‘秋姬李’PsMYB18基因克隆与功能分析Molecular cloning and function analysis of PsMYB18 in 'Akihime' plum (Prunus salicina Lindl.)
方智振;姜翠翠;周丹蓉;潘少霖;林炎娟;叶新福;
摘要(Abstract):
【目的】克隆在采后‘秋姬李’果皮积累花色苷过程中高表达的MYB抑制因子PsMYB18基因,研究其序列特征、表达特点与功能。【方法】以‘秋姬李’为试材,采用qRT-PCR分析不同温度和光照处理条件下‘秋姬李’果皮中PsMYB18基因的转录水平,采用RT-PCR克隆PsMYB18基因,并通过烟草叶片瞬时表达试验分析PsMYB18的功能。【结果】q RT-PCR分析表明20℃和光照处理可促进‘秋姬李’果皮PsMYB18基因表达。PsMYB18基因的开放阅读框(ORF)为702 bp,编码233个氨基酸的蛋白。进化树分析表明PsMYB18与其他植物的花色苷合成抑制因子亲缘关系较近。序列比对结果表明其具有保守的R2R3结构域和抑制基序C1和C2。烟草瞬时表达试验表明,PsMYB18可抑制正调控因子PsMYB10.1和PsbHLH3的花色苷合成诱导功能。【结论】‘秋姬李’PsMYB18为花色苷合成抑制因子。
关键词(KeyWords): ‘秋姬李’;PsMYB18;花色苷合成抑制因子
基金项目(Foundation): 国家自然科学基金(31801916);; 福建省省属公益类科研院所基本科研专项(2018R1013-1);; 福建农业科学院青年英才专项(YC2015-9);; 福建省农业科学院创新团队(STIT2017-1-4)
作者(Author): 方智振;姜翠翠;周丹蓉;潘少霖;林炎娟;叶新福;
Email:
DOI: 10.13925/j.cnki.gsxb.20190080
参考文献(References):
- [1]刘晓芬,李方,殷学仁,徐昌杰,陈昆松.花青苷生物合成转录调控研究进展[J].园艺学报,2013,40(11):2295-2306.LIU Xiaofen,LI Fang,YIN Xueren,XU Changjie,CHEN Kunsong. Recent advances in the transcriptional regulation of anthocyanin biosynthesis[J]. Acta Horticulturae Sinica,2013,40(11):2295-2306.
- [2] CHENG Y,LIU L,YUAN C,GUAN J. Molecular characterization of ethylene-regulated anthocyanin biosynthesis in plums during fruit ripening[J]. Plant Molecular Biology Reporter,2016,34(4):777-785.
- [3] BAI S,SUN Y,QIAN M,YANG F,NI J,TAO R,LI L,SHU Q,ZHANG D,TENG Y. Transcriptome analysis of bagging-treated red Chinese sand pear peels reveals light-responsive pathway functions in anthocyanin accumulation[J]. Scientific Reports,2017,7(1):63.
- [4] TAO R,BAI S,NI J,YANG Q,ZHAO Y,TENG Y. The blue light signal transduction pathway is involved in anthocyanin accumulation in‘Red Zaosu’pear[J]. Planta,2018,248(1):37-48.
- [5] WANG N,QU C,JIANG S,CHEN Z,XU H,FANG H,SU M,ZHANG J,WANG Y,LIU W,ZHANG Z,LU N,CHEN X. The proanthocyanidin-specific transcription factor MdMYBPA1 initiates anthocyanin synthesis under low-temperature conditions in red-fleshed apples[J]. The Plant Journal,2018,96(1):39-55.
- [6] PAN H,WANG R,LI L,WANG J,CAO J,JIANG W. Manipulation of ripening progress of different plum cultivars during shelf life by post-storage treatments with ethylene and 1-methylcyclopropene[J]. Scientia Horticulturae,2016,198:176-182.
- [7] ZHAO J,DIXON R A. The‘ins’and‘outs’of flavonoid transport[J]. Trends in Plant Science,2010,15(2):72-80.
- [8] ALLAN A C,HELLENS R P,LAING W A. MYB transcription factors that colour our fruit[J]. Trends in Plant Science,2008,13(3):99-102.
- [9] ESPLEY R V,HELLENS R P,PUTTERILL J,STEVENSON D E,KUTTY-AMMA S,ALLAN A C. Red colouration in apple fruit is due to the activity of the MYB transcription factor,MdMYB10[J].The Plant Journal,2007,49(3):414-427.
- [10] ZHANG Y,FANG Z,YE X,PAN S. Identification of candidate genes involved in anthocyanin accumulation in the peel of jaboticaba(Myrciaria cauliflora)fruits by transcriptomic analysis[J]. Gene,2018,676:202-213.
- [11] WANG N,LIU W,ZHANG T,JIANG S,XU H,WANG Y,ZHANG Z,WANG C,CHEN X. Transcriptomic analysis of redfleshed apples reveals the novel role of MdWRKY11 in flavonoid and anthocyanin biosynthesis[J]. Journal of Agricultural and Food Chemistry,2018,66(27):7076-7086.
- [12] LIU C C,CHI C,JIN L J,ZHU J,YU J Q,ZHOU Y H. The bZip transcription factor HY5 mediates CRY1a-induced anthocyanin biosynthesis in tomato[J]. Plant,Cell and Environment,2018,41(8):1762-1775.
- [13] AN J P,YAO J F,XU R R,YOU C X,WANG X F,HAO Y J.Apple bZIP transcription factor MdbZIP44 regulates ABA-promoted anthocyanin accumulation[J]. Plant,Cell and Environment,2018,41(11):2678-2692.
- [14] ZHOU H,WANG L K,WANG H,GU C,DARE A P,ESPLEY R V,HE H,ALLAN A C,HAN Y. Molecular genetics of bloodfleshed peach reveals activation of anthocyanin biosynthesis by NAC transcription factors[J]. The Plant Journal,2015,82(1):105-121.
- [15] PREZ-DAZ J,PREZ-DAZ J,MADRID-ESPINOZA J,GONZ LEZ-VILLANUEVA E,MORENO Y,RUIZ-LARA S. New member of the R2R3-MYB transcription factors family in grapevine suppresses the anthocyanin accumulation in the flowers of transgenic tobacco[J]. Plant Molecular Biology,2015,90(1):63-76.
- [16] HUANG Y F,VIALET S,GUIRAUD JL,TORREGROSA L,BERTRAND Y,CHEYNIER V,THIS P,TERRIER N. A negative MYB regulator of proanthocyanidin accumulation,identified through expression quantitative locus mapping in the grape berry[J]. New Phytologist,2014,201(3):795-809.
- [17] ZHOU H,LIN-WANG K,WANG F,ESPLEY R V,REN F,ZHAO J,OGUTU C,HE H,JIANG Q,ALLAN A C,HAN Y.Activator-type R2R3-MYB genes induce a repressor-type R2R3-MYB gene to balance anthocyanin and proanthocyanidin accumulation[J]. New Phytologist,2019,221(4):1919-1934.
- [18] KOES R,VERWEIJ W,QUATTROCCHIO F. Flavonoids:a colorful model for the regulation and evolution of biochemical pathways[J]. Trends in Plant Science,2005,10(5):236-242.
- [19] ZHANG X Y,ZHANG S L,LUO J,YE Z W,GAO Q H,DU J H,WANG X Q. Developmental changes in anthocyanin biosynthesis in plum skins and the effects of ethephon[J]. The Journal of Horticultural Science and Biotechnology,2009,84(5):495-498.
- [20] JUN J H,LIU C,XIAO X,DIXON R A. The transcriptional repressor MYB2 regulates both spatial and temporal patterns of proanthocyandin and anthocyanin pigmentation in Medicago truncatula[J]. The Plant Cell,2015,27(10):2860-2879.
- [21] TOMINAGA R,IWATA M,OKADA K,WADA T. Functional analysis of the epidermal-specific MYB genes CAPRICE and WEREWOLF in Arabidopsis[J]. The Plant Cell,2007,19(7):2264-2277.
- [22] WANG S,HUBBARD L,CHANG Y,GUO J,SCHIEFELBEIN J,CHEN J G. Comprehensive analysis of single-repeat R3 MYB proteins in epidermal cell patterning and their transcriptional regulation in Arabidopsis[J]. BMC Plant Biology,2008,8(1):81.
- [23] ZHU H F,FITZSIMMONS K,KHANDELWAL A,KRANZ R G. CPC,a single-repeat R3 MYB,is a negative regulator of anthocyanin biosynthesis in Arabidopsis[J]. Molecular Plant,2009,2(4):790-802.
- [24] MATSUI K,UMEMURA Y,OHME-TAKAGI M. AtMYBL2,a protein with a single MYB domain,acts as a negative regulator of anthocyanin biosynthesis in Arabidopsis[J]. The Plant Journal,2008,55(6):954-967.
- [25] AHARONI A,DE VOS C H,WEIN M,SUN Z,GRECO R,KROON A,MOL J N,O'CONNELL A P. The strawberry FaMYB1 transcription factor suppresses anthocyanin and flavonol accumulation in transgenic tobacco[J]. The Plant Journal,2001,28(3):319-332.
- [26] HUANG D,WANG X,TANG Z,YUAN Y,XU Y,HE J,JIANG X,PENG S-A,LI L,BUTELLI E,DENG X,XU Q. Subfunctionalization of the Ruby2–Ruby1 gene cluster during the domestication of citrus[J]. Nature Plants,2018,4(11):930-941.
- [27] JIN H,COMINELLI E,BAILEY P,PARR A,MEHRTENS F,JONES J,TONELLI C,WEISSHAAR B,MARTIN C. Transcriptional repression by AtMYB4 controls production of UV‐protecting sunscreens in Arabidopsis[J]. The EMBO Journal,2000,19(22):6150-6161.
- [28] YOSHIDA K,MA D,CONSTABEL C P. The MYB182 protein downregulates proanthocyanidin and anthocyanin biosynthesis in poplar by repressing both structural and regulatory flavonoid genes[J]. Plant Physiology,2015,167(3):693-710.
- [29] KRANZ H D,DENEKAMP M,GRECO R,JIN H,LEYVA A,MEISSNER R C,PETRONI K,URZAINQUI A,BEVAN M,MARTIN C,SMEEKENS S,TONELLI C,PAZ-ARES J,WEISSHAAR B. Towards functional characterisation of the members of the R2R3-MYB gene family from Arabidopsis thaliana[J].The Plant Journal,1998,16(2):263-276.
- [30] LIN-WANG K,BOLITHO K,GRAFTON K,KORTSTEE A,KARUNAIRETNAM S,MCGHIE T,ESPLEY R,HELLENS R,ALLAN A. An R2R3 MYB transcription factor associated with regulation of the anthocyanin biosynthetic pathway in Rosaceae[J]. BMC Plant Biology,2010,10(1):50.
- [31] GU C,LIAO L,ZHOU H,WANG L,DENG X,HAN Y. Constitutive activation of an anthocyanin regulatory gene PcMYB10.6is related to red coloration in purple-foliage plum[J]. PLoS ONE,2015,10(8):e0135159.
- [32] FANG Z Z,ZHOU D R,YE X F,JIANG C C,PAN S L. Identification of candidate anthocyanin-related genes by transcriptomic analysis of‘Furongli’plum(Prunus salicina Lindl.)during fruit ripening using RNA-Seq[J]. Frontiers in Plant Science,2016,7:1338.
- [33] FANG Z Z,LAI C C,ZHANG Y L,LIN Y L,LAI Z X. Identification of a PTC-containing DlRan transcript and its differential expression during somatic embryogenesis in Dimocarpus longan[J]. Gene,2013,529(1):37-44.
- [34] LIN-WANG K,MICHELETTI D,PALMER J,VOLZ R,LOZANO L,ESPLEY R,HELLENS R P,CHAGNèD,ROWAN D D,TROGGIO M,IGLESIAS I,ALLAN A C. High temperature reduces apple fruit colour via modulation of the anthocyanin regulatory complex[J]. Plant,Cell and Environment,2011,34(7):1176-1190.
- [35] DUBOS C,STRACKE R,GROTEWOLD E,WEISSHAAR B,MARTIN C,LEPINIEC L. MYB transcription factors in Arabidopsis[J]. Trends in Plant Science,2010,15(10):573-581.
- [36] CAVALLINI E,MATUS J T,FINEZZO L,ZENONI S,LOYOLA R,GUZZO F,SCHLECHTER R,AGEORGES A,ARCE-JOHNSON P,TORNIELLI G B. The phenylpropanoid pathway is controlled at different branches by a set of R2R3-MYB C2 repressors in grapevine[J]. Plant Physiology,2015,167(4):1448-1470.
- [37] DUBOS C,LEGOURRIEREC J,BAUDRY A,HUEP G,LANET E,DEBEAUJON I,ROUTABOUL J M,ALBORESI A,WEISSHAAR B,LEPINIEC L. MYBL2 is a new regulator of flavonoid biosynthesis in Arabidopsis thaliana[J].The Plant Journal,2008,55(6):940-953.
- [38] ALBERT N W,LEWIS D H,ZHANG H,SCHWINN K E,JAMESON P E,DAVIES K M. Members of an R2R3-MYB transcription factor family in Petunia are developmentally and environmentally regulated to control complex floral and vegetative pigmentation patterning[J]. The Plant Journal,2011,65(5):771-784.
- [39] P REZ-D AZ J R,P REZ-DAZ J,MADRID-ESPINOZA J,GONZ LEZ-VILLANUEVA E,MORENO Y,RUIZ-LARA S.New member of the R2R3-MYB transcription factors family in grapevine suppresses the anthocyanin accumulation in the flowers of transgenic tobacco[J]. Plant Molecular Biology,2016,90(1):63-76.
- [40] ALBERT N W. Subspecialization of R2R3-MYB repressors for anthocyanin and proanthocyanidin regulation in forage legumes[J]. Frontiers Plant Science,2015,6:1165.
- [41] MA D,REICHELT M,YOSHIDA K,GERSHENZON J,CONSTABEL C P. Two R2R3-MYB proteins are broad repressors of flavonoid and phenylpropanoid metabolism in poplar[J]. The Plant Journal,2018,96(5):949-965.