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玉米在蛋鸡饲粮中有效能值的测定及其净能预测模型的构建

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动物营养学报 2024年第4期36卷 2344-2355页

作者：潘敏慧李成云张海军班志彬陈思淼梁浩闫晓刚赵晓东赵敬慧延边大学农学院延吉133002 中国农业科学院饲料研究所北京100081 吉林省农业科学院动物营养与饲料研究所公主岭136100 吉林大北农农牧科技有限责任公司长春130000

本试验采用间接测热法结合套算法测定不同来源玉米在产蛋高峰期海兰褐蛋鸡饲粮中的有效能值并建立基于理化指标的玉米净能(NE)预测模型。选取54周龄体重为(1898.32±114.12)g、产蛋率为(90.74±1.31)%的海兰褐蛋鸡共72只,随机分为6个组(... 详细信息

本试验采用间接测热法结合套算法测定不同来源玉米在产蛋高峰期海兰褐蛋鸡饲粮中的有效能值并建立基于理化指标的玉米净能(NE)预测模型。选取54周龄体重为(1898.32±114.12)g、产蛋率为(90.74±1.31)%的海兰褐蛋鸡共72只,随机分为6个组(1个玉米-豆粕型基础饲粮组和5个玉米替代比例为30%的试验饲粮组),每组6个重复,每个重复2只鸡。试验鸡在舍内笼养,预试期10 d,正试期12 d,其中正试期分为3期,每期4 d(适应1 d,呼吸测热3 d)。每期试验中,从每组中选择4只试验鸡,称重后分别放入呼吸测热装置的12个代谢室(每个代谢室2只),每2个代谢室对应l种饲粮,间接测热和消化代谢试验同时进行。结果显示:1)5种玉米的表观代谢能(AME)分别为15.09、14.18、14.67、14.33和14.28 MJ/kg DM,NE分别为10.90、10.33、10.82、10.47和10.44 MJ/kg DM,NE/AME分别为72.17%、72.84%、73.71%、72.99%和73.11%。2)玉米的NE预测方程为NE=-0.001+0.57×AME+0.606×粗脂肪+0.088×粗纤维(R 2=0.987,P<0.01)。交叉验证显示本试验所得蛋鸡玉米的NE预测模型准确性较高,利用该模型可根据AME、粗脂肪和粗纤维含量预测玉米的NE。

关键词：蛋鸡净能间接测热法预测模型

Construction of a One-Vector Multiplex CRISPR/Cas9 Editing System to Inhibit Nucleopolyhedrovirus Replication in Silkworms

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Virologica Sinica 2019年第4期34卷 444-453页

作者： Zhanqi Dong Qi Qin Zhigang Hu Peng Chen Liang Huang Xinling Zhang Ting Tian Cheng Lu minhui pan State Key Laboratory of Silkworm Genome Biology Southwest UniversityChongqing 400716China Key Laboratory of Sericultural Biology and Genetic Breeding Ministry of AgricultureSouthwest UniversityChongqing 400716China

Recently the developed single guide(sg)RNA-guided clustered regularly interspaced short palindromic repeats/associated protein 9 nuclease(CRISPR/Cas9) technology has opened a new avenue for antiviral therapy. The CRISPR/Cas9 system uniquely allows targeting of multiple genome sites simultaneously. However, there are relatively few applications of CRISPR/Cas9 multigene editing to target insect viruses. To address the need for sustained delivery of a multiplex CRISPR/Cas9-based genome-editing vehicle against insect viruses, we developed a one-vector(pSL1180-Cas9-U6-sgRNA) system that expresses multiple sgRNA and Cas9 protein to excise Bombyx mori nucleopolyhedrovirus(BmNPV) in insect *** screened the immediate-early-1 gene(ie-1), the major envelope glycoprotein gene(gp64), and the late expression factor gene(lef-11), and identified multiple sgRNA editing sites through flow cytometry and viral DNA replication analysis. In addition, we constructed a multiplex editing vector(PSL1180-Cas9-sgIE1-sgLEF11-sgGP64, sgMultiple) to efficiently regulate multiplex gene-editing and inhibit BmNPV replication after viral infection. This is the first report of the application of a multiplex CRISPR/Cas9 system to inhibit insect virus replication. This multiplex system can significantly enhance the potential of CRISPR/Cas9-based multiplex genome engineering in insect virus.

关键词： Bombyx mori Bombyx mori nucleopolyhedrovirus(BmNPV) Antiviral therapeutic CRISPR/Cas9 Multigene editing

Boron promoted Fe^(3+)/peracetic acid process for sulfamethazine degradation:Efficiency,role of boron,and identification of the reactive species

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Journal of Environmental Sciences 2024年第1期135卷 72-85页

作者： Yanhong Zhang Guangbing Liu Yuzhu Xue Lichun Fu Yawei Qian minhui Hou Xiang Li Chen Ling Ying Zhang Yuwei pan Jiangsu Provincial Key Laboratory of Environmental Engineering Jiangsu Provincial Academy of Environmental ScienceNanjing 210036China College of Biology and the Environment Nanjing Forestry UniversityNanjing 210037China School of Iron and Steel Soochow UniversitySuzhou 215000China School of the Environment Nanjing UniversityNanjing 210023China SUMEC Complete Equipment&Engineering Co. LTD.Nanjing 211500China Key Laboratory of Pollution Process and Environmental Criteria Ministry of EducationCollege of Environmental Science and EngineeringNankai UniversityTianjin 300350China School of Environment Key Laboratory for Yellow River and Huai River Water Environmental and Pollution ControlMinistry of EducationHenan Key Laboratory for Environmental Pollution ControlHenan Normal UniversityXinxiang 453007China

In this work,boron(B)was used to promote Fe^(3+)/peracetic acid(Fe^(3+)/PAA)for the degradation of sulfamethazine(SMT).An SMT degradation efficiency of 9.1%was observed in the Fe^(3+)/PAA system over 60 min,which was significantly increased to 99.3%in the B/Fe^(3+)/PAA system over 10 *** B/Fe^(3+)/PAA process also exhibited superior resistance to natural substances,excellent adaptability to different harmful substances,and good removal of antibiotics in natural fresh water *** mechanism of action of boron for Fe^(3+)reduction was determined using scanning electron microscopy(SEM),X-ray photoelectron spectroscopy(XPS),Fourier transform infrared(FT-IR)spectroscopy,density functional theory(DFT)calculations,and electrochemical *** dominant role of^(·)OH was confirmed using quenching experiments,electron spin resonance(EPR)spectroscopy,and quantitative *** radicals(R-O^(·))and Fe(IV)also significantly contribute to the removal of *** calculations on the reaction between Fe^(2+)and the PAA were conducted to further determine the contribution from ^(·)OH,R-O^(·),and Fe(IV)from the perspective of thermodynamics and the reaction *** boron dosages,Fe^(3+)dosages,and initial pH values were also investigated in the B/Fe^(3+)/PAA system to study their effect of SMT removal and the production of the reactive ***(IV)production determined the k_(R-O·+Fe(IV))value suggesting that Fe(IV)may play a more important role than R-O^(·).A comparison of the results with other processes has also proved that the procedure described in this study(B/Fe^(3+)/PAA)is an effective method for the degradation of antibiotics.

关键词： Boron Peracetic acid Sulfamethazine DFT Reactive species

BmCyclin B and BmCyclin B3 are required for cell cycle progression of silkworm,Bombyx mori

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BmCyclin B and BmCyclin B3 are required for cell cycle progr...

第十届家（柞）蚕遗传育种及良种繁育学术研讨会

作者： pan minhui HONG KaiLi CHEN XiangYun pan Chun CHEN XueMei KUANG XiuXiu LU Cheng State Key Laboratory of Silkworm Genome Biology Southwest University

Cyclin B is an important regulator of the G2 to M phase in the cell cycle. The silkworm genomic database showed that there are two Cyclin B genes in the silkworm（Bombyx mori）, BmCyclin B and BmCyclin B3. Using silkworm ESTs data, the BmCyclin B3（EU074796） gene was cloned. Its complete cDNA length was 1,665 bp and the ORF was 1,536 bp with six introns and seven exons. The BmCyclin B3 gene encoded 511 amino acids, and the predicted molecular weight was 57.8 kD with an isoelectric point of 9.18. It consisted of one protein damage box and two cyclin boxes. Cell cycle arrest in G2 or M phase in BmN-SWU1 cells could be induced by RNA interference-mediated reduction of BmCyclin B and BmCyclin B3 expression, thus inhibiting the cell proliferation. These results suggest that BmCyclin B and BmCyclin B3 are necessary for completing the cell cycle in silkworm cells.

关键词： Bombyx mori cell cycle BmCyclin B BmCyclin B3 cell proliferation

RNAi-mediated silencing of transcription factor SoxE gene expression in the silkworm ovary-derived BmN4 cells inhibits cell proliferation

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RNAi-mediated silencing of transcription factor SoxE gene ex...

中国遗传学会第九次全国会员代表大会暨学术研讨会

作者：魏玲 Zhiqing Li Daojun Cheng Takahiro Kusakabe minhui pan Zhonghuai Xiang 鲁成 State Key Laboratory of Silkworm Genome Biology Southwest Univcrsity School of Life Science Southwest University Laboratory of Silkworm Science Kyushu University Graduate School of Biorcsource and State Key Laboratory of Silkworm Genome Biology Southwest University

Transcription factor SoxE is mainly expressed in gonad and involved in the regulation of gonad development and sex determination in *** based on silkworm ovary-derived cell line(BmN4-SID1) to survey the roles of BmSoxE on cell proliferation and predict the potential targets for ***-mediated silencing of BmSoxE expression in BmN4-SIDI cells led to a decreased cell proliferation,which is likely resulted from a cell cycle arrest at the G1 phase after BmSoxE RNAi.A further genome-wide microarray screening revealed a significant expression change of a set of genes after BmSoxE *** than half of these genes contain conserved binding motifs for HMG-box of Sox protein and were expected as the targets for *** sets of BmSoxE target candidates are associated with cell proliferation and show gonad-specific *** data will provide fundamental knowledge for investigating further the roles of BmSoxE in the control of cell cycle and silkworm development.

关键词： Bombyx mori SoxE protein RNA interference Microarray

RNAi-mediated silencing of transcription factor SoxE gene expression in the silkworm ovary-derived BmN4 cells inhibits cell proliferation

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RNAi-mediated silencing of transcription factor SoxE gene ex...

第十届家（柞）蚕遗传育种及良种繁育学术研讨会

作者： Ling Wei Zhiqing Li Daojun Cheng Takahiro Kusakabe minhui pan Zhonghuai Xiang Cheng Lu State Key Laboratory of Silkworm Genome Biology Southwest University School of Life Science Southwest University Laboratory of Silkworm Science Kyushu University Graduate School of Bioresource

Transcription factor SoxE is mainly expressed in gonad and involved in the regulation of gonad development and sex determination in animals. We based on silkworm ovary-derived cell line（BmN4-SID1） to survey the roles of BmSoxE on cell proliferation and predict the potential targets for BmSoxE. RNAi-mediated silencing of BmSoxE expression in BmN4-SID1 cells led to a decreased cell proliferation, which is likely resulted from a cell cycle arrest at the G1 phase after BmSoxE RNAi. A further genome-wide microarray screening revealed a significant expression change of a set of genes after BmSoxE RNAi. More than half of these genes contain conserved binding motifs for HMG-box of Sox protein and were expected as the targets for BmSoxE. Several sets of BmSoxE target candidates are associated with cell proliferation and show gonad-specific expression. These data will provide fundamental knowledge for investigating further the roles of BmSoxE in the control of cell cycle and silkworm development.

关键词： Bombyx mori SoxE protein RNA interference Microarray

对家蚕抗核型多角体病毒的研究策略及研究进展

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蚕业科学 2017年第1期43卷 139-146页

作者：董战旗潘敏慧家蚕基因组生物学国家重点实验室重庆400716 农业部家蚕功能基因组和生物学重点实验室西南大学重庆400716

由家蚕核型多角体病毒(Bm NPV)引发的家蚕血液型脓病是生产上危害最为严重的蚕病之一。国内外学者从研究家蚕的抗病遗传规律、选育对病毒具有抵抗力的家蚕品种,一直到应用现代分子生物学技术研究病毒的基因组、转录组和蛋白组,研究病毒... 详细信息

由家蚕核型多角体病毒(Bm NPV)引发的家蚕血液型脓病是生产上危害最为严重的蚕病之一。国内外学者从研究家蚕的抗病遗传规律、选育对病毒具有抵抗力的家蚕品种,一直到应用现代分子生物学技术研究病毒的基因组、转录组和蛋白组,研究病毒的侵染规律及与蚕体的互作,基于RNAi和CRISPR/Cas9技术培育转基因抗病毒素材等,已经取得了诸多进展。本文系统总结近10年来有关家蚕抗Bm NPV研究在上述各个方面取得的重要成果,并对未来家蚕抗病毒机制研究和抗病毒分子育种趋势以及亟需解决的科学与技术问题进行探讨,以期为不断深入的家蚕抗病毒研究提供参考。

关键词：家蚕家蚕核型多角体病毒抗性分子机制抗性育种素材

DNA Synthesis during Endomitosis Is Stimulated by Insulin via the PI3K/Akt and TOR Signaling Pathways in the Silk Gland Cells of Bombyx mori

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DNA Synthesis during Endomitosis Is Stimulated by Insulin vi...

第十二届家（柞）蚕遗传育种暨良种繁育学术研讨会

作者： Yaofeng Li Xiangyun Chen Xiaofang Tang Chundong Zhang La Wang Peng Chen minhui pan Cheng Lu State Key Laboratory of Silkworm Genome Biology Southwest University Department of Biochemistry and Molecular Biology Chongqing Medical University Key Laboratory for Sericulture Functional Genomics and Biotechnology of Agricultural Ministry Southwest University

Silk gland cells undergo multiple endomitotic cell cycles during silkworm larval ontogeny. Our previous study demonstrated that feeding is required for continued endomitosis in the silk gland cells of silkworm larvae. Furthermore, the insulin signaling pathway is closely related to nutritional signals. To investigate whether the insulin signaling pathway is involved in endomitosis in silk gland cells, in this study, we initially analyzed the effects of bovine insulin on DNA synthesis in endomitotic silk gland cells using 5-bromo-2’-deoxyuridine（Brd U） labeling technology, and found that bovine insulin can stimulate DNA synthesis. Insulin signal transduction is mainly mediated via phosphoinositide 3-kinase（PI3K）/Akt, the target of rapamycin（TOR） and the extracellular signal-regulated kinase（ERK） pathways in vertebrates. We ascertained that these three pathways are involved in DNA synthesis in endomitotic silk gland cells using specific inhibitors against each pathway. Moreover, we investigated whether these three pathways are involved in insulin-stimulated DNA synthesis in endomitotic silk gland cells, and found that the PI3K/Akt and TOR pathways, but not the ERK pathway, are involved in this process. These results provide an important theoretical foundation for the further investigations of the mechanism underlying efficient endomitosis in silk gland cells.

关键词： Bombyx mori silk gland DNA synthesis insulin signaling

RNAi silencing of the SoxE gene suppresses cell proliferation in silkworm BmN4 cells

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RNAi silencing of the SoxE gene suppresses cell proliferatio...

第十二届家（柞）蚕遗传育种暨良种繁育学术研讨会

作者： Ling Wei Zhiqing Li Daojun Cheng Takahiro Kusakabe minhui pan Jun Duan Yonghu Wang Cheng Lu State Key Laboratory of Silkworm Genome Biology Southwest University School of Life Science Southwest University Laboratory of Silkworm Science Kyushu University Graduate School of Bioresource and Bioenvironmental Sciences

The transcription factor SoxE is mainly expressed in the gonad and involved in the regulation of gonad development and sex determination in animals. Here, we used the silkworm ovary-derived BmN4-SID1 cell line to survey the roles of the silkworm SoxE protein（BmSoxE） and predict its candidate binding targets. RNAi-mediated silencing of BmSoxE expression suppressed cell proliferation in BmN4-SID1 cells. A further cell cycle analysis revealed that this inhibition of cell proliferation was largely due to cell cycle arrest in G1 phase when BmSox E expression was blocked in BmN4-SID1 cells. Genome-wide microarray expression analyses demonstrated that the expression levels of a set of genes were significantly altered following BmSox E RNAi. More than half of these genes contained conserved binding sites for HMG box domain of the Sox proteins and were predicted to be candidate binding targets for BmSoxE. Importantly, some of the candidate targets may be associated with the effect of BmSox E on cell proliferation. Several candidate target genes showed gonad-specific expression in silkworm larvae. Taken together, these data demonstrate that BmSoxE is required for cell proliferation in silkworm BmN4-SID1 cells and provide valuable information for further investigations of the molecular control exerted by the BmSoxE protein over cell proliferation and gonad development in the silkworm.

关键词： Silkworm SoxE RNAi Cell proliferation Target