PavMYB10.1 Promotes Anthocyanin Accumulation by Positively Regulating PavRiant in Sweet Cherry

doi:10.16420/j.issn.0513-353x.2021-0732

Acta Horticulturae Sinica ›› 2022, Vol. 49 ›› Issue (5): 1023-1030.doi: 10.16420/j.issn.0513-353x.2021-0732

• Research Papers • Previous Articles Next Articles

PavMYB10.1 Promotes Anthocyanin Accumulation by Positively Regulating PavRiant in Sweet Cherry

LI Lixian¹, WANG Shuo¹, CHEN Ying¹, WU Yingtao¹, WANG Yaqian¹, FANG Yue¹, CHEN Xuesen¹, TIAN Changping², FENG Shouqian¹^,^*()

¹State Key Laboratory of Crop Biology,College of Horticulture Science and Engineering,Shandong Agricultural University,Tai’an,Shandong 271018,China
²Yantai Academy of Agricultural Sciences,Yantai,Shandong 265599,China

Received:2022-01-04 Revised:2022-03-28 Online:2022-05-25 Published:2022-05-25
Contact: FENG Shouqian E-mail:shouqianlove@sdau.edu.cn

Abstract

Abstract:

To understand how GST regulates anthocyanin accumulation in sweet cherry,a phi-type GST gene PavRiant（XM_021968160）was cloned from the sweet cherry cultivar‘Tieton’. The effects of PavRiant on anthocyanin accumulation in sweet cherry were investigated using phylogenetic tree analysis,real-time reverse transcription-polymerase chain reaction,transient silencing assay,yeast one-hybrid（Y1H）,electrophoretic mobility shift assay（EMSA）,and luciferase reporter assay. Phylogenetic tree analysis showed that PavRiant has the highest similarity to the PpRiant protein sequence of peach. During sweet cherry fruit development,the changing trends of PavRiant expressions were consistent with anthocyanin contents and PavMYB10.1 expressions. Further,the transient silencing assay showed that PavRiant plays an essential role in regulating anthocyanin accumulation. Y1H and EMSA experiments showed that PavMYB10.1 binds to the MRE sequence of the PavRiant promoter. Finally,the luciferase assay showed that PavMYB10.1 positively regulates the activity of the PavRiant promoter. Therefore,PavMYB10.1 can promote anthocyanin accumulation by positively regulating PavRiant expression in sweet cherry.

Key words: sweet cherry, anthocyanin, PavMYB10.1, PavRiant, GST

CLC Number:

S662.5

LI Lixian, WANG Shuo, CHEN Ying, WU Yingtao, WANG Yaqian, FANG Yue, CHEN Xuesen, TIAN Changping, FENG Shouqian. PavMYB10.1 Promotes Anthocyanin Accumulation by Positively Regulating PavRiant in Sweet Cherry[J]. Acta Horticulturae Sinica, 2022, 49(5): 1023-1030.

Figures/Tables 6

Table 1 The list of primer sequences

用途 Purpose	引物名称 Primer name	序列（5-3′） Sequence
瞬时沉默试验 Transient silencing assay	PavRiant-（TRV2）-F	GAATTCTCCCAACCTATTGGGAACAACC
瞬时沉默试验 Transient silencing assay	PavRiant-（TRV2）-R	GGTACCGTAGTCACTAGCAAGGCTCATTAG
荧光实时定量 PCR qRT-PCR	PavRiant-F	GCTATCTGGCTGGAGAAG
	PavRiant-R	GCTATCTGGCTGGAGAAG
	PavActin-F	CCAGGGCTGTGTTTCCTTCTATT
	PavActin-R	ATGATCTGCGTCATCTTTTCT
电泳迁移率试验 EMSA	PavMYB10.1-（PGEX4T）-F	GGATCCATGGCCAACAAGAAGATGGAGG
电泳迁移率试验 EMSA	PavMYB10.1-（PGEX4T）-R	GAATTCGTCCTTCTGAACATTGGTACACTGTC
荧光素酶报告基因试验 LUC	PavMYB10.1-（62-SK）-F	TGGATCCATGGCCAACAAGAAGATGGAGG
	PavMYB10.1-（62-SK）-R	AAGCTTGTCCTTCTGAACATTGGTACACTGTC
	PavRiant-（0800-LUC）-F	GTCGACTCGAATTGGAGTTTGTTCGAAA
	PavRiant-（0800-LUC）-R	GGATCCTTCTAGACATAAAAGTAAACATTAAATTATATAAACC
酵母单杂交试验 Y1H	PavRiant-（PHIS2）-F	GAATTCTCGAATTGGAGTTTGTTCGAAA
	PavRiant-（PHIS2）-R	GAGCTCTTCTAGACATAAAAGTAAACATTAAATTATATAAACC
	PavMYB10.1-（PGADT7）-F	CATATGATGGCCAACAAGAAGATGGAGG
	PavMYB10.1-（PGADT7）-R	GGATCCGTCCTTCTGAACATTGGTACACTGTC

Fig. 1 Phenotype（A）,anthocyanin content（B）and relative expressions of PavMYB10.1 and PavRiant（C）in‘Tieton’sweet cherry fruit ** Indicate significant difference between S1（P < 0.01）.

Fig. 2 Phylogenetic tree analysis of GST proteins in different species

Fig. 3 Transient silence of PavRiant inhibits anthocyanin accumulation in fruit phenotype（A）,anthocyanin content（B） and relative expressions of PavRiant and anthocyanin related genes（C）in‘Tieton’sweet cherry fruit ** P < 0.01.

Fig. 4 PavMYB10.1 binds to the promoter of PavRiant A：Y1H assay showing the interaction between PavMYB10.1 and the PavRiant promoter;B：MBS and MRE sites in the PavRiant promoter;C：Electrophoretic mobility shift assay（EMSA）showing that PavMYB10.1 can bind to the MRE site of the PavRiant promoter. The“+”and “-”indicate the presence and absence of the probe and protein. represents the increasing concentration of competition and mutation probes.

Fig. 5 PavMYB10.1 activates the promoter activities of PavRiant **P < 0.01.

References 24

[1]	Allan A C, Hellens R P, Laing W A. 2008. MYB transcription factors that colour our fruit. Trends in Plant Science, 13:99-102. doi: 10.1016/j.tplants.2007.11.012 URL
[2]	Bai S L, Saito T, Honda C, Hatsuyama Y, Ito A, Moriguchi T. 2014. An apple B-box protein,MdCOL11,is involved in UV-B- and temperature-induced anthocyanin biosynthesis. Planta, 240 (5):1051-1062. doi: 10.1007/s00425-014-2129-8 URL
[3]	Cheng J, Liao L, Zhou H, Gu C, Wang L, Han Y P. 2015. A small indel mutation in an anthocyanin transporter causes variegated colouration of peach flowers. Journal of Experimental Botany, 66 (22):7227-7239. doi: 10.1093/jxb/erv419 pmid: 26357885
[4]	El-Sharkawy I, Liang D, Xu K. 2015. Transcriptome analysis of an apple(Malus × domestica)yellow fruit somatic mutation identifies a gene network module highly associated with anthocyanin and epigenetic regulation. Journal of Experimental Botany, 66 (22):7359-7376. doi: 10.1093/jxb/erv433 pmid: 26417021
[5]	Francisco R M, Regalado A, Ageorges A, Burla B J, Bassin B, Eisenach C, Zarrouk O, Vialet S, Marlin T, Chaves M M, Martinoia E, Nagy R. 2013. ABCC1,an ATP binding cassette protein from grape berry,transports anthocyanidin 3-O-glucosides. The Plant Cell, 25 (5):1840-1854. doi: 10.1105/tpc.112.102152 pmid: 23723325
[6]	Hu B, Zhao J T, Lai B, Qin Y H, Wang H C, Hu G B. 2016. LcGST4 is an anthocyanin-related glutathione S-transferase gene in Litchi chinensis Sonn. Plant Cell Reports, 35 (4):831-843. doi: 10.1007/s00299-015-1924-4 URL
[7]	Heppel S C, Jaffé F W, Takos A M, Schellmann S, Rausch T, Walker A R, Bogs J. 2013. Identification of key amino acids for the evolution of promoter target specificity of anthocyanin and proanthocyanidin regulating MYB factors. Plant Molecular Biology, 82:457-471. doi: 10.1007/s11103-013-0074-8 pmid: 23689818
[8]	Jaakola L. 2013. New insights into the regulation of anthocyanin biosynthesis in fruits. Trends in Plant Science, 18 (9):477-483. doi: 10.1016/j.tplants.2013.06.003 pmid: 23870661
[9]	Jiang S H, Chen M, He N B, Chen X L, Wang Nan, Sun Q G, Zhang T L, Xu H F, Fang H C, Wang Y C, Zhang Z Y, Wu S J, Chen X S. 2019. MdGSTF6,activated by MdMYB1,plays an essential role in anthocyanin accumulation in apple. Horticulture Research, 6:40. doi: 10.1038/s41438-019-0118-6 URL
[10]	Jin W M, Wang H, Li M F, Wang J, Yang Y, Zhang X M, Yan G H, Zhang H, Liu J S, Zhang K C. 2016. The R2R3 MYB transcription factor PavMYB10.1 involves in anthocyanin biosynthesis and determines fruit skin colour in sweet cherry(Prunus avium L.). Plant Biotechnology Journal, 14 (11):2120-2133. doi: 10.1111/pbi.12568 URL
[11]	Kim S H, Lee J R, Kim S R. 2006. Characterization of an apple anthocyanidin synthase gene in transgenic tobacco plants. Journal of Plant Biology, 49 (4):326-330. doi: 10.1007/BF03031164 URL
[12]	Kitamura S, Shikazono N, Tanaka A. 2004. TRANSPARENT TESTA 19 is involved in the accumulation of both anthocyanins and proanthocyanidins in Arabidopsis. The Plant Journal, 37 (1):104-114. doi: 10.1046/j.1365-313X.2003.01943.x URL
[13]	Luo H F, Dai C, Li Y P, Feng J, Liu Z C, Kang C Y. 2018. Reduced anthocyanins in petioles codes for a GST anthocyanin transporter that is essential for the foliage and fruit coloration in strawberry. Journal of Experimental Botany, 69 (10):2595-2608. doi: 10.1093/jxb/ery096 URL
[14]	Naing A H, Kim C K. 2018. Roles of R2R3-MYB transcription factors in transcriptional regulation of anthocyanin biosynthesis in horticultural plants. Plant Molecular Biology, 98:1-18. doi: 10.1007/s11103-018-0771-4 pmid: 30167900
[15]	Sun Y, Li H, Huang J R. 2012. Arabidopsis TT19functions as a carrier to transport anthocyanin from the cytosol to tonoplasts. Molecular Plant, 5 (2):387-400. doi: 10.1093/mp/ssr110 pmid: 22201047
[16]	Wei H R, Chen X, Zong X J, Shu H R, Gao D S, Liu Q Z. 2015. Comparative transcriptome analysis of genes involved in anthocyanin biosynthesis in the red and yellow fruits of sweet cherry(Prunus avium L.). PLoS ONE, 10 (3):e0121164. doi: 10.1371/journal.pone.0121164 URL
[17]	Wei K, Wang L Y, Zhang Y Z, Li R, Li H L, Wu L Y, Xu L Y, Zhang C C, Zhou X G, Cheng H, Edwards R. 2019. A coupled role for CsMYB75 and CsGSTF1 in anthocyanin hyperaccumulation in purple tea. The Plant Journal, 97 (5):825-840. doi: 10.1111/tpj.14161 URL
[18]	Xie D Y, Sharma S B, Paiva N L, Ferreira D, Dixon R A. 2003. Role of anthocyanidin reductase,encoded by BANYULS in plant flavonoid biosynthesis. Science, 299 (5605):396-399. doi: 10.1126/science.1078540 URL
[19]	Zeng J K, Li X, Xu Q, Chen J Y, Yin R X, Ferguson I B, Chen K S. 2015. EjAP2-1,an AP2/ERF gene,is a novel regulator of fruit lignification induced by chilling injury,via interaction with EjMYB transcription factors. Plant Biotechnology Journal, 13 (9):1325-1334. doi: 10.1111/pbi.12351 URL
[20]	Zhang Zhen, Li Chengzhiyu, Zhang Ya, Li Xi, Wang Shuo, Wang Xinyu, Chen Xueseng, Feng Shouqian. 2021. Inhibition of pear anthocyanin synthesis by the PcHB12gene in European pears. Acta Horticulturae Sinica, 48 (3):456-464. (in Chinese)
	张震, 李晨智宇, 张雅, 李玺, 王烁, 王鑫玉, 陈学森, 冯守千. 2021. 西洋梨PcHB12基因抑制果实花青苷的合成. 园艺学报, 48 (3):456-464.
[21]	Zhang Z, Tian C P, Zhang Y, Li C Z Y, Li X, Y Q, Wang S, Wang X Y, Chen X S, Feng S Q. 2020. Transcriptomic and metabolomic analysis provides insights into anthocyanin and procyanidin accumulation in pear. BMC Plant Biology, 20:129. doi: 10.1186/s12870-020-02344-0 pmid: 32220242
[22]	Zhao J. 2015. Flavonoid transport mechanisms:how to go,and with whom. Trends in Plant Science, 20 (9):576-585. doi: 10.1016/j.tplants.2015.06.007 URL
[23]	Zhao Y, Dong W Q, Zhu Y C, Allan A C, Wang K L, Xu C J. 2020. PpGST1,an anthocyanin-related glutathione S-transferase gene,is essential for fruit coloration in peach. Plant Biotechnology Journal, 18 (5):1284-1295. doi: 10.1111/pbi.13291 pmid: 31693790
[24]	Zhou H, Peng Q, Zhao J B, Owiti A, Ren F, Liao L, Wang L, Deng X B, Jiang Q, Han Y P. 2016. Multiple R2R3-MYB transcription factors involved in the regulation of anthocyanin accumulation in peach flower. Frontiers in Plant Science, 7:1557. pmid: 27818667

PavMYB10.1 Promotes Anthocyanin Accumulation by Positively Regulating PavRiant in Sweet Cherry

RichHTML

PDF

Knowledge

Abstract

Cite this article

share this article

Figures/Tables 6

References 24

Related Articles 15

Recommended Articles

Metrics

Comments

[1]	ZHANG Xiaoming, YAN Guohua, ZHOU Yu, WANG Jing, DUAN Xuwei, WU Chuanbao, and ZHANG Kaichun. A New Sweet Cherry Rootstock Cultivar‘Jingchun 2’ [J]. Acta Horticulturae Sinica, 2022, 49(S2): 31-32.
[2]	ZOU Xue, DING Fan, LIU Lifang, YU Hankaizong, CHEN Nianwei, and RAO Liping. A New Purple Potato Cultivar‘Mianziyu 1’ [J]. Acta Horticulturae Sinica, 2022, 49(S1): 93-94.
[3]	WANG Sha, ZHANG Xinhui, ZHAO Yujie, LI Bianbian, ZHAO Xueqing, SHEN Yu, DONG Jianmei, YUAN Zhaohe. Cloning and Functional Analysis of PgMYB111 Related to Anthocyanin Synthesis in Pomegranate [J]. Acta Horticulturae Sinica, 2022, 49(9): 1883-1894.
[4]	HUANG Ling, HU Xianmei, LIANG Zehui, WANG Yanping, CHAN Zhulong, XIANG Lin. Cloning and Function Identification of Anthocyanidin Synthase Gene TgANS in Tulipa gesneriana [J]. Acta Horticulturae Sinica, 2022, 49(9): 1935-1944.
[5]	LI Maofu, YANG Yuan, WANG Hua, FAN Youwei, SUN Pei, JIN Wanmei. Analysis the Function of R2R3 MYB Transcription Factor RhMYB113c on Regulating Anthocyanin Synthesis in Rosa hybrida [J]. Acta Horticulturae Sinica, 2022, 49(9): 1957-1966.
[6]	QIAN Jieyu, JIANG Lingli, ZHENG Gang, CHEN Jiahong, LAI Wuhao, XU Menghan, FU Jianxin, ZHANG Chao. Identification and Expression Analysis of MYB Transcription Factors Regulating the Anthocyanin Biosynthesis in Zinnia elegans and Function Research of ZeMYB9 [J]. Acta Horticulturae Sinica, 2022, 49(7): 1505-1518.
[7]	ZHANG Lugang, LU Qianqian, HE Qiong, XUE Yihua, MA Xiaomin, MA Shuai, NIE Shanshan, YANG Wenjing. Creation of Novel Germplasm of Purple-orange Heading Chinese Cabbage [J]. Acta Horticulturae Sinica, 2022, 49(7): 1582-1588.
[8]	CHEN Daozong, LIU Yi, SHEN Wenjie, ZHU Bo, TAN Chen. Identification and Analysis of PAP1/2 Homologous Genes in Brassica rapa,B. oleracea and B. napus [J]. Acta Horticulturae Sinica, 2022, 49(6): 1301-1312.
[9]	LI Xiaoming, YU Junchi, WANG Chunxia. Comparison of Growth and Secondary Metabolites of Purple and White Flower Dracocephalum moldavica Under Field,Greenhouse and Greenhouse Shading Conditions [J]. Acta Horticulturae Sinica, 2022, 49(6): 1363-1370.
[10]	DENG Jiao, SU Mengyue, LIU Xuelian, OU Kefang, HU Zhengrong, YANG Pingfang. Transcriptome Analysis Revealed the Formation Mechanism of Floral Color of Lotus‘Dasajin’with Bicolor Petal [J]. Acta Horticulturae Sinica, 2022, 49(2): 365-377.
[11]	WANG Zhiyu, CHANG Beibei, LIU Qi, CHENG Xiaofan, DU Xiaoyun, YU Xiaoli, SONG Laiqing, ZHAO Lingling. Study on Expression and Anthocyanin Accumulation of Solute Carrier Gene MdSLC35F2-like in Apple [J]. Acta Horticulturae Sinica, 2022, 49(11): 2293-2303.
[12]	SUN Wei, SUN Shiyu, CHEN Yiran, WANG Yuhan, ZHANG Yan, JU Zhigang, YI Yin. Cloning and Function Analysis of Chalcone Isomerase Gene RdCHI1 in Rhododendron delavayi [J]. Acta Horticulturae Sinica, 2022, 49(11): 2407-2418.
[13]	ZHANG Xiaoming, YAN Guohua, ZHOU Yu, WANG Jing, DUAN Xuwei, and ZHANG Kaichun, . A New Sweet Cherry Rootstock Cultivar‘Jingchun 1’ [J]. Acta Horticulturae Sinica, 2021, 48(S2): 2787-2788.
[14]	ZHANG Xiaoming, YAN Guohua, ZHOU Yu, WANG Jing, DUAN Xuwei, and ZHANG Kaichun, . A New Sweet Cherry Rootstock Cultivar‘Landing 3’ [J]. Acta Horticulturae Sinica, 2021, 48(S2): 2789-2790.
[15]	QI Xiliang, LIU Congli, SONG Lulu, LI Ming. Functional Analysis of Sucrose-phosphate Synthase Genes（SPS）in Sweet Cherry [J]. Acta Horticulturae Sinica, 2021, 48(8): 1446-1456.