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.

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 neuropeptide Orcokinin (abbreviated as BommoOK) is equipped with multiple biological functions, one of which acts as a pigmentation inhibitor. To explore the whitening efficiency of BommoOK, the inhibitory effects on tyrosinase and its adaptability on the cell for six mature peptides of BommoOK were investigated in this paper. At the same time, BommoOKA_type4, the peptide with the best melanin inhibition effect, was used as an additive to prepare a whitening cream, and the effects on skin moisture, oil content, fine lines, skin glossiness, pores, and pigment depth were determined. The results revealed that the cream added with BommoOKA_type4 peptide showed a good improvement effect on the skin, especially in significantly reducing the pigment depths of skin. The results displayed a potential application of BommoOK in whitening and skincare products as an excellent additive and provide certain references for the mechanism research of BommoOK in inhibiting melanin synthesis.

Bombyx mori is an oligophagous economic insect. Cis-Jasmone is one of the main substances in mulberry leaf that attract silkworm for feeding and BmOR56 is its receptor. Potential interaction ways between BmOR56 and cis-Jasmone were explored, which included some crucial amino acids such as Gln172, Val173, Ser176, Lys182, His322, and Arg345. BmOR56 was edited using CRISPR/cas9 for Qiufeng, and a homozygous knockout strain QiufengM was obtained. Compared with Qiufeng, the feeding ability of QiufengM on mulberry leaf did not change significantly, but on artificial diet decreased significantly. QiufengM also showed a dependence on the concentration of mulberry leaf powder. The result indicated that other olfactory genes had a compensatory effect on the attractance of mulberry leaf after the loss of BmOR56. Transcriptome analysis of antennae showed that many genes differentially expressed between Qiufeng and QiufengM, which involved in olfactory system, glucose metabolism, protein metabolism, amino acid metabolism, and insect hormone biosynthesis. Particularly, BmIR21, BmOR53 and BmOR27 were significantly up-regulated, which may have a compensatory effect on BmOR56 loss. In addition, detoxification mechanism was activated and may cause the passivation of feeling external signals in silkworm.

[Objective]By using BSA-Seq sequencing analysis to determine the resistance SNP markers and resistance genes of the resistant parent NC99R of the nuclear polyhedrosis resistant variety Guican N2,lay a foundation for the iden-tification of NC99R resistance genes and provide marker resources for silkworm nuclear polyhedrosis resistant molecular marker assisted breeding.[Method]A hybrid F1,F2,and BC1 genetic population were prepared using the disease resistant variety NC99R and the susceptible variety Furong as parents,and their genetic patterns were analyzed by feeding nuclear polyhedrosis virus(BmNPV).In the F2 population of NC99R near allelic gene line NC99R-NIL hybridized with Furong,high and low concentration feeding groups with virus concentration differences of 4000 times were set up.Non-diseased individuals were selected from the high concentration feeding group to construct an extreme resistance material DNA mixing pool.Individuals with typical disease symptoms were selected from the low concentration feeding group to con-struct an extreme susceptibility material DNA mixing pool.Then the resistance DNA mixing pool,susceptible DNA mixed pool and two parental genomes were completed sequencing using second-generation sequencing,and locating resistance linked regions with population segregation analysis(BSA).[Result]The results showed that NC99R exhibited stable high resistance to BmNPV,and the genome carried anti BmNPV genes,which were jointly regulated by significant resistance major genes or closely linked genes.A total of 19276670 SNP loci and 4789615 InDel loci were obtained through whole genome sequencing analysis of extreme resistance DNA mixing pool,extreme susceptibility DNA mixing pool and paren-tal DNA.Based on the LOESS regression fitting analysis of ΔSNP-index,two regions associated with resistance to nu-clear polyhedrosis were identified,Chr.25:150-11069kb and Chr.27:8700-11009kb respectively.When the ΔSNP-index was set to≥0.8,a total of 3865 SNP markers significantly related to resistance were identified.Among them,78 SNP markers were screened in the association interval of chromosome 25 and 2694 SNP markers were screened in the associa-tion region of chromosome 27.After screening for non-synonymous mutations at the amino acid level,the Zinc carboxy-peptidase gene on chromosome 25 and the Facilitated trehalose transporter Tret1 gene on chromosome 27 were identified.[Conclusion]Using BSA-Seq,3865 SNP marker resources related to resistance against nuclear polyhedrosis are found in silkworm NC99R,and two genes related to resistance against BmNPV are preliminarily identified(Zinc carboxypepti-dase gene on chromosome 25 and Facilitated trehalose transporter Tret1 gene on chromosome 27),providing marker re-sources for molecular marker assisted breeding of silkworms resistant to nuclear polyhedrosis.

石灰是养蚕生产中常用的消毒剂,为了解石灰消毒的几个重要性能,对石灰的有效性、稳定性、吸湿性和腐蚀性进行了深入研究.结果表明,石灰对家蚕核型多角体病毒和败血病病菌具有较好杀灭效果,对家蚕白僵菌生长也具有一定抑制效果,对家蚕微孢子虫没有杀灭效果,养蚕生产中应用石灰消毒效果表现出高效性.熟石灰粉吸湿性强,在空气中暴露、无任何保存措施的条件下21 d失效,在干燥、密封条件下稳定性好.石灰对铁质器具的腐蚀率低,属于无腐蚀性级别.石灰与其他消毒剂混合后有效成分稳定性好,是一种优质载体.综上研究表明,石灰是一种优质蚕药,养蚕生产合理使用石灰可以实现高效防病目的.

广西壮族自治区科技厅公布2023年度新认定的23家自治区重点实验室名单(桂科基字[2023]33号)中,广西壮族自治区蚕业技术推广站主持申报的"广西蚕桑遗传改良与高效种养重点实验室(Guangxi Key Laboratory of Silkworm Genetic Improvement and Efficient Breeding)"成功获批,这是广西蚕业技术推广站首个获批的自治区级重点实验室,将对广西蚕桑产业基础和应用基础研究产生巨大的推动作用,促进产业高质量可持续发展.

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.

2023年1月6日上午,"广西蚕桑遗传改良与技术创新中心实验室"实验大楼正式揭牌启用.自治区农业农村厅总农艺师李如平、自治区农业农村厅科教处处长左明、自治区蚕业技术推广站站长李标为实验室揭牌.实验大楼的正式启用对推进蚕桑科技创新体系建设、促进产业高质量发展具有重要意义.

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.

为了充分挖掘现有桑树品种的药用价值,对生产上现行推广的杂交桑品种进行同品种不同方式剪伐后,采用分光光度比色法对不同品种间和组织部位间总黄酮含量进行测定与分析.结果显示,桑树枝与叶、芽总黄酮含量差异极显著,叶和芽总黄酮含量差异不显著,以桑芽含量最高,叶次之,枝含量最低;剪伐方式对桑树总黄酮含量影响不显著;桑树不同品种间总黄酮含量不同,桑枝总黄酮含量以桂桑5号和桂桑优12最高,桑叶总黄酮含量以桂桑5号、桂桑6号最高,桑芽总黄酮含量以桂桑6号、桂桑优12最高;同一枝条不同部位的枝、叶含量也不同,其中绿枝总黄酮含量显著高于硬枝,上部叶含量高于中下部叶,中部叶高于下部叶,上部叶与下部叶差异性极显著,建议选择桑树总黄酮提取和食用组织部位时以桑芽最佳,其次是上、中部叶片.

家蚕(Bombyx mori)2龄不眠蚕(non-molting in the 2nd instar,nm2)突变体是从家蚕品种C603中发现的一种致死突变体,发育至2龄将眠期后不能正常入眠蜕皮,持续5～7 d后陆续死亡,图位克隆结果表明该突变体是由表皮蛋白BmCPG10突变引起的,且血淋巴中的蜕皮激素滴度显著低于野生型.蜕皮激素主要在前胸腺合成,因此本研究以nm2突变体和野生型家蚕2龄将眠蚕的前胸腺(包含第一对气管丛)为材料进行双向电泳分析,从中检测到一个表皮蛋白BmCPR41在突变体中的表达量高于野生型,组织表达谱表明BmCPR41在前胸腺的表达量远高于其他组织.通过RNAi敲低BmCPR41的表达,发现有部分家蚕推迟入眠或不入眠,蚕体重也显著低于对照组,且相关基因基于RNA-seq的表达情况与 nm2突变体中的基本一致.基于上述实验结果推测BmCPR41基因在nm2突变体形成,尤其蜕皮激素信号通路上发挥重要作用.本研究结果为进一步研究nm2突变体的形成机制提供了实验证据,并可为研究蜕皮激素介导的信号通路提供理论依据.

[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.

Pyriproxyfen is a juvenile hormone-like pesticide. Once intake occurs, it leads to a series of poisoning characters consequences in silkworm, Bombyx mori (ID: 7091, Lepidoptera), such as non- cocooning, non-pupation, production of low-active eggs, and extended stages. However, the poisoning mechanism is still unclear. Here, silkworms were fed mulberry leaves soaked with different pyriproxyfen concentrations, and the heads were dissected for transcriptome analysis, while the hemolymph was used for determinations of ecdysone and juvenile hormone titers. As a result, after conjoint analysis of 3 feeding groups and a control group, 555 differentially expressed genes (DEGs) were obtained, which were mainly involved in hormone metabolism, glycometabolism and protein metabolism. Meanwhile, 119 genes were significantly correlated with the pyriproxyfen concentrations, and they were mainly involved in drug metabolism and glycometabolism. The ecdysone titers in several feeding groups were significantly lower than those of the control group, while juvenile hormone was not detected in all groups, including the control and feeding groups. Correspondingly, due to activation of the juvenile hormone signaling pathway by pyriproxyfen, key genes in the ecdysone synthesis pathway were downregulated, and a large number of downstream genes were up- or downregulated. In addition, nearly all genes in the detoxification pathway were upregulated. These results suggested that, affected by the juvenile hormone signaling pathway, ecdysone titers decreased and further affected a series of downstream processes, and this was the key reason for pyriproxyfen poisoning in silkworm, B. mori, which could lay a foundation for the study of pyriproxyfen resistance in silkworm.

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.

为了探索家蚕彩色茧品种在制作平面丝中的应用潜能,研究广西区内特有家蚕彩色茧品种在高温环境下所吐平面丝的性状.结果表明,在28～32℃高温环境条件下3种家蚕彩色茧(黄色、红色和绿色)品种吐出率为67％～82％,平面丝厚薄较均匀,其中以家蚕红色茧品种红日7532·红日的平面丝性状最佳,表现为吐出率较高,达到81.94％,体质强健,没有病死蚕,在平面用具上分布均匀、不打堆、无落地蚕、无包埋蚕等,适合用于省力化制作平面丝.

Yun7Ge is a giant egg mutant found in the silkworm variety Yun7. In comparison with the giant mutant Ge, the eggs of Yun7Ge are larger. The number of laid eggs and hatching rate of Yun7Ge are reduced, which is not conducive to reproduction. In this work, the target gene controlling giant egg trait is located on the Z chromosome and was determined through genetic analysis. Transcriptome results showed that phytanoyl-CoA dioxygenase domain-containing protein 1 (PHYHD1) on the Z chromosome was silenced, and the 25 chorion genes on chromosome 2 were remarkably downregulated. Sequence analysis showed that the 73.5 kb sequence including the PHYHD1 was replaced by a ~3.0 kb sequence. After knocking out the PHYHD1 by using CRISPR/Cas9, the chorion genes were significantly downregulated. Hence, the silencing of PHYHD1 leads to the downregulation of many chorion protein genes, thus directly causing giant eggs.

插穗是扦插苗生命营养的直接供给体,是影响扦插育苗质量的最客观因素.从插穗用枝的萌芽状态、用枝部位、规格及不同生长时期等方面进行试验研究.结果显示,桑树插穗用枝要选择在未萌芽期,选择枝条的中、下段部分为宜,插穗规格因不同品种存在差异,以插穗长10～20 cm,横切面直径1.0～1.5 cm时的育苗质量较好.生产上常规的果桑品种中,以珍珠白、日本甜橙及金樯63号插穗的生根成活率表现更好,成活率均达94％以上.冬伐枝插穗育苗显著好于夏伐枝插穗苗,除考虑季节影响因素外,对插穗枝的活性成分进行了进一步的研究,发现冬季桑枝的活性成分都高于夏季枝.一元线性相关性分析得出多糖、总生物碱的含量与苗木的成活率呈高度相关.