Characterization of Vv-miR156: Vv-SPL pairs involved in the modulation of grape berry development and ripening
Characterization of Vv-miR156: Vv-SPL pairs involved in the modulation of grape berry development and ripening
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DOI：
10.1007/s00438-018-1462-1
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发表时间：
2018-12-01
影响因子：
3.1
通讯作者：
Fang, Jinggui
中科院分区：
文献类型：
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作者：
Cui, Mengjie;Wang, Chen;Fang, Jinggui
SPL是一个植物特异性转录因子家族。许多研究人员报道，miR156s靶向的SPL成员可能在植物生长发育的调节中发挥重要作用。虽然关于葡萄也有类似的报道，但到目前为止，人们对葡萄果实的发育和成熟知之甚少。为了进一步了解葡萄miR156s （Vv-miR156s）如何通过介导靶基因Vv-SPL调控葡萄果实的上述过程，我们鉴定了巨玫瑰葡萄果实中Vv-miR156s的精确序列，预测了它们的潜在靶标，并揭示了各种Vv-miR156: Vv-SPL对的匹配程度存在一定差异，表明它们的相互作用存在差异。随后，我们还在Vv-MIR156s和Vv-SPLs的启动子中发现了类似的基序，如ABRE、CGTCA和ERE，它们对浆果的发育和成熟更具特异性。随着浆果的发育和成熟，Vv-miR156a、b/c/d、e和f/g/i的表达量总体呈上升趋势，而它们对应阶段的靶基因表达量呈相反趋势。此外，外源ABA和NAA对葡萄果实成熟期前Vv-miR156s的表达均有一定的促进或抑制作用。利用本研究开发的PPM-RACE和改良的rlc - race结合qRT-PCR验证了Vv-miR156a、b/c/d、e、f/g/i及其靶基因（Vv-SPL2、9、10、16）在葡萄果实发育和成熟过程中的裂解产物、位点和频率，表明Vv-miR156s可以通过介导Vv-SPL2、9、10、16的表达参与葡萄果实发育和成熟过程的调控。本研究结果为进一步认识其在葡萄果实中的功能奠定了重要基础，丰富了对mirna介导的葡萄果实发育成熟调控机制的认识。
SPL is a plant-specific transcription factor family. Many researchers reported that SPL members targeted by miR156s could play crucial roles in the modulation of plant growth and development. Although there are similar reports on grapes, till now little is known about grape berry development and ripening. To gain more insight into how grape miR156s (Vv-miR156s) modulated the above given processes of grape berries by mediating their target gene Vv-SPLs, here we identified the precise sequences of Vv-miR156s in Giant Rose' grape berries, predicted their potential targets, and revealed that the matching degree of various Vv-miR156: Vv-SPL pairs exhibited some discrepancy, implying the divergence of their interaction. Subsequently, we also discovered similar motifs such as ABRE, CGTCA and ERE, which are more specific to berry development and ripening, within the promoters of both Vv-MIR156s and Vv-SPLs. With berry development and ripening, meanwhile, Vv-miR156a, b/c/d, e and f/g/i exhibited an overall increasing expression trend, while their targets showed opposite trends at the corresponding stages. Additionally, exogenous ABA and NAA application promoted or curbed the expression of Vv-miR156s to some extent, before grape berry ripening stage. The cleavage products, sites and frequencies of Vv-miR156a, b/c/d, e, f/g/i and their respective targets (Vv-SPL2, 9, 10, 16) during grape berry development and ripening process were validated by our developed PPM-RACE and modified RLM-RACE together with qRT-PCR, which demonstrated that Vv-miR156s can be involved in the modulation of grape berry development and ripening process by mediating the expression of Vv-SPL2, 9, 10, 16. Our findings lay an important foundation for further recognizing their functions in grape berries, and enrich the knowledge of the regulatory mechanism of miRNA-mediated grape berry development and ripening.