桃肉质及粘离核性状形成及其相关基因的表达分析
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摘要
以大久保、08-9-106和08-9-107为试材,分析了果实成熟后期中果皮果肉硬度变化、粘离核性状的形成以及中果皮和近核处果肉的乙烯释放和相关基因表达的变化。结果表明,在果实中果皮中,大久保在成熟期果肉硬度下降快,乙烯释放明显,同期PpPG、PpACS1、PpYUC11相对表达量呈先升后降趋势;而08-9-106果肉硬度下降较快,但没有明显的乙烯释放和乙烯合成相关基因(PpACS1和PpYUC11)的表达;08-9-107在成熟后期硬度下降不明显,没有检测到明显的乙烯释放和乙烯合成相关基因的表达。在近核处果肉中,大久保的果实表现为离核性状,同期检测到明显的乙烯释放和相关合成基因(主要为PpACS1)的表达;而08-9-106的果实也表现为离核,但乙烯释放和基因表达与中果皮类似,均无明显检出;08-9-107表现为粘核,虽然乙烯释放量和PpACS1表达较低,但PpACO1和PpYUC11却有一定程度的表达。综上,大久保属于响应乙烯的离核溶质类型,08-9-106属于不响应乙烯的离核溶质类型,08-9-107属于响应乙烯的粘核硬质类型。桃果肉硬度的变化及粘离核性状的形成与PpPG基因的表达相关,PpACS1而非PpACO1是3份桃种质中调控乙烯含量的关键基因。
The fruits of peach varieties Okubo,08-9-106 and 08-9-107 were used as materials to analyze the fruit mesocarp firmness,flesh adhension,ethylene content,and related gene expression during the late fruit development period. The results showed that in Okubo,the flesh firmness decreased rapidly,and the accumulation of ethylene increased greatly,while the expression of PpPG,PpACS1,and PpYUC11 increased first and then decreased. In peach 08-9-106,the flesh firmness decreased rapidly,but there were no significant increase of ethylene and related gene(PpACS1 and PpYUC11)expression. Compared with Okubo and 08-9-106,08-9-107 had no significant increase in ethylene and gene expression,which might result in the fruit firmness. Based on the flesh near endocarp,the Okubo was characterized as freestone,with higher ethylene content and increased gene expression obviously(mainly PpACS1). 08-9-106 was also classified as freestone peach,which had no obvious ehtylene release and gene expression activation. 08-9-107 was clingstone,and exhibited the similar characters with 08-9-106,except its higher gene expression of PpACO1 and PpYUC11. The above results suggested that Okubo and 08-9-107 were freestone-melting type and clingstone stony-hard type,respectively,which responded to ethylene. 08-9-106 was freestone melting flesh phenotype,with no response to ethylene. The changes of flesh firmness and the formation of adhesion trait in peach fruits were both related to gene expression of PpPG. PpACS1(not the PpACO1)was the most essential gene in regulating the ethylene accumulation of the three peach germplasms.
The fruits of peach varieties Okubo,08-9-106 and 08-9-107 were used as materials to analyze the fruit mesocarp firmness,flesh adhension,ethylene content,and related gene expression during the late fruit development period. The results showed that in Okubo,the flesh firmness decreased rapidly,and the accumulation of ethylene increased greatly,while the expression of PpPG,PpACS1,and PpYUC11 increased first and then decreased. In peach 08-9-106,the flesh firmness decreased rapidly,but there were no significant increase of ethylene and related gene(PpACS1 and PpYUC11)expression. Compared with Okubo and 08-9-106,08-9-107 had no significant increase in ethylene and gene expression,which might result in the fruit firmness. Based on the flesh near endocarp,the Okubo was characterized as freestone,with higher ethylene content and increased gene expression obviously(mainly PpACS1). 08-9-106 was also classified as freestone peach,which had no obvious ehtylene release and gene expression activation. 08-9-107 was clingstone,and exhibited the similar characters with 08-9-106,except its higher gene expression of PpACO1 and PpYUC11. The above results suggested that Okubo and 08-9-107 were freestone-melting type and clingstone stony-hard type,respectively,which responded to ethylene. 08-9-106 was freestone melting flesh phenotype,with no response to ethylene. The changes of flesh firmness and the formation of adhesion trait in peach fruits were both related to gene expression of PpPG. PpACS1(not the PpACO1)was the most essential gene in regulating the ethylene accumulation of the three peach germplasms.
引文
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[10] Morgutti S,Negrini N,Nocito F F,Ghiani A,Bassi D,Cocucci M.Changes in endopolygalacturonase levels and characterization of a putative endo-PpPG gene during fruit softening in peach genotypes with nonmelting and melting flesh fruit phenotypes[J].New Phytologist,2010,171(2):315-328.doi:10.1111/j.1469-8137.2006.01763.x.
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[12] Bapat V A,Trivedi P K,Ghosh A,Sane V A,Thumballi G R,Nath P.Ripening of fleshy fruit:molecular insight and the role of ethylene[J].Biotechnology Advances,2010,28(1):94-107.doi:10.1016/j.biotechadv.2009.10.002.
[13] Haji T,Yaegaki H,Yamaguchi M.Changes in ethylene production and flesh firmness of melting,nonmelting and stony hard peaches after harvest[J].Journal of the Japanese Society for Horticultural Science,2001,70:458-459.doi:10.2503/jjshs.70.458.
[14] Goffreda J C.Stony hard gene of peach alters ethylene biosynthesis,respirationand oyher ripening related gharacteristic[J].HortScience,1992,27(6):610-610.
[15] Tsunashima Y,Owino W,Kubo Y,Inaba A,Mathooko F.Regulation of genes encoding ethylene biosynthetic enzymes in peach(Prunus persica L.)fruit by carbon dioxide and 1-methylcyclopropene[J].Postharvest Biology & Technology,2001,21(3):265-281.doi:10.1016/S0925-5214(00)00158-7.
[16] Tatsuki M,Haji T,Yamaguchi M.The involvement of 1-aminocyclopropane-1-carboxylic acid synthase isogene,PpACS1,in peach fruit softening[J].Journal of Experimental Botany,2006,57(6):1281-1289.doi:10.1093/jxb/erj097.
[17] Ohmiya A,Haji T.Promotion of ethylene biosynthesis in peach mesocarp discs by auxin[J].Plant Growth Regulation,2002,36(3):209-214.doi:10.1023/a:1016526517435.
[18] Pan L,Zeng W F,Niu L,Lu Z H,Liu H,Cui G C,Zhu Y Q,Chu J F,Li W P,Fang W C,Cai Z G,Li G H,Wang Z Q.PpYUC11,a strong candidate gene for the stony hard phenotype in peach(Prunus persica L.Batsch),participates in IAA biosynthesis during fruit ripening[J].Journal of Experimental Botany,2015,66(22):7031-7044.doi:10.1093/jxb/erv400.
[19] Marco C,Daniela G,Angelo C,Remo C,Stefano G,Laura R,Alessandro L,Daniele B.Integrative genomics approaches validate PpYUC11-like as candidate gene for the stony hard trait in peach (P.persica L.Batsch)[J].BMC Plant Biology,2018,18(1):88.doi:10.1186/s12870-018-1293-6.
[20] Kenneth J L,Thomas D S.Analysis of relative gene expression data using real-time quantitative PCR and the 2-ΔΔCT,method[J].Methods-A Companion To Methods in Enzymology,2001,25(4):402-408.doi:10.1006/meth.2001.1262.
[21] Mathooko F M,Tsunashima Y,Owino W Z O,Kubo Y,Inaba A.Regulation of genes encoding ethylene biosynthetic enzymes in peach(Prunus persica L.)fruit by carbon dioxide and 1-methylcyclopropene[J].Postharvest Biology and Technology,2001,21(3):265-281.doi:10.1016/S0925-5214(00)00158-7.
[22] 杨勇,马瑞娟,张斌斌,宋志忠,张春华,郭绍雷.不同溶质桃果实的软化与乙烯合成相关基因的差异表达[J].园艺学报,2015,42(10):1869-1878.doi:10.16420/j.issn.0513-353x.2015-0126.Yang Y,Ma R J,Zhang B B,Song Z Z,Zhang C H,Guo S L.Differential expression analysis in fruit softening and ethylene biosynthetic pathways in peaches of different flesh textures[J].Acta Horticulturae Sinica,2015,42(10):1869-1878.
[23] Ghiani A,Baldin F,Morgutti S,Negrini N,Nocito F F,Mignani I,Bassi D,Cocucci M.Changes in the expression of genes involved in ethylene signalling in peach fruit with different flesh texture and softening patterns[J].Acta Horticulturae,2010,884:125-131.doi:10.17660/ActaHortic.2010.884.13.
[24] Tatsuki M,Haji T,Yamaguchi M.The involvement of 1-aminocyclopropane-1-carboxylic acid synthase isogene,PpACS1,in peach fruit softening[J].Journal of Experimental Botany,2006,57(6):1281-1289.doi:10.1093/jxb/erj097.
[25] Liu M C,Pirrello J,Chervin C,Roustan J P,Bouzayen M.Ethylene control of fruit ripening:Revisiting the complex network of transcriptional regulation[J].Plant Physiology,2015,169(4):2380.doi:10.1104/pp.15.01361.
[26] 苏素香,赵彩平,曹丽军,李金金,李芳,韩明玉.两种不同耐贮性桃果实采后乙烯合成和果实软化相关基因表达的差异[J].农业生物技术学报,2015,23(4):450-458.doi:10.3969/j.issn.1674-7968.2015.04.004.Su S X,Zhao C P,Cao L J,Li J J,Li F,Han M Y.The difference of ethylene biosynthesis and expression of softening-related genes between two peach(Prunus persica)cultivars with different storage property after harvest[J].Journal of Agricultural Biotechnology,2015,23(4):450-458.
[27] 曾文芳,丁义峰,潘磊,王小贝,牛良,鲁振华,崔国朝,王志强.桃硬质性状可能源于PpYUC11基因启动子区域CACTA型转座子的插入[J].果树学报,2017,34(10):1239-1248.doi:10.13925/j.cnki.gsxb.20170060.Zeng W F,Ding Y F,Pan L,Wang X B,Niu L,Lu Z H,Cui G C,Wang Z Q.A CACTA transposable element in a PpYUC11 gene promoter is associated with the stony hard phenotype in peach[J].Journal of Fruit Science,2017,34(10):1239-1248.
[28] Tatsuki M,Nakajima N,Fujii H,Shimada T,Nakano M,Hayashi K,Hayama H,Yoshioka H,Nakamura Y.Increased levels of IAA are required for system 2 ethylene synthesis causing fruit softening in peach(Prunus persica L.Batsch)[J].Journal of Experimental Botany,2013,64(4):1049-1059.doi:10.1093/jxb/ers381.
[2] Yoshida M.Genetical studies on the fruit quality of peach varieties.Ⅲ.Texture and keeping quality[J].Bulletin of the Fruit Tree Research Station 3,1976,3:1-16.
[3] Bailey J S,French A P.The inheritance of certain characters in the peach[J].Proc Amer Soc Hort Sci,1932,29:127-130.
[4] Fry S C.Primary cell wall metabolism:Tracking the careers of wall polymers in living plant cells[J].New Phytologist,2004,161(3):641-675.doi:10.1111/j.1469-8137.2004.00980.x.
[5] Brummell D A.Cell wall disassembly in ripening fruit[J].Functional Plant Biology,2006,33(2):103-119.doi:10.1071/FP05234.
[6] Yoshioka H,Hayama H,Tatsuki M,Nakamura Y.Cell wall modifications during softening in melting type peach "Akatsuki" and non-melting type peach "Mochizuki"[J].Postharvest Biology & Technology,2011,60(2):100-110.doi:10.1016/j.postharvbio.2010.12.013.
[7] Trainotti L,Zanin D,Casadoro G.A cell wall oriented genomic approach reveals a new and unexpected complexity of the softening in peaches[J].Journal of Experimental Botany,2003,54(389):1821-1832.doi:10.1093/jxb/erg198.
[8] Ghiani A,Onelli E,Aina R,Cocucci M,Citterio S.A comparative study of melting and non-melting flesh peach cultivars reveals that during fruit ripening endo-polygalacturonase(endo-PpPG)is mainly involved in pericarp textural change,not in firmness reduction[J].Journal of Experimental Botany,2011,62(11):4043-4054.doi:10.1093/jxb/err109.
[9] Peace C P,Callahan A,Ogundiwin E A,Potter D,Gradziel T M,Bliss F A,Crisosto C H.Endopolygalacturonase genotypic variation in Prunus[J].Acta Horticulturae,2007,738:639-646.doi:10.17660/ActaHortic.2007.738.83.
[10] Morgutti S,Negrini N,Nocito F F,Ghiani A,Bassi D,Cocucci M.Changes in endopolygalacturonase levels and characterization of a putative endo-PpPG gene during fruit softening in peach genotypes with nonmelting and melting flesh fruit phenotypes[J].New Phytologist,2010,171(2):315-328.doi:10.1111/j.1469-8137.2006.01763.x.
[11] Mattoo A K,Suttle J C.The plant hormone ethylene[M].Boca Raton,FL:CRC Press,1991:293-314.doi:10.1007/978-1-4615-0675-1_1.
[12] Bapat V A,Trivedi P K,Ghosh A,Sane V A,Thumballi G R,Nath P.Ripening of fleshy fruit:molecular insight and the role of ethylene[J].Biotechnology Advances,2010,28(1):94-107.doi:10.1016/j.biotechadv.2009.10.002.
[13] Haji T,Yaegaki H,Yamaguchi M.Changes in ethylene production and flesh firmness of melting,nonmelting and stony hard peaches after harvest[J].Journal of the Japanese Society for Horticultural Science,2001,70:458-459.doi:10.2503/jjshs.70.458.
[14] Goffreda J C.Stony hard gene of peach alters ethylene biosynthesis,respirationand oyher ripening related gharacteristic[J].HortScience,1992,27(6):610-610.
[15] Tsunashima Y,Owino W,Kubo Y,Inaba A,Mathooko F.Regulation of genes encoding ethylene biosynthetic enzymes in peach(Prunus persica L.)fruit by carbon dioxide and 1-methylcyclopropene[J].Postharvest Biology & Technology,2001,21(3):265-281.doi:10.1016/S0925-5214(00)00158-7.
[16] Tatsuki M,Haji T,Yamaguchi M.The involvement of 1-aminocyclopropane-1-carboxylic acid synthase isogene,PpACS1,in peach fruit softening[J].Journal of Experimental Botany,2006,57(6):1281-1289.doi:10.1093/jxb/erj097.
[17] Ohmiya A,Haji T.Promotion of ethylene biosynthesis in peach mesocarp discs by auxin[J].Plant Growth Regulation,2002,36(3):209-214.doi:10.1023/a:1016526517435.
[18] Pan L,Zeng W F,Niu L,Lu Z H,Liu H,Cui G C,Zhu Y Q,Chu J F,Li W P,Fang W C,Cai Z G,Li G H,Wang Z Q.PpYUC11,a strong candidate gene for the stony hard phenotype in peach(Prunus persica L.Batsch),participates in IAA biosynthesis during fruit ripening[J].Journal of Experimental Botany,2015,66(22):7031-7044.doi:10.1093/jxb/erv400.
[19] Marco C,Daniela G,Angelo C,Remo C,Stefano G,Laura R,Alessandro L,Daniele B.Integrative genomics approaches validate PpYUC11-like as candidate gene for the stony hard trait in peach (P.persica L.Batsch)[J].BMC Plant Biology,2018,18(1):88.doi:10.1186/s12870-018-1293-6.
[20] Kenneth J L,Thomas D S.Analysis of relative gene expression data using real-time quantitative PCR and the 2-ΔΔCT,method[J].Methods-A Companion To Methods in Enzymology,2001,25(4):402-408.doi:10.1006/meth.2001.1262.
[21] Mathooko F M,Tsunashima Y,Owino W Z O,Kubo Y,Inaba A.Regulation of genes encoding ethylene biosynthetic enzymes in peach(Prunus persica L.)fruit by carbon dioxide and 1-methylcyclopropene[J].Postharvest Biology and Technology,2001,21(3):265-281.doi:10.1016/S0925-5214(00)00158-7.
[22] 杨勇,马瑞娟,张斌斌,宋志忠,张春华,郭绍雷.不同溶质桃果实的软化与乙烯合成相关基因的差异表达[J].园艺学报,2015,42(10):1869-1878.doi:10.16420/j.issn.0513-353x.2015-0126.Yang Y,Ma R J,Zhang B B,Song Z Z,Zhang C H,Guo S L.Differential expression analysis in fruit softening and ethylene biosynthetic pathways in peaches of different flesh textures[J].Acta Horticulturae Sinica,2015,42(10):1869-1878.
[23] Ghiani A,Baldin F,Morgutti S,Negrini N,Nocito F F,Mignani I,Bassi D,Cocucci M.Changes in the expression of genes involved in ethylene signalling in peach fruit with different flesh texture and softening patterns[J].Acta Horticulturae,2010,884:125-131.doi:10.17660/ActaHortic.2010.884.13.
[24] Tatsuki M,Haji T,Yamaguchi M.The involvement of 1-aminocyclopropane-1-carboxylic acid synthase isogene,PpACS1,in peach fruit softening[J].Journal of Experimental Botany,2006,57(6):1281-1289.doi:10.1093/jxb/erj097.
[25] Liu M C,Pirrello J,Chervin C,Roustan J P,Bouzayen M.Ethylene control of fruit ripening:Revisiting the complex network of transcriptional regulation[J].Plant Physiology,2015,169(4):2380.doi:10.1104/pp.15.01361.
[26] 苏素香,赵彩平,曹丽军,李金金,李芳,韩明玉.两种不同耐贮性桃果实采后乙烯合成和果实软化相关基因表达的差异[J].农业生物技术学报,2015,23(4):450-458.doi:10.3969/j.issn.1674-7968.2015.04.004.Su S X,Zhao C P,Cao L J,Li J J,Li F,Han M Y.The difference of ethylene biosynthesis and expression of softening-related genes between two peach(Prunus persica)cultivars with different storage property after harvest[J].Journal of Agricultural Biotechnology,2015,23(4):450-458.
[27] 曾文芳,丁义峰,潘磊,王小贝,牛良,鲁振华,崔国朝,王志强.桃硬质性状可能源于PpYUC11基因启动子区域CACTA型转座子的插入[J].果树学报,2017,34(10):1239-1248.doi:10.13925/j.cnki.gsxb.20170060.Zeng W F,Ding Y F,Pan L,Wang X B,Niu L,Lu Z H,Cui G C,Wang Z Q.A CACTA transposable element in a PpYUC11 gene promoter is associated with the stony hard phenotype in peach[J].Journal of Fruit Science,2017,34(10):1239-1248.
[28] Tatsuki M,Nakajima N,Fujii H,Shimada T,Nakano M,Hayashi K,Hayama H,Yoshioka H,Nakamura Y.Increased levels of IAA are required for system 2 ethylene synthesis causing fruit softening in peach(Prunus persica L.Batsch)[J].Journal of Experimental Botany,2013,64(4):1049-1059.doi:10.1093/jxb/ers381.