Cloning of MeMYB106 in Cassava and Its Function in Anthocyanin Biosynthesis of Leaves

doi:10.13560/j.cnki.biotech.bull.1985.2025-0856

Abstract

Abstract:

Objective MYB transcription factors are primarily participate in the biosynthesis of flavonoids and anthocyanins. To explore MYB transcription factors in regulating anthocyanin biosynthesis in cassava leaves and verify its biological function would provide a theoretical basis for elucidating the molecular mechanisms of anthocyanin synthesis and leaves color improvement in cassava (Manihot esculenta Crantz). Method The coding sequence of MeMYB106 was cloned using cDNA from South China 9 (SC9) leaves as a template. Transient expression in tobacco was performed to determine its subcellular localization. RT-qPCR was applied to analyze the expression patterns of MeMYB106 in different tissues and differently colored leaves. Virus-induced gene silencing (VIGS) was employed to investigate the gene function in anthocyanin synthesis. Yeast one-hybrid assays and dual luciferase reporter assay were used to examine the regulatory relationship between MeMYB106 and anthocyanidin reductase (MeANR) gene. Result A MeMYB106 gene was cloned from SC9 leaves, with a coding region of 1 194 bp encoding 398 amino acids. The subcellular localization revealed that MeMYB106 was localized in the nucleus. Tissue-specific expression analysis showed that MeMYB106 was highly expressed in SC9 leaves, axillary buds, and stems, with significantly higher expression in green-leaves varieties compared with purple-leaves varieties. Silencing MeMYB106 in SC9 resulted in purple pigmentation in newly emerged leaves, with varying degrees of purple pigmentation depending on silencing efficiency. The anthocyanin content in the leaves of silenced plants was significantly higher than that in empty vector control. Further analysis demonstrated that MeMYB106 bound to the promoter region of MeANR to enhance its expression, thereby influencing anthocyanin biosynthesis in cassava leaves. Conclusion MeMYB106 transcription factor in cassava negatively regulate the biosynthesis of anthocyanins, and MeMYB106 positively regulate the expression of MeANR. Silencing MeMYB106 promotes anthocyanin accumulation in cassava leaves.

Key words: cassava, MeMYB106 transcription factor, anthocyanin, gene cloning, functional analysis

AN Fei-fei, LUO Xiu-qin, CAI Jie, XUE Jing-jing, ZHU Wen-li. Cloning of MeMYB106 in Cassava and Its Function in Anthocyanin Biosynthesis of Leaves[J]. Biotechnology Bulletin, 2026, 42(6): 13-21.

Figures/Tables 8

References 37

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引物名称 Primer name	引物序列 Primer sequence（5'-3'）	用途 Application
MeMYB106-F	ATGGGTCGATCGCCATGCTG	基因克隆 Gene clone
MeMYB106-R	TTAGAAGATGGGCGAATCTG	基因克隆 Gene clone
qPCR-MeMYB106-F	AATACTGTCGGTGGTCCTA	荧光定量PCR RT-qPCR
qPCR-MeMYB106-R	CTGAGAATGTTAGAGAAGATGTTG
qPCR-MeActin-F	TGATGAGTCTGGTCCATCCA
qPCR-MeActin-R	CCTCCTACGACCCAATCTCA
SubN-MeMYB106-F	agtggtctctgtccagtcctATGGGTCGATCGCCATGCTG	亚细胞定位 Subcellular localization
SubN-MeMYB106-R	ggtctcagcagaccacaagtTTAGAAGATGGGCGAATCTG
pGreen-SubN-F	CATGGTCCTGCTGGAGTTCGTG
pGreen-SubN-R	ACCGGCAACAGGATTCAATC
VIGS-MeMYB106-F	agtggtctctgtccagtcctGCCGCAGCGGTCTCTTGATG	基因沉默 Gene silencing
VIGS-MeMYB106-R	ggtctcagcagaccacaagtATATGATGATCTGAATTTCC
pCsCMV-F	TGGGCGCTAATTAGTTTACTGCA
pCsCMV-R	GGTCAAGACGGCTCAACTCTTCA
MeANR-promoter-F	agtggtctctgtccagtcctAAAAAGAATGAGAAAATT	启动子扩增 Promoter amplification
MeANR-promoter-R	ggtctcagcagaccacaagtTTGCAATTTTCCTTAAGG	启动子扩增 Promoter amplification
pNC-GADT7-F	TAATACGACTCACTATAGGG	酵母单杂交 Yeast one-hybrid
pNC-GADT7-F	AGATGGTGCACGATGCACAG
pNC-AbAi-F	CCTTCTGTTCGGAGATTACC
pNC-AbAi-R	TGCTACAAAGGACCTAATGT

引物名称 Primer name	引物序列 Primer sequence（5'-3'）	用途 Application
MeMYB106-F	ATGGGTCGATCGCCATGCTG	基因克隆 Gene clone
MeMYB106-R	TTAGAAGATGGGCGAATCTG	基因克隆 Gene clone
qPCR-MeMYB106-F	AATACTGTCGGTGGTCCTA	荧光定量PCR RT-qPCR
qPCR-MeMYB106-R	CTGAGAATGTTAGAGAAGATGTTG
qPCR-MeActin-F	TGATGAGTCTGGTCCATCCA
qPCR-MeActin-R	CCTCCTACGACCCAATCTCA
SubN-MeMYB106-F	agtggtctctgtccagtcctATGGGTCGATCGCCATGCTG	亚细胞定位 Subcellular localization
SubN-MeMYB106-R	ggtctcagcagaccacaagtTTAGAAGATGGGCGAATCTG
pGreen-SubN-F	CATGGTCCTGCTGGAGTTCGTG
pGreen-SubN-R	ACCGGCAACAGGATTCAATC
VIGS-MeMYB106-F	agtggtctctgtccagtcctGCCGCAGCGGTCTCTTGATG	基因沉默 Gene silencing
VIGS-MeMYB106-R	ggtctcagcagaccacaagtATATGATGATCTGAATTTCC
pCsCMV-F	TGGGCGCTAATTAGTTTACTGCA
pCsCMV-R	GGTCAAGACGGCTCAACTCTTCA
MeANR-promoter-F	agtggtctctgtccagtcctAAAAAGAATGAGAAAATT	启动子扩增 Promoter amplification
MeANR-promoter-R	ggtctcagcagaccacaagtTTGCAATTTTCCTTAAGG	启动子扩增 Promoter amplification
pNC-GADT7-F	TAATACGACTCACTATAGGG	酵母单杂交 Yeast one-hybrid
pNC-GADT7-F	AGATGGTGCACGATGCACAG
pNC-AbAi-F	CCTTCTGTTCGGAGATTACC
pNC-AbAi-R	TGCTACAAAGGACCTAATGT