石榴果色合成相关基因CHS和CHI的表达特性分析
|
摘要
花色苷是类黄酮家族中重要的一类次生代谢产物,对果实呈色起重要作用。CHS (查尔酮合成酶)和CHI (查尔酮异构酶)为花色苷合成提供了前体物质,是花色苷合成所不可或缺的。利用RT-PCR和RACE方法,本研究从石榴果皮中克隆了与花色苷合成相关的CHS基因和CHI基因的cDNA全长,同时采用qRT-PCR研究了这两个基因在三个不同色泽石榴品种‘红宝石’、‘水晶甜’、‘墨石榴’发育期内的表达模式,并分析了果皮花色苷含量变化与基因转录水平的关系。结果表明,石榴中CHS和CHI基因cDNA全长分别为1 197 bp和693 bp,分别编码398和230个氨基酸,命名为PgCHS和PgCHI,在GenBank中的登录号分别为KF841615和KF841616。在氨基酸水平上,Pg CHS与荔枝、葡萄、山竹等果树的同源性达到90%以上。Pg CHI与果树中龙眼、梨、美洲葡萄、桑树等同源性达到70%以上。qRT-PCR结果显示,CHS和CHI基因的表达模式随色泽发育期和品种不同而有差异。在‘红宝石’石榴中,该两个基因都有前期和后期两个表达高峰期;而‘水晶甜’石榴中这两个基因的表达高峰期均出现在中后期;‘墨石榴’发育初期时CHS和CHI的表达量最高,以后的表达量都较低。同一品种内,CHS和CHI的表达具有协同性,两者的协同性表达有利于花色苷及其他类黄酮相关产物的合成。3个品种中CHS和CHI基因的表达与花色苷的积累并不一致。
Anthocyanins, important secondary metabolites of flavonoids family, play important roles in fruit coloration. CHS(Chalcone synthase) and CHI(Chalcone isomerase) provide precursors for anthocyanin synthesis and are indispensable for anthocyanin synthesis. Using RT-PCR and RACE methods, we cloned DNA full-length of CHS gene and CHI gene related to anthocyanin synthesis from pomegranate fruit peels. At the same time, the expression patterns at development stages of these two genes in three different color pomegranate cultivars‘Hongbaoshi',‘Shuijingtian'and‘Moshiliu'were studied by qRT-PCR, and the relationship between anthocyanin content in pericarp and gene transcription level was analyzed. The results showed that the full-length of CHS gene and CHI gene cDNA in pomegranate were 1 197 bp and 693 bp respectively, coding 398 and 230 amino acids. They were denominated as PgCHS and PgCHI, and the accession numbers in GenBank were KF841615 and KF841616. At amino acid levels, Pg CHS possessed more than 90% identities with that in Litchi chinensis, Vitis vinifera, Garcinia mangostana, etc. Pg CHI had more than 70% identities with that in fruit trees such as Dimocarpus longan, Pyrus communis, Vitis labrusca, Morus alba. The results of qRT-PCR demonstrated that the expression patterns of CHS and CHI genes varied with color development stage and cultivars. In ‘Hongbaoshi'pomegranate, both of genes showed two expression peaks at the early and late stages; while in ‘Shuijingtian'cultivar, these two genes had expression peaks in the middle and later periods; For ‘Moshiliu'pomegranate, the expression level of CHS and CHI was the highest in the early developmental stages, and the expression level was lower afterwards. In the same cultivar, the profiles of CHS and CHI genes were synergetic, which was beneficial to the synthesis of anthocyanins and other flavonoids-related products. The expression of CHS and CHI genes in three pomegranate cultivars were inconsistent with the accumulation of total anthocyanins.
Anthocyanins, important secondary metabolites of flavonoids family, play important roles in fruit coloration. CHS(Chalcone synthase) and CHI(Chalcone isomerase) provide precursors for anthocyanin synthesis and are indispensable for anthocyanin synthesis. Using RT-PCR and RACE methods, we cloned DNA full-length of CHS gene and CHI gene related to anthocyanin synthesis from pomegranate fruit peels. At the same time, the expression patterns at development stages of these two genes in three different color pomegranate cultivars‘Hongbaoshi',‘Shuijingtian'and‘Moshiliu'were studied by qRT-PCR, and the relationship between anthocyanin content in pericarp and gene transcription level was analyzed. The results showed that the full-length of CHS gene and CHI gene cDNA in pomegranate were 1 197 bp and 693 bp respectively, coding 398 and 230 amino acids. They were denominated as PgCHS and PgCHI, and the accession numbers in GenBank were KF841615 and KF841616. At amino acid levels, Pg CHS possessed more than 90% identities with that in Litchi chinensis, Vitis vinifera, Garcinia mangostana, etc. Pg CHI had more than 70% identities with that in fruit trees such as Dimocarpus longan, Pyrus communis, Vitis labrusca, Morus alba. The results of qRT-PCR demonstrated that the expression patterns of CHS and CHI genes varied with color development stage and cultivars. In ‘Hongbaoshi'pomegranate, both of genes showed two expression peaks at the early and late stages; while in ‘Shuijingtian'cultivar, these two genes had expression peaks in the middle and later periods; For ‘Moshiliu'pomegranate, the expression level of CHS and CHI was the highest in the early developmental stages, and the expression level was lower afterwards. In the same cultivar, the profiles of CHS and CHI genes were synergetic, which was beneficial to the synthesis of anthocyanins and other flavonoids-related products. The expression of CHS and CHI genes in three pomegranate cultivars were inconsistent with the accumulation of total anthocyanins.
引文
Almeida J.R.M.,D'Amico E.,Preuss A.,Carbone F.,de Vos C.H.R.,Deiml B.,Mourgues F.,Perrotta G.,Fischer T.C.,Bovy A.G.,Martens S.,and Rosati C.,2007,Characterization of major enzymes and genes involved in flavonoid and proanthocyanidin biosynthesis during fruit development in strawberry(Fragaria×ananassa),Arch.Biochem.Biophys.,465(1):61-71
Ben-Simhon Z.,Judeinstein S.,Nadler-Hassar T.,Trainin T.,BarYa'akov I.,Borochov-Neori H.,and Holland D.,2011,Apomegranate(Punica granatum L.)WD40-repeat gene is a functional homologue of Arabidopsis TTG1 and is involved in the regulation of anthocyanin biosynthesis during pomegranate fruit development,Planta,234(5):865-881
Ben-Simhon Z.,Judeinstein S.,Trainin T.,Harel-Beja R.,BarY a'akov I.,Borochov-Neori H.,and Holland D.,2015,A'white'anthocyanin-less pomegranate(Punica granatum L.)caused by an insertion in the coding region of the leucoanthocyanidin dioxygenase(LDOX;ANS)gene,PLoS One,10(11):e0142777
Boss P.K.,Davies C.,and Robinson S.P.,1996,Analysis of the expression of anthocyanin pathway genes in developing Vitis vinifrera L.cv.Shiraz grape berries and the implications for pathway regulation,Plant Physiol.,111(4):1059-1066
Carbone F.,Preuss A.,De Vos R.C.,D'amico E.,Perrotta G.,Bovy A.G.,Martens S.,and Rosati C.,2009,Developmental,genetic and environmental factors affect the expression of flavonoid genes,enzymes and metabolites in strawberry fruits,Plan Cell Environ.,32(8):1117-1131
Dixon R.A.,and Steele C.L.,1999,Flavonoids and isoflavonoids a goldmine for metabolic engineering,Trends in Plant Sci.,4(10):394-400
Fischer U.A.,Carle R.,and Kammerer D.R.,2011,Identification and quantification of phenolic compounds from pomegranate(Punica granatum L.)peel,mesocarp,aril and differently produced juices by HPLC-DAD-ESI/MSn,Food Chem.,127(2):807-821
Gil M.I.,García-Viguera C.,Artés F.,and Tomás-Barberán F.A.1995,Changes in pomegranate juice pigmentation during ripening,J.Sci.Food.Agr.,68(1):77-81
Gil M.I.,Tomás-Barberán F.A.,Hess-Pierce B.,Holcroft D.M.and Kader A.A.,2000,Antioxidant activity of pomegranate juice and its relationship with phenolic composition and processing,J.Agr.Food Chem.,48(10):4581-4589
Hahlbrock K.,and Scheel D.,1989,Physiology and molecular bi ology of phenylpropanoid metabolism,Ann.Rev.Plan Biol.,(40):347-369
Holton T.A.,and Cornish E.C.,1995,Genetics and biochemistry of anthocyanin biosynthesis,Plant Cell,7(7):1071-1083
Jaakola L.,M??tt??K.,Pirttil?A.M.,T?rr?nen R.,K?renlampi S.and Hohtola A.,2002,Expression of genes involved in anthocyanin biosynthesis in relation to anthocyanin,proanthocyanidin,and flavonol levels during bilberry fruit development,Plant Physiol.,130(2):729-739
Jez J.M.,Bowman M.E.,Dixon R.A.,and Noel J.P.,2000,Structure and mechanism of the evolutionarily unique plant enzyme chalcone isomerse,Nature Structural Biology,7(9):786-791
Ju Z.G.,Liu C.L.,and Yuan Y.B.,1995,Activities of chalcone synthase and UDPGal:flavonid-3-o-glycosyltransferase in relation to anthocyanin synthesis in apple,Scientia Hortic.,63(3):175-185
Kondo S.,Hiraoka K.,Kobayashi S.,Honda C.,and Terahara N.,2002,Changes in the expression of anthocyanin biosynthetic genes during apple development,J.Am.Soc.Hortic.Sci.,,127(6):971-976
Livak K.J.,and Schmittgen T.D.,2001,Analysis of relative gene expression data using real-time quantitative PCR and the2-△△CTmethod,Methods,25(4):402-408
Mehrtens F.,Kranz H.,Bednarek P.,and Weisshaar B.,2005,The Arabidopsis transcription factor MYB12 is a flavonol-specific regulator of phenylpropanoid biosynthesis,Plant Physiol.,138(2):1083-1096
Noda Y.,Kaneyuki T.,Mori A.,and Packer L.,2002,Antioxidant activities of pomegranate fruit extract and its anthocyanidins:delphinidin,cyanidin,and pelargonidin,J.Agr.Food Chem.,50(1):166-171
Salvatierra A.,Pimentel P.,Moya-Leon M.A.,Caligari P.D.S.,and Herrera R.,2010,Comparison of transcriptional profiles of flavonoid genes and anthocyanin contens during fruit development of two botanical forms of Fragaria chiloensis ssp.chiloensis,Phytochemistry,71(16):1839-1847
Takos A.M.,Ubi B.E.,Robinson S.P.,and Walker A.R.,2006,Condensed tannin biosynthesis genes are regulated separately from other flavonoid biosynthesis genes in apple fruit skin,Plant Sci.,170(3):487-499
Tehranifar A.,Zarei M.,Nemati Z.,Esfandiyari B.,and Vazifesh enas M.R.,2010,Investigation of physico-chemical properties and antioxidant activity of twenty Iranian pomegranate(Punica granatum L.)cultivars,Scientia Hortic.,126(2):180-185
Tsuda T.,Yamaguchi M.,Honda C.,and Moriguchi T.,2004,Expression of anthocyanin biosynthesis genes in the skin of peach and nectarine fruit,J.Am.Soc.Hortic.Sci.,129(6):857-862
Van Elswijk D.A.,Schobel U.P.,Lansky E.P.,Irth H.,and van der Greef J.,2004,Rapid dereplication of estrogenic compounds in pomegranate(Punica granatum)using on-line biochemical detection coupled to mass spectrometry,Phytochemistry,65(2):233-241
Van Tunen A.J.,Koes R.E.,Spelt C.E.,van der Krol A.R.,Stuitje A.R.,and Mol J.N.,1988,Cloning of the 2 chalcone flavanone isomerase genes from Pentunia hybrida:coordinate,light-regulated and differential expression of flavonoid genes,EMBOJournal,7(5):1257-1263
Viuda-Martos M.,Fernández-López J.,and Pérez-魣lvarez J.A.2010,Pomegranate and its many functional components as related to human health:a review,Compr.Rev.Food Sci.F.9(6):635-654
Wang H.C.,Huang X.M.,Hu G.B.,and Huang H.B,2004,Stuidies on the relationship between anthocyanin biosynthesis and related enzymes in Litchi pericarp,Zhongguo Nongye Kexue(Scientia Agricultura Sinica),37(12):2028-2032(王惠聪,黄旭明,胡桂兵,黄辉白,2004,荔枝果皮花青苷合成与相关酶的关系研究,中国农业科学,37(12):2028-2032)
Winkel-shirley B.,2001,Flavonoid biosynthesis.A colorful mode for genetics,biochemistry,cell biology,and biotechnology,Plant Physiol.,126(2):485-493
Yao L.H.,Jiang Y.M.,Shi J.,Tomás-barberán F.A.,Datta N.Singanusong R.,and Chen S.S.,2004,Flavonoids in food and their health benefits,Plant Foods for Human Nutrition59(3):113-122
Yu X.N.,and Zhang Q.X.,2002,Anthocyanin in ornamental plan and color express,Linyekexue(Scientia Silvae Sinicae),38(3):147-153(于晓南,张启翔,2002,观赏植物的花色素苷与花色,林业科学,38(3):147-153)
Yuan Z.,Fang Y.,Zhang T.,Fei Z.,Han F.,Liu C.,Liu M.,Xiao W.,Zhang W.,Wu S.,Zhang M.,Ju Y.,Xu H.,Dai H.,Liu Y.,Chen Y.,Wang L.,Zhou J.,Guan D.,Yan M.,Xia Y.Huang X.,Liu D.,Wei H.,and Zheng H.,2018,The pomegranate(Punia granatum L.)genome provides insights into fruit quality and ovule developmental biology,Plant Biotechnol.J.,16(7):1363-1374
Yuan Z.,Yin Y.,Qu J.,Zhu L.,and Li Y.,2007,Populatioon genetic diversity in Chinese pomegranate(Punica granatum L.)cultivars revealed by fluorescent-AFLP markers,J.Genet Genomics,34(12):1061-1071
Yuan Z.H.,Yin Y.L.,Zhu L.Q.,Li Y.,and Hou L.F.,2008,Genetic diversity and relationship in pomegranate(Punica granatum L.)cultivars in Shandong revealed by fluorescent-AFLPmarkers,Yuanyi Xuebao(Acta Horticulturae Sinica),35(1):107-112(苑兆和,尹燕雷,朱丽琴,李云,侯乐峰,2008,山东石榴品种遗传多样性与亲缘关系的荧光AFLP分析,园艺学报,35(1):107-112)
Yuan Z.H.,ed.,2015,Chinese fruit tree science and practice pomegranate,Shaanxi Science and Technology Press,Xi'an,China,pp.26-42(苑兆和,主编,2015,中国果树科学与实践·石榴,陕西科学技术出版社,中国,西安,pp.26-42)
Zhang L.,Fu Q.,and Zhang Y.,2011,Composition of anthocyanins in pomegranate flowers and their antioxidant activity,Food Chemistry,127(4):1444-1449
Zhao X.,Yuan Z.,Fang Y.,Yin Y.,and Feng L.,2013,Characterization and evaluation of major anthocyanins in pomegranate(Punica granatum L.)peel of different cultivars and their development phases,Eur.Food Res.Technol.,236(1):109-117
Zhao X.,Yuan Z.,Fang Y.,Yin Y.,and Feng L.,2014,Flavonols and flavones changes in pomegranate(Punica granatum L.)fruit peel during fruit development,J.Agr.Sci.Technol.,16:1649-1659
Zhao X.,Yuan Z.,Feng L.,and Fang Y.,2015,Cloning and expression of anthocyanin biosynthetic genes in red and white pomegranate,J.Plant Res.,128(4):687-696
Zhou J.,Tao J.M.,Peng R.H.,Xiong A.S.,Cai B.,Xu J.T.,Jin X.F.,Zhang B.,Gao F.,Gao J.J.,Zhang Z.,and Yao Q.H.,2010,Cloning and expression analysis of CHS4 of Kyoho grape by RT-PCR.,Nanjin Nongye Daxue Xuebao(Journal of Nanjing Agricultural University),33(2):39-44(周军,陶建敏,彭日荷,熊爱生,蔡斌,徐锦涛,金晓芬,张斌,高峰,高建杰,章镇,姚泉洪,2010,巨峰葡萄查尔酮合成酶基因4的克隆及表达特性的RT-PCR分析,南京农业大学学报,33(2):39-44)
Ben-Simhon Z.,Judeinstein S.,Nadler-Hassar T.,Trainin T.,BarYa'akov I.,Borochov-Neori H.,and Holland D.,2011,Apomegranate(Punica granatum L.)WD40-repeat gene is a functional homologue of Arabidopsis TTG1 and is involved in the regulation of anthocyanin biosynthesis during pomegranate fruit development,Planta,234(5):865-881
Ben-Simhon Z.,Judeinstein S.,Trainin T.,Harel-Beja R.,BarY a'akov I.,Borochov-Neori H.,and Holland D.,2015,A'white'anthocyanin-less pomegranate(Punica granatum L.)caused by an insertion in the coding region of the leucoanthocyanidin dioxygenase(LDOX;ANS)gene,PLoS One,10(11):e0142777
Boss P.K.,Davies C.,and Robinson S.P.,1996,Analysis of the expression of anthocyanin pathway genes in developing Vitis vinifrera L.cv.Shiraz grape berries and the implications for pathway regulation,Plant Physiol.,111(4):1059-1066
Carbone F.,Preuss A.,De Vos R.C.,D'amico E.,Perrotta G.,Bovy A.G.,Martens S.,and Rosati C.,2009,Developmental,genetic and environmental factors affect the expression of flavonoid genes,enzymes and metabolites in strawberry fruits,Plan Cell Environ.,32(8):1117-1131
Dixon R.A.,and Steele C.L.,1999,Flavonoids and isoflavonoids a goldmine for metabolic engineering,Trends in Plant Sci.,4(10):394-400
Fischer U.A.,Carle R.,and Kammerer D.R.,2011,Identification and quantification of phenolic compounds from pomegranate(Punica granatum L.)peel,mesocarp,aril and differently produced juices by HPLC-DAD-ESI/MSn,Food Chem.,127(2):807-821
Gil M.I.,García-Viguera C.,Artés F.,and Tomás-Barberán F.A.1995,Changes in pomegranate juice pigmentation during ripening,J.Sci.Food.Agr.,68(1):77-81
Gil M.I.,Tomás-Barberán F.A.,Hess-Pierce B.,Holcroft D.M.and Kader A.A.,2000,Antioxidant activity of pomegranate juice and its relationship with phenolic composition and processing,J.Agr.Food Chem.,48(10):4581-4589
Hahlbrock K.,and Scheel D.,1989,Physiology and molecular bi ology of phenylpropanoid metabolism,Ann.Rev.Plan Biol.,(40):347-369
Holton T.A.,and Cornish E.C.,1995,Genetics and biochemistry of anthocyanin biosynthesis,Plant Cell,7(7):1071-1083
Jaakola L.,M??tt??K.,Pirttil?A.M.,T?rr?nen R.,K?renlampi S.and Hohtola A.,2002,Expression of genes involved in anthocyanin biosynthesis in relation to anthocyanin,proanthocyanidin,and flavonol levels during bilberry fruit development,Plant Physiol.,130(2):729-739
Jez J.M.,Bowman M.E.,Dixon R.A.,and Noel J.P.,2000,Structure and mechanism of the evolutionarily unique plant enzyme chalcone isomerse,Nature Structural Biology,7(9):786-791
Ju Z.G.,Liu C.L.,and Yuan Y.B.,1995,Activities of chalcone synthase and UDPGal:flavonid-3-o-glycosyltransferase in relation to anthocyanin synthesis in apple,Scientia Hortic.,63(3):175-185
Kondo S.,Hiraoka K.,Kobayashi S.,Honda C.,and Terahara N.,2002,Changes in the expression of anthocyanin biosynthetic genes during apple development,J.Am.Soc.Hortic.Sci.,,127(6):971-976
Livak K.J.,and Schmittgen T.D.,2001,Analysis of relative gene expression data using real-time quantitative PCR and the2-△△CTmethod,Methods,25(4):402-408
Mehrtens F.,Kranz H.,Bednarek P.,and Weisshaar B.,2005,The Arabidopsis transcription factor MYB12 is a flavonol-specific regulator of phenylpropanoid biosynthesis,Plant Physiol.,138(2):1083-1096
Noda Y.,Kaneyuki T.,Mori A.,and Packer L.,2002,Antioxidant activities of pomegranate fruit extract and its anthocyanidins:delphinidin,cyanidin,and pelargonidin,J.Agr.Food Chem.,50(1):166-171
Salvatierra A.,Pimentel P.,Moya-Leon M.A.,Caligari P.D.S.,and Herrera R.,2010,Comparison of transcriptional profiles of flavonoid genes and anthocyanin contens during fruit development of two botanical forms of Fragaria chiloensis ssp.chiloensis,Phytochemistry,71(16):1839-1847
Takos A.M.,Ubi B.E.,Robinson S.P.,and Walker A.R.,2006,Condensed tannin biosynthesis genes are regulated separately from other flavonoid biosynthesis genes in apple fruit skin,Plant Sci.,170(3):487-499
Tehranifar A.,Zarei M.,Nemati Z.,Esfandiyari B.,and Vazifesh enas M.R.,2010,Investigation of physico-chemical properties and antioxidant activity of twenty Iranian pomegranate(Punica granatum L.)cultivars,Scientia Hortic.,126(2):180-185
Tsuda T.,Yamaguchi M.,Honda C.,and Moriguchi T.,2004,Expression of anthocyanin biosynthesis genes in the skin of peach and nectarine fruit,J.Am.Soc.Hortic.Sci.,129(6):857-862
Van Elswijk D.A.,Schobel U.P.,Lansky E.P.,Irth H.,and van der Greef J.,2004,Rapid dereplication of estrogenic compounds in pomegranate(Punica granatum)using on-line biochemical detection coupled to mass spectrometry,Phytochemistry,65(2):233-241
Van Tunen A.J.,Koes R.E.,Spelt C.E.,van der Krol A.R.,Stuitje A.R.,and Mol J.N.,1988,Cloning of the 2 chalcone flavanone isomerase genes from Pentunia hybrida:coordinate,light-regulated and differential expression of flavonoid genes,EMBOJournal,7(5):1257-1263
Viuda-Martos M.,Fernández-López J.,and Pérez-魣lvarez J.A.2010,Pomegranate and its many functional components as related to human health:a review,Compr.Rev.Food Sci.F.9(6):635-654
Wang H.C.,Huang X.M.,Hu G.B.,and Huang H.B,2004,Stuidies on the relationship between anthocyanin biosynthesis and related enzymes in Litchi pericarp,Zhongguo Nongye Kexue(Scientia Agricultura Sinica),37(12):2028-2032(王惠聪,黄旭明,胡桂兵,黄辉白,2004,荔枝果皮花青苷合成与相关酶的关系研究,中国农业科学,37(12):2028-2032)
Winkel-shirley B.,2001,Flavonoid biosynthesis.A colorful mode for genetics,biochemistry,cell biology,and biotechnology,Plant Physiol.,126(2):485-493
Yao L.H.,Jiang Y.M.,Shi J.,Tomás-barberán F.A.,Datta N.Singanusong R.,and Chen S.S.,2004,Flavonoids in food and their health benefits,Plant Foods for Human Nutrition59(3):113-122
Yu X.N.,and Zhang Q.X.,2002,Anthocyanin in ornamental plan and color express,Linyekexue(Scientia Silvae Sinicae),38(3):147-153(于晓南,张启翔,2002,观赏植物的花色素苷与花色,林业科学,38(3):147-153)
Yuan Z.,Fang Y.,Zhang T.,Fei Z.,Han F.,Liu C.,Liu M.,Xiao W.,Zhang W.,Wu S.,Zhang M.,Ju Y.,Xu H.,Dai H.,Liu Y.,Chen Y.,Wang L.,Zhou J.,Guan D.,Yan M.,Xia Y.Huang X.,Liu D.,Wei H.,and Zheng H.,2018,The pomegranate(Punia granatum L.)genome provides insights into fruit quality and ovule developmental biology,Plant Biotechnol.J.,16(7):1363-1374
Yuan Z.,Yin Y.,Qu J.,Zhu L.,and Li Y.,2007,Populatioon genetic diversity in Chinese pomegranate(Punica granatum L.)cultivars revealed by fluorescent-AFLP markers,J.Genet Genomics,34(12):1061-1071
Yuan Z.H.,Yin Y.L.,Zhu L.Q.,Li Y.,and Hou L.F.,2008,Genetic diversity and relationship in pomegranate(Punica granatum L.)cultivars in Shandong revealed by fluorescent-AFLPmarkers,Yuanyi Xuebao(Acta Horticulturae Sinica),35(1):107-112(苑兆和,尹燕雷,朱丽琴,李云,侯乐峰,2008,山东石榴品种遗传多样性与亲缘关系的荧光AFLP分析,园艺学报,35(1):107-112)
Yuan Z.H.,ed.,2015,Chinese fruit tree science and practice pomegranate,Shaanxi Science and Technology Press,Xi'an,China,pp.26-42(苑兆和,主编,2015,中国果树科学与实践·石榴,陕西科学技术出版社,中国,西安,pp.26-42)
Zhang L.,Fu Q.,and Zhang Y.,2011,Composition of anthocyanins in pomegranate flowers and their antioxidant activity,Food Chemistry,127(4):1444-1449
Zhao X.,Yuan Z.,Fang Y.,Yin Y.,and Feng L.,2013,Characterization and evaluation of major anthocyanins in pomegranate(Punica granatum L.)peel of different cultivars and their development phases,Eur.Food Res.Technol.,236(1):109-117
Zhao X.,Yuan Z.,Fang Y.,Yin Y.,and Feng L.,2014,Flavonols and flavones changes in pomegranate(Punica granatum L.)fruit peel during fruit development,J.Agr.Sci.Technol.,16:1649-1659
Zhao X.,Yuan Z.,Feng L.,and Fang Y.,2015,Cloning and expression of anthocyanin biosynthetic genes in red and white pomegranate,J.Plant Res.,128(4):687-696
Zhou J.,Tao J.M.,Peng R.H.,Xiong A.S.,Cai B.,Xu J.T.,Jin X.F.,Zhang B.,Gao F.,Gao J.J.,Zhang Z.,and Yao Q.H.,2010,Cloning and expression analysis of CHS4 of Kyoho grape by RT-PCR.,Nanjin Nongye Daxue Xuebao(Journal of Nanjing Agricultural University),33(2):39-44(周军,陶建敏,彭日荷,熊爱生,蔡斌,徐锦涛,金晓芬,张斌,高峰,高建杰,章镇,姚泉洪,2010,巨峰葡萄查尔酮合成酶基因4的克隆及表达特性的RT-PCR分析,南京农业大学学报,33(2):39-44)