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两种不同基因型果实感染 后抗菌肽的生化分析

Biochemical analysis of antimicrobial peptides in two different genotypes after fruit infection by .

机构信息

Laboratório de Fisiologia e Bioquímica de Microrganismos, Centro de Biociências e Biotecnologia, Universidade Estadual do Norte Fluminense Darcy Ribeiro, Campos dos Goytacazes, RJ, Brazil.

Laboratório de Melhoramento e Genética Vegetal, Centro de Ciências e Tecnologias Agropecuárias, Universidade Estadual do Norte Fluminense Darcy Ribeiro, Campos dos Goytacazes, RJ, Brazil.

出版信息

Biosci Rep. 2019 Apr 23;39(4). doi: 10.1042/BSR20181889. Print 2019 Apr 30.

Abstract

There are several phytosanitary problems that have been causing serious damage to the crops, including anthracnose. Upon attack by certain pathogens, various protein molecules are produced, which are known as proteins related to pathogenesis (PR proteins), including antimicrobial peptides such as protease inhibitors, defensins and lipid transfer proteins (LTPs). The objective of this work is to identify antimicrobial proteins and/or peptides of two genotypes from fruits infected with The fungus was inoculated into fruits by the deposition of a spore suspension (10 conidia ml), and after 24 and 48 h intervals, the fruits were removed from the humid chamber and subjected to a protein extraction process. Protein analysis of the extracts was performed by tricine gel electrophoresis and Western blotting. The distinctive bands between genotypes in the electrophoresis profiles were subjected to mass spectrometry sequencing. Trypsin inhibition assays, reverse zymographic detection of protease inhibition and β-1,3-glucanase activity assays were also performed and extracts were also tested for their ability to inhibit the growth of fungi ' There were several low molecular weight proteins in all treated samples, and some treatments in which antimicrobial peptides such as defensin, lipid transfer protein (LTP) and protease inhibitor have been identified. It was shown that the green fruits are more responsive to infection, showing the production of antimicrobial peptides in response to injury and inoculation of the fungus, what did not occur in ripe fruits under any treatment.

摘要

有几种植物检疫问题对作物造成了严重的损害,包括炭疽病。某些病原体攻击时,会产生各种蛋白质分子,这些分子被称为与发病机制相关的蛋白质(PR 蛋白),包括抗菌肽,如蛋白酶抑制剂、防御素和脂质转移蛋白(LTP)。本工作的目的是鉴定两种基因型果实中与真菌感染相关的抗菌蛋白和/或肽。通过将孢子悬浮液(10 个分生孢子 ml)沉积在果实上,将真菌接种到果实中,在 24 和 48 小时间隔后,将果实从潮湿室中取出,并进行蛋白质提取过程。通过三氯乙酸凝胶电泳和 Western 印迹分析提取物中的蛋白质。对电泳图谱中基因型之间的特征条带进行质谱测序。还进行了胰蛋白酶抑制测定、蛋白酶抑制的反向酶谱检测和β-1,3-葡聚糖酶活性测定,并测试了提取物抑制真菌生长的能力。所有处理的样品中都有几种低分子量蛋白质,并且在一些处理中鉴定出了防御素、脂质转移蛋白(LTP)和蛋白酶抑制剂等抗菌肽。结果表明,绿色果实对感染的反应更为敏感,在受伤和接种真菌时会产生抗菌肽,而在任何处理下成熟果实都不会产生这种情况。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/151f/6481241/e615ec03d22e/bsr-39-bsr20181889-g1.jpg

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