Effect of R2R3-MYB Transcription Factor CitMYB21 on Flavonoids Biosynthesis in Citrus

doi:10.16420/j.issn.0513-353x.2021-1188

Acta Horticulturae Sinica ›› 2023, Vol. 50 ›› Issue (2): 250-264.doi: 10.16420/j.issn.0513-353x.2021-1188

• Research Papers • Previous Articles Next Articles

Effect of R2R3-MYB Transcription Factor CitMYB21 on Flavonoids Biosynthesis in Citrus

YE Zimao¹, SHEN Wanxia¹^,²^,^*(), LIU Mengyu¹, WANG Tong¹, ZHANG Xiaonan¹, YU Xin¹^,², LIU Xiaofeng¹^,², ZHAO Xiaochun¹^,²^,^*()

1. Citrus Research Institute，Southwest University/Chinese Academy of Agricultural Sciences，Chongqing 400712，China
2. National Citrus Engineering Research Center，Chongqing 400712，China

Received:2022-08-08 Revised:2022-10-19 Online:2023-02-25 Published:2023-03-06
Contact: *（E-mail：zhaoxiaochun@cric.cn，shenwanxia@cric.cn）

Abstract

Abstract:

Brazil Sour Orange（Citrus aurantium L.），Bendizao Mandarin（C. reticulata Blanco），Guihuadinanfeng mandarin（C. reticulata Blanco），Jincheng Beibei 447[C. sinensis（L.）Osbeck] and other 30 germplasms were used for mining the transcription factors related to flavonoid biosynthesis. Comparative transcriptomic analysis revealed that ERF，MYB and Dof transcription factor families are significantly involved in metabolism of flavonoids during fruit development. Correlation analysis of the expressions of CitCHS2，a key gene of flavonoids biosynthesis with the differentially expressed genes from transcriptomic analysis indicated that two R2R3-MYB family member CitMYB21（Ciclev10021699m）and CitMYB33（Ciclev10012152m）were negatively correlated to expression of CitCHS2. Phylogenic analysis revealed that these two genes belong to SG2 and SG1 subgroup of R2R3-MYBs family，respectively. The full length of CitMYB21 coding region is 804 bp and encodes 267 amino acids. The expressions of CitMYB21 were highly tissue and genotype specific，and significantly negatively correlated with the flavonoids contents. Silence of CitMYB21 resulted a significant increase of flavonoids in the leaf of transgenic plants，while transient overexpression of CitMYB21 in the peel of citrus fruit led to a significant decrease of flavonoids. Results of this study indicated that expression of CitMYB21 negatively affect the flavonoid biosynthesis in citrus，suggesting CitMYB21 may be an important transcription factor negatively regulating the biosynthesis of flavonoid.

Key words: citrus, flavonoids, R2R3-MYB transcription factor, negative regulation, tissue specific

CLC Number:

S666

YE Zimao, SHEN Wanxia, LIU Mengyu, WANG Tong, ZHANG Xiaonan, YU Xin, LIU Xiaofeng, ZHAO Xiaochun. Effect of R2R3-MYB Transcription Factor CitMYB21 on Flavonoids Biosynthesis in Citrus[J]. Acta Horticulturae Sinica, 2023, 50(2): 250-264.

Figures/Tables 14

References 45

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种类 Specific name	种质编号 Code	名称 Name	种类 Specific name	种质编号 Code	名称 Name
宽皮橘 Citrus reticulate Blanco	ZY004	桂花蒂南丰蜜橘 Guihuadinanfeng Mandarin	宽皮橘 C. reticulate Blanco	ZY198	半野生橘杂种No.3 Hybrid of Semi-wild No.3
	ZY013	槾橘 Manju		ZY208	无核早橘 Seedless Zaoju
	ZY014	野橘 2 Yeju 2		ZY215	丙中洛橘 Bingzhongluoju
	ZY026	酸橘 Suanju		ZY222	红橘 Hongju
	ZY035	鹅蛋香柑 E Dan Xianggan		ZY007	津优 Jinyou
	ZY036	汉源黄果柑 Hanyuan Huangguogan		ZY008	扁柑 Biangan
	ZY037	德昌金钱橘 Dechang Jinqianju		ZY011	久贺早生温州 Kuga Wase
	ZY049	冰糖香柑 Bingtang Xianggan		ZY012	北口温州 Kitaguchi Wase
	ZY084	青红橘 Qinghongju	温州蜜柑 C. unshiu Macf.	ZY067	久能温州 Kunou Wase
	ZY085	红橙 Hongcheng		ZY070	繁田温州 Shigeta Wase
	ZY090	元阳酸橘 Yuanyang Suanju		ZY124	大分1号温州 Dafen No.1 Wase
	ZY108	沅江南橘 Yuanjiang Nanju		ZY145	湘慈43号 Xiangci No.43
	ZY119	马塔里橘 Matari Tangerine		ZY168	谷口系日南1号温州
	ZY151	新生系3号椪柑 No.3 Ponkan			Taniguchi Line Nichinan No.1 Wase
	ZY174	春见 Harumi		ZY231	桥本温州 Hashimoto Wase
				ZY002	平阳橘 Duong Mandarin

种类 Specific name	种质编号 Code	名称 Name	种类 Specific name	种质编号 Code	名称 Name
宽皮橘 Citrus reticulate Blanco	ZY004	桂花蒂南丰蜜橘 Guihuadinanfeng Mandarin	宽皮橘 C. reticulate Blanco	ZY198	半野生橘杂种No.3 Hybrid of Semi-wild No.3
	ZY013	槾橘 Manju		ZY208	无核早橘 Seedless Zaoju
	ZY014	野橘 2 Yeju 2		ZY215	丙中洛橘 Bingzhongluoju
	ZY026	酸橘 Suanju		ZY222	红橘 Hongju
	ZY035	鹅蛋香柑 E Dan Xianggan		ZY007	津优 Jinyou
	ZY036	汉源黄果柑 Hanyuan Huangguogan		ZY008	扁柑 Biangan
	ZY037	德昌金钱橘 Dechang Jinqianju		ZY011	久贺早生温州 Kuga Wase
	ZY049	冰糖香柑 Bingtang Xianggan		ZY012	北口温州 Kitaguchi Wase
	ZY084	青红橘 Qinghongju	温州蜜柑 C. unshiu Macf.	ZY067	久能温州 Kunou Wase
	ZY085	红橙 Hongcheng		ZY070	繁田温州 Shigeta Wase
	ZY090	元阳酸橘 Yuanyang Suanju		ZY124	大分1号温州 Dafen No.1 Wase
	ZY108	沅江南橘 Yuanjiang Nanju		ZY145	湘慈43号 Xiangci No.43
	ZY119	马塔里橘 Matari Tangerine		ZY168	谷口系日南1号温州
	ZY151	新生系3号椪柑 No.3 Ponkan			Taniguchi Line Nichinan No.1 Wase
	ZY174	春见 Harumi		ZY231	桥本温州 Hashimoto Wase
				ZY002	平阳橘 Duong Mandarin

引物名称 Primers name	序列（5′-3′） Sequence	酶切位点 Restriction site
OE-CitMYB21F	CCATTTAAATATGGTGAGAGCTCCATGCTG	SwaⅠ
OE-CitMYB21R	CGGGATCCCGACTTCTCATCAAAAAGGAAGGA	BamHⅠ
VIGS-CitMYB21F	TCCCCCGGGACAACAAGAGCCCGTGATTA	SmaⅠ
VIGS-CitMYB21R	CGGGATCCAACTATAGACAGTGGGTCGGC	BamHⅠ
q-CitMYB21F	CATGCTGCGAGAAGATGGGA
q-CitMYB21R	CTGCTTGTTTGGGAAGTGCC
Actin-F	CATCCCTCAGCACCTTCC
Actin-R	CCAACCTTAGCACTTCTCC
pTRV1-F	TTGGGTTGCTACTGATTCGACT
pTRV1-R	CTGTAAGGACCATCATACTTCGC
pTRV2_GFP-F	CTGCCCGACAACCACTACCT
pTRV2_GFP-R	CTTGTACAGCTCGTCCATGCC

引物名称 Primers name	序列（5′-3′） Sequence	酶切位点 Restriction site
OE-CitMYB21F	CCATTTAAATATGGTGAGAGCTCCATGCTG	SwaⅠ
OE-CitMYB21R	CGGGATCCCGACTTCTCATCAAAAAGGAAGGA	BamHⅠ
VIGS-CitMYB21F	TCCCCCGGGACAACAAGAGCCCGTGATTA	SmaⅠ
VIGS-CitMYB21R	CGGGATCCAACTATAGACAGTGGGTCGGC	BamHⅠ
q-CitMYB21F	CATGCTGCGAGAAGATGGGA
q-CitMYB21R	CTGCTTGTTTGGGAAGTGCC
Actin-F	CATCCCTCAGCACCTTCC
Actin-R	CCAACCTTAGCACTTCTCC
pTRV1-F	TTGGGTTGCTACTGATTCGACT
pTRV1-R	CTGTAAGGACCATCATACTTCGC
pTRV2_GFP-F	CTGCCCGACAACCACTACCT
pTRV2_GFP-R	CTTGTACAGCTCGTCCATGCC

转化方式 Transformation method	r 相关性Correlation	P 显著性Significance
病毒诱导的基因沉默Virus induced gene slicing	-0.8	< 0.01
瞬时表达Transient expression	-0.8	> 0.05

Effect of R2R3-MYB Transcription Factor CitMYB21 on Flavonoids Biosynthesis in Citrus

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基因 Gene	CitPAL	CitC4H	Cit4CL1	Cit4CL2	CitCHS2	CitCHI	条件 Condition
CitMYB21	-0.7^**	0.3	-0.5^*	0.3	-0.4	-0.5	叶片发育早期Leaf early developmental stage
CitMYB21	0.1	-0.4	0.5^*	-0.5	0.6^*	0.4	叶片发育后期Leaf late developmental stage
CitMYB21	0.1	0.9^**	-0.6	0.7	-0.6	0.8^**	组织部位Tissue
CitMYB21	0.3	0.6^*	-0.4	-0.5	-0.7^**	-0.6^**	基因沉默Gene silencing
CitMYB21	0.2	0.8	-0.3^**	0.7	-0.9^**	-0.9^**	瞬时表达Transient expression

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