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VvmiR160s/VvARFs 的相互作用及其在 GA 诱导葡萄单性结实过程中的时空表达/切割产物。

VvmiR160s/VvARFs interaction and their spatio-temporal expression/cleavage products during GA-induced grape parthenocarpy.

机构信息

College of Horticulture, Nanjing Agricultural University, Nanjing, 210095, China.

Department of Botany, Faculty of Sciences, Aswan University, Aswan, 81528, Egypt.

出版信息

BMC Plant Biol. 2019 Mar 21;19(1):111. doi: 10.1186/s12870-019-1719-9.

DOI:10.1186/s12870-019-1719-9
PMID:30898085
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC6429806/
Abstract

BACKGROUND

Grape (Vitis vinifera) is highly sensitive to gibberellin (GA), which effectively induce grape parthenocarpy. Studies showed that miR160s and their target AUXIN RESPONSIVE FACTOR (ARF) responding hormones are indispensable for various aspects of plant growth and development, but their functions in GA-induced grape parthenocarpy remain elusive.

RESULTS

In this study, the morphological changes during flower development in response to GA treatments were examined in the 'Rosario Bianco' cultivar. The precise sequences of VvmiR160a/b/c/d/e and their VvARF10/16/17 target genes were cloned, sequenced and characterized. The phylogenetic relationship and intron-exon structure of VvARFs and other ARF family members derived from different species were investigated. All VvmiR160s (except VvmiR160b) and VvARF10/16/17 had the common cis-elements responsive to GA, which support their function in GA-mediated grape parthenocarpy. The cleavage role of VvmiR160s-mediated VvARF10/16/17 was verified in grape flowers. Moreover, spatio-temporal expression analysis demonstrated that among VvmiR160 family, VvmiR160a/b/c highly expressed at late stage of flower/berry development, while VvARF10/16/17showed a reverse expression trend. Interestingly, GA exhibited a long-term effect through inducing the expression of VvmiR160a/b/c/e to increase their cleavage product accumulations from 5 to 9 days after treatment, but GA enhanced the expressions of VvARF10/16/17 only at short term. Pearson correlation analysis based on expression data revealed a negative correlation between VvmiR160a/b/c and VvARF10/16/17 in flowers not berries during GA-induced grape parthenocarpy.

CONCLUSIONS

This work demonstrated that the negative regulation of VvARF10/16/17 expression by VvmiR160a/b/c as key regulatory factors is critical for GA-mediated grape parthenocarpy, and provide significant implications for molecular breeding of high-quality seedless berry.

摘要

背景

葡萄（Vitis vinifera）对赤霉素（GA）极为敏感，GA 可有效诱导葡萄单性结实。研究表明，miR160s 及其靶基因 AUXIN RESPONSIVE FACTOR（ARF）响应激素在植物生长发育的各个方面都是不可或缺的，但它们在 GA 诱导葡萄单性结实中的功能仍不清楚。

结果

本研究以‘Rosario Bianco’品种为试材，探讨了 GA 处理对花发育过程中形态变化的影响。克隆、测序并分析了 VvmiR160a/b/c/d/e 的精确序列及其 VvARF10/16/17 靶基因。研究了来自不同物种的 VvARFs 和其他 ARF 家族成员的系统发育关系和内含子-外显子结构。所有 VvmiR160s（除 VvmiR160b 外）和 VvARF10/16/17 都具有响应 GA 的共同顺式元件，支持它们在 GA 介导的葡萄单性结实中的功能。葡萄花中验证了 VvmiR160s 介导的 VvARF10/16/17 的切割作用。此外，时空表达分析表明，在 VvmiR160 家族中，VvmiR160a/b/c 在花/浆果发育后期高表达，而 VvARF10/16/17 则呈现相反的表达趋势。有趣的是，GA 通过诱导 VvmiR160a/b/c/e 的表达来产生长期效应，从而增加其处理后 5-9 天的切割产物积累，但 GA 仅在短期增强 VvARF10/16/17 的表达。基于表达数据的 Pearson 相关分析表明，在 GA 诱导葡萄单性结实过程中，花而非浆果中 VvmiR160a/b/c 与 VvARF10/16/17 之间呈负相关。

结论

本研究表明，VvmiR160a/b/c 作为关键调节因子对 VvARF10/16/17 表达的负调控对于 GA 介导的葡萄单性结实至关重要，为高品质无核浆果的分子育种提供了重要启示。

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https://cdn.ncbi.nlm.nih.gov/pmc/blobs/e6b5/6429806/eda8a826f8c7/12870_2019_1719_Fig5_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/e6b5/6429806/9c1f884418ad/12870_2019_1719_Fig6_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/e6b5/6429806/643dc30ccfa1/12870_2019_1719_Fig7_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/e6b5/6429806/bc2735a28fc9/12870_2019_1719_Fig8_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/e6b5/6429806/69e5dfba1505/12870_2019_1719_Fig9_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/e6b5/6429806/e7b0f75d17d3/12870_2019_1719_Fig10_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/e6b5/6429806/2828dec183d4/12870_2019_1719_Fig11_HTML.jpg

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