective tissue envelope and cytoplasmic area. These granules increased from the 24th to 72nd hrs after the emergence, while the vacuoles increased in these axons. 3) Large numbers of mitochondria distribute in the cytoplasm, and their criste are clearly detected just after the emergence. These structures are however, disappeared gradually. 4) The nucleus contains many chromatin masses and nucleoli without any sign of synthetic activity. 5) The lysosome increases in the cytoplasm, and the basemembrane disorganized. These results indicate that the corpus allatum is clearly in the process of the histolysis during the imaginal stage..
10. Yutaka Kawaguchi, Yutaka Banno, Hiroshi Doira, Hiroshi Fujii, Manifestation of Characteristics in the Giant Egg “Mutant of Bombyx mori (Lepidoptera
Bombycidae). 2. Artificial Induction of Large Eggs by Application of Large Amounts of 20-hydroxyecdysone.”, Japanese Journal of Applied Entomology and Zoology, 10.1303/jjaez.33.63, 33, 2, 63-68, 1989.01, Injection of large amounts of 20-hydroxyecdysone into normal female pupae of Bombyx mori altered egg production. Large eggs were produced in addition to normal eggs. 1) These eggs were as large and heavy as Ge mutant eggs. 2) Yolk protein contents in these large eggs had almost the same values as those of Ge eggs. However, there is no qualitative difference in the protein patterns of 20-hydroxyecdysone induced large eggs and normal eggs. 3) Large eggs were induced by injection of 20-hydroxyecdysone at doses of more than 50 μg. 4) Induction of large eggs was achieved at the expense of the total number of eggs, similarly to the case of Ge mutants. 5) Distribution of the enlarged eggs in a ovariole was not at random, but clustered in definite regions..
11. Yutaka Banno, Yutaka Kawaguchi, Hiroshi Doira, Shinji Tochihara, Hemolymph proteins during larval development of bombyx mori
Proteins of young larvae, Journal of Insect Biotechnology and Sericology, 10.11416/kontyushigen1930.62.187, 62, 3, 187-194, 1993, Hemolymph proteins were analyzed quantitatively and qualitatively throughout the larval development of Bombyx mori. The hemolymph protein concentration decreased by the 4th larval instar, with cyclic increases until molting and decreases after ecdysis. The protein concentration increased rapidly after the 4th molt and reached a maximum value at the end of the 5th instar, with female larvae showing higher concentration than the male ones. This sexual dimorphism was obtained only after day 2 of the 5th instar. The electrophoretic analysis of hemolymph proteins revealed the existence of two groups of major proteins from hatching to the 3rd larval instar. Their content decreased gradually in the 4th instar and they disappeared in the 5th instar. The two groups should be referred to as “Proteins of young larvae -A and -B (PYL-A and PYL-B)”. PYL-A and PYL-B consisted of four and three proteins, respectively. In the late 5th instar, hemolymph proteins consisted of 3 major proteins (SP-2, 30kDa proteins and FL-1 with the latter being minor in the male) and many minor ones. These results clearly indicated that characteristics of the hemolymph at the young larval stage (1st instar to 3rd instar) were very different from those of the 5th instar in terms of the major protein and total protein concentrations..
12. Yutaka Kawaguchi, Yutaka Banno, Katsumi Koga, Hiroshi Doira, Hiroshi Fujii, Characteristic profiles of haemolymph proteins during larval development of molting mutants of the silkworm, Bombyx mori, Comparative Biochemistry and Physiology - B Biochemistry and Molecular Biology, 10.1016/0305-0491(93)90242-W, 105, 2, 361-367, 1993.01, 1. 1. Electrophoresis of the haemolymph from developing larvae of Bombyx mori revealed that the banding patterns of proteins could be grouped into three characteristic types. 2. 2. The young larval type exhibited at the first to the third instars (which are the phase of nutritional growth). 3. 3. The late larval type specific to the fifth instar (which is the phase of reproductive growth). 4. 4. The intermediary type shown at the fourth instar (which is the stage of switchover between the above two phases). 5. 5. The tetramolter mutant M3 lacked the intermediary type (i.e. the feature of the fourth instar) during its development, whereas the different tetramolter mutant rt had a mixture of the late and the intermediate types at its fourth (last) instar. 6. 6. On the other hand, the mutant M5, which is a pentamolter, repeated the intermediary type at its fourth and fifth (penultimate) instars..
13. Kazuhiro Fujikawa, Yutaka Kawaguchi, Yutaka Banno, Hiroshi Doira, Katsumi Koga, Inheritance of a new mutant, “shirotae-ran” in Bombyx mori, Journal of Insect Biotechnology and Sericology, 10.11416/kontyushigen1930.62.88, 62, 1, 88-90, 1993.01.
14. Yutaka Kawaguchi, Yutaka Banno, Katsumi Koga, Hiroshi Doira, Hiroshi Fujii, Polygonal patterns on eggshells of giant egg mutant and large eggs induced by 20-hydroxyecdysone in Bombyx mori, Journal of Insect Physiology, 10.1016/0022-1910(93)90032-M, 39, 5, 437-443, 1993.01, The polygonal network patterns, i.e. the secretory surface imprints produced by the follicular epithelial cells, on the lateral side region of eggshell in Bombyx mori were observed by scanning electron microscopy and differential interference contrast microscopy in order to characterize the giant egg (Ge) mutant and large eggs induced by the injection of 20-hydroxyecdysone into pupae. The area of each polygon was smaller in Ge but larger in the induced large eggs than in the normal eggs. The polygon number per unit area was much higher in Ge than in the normal eggs, but the induced large eggs exhibited the same number as the normal eggs. These results indicate that Ge is a mutant in which the number of the follicular epithelial cells is increased, whereas injected 20-hydroxyecdysone promoted hypertrophy of the follicle cells during choriogenesis..
15. Banno Y., Kawaguchi Y. and Doira H., Cytogenetical analysis of chromosomal aberration due to translocation between the 23rd and 25th linkage groups in the silkworm, Bombyx mori., Hereditas, 10.1111/j.1601-5223.1993.00259.x, 118, 3, 259-263, 118(3),259-263, 1993.01.
16. Zhe Qi Meng, Yutaka Banno, Katsumi Koga, Yutaka Kawaguchi, Yu Yi Lu, Effects of NaF on the calcium ion concentration in isolated midgut cells of Bombyx mori, Journal of Insect Biotechnology and Sericology, 10.11416/kontyushigen1930.63.310, 63, 4, 310-314, 1994, The free Ca2+ concentration in midgut cells isolated from fifth instar larvae of Bombyx mori was found to be 592 nM as measured by addition of fura-2/AM to the culture medium. When the cultured cells were loaded with NaF during the measurement, the Ca2+ concentration decreased to half the original value in 90 sec. This effect of NaF was completely blocked when verapamil, a competitor for the membrane transport system, was added, indicating that free Ca2+ was transported out of the cells after the addition of NaF. This finding may be of importance from the viewpoint of the protection of sericulture from polluted air containing fluoride compounds..
17. Yutaka Kawaguchi, Yutaka Banno, Katsumi Koga, Hiroshi Doira, Hiroshi Fujii, Haemolymph protein profiles in allatectomy-induced trimolters resemble those of a recessive trimolting mutant of the silkworm, Bombyx mori, Comparative Biochemistry and Physiology - B Biochemistry and Molecular Biology, 10.1016/0305-0491(94)90188-0, 107, 4, 579-584, 1994.01, Two types of artificial trimolters named A and B were induced by excising the corpora allata from the normal tetramolter of Bombyx mori at the periods before and after, respectively, the third molt. By using polyacrylamide gel electrophoresis, these trimolters were analysed for the banding patterns of haemolymph proteins at their final (fourth) instar. A feature resembling that of the last (fifth) instar of the normal tetramolter, i.e. with an increasing difference in content of the female-specific protein (FL), was found to occur. However, this was preceded by a feature of the penultimate instar of the normal tetramolter, i.e. with a low content of haemolymph proteins without sexual differences. The trimolter-B exhibited less distinct differences in FL content than the trimolter-A. These patterns of haemolymph proteins in the allatectomy-induced trimolters were similar to those of the recessive trimolter mutant rt..
18. Yutaka Kawaguchi, Yutaka Banno, Katsumi Koga, Hiroshi Doira, Hiroshi Fujii, Induction of larger eggs by 20-hydroxyecdysone in giant egg mutant phenotypes in the silkworm, Bombyx mori, Journal of Insect Biotechnology and Sericology, 10.11416/kontyushigen1930.63.443, 63, 6, 443-448, 1994.01, The influence of exogenous 20-hydroxyecdysone on the giant egg (Ge) mutant of Bombyx mori was investigated. The Ge female pupae injected with the hormone produced larger and heavier eggs than the control Ge pupae injected with water alone, and these characters of the eggs depended upon the dose of the hormone. The area of each polygon, which is observed on the eggshell surface of the enlarged Ge egg, was broader than that of polygon on the control Ge egg, while the total polygon number per whole lateral region remained unchanged. These results indicate that the administration of 20-hydroxyecdysone promoted hypertrophy, without increase in number, of the follicular epithelial cells which undergo choriogenesis. The Ge eggs thus modified would be the largest of the hitherto reported eggs of B. mori..
19. Banno Y., Noda K., Kawaguchi Y., Koga K. and Doira H., Linkage studies on a hemolymph protein named "Protein of young larval-A4 (PYL-4)" of the silkworm, Bombyx mori., J. Seric. Sci. Jpn., 63(4), 299-302, 1994.01.
20. Yutaka Kawaguchi, Kazutaka Kawakami, Hiroshi Doira, Yutaka Banno, Katsumi Koga, Genetic analysis of the soft and elongated trunk mutant in Bombyx mori, Journal of Insect Biotechnology and Sericology, 10.11416/kontyushigen1930.64.31, 64, 1, 31-34, 1995, The dominant mutant “Soft and elongated trunk” (symbol Set) was found as an MNU-induced artificial gene mutation, which has been inherited in a sex-linked manner. It is characterized by the elongated 6th-7th body segments, making the body shape abnormally stretched. Hemizygous female individuals with respect to this gene are completely lethal at the embryonic stage. The Set gene was localized at the 35.5 position of the first linkage group on the basis of a three-point experiment involving os and sch as marker genes..
21. Kazuhiro Fujikawa, Yutaka Kawaguchi, Yutaka Banno, Katsumi Koga, Hiroshi Doira, Yolk of a "scanty vitellin" mutant, vit, of Bombyx mori is lacking in vitellin and 30 kDa proteins, Comparative Biochemistry and Physiology -- Part A: Physiology, 10.1016/0300-9629(95)02031-4, 112, 3-4, 585-589, 1995.01, A novel mutant named "scanty vitellin" (vit) of Bombyx mori, induced by N-methyl-N-nitrosourea, was analyzed for yolk proteins by polyacrylamide gel electrophoresis in the presence of sodium dodecyl sulfate. The ovary of vit/vit pupae was lacking in vitellin and the 30 kDa proteins that are yolk constituents synthesized outside the ovary. However, the vit/vit ovary had a normal amount of the egg-specific protein, a yolk component synthesized by the ovarian follicular epithelial cells. On the other hand, vitellogenin and the 30 kDa proteins, which are the precursors of yolk proteins, were abundant in the haemolymph of vit/vit female pupae. These results indicate that the mutant gene interferes with the uptake of the extraovarian yolk precursors into the oocyte..
22. Banno Y., Kawaguchi Y., Koga K. and Doira H., Genetical studies on the "non-molting k" mutant in the silkworm, Bombyx mori., 64(3), 219-223, 1995.01.
23. Banno Y., Kawaguchi Y., Koga K. and Doira H., Postreductional meiosis revealed in males of the mutant with chromosomal aberration "T (23;25)Nd" of the silkworm, Bombyx mori., J. Seric. Sci. Jpn., 64(5), 410-414, 1995.01.
24. Yutaka Kawaguchi, Yutaka Banno, Katsumi Koga, Hiroshi Doira, Changes of yolk and haemolymph proteins during ovarian development in the miniature egg mutant, emi, of Bombyx mori, Journal of Insect Biotechnology and Sericology, 10.11416/kontyushigen1930.65.13, 65, 1, 13-20, 1996, The changes in the amount of yolk and haemolymph proteins were investigated during ovarian development in a mutant which produces small sized eggs and was named the miniature egg (emi). The ovary of this mutant possessed not only vitellin (H and L) and the 30 kDa proteins synthesized in extra-ovarian tissues but also the egg specific protein (ESP) synthesized in the follicular epithelial cells of the ovary. However, the contents of these proteins in the mutant ovary were considerably lower than in the normal ovary. On the other hand, large amounts of vitellogenin and the 30 kDa proteins were found to remain in the haemolymph of emi females at the late pupal and the adult stages. The emi gene is likely to affect neither incorporation of vitellogenin and the 30 kDa proteins into the ovary nor synthesis of ESP in the ovary. There is the possibility that the gene is expressed in the process of determination of egg size..
25. Yutaka Kawaguchi, Keiko Sumida, Yutaka Banno, Katsumi Koga, Hiroshi Doira, Features and growth of the stemmata in the varnished eye mutant of Bombyx mori, Journal of Insect Biotechnology and Sericology, 10.11416/kontyushigen1930.65.441, 65, 6, 441-446, 1996, The varnished eye mutant (ve) of the silkworm, Bombyx mori exhibited undeveloped ommatidia in the adult compound eyes. However, the morphology and growth rate of the stemmata, a larval optic organ located on both sides of the head capsule in 6 pairs, were completely normal in developing ve larvae. Also the sectional structures of the stemmata observed under a light microscope did not show any abnormality in the mutant. These results suggest that the formation and growth of larval stemmata are not affected by the ve mutant gene..
26. Kazuhiro Fujikawa, Yutaka Kawaguchi, Yutaka Banno, Katsumi Koga, Hiroshi Doira, Morphological Analysis of Eggshell in the Shirotae-ran Mutant, vit, Bombyx mori, Journal of the Faculty of Agriculture, Kyushu University, 41, 1-2, 75-81, 1996, The morphology of the eggshell (chorion) of the shirotae-ran mutant, vit (wherein the yolky substances are poor), of Bombyx mori was observed by scanning electron microscopy and differential interference contrast microscopy. The egg size was slightly smaller in the mutant than in the normal. The surface structures of the micropyle in the anterior pole region and the basic patterns of the network structures (polygons) imprinted on the lateral flat surfaces as well as the mode of distribution of the small knobs in the posterior, ventral and dorsal regions exhibited no peculiarity in the mutant. The polygons were significantly smaller in size in the mutant than in the normal. However, the number of polygons per whole lateral side did not differ between the mutant and the normal. The cross section of the mutant chorion was as usual in terms of layer structure and thickness. These findings imply that the choriogenesis occurs ordinarily in the vit mutant..
27. Sakaida K., Banno Y., Nakamura T., Kawaguchi Y., Doira H.,and Koga K., Aberrant meiotic behavior of chromosomes due to reciprocal translocation in the EDs mutant of the silkworm, Bombyx mori., Hereditas, 10.1111/j.1601-5223.1996.00025.x, 125, 1, 25-29, 125 (1), 25-29, 1996.01.
28. Song F., Banno Y., Kawaguchi Y., Koga K. and Xiang Z., Dimorphism in chromosome number of Samia cynthia walkeri in China., Int. J. Wild Silkworm & Silk, 2, 11-13, 1996.01.
29. Banno Y., Kawaguchi Y., Koga K. and Doira H., Chromosomal features at different meiotic stages during spermatogenesis in Bombyx mori., Sericologia, 36(1), 51-57, 1996.01.
30. Yutaka Kawaguchi, Yutaka Banno, Katsumi Koga, Takeshi Kawarabata, Hiroshi Doira, Comparsion of Chorion Structure of Eggs from Bombyx mori, Bombyx mandarina (Lepidoptera
Bombycidaee) and Their First Filial Generation, Applied Entomology and Zoology, 31, 3, 408-415, 1996.08.
31. Hidenao Yamada, Yutaka Banno, Katsumi Koga, Hiroshi Doira, Genetic analysis of the “translucent-15” mutant in Bombyx mori, Journal of Insect Biotechnology and Sericology, 10.11416/kontyushigen1930.66.113, 66, 2, 113-115, 1997, a recessive mutation was found spontaneously in the f11 strain maintained in Kyushu University. It had characteristics of moderately translucent or oily larval skin. Linkage analysis showed that this gene was linked to the bl gene. The novel mutant gene was named translucent-15 (symbol oft). It was localized at position 42.7 centimorgans on the 15th linkage group on the basis of a three-point experiment involving bl and Se as markers..
32. Ko Zo Tsuchida, Yutaka Banno, Kazuo Hashid, Methods for fluorescence in situ hybridization using oocyte and spermatocyte chromosomes of Bombyx mori, Journal of Insect Biotechnology and Sericology, 10.11416/kontyushigen1930.66.233, 66, 4, 233-241, 1997.
33. Yutaka Banno, Kosaku Sakaida, Takashi Nakamura, Yutaka Kawaguchi, Katsumi Koga, Hiroshi Doira, Kozo Tsuchida, Reassessment of mapping of the E homeotic gene complex of Bombyx mori by linkage analysis and in situ hybridization with an Antennapedia clone as a probe, Journal of Insect Biotechnology and Sericology, 10.11416/kontyushigen1930.66.151, 66, 3, 151-155, 1997.01, The gene for a recessive mutant named “soft and limp” (gene symbol sol) was shown to be linked with the E homeotic pseudoalleles previously mapped to the distal end of the 6th linkage group. A three-point test involving one of the E genes (Eca) and the F gene localized on the same linkage group indicated that the arrangement of these three gene loci is in the order of sol—Eca—F. Taking the recombination values and correction factors into account, the locus of the sol gene was determined to be -21.1 cM of the 6th linkage group. These results suggest that the 0.0 position of this linkage group should be the sol gene instead of the E loci. This inference was supported by the results of fluorescence in situ hybridization with an RNA probe derived from the B. mori Antennapedia gene closely linked with the E complex loci; the signal was detected at an internal region of a chromosome, not at its distal end..
34. BANNO Y., GOTO H., YASUOKA M., NAKAMURA T., KAWAGUCHI Y., KOGA K.and Doira H, Genetic studies on the sex linked non-molting mutaition, nm-s, of Bombyx mori. , J.Seric.Sci.Jnp., 66(5):351-355, 1997.01.
35. Yutaka Kawaguchi, Shunsuke Akagi, Yutaka Banno, Katsumi Koga, Eiichi Kuwano, Hiroshi Doira, Protein Profiles of Larval Haemolymph of Bombyx mori in Artificially Induced Trimolting larvae using a 1,5-disubstituted Imidazole, Journal of the Faculty of Agriculture, Kyushu University, 42, 1-2, 203-209, 1997.12, Trimolter larvae (undergoing three larval molts) of Bombyx mori were induced artificially by administrating 1-benzyl-5-[(E)-2,6)-dimethyl-1,5-heptadienyl]-imidazole (an insect growth regulator called KK-42) shortly after the third molt, and analyzed by polyacrylamide gel electrophoresis for the banding patterns of haemolymph proteins. The features were similar to those previously reported for allatectomy-induced trimolters, which mimicked the trait of the recessive trimolting mutant rt rather than that of the dominant trimolting mutant M3..
36. Y. Kawaguchi, Masataka Iwakuma, Yutaka Banno, K. Koga, Alteration of egg character in the wemi mutation of Bombyx mori by the application of 20-hydroxyecdysone, Journal of Insect Biotechnology and Sericology, 67, 1, 31-35, 1998, The influence of exogenous 20-hydroxyecdysone on the expression of the miniature egg (emi) mutation of Bombyx mori was investigated. Female pupae of emi injected with the hormone produced larger and heavier eggs than the inherent emi eggs. The enlarged emi eggs were approximately similar to the normal eggs, indicating that the administration of 20-hydroxyecdysone recovered the inferiority of the emi character..
37. Yutaka Kawaguchi, Katsumi Koga, Yutaka Banno, Hlroshi Doira, Inheritance and characteristics of a novel low-temperature sensitive gray-egg mutation in Bombux mori, Journal of Insect Biotechnology and Sericology, 10.11416/kontyushigen1930.67.265, 67, 4, 265-270, 1998, A recessive mutation affecting the eggshell character was found spontaneously in some strains stocked in Kyushu University. It had characteristics of the so-called “gray egg”, wherein the eggshell is opaque or white to various degrees. This trait was expressed only when female pupae were chilled at 8 to 10°C for 24hr (usually to suppress pupal-adult development). Control animals continuously kept at 25°:C did not produce such eggs. The ratio of cold-induced gray eggs to the total eggs was about 7% per batch. Linkage analysis using one the sensitive strains (cll) showed that the gene controlling this character was linked to the ok gene which belongs to the 5th linkage group. The novel mutant gene was named the “low-temperature sensitive gray-egg” with the gene symbol tsg..
38. Yutaka Kawaguchi, Takahiro Kusakabe, Katsumi Koga, Yutaka Banno, Effects of ooplasmic size on the larval development as observed in the small egg mutant emi of Bombyx mori, Journal of Insect Biotechnology and Sericology, 10.11416/kontyushigen1930.68.245, 68, 3, 245-250, 1999.
39. Takashi Nakamura, Yutaka Banno, Yutaka Kawaguchi, Katsumi Koga, Improvement of mating method for the wild silkmoth, Bombyx mandarina, Journal of Insect Biotechnology and Sericology, 10.11416/kontyushigen1930.68.165, 68, 2, 165-166, 1999.01.
40. Nakamura, T., Banno, Y., and Fujii, H., Genetics of the “wild silkworm translucent” mutant (ows), discovered in the progenies after cross between the domesticated silkworm, Bombyx mori and the
wild mulberry silkworm, Bombyxmandarina, Int. J. Wild silkmoth & silk, 6, 7-10, 1999.01.
41. NAKAMURA T., ,BANNO Y., NAKADA T., NHO S. K., XU M. K., UEDA K.,KAWARABATA, Geographic dimorphism of the wild silkworm,Bombyx mandarina, in the chromosome number and the occurrence of the retroposon-like insertion in the arylphorin gene, Genome, 10.1139/gen-42-6-1117, 42, 6, 1117-1120, 42(6),1117-1120, 1999.01.
42. Yonghuang Jiang, Yutaka Banno, Hiroshi Fujii, Genetic variant and inheritance of the 35K protease in digestive juice of silkworm, Bombyx mori, Journal of Insect Biotechnology and Sericology, 10.11416/kontyushigen1930.69.225, 69, 3, 225-227, 2000.01.
43. Yonghuang Jiang, Koji Shirai, Toshihisa Okido, Yutaka Banno, Hiroshi Fujii, Purification of 35K protease from the digestive juice of Bombyx mori, Journal of Sericultural Science of Japan, 10.11416/kontyushigen1930.69.47, 69, 1, 47-53, 2000.01, 35K protease was purified from the digestive juice of Bombyx mori by a series of chromatography using Butyl-Toyopearl, Sephadex G-50 and DEAE-Sephacel. The purified protease gave a single protein band with a molecular mass of 35,000 on SDS-PAGE and its pi was 9. 1. It had optimal activity at pH11 and in the temperature under 40°C. The enzyme activity was almost completely lost at 60°C and at pH4. It was slightly inhibited by Cu2+and Mn2+, and strongly by diisopropylfluorophosphate, phenylmethylsulfonylfluoride and chymostatin, suggesting that the enzyme may be a chymotrypsin-like protease..
44. Qing You Xia, Hiroshi Fujii, Takahiro Kusakabe, Yutaka Banno, Identification of three annexin ix isoforms generated by alternative splicing of the carboxyl-terminal exon in silkworm, bombyx mori, Insect Biochemistry and Molecular Biology, 10.1016/S0965-1748(01)00074-1, 32, 1, 9-14, 2001.01, Annexins (ANXs) are a family of structurally related proteins with Ca2--dependent phospholipid- binding properties. Here we report the cloning of three cDNAs each encoding annexin IX (ANX IX) isoforms from unfertilized eggs of the silkworm, Bombyx mori. The analysis of exon/intron structures showed that the three mRNAs, named ANX IX-A (2300 bp), ANX IX-B (1884 bp) and ANX IX-C (1409 bp), respectively, were generated from a single gene by alternative usage of a 3′-splice site of the last exon. Thus the three isoforms have an identical sequence from amino acid residues 1 to 307 and this region shows approximately 77% identity to Drosophila melanogaster ANX IX. Only amino acid residues 308-324 (A) or 308-323 (B and C), which correspond to the C-terminal tail, are different in the three proteins. A RT-PCR analysis indicated that the three isoforms of silkworm ANX IX were specifically expressed in various larval tissues and development stages. Interestingly, the C-terminal tail in ANXs I, II and V were previously confirmed as a binding region for protein kinase C. Thus generation of the three ANX IX isoforms in the silkworm, that are different from other ANXs, may have a functional significance other than binding to Ca2+..
45. Bannno, Y., Fujii, H. and Doira, H., Inheritance of one cocoon shape mutant named Flossycocoon of the silkworm, Bombyx mori., Journal of insect biotechnology and sericology, 70, 189-192, 2001.01.
46. Yukuhiro, K., Sezutsu, H., Itoh, M., Shimizu, K., and Banno, Y., Significant Levels of Sepuence Divergence and Gene Rearrangements have Occurred Between the MitochondrialGenomes of the Wild Mulberry Silkmoth, Bombyx mandarina, and its Close Relative, the Domesticated Silkmoth, Bombyx mori., Mol. Biol. Evol., 19, 8, 1385-1389, 19(8), 1385-1389, 2002.01.
47. Minoru Ujita, Akihiro Kimura, Daisuke Nishino, Eiji Yokoyama, Yutaka Banno, Hiroshi Fujii, and Akira Hara., Specific Binding of Silkworm Bombyx mori 30-kDa Lipoproteins to Carbohydrates Containing Glucose, Biosci. Biotechnol. Biochem., 66, 10, 2264-2266, 66(10), 2264-2266, 2002.01.
48. Tabunoki, H., Tanaka, H., Fujii, H., Banno, Y., Jouni, Z., Kobayashi, M., Sato, R., Maekawa, H.,Tsuchida, K., Isolation, characterization, and cDNA sequence of a carotenoid-binding protein from the silk gland of Bombyx mori., J. Biol. Chem., 10.1074/jbc.M204507200, 277, 35, 32133-32140, 277(35), 32133-32140, 2002.01.
49. Ningjia He, Masatoshi Yakiyama, Hiroshi Fujii, Yutaka Banno, Kohji Yamamoto., Genomic structure and expression analysis of the gene encoding a silkworm basic Kunitz-type chymotrypsin inhibitor, Biochimica et Biophysica Acta, 10.1016/S0167-4781(03)00118-0, 1628, 1, 71-77, 1628(2003) 71-77, 2003.01.
50. N. Komoto, H. Sezutsu, K. Yukuhiro, Y. Banno. H. Fujii., Mutations of the silkworm molybdenum cofactor sulfurase gene, og, cause translucent larval skin, Insect Biochemistry and Molecular Biology, 10.1016/S0965-1748(03)00006-7, 33, 4, 417-427, 33,417-427, 2003.01.
51. Kohji Yamamoto, Hiroshi Fujii, Yutaka Banno, Yoichi Aso and Masatsune Ishiguro., Polymorphism of Prophenoloxidase in the Silkworm, Bombyx mori, J. Fac. Agr., Kyushu Univ., 47, 2, 319-324, 47(2), 319-324, 2003.01.
52. Song Fangzhou, Zhang Pingbo, Yi Faping, Hong Xijun, Lu Cheng, Yutaka Banno, Hiroshi Fujii & Katsumi Koga., Study on fibroin heavy chain of the silkworm Bombyx mori by fluorescence in situ hybridization(FISH), SCIENCE IN CHINA(Series C), 10.1007/BF02879755, 45, 6, 663-+, vol.45 No.6, 2003.01.
53. Nakajima, Y., Nakamura, T., Banno, Y., Fujimoto, H., Hashido, K., Shiino, T., Tsuchida, K., Takada, N., and Maekawa, H., Comparison of mariner-like elements among Bombyx mandarina individuals inhabiting East Asia in the light of the segregation of B. mori and B. madarina genomes, International J. of Wild Silkworm and Silk, 8, 57-64, 2003.01.
54. Banno Y., Nakamura T., Nagashima E., Fujii H., Doira H., M chromosome of the wild silkworm, Bombyx mandarina(n=27), corresponds to two chromosomes in the domesticated silkworm, Bombyx mori(n=28)., Genome, 10.1139/G03-112, 47, 1, 96-101, 47, 96-101, 2004.01.
55. Takahiro Kusakabe, K. Kido, K. Kita, Yutaka Banno, H. Mon, Y. Kawaguchi, K. Koga, Analysis of artificial and spontaneous parthenogenetic development in mosaic mutations and the parthenogenetic strain of bombyx mori, Invertebrate Reproduction and Development, 10.1080/07924259.2004.9652579, 45, 2, 101-108, 2004.01, Interesting phenotypes found in male-female mosaic mutations of the silkworm are thought to result from the abnormal activation of a polar body due to the defect on the inactivation system. In addition to these mosaic mutations, a parthenogenetic strain (m90) of silkworm has been identified. The precise mechanism of parthenogenetic development in this strain is unknown, but it is possible that a defect of the polar body inactivation system plays a crucial role here, too. To understand the molecular mechanisms of polar body inactivation, we investigated the process of artificial and spontaneous parthenogenetic development using the mosaic mutations and the m90 strain. Both the pigmentation and the increase in DNA content were used to monitor development. In order to determine whether parthenogenesis in these strains is caused by incomplete meiotic division or fertilization of a polar body with the egg nucleus, the chromosome compositions were analyzed using an insertion sequence in the testis-specific tektin gene as a molecular marker. It was confirmed that parthenogenesis in the mosaic mutations was partly caused by the fertilization of a polar body nucleus with the egg nucleus. In contrast, parthenogenetic development in the m90 strain is apparently caused by incomplete meiotic division..
56. Tsuchida K., Jouni Z.E., Gardetto J., Kobayashi Y., Tabunoki H., Azuma M., Sugiyama H., Takada N., Maekawa H., Banno Y., Fujii H., Iwano H.,Wells M., Characterization of the carotenoid-binding protein of the Y-gene dominant mutants of Bombyx mori., Journal of Insect Physiology, 10.1016/j.jinphys.2004.02.006, 50, 4, 363-372, 50(4), 363-372, 2004.01.
57. Tabunoki H., Higurashi S., Ninagi O., Fujii H., Banno Y., Nozaki M., Kitajima M.,Miura N., Atsumi S., Tsuchida K., Maekawa H., Sato R., A carotenoid- binding protein (CBP) plays a crucial role in cocoon pigmentation of silkworm (Bombyx mori) larvae., FEBS Letters, 10.1016/j.febslet.2004.04.067, 567, 2-3, 175-178, 567, 175-178, 2004.01.
58. Tsuchida K., Katagiri C., Tanaka Y., Tabunoki H., Sato R., Maekawa H., Takada N.Banno Y., Fujii H. and Jouni Z.E., The Basis for Colorless Hemolymph and Cocoons in the Y-gene recessive Bombyx. mori Mutants: A Defect in the Cellular Uptake of Carotenoids., Journal of Insect Physiology, in press, 10.1016/j.jinsphys.2004.08.001, 50, 10, 975-983, 2004.01.
59. Ninjia He, Hiroshi Fujii, Takahiro Kusakabe, Yoichi Aso, Yutaka Banno, Kohji Yamamoto, Overexpression in Escherichia coli and purification of recombinant CI-b1, a Kunitz-type chymotrypsin inhibitor of silkworm., Protein Expr. Purif., 10.1016/j.pep.2004.07.010, 38, 1, 9-16, 38(1), 9-16, 2004.01.
60. Pingbo Zhang, Kohji Yamamoto, Yongqiang Wang, Yutaka Banno, Hiroshi Fujii, Fumio Miake, Nobuhiro Kashige, and Yoichi Aso, Utility of dry gel from two-dimensional electrophoresis for peptide mass fingerprinting analysis of silkworm proteins., Biosci. Biotechnol. Biochem., 68 (10), 2148-2154, 2004.01.
61. Wang yongqiang, Zhang Pingbo, Hiroshi Fujii, Yutaka Banno, Kohji Yamamoto,Yoichi Aso, Proteomis studies of lipopolysacccharide-induced polypeptides in the silkworm, Bombyx mori, Biosci. Biotechnol. Biochem., 10.1271/bbb.68.1821, 68, 8, 1821-1823, 68 (8), 1821-1823, 2004.01.
62. Qiaoling Zhao, Ninjia He, Koji Shirai, Hiroshi Fujii, Yutaka Banno, and Kohji Yamamoto, Purification and characterization of chymotrypsin inhibitor CI-3 from hemolymph of silkworm, Bombyx mori., J. Fac. Agr., Kyushu Univ., 49, 1, 93-99, 49 (1), 93-99, 2004.01.
63. Ningjia He, Yoichi Aso, Hiroshi Fujii, Yutaka Banno, and Kohji Yamamoto, In vivo and In vitro Interactions of the Bombyx mori Chymotrypsin Inhibitor b1 with Escherichia coli., Biosci. Biotechnol. Biochem., 10.1271/bbb.68.835, 68, 4, 835-840, 68 (4), 835-840, 2004.01.
64. Yutaka Banno, Kazuhiro Nishikawa, Akihiro Nishisaka, Kei Tamura, Kohji Yamamoto, Hiroshi Fujii, Inheritance of a body shape mutant named "Short Segment" of the domesticated silkworm, Bombyx mori, Journal of Insect Biotechnology and Sericology, 73, 1, 35-37, 2004.02, A dominant mutant, "Short Segment" (symbol Seg), was discovered as a spontaneous occurrence. Homozygotes and heterozygotes for Seg possess short body in length in each segment during all the larval stages. Linkage analysis showed that Seg was linked with odk and Nl, which had been mapped at the 32.5 and 35.2 positions, respectively, on the 14th linkage group. The Seg gene was precisely localized by a three-point experiment involving odk, Nl, and Seg. The recombination value between odk and Seg was calculated to be 0.60%, between Seg and NI 2.51% and between odk and Nl, 3.11%. Hence, the arrangement of these three gene loci on the chromosome is in the order odk-Seg-Nl. Taking a correction factor into account, the locus of Seg was determined to be 33.0 of the 14th linkage group..
65. Kohji Yamamoto, Stacey A. Rutherford, Malini Rajagopalan, Hideki Hirakawa, Satoru Kuhara, Yutaka Banno, Hiroshi Fujii, Murty V.V.S. Madiraju, Heterogeneity of dnaB Locus of Mycobacterium avium-intracellulare Complex, Journal of the Faculty of Agriculture, Kyushu University, 49, 2, 375-381, 2004.10, Analysis of the dnaB gene, homologue of the Escherichia coli replicative DNA helicase DnaB, from various Mycobacterium intracellulare complex strains revealed their dnaB genes were heterogeneity. We found that intein was included in-frame in the dnaB locus of M. intracellulare and that the intein is highly similar to M. avium intein. Phylogenetic study showed intein sequences were remote from their own host, dnaB sequences and suggested that the horizontal transfer had occurred among Mycobacterium avium-intracellulare complex strains..
66. Yamamoto K., Zhang P., He N., Wang Y., Aso Y., Banno Y., Fujii H., Molecular and biochemical characterization of manganese-containing superoxide dismutase from the silkworm, Bombyx mori, Comparative Biochemistry & Physiology, 10.1016/j.cbpb.2005.09.002, 142, 4, 403-409, 142 (B) 403-409, 2005.01.
67. Yamamoto K., Zhang P., Miake F., Kashige N., Aso Y., Banno Y., Fujii H., Cloning, expression and characterization of theta-class glutathione S-transferase from the silkworm, Bombyx mori., Comparative Biochemistry & Physiology, 10.1016/j.cbpc.2005.04.012, 141, 3, 340-346, 141 (B) 340-346, 2005.01.
68. Yamamoto K., Banno Y., Fujii H., Miake F., Kashige N., Aso Y., Catalase from the silkworm, Bombyx mori: Gene Sequence, Distribution, and Overexpression., Insect Biochemistry and Molecular Biology, 10.1016/j.ibmb.2005.01.001, 35, 4, 277-283, 35 277-283, 2005.01.
69. Yamamoto K., Zhang P., Banno Y., Fujii H., Miake F., Kashige N., Aso Y., Superoxide Dismutase from the Silkworm, Bombyx mori: Sequence, Distribution, and Overexpression., Biosci. Biotech. Biochem., 10.1271/bbb.69.507, 69, 3, 507-514, 69 507-514, 2005.01.
70. Hiroaki Abe, Motoaki Seki, Fumi Ohbayashi, Nobuhiko Tanaka, Jun-ichi Yamashita, Tsuguru Fujii, Takeshi Yokoyama, Michiyoshi Takahashi, Yutaka Banno, Ken Sahara, Atsuo Yoshido, Jun-ichiro Ihara, Yuji Yasukochi, Kazuei Mita, Masahiro Ajimura, Masataka G Su, Partial deletions of the W chromosome due to reciprocal translocation in the silkworm Bombyx mori., Insect Mol Biol., 10.1111/j.1365-2583.2005.00565.x, 14, 4, 339-352, 14, 339-352., 2005.01.
71. Minoru Ujita, Yosuke Katsuno, Ichiro Kawachi, Yoshinori Ueno, Yutaka Banno, Hiroshi Fujii, Akira Hara, Glucan-binding activity of silkworm 30-kDa apolipoprotein and its involvement in defense against fungal infection, Bioscience, Biotechnology and Biochemistry, 10.1271/bbb.69.1178, 69, 6, 1178-1185, 2005.06, The silkworm Bombyx mori 30-kDa lipoproteins (6G1 and 19G1), major components of the hemolymph, were shown to bind to glucans. 6G1 apolipoprotein was expressed as a fusion protein with glutathione S-transferase in Escherichia coli and assayed for its binding activity. The purified recombinant 6G1 apolipoprotein specifically bound to β-glucan, but not to chitin, mannan, peptidoglycan, or oligosaccharide chains on glycoproteins. The β-glucan binding of the recombinant 6G1 was inhibited by laminaribiose and laminarin, a soluble glucan, but not by lipopolysaccharide or insect blood sugar, trehalose at physiological concentration. Furthermore, the recombinant 6G1 was shown to participate in the activation of prophenoloxidase cascade and to interfere with hyphal growth of the entomopathogenic fungus Paecilomyces tenuipes, injected into pupae of B. mori. These results suggest that 6G1 lipoprotein plays a role in the protection of B. mori against invading fungi..
72. Yuji Yasukochi, Yutaka Banno, Kohji Yamamoto, Marian R. Goldsmith, Hiroshi Fujii, Integration of molecular and classical linkage groups of the silkworm, Bombyx mori (n = 28), Genome, 10.1139/G05-023, 48, 4, 626-629, 2005.08, Previously published linkage groups (LGs) composed of molecular markers were assigned to classical LGs in the silkworm, Bombyx mori (n = 28). Four markers from the classical linkage map, og, w-1, Lp, and Pfl, were assigned to the molecular linkage maps using sequence tagged sites. In addition, linkage analysis was carried out using BF1 progeny between wild-type and mutant stocks carrying morphological phenotypic markers. As a result, the counterparts for 26 of 28 molecular LGs were identified with their counterparts of the classical LGs. Two visible markers, Sel and Xan, representing different classical LGs, were found to be linked..
73. Pingbo Zhang, Kohji Yamamoto, Yoichi Aso, Yutaka Banno, Daisuke Sakano, Yongqiang Wang, Hiroshi Fujii, Proteomic studies of isoforms of the P25 component of Bombyx mori fibroin, Bioscience, Biotechnology and Biochemistry, 10.1271/bbb.69.2086, 69, 11, 2086-2093, 2005.12, It is recognized that P25 is one of three polypeptide components of the fibroin synthesized in the larval silk gland (SG) of silkworm, having two glycosylated isoforms. In the present study, however, eight P25 isoforms were separated by proteomics, including two-dimensional gel electrophoresis of whole SG proteins, and were identified by the peptide mass fingerprinting method. Four of the eight isoforms were identified as Bombyx mandarina P25s, although the SG of Bombyx mori has never been considered to contain the P25 from B. mandarina. It is suggested that this diversity of P25 isoforms depends on phosphorylation modification in addition to glycosylation..
74. Fujii T., Tanaka N., Yokoyama T., Ninaki O., Oshiki T., Ohnuma A., Tazima Y., Banno Y., Ajimura M., Mita K., Seki M., Ohbayashi F., Shimada T., Abe H., The female-killing chromosome of the silkworm, Bombyx mori, was generated by translocation between the Z and W chromosomes, Genetica, 127:253-265, 2006.01.
75. Kimiko Yamamoto, Junko Narukawa, Keiko Kadono-Okuda, Junko Nohata, Motoe Sasanuma, Yoshitaka Suetsugu, Yutaka Banno, Hiroshi Fujii, Marian R. Goldsmith, Kazuei Mita, Construction of a Single Nucleotide Polymorphism Linkage Map for the Silkworm, Bombyx mori, Based on Bacterial Artificial Chromosome End Sequences, Genetics Society of America, 173:151-161, 2006.05.
76. Pingbo Zhang, Yoichi Aso, Kohji Yamamoto, Yutaka Banno, Yongqiang Wang, Kozo Tsuchida, Yutaka Kawaguchi, Hiroshi Fujii, Proteome analysis of silk gland proteins from the silkworm, Bombyx mori, Proteomics, 2586-2599, 2006.06.
77. Yoshinori Ueno, Masao Takao, Ningjia He, Kohji Yamamoto, Yutaka Banno, Hiroshi Fujii, Genetic analysis of basic chymotrypsin inhibitors in the hemolymph of the silkworm, Bombyx mori, Journal of Insect Biotechnology and Sericology, 75, 2, 65-69, 2006.06, Using an acidic polyacrylamide gel electrophoresis followed by activity staining, three basic chymotrypsin inhibitors have been found in larval hemolymph of the silkworm, Bombyx mori. These components, named Cls-b1, b2 and b3 in order of migration towards the cathode, were subjected to linkage analysis. Results indicated that the genes coding for Cls-b1 and b2 were found to be novel multiple alleles of the Ict-H locus linked with the Y gene on the second linkage group..
78. Yoshinori Ueno, Ningjia He, Minoru Ujita, Kohji Yamamoto, Yutaka Banno, Hiroshi Fujii, Yoichi Aso, Silkworm midgut proteins interacting with a hemolymph protease inhibitor, CI-8, Bioscience, Biotechnology and Biochemistry, 10.1271/bbb.50548, 70, 7, 1557-1563, 2006.08, CI-8 is the chymotrypsin inhibitor in hemolymph from the silkworm, Bombyx mori. It occurs in the midgut at the spinning stage of larva, but little information on the mechanism of its uptake in the midgut is available. We found that two polypeptides interacting with CI-8 are in the midgut membrane, and we purified them using a biotinylated CI-8, viz., p29 and p60, having molecular sizes of 29 kDa and 60 kDa respectively. The structures of p29 and p60 were examined by N-terminal amino acid sequencing and peptide mass mapping, including tryptic digestion. p29 was highly similar to the matured 19G1-30K lipoprotein from hemolymph, but p60 was novel. Purified p29 was recognized by anti-19G1-30K antibody, and was confirmed to be similar to 19G1-30K. The antibody also neutralized the CI-8 binding ability of p29 in the midgut membrane. p29 and p60 are perhaps proteinaceous factors involved in the uptake of CI-8 into the midgut through the membrane..
79. Naoko Takada, Emiko Yamauchi, Hirofumi Fujimoto, Yutaka Banno, Kozo Tsuchida, Kazuo Hashido, Yumiko Nakajima, Zhenli Tu, Masateru Takahashi, Hiroshi Fujii, Hajime Fugo, Hideaki Maekawa, A novel indicator for radiation sensitivity using the wing size reduction of Bombyx mori pupae caused by γ-ray irradiation, Journal of Insect Biotechnology and Sericology, 75, 3, 161-165, 2006.10, The wings of Bombyx mori are known to become smaller when irradiated with γ-rays at the larval stage. This was considered to be a non-stochastic effect wherein the wing-size reduction curve, plotted vs. the irradiation dose, shows a threshold. Here we propose a new indicator for radiation sensitivity, named the wingless dose 50 (WLD50), which was obtained from the inflection point of wing-size data normalized by the body-size changes and plotted against the irradiation dose. This indicator was confirmed to serve as a tool to compare irradiation sensitivity among B. mori strains..
80. Yan Meng, Chaoliang Liu, Kunihiro Shiomi, Masao Nakagaki, Yutaka Banno, Zenta Kajiura, Genetic variations in the vitellogenin of Japanese populations of the wild silkworm Bombyx mandarina, Journal of Insect Biotechnology and Sericology, 75, 3, 127-134, 2006.10, We cloned and sequenced cDNA of Bombyx mandarina vitellogenin (BmaVg) inhabiting Japan. The complete sequence of BmaVg cDNA without a poly A tall is 5733 bp long and encodes 1780 amino acid residues among which are typical highly conserved sequences and common features found in most insects. Multiple alignment of vitellogenin sequences of Japanese Bombyx mandarina, the Chinese one, and Bombyx mori showed that the nucleotide sequence of Japanese BmaVg cDNA shared 99.5% identity with Chinese BmaVg and 98.2% identity with Bombyx mori vitellogenin (BmoVg), and that the amino acid sequence of Japanese BmaVg cDNA shared 99.6% homology with Chinese BmaVg and 97.5% homology with BmoVg. These findings indicate that BmaVg (JPN) is more similar to BmaVg (CHN) than to BmoVg, which was also confirmed through a comparison of the mutation frequencies of both nucleotide and amino acid sequences. The phylogenetic tree of Vgs demonstrates that BmaVgs of individuals collected from eighteen prefectures in Japan primarily cluster in a group with BmaVg (CHN) and without BmoVg, suggesting that the sequence variation between the two BmaVgs arose after divergence of B. mori from B. mandarina..
81. Kohji Yamamoto, P. B. Zhang, Yutaka Banno, H. Fujii, Identification of a sigma-class glutathione-S-transferase from the silkworm, Bombyx mori, Journal of Applied Entomology, 10.1111/j.1439-0418.2006.01092.x, 130, 9-10, 515-522, 2006.12, A cDNA that encodes a glutathione-S-transferase belonging to sigma class (GSTS1) from the silkworm, Bombyx mori was cloned by reverse transcriptase-polymerase chain reaction and sequenced. The deduced amino acid sequence revealed 66%, 48% and 41% identity to sigma-class GSTs from Manduca sexta, Blattella germanica and Anopheles gambiae, respectively. The GSTS1 was also estimated to be close to those GSTs in a phylogenetic tree. GSTS1 mRNA was widely distributed in various tissues. The recombinant GST (rGSTS1) was functionally overexpressed in Escherichia coli in a soluble form, purified to homogeneity and characterized. The pH-optimum of rGSTS1 was around pH 8. The rGSTS1 retained more than 75% of its original activity after incubation at pH 4-9. Incubation for 30 min at temperatures below 40°C did not affect its activity. rGSTS1 was able to catalyse the reaction of glutathione with 1-chloro-2,4-dinitrobenzene, the universal substrate for GST, as well as with 4-hydroxynonenal, the product of lipid peroxidation..
82. Kohji Yamamoto, Pingbo Zhang, Yutaka Banno, Fumio Miake, Kazuhiro Nishikawa, Akifumi Nishisaka, Kei Tamura , Comparative Analysis of Cuticle Proteins of the Silkworm by Two-Dimensional Electrophoresis , Sericologia , 47, 1, 53-58, 2007.01.
83. Kohji Yamamoto, Taro Okada, Yutaka Banno, Hiroshi Fujii, Expression of Messenger RNA Encoding a Y-Box Protein of the Silkworm, Bombyx mori and Accumulation in Ovarian Follicle Cells, Sericologia, Vol.47, No.1, pp.41-48, 2007.01.
84. Pingbo Zhang, Yoichi Aso, Hiroyuki Jikuya, Takahiro Kusakabe, Jae Man Lee, Yutaka Kawaguchi, Kohji Yamamoto, Yutaka Banno, Hiroshi Fujii, Proteomic Profiling of the Silkworm Skeletal Muscle Proteins during Larval-Pupal Metamorphosis, Journal of Proteome Research, Vol.6, No.6, pp.2295-2303, 2007.02.
85. Kohji Yamamoto, Hiroshi Fujii, Yoichi Aso, Yutaka Banno, Katsumi Koga, Expression and characterization of a sigma-class glutathione S-transferase of the fall webworm, Hyphantria cunea, Bioscience, Biotechnology and Biochemistry, 10.1271/bbb.60592, 71, 2, 553-560, 2007.03, A cDNA encoding glutathione S-transferase (GST) of the fall webworm, Hyphantria cunea, was cloned by reverse transcriptase-polymerase chain reaction. The resulting clone (hcGST) was sequenced and deduced for amino acid sequence, which revealed 87, 59, and 42% identities to Sigma-class GSTs from Bombyx mori, Manduca sexta, and Blattella germanica respectively. A recombinant hcGST protein (rhcGST) was functionally overexpressed in Escherichia coli cells in a soluble form and purified to homogeneity. rhcGST retained more than 75% of its original GST activity after incubation at pHs 6 to 11. Incubation for 30 min at temperatures below 50°C scarcely affected the activity. rhcGST was able to catalyze the reaction of glutathione with 1- chloro-2,4-dinitrobenzene, a universal substrate for GST, as well as with 4-hydroxynonenal, a product of lipid peroxidation. We also found that as compared to B. mori Sigma-class GST, rhcGST had a higher affinity for fenitrothion, an organophosphorus insecticide..
86. Takashi Sakudoh, Hideki Sezutsu, Takeharu Nakashima, Isao Kobayashi, Hirofumi Fujimoto, Keiro Uchino, Yutaka Banno, Hidetoshi Iwano, Hideaki Maekawa, Toshiki Tamura, Hiroshi Kataoka, Kozo Tsuchida, Carotenoid silk coloration is controlled by a carotenoid-binding protein, a product of the Yellow blood gene, Proceedings of the National Academy of Sciences of the United States of America, 10.1073/pnas.0702860104, 104, 21, 8941-8946, 2007.05, Mechanisms for the uptake and transport of carotenoids, essential nutrients for humans, are not well understood in any animal system. The Y (Yellow blood) gene, a critical cocoon color determinant in the silkworm Bombyx mori, controls the uptake of carotenoids into the intestinal mucosa and the silk gland. Here we provide evidence that the Y gene corresponds to the intracellular carotenoid-binding protein (CBP) gene. In the Y recessive strain, the absence of an exon, likely due to an incorrect mRNA splicing caused by a transposon-associated genomic deletion, generates a nonfunctional CBP mRNA, resulting in colorless hemolymph and white cocoons. Enhancement of carotenoid uptake and coloration of the white cocoon was achieved by germ-line transformation with the CBP gene. This study demonstrates the existence of a genetically facilitated intracellular process beyond passive diffusion for carotenoid uptake in the animal phyla, and paves the way for modulating silk color and lipid content through genetic engineering..
87. Yutaka Banno, Kazuhiro Nishikawa, Akifumi Nishisaka, Kei Tamura, Kohji Yamamoto, Hiroshi Fujii, Erratum
"Inheritance of a body shape mutant named "short segment" of the domesticated silkworm, Bombyx mori" (Journal of Insect Biotechnology and Sericology vol. 73 (1) (35-37)), Journal of Insect Biotechnology and Sericology, 76, 2, 2007.06.
88. Man Lee, Hiroaki Mon, Masateru Takahashi, Naoya Kawakami, Hitoshi Mitsunobu, Yutaka Banno, Katsumi Koga, Keiro Uchino, Yutaka Kawaguchi, Takahiro Kusakabe, Screening of high-permissive silkworm strains for efficient recombinant protein production in Autographa californica nuclear polyhedrosis virus (AcNPV), Journal of Insect Biotechnology and Sericology, 76, 2, 101-105, 2007.06, The baculovirus expression system (BES) using Autographa california nuclear polyhedrosis virus (AcNPV) has been extensively utilized for the high-level expression of sufficient quantities of recombinant proteins in a broad taxonomic range of insect cell lines and insect larvae. In the case of the silkworm, Bombyx mori, however, the BES using AcNPV tends to be much less exploited because AcNPV is believed to be inefficient in the replication of B. mori and cell lines derived from it. In this study, we have searched for high-permissive silkworm strains with higher production levels of a recombinant protein, by screening the susceptibility of 163 silkworm strains to the recombinant AcNPV, which expresses firefly luciferase. Based on their relative luciferase expression levels in larval hemocytes, the silkworm strains tested were divided into 3 groups: 5 high-permissive strains, 74 middle-permissive strains, and 84 low-permissive strains. Among the 5 high-permissive strains (c11, d17, f10, f38, and 1312), d17 was the most productive, showing the luciferase activity of 3.5 × 104 ± 1,764 relative light units (RLU) per microgram of cell proteins. This remarkable susceptibility indicated that the silkworm d17 strain is very useful for large-scale protein production by the BES using AcNPV..
89. Yuichi Kawanishi, Reiko Takaishi, Yutaka Banno, Hirofumi Fujimoto, Si Kab Nho, Hideaki Maekawa, Yumiko Nakajima, Sequence comparison of mariner-like elements among the populations of Bombyx mandarina inhabiting China, Korea and Japan, Journal of Insect Biotechnology and Sericology, 76, 2, 79-87, 2007.06, Mariner-like elements (MLEs) were isolated from Chinese, Korean and Japanese individuals of Bombyx mandarina by the long PCR methods using the inverted terminal repeat of the Hyalophora cecropia MLE as a primer. The amplified clones of about 1.3 kbp in size were regarded to be full-length MLEs and termed Cecropia-ITR-MLEs. The sequences of the elements were determined, trimmed to eliminate any insertions and used to construct a phylogenetic tree. As a result, the Cecropia-ITR-MLE sequences of B. mandarina could be divided into major groups here called the Continental type and the Japanese type. Chinese and Korean specimens mostly belonged to the former group, which is very diverse and may well consist of subgroups. A majority of Honshu (the main island of Japan) specimens belonged to the latter, which is a much more closely related group of sequences. Interestingly, Tsushima (south of Korea) and Hokkaido (close to Russia) specimens mainly fell into the former type, whereas Fukuoka (south of Tsushima) individuals included both..
90. Ping Zhao, Qingyou Xia, Juan Li, Hiroshi Fujii, Yutaka Banno, Zhonghuai Xiang, Purification, characterization and cloning of a chymotrypsin inhibitor (CI-9) from the hemolymph of the silkworm, Bombyx mori, Protein Journal, 10.1007/s10930-007-9077-0, 26, 5, 349-357, 2007.08, Hemolymph chymotrypsin inhibitor 9 (CI-9) from the hemolymph of the silkworm, Bombyx mori, was purified by ammonium sulfate precipitation, Butyl Toyopearl hydrophobic chromatography, gel filtration through Sephadex C-50 and chymotrypsin-sepharose 4B affinity chromatography. Checked by Native PAGE and SDS-PAGE in combination with silver staining, the final preparation appeared homogeneous. In tricine SDS-PAGE, CI-9 displayed a molecular weight of 7.5 kD, which was determined to be 7167 Da with the Voyager TOFMass analyser. The pI value for CI-9, revealed by 2D-PAGE (two-dimensional polyacrylamide gel electrophoresis), was 4.3. CI-9 exhibited inhibitory activity at a temperature as high as 100°C and a stability against a wide range of pH (1-12). In N-terminal amino-acid analysis of CI-9, 40 amino acid residues were obtained. The C-terminal 22 amino acid residues were deduced by subsequently cloned cDNA and genomic fragments. MW and pI of CI-9 were predicted to be 7170.98 Da and 4.61, respectively, on the website. Its low molecular weight, high stability, conserved active site and Kunitz domain showed that CI-9 is a Kunitz-type CI. The difference of sequence and pI between CI-9 and other Kunitz type CIs indicated that it is a novel chymotrypsin inhibitor..
91. Man Lee, Hiroaki Mon, Chisa Yasunaga-Aoki, Masateru Takahashi, Naoya Kawakami, Hitoshi Mitsunobu, Yutaka Banno, Katsumi Koga, Keiro Uchino, Yutaka Kawaguchi, Takahiro Kusakabe, Erratum
"Screening of high-permissive silkworm strains for efficient recombinant protein production in Autographa californica nuclear polyhedrosis virus (AcNPV)" (Journal of Insect Biotechnology and Sericology vol. 76 (2) (101-105)), Journal of Insect Biotechnology and Sericology, 76, 3, 169, 2007.10.
92. Wajiro Hara, Stephen Sosnicki, Yutaka Banno, Hirofumi Fujimoto, Naoko Takada, Hideaki Maekawa, Hiroshi Fujii, Michael A. Wells, Kozo Tsuchida, Mapping analysis of carotenoids-binding protein of Bombyx mori by restriction fragment length polymorphism, Journal of Insect Biotechnology and Sericology, 76, 3, 149-154, 2007.10, The gene for carotenoid binding protein (CBP) in the silkworm, Bombyx mori, was mapped. The linkage group for the CBP gene was identified by scanning linkage analysis of the restriction fragment length polymorphism (RFLP) pattern, and its genetic mapping with respect to known molecular markers within the group was established. The CBP gene was found to be linked to RFLP linkage group 19 which is equivalent to the phenotypic linkage group 2. The linkage map at 55 cM in length was constructed for this linkage group, with CBP mapped at 21 cM in length. When compared to a phenotypic map of chromosome 2, this result suggests that the gene that encodes CBP is the Y-gene..
93. Kohji Yamamoto, Makoto Kimura, Yoichi Aso, Yutaka Banno, Katsumi Koga, Characterization of superoxide dismutase from the fall webworm, Hyphantria cunea
Comparison of its properties to superoxide dismutase from the silkworm Bombyx mori, Applied Entomology and Zoology, 10.1303/aez.2007.465, 42, 3, 465-472, 2007.12, Superoxide dismutase (SOD) is a regulatory enzyme involved in the degradation of superoxide anions in living organisms. In this study, we examined SOD from the fall webworm, Hyphantria cunea (hcSOD). A cDNA encoding hcSOD was cloned by reverse transcriptase-polymerase chain reaction. The deduced amino acid sequence of hcSOD revealed that six of seven highly conserved residues forming the Cu/Zn-binding sites were present. The hcSOD mRNA and the enzyme activity were distributed in larval tissues including fat body, midgut and hemocyte of H. cunea. A recombinant hcSOD (rhcSOD) functionally overexpressed in Escherichia coli in a soluble form that was purified to homogeneity. It was stable at pHs between 5 and 11. Incubation for 30 min at temperatures below 50°C scarcely affected the activity. All these properties, as well as CD spectra, of hcSOD or rhcSOD were highly similar to those of the Bombyx mori counterparts except for the number of Cu/Zn-binding sites and the specific activity dismutating superoxide anion to peroxide and oxygen..
94. Hiroshi Fujii, Yutaka Banno & Kohji Yamamoto, Polymorphic Variants of Superoxide Dismutase and Glucose 6-Phosphate Dehydrogenase in Hemolymph of the Silkworm, Bombyx Mori, Sericologia, Vol.48, No.2, pp.133-140, 2008.01.
95. Kimiko Yamamoto, Junko Nohata, Keiko Kadono-Okuda, Junko Narukawa, Motoe Sasanuma, Shun Ichi Sasanuma, Hiroshi Minami, Michihiko Shimomura, Yoshitaka Suetsugu, Yutaka Banno, Kazutoyo Osoegawa, Pieter J. de Jong, Marian R. Goldsmith, Kazuei Mita, A BAC-based integrated linkage map of the silkworm Bombyx mori, Genome Biology, 10.1186/gb-2008-9-1-r21, 9, 1, 2008.01, Background: In 2004, draft sequences of the model lepidopteran Bombyx mori were reported using whole-genome shotgun sequencing. Because of relatively shallow genome coverage, the silkworm genome remains fragmented, hampering annotation and comparative genome studies. For a more complete genome analysis, we developed extended scaffolds combining physical maps with improved genetic maps. Results: We mapped 1,755 single nucleotide polymorphism (SNP) markers from bacterial artificial chromosome (BAC) end sequences onto 28 linkage groups using a recombining male backcross population, yielding an average inter-SNP distance of 0.81 cM (about 270 kilobases). We constructed 6,221 contigs by fingerprinting clones from three BAC libraries digested with different restriction enzymes, and assigned a total of 724 single copy genes to them by BLAST (basic local alignment search tool) search of the BAC end sequences and high-density BAC filter hybridization using expressed sequence tags as probes. We assigned 964 additional expressed sequence tags to linkage groups by restriction fragment length polymorphism analysis of a nonrecombining female backcross population. Altogether, 361.1 megabases of BAC contigs and singletons were integrated with a map containing 1,688 independent genes. A test of synteny using Oxford grid analysis with more than 500 silkworm genes revealed six versus 20 silkworm linkage groups containing eight or more orthologs of Apis versus Tribolium, respectively. Conclusion: The integrated map contains approximately 10% of predicted silkworm genes and has an estimated 76% genome coverage by BACs. This provides a new resource for improved assembly of whole-genome shotgun data, gene annotation and positional cloning, and will serve as a platform for comparative genomics and gene discovery in Lepidoptera and other insects..
96. Yuichi Kawanishi, Reiko Takaishi, Miki Morimoto, Yutaka Banno, Si Kab Nho, Hideaki Maekawa, Yumiko Nakajima, A novel maT-type transposable element, BmamaT1, in Bombyx mandarina, homologous to the B. mori mariner-like element Bmmar6, Journal of Insect Biotechnology and Sericology, 77, 1, 45-52, 2008.02, A complex type marner-like element (MLE) was isolated by PCR with genomic DNA of Bombyx mandarina. The clones of this MLE, homologous to the 'Cecropia-ITR-MLE' in BmTNML locus that we have isolated previously from B. mori, were roughly classified into four groups according to the state of inserted elements: (1) Containing a retrotransposon (L1Bm) and another element, (2) containing L1Bm, alone, (3) containing another element alone and (4) without insertions. This inserted element was named BmamaT1, as it was homologous to the B. mori MLE called Bmmar6, which was previously reported to be similar to a maT-family member, Bmmar1. Many of the current Cecropia-ITR-MLE clones had, as the BmamaT1 insertion site, the sequence of TA/TATA, which may be a transposition target site. Moreover, the BmamaT1 elements that inserted into the Cecropia-ITR-MLE region were highly homogeneous, making one group when a phylogenetic tree was made together with BmamaT1 members isolated directly from the B. mandarina genome. Many of the inserted type BmamaT1 elements had complete ORF which: possessed the so-called DDD catalytic triad. All these findings were taken to indicate that a large fraction of the BmamaT1 elements are still highly active..
97. Fang Zhou Song, Ping An Chang, Ping Bo Zhang, Fa Ping Yi, Yong Ping Ma, Cheng Lu, Yutaka Banno, Hiroshi Fujii, Chromosomal localization of silkworm (Bombyx mori) sericin gene 1 and chymotrypsin inhibitor 13 using fluorescence in situ hybridization, Science in China, Series C: Life Sciences, 10.1007/s11427-008-0025-9, 51, 2, 133-139, 2008.02, The chromosomal locations of two single-copy genes, Ser-1 and Cl-13, in silkworm (Bombyx mori) were detected at the molecular cytogenetics level by fluorescence in situ hybridization in the study. The results showed that Ser-1 is located near the distal end of the 11th linkage group, relatively at the 12.5±1.4 position in pachytene; and that Cl-13 has been mapped near the distal end of the 2nd linkage group, relatively at the 8.2±1.2 position in pachytene. Furthermore, their location model map-FISH map on silkworm chromosome was drawn. The FISH technique and its application to silkworm are also discussed in this paper..
98. Yuichi Kawanishi, Yutaka Banno, Hirofumi Fujimoto, Si Kab Nho, Zhenli Tu, Kazuei Mita, Kozo Tsuchida, Naoko Takada, Hideaki Maekawa, Yumiko Nakajima, Method for rapid distinction of Bombyx mandarina (Japan) from B. mandarina (China) based on rDNA sequence differences, Journal of Insect Biotechnology and Sericology, 77, 2, 79-85, 2008.06, The EcoRI site is located between the 5.8S and 28S rRNA genes in Bombyx mandarina (Japan) (Japanese type rDNA) but is absent in the rDNA of B. mandarina (China)(B. mori type rDNA) including the domesticated B. mori. Digestion of PCR products amplified by primers of both flanking sides of this EcoRI site with EcoRI should only be successful for amplification products of B. mandarina (Japan) containing Japanese type rDNA. The distribution of Japanese type rDNA in Japan, Korea, and China was determined using this rapid detection method. Japanese type was not detected in China or in Korea despite having 27 chromosomes. These findings suggest that the choromosome fusion and the rDNA type separation were independent evolutionary events. Invasion of B. mori chromosome #11 into Japanese B. mandarina (Japan) was examined..
99. Kaoru Sato, Tomoko Matsuoka Matsunaga, Ryo Futahashi, Tetsuya Kojima, Kazuei Mita, Yutaka Banno, Haruhiko Fujiwara, Positional cloning of a bombyx wingless locus flügellos (fl) reveals a crucial role for fringe that is specific for wing morphogenesis, Genetics, 10.1534/genetics.107.082784, 179, 2, 875-885, 2008.06, Mutations at the flügellos (fl) locus in Bombyx mori produce wingless pupae and moths because of the repressed response of wing discs to ecdysteroid. Four recessive fl alleles occurred spontaneously and were mapped at 13.0 of the silkworm genetic linkage group 10. By positional cloning, we confirmed that the gene responsible for fl is fringe (fng) encoding Fng glycosyltransferase, which is involved in regulating the Notch signaling pathway. In four different fl alleles, we detected a large deletion of the fng gene in flk and nonsense mutations in fl, flo, and fln. In the wild-type (WT) silkworm, fng is expressed actively in the wing discs, brain, and reproductive organs from the fourth to final instars but barely in the other tissues tested. In situ hybridization showed that fng mRNA is expressed in the dorsal layer of the WT wing discs. The wingless (wg) mRNA, a downstream marker of Fng-mediated Notch signaling, is localized at the dorsoventral boundary in the WT wing discs but repressed markedly in the fl wing discs. Although null mutants of Drosophila fng result in postembryonic lethality, loss of fng function in Bombyx affects only wing morphogenesis, suggesting different essential roles for fng in tissue differentiation among insects..
100. Hiroko Tabunoki, Toru Shimada, Yutaka Banno, Ryoichi Sato, Hideyuki Kajiwara, Kazuei Mita, Jun ichi Satoh, Identification of Bombyx mori 14-3-3 orthologs and the interactor Hsp60, Neuroscience Research, 10.1016/j.neures.2008.03.007, 61, 3, 271-280, 2008.07, The 14-3-3 protein family consists of evolutionarily conserved, acidic 30 kDa proteins composed of seven isoforms named β, γ, ε, ζ, η, θ, and σ in mammalian cells. The dimeric complex of 14-3-3 isoforms, acting as a molecular adaptor, plays a central role in regulation of neuronal function. Since aberrant expression of 14-3-3 is identified in the brains of Alzheimer disease and Parkinson disease, a convenient insect model, if it is available, is highly valuable for studying a pathological role of 14-3-3 in neurodegeneration. Here, we identified the silkworm Bombyx mori 14-3-3 orthologs, ζ and ε isoforms highly homologous in amino acid sequences to the human 14-3-3ζ and 14-3-3ε. By Western blot, the expression of ζ and ε isoforms was identified at substantial levels in the first instar larva, markedly upregulated in the second instar larva, and the highest levels were maintained in the late stage of larva, the pupa, and the adult. Furthermore, by protein overlay and immunoprecipitation, we identified Hsp60 as a 14-3-3-binding partner. The 14-3-3 proteins interacted with the N-terminal fragment of Hsp60. The 14-3-3ζ and ε isoforms, along with Hsp60, were expressed widely with overlapping distribution in larval and adult tissues, including brain, fat body, silk gland, Malpighian tube, midgut, ovary, testis, antenna, and pheromone gland. These observations suggest that a molecular adaptor 14-3-3 and a molecular chaperone Hsp60 cooperate to achieve a wide range of cellular functions in B. mori..
101. H. Abe, T. Fujii, N. Tanaka, T. Yokoyama, H. Kakehashi, M. Ajimura, K. Mita, Yutaka Banno, Y. Yasukochi, T. Oshiki, M. Nenoi, T. Ishikawa, T. Shimada, Identification of the female-determining region of the W chromosome in Bombyx mori, Genetica, 10.1007/s10709-007-9210-1, 133, 3, 269-282, 2008.07, The W chromosome of the silkworm Bombyx mori is devoid of functional genes, except for the putative female-determining gene (Fem). To localize Fem, we investigated the presence of W-specific DNA markers on strains in which an autosomal fragment containing dominant marker genes was attached to the W chromosome. We produced new W-chromosomal fragments from the existing Zebra-W strain (T(W;3)Ze chromosome) by X-irradiation, and then carried out deletion mapping of these and sex-limited yellow cocoon strains (T(W;2)Y-Chu, -Abe and -Ban types) from different Japanese stock centers. Of 12 RAPD markers identified in the normal W chromosomes of most silkworm strains in Japan, the newly irradiated W(B-YL-YS)Ze chromosome contained three, the T(W;2)Y-Chu chromosome contained six, and the T(W;2)Y-Abe and -Ban chromosomes contained only one (W-Rikishi). To investigate the ability of the reduced W-chromosome translocation fragments to form heterochromatin bodies, which are found in nuclei of normal adult female sucking stomachs, we examined cells of the normal type p50 strain and the T(W;2)Y-Chu and -Abe strains. A single sex heterochromatin body was found in nuclei of p50 females, whereas we detected only small sex heterochromatin bodies in the T(W;2)Y-Chu strain and no sex heterochromatin body in the T(W;2)Y-Abe strain. Since adult females of all strains were normal and fertile, we conclude that only extremely limited region, containing the W-Rikishi RAPD sequence of the W chromosome, is required to determine femaleness. Based on a comparison of the normal W-chromosome and 7 translocation and W-deletion strains we present a map of Fem relative to the 12 W-specific RAPD markers..
102. Naoya Kawakami, Man Lee, Hiroaki Mon, Yuji Kubo, Yutaka Banno, Yutaka Kawaguchi, Katsumi Maenaka, Enoch Y. Park, Katsumi Koga, Takahiro Kusakabe, Efficient protein expression in Bombyx mori larvae of the strain d17 highly sensitive to B. mori nucleopolyhedrovirus, Molecular Biotechnology, 10.1007/s12033-008-9074-3, 40, 2, 180-185, 2008.10, The recombinant protein expression by Bombyx mori nucleopolyhedrovirus (BmNPV) infecting silkworm larvae or pupae may endow us with a potent system for the production of large eukaryotic proteins. However, the screening of silkworm strains ideally suited to this method has scarcely been conducted. In the present study, we injected recombinant BmNPV containing a reporter gene, luciferase or DsRed, into hemocoel of fifth instar larvae of selected 12 silkworm strains. Among them, the strain d17 is found to be the highest in reporter expression from the intrinsic polyhedrin promoter of Autographa californica NPV or the silkworm actin A3 promoter. These results suggest that the d17 strain is highly permissive for BmNPV replication and is the most likely candidate of a "factory" for large-scale expression using the BmNPV bacmid system..
103. Ryo Futahashi, Jotaro Sato, Yan Meng, Shun Okamoto, Takaaki Daimon, Kimiko Yamamoto, Yoshitaka Suetsugu, Junko Narukawa, Hirokazu Takahashi, Yutaka Banno, Susumu Katsuma, Toru Shimada, Kazuei Mita, Haruhiko Fujiwara, yellow and ebony are the responsible genes for the larval color mutants of the silkworm Bombyx mori, Genetics, 10.1534/genetics.108.096388, 180, 4, 1995-2005, 2008.12, Many larval color mutants have been obtained in the silkworm Bombyx mori. Mapping of melanin-synthesis genes on the Bombyx linkage map revealed that yellow and ebony genes were located near the chocolate (ch) and sooty (so) loci, respectively. In the ch mutants, body color of neonate larvae and the body markings of elder instar larvae are reddish brown instead of normal black. Mutations at the so locus produce smoky larvae and black pupae. F2 linkage analyses showed that sequence polymorphisms of yellow and ebony genes perfectly cosegregated with the ch and so mutant phenotypes, respectively. Both yellow and ebony were expressed in the epidermis during the molting period when cuticular pigmentation occurred. The spatial expression pattern of yellow transcripts coincided with the larval black markings. In the ch mutants, nonsense mutations of the yellow gene were detected, whereas large deletions of the ebony ORF were detected in the so mutants. These results indicate that yellow and ebony are the responsible genes for the ch and so loci, respectively. Our findings suggest that Yellow promotes melanization, whereas Ebony inhibits melanization in Lepidoptera and that melanin-synthesis enzymes play a critical role in the lepidopteran larval color pattern..
104. Yan Meng, Susumu Katsuma, Takaaki Daimon, Yutaka Banno, Keiro Uchino, Hideki Sezutsu, Toshiki Tamura, Kazuei Mita, Toru Shimada, The silkworm mutant lemon (lemon lethal) is a potential insect model for human sepiapterin reductase deficiency, Journal of Biological Chemistry, 10.1074/jbc.M900485200, 284, 17, 11698-11705, 2009.04, Tetrahydrobiopterin (BH4) is an essential cofactor for aromatic acid hydroxylases, which control the levels of monoamine neurotransmitters. BH4 deficiency has been associated with many neuropsychological disorders. An inherited defect in BH4 biosynthesis is caused by the deficiency of sepiapterin reductase (SPR), which catalyzes the biosynthesis of BH4 from guanosine triphosphate at the terminal step. The human SPR gene has been mapped at the PARK3 locus, which is related to the onset of Parkinson disease. In this study, we report that mutant strains, lemon (lem) and its lethal allele lemon lethal (leml) with yellow body coloration, of the silkworm Bombyx mori could be used as the first insect model for human SPR deficiency diseases. We demonstrated that mutations in the SPR gene (BmSpr) were responsible for the irregular body coloration of lem and leml. Moreover, biochemical analysis revealed that SPR activity in leml larvae was almost completely diminished, resulting in a lethal phenotype that the larvae cannot feed and that die immediately after the first ecdysis. Oral administration of BH4 and dopamine to leml larvae effectively increased their survival rates and feeding abilities. Our data demonstrate that BmSPR plays a crucial role in the generation of BH4, and monoamine neurotransmitters in silkworms and the lem (lem1) mutant strains will be an invaluable resource to address many questions regarding SPR and BH4 deficiencies..
105. Kohji Yamamoto, Sumiharu Nagaoka, Yutaka Banno, Yoichi Aso, Biochemical properties of an omega-class glutathione S-transferase of the silkmoth, Bombyx mori, Comparative biochemistry and physiology. Toxicology & pharmacology : CBP, 10.1016/j.cbpc.2008.10.108, 149, 4, 461-467, 2009.05, A cDNA encoding an omega-class glutathione S-transferase of the silkmoth, Bombyx mori (bmGSTO), was cloned by reverse transcriptase-polymerase chain reaction. The resulting clone was sequenced and deduced for amino acid sequence, which revealed 40, 40, and 39% identities to omega-class GSTs from human, pig, and mouse, respectively. A recombinant protein (rbmGSTO) was functionally overexpressed in Escherichia coli cells in a soluble form and purified to homogeneity. rbmGSTO was able to catalyze the biotranslation of glutathione with 1-chloro-2,4-dinitrobenzene, a model substrate for GST, as well as with 4-hydroxynonenal, a product of lipid peroxidation. This enzyme was shown to have high affinity for organophosphorus insecticide and was present abundantly in silkmoth strain exhibiting fenitrothion resistance. These results indicate that bmGSTO could be involved in the increase in level of insecticide resistance for lepidopteran insects..
106. Majibur Rahman Khan, Mikihiko Miura, Yutaka Banno, Hideaki Morikawa, Ying Chen, 3D analysis of the spinning behavior of flossy cocoon mutants in the silkworm, Bombyx mori, Journal of Insect Biotechnology and Sericology, 78, 3, 139-147, 2009.10, To analyze the spinning behavior of flossy cocoon mutants in the silkworm, Bombyx mori, the spinning characteristics of two mutant larvae: W07+, which produced spherical-shaped flossy cocoons, and WO7q constructed peanut-shaped tight cocoons were recorded on videotapes from two different angles and analyzed for spinning behavior by the three dimensional computer graphics software, 3DASBS. WO7+ silkworms constructed an appropriate shape of cocoon very rapidly. On the other hand, WO7q silkworms produced cocoon very slowly and needed more time to form a full cocoon shape than W07+. Both silkworms showed different spinning rates at different spinning stages, although the speed of the WO7q silkworm was comparatively slower in the later spinning stage. The silkworm which constructed a flossy cocoon spun primarily in a C-letter shape changed its direction frequently. On the other hand, the larva of peanut-shaped cocoons often assumed S-letter shapes and displayed a desire to change at a considerably lower frequency. Since the silkworms wO7+ and wO7q are segregated in the strain wO7 which has been maintained for more than 25 years using the same sib mating, different characteristics shown in this experiment should be considered for genetical reasons..
107. Yukiko Yamazaki, Ryo Akashi, Yutaka Banno, Takashi Endo, Hiroshi Ezura, Kaoru Fukami-Kobayashi, Kazuo Inaba, Tadashi Isa, Katsuhiko Kamei, Fumie Kasai, Masatomo Kobayashi, Nori Kurata, Makoto Kusaba, Tetsuro Matuzawa, Shohei Mitani, Taro Nakamura, Yukio Nakamura, Norio Nakatsuji, Kiyoshi Naruse, Hironori Niki, Eiji Nitasaka, Yuichi Obata, Hitoshi Okamoto, Moriya Okuma, Kazuhiro Sato, Tadao Serikawa, Toshihiko Shiroishi, Hideaki Sugawara, Hideko Urushibara, Masatoshi Yamamoto, Yoshio Yaoita, Atsushi Yoshiki, Yuji Kohara, NBRP databases
Databases of biological resources in Japan, Nucleic Acids Research, 10.1093/nar/gkp996, 38, SUPPL.1, 2009.11, The National BioResource Project (NBRP) is a Japanese project that aims to establish a system for collecting, preserving and providing bioresources for use as experimental materials for life science research. It is promoted by 27 core resource facilities, each concerned with a particular group of organisms, and by one information center. The NBRP database is a product of this project. Thirty databases and an integrated database-retrieval system (BioResource World: BRW) have been created and made available through the NBRP home page ( The 30 independent databases have individual features which directly reflect the data maintained by each resource facility. The BRW is designed for users who need to search across several resources without moving from one database to another. BRW provides access to a collection of 4.5-million records on bioresources including wild species, inbred lines, mutants, genetically engineered lines, DNA clones and so on. BRW supports summary browsing, keyword searching, and searching by DNA sequences or gene ontology. The results of searches provide links to online requests for distribution of research materials. A circulation system allows users to submit details of papers published on research conducted using NBRP resources..
108. Takashi Sakudoh, Tetsuya Iizuka, Junko Narukawa, Hideki Sezutsu, Isao Kobayashi, Seigo Kuwazaki, Yutaka Banno, Akitoshi Kitamura, Hiromu Sugiyama, Naoko Takada, Hirofumi Fujimoto, Keiko Kadono-Okuda, Kazuei Mita, Toshiki Tamura, Kimiko Yamamoto, Kozo Tsuchida, A CD36-related transmembrane protein is coordinated with an intracellular lipid-binding protein in selective carotenoid transport for cocoon coloration, Journal of Biological Chemistry, 10.1074/jbc.M109.074435, 285, 10, 7739-7751, 2010.03, The transport pathway of specific dietary carotenoids from the midgut lumen to the silk gland in the silkworm, Bombyx mori, is a model system for selective carotenoid transport because several genetic mutants with defects in parts of this pathway have been identified that manifest altered cocoon pigmentation. In the wild-type silkworm, which has both genes, Yellow blood (Y) and Yellow cocoon (C), lutein is transferred selectively from the hemolymph lipoprotein to the silk gland cells where it is accumulated into the cocoon. The Y gene encodes an intracellular carotenoid-binding protein (CBP) containing a lipid-binding domain known as the steroidogenic acute regulatory protein-related lipid transfer domain. Positional cloning and transgenic rescue experiments revealed that the C gene encodes Cameo2, a transmembrane protein gene belonging to the CD36 family genes, some of which, such as the mammalian SR-BI and the fruit fly ninaD, are reported as lipoprotein receptors or implicated in carotenoid transport for visual system. In C mutant larvae, Cameo2 expression was strongly repressed in the silk gland in a specific manner, resulting in colorless silk glands and white cocoons. The developmental profile of Cameo2 expression, CBP expression, and lutein pigmentation in the silk gland of the yellow cocoon strain were correlated. We hypothesize that selective delivery of lutein to specific tissue requires the combination of two components: 1) CBP as a carotenoid transporter in cytosol and 2) Cameo2 as a transmembrane receptor on the surface of the cells..
109. Ryo Futahashi, Yutaka Banno, Haruhiko Fujiwara, Caterpillar color patterns are determined by a two-phase melanin gene prepatterning process
New evidence from tan and laccase2, Evolution and Development, 10.1111/j.1525-142X.2010.00401.x, 12, 2, 157-167, 2010.03, The larval color patterns in Lepidoptera exhibit splendid diversity, and identifying the genes responsible for pigment distribution is essential to understanding color-pattern evolution. The swallowtail butterfly, Papilio xuthus, is a good candidate for analyzing marking-associated genes because its body markings change dramatically at the final molt. Moreover, the silkworm Bombyx mori is most suitable for identification of lab-generated color mutants because genome information and many color mutants are available. Here, we analyzed the expression pattern of 10 melanin-related genes in P. xuthus, and analyzed whether these genes were responsible for Bombyx larval color mutants. We found that seven genes correlated strongly with the stage-specific larval cuticular markings of P. xuthus, suggesting that, compared with Drosophila, more genes showed marking specificity in lepidopteran larvae. We newly found that the expression of both tan and laccase2 is strongly correlated with the larval black markings in both P. xuthus and B. mori. The results of F2 linkage analysis and mutant analysis strongly suggest that tan is the responsible gene for Bombyx larval color mutant rouge, and that tan is important in emphasizing black markings of lepidopteran larvae. Detailed comparison of temporal and spatial expression patterns showed that larval cuticular markings were regulated at two different phases. Markingspecific expression of oxidizing enzymes preceded the marking-specific expression of melanin synthesis enzymes at mRNA level, which is the reverse of the melanin synthesis step..
110. Banno Y, Shimada T, Kajiura Z, Sezutsu H. , The silkworm ― an attractive BioResource supplied by Japan , Experimental Animals, 59, 2, 139-146, 2010.04.
111. Ryusuke Niwa, Toshiki Namiki, Katsuhiko Ito, Yuko Shimada-Niwa, Makoto Kiuchi, Shinpei Kawaoka, Takumi Kayukawa, Yutaka Banno, Yoshinori Fujimoto, Shuji Shigenobu, Satoru Kobayashi, Toru Shimada, Susumu Katsuma, Tetsuro Shinoda, Non-molting glossy/shroud encodes a short-chain dehydrogenase/reductase that functions in the 'Black Box' of the ecdysteroid biosynthesis pathway, Development, 10.1242/dev.045641, 137, 12, 1991-1999, 2010.06, In insects, the precise timing of molting and metamorphosis is strictly guided by a principal steroid hormone, ecdysone. Among the multiple conversion steps for synthesizing ecdysone from dietary cholesterol, the conversion of 7-dehydrocholesterol to 5β-ketodiol, the so-called 'Black Box', is thought to be the important rate-limiting step. Although a number of genes essential for ecdysone synthesis have recently been revealed, much less is known about the genes that are crucial for functioning in the Black Box. Here we report on a novel ecdysteroidgenic gene, non-molting glossy (nm-g)/shroud (sro), which encodes a short-chain dehydrogenase/reductase. This gene was first isolated by positional cloning of the nm-g mutant of the silkworm Bombyx mori, which exhibits a low ecdysteroid titer and consequently causes a larval arrest phenotype. In the fruit fly, Drosophila melanogaster, the closest gene to nm-g is encoded by the sro locus, one of the Halloween mutant members that are characterized by embryonic ecdysone deficiency. The lethality of the sro mutant is rescued by the overexpression of either sro or nm-g genes, indicating that these two genes are orthologous. Both the nm-g and the sro genes are predominantly expressed in tissues producing ecdysone, such as the prothoracic glands and the ovaries. Furthermore, the phenotypes caused by the loss of function of these genes are restored by the application of ecdysteroids and their precursor 5β-ketodiol, but not by cholesterol or 7- dehydrocholesterol. Altogether, we conclude that the Nm-g/Sro family protein is an essential enzyme for ecdysteroidogenesis working in the Black Box..
112. Liu C, Yamamoto K, Cheng TC, Kadono-Okuda K, Narukawa J, Liu SP, Han Y, Futahashi R, Kidokoro K, Noda H, Kobayashi I, Tamura T, Ohnuma A, Banno Y, Dai FY, Xiang ZH, Goldsmith MR, Mita K, Xia QY., Repression of tyrosine hydroxylase is responsible for the sex-linked chocolate mutation of the silkworm, Bombyx mori., PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 107, 29, 12980-5, 2010.07, カイコには多数の自然突然変異があるが、本論分では卵から孵化したばかりの幼虫の体色が茶色(通常は黒色)となる変異体(遺伝子記号sch)について、色素形成のメカニズムを解析する一環として解析が行なわれた。本研究ではschのアレルであるschl(同変異体では25℃以上にすると致死する)も併用して候補遺伝子の絞り込みを行なった。候補遺伝子に対するRNAi実験、既存の他の黒色系のミュータントのレスキュー実験を含めた分析から、本変異体の形質発現はチロシンヒドロキシラーゼの活性発現の変化であることが明らかになった。.
113. MD Tofazzal Hossain, Satoshi Teshiba, Yuichi Shigeoka, Tetsuro Fujisawa, Yoji Inoko, Daisuke Sakano, Kohji Yamamoto, Yutaka Banno, Yoichi Aso, Structural properties of silkworm small heat-shock proteins:SHSP19.9 and sHSP20.8, Bioscience, Biotechnology and Biochemistry, 10.1271/bbb.100131, 74, 8, 1556-1563, 2010.09, sHSP20.8 and sHSP19.9 are silkworm small-heat shock proteins (sHSPs) comprising a number of polypeptides of molecular sizes of several tens of kilodaltons as subunits. The structural properties of sHSPs were investigated. sHSP19.9 was found to be aggregated by itself during incubation at 60 °C. Aggregation was suppressed in the presence of dithiothreitol and at high ionic strength. In contrast, sHSP20.8 was not aggregated. Aggregation of sHSP19.9 was partially suppressed by sHSP20.8 and in the presence of catalase as a target protein. Based on changes in small-angle X-ray scattering, it is possible that the molecular size of sHSP19.9 is larger than that of sHSP20.8, and that their molecular sizes increase with increasing temperature in a reversible, biphasic manner. sHSPs did not protect catalase from thermal inactivation, but protected it from precipitation by forming a soluble complex. sHSP20.8 and sHSP19.9 with dithiothreitol were stable against lyophilization, autoclaving at 120 °C, and boiling..
114. T. Fujii, T. Daimon, K. Uchino, Yutaka Banno, S. Katsuma, H. Sezutsu, T. Tamura, T. Shimada, Transgenic analysis of the BmBLOS2 gene that governs the translucency of the larval integument of the silkworm, Bombyx mori, Insect Molecular Biology, 10.1111/j.1365-2583.2010.01020.x, 19, 5, 659-667, 2010.10, The larval integument of the silkworm, Bombyx mori, is opaque because urate granules accumulate in the epidermis. Although the biosynthetic pathway of uric acid is well studied, little is known about how uric acid accumulates as urate granules in epidermal cells. In the distinct oily (od) mutant silkworm, the larval integument is translucent because of the inability to construct urate granules. Recently, we have found that the od mutant has a genomic deletion in the B. mori homologue of the human biogenesis of lysosome-related organelles complex1, subunit 2 (BLOS2) gene (BmBLOS2). Here, we performed a molecular and functional characterization of BmBLOS2. Northern blot analysis showed that BmBLOS2 was ubiquitously expressed in various tissues. We analysed the structure of a newly isolated mutant (odB) allelic to od and found a premature stop codon in the coding sequence of BmBLOS2 in this new mutation. Moreover, the translucent phenotype was rescued by the germ-line transformation of the wild-type BmBLOS2 allele into the od mutant. Our results suggest that BmBLOS2 is responsible for the od mutant phenotype and plays a crucial role in biogenesis of urate granules in the larval epidermis of the silkworm. The relationships amongst Hermansky-Pudlak syndrome (HPS) genes in mammals, granule group genes in Drosophila and translucent mutant genes in B. mori are discussed..
115. Urano K, Daimon T, Banno Y, Mita K, Terada T, Shimizu K, Katsuma S, Shimada T., Molecular defect of isovaleryl-CoA dehydrogenase in the skunk mutant of silkworm, Bombyx mori., FEBS Journal, 10.1111/j.1742-4658.2010.07832.x, 277, 21, 4452-63, 2010.11.
116. Yutaka Banno, Kazunori Yamamoto, Kazuhiro Nishikawa, Kei Tamura, Kohji Yamamoto, Yoichi Aso, Integration of the twenty-fourth and twenty-seventh linkage groups of the silkwom, Bombyx mori, Journal of Insect Biotechnology and Sericology, 79, 2, 67-70, 2010.12, The relationship between two linkage groups, 24th and 27th, of the standard linkage maps in the silkworm, Bombyx mori, was revaluated using four visible mutants, Sel, tyw (from the 24th linkage group) Xan, and l-li (from the 27th linkage group), as markers. All cross experiments indicated that the 4 marker genes were located on the same linkage group. These data indicate that the two original linkage groups should be integrated as a revised 24th linkage group..
117. Minoru Ujita, Mayumi Yamanaka, Yumiko Maeno, Koki Yoshida, Wakana Ohshio, Yoshinori Ueno, Yutaka Banno, Hiroshi Fujii, Hiroki Okumura, Expression of active and inactive recombinant soluble trehalase using baculovirus-silkworm expression system and their glycan structures, Journal of Bioscience and Bioengineering, 10.1016/j.jbiosc.2010.08.020, 111, 1, 22-25, 2011.01, Silkworm soluble trehalase was expressed as a fusion protein with an N-terminal or C-terminal hexahistidine tag in a baculovirus-silkworm expression system and assayed for enzymatic activity. Only N-terminally tagged trehalase showed a high activity. This study is the first to report in vitro functional expression of recombinant insect soluble trehalase..
118. Hiroko Tabunoki, Hiroaki Ode, Yutaka Banno, Susumu Katsuma, Toru Shimada, Kazuei Mita, Kimiko Yamamoto, Ryoichi Sato, Reiko Ishii-Nozawa, Jun ichi Satoh, BmDJ-1 is a key regulator of oxidative modification in the development of the Silkworm, Bombyx mori, PLoS One, 10.1371/journal.pone.0017683, 6, 3, 2011.03, We cloned cDNA for the Bombyx mori DJ-1 protein (BmDJ-1) from the brains of larvae. BmDJ-1 is composed of 190 amino acids and encoded by 672 nucleotides. Northern blot analysis showed that BmDJ-1 is transcribed as a 756-bp mRNA and has one isoform. Reverse transcriptase (RT)-PCR experiments revealed that the BmDJ-1 was present in the brain, fatbody, Malpighian tubule, ovary and testis but present in only low amounts in the silkgland and hemocyte of day 4 fifth instar larvae. Immunological analysis demonstrated the presence of BmDJ-1 in the brain, midgut, fatbody, Malpighian tubule, testis and ovary from the larvae to the adult. We found that BmDJ-1 has a unique expression pattern through the fifth instar larval to adult developmental stage. We assessed the anti-oxidative function of BmDJ-1 using rotenone (ROT) in day 3 fifth instar larvae. Administration of ROT to day 3 fifth instar larvae, together with exogenous (BmNPV-BmDJ-1 infection for 4 days in advance) BmDJ-1, produced significantly lower 24-h mortality in BmDJ-1 groups than in the control. 2D-PAGE revealed an isoelectric point (pI) shift to an acidic form for BmDJ-1 in BmN4 cells upon ROT stimulus. Among the factors examined for their effects on expression level of BmDJ-1 in the hemolymph, nitric oxide (NO) concentration was identified based on dramatic developmental stage-dependent changes. Administration of isosorbide dinitrate (ISDN), which is an NO donor, to BmN4 cells produced increased expression of BmDJ-1 compared to the control. These results suggest that BmDJ-1 might control oxidative stress in the cell due to NO and serves as a development modulation factor in B. mori..
119. Takashi Sakudoh, Takeharu Nakashima, Yoko Kuroki, Asao Fujiyama, Yuji Kohara, Naoko Honda, Hirofumi Fujimoto, Toru Shimada, Masao Nakagaki, Yutaka Banno, Kozo Tsuchida, Diversity in copy number and structure of a silkworm morphogenetic gene as a result of domestication, Genetics, 10.1534/genetics.110.124982, 187, 3, 965-976, 2011.03, The carotenoid-binding protein (CBP) of the domesticated silkworm, Bombyx mori, a major determinant of cocoon color, is likely to have been substantially influenced by domestication of this species. We analyzed the structure of the CBP gene in multiple strains of B. mori, in multiple individuals of the wild silkworm, B. mandarina (the putative wild ancestor of B. mori), and in a number of other lepidopterans. We found the CBP gene copy number in genomic DNA to vary widely among B. mori strains, ranging from 1 to 20. The copies of CBP are of several types, based on the presence of a retrotransposon or partial deletion of the coding sequence. In contrast to B. mori, B. mandarina was found to possess a single copy of CBP without the retrotransposon insertion, regardless of habitat. Several other lepidopterans were found to contain sequences homologous to CBP, revealing that this gene is evolutionarily conserved in the lepidopteran lineage. Thus, domestication can generate significant diversity of gene copy number and structure over a relatively short evolutionary time..
120. Kohji Yamamoto, Hirofumi Ichinose, Yoichi Aso, Yutaka Banno, Makoto Kimura, Takashi Nakashima, Molecular characterization of an insecticide-induced novel glutathione transferase in silkworm, Biochimica et Biophysica Acta - General Subjects, 10.1016/j.bbagen.2011.01.003, 1810, 4, 420-426, 2011.04, Background: The glutathione transferase (GST) superfamily is involved in the detoxification of various xenobiotics. We have identified a GST mRNA that was induced in the fat bodies of a silkworm strain exhibiting diazinon resistance and have investigated the enzyme properties of this GST. Methods: A soluble recombinant protein was overexpressed in Escherichia coli. Amino acid residues of interest were changed to alanine by site-directed mutagenesis. Results and conclusions: Phylogenetic analysis of the deduced amino acid sequence indicates that this GST belongs to an unclassified group previously reported in mosquitoes. This enzyme, named bmGSTu, has highly conserved amino acid residues, including Tyr7, Ser12 and Asn50. A recombinant bmGSTu was able to catalyze the biotranslation of glutathione with 1-chloro-2,4-dinitrobenzene, a synthetic substrate of GST. Kinetic analysis of bmGSTu mutants indicated that Tyr7, Ser12 and Asn50 are involved in enzyme function. General significance: These results support the hypothesis that bmGSTu may play a role in insecticide resistance in Bombyx mori..
121. Ken ichiro Tatematsu, Kimiko Yamamoto, Keiro Uchino, Junko Narukawa, Tetsuya Iizuka, Yutaka Banno, Susumu Katsuma, Toru Shimada, Toshiki Tamura, Hideki Sezutsu, Takaaki Daimon, Positional cloning of silkworm white egg 2 (w-2) locus shows functional conservation and diversification of ABC transporters for pigmentation in insects, Genes to Cells, 10.1111/j.1365-2443.2011.01490.x, 16, 4, 331-342, 2011.04, The white, scarlet and brown genes of Drosophila melanogaster encode three half-type ATP-binding cassette (ABC) transporters. In Drosophila, precursors of ommochromes and pteridines are transported by White/Scarlet and White/Brown heterodimers, respectively. The white egg 2 (w-2) mutant of the silkworm, Bombyx mori, has white eggs and eyes because of lack of ommochrome granules in the serosa and eyes. Here, we report that the silkworm w-2 locus encodes an ortholog of Drosophila scarlet. Our results indicate that Bombyx Scarlet forms a heterodimer with Bombyx White to transport ommochrome precursors, suggesting that formation of a White/Scarlet heterodimer and its involvement in the transport of ommochrome precursors are evolutionarily ancient and widely conserved traits in insects. Contrary to dipteran insects, white and scarlet were juxtaposed in a head-to-tail orientation in the silkworm genome, suggesting that the origin of white and scarlet was a tandem duplication of their ancestral transporter gene. In Bombyx, White is also essential for the transport of uric acid in larval epidermis. However, our results suggest that a Bombyx White/Scarlet heterodimer is not involved in this process. Our results emphasize the functional conservation and diversification of half-type ABC transporter families in insects, which may contribute to their extremely diverse color patterns..
122. Kohji Yamamoto, S. Teshiba, Y. Shigeoka, Y. Aso, Yutaka Banno, T. Fujiki, Yoshinori Katakura, Characterization of an omega-class glutathione S-transferase in the stress response of the silkmoth, Insect Molecular Biology, 10.1111/j.1365-2583.2011.01073.x, 20, 3, 379-386, 2011.06, The glutathione S-transferase (GST) superfamily is involved in detoxification of various xenobiotics. Using real-time PCR, mRNA encoding an omega-class GST of Bombyx mori (bmGSTO) was shown to be induced after exposure to various environmental stresses. A soluble form of recombinant protein (rbmGSTO) was functionally overexpressed in Escherichia coli cells and purified to homogeneity.Cys 38 and Pro 39 were found to be highly conserved in omega-class GSTs, and their roles were investigated by site-directed mutagenesis/kinetic analysis. Mutations of Cys 38 and Pro 39 residues affected the catalytic efficiency of enzymes, indicating that the presence of Cys 38 and Pro 39 residues is important for bmGSTO activity. Thus, bmGSTO could contribute to increasing the environmental stress resistance of lepidopteran insects..
123. Yukuhiro K, Iwata K, Komoto N, Tomita S, Kadono K, Kiuchi M, Itoh M, Banno Y., DNA polymorphism in Japanese population of wild mulberry silkmoth Bombyx mandarina , Int. J. Wild Silkmoth & Silk , 16, 65-68, 2011.07.
124. Takashi Kiuchi, Yutaka Banno, Susumu Katsuma, Toru Shimada, Mutations in an amino acid transporter gene are responsible for sex-linked translucent larval skin of the silkworm, Bombyx mori, Insect Biochemistry and Molecular Biology, 10.1016/j.ibmb.2011.04.011, 41, 9, 680-687, 2011.09, The sex-linked translucent (os) mutation in the silkworm, Bombyx mori, confers slightly translucent larval skin resulting from a decrease in the incorporation of uric acid into epidermal cells. By positional cloning, we narrowed a region linked to the os phenotype to approximately 157 kb located on scaffold Bm_scaf72 on the Z chromosome (chromosome 1). The region contained four gene models. Sequencing analysis revealed that one of the candidate genes had a 7-bp deletion in the coding region. We also found a 111-bp deletion or single-nucleotide substitution in the same gene using independent os mutant strains. Because all the mutations caused the generation of abnormal transcripts followed by translation of a truncated protein, we conclude that the mutation of this candidate gene is responsible for the translucent larval skin of the os mutant. Sequence analysis indicated that the gene responsible for the os mutation had homology to amino acid transporters of the solute carrier family of proteins. Our results suggest that solute carrier proteins are involved in uric acid transport in insects and other invertebrates..
125. Daimon T, Kozaki T, Niwa R, Kobayashi I, Furuta K, Namiki T, Uchino K, Banno Y, Katsuma S, Tamura T, Mita K, Sezutsu H, Nakayama M, Itoyama K, Shimada T, Shinoda T. , Precocious Metamorphosis in the Juvenile Hormone-Deficient Mutant of the Silkworm, Bombyx mori., PLoS Genet. , 10.1371/journal.pgen.1002486, 8, 3, e1002486, 2012.03.
126. Osanai-Futahashi, Mizuko, Tatematsu, Ken-ichiro, Yamamoto, Kimiko, Narukawa, Junko, Uchino, Keiro, Kayukawa, Takumi, Shinoda, Tetsuro, YUTAKA BANNO, Tamura, Toshiki, Sezutsu, Hideki, Identification of the Bombyx Red Egg Gene Reveals Involvement of a Novel Transporter Family Gene in Late Steps of the Insect Ommochrome Biosynthesis Pathway, JOURNAL OF BIOLOGICAL CHEMISTRY, 10.1074/jbc.M111.321331, 287, 21, 17706-17714, 2012.05.
127. Wei Sun, HongSong Yu, YiHong Shen, YUTAKA BANNO, ZhongHuai Xiang, Ze Zhang, Phylogeny and evolutionary history of the silkworm, SCIENCE CHINA-LIFE SCIENCES, 10.1007/s11427-012-4334-7, 55, 6, 483-496, 2012.06.
128. Fujii T, Banno Y, Abe H, Katsuma S, Shimada T, A homolog of the human Hermansky-Pudluck syndrome-5 (HPS5) gene is responsible for the oa larval translucent mutants in the silkworm, Bombyx mori, GENETICA, 10.1007/s10709-012-9694-1, 140, 10-12, 463-468, 2012.12.
129. Sakudoh T, Kuwazaki S, Iizuka T, Narukawa J, Yamamoto K, Uchino K, Sezutsu H, Banno Y, Tsuchida K, CD36 homolog divergence is responsible for the selectivity of carotenoid species migration to the silk gland of the silkworm Bombyx mori, JOURNAL OF LIPID RESEARCH, 10.1194/jlr.M032771, 54, 2, 482-495, 2013.02.
130. Wang L, Kiuchi T, Fujii T, Daimon T, Li M, Banno Y, Katsuma S, Shimada T, Reduced expression of the dysbindin-like gene in the Bombyx mori ov mutant exhibiting mottled translucency of the larval skin, GENOME, 10.1139/gen-2012-0127, 56, 2, 101-108, 2013.02.
131. Yamaguchi J, Banno Y, Mita K, Yamamoto K, Ando T, Fujiwara H, Periodic Wnt1 expression in response to ecdysteroid generates twin-spot markings on caterpillars, NATURE COMMUNICATIONS, 10.1038/ncomms2778, 4, 2013.05.
132. Lin Y, Meng Y, Wang YX, Luo J, Katsuma S, Yang CW, Banno Y, Kusakabe T, Shimada T, Xia QY, Vitellogenin Receptor Mutation Leads to the Oogenesis Mutant Phenotype "scanty vitellin" of the Silkworm, Bombyx mori, J Biol Chem., 10.1074/jbc.M113.462556, 288, 19, 13345-13355, 2013.05.
133. Banno Y, Nagasaki K, Tsukada M, Minohara Y, Banno J, Nishikawa K, Yamamoto K, Tamura K, Fujii T, Development of a method for long-term preservation of Bombyx mori silkworm strains using frozen ovaries, CRYOBIOLOGY, 10.1016/j.cryobiol.2013.03.004, 66, 3, 283-287, 2013.06.
134. Wang L, Kiuchi T, Fujii T, Daimon T, Li M, Banno Y, Kikuta S, Kikawada T, Katsuma S, Shimada T, Mutation of a novel ABC transporter gene is responsible for the failure to incorporate uric acid in the epidermis of ok mutants of the silkworm, Bombyx mori, INSECT BIOCHEMISTRY AND MOLECULAR BIOLOGY, 10.1016/j.ibmb.2013.03.011, 43, 7, 562-571, 2013.07.
135. Tabunoki H, Ono H, Ode H, Ishikawa K, Kawana N, Banno Y, Shimada T, Nakamura Y, Yamamoto K, Satoh J, Bono H, Identification of Key Uric Acid Synthesis Pathway in a Unique Mutant Silkworm Bombyx mori Model of Parkinson's Disease, PLOS ONE, 10.1371/journal.pone.0069130, 8, 7, 2013.07.
136. Fujii T, Abe H, Kawamoto M, Katsuma S, Banno Y, Shimada T, Albino (al) is a tetrahydrobiopterin (BH4)-deficient mutant of the silkworm Bombyx mori, INSECT BIOCHEMISTRY AND MOLECULAR BIOLOGY, 10.1016/j.ibmb.2013.03.009, 43, 7, 594-600, 2013.07.
137. Xu J, Kusakabe T, Yamamoto K, Suetsugu Y, Mon H, Li Z, Zhu L, Iiyama K, Banno Y, Yoshimura K, Lee JM, A novel third chromosomal locus controls susceptibility to Autographa californica multiple nucleopolyhedrovirus in the silkworm, Bombyx mori, APPLIED MICROBIOLOGY AND BIOTECHNOLOGY, 10.1007/s00253-013-5437-1, 98, 7, 3049-3058, 2014.04.
138. Fukumori H, Teshiba S, Shigeoka Y, Yamamoto K, Banno Y, Aso Y, Purification and characterization of cocoonase from the silkworm Bombyx mori., Bioscience, Biotechnology, and Biochemistry, 10.1080/09168451.2014.878215, 28, 2, 202-211, 2014.04.
139. Kaneko F, Kawashita K, Matsumura H, Katagiri C, Ogawa N, Shirai K, Banno Y, Moisture Permeability of Cocoon Shells: Application of Thermogravimetrical method to Small Biological Samples, Journal of Insect Biotechnology and Sericology, 10.11416/jibs.83.2_041, 83, 2, 41-46, 2014.06.
140. Yoda S, Yamaguchi J, Mita K, Yamamoto K, Banno Y, Ando T, Daimon T, Fujiwara H, The transcription factor Apontic-like controls diverse colouration pattern in caterpillars, NATURE COMMUNICATIONS, 10.1038/ncomms5936, 5, 4936, 2014.09.
141. Egi Y, Akitomo S, Fujii T, Banno Y, Sakamoto K, Silkworm strains that can be clearly destined towards either embryonic diapause or direct development by adjusting a single ambient parameter during the preceding generation, ENTOMOLOGICAL SCIENCE, 10.1111/ens.12073, 17, 4, 396-399, 2014.10.
142. Fujii T, Abe H, Kawamoto M, Banno Y, Shimada T, Positional cloning of the sex-linked giant egg (Ge) locus in the silkworm, Bombyx mori, INSECT MOLECULAR BIOLOGY, 10.1111/imb.12150, 24, 2, 213-221, 2015.04.
143. Masuda A, Xu J, Mitsudome T, Morokuma D, Mon H, Banno Y, Kusakabe T, Lee JM, Improvement of Endo-beta-N-acetylglucosaminidase H production using silkworm-baculovirus protein expression system, JOURNAL OF ASIA-PACIFIC ENTOMOLOGY, 10.1016/j.aspen.2015.01.006, 18, 2, 175-180, 2015.06.
144. Morokuma D, Mon H, Banno Y, Kusakabe T, Lee JM, Differential N-Glycan Modifications of Human Alpha 1-Acid Glycoprotein (α1AGP) Produced in Different Silkworm Strains using the Baculovirus Expression System, Journal of Insect Biotechnology and Sericology, 10.11416/jibs.84.2_049, 84, 2, 49-53, 2015.06.
145. Masuda A, Xu J, Mitsudome T, Nagata Y, Morokuma D, Mon H, Banno Y, Kusakabe T, Lee JM, Mass Production of an Active Peptide-N-Glycosidase F Using Silkworm-Baculovirus Expression System, MOLECULAR BIOTECHNOLOGY, 10.1007/s12033-015-9866-1, 57, 8, 735-745, 2015.08.
146. Xu J, Zhang P, Kusakabe T, Mon H, Li Z, Zhu L, Iiyama K, Banno Y, Morokuma D, Lee JM, Comparative proteomic analysis of hemolymph proteins from Autographa californica multiple nucleopolyhedrovirus (AcMNPV)-sensitive or -resistant silkworm strains during infections, COMPARATIVE BIOCHEMISTRY AND PHYSIOLOGY D-GENOMICS & PROTEOMICS, 10.1016/j.cbd.2015.07.003, 16, 36-47, 2015.12.
147. Yuasa M, Kiuchi T, Banno Y, Katsuma S, Shimada T, Identification of the silkworm quail gene reveals a crucial role of a receptor guanylyl cyclase in larval pigmentation, INSECT BIOCHEMISTRY AND MOLECULAR BIOLOGY, 10.1016/j.ibmb.2015.10.016, 68, 33-40, 2016.01.
148. Hino M, Kawanami T, Xu J, Morokuma D, Hirata K, Yamashita M, Karasaki N, Tatsuke T, Mon H, Iiyama K, Kamiya N, Banno Y, Kusakabe T, Lee JM, High-level expression and purification of biologically active human IL-2 using silkworm-baculovirus expression vector system, JOURNAL OF ASIA-PACIFIC ENTOMOLOGY, 10.1016/j.aspen.2016.03.014, 19, 2, 313-317, 2016.03.
149. Fujii T, Yamamoto K, Banno Y, Molybdenum cofactor deficiency causes translucent integument, male-biased lethality, and flaccid paralysis in the silkworm Bombyx mori., Insect Biochemistry and Molecular Biology, 10.1016/j.ibmb.2016.03.008, 73, 20-26, 2016.06.
150. Fukumori H, Fujii T, Banno Y, In vitro culture and low temperature incubation tolerance of staged embryos of the silkworm, Bombyx mori., Journal of Insect Biotechnology and Sericology, 10.11416/jibs.85.2_049, 85, 2, 49-53, 2016.06.
151. Chen J, Xu J, Hino M, Yamashita M, Hirata K, Patil AA, Tatsuke T, Mon H, Banno Y, Kusakabe T, Lee JM, Co-expression of silkworm allatostatin-C receptor BNGR-A1 with its cognate G protein subunits enhances the GPCR display on the budding baculovirus, JOURNAL OF ASIA-PACIFIC ENTOMOLOGY, 10.1016/j.aspen.2016.07.007, 19, 3, 753-760, 2016.09.
152. Fujii T, Ohnuma A, Banno Y, Abe H, Structural analysis of spontaneous Z-W translocations in the silkworm, Bombyx mori, Journal of Insect Biotechnology and Sericology,, 85, 3, 79-85, 2016.10.
153. Yutaka Banno, Tsuguru Fujii, Hisayoshi Fukumori, Kimiko Yamamoto, Jun Kobayashi, Genetic studies on two egg mutants, “small size egg” and “lethal non- diapausing egg” in the silkworm, Bombyx mori, Journal of Insect Biotechnology and Sericology, 10.11416/jibs.86.3_123, 86, 3, 123-126, 2017.01.
154. Tsuguru Fujii, Yutaka Banno, Unique phenotype of the metamorphosis-defective mutant Ishigameyoh (gap)
Establishment of a PCR-based marker for efficacious mutant maintenance in Bombyx mori, Journal of Insect Biotechnology and Sericology, 10.11416/jibs.86.3_095, 86, 3, 95-103, 2017.01, Ishigameyoh (gap) was identified as a spontaneous mutant of Bombyx mori with an apterous and sterile phenotype. Phenotypic observations revealed that the gap mutation affected the development of imaginal discs and the primordia of adult organs, including the wings, legs, compound eyes, and reproductive organs. Therefore, Ishigameyoh (gap) is a metamorphosis-defective mutant. We found that oogenesis depends on the gap gene, whereas spermatogenesis is independent. Ovary transplantation experiments suggest a cell-autonomous requirement for the gap gene in the process of oogenesis. The gap mutation has been maintained by crossing +gap/ gap moths as the u90 strain. However, the u90 strain is difficult to maintain because +gap/+gap and +gap/gap moths cannot be distinguished phenotypically. For efficacious maintenance, we established a molecular marker that is closely linked to the gap locus. First, we localized the gap locus to chromosome 5 using phenotypic markers. Second, the gap locus was narrowed down to within a 1-Mb region using PCR-based markers. Within this 1-Mb region, we finally established a PCR-based marker that amplified different-sized products from +gap/+gap and gap/ gap offspring of the u90 strain. This marker allows the distinction of +gap/+gap and +gap/gap moths with an error rate of <1% and therefore reduces the labor needed to maintain the u90 strain. Moreover, this marker helps to identify +gap and gap individuals before pupation, which is necessary for larvae-based genetic and physiological analyses of the gap gene..
155. Yamashita M, Xu J, Morokuma D, Hirata K, Hino M, Mon H, Takahashi M, Hamdan SM, Sakashita K, Iiyama K, Banno Y, Kusakabe T, Lee JM. , Characterization of Recombinant Thermococcus kodakaraensis (KOD) DNA Polymerases Produced Using Silkworm-Baculovirus Expression Vector System, MOLECULAR BIOTECHNOLOGY, 10.1007/s12033-017-0008-9, 59, 6, 221-233, 2017.06.
156. Fukumori H, Lee J, Fujii T, Kajiura Z, Banno Y., Long-term preservation of eri and ailanthus silkworms using frozen gonads, CRYOBIOLOGY, 10.1016/j.cryobiol.2017.05.003, 77, 71-74, 2017.08.
157. KonDo Y, Yoda S, Mizoguchi T, Ando T, Yamaguchi J, Yamamoto K, Banno Y, Fujiwara H., Toll ligand Spatzle3 controls melanization in the stripe pattern formation in caterpillars, PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 10.1073/pnas.1707896114, 114, 31, 8336-8341, 2017.08.
158. Zhang H, Kiuchi T, Wang L, Kawamoto M, Suzuki Y, Sugano S, Banno Y, Katsuma S, Shimada T., Bm-muted, orthologous to mouse muted and encoding a subunit of the BLOC-1 complex, is responsible for the otm translucent mutation of the silkworm Bombyx mori, GENE, 10.1016/j.gene.2017.07.071, 629, 92-100, 2017.09.
159. Kazunori Matsuo, Yoshimi Hirose, Takeshi Yokoyama, Yumiko Nakajima, Yu Feng Hsu, Yutaka Banno, Discovery of a New Species of Telenomus (Hymenoptera
Scelionidae) Parasitic on Eggs of Bombyx mandarina and Bombyx mori (Lepidoptera: Bombycidae) in Japan and Taiwan, Journal of Insect Science, 10.1093/jisesa/iey072, 18, 4, 2018.07, We reared a Telenomus species from eggs of Bombyx mandarina (Moore) (Lepidoptera: Bombycidae) and Bombyx mori (Linnaeus) (Lepidoptera: Bombycidae) in Japan, and from eggs of B. mandarina in Taiwan. Morphological examination revealed that this Telenomus species is new to science. In this article, we describe it as Telenomus moricolus Matsuo et Hirose, sp. nov. Because B. mandarina is considered to be an ancestor of B. mori, a domestic insect, it is reasonable to assume that B. mandarina is an original host of T. moricolus. This is the second discovery of an egg parasitoid attacking wild and domesticated silkworms, following the first discovery of T. theophilae, a Chinese species. The significance of the discovery of T. moricolus is discussed in relation to examining the effects of host-insect domestication on egg parasitism..
160. Tsuguru Fujii, Kazunori Yamamoto, Yutaka Banno, Translucent larval integument and flaccid paralysis caused by genome editing in a gene governing molybdenum cofactor biosynthesis in Bombyx mori, Insect Biochemistry and Molecular Biology, 10.1016/j.ibmb.2018.04.008, 99, 11-16, 2018.08, Translucency of the larval integument in Bombyx mori is caused by a lack of uric acid in the epidermis. Hime'nichi translucent (ohi) is a unique mutation causing intermediate translucency of the larval integument and male-specific flaccid paralysis. To determine the gene associated with the ohi mutation, the ohi locus was mapped to a 400-kb region containing 29 predicted genes. Among the genes in this region, we focused on Bombyx homolog of mammalian Gephyrin (BmGphn), which regulates molybdenum cofactor (MoCo) biosynthesis, because MoCo is indispensable for the activity of xanthine dehydrogenase (XDH), a key enzyme in uric acid biosynthesis. The translucent integument of ohi larvae turned opaque after injection of bovine xanthine oxidase, which is a mammalian equivalent to XDH, indicating that XDH activity is defective in ohi larvae. RT-PCR and sequencing analysis showed that (i) in ohi larvae, expression of the BmGphn gene was repressed in the fat body where uric acid is synthesized, and (ii) there was no amino acid substitution in the ohi mutant allele. Finally, we obtained BmGphn knockout alleles (hereafter denoted as BmGphnΔ) by using CRISPR/Cas9. The resulting ohi/BmGphnΔ larvae had translucent integuments, demonstrating that BmGphn is the gene responsible for the ohi phenotype. Our results show that repressed expression of BmGphn is a causative factor for the defective MoCo biosynthesis and XDH activity observed in ohi larvae. Interestingly, all male BmGphnΔ homozygotes died before pupation and showed a flaccid paralysis phenotype. The genetic and physiological mechanisms underlying this flaccid paralysis phenotype are also discussed..
161. Haokun Zhang, Takashi Kiuchi, Chikara Hirayama, Yutaka Banno, Susumu Katsuma, Toru Shimada, A reexamination on the deficiency of riboflavin accumulation in Malpighian tubules in larval translucent mutants of the silkworm, Bombyx mori, Genetica, 10.1007/s10709-018-0034-y, 146, 4-5, 425-431, 2018.10, A variety of insects accumulate high contents of riboflavin (vitamin B2) in their Malpighian tubules (MTs). Although this process is known to be genetically controlled, the mechanism is not known. In the 1940s and the 1950s, several studies showed that riboflavin contents were low in the MTs of some Bombyx mori (silkworm) mutants with translucent larval skin mutations (e.g., w-3, od, oa, and otm) and that genes responsible for these translucent mutations also affected riboflavin accumulation in the MTs. Since the 2000s, it has been shown that the w-3 gene encodes an ABC transporter, whereas genes responsible for od, oa, and otm mutations encode for the biogenesis of lysosome-related organelles. These findings suggest that some genes of ABC transporters and biogenesis of lysosome-related organelles may control the accumulation of riboflavin in MTs. Therefore, we reexamined the effects that translucent mutations have on the accumulation of riboflavin in MTs by using the translucent and wild-type segregants in mutant strains to measure the specific effect that each gene has on riboflavin accumulation (independent of genomic background). We used nine translucent mutations (w-3oe, oa, od, otm, Obs, oy, or, oh, and obt) even though the genes responsible for some of these mutations (Obs, oy, or, oh, and obt) have not yet been isolated. Through observation of larval MTs and measurements of riboflavin content using high-performance liquid chromatography, we found that the oa, od, otm, and or mutations were responsible for low contents of riboflavin in MTs, whereas the Obs and oy mutations did not affect riboflavin accumulation. This indicates that the molecular mechanism for riboflavin accumulation is similar but somewhat different than the mechanism responsible for uric acid accumulation in epidermal cells. We found that the genes responsible for oa, od, and otm mutations were consistent with those already established for uric acid accumulation in larval epidermis. This suggests that these three genes control riboflavin accumulation in MTs through a mechanism similar to that of uric acid accumulation, although we do not yet know why the or mutation also controls riboflavin accumulation..
162. Lingyan Wang, Zhaoming Dong, Juan Wang, Yaru Yin, Huawei Liu, Wenbo Hu, Zhangchuan Peng, Chun Liu, Muwang Li, Yutaka Banno, Toru Shimada, Qingyou Xia, Ping Zhao, Proteomic Analysis of Larval Integument in a Dominant Obese Translucent (Obs) Silkworm Mutant, Journal of Insect Science, 10.1093/jisesa/iey098, 18, 6, 2018.11, The dominant obese translucent (Obs) mutant of the silkworm (Bombyx mori) results in a short and stout larval body, translucent phenotype, and abnormal pigmentation in the integument. The Obs mutant also displays deficiency in ecdysis and metamorphosis. In the present study, to gain an understanding of multiple Obs phenotypes, we investigated the phenotypes of Obs and performed a comparative analysis of the larval integument proteomes of Obs and normal silkworms. The phenotypic analysis revealed that the Obs larvae were indeed short and fat, and that chitin and uric acid content were lower but melanin content was higher in the Obs mutant. Proteomic analysis revealed that 244 proteins were significantly differentially expressed between Obs and normal silkworms, some of which were involved in uric acid metabolism and melanin pigmentation. Twenty-six proteins were annotated as cuticular proteins, including RR motif-rich cuticular proteins (CPR), glycine-rich cuticular protein (CPG), hypothetical cuticular protein (CPH), cuticular protein analogous to peritrophins (CPAPs), and the chitin_bind_3 motif proteins, and accounted for over 84% of the abundance of the total significantly differentially expressed proteins. Moreover, 22 of the 26 cuticular proteins were downregulated in the Obs mutant. Comparative proteomic analysis suggested that the multiple phenotypes of the Obs mutant might be related to changes in the expression of proteins that participate in cuticular formation, uric acid metabolism, and melanin pigmentation. These results could lay a basis for further identification of the gene responsible for the Obs mutant. The data have been deposited to ProteomeXchange with identifier PXD010998..
163. Tsuguru Fujii, Yutaka Banno, Identification of a novel function of the silkworm integument in nitrogen metabolism
Uric acid is synthesized within the epidermal cells in B. mori, Insect Biochemistry and Molecular Biology, 10.1016/j.ibmb.2018.12.014, 105, 43-50, 2019.02, During nitrogen metabolism, animals convert toxic ammonia to less toxic forms. Uric acid (UA) is an end product of this process in terrestrial insects. In lepidopteran larvae, a large amount of UA is stored in the integument via a phenomenon known as storage excretion. Physiologically, integumental UA plays crucial roles as a barrier against sunlight and as a white pigment for larval pigmentation patterns. Conventionally, UA is thought to be synthesized in the fat body, the insect equivalent of the liver of vertebrates, and to be transported to the epidermis via the hemolymph. Here, we reconsidered the conventional theory by a mosaic analysis targeting genes governing UA synthesis, using CRISPR/Cas9 mutagenesis and a traditional genetic method in Bombyx mori. Notably, we observed mosaic larvae in which the integument comprised both UA-containing white and UA-lacking translucent areas, indicating that UA synthesis in the epidermis is indispensable to the accumulation of a large amount of highly insoluble UA in the epidermis. Our results thus provide a genetic basis for storage excretion wherein lepidopteran insects use nitrogenous waste to adapt to their environment..