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1. 鉴定新型反义长链非编码RNA APMAP-AS调节猪脂肪生成分化和炎症反应
ZHANG Lin-zhen, HE Li, WANG Ning, AN Jia-hua, ZHANG Gen, CHAI Jin, WU Yu-jie, DAI Chang-jiu, LI Xiao-han, LIAN Ting, LI Ming-zhou, JIN Long
Journal of Integrative Agriculture    2023, 22 (8): 2483-2499.   DOI: 10.1016/j.jia.2022.11.005
摘要208)      PDF    收藏

脂肪沉积是猪最重要的经济性状之一,是猪遗传学研究和育种工作中关注的焦点,其分子调控机制一直被视为猪分子育种重要研究内容。猪的生理结构、器官大小、代谢特征以及脂肪的生理解剖位置与人类更为相似,脂肪易沉积、易解剖和易获取,这使得猪成为研究不同内脏脂肪组织结构与功能的理想选择。长链非编码RNA (long non-coding RNA, lncRNA)是一类具有重要基因调控功能的RNA分子,在多种生物学过程中起着重要作用。随着对lncRNAs研究的深入,人们发现它不仅影响脂肪细胞的合成,还对脂肪过度沉积伴随的肥胖和代谢综合征相关的全身低度炎症具有调节作用。然而,lncRNAs对猪的内脏脂肪组织沉积过程中的调控作用还有待进行进一步研究。因此,本研究以成年雌性巴马猪(2年)为研究对象,分两组进行基础饮食和高脂饮食饲喂后,通过转录组测序技术对两组个体的内脏脂肪(腹膜后脂肪)进行测序。通过分析两组样本的测序数据,筛选了差异表达的lncRNAs,在细胞水平上进行lncRNAs的功能验证。在这里,我们在猪的基因组中发现了一个新的反义转录物,命名为APMAP-AS,由脂肪细胞膜相关蛋白(APMAP)转录而成。与对照组猪相比,APMAP-ASAPMAP在肥胖猪的腹膜后脂肪中高度表达。利用猪骨髓间质干细胞的脂肪分化模型,我们发现APMAP-AS对脂肪形成成分化有积极的调节作用。同时,APMAP-AS促进了脂质代谢并抑制了炎症因子的表达。APMAP-AS有可能与APMAP形成一个RNA-RNA双链,并增加APMAP mRNA的稳定性,从而起到正向调控猪体内APMAP表达的作用。这些结果表明,APMAP-AS通过顺式作用调节APMAP基因的表达,从而影响脂肪沉积、脂质代谢和巴马猪腹膜后脂肪组织的免疫反应。APMAP-AS是一种适应性机制,在肥胖期间促进脂肪组织的健康重塑并维持代谢平衡。这些与脂质合成相关的调控基因的天然反义转录物的发现,可能会进一步加深我们对lncRNAs在驱动适应性脂肪组织重塑和维护代谢健康方面的理解。

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2. 植物乳杆菌对藤茶副产物青贮品质影响的研究
WANG Yuan, ZHOU Hong-zhang, GAO Yu, WANG Ning-wei, LIU Han, YANG Fu-yu, NI Kui-kui
Journal of Integrative Agriculture    2023, 22 (4): 1172-1183.   DOI: 10.1016/j.jia.2022.10.001
摘要221)      PDF    收藏

本文研究目是探究青贮技术以及植物乳杆菌在藤茶副产物贮藏中的应用实验分为两组,包括对照组和植物乳杆菌处理组。在室温(23-30°C)条件下贮藏7、14和30,分析藤茶副产物青贮发酵品质、营养成分、抗氧化活性以及细菌群落。结果显示,与对照组相比,植物乳杆菌组具有较低(P<0.05)的pH和氨态氮,以及较高(P<0.05)的乳酸含量。虽然总酚在青贮过程中变化较小,但乳酸菌处理组具有较高P<0.05ABTS自由基清除能力。青贮30天后,Firmicutes取代Proteobacteria成为优势菌门,Lactobacillus成为优势属Spearman相关性结果显示乳酸与乳杆菌呈正相关关系(P<0.01)。总之,用植物乳杆菌来青贮藤渣副产物可以有效地改善青贮发酵品质和减少营养物质损失,为藤渣副产物饲料化利用提供了新的研究思路。

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3. JIA-2022-0156 PpMAPK6与PpDAM6相互作用调控桃芽内休眠解除
ZHANG Yu-zheng, XU Chen, LU Wen-li, WANG Xiao-zhe, WANG Ning, MENG Xiang-guang, FANG Yu-hui, TAN Qiu-ping, CHEN Xiu-de, FU Xi-ling, LI Ling
Journal of Integrative Agriculture    2023, 22 (1): 139-148.   DOI: 10.1016/j.jia.2022.09.010
摘要205)      PDF    收藏

休眠相关的MADS-box (DAM)基因PpDAM6在芽休眠解除过程中起关键作用,在休眠解除过程中表达降低。然而,其在调控桃花芽内休眠解除中的互作网络仍不够完善。在本研究中,我们使用酵母双杂交(Y2H)技术,桃休眠相关SSHcDNA文库中鉴定了一种丝裂原活化蛋白激酶PpMAPK6,它与PpDAM6相互作用。PpMAPK6主要位于细胞核中。进一步的Y2H和双分子荧光互补(BiFC)实验验证了PpMAPK6通过结合PpDAM6MADS-box结构域与PpDAM6相互作用。实时荧光定量PCR (qRT-PCR)分析结果表明,在3个不同需冷量的品种(春捷,中油5号,青州蜜桃)PpMAPK6的表达趋势与PpDAM6相反。此外,ABA抑制PpMAPK6在花中的表达,促进PpDAM6在花中的表达。结果表明,PpMAPK6可能通过与PpDAM6相互作用使其磷酸化,从而加速其降解。桃花内休眠解除过程中随着ABA含量的降低,PpMAPK6的表达量增加,进而降低了PpDAM6的表达量,促进桃花芽内休眠解除

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4. PPAR gamma2:正调控鸡Plin1基因表达的PPARγ主要蛋白亚型
SUN Yu-hang, ZHAI Gui-ying, PANG Yong-jia, LI Rui, LI Yu-mao, CAO Zhi-ping, WANG Ning, LI Hui, WANG Yu-xiang
Journal of Integrative Agriculture    2022, 21 (8): 2357-2371.   DOI: 10.1016/S2095-3119(21)63896-0
摘要152)      PDF    收藏

本研究的目的旨在确定鸡PPARγPlin1基因调控作用,并阐明其确切的分子机制。本研究首先利用RT-qPCR技术检测PPARγ激动剂对cPlin1基因表达的影响,而后通过双荧光素酶报告基因和RT-qPCR技术分析PPARγcPlin1基因启动子活性和mRNA表达的影响,再通过免疫共沉淀和双荧光素酶报告基因技术研究PPARγ与RXRα的协同作用对cPlin1基因启动子活性的影响,最后通过启动子截短和突变分析以及凝胶阻滞技术确定cPlin1基因启动子中PPARγ2的具体调控位点。基因表达分析结果表明,PPARγ的特异性激动剂—曲格列酮可以显著增强(P<0.05)PPARγ的靶基因LPLA-FABPFAS基因Plin1基因的mRNA表达水平,提示cPlin1基因的表达可能受PPARγ的调控;进一步的报告基因和基因表达分析结果表明,PPARγ2能够显著促进P<0.01)cPlin1基因启动子活性及mRNA表达水平,但PPARγ1却无此作用;免疫共沉淀和报告基因结果表明,PPARγ与RXRα之间存在蛋白质相互作用;与单独过表达RXRα相比,共表达PPARγ2和RXRα显著增强(P<0.01)cPlin1基因的启动子活性,但共表达PPARγ1和RXRα则没有表现出类似的现象;启动子的截短及突变分析以及凝胶阻滞结果表明,PPARγ2可以cPlin1基因启动子的-1126/-1116位点结合促进P<0.01)cPlin1基因表达。与哺乳动物相似,(i)鸡PPARγPlin1基因的转录具有正调控作用,其中PPARγ2是发挥此调控作用的主要蛋白亚型;(ii)PPARγ2是通过与cPlin1基因启动子区域的-1126/-1116位点结合来实现促进cPlin1基因表达作用的。本研究的创新性是明确了鸡PPARγPlin1基因表达的调控作用并揭示了PPARγ2调控Plin1基因转录的分子机制


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5. JIA-2021-0958 整合全基因组关联分析和选择信号分析揭示鸡F2群体骨骼肌产肉性状的遗传因子
LI Yu-dong, BAI Xue, LIU Xin , WANG Wei-jia, LI Zi-wei, WANG Ning, XIAO Fan, GAO Hai-he, GUO Huai-shun, LI Hui, WANG Shou-zhi
Journal of Integrative Agriculture    2022, 21 (7): 2065-2075.   DOI: 10.1016/S2095-3119(21)63805-4
摘要318)      PDF    收藏

本研究以东北农业大学鸡F2资源群体(NEAURP)为材料,利用 Illumina HiSeq PE150平台进行全基因组测序(26个F0个体进行10×重测序,519个F2个体进行3×重测序)。使用SAMtools进行SNP calling,BEAGLE 4.0在默认参数设置下进行基因型填充。经过质量控制和基因型填充后,共有7,890,258个SNPs用于分析。根据GRCg6a参考基因组,使用ANNOVAR软件进行SNP注释。基于混合线性模型(MLM),使用GEMMA软件进行全基因组关联分析。使用FST和π两种选择信号方法评估F2群体的遗传分化和遗传多样性。 GWAS与选择信号的整合分析表明,控制鸡骨骼肌产肉性状的遗传因子主要位于第1染色体(168.95Mb-172.43Mb)和第4染色体(74.37Mb-75.23Mb)上,共鉴定出17个可能影响目标性状的位置候选基因( LRCH1、CDADC1、CAB39L、LOC112531568、LOC112531569、FAM124A、FOXO1、NBEA、GPALPP1、RUBCNL、ARL11、KPNA3、LHFP、GBA3、LOC112532426、KCNIP4、SLIT2),其中KPNA3FOXO1是与鸡产肉性状相关的强烈候选基因。本研究的主要创新点是结合GWAS和选择信号分析方法解析鸡骨骼肌产肉性状的遗传结构,发现了一些新的影响鸡产肉性状的基因组区域和候选基因。

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6. Integration of association and computational methods reveals functional variants of LEPR gene for abdominal fat content in chickens
LI Yu-dong, WANG Wei-jia, LI Zi-wei, WANG Ning, XIAO Fan, GAO Hai-he, GUO Huai-shun, LI Hui, WANG Shou-zhi
Journal of Integrative Agriculture    2021, 20 (10): 2734-2748.   DOI: 10.1016/S2095-3119(20)63575-4
摘要149)      PDF    收藏

瘦素受体(LEPR)是瘦素的高亲和力受体,在人类和动物肥胖中起着至关重要的作用。本研究的目的是以东北农业大学肉鸡双向选择品系(NEAUHLF)为研究材料,通过关联分析和电子计算分析相结合的方法,研究LEPR外显子功能变异对鸡脂肪沉积的影响。使用5种在线生物信息学工具预测编码区单核苷酸多态性(SNPs)的功能。进一步,通过基于氨基酸残基的保守性和稳定性分析、蛋白质配体结合位点的预测、蛋白质二级结构分析、蛋白质三级结构的建模等生物信息学分析,确定了高置信度SNPs的可能结构与功能。同时,对鸡LEPR基因外显子20个非同义单核苷酸多态性(nsSNPs)与腹脂性状进行了关联分析。5种在线生物信息学工具显示,rs731962924(N867I)和rs13684622(C1002R)是最可能影响鸡腹部脂肪性状的功能性nsSNPs。氨基酸残基稳定性和保守型分析显示,大部分nsSNPs可引起蛋白质稳定性下降,rs731962924(N867I)和rs13684622(C1002R)在进化过程中相对保守。蛋白质结构分析显示rs731962924(N867I)和rs13684622(C1002R)均在可引起LEPR蛋白质结构和功能的显著变化。关联分析显示rs13684622(C1002R)与鸡腹脂重和腹脂率显著相关(P=0.0413,P=0.0260)。因此,我们认为rs13684622(C1002R)可能是一个影响鸡腹脂沉积的重要功能性SNP,有望应用于分子标记辅助选择(MAS)中培育低脂肉鸡品系。本研究的主要创新点是结合了多种生物信息学方法和nsSNPs与腹脂性状之间的关联分析进行LEPR基因功能性SNPs的筛选,为后续进行深入的功能分析提供优先研究SNP(priorization SNP)。此外,本研究用到的关联分析与计算机电子计算分析相结合的方法为鉴定农业动物重要经济性状的功能性分子标记提供了一条新的途径。


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7. Molecular characterization and functional analysis of two new lysozyme genes from soybean cyst nematode (Heterodera glycines)
WANG Ning, PENG Huan, LIU Shi-ming, HUANG Wen-kun, Ricardo Holgado, Jihong Liu-Clarke, PENG De-liang
Journal of Integrative Agriculture    2019, 18 (12): 2806-2813.   DOI: 10.1016/S2095-3119(19)62766-8
摘要111)      PDF    收藏
Soybean cyst nematode (SCN, Heterodera glycines (I.)) is one of the most important soil-borne pathogens for soybeans.  In plant parasitic nematodes, including SCN, lysozyme plays important roles in the innate defense system.  In this study, two new lysozyme genes (Hg-lys1 and Hg-lys2) from SCN were cloned and characterized.  The in situ hybridization analyses indicated that the transcripts of both Hg-lys1 and Hg-lys2 accumulated in the intestine of SCN.  The qRT-PCR analyses showed that both Hg-lys1 and Hg-lys2 were upregulated after SCN second stage juveniles (J2s) were exposed to the Gram-positive bacteria Bacillus thuringiensis, Bacillus subtilis or Staphylococcus aureus.  Knockdown of the identified lysozyme genes by in vitro RNA interference caused a significant decrease in the survival rate of SCN.  All of the obtained results indicate that lysozyme is very important in the defense system and survival of SCN. 
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8. Transcriptomic responses to aluminum (Al) stress in maize
XU Li-ming, LIU Chan, CUI Bao-ming, WANG Ning, ZHAO Zhuo, ZHOU Li-na, HUANG Kai-feng, DING Jian-zhou, DU Han-mei, JIANG Wei, ZHANG Su-zhi
Journal of Integrative Agriculture    2018, 17 (09): 1946-1958.   DOI: 10.1016/S2095-3119(17)61832-X
摘要485)      PDF    收藏
Aluminum (Al) toxicity is a major factor limiting crop production and plant growth in acid soils.  The complex inheritance of Al toxicity and tolerance mechanisms in maize has uncharacterized yet.  In this study, the maize inbred line 178 seedlings were treated with 200 μmol L–1 CaCl2+0 μmol L–1 AlCl3 (control) and 200 μmol L–1 CaCl2+60 μmol L–1 AlCl3 (Al treatment) for 1 and 6 h, respectively.  The experiment was repeated three times.  Then a detailed temporal analysis of root gene expression was performed using an Agilent GeneChip with 34 715 genes, only the genes showing more than 2.0-fold difference (P<0.01) between the control and the Al treatment maize seedlings were analyzed further.  Thus, a total of 832 different expression genes, 689 significantly up-regulated and 143 down-regulated, were identified after the seedlings were treated with Al for 6 h.  And 60 genes, 59 up-regulated and one down-regulated, were also detected after the seedlings were treated for 1 h.  Replicated transcriptome analyses further showed that about 61% of total significantly genes could be annotated based on plant genome resources.  Quantitative real-time PCR (qRT-PCT) of some selected candidate genes was used to demonstrate the microarray data, indicating significant differences between the control and Al-treated seedlings.  Exposure to Al for 6 h triggered changes in the transcript levels for several genes, which were primarily related to cell wall structure and metabolism, oxidative stress response, membrane transporters, organic acid metabolism, signaling and hormones, and transcription factors, etc.  After Al-treated for 1 h, differential abundance of transcripts for several transporters, kinase, and transcription factors were specifically induced.  In this study, the diversity of the putative functions of these genes indicates that Al stress for a short stage induced a complex transcriptome changes in maize.  These results would further help us to understand rapid and early mechanisms of Al toxicity and tolerance in maize regulated at the transcriptional level.
 
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9. Seed priming improves early seedling vigor, growth and productivity of spring maize
Hafeez ur Rehman, Hassan Iqbal, Shahzad M A Basra, Irfan Afzal, Muhammad Farooq, Abdul Wakeel, WANG Ning
Journal of Integrative Agriculture    2015, 14 (9): 1745-1754.   DOI: 10.1016/S2095-3119(14)61000-5
摘要2665)      PDF    收藏
Potential of seed priming treatments in improving the performance of early planted maize was evaluated against timely planting. Seeds of maize hybrid FH-810 were soaked in water (hydropriming), CaCl2 (2.2%, osmopriming), moringa leaf extracts (MLE 3.3%, osmopriming) and salicylic acid (SA, 50 mg L–1, hormonal priming) each for 18 h. Untreated and hydroprimed seeds were taken as control. Seeds primed with SA took less time in emergence and had high vigor in early planted maize. Amongst treatments, hormonal priming, reduced the electrical conductivity, increased the leaf relative and chlorophyll contents followed by osmopriming with CaCl2 at seedling stage. Likewise, plant height, grain rows and 1 000-grain weight, grain and biological yield and harvest index were also improved by seed priming; however hormonal priming and osmopriming with MLE were more effective in this regard. Improved yield performance by hormonal priming or osmopriming with MLE in early planting primarily owed to increased leaf area index, crop growth and net assimilation rates, and maintenance of green leaf area at maturity. In conclusion, osmopriming with MLE and hormonal priming with SA were the most economical treatments in improving productivity of early planted spring maize through stimulation of early seedling growth at low temperature.
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