Csn-miR156d-CsSPL1 regulates flowering and anthocyanin metabolism.

IF 3.5 2区 农林科学 Q1 FORESTRY
Qingqing Lin, Hui Li, Hu He, Pu Wang, Mingle Wang, Hua Zhao, Yu Wang, Dejiang Ni, Yanni Fang, Fei Guo
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Abstract

MiR156 play important roles in regulation of plant growth and development, secondary metabolite synthesis, and other biological processes by targeting the SQUAMOSA promoter binding protein-like (SPL) family. Our previous sequencing data analysis suggested that Csn-miR156d may regulate flowering and anthocyanin accumulation by cleavage and degradation of the expression of the SPL in tea plant, but it remains to be elucidated. In this study, 5'RLM-RACE experiment, tobacco transient transformation, qRT-PCR, and antisense oligonucleotide (asODN) were used to verify that CsSPL1 is the target gene of Csn-miR156d. Stable transformation of Arabidopsis revealed that Csn-miR156d could delay flowering by negatively regulating the transcript levels of FT, AP1, FUL, and SOC1, while overexpression of CsSPL1 showed an opposite effect. Additionally, overexpression of Csn-miR156d in Arabidopsis could enhance the transcription of the anthocyanin biosynthesis-related structural genes DFR, ANS, F3H, UGT78D2, and LDOX, as well as regulatory genes PAP1, MYB113, GL3, MYB11, and MYB12, leading to anthocyanin accumulation. Moreover, asODN experiment revealed that Csn-miR156d could increase the anthocyanin content in tea plant. These results suggest that Csn-miR156d regulates flowering and anthocyanin accumulation in tea plant by suppressing the expression of CsSPL1. Our study provides new insights into the development and anthocyanin accumulation in tea plant and lays a theoretical foundation for further research on the molecular mechanism of miRNAs in regulating tea plant growth and secondary metabolism.

Csn-miR156d-CsSPL1调控开花和花青素代谢。
MiR156通过靶向SQUAMOSA类启动子结合蛋白(SPL)家族,在调控植物生长发育、次生代谢产物合成等生物过程中发挥重要作用。我们之前的测序数据分析表明,Csn-miR156d可能通过裂解和降解茶树中SPL的表达来调控开花和花青素的积累,但这一观点还有待进一步阐明。本研究采用5'RLM-RACE实验、烟草瞬时转化、qRT-PCR和反义寡核苷酸(asODN)等方法验证了CsSPL1是Csn-miR156d的靶基因。拟南芥的稳定转化结果表明,Csn-miR156d可通过负调控FT、AP1、FUL和SOC1的转录水平来延迟开花,而过表达CsSPL1则显示出相反的效果。此外,在拟南芥中过表达 Csn-miR156d 可增强花青素生物合成相关结构基因 DFR、ANS、F3H、UGT78D2 和 LDOX 以及调控基因 PAP1、MYB113、GL3、MYB11 和 MYB12 的转录,从而导致花青素积累。此外,asODN 实验表明,Csn-miR156d 能增加茶树中的花青素含量。这些结果表明,Csn-miR156d通过抑制CsSPL1的表达调控茶树开花和花青素积累。我们的研究为茶树的生长发育和花青素积累提供了新的见解,为进一步研究miRNA调控茶树生长和次生代谢的分子机制奠定了理论基础。
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来源期刊
Tree physiology
Tree physiology 农林科学-林学
CiteScore
7.10
自引率
7.50%
发文量
133
审稿时长
1 months
期刊介绍: Tree Physiology promotes research in a framework of hierarchically organized systems, measuring insight by the ability to link adjacent layers: thus, investigated tree physiology phenomenon should seek mechanistic explanation in finer-scale phenomena as well as seek significance in larger scale phenomena (Passioura 1979). A phenomenon not linked downscale is merely descriptive; an observation not linked upscale, might be trivial. Physiologists often refer qualitatively to processes at finer or coarser scale than the scale of their observation, and studies formally directed at three, or even two adjacent scales are rare. To emphasize the importance of relating mechanisms to coarser scale function, Tree Physiology will highlight papers doing so particularly well as feature papers.
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