Analysis of differentially expressed proteins affecting insecticidal protein content in Bt cotton under hightemperature and water deficit stress using label-free quantitation
Analysis of differentially expressed proteins affecting insecticidal protein content in Bt cotton under hightemperature and water deficit stress using label-free quantitation
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利用无标记定量分析高温和缺水胁迫下影响 Bt 棉花杀虫蛋白含量的差异表达蛋白
DOI：
10.1111/jac.12438
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发表时间：
2020
影响因子：
3.5
通讯作者：
Dehua Chen
中科院分区：
文献类型：
--
作者：
Xiang Zhang;Qiaofeng Tian;Zixu Zhao;Zhaodi Dong;Yuan Chen;Dehua Chen
筛选在高温和干旱胁迫下表达的蛋白质，以研究胁迫如何影响杀虫蛋白含量。本研究旨在为Bt棉抗虫性的维持提供理论参考。采用无标记定量方法研究了高温干旱胁迫对Bt抗虫棉品种泗抗3号差异表达蛋白的影响。在受控环境中施加两种处理[40%田间持水量和38°C（HD）和70%田间持水量和32°C（CK）]。结果表明，高温干旱胁迫显著降低棉铃杀虫蛋白含量，与对照相比，棉铃杀虫蛋白含量降低了65.2 ng/g鲜重。采用无标记定量蛋白质组学方法对差异表达蛋白质进行分析，结果显示，与对照组相比，83个蛋白质表达显著上调，104个蛋白质表达显著下调（Ratio > ±2，p值< 0.05）。此外，还检测到122种新蛋白质，未检测到167种蛋白质。对京都基因与基因组百科全书（KEGG）途径的分析表明，有14条KEGG途径受到显著影响，其中富含差异表达蛋白，有3条KEGG途径（碳水化合物消化吸收、蛋白质输出和内质网蛋白质加工）与蛋白质合成有关。在碳水化合物消化吸收途径中，淀粉水解HD的能力增强，而磷酸化己糖、果糖和葡萄糖的能力降低。在蛋白输出途径中，HD多肽的合成没有受到明显影响，但多肽进入内质网的过程受到限制。在内质网途径的蛋白质加工中，泛素介导的蛋白质水解能力增强。
Differently expression proteins under high‐temperature and drought stress were screened to investigate how stress affected the insecticidal protein content. These stresses decrease insecticidal protein, so the goal of this study was to provide a theoretical reference for maintaining the insect resistance of Bt cotton. The effect of high‐temperature and drought stress on differently expression protein in Bt cotton cultivar Sikang 3 was investigated using a label‐free quantitation method. Two treatments [40% field capacity and 38°C (HD) and 70% field capacity and 32°C (CK)] were imposed in a controlled environment. The results showed that high‐temperature and drought significantly degraded insecticidal protein content in bolls, with a decrease of 65.2 ng/g fresh weight (FW) compared to CK. The analysis of differently expression proteins with a label‐free quantitative proteomic approach showed that 83 proteins were significantly up‐regulated, but 104 proteins were significantly down‐regulated (Ratio > ±2,pvalue < 0.05) compared with CK. In addition, 122 new proteins were detected and 167 proteins were not detected. The analysis of Kyoto Encyclopedia of Genes and Genomes (KEGG) pathways showed that 14 KEGG pathways, in which the difference expression proteins were enriched, were significantly affected and three KEGG pathways (carbohydrate digestion and absorption, protein export and protein processing in endoplasmic reticulum) were related to protein synthesis. In the carbohydrate digestion and absorption pathway, the capacity of starch hydrolysis for HD increased, while the capacity to phosphorylate the hexoses, fructose and glucose decreased. In protein export pathway, the synthesis of peptide for HD was not significant affected, while the process of peptide transferred into the endoplasmic reticulum was confined. In the protein processing in endoplasmic reticulum pathway, the capacity of ubiquitin‐mediated proteolysis increased.