A comprehensive investigation of starch degradation process and identification of a transcriptional activator MabHLH6 during banana fruit ripening.
A comprehensive investigation of starch degradation process and identification of a transcriptional activator MabHLH6 during banana fruit ripening.
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香蕉果实成熟过程中淀粉降解过程的全面研究和转录激活子 MabHLH6 的鉴定。
DOI：
10.1111/pbi.12756
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发表时间：
2018-01
影响因子：
13.8
通讯作者：
Lu WJ
中科院分区：
文献类型：
--
作者：
Xiao YY;Kuang JF;Qi XN;Ye YJ;Wu ZX;Chen JY;Lu WJ
虽然淀粉降解已经在拟南芥叶片和谷类种子等模型系统中得到了很好的研究，但在淀粉类水果，特别是香蕉成熟过程中的这一过程在很大程度上是未知的。在这项研究中，从香蕉果实中鉴定和鉴定了38个编码淀粉降解相关蛋白的基因。表达分析显示，27个候选基因在香蕉果实成熟过程中被显著诱导，伴随着淀粉向糖的转化。此外，基于iTRAQ的蛋白质组学实验确定了18种与淀粉颗粒表面结合的淀粉降解相关酶，其中10种在成熟过程中显著上调。更重要的是，基于使用MaGWD 1启动子作为诱饵的酵母单杂交筛选，鉴定了一种新的bHLH转录因子MabHLH 6。在果实成熟过程中，MabHLH 6的转录和蛋白质水平也有所增加。电泳迁移率变动分析、染色质免疫沉淀和瞬时表达实验证实，MabHLH 6激活11个淀粉降解相关基因的启动子，包括Ma GWD 1、Ma LSF 2、Ma BAM 1、Ma BAM 2、Ma BAM 8、Ma BAM 10、Ma AMY 3、Ma AMY 3 C、Ma伊萨2、Ma伊萨3和MapGlcT 2 - 2通过识别它们存在于启动子中的E-盒（CANNTG）基序来表达。总的来说，这些研究结果表明，香蕉果实成熟过程中的淀粉降解可能归因于转录和翻译水平上与淀粉分解相关的许多酶的复杂作用，并且MabHLH 6可能通过直接激活一系列淀粉降解相关基因而作为该过程的正调节因子。
Although starch degradation has been well studied in model systems such as Arabidopsis leaves and cereal seeds, this process in starchy fruits during ripening, especially in bananas, is largely unknown. In this study, 38 genes encoding starch degradation‐related proteins were identified and characterized from banana fruit. Expression analysis revealed that 27 candidate genes were significantly induced during banana fruit ripening, with concomitant conversion of starch‐to‐sugars. Furthermore, iTRAQ‐based proteomics experiments identified 18 starch degradation‐associated enzymes bound to the surface of starch granules, of which 10 were markedly up‐regulated during ripening. More importantly, a novel bHLH transcription factor, MabHLH6, was identified based on a yeast one‐hybrid screening using MaGWD1 promoter as a bait. Transcript and protein levels of MabHLH6 were also increased during fruit ripening. Electrophoretic mobility shift assays, chromatin immunoprecipitation and transient expression experiments confirmed that MabHLH6 activates the promoters of 11 starch degradation‐related genes, including Ma GWD 1, Ma LSF 2, Ma BAM 1, Ma BAM 2, Ma BAM 8, Ma BAM 10, Ma AMY 3, Ma AMY3 C, Ma ISA 2, Ma ISA 3 and MapGlcT2‐2 by recognizing their E‐box (CANNTG) motifs present in the promoters. Collectively, these findings suggest that starch degradation during banana fruit ripening may be attributed to the complex actions of numerous enzymes related to starch breakdown at transcriptional and translational levels, and that MabHLH6 may act as a positive regulator of this process via direct activation of a series of starch degradation‐related genes.
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