Embryonic diapause, a state of developmental arrest in silkworm (Bombyx mori) eggs, poses a challenge for year-round sericulture. While physical stimuli like corona discharge can effectively terminate diapause, the underlying molecular mechanisms, particularly the initial events, remain poorly understood. This study employed an integrated transcriptomic and proteomic approach to analyze silkworm eggs within 48 h after corona treatment. Our time-series analysis revealed that the Hippo and Wnt signaling pathways were specifically activated as early as 1 h post-treatment, preceding the previously reported FoxO pathway response. We identified two temporally distinct gene clusters within the Hippo pathway, including immediate-early genes (e.g., Dachs_17/25/29, Ft_10) and late-phase effector genes, coordinating the exit from cell cycle arrest. Concurrently, the Wnt pathway was rapidly initiated, marked by the sustained upregulation of key regulators Notum and Pontin52, suggesting its role in unlocking the cell cycle. We propose a synergistic model wherein corona discharge triggers the concurrent, early activation of Hippo and Wnt signaling, which collectively reprogram the cell cycle and reinstate the developmental trajectory by promoting proliferation and suppressing apoptosis. These findings provide crucial insights into the initial molecular events of diapause termination, identifying Hippo and Wnt pathways as master regulators in transducing the physical corona stimulus into a developmental signal.

Embryonic diapause is a reversible arrest of embryonic development that enables insects to survive adverse environmental conditions. In the silkworm (Bombyx mori), diapause is established during early egg development and can be efficiently terminated by corona discharge; however, the transcriptional responses associated with this physical stimulus remain insufficiently characterized. Here, we present a strand-specific RNA-seq dataset generated from diapause-destined silkworm eggs subjected to early or late corona discharge treatment. The dataset comprises 36 libraries from three experimental groups across eight developmental time points, with two biological replicates per condition. In total, 283.9 Gb of high-quality clean data were obtained, with average Q20 and Q30 values of 97.60% and 94.20%, respectively, and an average genome-mapping rate of 82.62%. The dataset includes raw reads, gene-expression matrices, lists of differentially expressed genes, novel transcript annotations, alternative splicing events, and SNP/InDel calls. This resource provides a time-resolved transcriptomic framework for investigating physical-stress-induced diapause termination and supports comparative studies of diapause regulation in silkworms and other insects.

The cocoon coat of Bombyx mori contains abundant bioactive proteins that contribute to its protective functions, yet varietal differences remain largely unexplored. In this study, water-soluble proteins were extracted from N4 (yellow), D9L (green), and 305 (white) cocoon coats using PBS (pH 7.0), followed by analyses of antioxidant capacity, antibacterial activity, and proteomic composition. All extracts exhibited strong antioxidant activity with DPPH scavenging rates of 71-81%. Antibacterial assays revealed distinct inhibition patterns: 305 proteins were most effective against Escherichia coli and Beauveria bassiana, D9L proteins strongly inhibited Trichophyton rubrum, while N4 showed intermediate effects. DIA-based proteomics identified 462 proteins, approximately 60% of which had catalytic or binding functions, enriched in protease and protease-inhibitor categories. Major components included Ser3 and Ser1-like sericins, low-abundance Ser2, and fibroin subunits (Fib-H, Fib-L). High-abundance proteins mainly comprised serine and carboxypeptidase inhibitors and several TIL domain-containing zonadhesin-like proteins. Differential protein analysis linked varietal antimicrobial differences to these bioactive components. Structural prediction of the zonadhesin-like protein using AlphaFold 3 revealed a unique convex-loop conformation formed by interactions among tandem TIL domains, suggesting potential roles in pathogen recognition or defense. This study establishes a comprehensive proteomic atlas of cocoon coats from diverse silkworm varieties, identifies BmSPI39 and seroin as key drivers of strain-specific defenses, and elucidates an adaptive defense mechanism mediated by soluble silk proteins, thereby providing specific molecular targets for disease-resistant silkworm breeding.

Background:To adapt to environmental changes, diapausing silkworm eggs remain dormant during the early stages of embryonic development. Various methods have been used to terminate silkworm egg diapause and promote egg hatching. Methods:To elucidate the molecular mechanisms by which corona and other treatments terminate silkworm egg diapause, we collected eggs at 1, 6, and 20 h after treatments and sequenced their transcriptomes. Results:The results showed that both corona and hydrochloric acid (HCl) treatments effectively terminated diapause and promoted egg hatching, with corona treatment inducing faster hatching than HCl treatment. Differentially expressed gene (DEG) analysis revealed the presence of fewer DEGs at 1 h, with a marked increase observed at 6 and 20 h post treatment. Functional enrichment analysis showed that the FoxO signaling pathway was activated at 6 h, with more substantial gene expression changes observed after corona treatment. In addition, HCl treatment appeared to activate the heat shock protein and hormone-regulated pathways. Our study results provide a basis for further analysis of the molecular mechanisms underlying diapause termination in silkworm eggs.

The silkworm (Bombyx mori) is rich in germplasm resources, including thermotolerant strains that live in tropical/subtropical humid climates. In this study, two thermotolerant strains and one sensitive strain were used as materials, with the former exhibiting higher critical thermal maximum (CTmax) values. Under different temperature and humidity stresses, physiological and transcriptomic responses of the fifth instar larvae were compared. It was confirmed that high humidity exacerbates harmful effects only under high temperature conditions. Based on transcriptome and co-expression network analysis, 88 evolved thermoplastic genes (Evo_TPGs) and 1338 evolved non-plastic genes (Evo_non-PGs) were identified, which exhibited specific responses or expressions in the two thermotolerant strains. Eighteen of the Evo_TPGs encode cuticular proteins, 17 of which were specifically downregulated in the two thermotolerant strains after short-term exposure to 35 °C. This may promote cuticular transpiration to dissipate internal heat, thus compensating for the suppression of tracheal ventilation in hot and humid climates. For the Evo_non-PGs, most of the metabolic genes showed lower expression at background levels in the thermotolerant strains, while oxidative stress genes showed the opposite trend, suggesting that silkworms can enhance heat tolerance by suppressing metabolic rates and allocating more resources to overcome heat-induced oxidative damage. Furthermore, the heat resistance-related genes showed higher single nucleotide polymorphisms (SNPs) between resistant and sensitive strains compared to randomly selected genes, suggesting that they may have been subjected to natural selection. Through long-term adaptive evolution, thermotolerant silkworms may reduce their internal temperature by dynamically regulating cuticle respiration in response to high temperature and humidity, while allocating more energy to cope with and repair heat-induced damage. Overall, these findings provide insights into the evolution of heat-resistant adaptations to climate change in insects.

This study performed mitochondrial genome sequencing on Minettia omei, which belongs to Lauxaniinae, a subfamily of Lauxaniidae. The complete mitochondrial genome spans 16,201 bp and comprises 37 genes, including 13 protein-coding genes, 22 tRNA genes, 2 rRNA genes, and a control region. Base composition: A 38.81%, T 38.81%, C 12.93%, G 9.44% , A + T bias 77.62%. COI uses start codon CAA; COI, COII, NAD2, NAD1, NAD4 have incomplete T + tRNA stop codons. Phylogenetic analysis elucidates the species' evolutionary relationships, establishing a critical genetic foundation for future studies on Lauxaniidae.

Probiotics have attracted considerable attention in animal husbandry due to their positive effect on animal growth and health. This study aimed to screen candidate probiotic strain promoting the growth and health of silkworm and reveal the potential mechanisms. A novel probiotic Pediococcus pentosaceus strain (ZZ61) substantially promoted body weight gain, feed efficiency, and silk yield. These effects were likely mediated by changes in the intestinal digestive enzyme activity and nutrient provisioning (e.g., B vitamins) of the host, improving nutrient digestion and assimilation. Additionally, P. pentosaceus produced antimicrobial compounds and increased the antioxidant capacity to protect the host against pathogenic infection. Furthermore, P. pentosaceus affected the gut microbiome and altered the levels of gut metabolites (e.g., glycine and glycerophospholipids), which in turn promotes host nutrition and health. This study contributes to an improved understanding of the interactions between probiotic and host and promotes probiotic utilization in sericulture.

The safety of transgenic technology is a major obstacle in the popularization and use of transgenic silkworms and their products. In sericulture, only the first filial generation (F-1) hybrid eggs produced by cross-breeding Japanese and Chinese original strains are usually used for the large-scale breeding of silkworms, but this may result in uncontrolled transgene dispersal during the popularization and application of the F-1 hybrid transgenic eggs. To address this issue, we developed a safe and efficient strategy using the GAL4/Upstream activating sequence (UAS) system, the FLP/flippase recognition target (FRT) system, and the gonad-specific expression gene promoters (RSHP1p and Nanosp) for the germ cell-specific automatic excision of foreign DNA in the F-1 hybrid transgenic silkworms. We established 2 types of activator strains, R1p::GAL4-Gr and Nsp::GAL4-Gr, containing the testis-specific GAL4 gene expression cassettes driven by RSHP1p or Nanosp, respectively, and 1 type of effector strain, UAS::FLP-Rg, containing the UAS-linked FLP gene expression cassette. The FLP recombinase-mediated sperm-specific complete excision of FRT-flanked target DNA in the F-1 double-transgenic silkworms resulting from the hybridization of R1p::GAL4-Gr and UAS::FLP-Rg was 100%, whereas the complete excision efficiency resulting from the hybridization of Nsp::GAL4-Gr and UAS::FLP-Rg ranged from 13.73% to 80.3%. Additionally, we identified a gene, sw11114, that is expressed in both testis and ovary of Bombyx mori, and can be used to establish novel gonad-specific expression systems in transgenic silkworms. This strategy has the potential to fundamentally solve the safety issue in the production of F-1 transgenic silkworm eggs and provides an important reference for the safety of transgenic technology in other insect species.

桑树是我国重要的经济树种,属药、食同源植物.桑树白粉病对桑树有较大的危害,种类有两种,其中桑里白粉病危害最大.对桑里白粉病的研究进展进行综述,总结了桑里白粉病病原菌的鉴定和危害特征,提出了桑里白粉病抗性种质资源的识别、筛选依据,阐述了近年来桑里白粉病防控技术,针对性地提出了防控策略.对桑里白粉病未来研究、发展方向进行展望,以期为桑树病害的研究、防治与抗性育种提供参考.

家蚕丝腺拥有出色的丝蛋白合成能力,家蚕又具有容易饲养、生长周期短等优点,利用家蚕丝腺来生产大量有价值的蛋白,生产出的重组蛋白可在生物、医药、材料学等领域广泛应用,所以家蚕丝腺是一种理想的生物反应器.高效表达系统的构建是家蚕丝腺生物反应器产生重组蛋白的关键步骤之一.为了构建出更加优良的表达系统,结合国内外研究进展,对家蚕丝素重链表达系统的建立、发展与应用情况进行综述,为该系统的深入研究与应用提供参考.

Bombyx mori nuclear polyhedrosis virus（BmNPV） is one of the main pathogen in silkworm rearing. Nine kinds of BmNPV were collected and purified, then infected to the second larval silkworm of Liangguang No.2. The LC 50 of the study among 9 areas were ranged from 1.55×10~6 to 4.19×10~6 polyhedron per Milliliter, and the difference was not significant. The phylogenetic tree BmNPV of different sources based on vp39 which was amplificated, isolated and sequenced from 9 kinds of BmNPV, the results show that vp39 gene was relatively conserved. BmNPV from different sources on the evolutionary tree were grouped in the same cluster, the sequence of BmNPV from 9 areas high genetic homology which most more than 98%, and short genetic distance which most less than 1.0. The results of this study provide new clues to the antiviral mechanism of Bombyx mori.

Silkworms (Bombyx mori) reared on artificial diets during all instars have the advantages of simplicity and efficiency, year-round production, and reduced risk of poisoning. However, low silk yield remains a challenge, limiting its industrial application. To address this issue, the spinning behavior, nutrient absorption, and transcriptomics of silkworms were investigated. Compared with silkworms reared on mulberry leaves during all instars, those fed with artificial diets showed significantly lower cocoon weight, cocoon shell weight, cocoon shell rate, and silk gland tissue somatic index at the end of the fifth instar (P < 0.01). The spinning duration and crawling distance of silkworms reared on artificial diets were also significantly lower than those reared on mulberry leaves (P < 0.01). Regarding nutrient absorption, the dietary efficiency indexes of silkworms fed with artificial diets were significantly lower than those fed with mulberry leaves, except for the efficiency conversion of digesta to cocoon (P < 0.01). Further RNA-Seq analysis revealed 386 differentially transcribed genes between the 2 groups, with 242 upregulated and 144 downregulated genes. GO enrichment analysis showed that differential transcriptional genes were mainly enriched in organic acid metabolism, oxidation–reduction, and drug catabolism. KEGG enrichment analysis showed that differential transcriptional genes were mainly enriched in genetic information processing and metabolism pathways. Our findings provide new insights into the silk secretion and can serve as a reference for future research and application of silkworms fed with artificial diets.

Diapause is an important biological characteristic for many insect species to adapt to adverse environmental conditions and maintain the continuity of the race. Compared with the traditional hydrochloric acid or/and cold storage treatment methods, the artificial corona incubation technology of silkworm (Bombyx mori) eggs has many advantages including, the absence of pollution, easy operation and safety. However, this technology has not yet been applied in sericulture. In this study, we developed a novel artificial corona instrument to successfully disrupt the diapause of newly laid and refrigerated eggs from various Chinese and Japanese lineage silkworm strains. Subsequently, we invented a very early corona treatment (VECT) strategy to prevent the diapause of newly laid silkworm eggs within 4 h of oviposition. The hatching rates of the larvae were more than 95% in all diapause silkworm strains, which was comparable to the effect of the traditional HCl treatment strategy. In addition, we developed a combination strategy of VECT and pre-blastoderm microinjection and successfully created transgenic silkworms in various diapause strains. The results of the current study can aid in improving the corona artificial incubation technology and promote its application in sericulture.

Nandan white-trousers Yao people have the custom of raising local silkworm varieties Nandan white-trousers Yao silkworm to prepare flat silk. The rate of flat silk spitting, pupation, moth and others of Nandan white-trousers Yao silkworm using flat plate and folding cocooning were investigated, and the common silkworm variety Liangguang 2 was used as the control. The results shown that the spinning amount and the life rate of pupation and moth of Yao silkworm in flat cocooning were better than those in folding cocooning, while the performance of Liangguang 2 was opposite to that of Yao silkworm. Under the same condition, the flat silk spitting rate of Yao silkworm was 140.25%, which was significantly higher than that of Liangguang 2. The cocoon rate of double Palace（multi Palace） was significantly higher than that of the control variety Liangguang 2. Test confirmed that Yao silkworm is a silkworm variety suitable for spitting flat silk and was good at forming double Palace（multi Palace） cocoons.

以广西蚕区大面积饲养的二化性含多化性血统的家蚕品种两广二号一代杂交种供试,探索二化性和二化性含多化性血统的家蚕种质资源一年继代1次的蚕种保护和冷藏方法.试验首先将供试蚕种的保护分2个阶段,第1阶段将蚕种库外保护期(25℃)+库内冷藏期(3.5℃)的时间设置为320 d,第2阶段将蚕种从出库催青至化蛾产卵的时间设置为45 d,并将第1阶段又细分为9种处理:库外越夏保护时间分别为10 d、40 d、70 d、100 d、130 d、160 d、190 d、220 d、250 d,库内冷藏时间分别对应为310 d、280 d、250 d、220 d、190 d、160 d、130 d、100 d、70 d.试验结果:两广二号正交种库外越夏保护40～190 d再入库冷藏的平均孵化率均≥92％,库外越夏保护时间分别为10 d、220 d、250 d再入库冷藏的平均孵化率分别为43.20％、84.22％、65.11％;两广二号反交种库外越夏保护70～250 d再入库冷藏的平均孵化率均≥90％,均达到部颁桑蚕一代杂交种越年种孵化率≥90％的标准,而库外越夏保护时间10 d或40 d再入库冷藏的平均孵化率则只有2.52％和68.09％.以上试验结果初步提示:二化性和二化性含多化性血统的家蚕品种资源一年继代1次保种,其产卵后越夏保护时间应以不少于70 d但又不超过190 d为宜,冷藏时间以130～250 d为宜,且库外越夏保护时间与库内冷藏时间之和应该控制在320 d左右.

The Yao silkworm is a unique silkworm resource producing yellow flat plate silk that has only been reared by the Baiku Yao ethnic group in Nandan County, Guangxi Province, China for a thousand years. Here, we report the mitochondrial genomes (mitogenomes) of five Yao silkworm strains and 10 local Guangxi strains of the domestic silkworm (Bombyx mori) L. (Lepidoptera: Bombycidae), and use the resulting mitogenomes and the available Bombyx mitogenomes to characterize their genome architecture and trace the evolutionary origin of the Yao silkworm. The five Yao silkworm mitogenomes exhibited genome architectures identical to typical set of 37 mitochondrial genes (13 protein-coding genes, 22 transfer RNAs, and two ribosomal RNAs) and a high level of genome sequence similarity with the domestic silkworm. Mitogenome-based phylogenetic reconstruction provided solid evidence that the Yao silkworm shares a common ancestor with the domestic silkworm. Sliding window analysis uncovered a distinct variation pattern in the mitogenome between the Yao silkworm and the other domestic silkworm strains. The phylogenetic analyses revealed a basal placement of the Yao silkworm among all available domestic silkworm strains, indicating that the Yao silkworm is an ancient population of the domestic silkworm. Our data indicated that the Yao silkworm (B. mori) is a lineage of the domestic silkworm, which for the first time provides insights into the origin of the Yao silkworm.

实现家蚕全龄人工饲料育是未来蚕业发展的主攻方向之一.尽管已经有商业化的家蚕人工饲料,但很多家蚕品种不适宜全龄人工饲料育.本课题组通过长期的人工饲料育选择,培养出了适宜人工饲料育的家蚕品种桂蚕5号,并已经在生产上推广.为了解桂蚕5号全龄人工饲料育蚕茧缫丝后生丝品质是否与桑叶育有差异,对全龄人工饲料育家蚕生丝(SL)和桑叶育家蚕生丝(SY)进行蛋白质成分的比较分析;同时,辅以6A级生丝作为高品质生丝对照.结果表明,3个样品检测到共有蛋白242种,SL含有特有蛋白15种,SY含有13种,6A级生丝含有13种,主要为一些低丰度蛋白.对共有蛋白进行含量差异分析,结果显示,SL与SY的差异蛋白总共只有20种,SL与6A级生丝的差异蛋白有14种.对LFQ强度值排名前20的蛋白质进行分析发现,主要是丝素蛋白、丝胶蛋白、类P25蛋白及未知功能蛋白,与已经报道的结果基本一致.而且,SL的丝素蛋白、丝胶蛋白含量与SY和6A级生丝相比都没有显著差异.利用丝蛋白抗体对3种生丝的丝胶和丝素蛋白进行Western blot验证,发现丝素蛋白Fib-L和P25以及丝胶蛋白Ser1和Ser3在3种生丝中的信号强度基本一致,但在SL中检测到了强烈的Ser2蛋白信号,由于Ser2为非茧丝蛋白,推测其对生丝的性能没有大的影响.综上所述,桂蚕5号进行全龄人工饲料育所缫制生丝的蛋白质成分含量与桑叶育生丝及6A级生丝没有显著差异,具有替代桑叶育所缫制生丝用于后期高品质茧丝制品开发和应用的潜能.

广西南丹白裤瑶蚕是南丹白裤瑶族群众自繁、自育、自用的一个家蚕品种,千百年来一直相对封闭饲养,从未采取过微孢子检疫等防控措施,但也没有绝种.为探明其能保存至今的原因,采用人工添食的方法,向白裤瑶蚕2龄起蚕添食不同浓度的家蚕微孢子(Nb),并以常规家蚕品种932为对照品种同步试验,通过显微镜检测调查食下感染率,以研究瑶蚕对家蚕微孢子的耐受性.试验结果显示,家蚕微孢子浓度达到3.0×105个/mL时,对瑶蚕的感染率达到100％;瑶蚕对家蚕微孢子的食下感染率与932无显著差异,且半数感染浓度(IC50)略低于932;瑶蚕对Nb没有特殊的抗性.

[目的]本研究探讨使用人工饲料部分替代桑叶饲育后家蚕Bombyx mori幼虫肠道细菌群落的变化情况,并分析人工饲料替代桑叶的不同饲育模式下肠道关键细菌与蛹重和茧层量的相关性,为人工饲料部分替代法饲育家蚕的实际应用提供理论依据.[方法]采用全龄桑叶育(Mul1-5)、1-2龄人工饲料育+3-5龄桑叶育(Art1-2)、1-3龄人工饲料育+4-5龄桑叶育(Art1-3)、1-4龄人工饲料育+5龄桑叶育(Art1-4)和全龄人工饲料育(Art1-5)5种饲养模式饲养家蚕幼虫,统计5种饲养模式下家蚕的全茧量、茧层量及蛹重;收集不同饲养模式下家蚕5龄幼虫肠道样品,通过高通量测序技术分析其肠道菌群的组成和多样性差异,采用Spearman相关性热图分析肠道细菌与蛹重和茧层量的相关性.[结果]人工饲料与桑叶不同搭配的饲育模式对家蚕茧质有显著的影响,Mul1-5和Art1-5的全茧量最高,二者无显著性差异,Art1-2其次;Mul1-5茧层量最高,Art1-2其次,Art1-5茧层量最低.在不同饲育模式下,家蚕肠道细菌多样性和组成显著不同:在门水平上,Art1-2和Mul1-5的细菌组成最相近,优势菌主要是变形菌门(Proteobacteria)和厚壁菌门(Firmicutes),而Art1-3,Art1-4和Art1-5的细菌组成最相近,变形菌门丰度随着人工饲料饲养时间的增加逐渐增加,厚壁菌门的丰度则呈现相反的趋势;在属水平上,家蚕肠道优势菌属有假单胞菌属Pseudomonas、肠球菌属Enterococcus、鞘脂单胞菌科未分类菌属Sphingomonadaceae unclassified及罗尔斯通菌属Ralstonia等.不同饲育模式下家蚕肠道菌属丰度差异最大的是肠球菌属,Art1-2组中肠球菌属丰度最高(40.9％),随着人工饲料饲养时间的延长,丰度逐渐降低,在Art1-5中仅为0.02％.家蚕肠道菌群大部分关键物种均与茧质呈一定的相关性:柯克斯体属Coxiella与蛹重呈显著正相关,与茧层量呈显著负相关;葡萄球菌属Staphylococcus与蛹重呈显著负相关;肠球菌属与茧层量呈显著正相关.[结论]饲育模式Art1-2最接近于Mul1-5,可为人工饲料部分替代桑叶饲育法饲育家蚕提供参考.人工饲料部分替代桑叶后家蚕肠道菌群结构发生了显著变化,肠球菌属丰度的显著降低及假单胞菌属的显著增加可能与全龄人工饲料养家蚕体质弱有关;一些关键细菌与家蚕蛹重和茧层量存在显著相关性,其影响机制需要进一步研究.

桑树硬枝扦插育苗除受自身遗传条件影响外,与生根剂的处理方式、插穗本身和扦插种植的环境也密切相关.以广西壮族自治区的桑种质一年生硬枝为材料,探究生根剂、插穗、扦插期及育苗基质四大因素对扦插育苗繁殖能力的影响.结果 表明:生根剂的种类与使用方法对插穗生根有重要影响,其中以100～ 115 mg/L ABT生根粉浸药1～2h的插穗发根成苗效果最好;插穗选择当年生硬枝的中下段,规格以纵长20～25 cm、横切直径0.9～1.3 cm为宜;剪取插穗用枝的最佳时期应在一年生硬枝桑芽脱苞期和脱苞前期;不同桑种质插穗的扦插效果存在差异,其中桑种质珍珠白、日本甜橙的插穗扦插成活率最高;扦插时期宜选择在冬伐时期,以冬季至夏季(低温渐进高温)的扦插苗成活率和生长量(主茎高度、新生叶片数及根长)最高;室内试验扦插基质以雾化基质最佳,室外试验以常规园艺育苗土和细河沙为基质的插穗扦插成活率最高.研究结果将为进一步提升桑树扦插育苗技术提供借鉴.