光信号转导因子HY5调控拟南芥分枝的功能研究

doi:10.19586/j.2095-2341.2021.0102

生物技术进展 ›› 2022, Vol. 12 ›› Issue (3): 379-386.DOI: 10.19586/j.2095-2341.2021.0102

光信号转导因子HY5调控拟南芥分枝的功能研究

周琴(), 谢钰容()

中国农业科学院生物技术研究所，北京 100081

收稿日期:2021-06-04 接受日期:2021-11-08 出版日期:2022-05-25 发布日期:2022-05-26
通讯作者: 谢钰容
作者简介:周琴 E-mail：2777314088@qq.com；
基金资助:
北京市自然科学基金项目(5222030);国家自然科学基金项目(31770210)

Functional Analysis of ELONGATED HYPOCOTYL5 Regulating Branching in Arabidopsis thaliana

Qin ZHOU(), Yurong XIE()

Biotechnology Research Institute，Chinese Academy of Agricultural Sciences，Beijing 100081，China

Received:2021-06-04 Accepted:2021-11-08 Online:2022-05-25 Published:2022-05-26
Contact: Yurong XIE

摘要/Abstract

摘要：

光是影响植物分枝的重要外在环境因素，但光信号因子HY5（ELONGATED HYPOCOTYL5）是否调控植物分枝目前尚不清楚。创制了HY5转基因超表达植株，并获得商业化T-DNA插入突变体纯合植株。通过比较野生型（WT）、超表达植株（HY5-OE）、突变体（hy5-215）的分枝数目发现，与野生型相比，超表达植株分枝数目显著增加，而突变体分枝数目则显著减少。进一步比较这些遗传材料的拟南芥植株分枝的负调控关键因子BRC1（BRANCHED1）转录本水平差异，发现与野生型相比，超表达植株中BRC1转录本显著下调、突变体中显著上调。研究结果表明，HY5通过抑制拟南芥分枝关键负调控因子BRC1的转录水平，进而促进拟南芥的分枝。研究结果为阐明HY5调控分枝的生物学功能提供了一定的理论依据。

关键词: 拟南芥, 光信号转导因子HY5, 分枝

Abstract:

Light is one of the most important external environmental factors that affects plant branching， however， whether ELONGATED HYPOCOTYL5 （HY5） regulating plant branching is still unknown. In this study， HY5 transgenic overexpression plants were created and commercial T-DNA insertion mutant homozygous plants were obtained. By comparing the number of branches of wild type （WT）， overexpression plant （HY5-OE） and mutant （hy5?215）， the results showed that the overexpression plants had much more rosette branches， while the mutants had less branches compared with wild type plants. The transcriptional levels of BRC1 （BRANCHED1）， a key negative regulatory factor of Arabidopsis thaliana branches were further compared. The results also showed that BRC1 transcription was significantly down-regulated in overexpressed plants and up-regulated in mutants. The results suggested that HY5 promotes Arabidopsis thaliana branching by inhibiting the transcription of BRC1. These results of this research provided a theoretical basis for elucidating the biological functions of HY5 regulatory branches.

Key words: Arabidopsis thaliana, ELONGATED HYPOCOTYL5, branching

中图分类号:

Q943

周琴, 谢钰容. 光信号转导因子HY5调控拟南芥分枝的功能研究[J]. 生物技术进展, 2022, 12(3): 379-386.

Qin ZHOU, Yurong XIE. Functional Analysis of ELONGATED HYPOCOTYL5 Regulating Branching in Arabidopsis thaliana[J]. Current Biotechnology, 2022, 12(3): 379-386.

图/表 8

表1 实验所用引物

Table1 Primers used in the experiment

引物名称	引物序列（5'→3'）	引物用途
HY5‑GFP‑F	5'‑GTTTGGTGTTACTTCTGCAGATGCAGGAACAAGCGACTAGCT‑3'	扩增HY5编码区序列
HY5‑GFP‑R	5'‑TCGAGTCGACCCGGGAATTCGAAGGCTTGCATCAGCATTAG‑3'	扩增HY5编码区序列
HY5‑qF	5'‑CGATGAGGAGATACGGCGAG‑3'	RT⁃qPCR检测HY5基因表达量
HY5‑qR	5'‑AACCTCTTCAGCCGCTTGTT‑3'	RT⁃qPCR检测HY5基因表达量
PP2A‑F	5'‑TATCGGATGACGATTCTTCGTGCAG‑3'	RT⁃qPCR检测PP2A基因表达量
PP2A‑R	5'‑GCTTGGTCGACTATCGGAATGAGAG‑3'	RT⁃qPCR检测PP2A基因表达量
hy5‑215‑F	5'‑ATCAAGCAGCGAGAGGTCAT‑3'	hy5⁃215突变体鉴定
hy5‑215‑R	5'‑TTCAGCCGCTTGTTCTCTTT	hy5⁃215突变体鉴定
hy5‑salk‑LP	5'‑ATTCCTTCCCAAAATGTCTCG‑3'	hy5⁃10突变体鉴定
hy5‑salk‑RP	5'‑ATGCGAGTGAATGACCATTTC‑3'
LB	5'‑ATTTTGCCGATTTCGGAAC‑3'
BRC1‑qF	5'‑CCATTGCTCCCTTTTAGCCC‑3'	RT⁃qPCR检测BRC1基因表达量
BRC1‑qR	5'‑CTGGGTGGAAAGAGGTGAGG‑3'	RT⁃qPCR检测BRC1基因表达量

图1 HY5?GFP过表达载体

Fig.1 Vector of HY5?GFP

图2 转基因阳性植株的筛选

Fig.2 Screening of transgenic positive plants

图3 HY5?GFP过表达植株激发光检测

Fig.3 Fluorescence detection of HY5?GFP overexpression plants

图4 转基因株系HY5相对表达量注：**表示与野生型相比差异有统计学意义（P<0.01）。

Fig.4 Relative expression of HY5 in transgenic lines

图5 hy5突变体鉴定A：hy5?215突变体序列比对；B：hy5?10突变体鉴定

Fig.5 Identification of hy5 mutant

图6 拟南芥各基因型表型分析A：分枝表型图；B：各基因型株系分枝数目统计。**表示与野生型相比差异有统计学意义（P<0.01）。

Fig.6 Analysing of branching phenotype of Arabidopsis thaliana

图7 BRC1表达水平检测注：**表示与野生型相比差异有统计学意义（P<0.01）。

Fig.7 Detection of expression of BRC1

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光信号转导因子HY5调控拟南芥分枝的功能研究

Functional Analysis of ELONGATED HYPOCOTYL5 Regulating Branching in Arabidopsis thaliana

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