Protein sHSP26 improves chloroplast performance under heat stress by interacting with specific chloroplast proteins in maize (Zea mays)
Protein sHSP26 improves chloroplast performance under heat stress by interacting with specific chloroplast proteins in maize (Zea mays)
复制标题
蛋白质 sHSP26 通过与玉米 (Zea mays) 中的特定叶绿体蛋白相互作用，提高热应激下叶绿体的性能
DOI：
10.1016/j.jprot.2014.12.009
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发表时间：
2015-02-06
影响因子：
3.3
通讯作者：
Wang, Wei
中科院分区：
文献类型：
--
作者：
Hu, Xiuli;Yang, Yanfang;Wang, Wei
我们最近发现，叶绿体小HSP 26（sHSP 26）是丰富的玉米叶片在热胁迫下，并可能参与玉米耐热性。然而，在很大程度上仍不清楚热应激下sHSP 26在玉米中的功能。本研究采用双向电泳技术、RNA干扰技术、免疫共沉淀技术和酵母双杂交技术等研究手段，揭示了叶绿体中与sHSP 26相互作用的蛋白质以及sHSP 26在热胁迫下的功能。在sHSP 26沉默后，分离的原生质体中共有45个蛋白质点发生了显著变化，其中33个蛋白质点为叶绿体蛋白质点。Co-IP分析显示9个蛋白可能与5 HSP 26相关。Y2 H表明，在热胁迫下，6个叶绿体蛋白与5 HSP 26相互作用。其中，ATP合成酶β亚基、叶绿素a B结合蛋白、放氧增强子蛋白1和光系统I反应中心亚基IV等4种蛋白与sHSP 26相互作用强烈，且热胁迫下5 HSP 26的RNAi后其丰度显著下降。此外，叶绿体中H2 O2的积累显著增加了sHSP 26的表达，抑制sHSP 26的表达显著降低了热胁迫下光系统II的O-2释放速率。总之，这些研究结果证明了sHSP 26在热胁迫下保护玉米叶绿体的相关性。频繁的高温胁迫会显著降低玉米的产量和品质。我们的研究结果表明，5 HSP 26通过与特定的蛋白质相互作用来改善热胁迫下玉米叶绿体的性能。这些研究结果对玉米热胁迫反应机理的理解和耐热分子育种具有重要意义。(C)2014爱思唯尔有限公司版权所有。
We recently demonstrated that chloroplast small HSP26 (sHSP26) is abundant in maize leaves under heat stress and potentially involved in maize heat tolerance. However, it largely remains unclear how sHSP26 functions in maize under heat stress. Here, 2-DE-based proteomics, RNA interference (RNAi), co-immunoprecipitation (Co-IP) and yeast two-hybrid (Y2H) were used to reveal chloroplast proteins interacting with sHSP26 and how sHSP26 functions under heat stress. After the silencing of sHSP26, a total of 45 protein spots from isolated protoplasts were greatly changed in abundance, of which 33 spots are chloroplastic. Co-IP revealed that nine proteins possibly associated with 5HSP26. Y2H demonstrated that six chloroplast proteins interact with 5HSP26 under heat stress. In particular, four proteins, including ATP synthase subunit beta, chlorophyll a b binding protein, oxygen-evolving enhancer protein 1 and photosystem I reaction center subunit IV, strongly interacted with sHSP26 and their abundance greatly declined after RNAi of 5HSP26 under heat stress. In addition, H2O2 accumulation in the chloroplasts significantly increased the expression of sHSP26, and the suppression of sHSP26 expression significantly reduced the O-2 evolution rate of photosystem II under heat stress. Overall, these findings demonstrate the relevance of sHSP26 in protecting maize chloroplasts under heat stress.Biological significanceMaize is one of the most important crops worldwide. Frequent heat stress reduces significantly the yield and quality of maize. Our results demonstrated that 5HSP26 improved maize chloroplast performance under heat stress by interacting with specific proteins. These findings are useful for understanding the mechanism of heat stress response and heat-tolerant molecular breeding in maize. (C) 2014 Elsevier B.V. All rights reserved.