程序性细胞死亡因子10抑制Ⅰ型干扰素表达并促进FMDV复制     

曾宗波  马旭升  罗志宽  齐亚银  郑海学 《畜牧兽医学报》2021,52(2):450-459

旨在探究宿主蛋白程序性细胞死亡因子10（programmed cell death factor 10，PDCD10）通过抑制Ⅰ型干扰素表达进而促进口蹄疫病毒（foot-and-mouth disease virus，FMDV）的复制。首先，本研究验证了过表达和沉默PDCD10对FMDV复制的影响，接着利用双荧光素酶报告系统探究PDCD10对Ⅰ型干扰素信号通路活化的影响，最后，利用实时荧光定量PCR探究PDCD10对Ⅰ型干扰素通路下游刺激基因（IFN-stimulated genes，ISGs）转录的影响。结果表明，过表达PDCD10显著促进FMDV的复制，沉默PDCD10显著抑制FMDV的复制。与对照相比，过表达PDCD10后感染仙台病毒（Sendai virus，SeV）的细胞培养液上清液显著促进FMDV复制，进一步，PDCD10显著抑制SeV诱导的IFN-β启动子以及NF-κB的激活且呈剂量依赖性，并且PDCD10负调控Ⅰ型干扰素通路信号分子转录，最后还发现PDCD10负调控Ⅰ型干扰素下游ISGs转录。本研究结果为深入探究PDCD10在抗病毒天然免疫中的作用积累了资料。  相似文献   

马铃薯热激转录因子HsfA3基因的克隆及其耐热性功能分析     

唐锐敏  贾小云  朱文娇  印敬明  杨清 《作物学报》2021,(4):672-683

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耐低温耐低氧萌发野败不育系赣野A的选育   总被引：1，自引：0，他引：1  

陈明亮  沈雨民  罗世友  熊焕金  吴小燕  熊文涛  肖叶青 《杂交水稻》2021,(2):25-27

赣野A是江西省农科院水稻研究所将东乡野生稻的耐冷性和耐低氧萌发能力导入抗稻瘟病保持系赣香B后再与赣香A测交和回交育成的野败型三系籼稻不育系。该不育系败育彻底,异交结实率高,配合力较强,具有较强的耐冷性和耐低氧萌发能力,在直播杂交稻育种中应用前景广阔。2019年通过了江西省品种审定。  相似文献   

超表达OsPR1A增强了Xa21介导的水稻对白叶枯病的抗性反应     

刘玉晴  燕高伟  张彤  兰金苹  郭亚璐  李莉云  刘国振  窦世娟 《中国农业科学》2021,54(23):4933-4942

【背景】前期研究发现,水稻病程相关蛋白质OsPR1A的表达受上游抗病基因Xa21调控,接菌后早期启动Xa21介导的OsPR1A较高水平表达对水稻抵抗白叶枯病菌至关重要。同时OsPR1A也受到水稻白叶枯病菌（Xanthomonas oryzae pv. oryzae,Xoo）的诱导表达。对于OsPR1A的研究绝大部分是作为抗性反应发生的标志基因佐证其他基因或途径在抗性中的作用,缺乏直接的证据证实OsPR1A本身的生物学功能。【目的】通过获得OsPR1a-OX超表达转基因植株,调查其表型及农艺性状,并明确OsPR1A蛋白质表达与抗性的关系,为鉴定OsPR1A功能提供依据。【方法】通过农杆菌介导法,将构建的OsPR1a-OX转化载体转入到水稻受体4021中,利用PCR和免疫印迹（western blot,WB）技术分别在基因水平和蛋白质水平上筛选并鉴定OsPR1A超表达阳性纯合株系。在成熟期,调查OsPR1A超表达转基因植株的表型及农艺性状（株高、穗长、分蘖数、结实率和籽粒大小等）。在31℃条件下,将生长2周的水稻幼苗TP309、4021和OsPR1A超表达转基因植株接种水稻白叶枯病菌,并在接菌0、2、4、6、8、10和12 d时测量病斑长度。在接菌0、4和6 d时,收集TP309、4021和OsPR1A超表达转基因植株的水稻叶片,提取蛋白质,利用WB技术检测OsPR1A的表达特征。【结果】构建了OsPR1a-OX转化载体,并转入到受体4021中,筛选并鉴定到2个OsPR1A超表达转基因纯合株系（#704和#709）。调查了OsPR1A超表达转基因植株在成熟期的表型及农艺性状,与对照4021相比,#704和#709的株高较矮、穗长较短、分蘖数减少、结实率降低,但籽粒稍大,可能与结实率低有关。在31℃条件下,OsPR1A超表达转基因植株的病斑长度与对照4021相比明显缩短,结果具有显著性差异（P<0.05）。在接菌0、4和6 d的材料中,超表达转基因植株#704和#709中OsPR1A始终有较高水平的表达丰度,从而提高了对白叶枯病菌的抗性。【结论】采用农杆菌介导法,获得OsPR1A超表达转基因植株;超表达OsPR1A影响到水稻的正常发育过程;超表达OsPR1A后增强了Xa21介导的水稻对白叶枯病的抗性。  相似文献   

Construction of Brucella abortus A19 VirB Promoter Deletion Strain and Study on Its Biological Characteristics     

DENG Xiaoyu  HE Jinke  YANG Qin  YI Jihai  WANG Yueli  XI Jing  WANG Zhen  CHEN Chuangfu 《中国畜牧兽医》2020,47(3):864-872

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Expression of Anti-Lipopolysaccharide Factor Isoform 3 in Chlamydomonas reinhardtii Showing High Antimicrobial Activity     

Anguo Li  Ruihao Huang  Chaogang Wang  Qunju Hu  Hui Li  Xiao Li 《Marine drugs》2021,19(5)

Antimicrobial peptides are a class of proteins with antibacterial functions. In this study, the anti-lipopolysaccharide factor isoform 3 gene (ALFPm3), encoding an antimicrobial peptide from Penaeus monodon with a super activity was expressed in Chlamydomonas reinhardtii, which would develop a microalga strain that can be used for the antimicrobial peptide production. To construct the expression cluster, namely pH2A-Pm3, the codon optimized ALFPm3 gene was fused with the ble reporter by 2A peptide and inserted into pH124 vector. The glass-bead method was performed to transform pH2A-Pm3 into C. reinhardtii CC-849. In addition to 8 μg/mL zeocin resistance selection, the C. reinhardtii transformants were further confirmed by genomic PCR and RT-PCR. Western blot analysis showed that the C. reinhardtii-derived ALFPm3 (cALFPm3) was successfully expressed in C. reinhardtii transformants and accounted for 0.35% of the total soluble protein (TSP). Furthermore, the results of antibacterial assay revealed that the cALFPm3 could significantly inhibit the growth of a variety of bacteria, including both Gram-negative bacteria and Gram-positive bacteria at a concentration of 0.77 μM. Especially, the inhibition could last longer than 24 h, which performed better than ampicillin. Hence, this study successfully developed a transgenic C. reinhardtii strain, which can produce the active ALFPm3 driven from P. monodon, providing a potential strategy to use C. reinhardtii as the cell factory to produce antimicrobial peptides.  相似文献   

Bxy‐fuca encoding α‐L‐fucosidase plays crucial roles in development and reproduction of the pathogenic pinewood nematode,Bursaphelenchus xylophilus     

Jia Tang  Ruoqing Ma  Najie Zhu  Kai Guo  Yiqing Guo  Liqun Bai  Hongshi Yu  Jiafu Hu  Xingyao Zhang 《Pest management science》2020,76(1):205-214

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miR-140-3p inhibits viability,invasion and migration of non-small-cell lung cancer A549 cells by targeting PD-L1     

ZHANG Yu-xuan  LI Chun-wei  MAO Wen-hao  ZHU Ke-yan  SHAO Yang-qian  DENG Xiao-ming 《园艺学报》2019,35(1):8-14

AIM: To explore the target relationship between microRNA-140-3p (miR-140-3p) and programmed cell death ligand 1 (PD-L1) and their effect on the viability, migration and invasion of non-small-cell lung cancer A549 cells.METHODS: RT-qPCR was used to detect the miR-140-3p expression in HLF-1, A549 and H1299 cells, and then the A549 cells with the most significant difference were selected as the subsequent research object. TargetScan software and dual-luciferase reporter assay were performed to predict and confirm the target relationship between miR-140-3p and PD-L1. RT-qPCR and Western blot were used to determine the effects of miR-140-3p mimic and inhibitor on PD-L1 expression level. MTT assay was used to detect the viability of A549 cells. Transwell assay was performed to detect the migration and invasion abilities of the A549 cells.RESULTS: miR-140-3p was significantly down-regulated in the A549 cells and H1299 cells (P<0.05). Transfection with miR-140-3p mimic decreased the expression of PD-L1 and inhibited the viability, migration and invasion of the A549 cells. Transfection with pcDNA3.0-PD-L1 reversed the inhibitory effect of miR-140-3p on the viability, migration and invasion of the A549 cells.CONCLUSION: miR-140-3p inhibits the viability, migration and invasion of A549 cells by targeting PD-L1.  相似文献   

内生菌与雷公藤细胞悬浮共培养的生理生化特性     

宋欢  宋萍  封磊  洪伟  吴承祯  王一惠  申露文 《热带作物学报》2017,(2):288-294

将2株雷公藤内生真菌Fusarium oxysporum NS33与Penicillium steckii NS6、2株内生细菌Enterobacter cloacae sub sp LG3与Serratia marcescens LY1及其组合分别与雷公藤细胞悬浮共培养,对不同培养体系内雷公藤细胞的生长及其生理生化特征进行研究。结果显示,在共培养前期,与对照相比,接种单一内生菌株提高了细胞的干重,其中菌株NS6的促生效果最明显;而在共培养后期,无论是单一内生菌还是混合内生菌均对细胞生长具有抑制作用。内生真菌和菌株组合处理的培养液p H值有明显的升高,而内生细菌LY1则明显降低了培养液的p H值,其具有产酸性。另外,当雷公藤细胞同混合菌株共培养时,培养基中总糖消耗量是最大的,而接种单独菌株时则对细胞可溶性蛋白含量具有一定的提高作用。接种内生菌会影响雷公藤细胞的POD、CAT及SOD活性,与对照相比,接种单独菌株会更加提升POD与CAT的活性,而细胞MDA含量则明显下降。  相似文献   

The M43 domain-containing metalloprotease RcMEP1 in Rhizoctonia cerealis is a pathogenicity factor during the fungus infection to wheat          下载免费PDF全文

PAN Li-jun  LU Lin  LIU Yu-ping  WEN Sheng-xian  ZHANG Zeng-yan 《农业科学学报》2020,19(8):2044-2055

Wheat(Triticum aestivum L.) is an important staple crop for global human. The necrotrophic fungus Rhizoctonia cerealis is the causal pathogen of sharp eyespot, a devastating disease of wheat. Herein, we identified RcMEP1, a zinc metalloproteaseencoding gene from R. cerealis genomic sequences, and characterized its pathogenesis function. RcMEP1 expressed at markedly-high levels during R. cerealis infection process to wheat. The predicted protein RcMEP1 comprises of 287 amino acid residues and contains a signal peptide and a M43 metalloprotease domain harboring the active site motif(HEVGHWLGLYH). The assays of Agrobacterium tumefaciens-mediated transient expression in Nicotiana benthamiana leaves indicated that RcMEP1 is an apoplastic elicitor of cell death, and that the predicted signal peptide functions and is required for secretion and cell death-induction. The purified RcMEP1 protein and its M43 domain peptide were individually able to induce plant cell death and H2 O2 accumulation, and to inhibit expression of host chitinases when infiltrated into wheat and N. benthamiana leaves, while the M43 domain-deleting peptide and negative control lacked the capacity. Moreover, compared with the control pretreatment, the purified RcMEP1 protein or its M43-domain peptide resulted in enhanced pathogenesis in the inoculated wheat, whereas the M43 domain-deleting peptide failed. These results suggest that RcMEP1 acted as an important pathogenicity factor during R. cerealis infection to wheat and that its signal peptide and M43 domain are required for the secretion and pathogenesis of RcMEP1. This study provides insights into pathogenesis role of M43 domain-containing metalloproteases during R. cerealis infection to wheat.  相似文献