Cloning and Functional Study of the LrMYB64 Gene in Lyciumruthenicum

doi:10.7525/j.issn.1673-5102.2026.02.004

Abstract

Abstract:

Lyciumruthenicum， a characteristic food-medicine plant in northwestern China， accumulates abundant anthocyanins in its fruits and possesses high pharmaceutical value. In order to identify the key regulators of anthocyanin biosynthesis and to clarify their coupled effects on lateral-bud development， the candidate gene LrMYB64 was cloned， an overexpression vector was constructed and transformed into L. ruthenicum， and the underlying molecular network was explored using transcriptome sequencing. The results showed that overexpression of LrMYB64 genein L. ruthenicum significantly suppressed the expression of key structural genes in the anthocyanin biosynthetic pathway-including LrCHS1B， LrCHS2， LrF3H， LrDFR， LrANS， and LrA3GT-leading to the reduction by 91.75% in anthocyanin level. Meanwhile， it markedly upregulated multiple hormone-signaling genes such as LrPR4， LrPP2C51， LrSAUR36， LrGH3.6， LrARF5， and LrABF4， increasing lateral bud outgrowth to approximately three times that of the wild type. Together， these findings indicated that LrMYB64 gene orchestrated a gene regulatory network through which it repressed anthocyanin biosynthesis while promoting hormone signal transduction， thereby coordinating plant secondary metabolism and hormonal responses.

Key words: Lyciumruthenicum, LrMYB64 gene, anthocyanin, lateral bud, transcriptome sequencing, functional analysis

CLC Number:

Q943.2

Yijiao LIU, Jiqing ZHAO, Jinhuan CHEN. Cloning and Functional Study of the LrMYB64 Gene in Lyciumruthenicum[J]. Bulletin of Botanical Research, 2026, 46(2): 231-245.

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引物名称 Primer name	引物序列（5′→3′） Primer sequence（5′→3′）	用途 Application
LrMYB64-F	ATGAGAAAACCTTGTTGTGATCA	全长克隆引物序列
LrMYB64-R	TTAAGGAAGTGAGTTGAGATCAGGC	全长克隆引物序列
35s-F	GACGCACAATCCCACTATCC	检测
35s-R	AATCATCGCAAGACCGGC	检测
LrMYB64-接头-F	GCGGGTCGACGGTACCATGAGAAAACCTTGTTGTG	重组引物
LrMYB64-接头-R	TAGACATATGGGTACCTTAAGGAAGTGAGTTGAGAT	重组引物

成分Component	体系System
黑果枸杞cDNA模板	1 μL
Forward primer	1 μL
Reverse primer	1 μL
PrimeSTAR MAX premix	10 μL
无菌蒸馏水	补足至20 μL

程序 Program	温度 Temperature	时间 Time	循环次数 Cycles
预变性	98 ℃	3 min	无
变性	98 ℃	30 s	30个循环
退火	58 ℃	30 s
延伸	72 ℃	2 min
延伸	72 ℃	10 min	无

引物名称 Primer name	引物序列（5′→3′） Primer sequence（5′→3′）	引物名称 Primer name	引物序列（5′→3′） Primer sequence（5′→3′）
Actin-7-F	CTGACGGTGAGGACATTCA	LrDFR-qPCR-F	AGGCTGCAATGGAAGAAGCT
Actin-7-R	GAGCATCATCTCCAGCAAAG	LrDFR-qPCR-R	AGCTAGGTGGGAACGTAGGT
LrFLS1-qPCR-F	TTGTCACTTGGGCTTGGGTT	LrCHI3-qPCR-F	CAGCAGTGGAAGGGCAAAAC
LrFLS1-qPCR-R	AAATCAGGCCTGGGACATGG	LrCHI3-qPCR-R	TGCACCCCATACTGTGAACC
LrF3H-qPCR-F	AAGGCATGTGAGGATTGGGG	LrPP2C51-qPCR-F	AGAAAGTCTCGCCACACAGG
LrF3H-qPCR-R	CAGTCTTGGACCACTTCCCC	LrPP2C51-qPCR-R	TTCCTCAACCAAGGTGTCGG
LrMYB64-qPCR-F	ACATGGTGAAGATTGCTGGAGA	LrGH3.6-like2-qPCR-F	TGTGCTTCGAGTTGCTGGAT
LrMYB64-qPCR-R	CTTCCAGCTATTAGGGACCATCT	LrGH3.6-like2-qPCR-R	GCACTGCATTGTTCACTGCA

温度 Temperature/℃	时间 Time/s	循环次数 Cycles
95	3	40个循环
98	5
60	34
95	15
60	60