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Fine-mapping reveals novel alternative splicing of BmAbd-A in Silkworm E Mutant

Fine-mapping reveals novel alternative splicing of BmAbd-A i...

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第十届家（柞）蚕遗传育种及良种繁育学术研讨会

作者： Dandan Li Xiaoling Tong Peng Chen Pingfeng Liang Mingyue Fu Cheng Lu fangyin dai State Key Laboratory of Silkworm Genome Biology Key Laboratory for Sericulture Functional Genomics and Biotechnology of Agricultural MinistrySouthwest University

The versatility of appendages in insects plays an important role in evolution and *** of these appendages is regulated by a complex set of genes including the Hox gene. To study the mechanism of Hox gene action on insect body plan, the E group of silkworm with more than 30 mutants is an excellent model. In silkworm, Abd-A that plays an important role in development of prolegs, and is expressed in most of the abdomen. In this study, we identified multiple novel alternative splice forms of Abd-A in E mutant silkworms. To investigate the contributions of Abd-A splice variants to prolegs development,we analyzed the expression levels of Abd-A mRNA and protein in E mutant Bombyx *** addition, we also analyzed the Distal-less（Dll） which is the downstream target gene of Abd-A, during embryogenesis. Further, we also performed a detailed analysis on the evolutionary pattern of the Abd-A alternative splice forms in diverse species. Together, our results suggest that（1） multiple spliceosomes may induce the over-expression of Abd-A and promote the expression of Dll for abdominal appendage development（2） successful evolution of Abd-A gene duplication and various splicing forms was adapted by organisms to perform complex and multiplex functions in vivo.

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Cyclin B3 plays pleiotropic roles in female reproductive organogenesis and early embryogenesis in the silkworm,Bombyx mori

Cyclin B3 plays pleiotropic roles in female reproductive org...

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中国蚕学会2023年学术年会

作者： Hai Hu Duan Tan Tianfu Luo Xiaoling Tong Minjin Han Jianghong Shen fangyin dai

BACKGROUND The reproductive system plays a crucial role in insect survival,reproduction and species *** the molecular mechanisms underlying reproductive organogenesis contributes to improving the efficiency of sterile insect technique marked by an eco-friendly pest management *** is one of the largest orders of insects,most of which are major pests in agriculture and *** study aimed to screen the genes responsible for reproductive organogenesis and unravel the mechanism underlying female reproductive organ *** Morphological investigation of female reproductive organs showed a defective connection between oviductus geminus and oviductus communis on the second day of pupa（P2） in Speckled mutant *** identified a total of 18 049 transcripts that were expressed in the P2 female internal reproductive organs without ovary in Spc/+ compared to +Spc/+*** expression analysis identified 312 upregulated genes and 221 down-regulated genes in Spc/+.KEGG analysis identified 44 significantly enriched *** results of qRT-PCR performed on 33 genes significantly matched the outcomes of the *** of Cyclin B3 resulted in a defective connection of the oviductus communis with the ovariole,dysfunction of oogenesis,and a petite ***,homozygous recessive lethality of Cyclin B3/Cyclin B3 occurred during early *** Our results suggested that Cyclin B3 is a pleiotropic functional gene that regulates early embryogenesis,oogenesis,development,and female reproductive *** results showed that Cyclin B3 has significant effects on lepidopteran mortality,growth,and reproductive physiology,which might be considered a novel and potentially eco-friendly target for lepidopteran pest management.

关键词： transcriptome CRISPR/Cas9 Cyclin B3 female internal reproductive organogenesis embryogenesis Bombyx mori targets of pest control

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A novel laminin β gene BmLanB1-w regulates wing-specific cell adhesion in silkworm, Bombyx mori

A novel laminin β gene BmLanB1-w regulates wing-specific ce...

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第十二届家（柞）蚕遗传育种暨良种繁育学术研讨会

作者： Xiaoling Tong Songzhen He Jun Chen Hai Hu Zhonghuai Xiang Cheng Lu fangyin dai State Key Laboratory of Silkworm Genome Biology Key Laboratory for Sericulture Functional Genomics and Biotechnology of Agricultural Ministry Southwest University

Laminins are important basement membrane（BM） components with crucial roles in *** numbers of laminin isoforms in various organisms are determined by the composition of the different α,β,and γ chains,and their coding genes,which are variable across *** insects,only two α,one β,and one γ chains have been identified thus ***,we isolated a novel laminin β gene,BmLanB1-w,by positional cloning of the mutant（crayfish,cf） with blistered wings in *** structure analysis showed that a 2 bp deletion of the BmLanB1-w gene in the cf mutant caused a frame-shift in the open reading frame（ORF） and generated a premature stop *** of the BmLanB1-w gene produced individuals exhibiting blistered wings,indicating that this laminin gene was required for cell adhesion during wing *** also identified laminin homologs in different species and showed that two copies of β laminin likely originated in Lepidoptera during ***,phylogenetic and gene expression analyses of silkworm laminin genes revealed that the BmLanB1-w gene is newly evolved,and is required for wing specific cell *** is the first report showing the tissue specific distribution and functional differentiation of β laminin in insects.

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Sample Preparation to Observe the Straight and Flat Posture of Silkworm Embryo under Scanning Electron Microscopy via Glycerol Substitution Method

Sample Preparation to Observe the Straight and Flat Posture ...

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第十二届家（柞）蚕遗传育种暨良种繁育学术研讨会

作者： Wen bin Liu Chenzhao Li Qi Zhang Zhix in Lei Yan qi Hou fangyin dai Cheng Lu Department of Pharmaceutical and Biological Engineering School of Chemical Engineering Sichuan University State Key Laboratory of Silkworm Genome Biology Southwest University

In the preparation process for scanning electron microscopy（SEM）, flexed silkworm embryos typically assume several curled shapes with irregular postures that obscure morphological details during SEM observation. We describe a preparation technique based on glycerol substitution for better SEM visualization of straight and flat silkworm embryos. Glycerol has high viscosity, low vapor pressure, and sufficient electrical conductivity. Silkworm embryos were infiltrated with glycerol and arranged in astraight posture or flattened using a cover slip. Samples were directly observed by SEM without additional dehydration, drying, or coating procdures. The complete ventral side could be easily viewed in one image. Recoating alleviated the charging phenomenon. This represent s a simple method for preparation of straight and flat samples from curled biological specimens for SEM observation.

关键词： SEM preparation silkworm embryo glycerol substitution

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Comparative Analysis of the Integument Transcriptomes between Stick Mutant and Wild-Type Silkworms

Comparative Analysis of the Integument Transcriptomes betwee...

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中国蚕学会第十届青年学术研讨会

作者： Duan Tan Hai Hu Xiaoling Tong Minjin Han Songyuan Wu Xin Ding fangyin dai Cheng Lu State Key Laboratory of Silkworm Genome Biology Key Laboratory of Sericultural Biology and Genetic BreedingMinistry of AgricultureCollege of BiotechnologySouthwest University

In insects,the integument provides mechanical support for the whole body and protects them from infections,physical and chemical injuries,and *** in integument properties is often related to body shape,behavior,and survival *** stick（sk） silkworm is a spontaneous mutant with a stick-like larval body that is firm to the touch and,thus,less *** of the mechanical properties of the cuticles at day 3 of the fifth instar（L5D3） of sk larvae revealed higher storage modulus and lower loss *** sequencing identified a total of 19,969 transcripts that were expressed between wild-type Dazao and the sk mutant at L5D2,of which 11,596 transcripts were novel and detected in the *** expression analyses identified 710 upregulated genes and 1009 downregulated genes in the sk *** Ontology（GO） enrichment analysis indicated that four chitin-binding peritrophin A domain genes and a chitinase gene were upregulated,whereas another four chitin-binding peritrophin A domain genes,a trehalase,and nine antimicrobial peptides were *** Encyclopedia of Genes and Genomes（KEGG） analysis indicated that two functional pathways,namely,fructose and mannose metabolism and tyrosine metabolism,were significantly enriched with differentially-expressed *** study provides a foundation for understanding the molecular mechanisms underlying the development of the stiff exoskeleton in the sk mutant.

关键词： Bombyx mori stick mutant transcriptome integument stiff exoskeleton

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Fine Mapping of a Degenerated Abdominal Legs Mutant（Edl） in Silkworm,Bombyx mori

Fine Mapping of a Degenerated Abdominal Legs Mutant（Edl） i...

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中国蚕学会第九届青年学术研讨会

作者： Honglei Wang Xiaoling Tong Meijing Liu Hai Hu Zhiquan Li Zhonghuai Xiang fangyin dai Cheng Lu State Key Laboratory of Silkworm Genome Biology Key Laboratory for Sericulture Functional Genomics and Biotechnology of Agricultural MinistrySouthwest University

In insects,abdominal appendages,also called prolegs,vary due to adaptive *** on prolegs within species provide insights to better understand the mechanisms underlying appendage development and *** silkworm Bombyx mori,extra-crescents and degenerated abdominal legs（Edl） mutant,belonging to the E pseudoallele group,is a spontaneous mutation that adds crescents and degenerates prolegs on the third abdominal segment（A3）.This mutation may be a homeotic transformation of A3 to *** this study,the Edl locus was mapped within approximately a 211 Kb region that is 10 Kb upstream of Bmabdominal-A（Bmabd-A）.RT-quantitative PCR（RT-qPCR） and Western blot analysis of Bmabd-A expression showed a slight but significant decrease,while the expression of BmUltrabithorax（BmUbx） was up-regulated in the Edl mutant compared to wildtype（Dazao）.Moreover,we also found that BmDistal-less（BmDll）,which regulated the development of distal proleg structures,was missing at the tips of the A3 prolegs in the Edl mutant compared to BmDll expression in normally developed prolegs in both the wildtype and ***,we identified approximately a 211 Kb region in the Edl locus that regulates BmUbx and Bmabd-A expression and found that changes in BmUbx and Bmabd-A expression may lead to the loss of distal proleg structures in ***.

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The regulation of Hox gene expression during silkworm development

The regulation of Hox gene expression during silkworm develo...

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中国蚕学会第八届青年学术研讨会

作者： Xiaoling Tong fangyin dai Peng Chen Mingyue Fu Yaqun Xin Lei Chen Zhonghuai Xiang Cheng Lu State Key Laboratory of Silkworm Genome Biology Key Laboratory for Sericulture Functional Genomics and Biotechnology of Agricultural Ministry Southwest University

Hox genes encoding homeodomain-containing transcription factors control the embryogenesis and the specification of various tissue structures during animal development. The specific regional functions of individual Hox genes largely reflect their restricted expression patterns, the disruption of which can lead to developmental defects and disease. In silkworm, there are more than 30 mutants（E group） exhibiting multiple phenotypes, such as extra color markings, supernumerary or defective legs, abnormal silkgland, defective wings and embryonic lethality. Here, we identified the mutation region responsible for the E mutants by positional clone. The results showed that they are related to the defect of Hox gene cluster, mainly caused by mutation of the Antp,Ubx, abd-A and Abd-B. The mutations are located both in the gene coding region, which destroy the encoding protein, and in the intergenic region, which affect Hox gene expression patterns. The long non-coding RNAs（lnc RNAs） and mi RNAs were also been identified in the intergenic region between hox genes, which supposed to implicate in the control of Hox gene expression. By analyzed the role of Hox genes in the tissues differentiation in silkworm, some new findings got in the silkworm were opposed to previous reports in Drosophila and other insects. For instance, the Antp gene was found to control the wing development in silkworm,while the Antp gene is not affect the wings identity in Drosophila and Tribolium castaneum. In silkworm, the Abd-A gene promotes the proleg development. Ectopic expressed Abd-A gene in the A2 segment actives the ectopic dll expression and thereby induces extral prolegs. However, the Abd-A gene inhibits the leg development by repressing dll expression in Drosophila. The research on Hox genes and their roles in establishing the body structures in silkworm will improve our understanding the regulatory mechanisms of Hox gene during vertebrate development.

关键词： silkworm Hox gene development E mutants

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Fine Mapping of a Degenerated Abdominal Legs Mutant（Edl） in Silkworm,Bombyx mori

Fine Mapping of a Degenerated Abdominal Legs Mutant（Edl） i...

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中国蚕学会第十届青年学术研讨会

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Fam114A2:From Silkworm Body Shape to Human Tumor Repressor

Fam114A2:From Silkworm Body Shape to Human Tumor Repressor

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中国蚕学会第十届青年学术研讨会

作者： Xiaoling Tong Niannian Li Fuze Sun Gang Meng Jie Zeng Hai Hu Cheng Lu fangyin dai State Key Laboratory of Silkworm Genome Biology Key Laboratory for Sericulture Biology and Genetic BreedingMinistry of Agriculture and Rural AffairsCollege of BiotechnologySouthwest University

As a fascinating economic insect,silkworm also has been successfully used as a model organism in biological research in the past decade due to several features,including completed silkworm genome,clear genetic background and rich genetic ***,the genome sequencing of silkworm reveals that approximately 58% of silkworm genes have orthologs in ***,silkworm is a potential animal model for understanding of human genetic *** this article,using positional cloning,we identified the Bmfam114a2 gene as one of the causal gene responsible for the Squab（Sq） mutant,exhibiting short and fat body in *** silkworm,fam114a2 RNAi caused the Squab-like body shape in the embryonic *** cell proliferation was increased by down-regulating the fam114a2 gene expression in the silkworm embryonic cell line BmE,and the Sq mutant embryos also presented over-proliferation compared to wild type using EDU *** the tumors exhibit a condition in which cell proliferation is uncontrolled and malignant proliferation occurs,subsequently,we test the homologue gene of fam114a2 in *** F114a2 expressed at a lower level in the tissues of kidney cancer,gastric cancer and liver cancer compared that in the adjacent tissues of corresponding *** kidney cancer as the main research object,F114a2 was found to inhibit tumor formation and development in vitro and in *** found that f114a2 inhibits cell proliferation by inducing G1/S phase cell cycle *** passage of cells from the G1 phase to the S phase requires two cell cycle checkpoints.F114a2 affects the formation of the first cycle checkpoint cyclinD1/cdk4 by interaction with p19,akt,and β-catenin,and then interacts with p21,p27,and p53 to affect the second cell cycle checkpoint CyclinE/***,we used AAV virus as a toolkit to treat mouse kidney cancer by overexpressed *** results showed that F114a2 is a potential target for gene therapy of kidney tumor diseases

关键词： Silkworm Sq mutant Cell cycle Cell proliferation Tumor

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Cuticular protein defective Bamboo mutant of Bombyx mori is sensitive to environmental stresses

Cuticular protein defective Bamboo mutant of Bombyx mori is ...

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中国蚕学会第十四届暨国家蚕桑产业技术体系家（柞）蚕遗传育种学术研讨会

作者： Gao Xiong Xiaoling Tong Zhengwen Yan Hai Hu Xiaohui Duan Chunlin Li Minjin Han Cheng Lu fangyin dai State Key Laboratory of Silkworm Genome Biology Southwest University Key Laboratory of Sericultural Biology and Genetic Breeding Ministry of Agriculture College of Biotechnology Southwest University

Insect cuticle acts as a primary protective barrier against environment stresses that may directly impact the insect body. Here, we report the mechanical defense function of a structural cuticular protein, BmorCPH24, to environmental stresses using a silkworm Bamboo mutant(Bo) with this gene mutation. Ultraviolet(UV) irradiation and topical application of an acetone insecticide were used as environmental stresses to determine the differences in susceptibility between Bo and wild-type larvae. UV irradiation resulted in a sunburn phenotype in the Bo strains earlier than the wild-type indicating the sensitivity of Bo. Higher malondialdehyde(MDA) content and a lower survival ratio were also observed in the Bo strains. Treatment with deltamethrin revealed that Bo larvae were more sensitive to insecticides than the wild-type. Furthermore, cuticle analysis by microsection revealed thinner cuticle and a significant decrease in the endocuticle layer(～64.0%) in Bo. These results suggest that BmorCPH24 mutation can lead to deficiency in resources required to construct the cuticle in Bo resulting in thin cuticle and reduced resistance to UV and insecticides. These results provide us new insight into the role of structural cuticular proteins in insect cuticle against environment stresses.

关键词： Bombyx mori UV Insecticide resistance Cuticular protein Cuticle thickness

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