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蛋白质组学分析揭示了PEG 胁迫下敏感和抗旱高粱响应的不同分子机制。

Proteomic Analysis Revealed Different Molecular Mechanisms of Response to PEG Stress in Drought-Sensitive and Drought-Resistant Sorghums.

机构信息

Biotechnology Research Institute, Chinese Academy of Agricultural Sciences, Beijing 100081, China.

Institute of Crop Sciences, Chinese Academy of Agricultural Sciences, Beijing 100081, China.

出版信息

Int J Mol Sci. 2022 Oct 31;23(21):13297. doi: 10.3390/ijms232113297.

Abstract

Drought is the major limiting factor that directly or indirectly inhibits the growth and reduces the productivity of sorghum ( (L.) Moench). As the main vegetative organ of sorghum, the response mechanism of the leaf to drought stress at the proteomic level has not been clarified. In the present study, nano-scale liquid chromatography mass spectrometry (nano-LC-MS/MS) technology was used to compare the changes in the protein expression profile of the leaves of drought-sensitive (S4 and S4-1) and drought-resistant (T33 and T14) sorghum varieties at the seedling stage under 25% PEG-6000 treatment for 24 h. A total of 3927 proteins were accurately quantitated and 46, 36, 35, and 102 differentially abundant proteins (DAPs) were obtained in the S4, S4-1, T14, and T33 varieties, respectively. Four proteins were randomly selected for parallel reaction monitoring (PRM) assays, and the results verified the reliability of the mass spectrometry (MS) results. The response mechanism of the drought-sensitive sorghum leaves to drought was attributed to the upregulation of proteins involved in the tyrosine metabolism pathway with defense functions. Drought-resistant sorghum leaves respond to drought by promoting the TCA cycle, enhancing sphingolipid biosynthesis, interfering with triterpenoid metabolite synthesis, and influencing aminoacyl-tRNA biosynthesis. The 17 screened important candidate proteins related to drought stress were verified by quantitative real-time PCR (qRT-PCR), the results of which were consistent with the results of the proteomic analysis. This study lays the foundation for revealing the drought-resistance mechanism of sorghum at the protein level. These findings will help us cultivate and improve new drought-resistant sorghum varieties.

摘要

干旱是直接或间接抑制高粱生长、降低其生产力的主要限制因素。作为高粱的主要营养器官,叶片对干旱胁迫的响应机制在蛋白质组学水平上尚未阐明。本研究采用纳升级液相色谱-串联质谱(nano-LC-MS/MS)技术,比较了PEG-6000 处理 25%下,幼苗期抗旱性敏感(S4 和 S4-1)和抗旱性品种(T33 和 T14)叶片的蛋白质表达谱变化,处理 24 h。共准确定量了 3927 种蛋白质,在 S4、S4-1、T14 和 T33 品种中分别获得了 46、36、35 和 102 种差异丰度蛋白(DAP)。随机选择 4 种蛋白质进行平行反应监测(PRM)测定,结果验证了质谱(MS)结果的可靠性。抗旱性敏感高粱叶片对干旱的响应机制归因于防御功能相关的酪氨酸代谢途径上调。抗旱性高粱叶片通过促进三羧酸循环、增强鞘脂生物合成、干扰三萜类代谢物合成以及影响氨酰-tRNA 生物合成来应对干旱。通过定量实时 PCR(qRT-PCR)验证了与干旱胁迫相关的 17 个重要候选蛋白,结果与蛋白质组学分析结果一致。本研究为揭示高粱在蛋白质水平上的抗旱机制奠定了基础。这些发现将有助于我们培育和改良新的抗旱高粱品种。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0580/9656428/45dd82390708/ijms-23-13297-g001.jpg

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