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一个PL1家族果胶裂解酶基因是导致马铃薯普通疮痂病的5T-1的致病因子。

A PL1 family pectate lyase gene was the pathogenic factor of 5T-1 causing potato common scab.

作者信息

Zhang Cuiwen, Yang Chengde, Jin Mengjun, Feng Zhonghong, Osei Richard, Cai Fengfeng, Ma Ting, Wang Yidan

机构信息

College of Plant Protection, Gansu Agricultural University, Lanzhou, China.

Plant and Bacterial Diversity Laboratory of Gansu Province, Lanzhou, China.

出版信息

Front Microbiol. 2024 Nov 27;15:1469709. doi: 10.3389/fmicb.2024.1469709. eCollection 2024.

Abstract

Pectate lyases (PL), as important polysaccharide lyases, play an important role in the infection of host plants by pathogenic. A previous study found that the PL gene was up-regulated in the interaction between 5T-1 and potatoes. In this study, 5T-1 was used as the study object, and its gene function was investigated using bioinformatics analysis, prokaryotic expression, and CRISPR-Cas9 technology. The previous results showed that the pectate lyase gene of 5T-1 was up-regulated in the pathogenic process. In this study, the gene was cloned from the genomic DNA of 5T-1. It encoded for a 415-residue protein with a complete PL-6 superfamily domain and Pec_lyase_C domain, which belongs to the PL1 family. The soluble protein encoded by was obtained successfully, which has high pathogenicity after inoculating healthy potatoes. The mutant strain △PL5T-1 with gene deletion was successfully obtained, and its colony morphology and pigment were not significantly different from that of wild strains, but its growth rate was slowed down, moreover, the hyaline circle formed by the mutant strain ΔPL5T-1 using pectin was significantly smaller than wild strain, and the deletion of this gene affected the infestation rate of 5T-1. Our results confirm that the gene was the pathogenic factors and played a key role in process of infecting and causing potato common scab, which laid foundation for understanding the pathogenic mechanism of 5T-1.

摘要

果胶酸裂解酶(PL)作为重要的多糖裂解酶,在病原菌侵染寄主植物过程中发挥着重要作用。先前的一项研究发现,PL基因在5T - 1与马铃薯的互作中上调表达。在本研究中,以5T - 1为研究对象,运用生物信息学分析、原核表达及CRISPR - Cas9技术对其基因功能进行了研究。先前的结果表明,5T - 1的果胶酸裂解酶基因在致病过程中上调表达。在本研究中,从5T - 1的基因组DNA中克隆了该基因。它编码一个含有完整PL - 6超家族结构域和Pec_lyase_C结构域的415个氨基酸残基的蛋白质,属于PL1家族。成功获得了该基因编码的可溶性蛋白,接种健康马铃薯后具有高致病性。成功获得了缺失该基因的突变菌株△PL5T - 1,其菌落形态和色素与野生菌株无显著差异,但其生长速率减慢,此外,突变菌株ΔPL5T - 1利用果胶形成的透明圈明显小于野生菌株,该基因的缺失影响了5T - 1的侵染率。我们的结果证实该基因是致病因子,在侵染和引起马铃薯疮痂病过程中起关键作用,为理解5T - 1的致病机制奠定了基础。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/d7b2/11631876/90161fb9146a/fmicb-15-1469709-g001.jpg

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