揭示microRNAs介导的葡萄果实采后衰老调控网络

doi:10.1016/j.jia.2024.12.039

摘要/Abstract

摘要：

鲜食葡萄的采后衰老阶段包含一系列生物学过程。MicroRNAs（miRNAs）在转录后水平调控下游基因的表达；然而，miRNAs是否参与葡萄采后衰老过程仍不明确。本研究采用小RNA高通量测序技术，鉴定了‘红地球’（Vitis vinifera）葡萄在采收后4°C条件下贮藏0，30和60天后（分别记为RG0、RG30、RG60）衰老相关miRNAs差异变化。结果共鉴定出42个已知的miRNAs和219个新的候选miRNAs。在果实衰老过程中，PC-3p-3343_1921、miR2950、miR395k、miR2111、miR159c、miR169q、PC-5p-1112_4500和miR167b的表达水平发生显著变化（p<0.05）。降解组测序共鉴定出218个靶基因，涉及细胞壁组织、三羧酸循环、病害防御、碳代谢、激素信号传导、花青素代谢途径和能量调控等过程，其中ARF6, GRF3, TCP2, CP1, MYBA2和WRKY72与果实衰老密切相关。此外，本研究还通过双荧光素酶报告基因实验和葡萄果实瞬时转化实验，验证了PC-5p-1112_4500靶向剪切的3个功能未知的靶基因（VIT_00s2146g00010、VIT_02s0012g01750和VIT_03s0038g00160），并证实了它们具有调控果实衰老的功能。这些结果深化了人们对miRNAs在调控葡萄果实衰老和延长园艺产品货架期方面作用的认识。基于这些发现，本研究提出了一种新的理论策略，即通过调控关键miRNAs（例如PC-5p-1112_4500）的表达来延缓园艺产品的采后衰老，从而延长其货架期。

Abstract:

The postharvest senescence phase of table grapes comprises a series of biological processes. MicroRNAs (miRNAs) regulate downstream genes at the post-transcriptional level; however, whether miRNAs are involved in postharvest grape senescence remains unclear. We used small RNA sequencing to identify postharvest-related miRNAs in ‘Red Globe’ (Vitis vinifera) grapes harvested after 0, 30, and 60 d at 4 °C (RG0, RG30, RG60). In total, 42 known and 219 novel miRNA candidates were obtained. During fruit senescence, the expression of PC-3p-3343_1921, miR2950, miR395k, miR2111, miR159c, miR169q, PC-5p-1112_4500, and miR167b changed significantly (p<0.05). Degradation sequencing identified 218 targets associated with cell wall organization, TCA cycling, pathogen defense, carbon metabolism, hormone signaling, the anthocyanin metabolism pathway, and energy regulation, of which ARF6, GRF3, TCP2, CP1, MYBA2, and WRKY72 were closely related to fruit senescence. We also verified VIT_00s2146g00010, VIT_02s0012g01750, and VIT_03s0038g00160 with unknown functions cleaved by senescence-related PC-5p-1112_4500 via the dual luciferase assay and transient transformation of grape berries and showed that they regulate berry senescence. These results deepen the understanding of the contribution of miRNAs in regulating grape berry senescence and prolonging the shelf life of horticultural products. Based on these results, we propose a new theoretical strategy to delay the postharvest senescence of horticultural products by regulating the expression of key miRNAs (e.g., PC-5p-1112_4500), thereby extending their shelf life.

Key words: ‘Red Globe’ berries , small RNA library , cleaved transcripts , fruit ripening , senescence regulation , post-transcriptional regulators

. 揭示microRNAs介导的葡萄果实采后衰老调控网络[J]. Journal of Integrative Agriculture, DOI: 10.1016/j.jia.2024.12.039.

Mingxin Feng, Ying Hu, Xin Yang, Jingwen Li, Haochen Wang, Yujia Liu, Haijun Ma, Kai Li, Jiayin Shang, Yulin Fang, Jiangfei Meng. Uncovering the miRNA-mediated regulatory network involved in postharvest senescence of grape berries[J]. Journal of Integrative Agriculture, DOI: 10.1016/j.jia.2024.12.039.

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