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通过T-DNA插入诱变鉴定引起西瓜炭疽病的病原菌中与致病性相关的基因。

Identifying pathogenicity-related genes in the pathogen causing watermelon anthracnose disease T-DNA insertion mutagenesis.

作者信息

Guo Zhen, Wu Huijie, Peng Bin, Kang Baoshan, Liu Liming, Luo Chaoxi, Gu Qinsheng

机构信息

National Key Laboratory for Germplasm Innovation and Utilization of Horticultural Crops, Zhengzhou Fruit Research Institute, Chinese Academy of Agricultural Sciences, Zhengzhou, China.

National Key Laboratory for Germplasm Innovation and Utilization for Fruit and Vegetable Horticultural Crops, College of Plant Science and Technology, Huazhong Agricultural University, Wuhan, China.

出版信息

Front Microbiol. 2023 Jul 20;14:1220116. doi: 10.3389/fmicb.2023.1220116. eCollection 2023.

Abstract

Fruit rot caused by is a crucial watermelon disease threatening the production and quality. To understand the pathogenic mechanism of , we optimized the -mediated transformation system (ATMT) for genetic transformation of . The transformation efficiency of ATMT was an average of around 245 transformants per 100 million conidia. Southern blot analysis indicated that approximately 75% of the mutants contained a single copy of T-DNA. Pathogenicity test revealed that three mutants completely lost pathogenicity. The T-DNA integration sites (TISs) of three mutants were Identified. In mutant Cm699, the TISs were found in the intron region of the gene, which encoded a protein containing AP-2 complex subunit σ, and simultaneous gene deletions were observed. Two deleted genes encoded the transcription initiation protein SPT3 and a hypothetical protein, respectively. In mutant Cm854, the TISs were found in the 5'-flanking regions of a gene that was similar to the encoding Myosin I of (78%). In mutant Cm1078, the T-DNA was integrated into the exon regions of two adjacent genes. One was 5'-3' exoribonuclease 1 encoding gene while the other encoded a WD-repeat protein retinoblastoma binding protein 4, the homolog of the MSl1 of .

摘要

由[病原体名称未给出]引起的果实腐烂是一种严重威胁西瓜生产和品质的病害。为了解[病原体名称未给出]的致病机制,我们优化了用于[病原体名称未给出]遗传转化的[转化系统名称未给出]介导的转化系统(ATMT)。ATMT的转化效率平均为每1亿个分生孢子约产生245个转化体。Southern杂交分析表明,约75%的突变体含有单拷贝的T-DNA。致病性测试显示,三个突变体完全丧失了致病性。鉴定了三个突变体的T-DNA整合位点(TISs)。在突变体Cm699中,TISs位于一个基因的内含子区域,该基因编码一种含有AP-2复合亚基σ的蛋白质,同时观察到基因缺失。两个缺失的基因分别编码转录起始蛋白SPT3和一个假定蛋白。在突变体Cm854中,TISs位于一个与[物种名称未给出]编码肌球蛋白I的基因相似(78%)的基因的5'侧翼区域。在突变体Cm1078中,T-DNA整合到两个相邻基因的外显子区域。一个是5'-3'外切核糖核酸酶1编码基因,另一个编码WD重复蛋白视网膜母细胞瘤结合蛋白4,即[物种名称未给出]的MSl1的同源物。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/033f/10399754/fcb12f2233c1/fmicb-14-1220116-g001.jpg

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