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基于iTRAQ蛋白质组学筛选低温胁迫下影响藏红花成花的关键蛋白

Screening of Key Proteins Affecting Floral Initiation of Saffron Under Cold Stress Using iTRAQ-Based Proteomics.

作者信息

Chen Jing, Zhou Guifen, Dong Yan, Qian Xiaodong, Li Jing, Xu Xuting, Huang Huilian, Xu Limin, Li Liqin

机构信息

Huzhou Central Hospital, Affiliated Hospital of Huzhou Normal University, Huzhou, China.

Huzhou Hospital, Zhejiang University, Huzhou, China.

出版信息

Front Plant Sci. 2021 May 11;12:644934. doi: 10.3389/fpls.2021.644934. eCollection 2021.

DOI:10.3389/fpls.2021.644934
PMID:34046047
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC8144468/
Abstract

BACKGROUND

Saffron crocus () is an expensive and valuable species that presents preventive and curative effects. This study aimed to screen the key proteins affecting the floral initiation of saffron under cold stress and thus increasing yield by regulating the temperature.

RESULTS

Protein expression profiles in flowering and non-flowering saffron buds were established using isobaric tags for relative or absolute quantitation (iTRAQ). A total of 5,624 proteins were identified, and 201 differentially abundant protein species (DAPs) were further obtained between the flowering and non-flowering groups. The most important functions of the upregulated DAPs were "sucrose metabolic process," "lipid transport," "glutathione metabolic process," and "gene silencing by RNA." Downregulated DAPs were significantly enriched in "starch biosynthetic process" and several oxidative stress response pathways. Three new flower-related proteins, CsFLK, CseIF4a, and CsHUA1, were identified in this study. The following eight key genes were validated by real-time qPCR in flowering and non-flowering top buds from five different growth phases: floral induction- and floral organ development-related genes , , , and ; sucrose synthase activity-related genes and ; and starch synthase activity-related genes and . These findings demonstrate the important roles played by sucrose/starch biosynthesis pathways in floral development at the mRNA level. During normal floral organ development, the sucrose contents in the top buds of saffron increased, and the starch contents decreased. In contrast, non-flowering buds showed significantly decreased sucrose contents under cold stress and no significant changes in starch contents compared with those in the dormancy stage.

CONCLUSION

In this report, the protein profiles of saffron under cold stress and a normal environment were revealed for the first time by iTRAQ. A possible "reactive oxygen species-antioxidant system-starch/sugar interconversion flowering pathway" was established to explain the phenomenon that saffron does not bloom due to low temperature treatment.

摘要

背景

藏红花（）是一种昂贵且有价值的物种，具有预防和治疗作用。本研究旨在筛选在冷胁迫下影响藏红花花芽分化的关键蛋白质，从而通过调节温度提高产量。

结果

使用相对或绝对定量的等压标签（iTRAQ）建立了开花和未开花藏红花芽中的蛋白质表达谱。共鉴定出5624种蛋白质，开花组和未开花组之间进一步获得了201种差异丰富的蛋白质种类（DAPs）。上调的DAPs最重要的功能是“蔗糖代谢过程”、“脂质转运”、“谷胱甘肽代谢过程”和“RNA介导的基因沉默”。下调的DAPs在“淀粉生物合成过程”和几个氧化应激反应途径中显著富集。本研究鉴定出三种新的与花相关的蛋白质，CsFLK、CseIF4a和CsHUA1。通过实时qPCR对来自五个不同生长阶段的开花和未开花顶芽中的以下八个关键基因进行了验证：与花芽诱导和花器官发育相关的基因、、、和；与蔗糖合酶活性相关的基因和；以及与淀粉合酶活性相关的基因和。这些发现证明了蔗糖/淀粉生物合成途径在花发育的mRNA水平上所起的重要作用。在正常花器官发育过程中，藏红花顶芽中的蔗糖含量增加，淀粉含量降低。相比之下，在冷胁迫下，未开花芽的蔗糖含量显著降低，与休眠期相比，淀粉含量没有显著变化。

结论

在本报告中，首次通过iTRAQ揭示了冷胁迫和正常环境下藏红花的蛋白质谱。建立了一个可能的“活性氧-抗氧化系统-淀粉/糖相互转化开花途径”来解释藏红花因低温处理而不开花的现象。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/52c8/8144468/761ad657d6f1/fpls-12-644934-g006.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/52c8/8144468/a1c3e43fd8d3/fpls-12-644934-g001.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/52c8/8144468/6c3b6f4af462/fpls-12-644934-g002.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/52c8/8144468/fa16e901ec50/fpls-12-644934-g003.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/52c8/8144468/e08a5cf7fd32/fpls-12-644934-g004.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/52c8/8144468/83433d82512f/fpls-12-644934-g005.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/52c8/8144468/761ad657d6f1/fpls-12-644934-g006.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/52c8/8144468/a1c3e43fd8d3/fpls-12-644934-g001.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/52c8/8144468/6c3b6f4af462/fpls-12-644934-g002.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/52c8/8144468/fa16e901ec50/fpls-12-644934-g003.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/52c8/8144468/e08a5cf7fd32/fpls-12-644934-g004.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/52c8/8144468/83433d82512f/fpls-12-644934-g005.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/52c8/8144468/761ad657d6f1/fpls-12-644934-g006.jpg

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