Adβgal-1和Adβgal-2克隆及其在猕猴桃果实软化中的作用
|
摘要
【目的】β-半乳糖苷酶是一类参与细胞壁降解的糖苷酶,在植物的生长发育和果实成熟软化过程中发挥重要作用。通过对猕猴桃2个β-半乳糖苷酶基因的鉴定及其表达模式研究,明确它们在猕猴桃果实软化中的作用,丰富猕猴桃的后熟软化机理研究。【方法】以‘米良1号’猕猴桃为试材,采用RT-PCR法扩增果实的β-半乳糖苷酶基因,利用在线数据库分析其编码蛋白的特性、功能结构域、系统进化关系、基因结构和调控miRNA,应用qPCR研究其在不同组织部位、果实的不同软化时期、不同贮藏温度和ABA处理后的表达特征,并结合酶活性变化,阐释这2个β-半乳糖苷酶基因在猕猴桃果实软化中的作用。【结果】克隆得到2个猕猴桃β-半乳糖苷酶基因(Adβgal-1和Adβgal-2),登录号分别为MH319788和MH319789。Adβgal-1的开放阅读框为2 280 bp,编码759个氨基酸;Adβgal-2的开放阅读框为2 025 bp,编码674个氨基酸。结构域分析显示,Adβgal-1和Adβgal-2均含有植物糖苷水解酶家族35的功能结构域(Glyco_hydro_35)。基因结构分析表明,Adβgal-1由18个外显子和17个内含子组成,而Adβgal-2由17个外显子和16个内含子组成。进化树分析显示它们位于两个不同的进化分支中,且序列差异较大,可能源自不同的基因祖先。qPCR分析结果表明,Adβgal-1和Adβgal-2在猕猴桃的各组织部位(根、茎、叶、花、幼果、成熟果)均有表达,但表达水平不同;它们均在果实软化初期上调表达,随着果实硬度的下降,表达量持续增加;在25℃贮藏过程中,它们均在3 d时上调表达,随着时间的推移,表达量增加;而在4℃贮藏过程中,Adβgal-1的表达受到抑制,Adβgal-2的表达呈波浪式;ABA处理诱导Adβgal-1和Adβgal-2的表达。酶活性测定结果显示,β-半乳糖苷酶活性在果实软化初期略有降低,随着果实硬度的下降逐渐升高。【结论】Adβgal-1和Adβgal-2均参与猕猴桃果实的软化进程。不同贮藏温度和ABA处理对猕猴桃果实软化的影响可能是通过改变β-半乳糖苷酶基因的表达而实现的。
【Objective】Beta-galactosidase is one type of glycosidases involving in cell wall degradation, which plays an important role in plant growth and fruit ripening and softening. In this paper, the identification and expression analysis of two β-galactosidase genes in kiwifruit, as well as their functions during fruit softening, were carried out to provide new insights for the mechanism of kiwifruit softening.【Method】Actinidia chinensis var. deliciosa ‘Miliang-1' was used as material to amplify the β-galactosidase genes by RT-PCR, then their characteristics, functional domains, phylogenetic relationships, gene structures and miRNAs were analyzed by using online databases. qRCR was applied to analyze their expression patterns in different tissues, different stages of softening, different storage temperatures, and ABA treatment. Enzymes activities were also analyzed to clarify the function of these two β-galactosidase genes during fruit softening.【Results】Two β-galactosidase genes(Adβgal-1 and Adβgal-2) were obtained from kiwifruit. Their accession numbers were MH319788 and MH319789, respectively. Adβgal-1 had an open reading frame(ORF) of 2 280 bp and encoded 759 amino acids, while the ORF of Adβgal-2 was 2 025 bp encoding 674 amino acids. Protein domain analysis showed that both Adβgal-1 and Adβgal-2 harbored a functional domain(Glyco_hydro_35) of plant glycoside hydrolase family 35. Gene organization analysis revealed that Adβgal-1 had 18 exons and 17 introns, while Adβgal-2 harbored 17 exons and 16 introns. Moreover, Adβgal-1 and Adβgal-2 were clustered in different evolutionary branching in the phylogenetic tree, which was due to the great difference in their sequences. This implied that Adβgal-1 and Adβgal-2 might originate from different ancestral genes. qPCR analysis showed that Adβgal-1 and Adβgal-2 were expressed in all tested tissues(root, stem, leaf, flower, young fruit and ripe fruit) although their expression levels were different. The expression levels of both Adβgal-1 and Adβgal-2 were increased at the start stage of fruit softening, and the amount of gene expression continued to increase as the firmness of fruit decreased. When stored at 25℃, both the Adβgal-1 and Adβgal-2 were upregulated at 3 d, and the amount of gene expression continued to increase as time over. When stored at 4℃, the expression of Adβgal-1 was inhabited, while that of Adβgal-2 was like wavy. ABA induced the expression of Adβgal-1 and Adβgal-2.【Conclusion】Both Adβgal-1 and Adβgal-2 participated in the process of fruit softening of kiwifruit. The effect of storage temperatures and ABA on fruit softening might be achieved by changing the expression of β-galactosidase genes.
【Objective】Beta-galactosidase is one type of glycosidases involving in cell wall degradation, which plays an important role in plant growth and fruit ripening and softening. In this paper, the identification and expression analysis of two β-galactosidase genes in kiwifruit, as well as their functions during fruit softening, were carried out to provide new insights for the mechanism of kiwifruit softening.【Method】Actinidia chinensis var. deliciosa ‘Miliang-1' was used as material to amplify the β-galactosidase genes by RT-PCR, then their characteristics, functional domains, phylogenetic relationships, gene structures and miRNAs were analyzed by using online databases. qRCR was applied to analyze their expression patterns in different tissues, different stages of softening, different storage temperatures, and ABA treatment. Enzymes activities were also analyzed to clarify the function of these two β-galactosidase genes during fruit softening.【Results】Two β-galactosidase genes(Adβgal-1 and Adβgal-2) were obtained from kiwifruit. Their accession numbers were MH319788 and MH319789, respectively. Adβgal-1 had an open reading frame(ORF) of 2 280 bp and encoded 759 amino acids, while the ORF of Adβgal-2 was 2 025 bp encoding 674 amino acids. Protein domain analysis showed that both Adβgal-1 and Adβgal-2 harbored a functional domain(Glyco_hydro_35) of plant glycoside hydrolase family 35. Gene organization analysis revealed that Adβgal-1 had 18 exons and 17 introns, while Adβgal-2 harbored 17 exons and 16 introns. Moreover, Adβgal-1 and Adβgal-2 were clustered in different evolutionary branching in the phylogenetic tree, which was due to the great difference in their sequences. This implied that Adβgal-1 and Adβgal-2 might originate from different ancestral genes. qPCR analysis showed that Adβgal-1 and Adβgal-2 were expressed in all tested tissues(root, stem, leaf, flower, young fruit and ripe fruit) although their expression levels were different. The expression levels of both Adβgal-1 and Adβgal-2 were increased at the start stage of fruit softening, and the amount of gene expression continued to increase as the firmness of fruit decreased. When stored at 25℃, both the Adβgal-1 and Adβgal-2 were upregulated at 3 d, and the amount of gene expression continued to increase as time over. When stored at 4℃, the expression of Adβgal-1 was inhabited, while that of Adβgal-2 was like wavy. ABA induced the expression of Adβgal-1 and Adβgal-2.【Conclusion】Both Adβgal-1 and Adβgal-2 participated in the process of fruit softening of kiwifruit. The effect of storage temperatures and ABA on fruit softening might be achieved by changing the expression of β-galactosidase genes.
引文
[1]MCATEE P A,RICHARDSON A C,NIEUWENHUIZEN N J,GUNASEELAN K,HOONG L,CHEN X,ATKINSON R G,BURDON J N,DAVID K M,SCHAFFER R J.The hybrid non-ethylene and ethylene ripening response in kiwifruit(Actinidia chinensis)is associated with differential regulation of MADS-box transcription factors.BMC Plant Biology,2015,15(1):304.
[2]黄文俊,钟彩虹.猕猴桃果实采后生理研究进展.植物科学学报,2017,35(4):622-630.HUANG W J,ZHONG C H.Research advances in the postharvest physiology of kiwifruit.Plant Science Journal,2017,35(4):622-630.(in Chinese)
[3]李东,谭书明,邓毅,张程榕,蒋想.猕猴桃采前处理保鲜技术研究进展.食品工业,2015,36(8):251-255.LI D,TAN S M,DENG Y,ZHANG C R,JIANG X.Advances in preharvest preservation methods for kiwifruit.Food Industry,2015,36(8):251-255.(in Chinese)
[4]朱明月,沈文涛,周鹏.果实成熟软化机理研究进展.分子植物育种,2005,3(3):421-426.ZHU M Y,SHEN W T,ZHOU P.Research advances on mechanism of fruit ripening and softening.Molecular Plant Breeding,2005,3(3):421-426.(in Chinese)
[5]田爱梅,刘金龙,曹家树.植物β-半乳糖苷酶.中国细胞生物学学报,2014,36(5):703-707.TIAN A M,LIU J L,CAO J S.Beta galactosidase in plants.Chinese Journal of Cell Biology,2014,36(5):703-707.(in Chinese)
[6]陆胜民,席玙芳,张耀洲.梅果采后软化与细胞壁组分及其降解酶活性的变化.中国农业科学,2003,36(5):595-598.LU S M,XI Y F,ZHANG Y Z.Softening of green mume flesh and changes of cell wall components and activities of their degrading enzymes during the postharvest period.Scientia Agricultura Sinica,2003,36(5):595-598.(in Chinese)
[7]DAWSON D M,MELTON L D,WATKINS C B.Cell wall changes in nectarines(Prunus persica):Solubilization and depolymerization of pectic and neutral polymers during ripening and in mealy fruit.Plant Physiology,1992,100:1203-1210.
[8]LIDSTER P D,MCRAE K B,DICK A J.Effects of a vacuum infusion of a partially purified beta-galactosidase inhibitor on apple quality.HortScience,1985,20:80-82.
[9]庄军平,苏菁,陈维信.香蕉果实β-半乳糖苷酶基因c DNA克隆及序列分析.西北植物学报,2006,26(1):18-22.ZHUANG J P,SUN J,CHEN W X.cDNA cloning and sequencing ofβ-galactosidase genes in banana(Musa spp.)fruit.Acta Botanica Boreali-occidentalia Sinica,2006,26(1):18-22.(in Chinese)
[10]周厚成,李刚,赵霞,郭蔼光.草莓β-半乳糖苷酶基因FaTβgal的克隆与表达分析.西北植物学报,2015,35(12):2385-2390.ZHOU H C,LI G,ZHAO X,GUO A G.Cloning and expression analysis of Fa Tβgal ofβ-galactosidase family from Fragaria ananassa fruit.Acta Botanica Boreali-occidentalia Sinica,2015,35(12):2385-2390.(in Chinese)
[11]吕燕荣,任小林,周会玲.枣果实β-半乳糖苷酶基因的克隆及表达分析.西北植物学报,2011,31(7):1318-1325.LüY R,REN X L,ZHOU H L.Cloning and expressing analysis ofβ-galactosidase gene in jujube fruit.Acta Botanica Boreali-occidentalia Sinica,2011,31(7):1318-1325.(in Chinese)
[12]高山,陈桂信,许端祥,林碧英,林义章,潘东明.苦瓜果实β-半乳糖苷酶基因的克隆、表达及亚细胞定位.植物遗传资源学报,2013,14(6):1124-1129.GAO S,CHEN G X,XU D X,LIN B Y,LIN Y Z,PAN D M.Cloing,expression and subcellular localization ofβ-galactosidase gene from Momordica charantia L.fruit.Journal of Plant Genetic Resources,2013,14(6):1124-1129.(in Chinese)
[13]卞伟华,江昌俊,王朝霞.桃叶片β-半乳糖苷酶基因全长c DNA克隆及原核表达.园艺学报,2006,33(4):721-725.BIAN W H,JIANG C J,WANG Z C.Cloning and expression ofβ-galactosidase gene cDNA in peach.Acta Horticulturae Sinica,2006,33(4):721-725.(in Chinese)
[14]ZHANG H M,LIU J Y.Molecular cloning and characterization of aβ-galactosidase gene expressed preferentially in cotton fibers.Journal of Integrative Plant Biology,2005,47(2):223-232.
[15]何玮毅,陈晓静,申艳红,卢秉国.番木瓜β-Gal基因的克隆分析与植物表达载体构建.热带作物学报,2010,31(2):217-223.HE W Y,CHEN X J,SHEN Y H,LU B G.Isolation of a papayaβ-Gal gene and construction of plant expresson vector.Chinese Journal of Tropical Crops,2010,31(2):217-223.(in Chinese)
[16]BAN Q,HAN Y,MENG K,HOU Y,HE Y,RAO J.Characterization ofβ-galactosidase genes involved in persimmon growth and fruit ripening and in response to propylene and 1-methylcyclopropene.Journal of Plant Growth Regulation,2016,35(4):1025-1035.
[17]TATEISHI A,NAGASHIMA K.Differential expression of members of the beta-galactosidase gene family during Japanese pear(Pyrus pyrifolia L.)fruit growth and on-tree ripening.Journal of the American Society for Horticultural Science,2005,130(6):819-829.
[18]PANIAGUA C,BLANCO-PORTALES R,BARCELO-MUNOZ M,CARCIA-GAGO J A,WALDRON K W,QUESADA M A,MUNOZ-BLANCO J,MERCADO J A.Antisense down-regulation of the strawberryβ-galactosidase gene FaβGal4 increases cell wall galactose levels and reduces fruit softening.Journal of Experimental Botany,2016,67(3):619-631.
[19]ROSS G S,REDGWELL R J,MACRAE E A.Kiwifruitβ-galactosidase:Isolation and activity against specific fruit cell-wall polysaccharides.Planta,1993,189(4):499-506.
[20]陈昆松,张上隆,Ross Gavin S.β-半乳糖苷酶基因在猕猴桃果实成熟过程的表达.植物生理学报,2000,26(2):117-122.CHEN K S,ZHANG S L,Ross G S.Expression pattern ofβ-galactosidase gene in ripening Actinidia chinensis fruit.Acta Phytophysologica Sinica,2000,26(2):117-122.(in Chinese)
[21]HUANG S X,DING J,DENG D,TANG W,SUN H,LIU D,ZHANGL,NIU X,ZHANG X,MENG M,YU J,LIU J,HAN Y,SHI W,ZHANG D,CAO S,WEI Z,CUI Y,XIA Y,ZENG H,BAO K,LIN L,MIN Y,ZHANG H,MIAO M,TANG X,ZHU Y,SUI Y,LI G,SUN H,YUE J,SUN J,LIU F,ZHOU L,LEI L,ZHENG X,LIU M,HUANGL,SONG J,XU C,LI J,YE K,ZHONG S,LU B,HE G,XIAO F,WANG H,ZHENG H,FEI Z,LIU Y.Draft genome of the kiwifruit Actinidia chinensis.Nature communications,2013,4.doi:10.1038/ncomms3640.
[22]张慧琴,鸣谢,肖金平,周利秋,宋根华.猕猴桃实时荧光定量PCR分析中内参基因的筛选.浙江农业学报,2015,27(4):567-573.ZHANG H Q,XIE M,XIAO J P,ZHOU L Q,SONG G H.Screening of reference genes for real-time quantitative PCR in kiwifruit.Acta Agruiculturae Zhenjiangensis,2015,27(4):567-573.(in Chinese)
[23]DAI X,ZHAO P X.psRNATarget:A plant small RNA target analysis server.Nucleic Acids Research,2011,39(suppl 2):W155-W159.
[24]LIU J H,GAO M H,LV M L,CAO J S.Structure,evolution,and expression of theβ-galactosidase gene family in Brassica campestris ssp.chinensis.Plant Molecular Biology Reporter,2013,31(6):1249-1260.
[25]TATEISHI A,SHIBA H,OGIHARA J,ISOBE K,NOMURA K,WATANABE K,INOUE H.Differential expression and ethylene regulation ofβ-galactosidase genes and isozymes isolated from avocado(Persea americana Mill.)fruit.Postharvest Biology and Technology,2007,45(1):56-65.
[26]MONEO-SANCHEZ M,IZQUIERDO L,MARTIN I,LABRADORE,DOPICO B.Subcellular location of Arabidopsis thaliana subfamily a1β-galactosidases and developmental regulation of transcript levels of their coding genes.Plant Physiology and Biochemistry,2016,109:137-145.
[27]O'Donoghue E M.,SOMERFIELD S D,DEROLES S C,SUTHERLANDP W,HALLETT I C,ERRIDGE Z A,BRUMMELL D A,HUNTER DA.Simultaneous knock-down of sixβ-galactosidase genes in petunia petals prevents loss of pectic galactan but decreases petal strength.Plant Physiology and Biochemistry,2017,13:208-221.
[28]张美芳,何玲,张美丽,郭宇欢.猕猴桃鲜果贮藏保鲜研究进展.食品科学,2014,35(11):343-347.ZHANG M F,HE L,ZHANG M L,GUO Y H.Advances in preservation method for kiwifruit.Food Science,2014,35(11):343-347.(in Chinese)
[29]陈义挺,赖瑞联,程春振,冯新,屈蒙蒙,刘萍元,陈文光,吴如健.猕猴桃果实软化过程中AcPG基因的克隆与表达.农业生物技术学报,2017,25(2):205-213.CHEN Y T,LAI R L,CHENG C Z,FENG X,QU M M,LIU P Y,CHEN W G,WU R J.Cloning and expression of polygalacturonase gene(Ac PG)during fruit softening of kiwifruit(Actinidia chinensis).Journal of Agricultural Biotechnology,2017,25(2):205-213.(in Chinese)
[30]徐秋红.中国李果实软化相关基因的克隆及表达分析[D].南京:南京农业大学,2009.XU Q H.Isolation and expression of genes related to Chinese plum fruit softening[D].Nanjing:Nanjing Agruicultural University,2009.
[31]陈昆松,李方,张上隆.猕猴桃果实成熟进程中木葡聚糖内糖基转移酶m RNA水平的变化.植物学报,1999,41(11):1231-1234.CHEN K S,LI F,ZHANG S L.The expression pattern of xyloglucan endotransglycosylase gene in fruit ripeing of Actinidia chinensis.Acta Botanica Sinica,1999,41(11):1231-1234.(in Chinese)
[32]陈昆松,李方,张上隆,Ross Gavin S.ABA和IAA对猕猴桃果实成熟进程的调控.园艺学报,1999,26(2):81-86.CHEN K S,LI F,ZHANG S L,ROSS G S.Role of abscisic acid and indole-3-acetic acid in kiwifruit ripening.Acta Horticulturae Sinica,1999,26(2):81-86.(in Chinese)
[33]陈昆松,张上隆,吕均良,陈青俊.脱落酸、吲哚乙酸和乙烯在猕猴桃果实后熟软化进程中的变化.中国农业科学,1997,30(2):54-57.CHEN K S,ZHANG S L,LüJ L,CHEN Q J.Variation of abscisic acid,indole-3-acetic acid and ethylene in kiwifruit during fruit ripening.Scientia Agricultura Sinica,1997,30(2):54-57.(in Chinese)
[2]黄文俊,钟彩虹.猕猴桃果实采后生理研究进展.植物科学学报,2017,35(4):622-630.HUANG W J,ZHONG C H.Research advances in the postharvest physiology of kiwifruit.Plant Science Journal,2017,35(4):622-630.(in Chinese)
[3]李东,谭书明,邓毅,张程榕,蒋想.猕猴桃采前处理保鲜技术研究进展.食品工业,2015,36(8):251-255.LI D,TAN S M,DENG Y,ZHANG C R,JIANG X.Advances in preharvest preservation methods for kiwifruit.Food Industry,2015,36(8):251-255.(in Chinese)
[4]朱明月,沈文涛,周鹏.果实成熟软化机理研究进展.分子植物育种,2005,3(3):421-426.ZHU M Y,SHEN W T,ZHOU P.Research advances on mechanism of fruit ripening and softening.Molecular Plant Breeding,2005,3(3):421-426.(in Chinese)
[5]田爱梅,刘金龙,曹家树.植物β-半乳糖苷酶.中国细胞生物学学报,2014,36(5):703-707.TIAN A M,LIU J L,CAO J S.Beta galactosidase in plants.Chinese Journal of Cell Biology,2014,36(5):703-707.(in Chinese)
[6]陆胜民,席玙芳,张耀洲.梅果采后软化与细胞壁组分及其降解酶活性的变化.中国农业科学,2003,36(5):595-598.LU S M,XI Y F,ZHANG Y Z.Softening of green mume flesh and changes of cell wall components and activities of their degrading enzymes during the postharvest period.Scientia Agricultura Sinica,2003,36(5):595-598.(in Chinese)
[7]DAWSON D M,MELTON L D,WATKINS C B.Cell wall changes in nectarines(Prunus persica):Solubilization and depolymerization of pectic and neutral polymers during ripening and in mealy fruit.Plant Physiology,1992,100:1203-1210.
[8]LIDSTER P D,MCRAE K B,DICK A J.Effects of a vacuum infusion of a partially purified beta-galactosidase inhibitor on apple quality.HortScience,1985,20:80-82.
[9]庄军平,苏菁,陈维信.香蕉果实β-半乳糖苷酶基因c DNA克隆及序列分析.西北植物学报,2006,26(1):18-22.ZHUANG J P,SUN J,CHEN W X.cDNA cloning and sequencing ofβ-galactosidase genes in banana(Musa spp.)fruit.Acta Botanica Boreali-occidentalia Sinica,2006,26(1):18-22.(in Chinese)
[10]周厚成,李刚,赵霞,郭蔼光.草莓β-半乳糖苷酶基因FaTβgal的克隆与表达分析.西北植物学报,2015,35(12):2385-2390.ZHOU H C,LI G,ZHAO X,GUO A G.Cloning and expression analysis of Fa Tβgal ofβ-galactosidase family from Fragaria ananassa fruit.Acta Botanica Boreali-occidentalia Sinica,2015,35(12):2385-2390.(in Chinese)
[11]吕燕荣,任小林,周会玲.枣果实β-半乳糖苷酶基因的克隆及表达分析.西北植物学报,2011,31(7):1318-1325.LüY R,REN X L,ZHOU H L.Cloning and expressing analysis ofβ-galactosidase gene in jujube fruit.Acta Botanica Boreali-occidentalia Sinica,2011,31(7):1318-1325.(in Chinese)
[12]高山,陈桂信,许端祥,林碧英,林义章,潘东明.苦瓜果实β-半乳糖苷酶基因的克隆、表达及亚细胞定位.植物遗传资源学报,2013,14(6):1124-1129.GAO S,CHEN G X,XU D X,LIN B Y,LIN Y Z,PAN D M.Cloing,expression and subcellular localization ofβ-galactosidase gene from Momordica charantia L.fruit.Journal of Plant Genetic Resources,2013,14(6):1124-1129.(in Chinese)
[13]卞伟华,江昌俊,王朝霞.桃叶片β-半乳糖苷酶基因全长c DNA克隆及原核表达.园艺学报,2006,33(4):721-725.BIAN W H,JIANG C J,WANG Z C.Cloning and expression ofβ-galactosidase gene cDNA in peach.Acta Horticulturae Sinica,2006,33(4):721-725.(in Chinese)
[14]ZHANG H M,LIU J Y.Molecular cloning and characterization of aβ-galactosidase gene expressed preferentially in cotton fibers.Journal of Integrative Plant Biology,2005,47(2):223-232.
[15]何玮毅,陈晓静,申艳红,卢秉国.番木瓜β-Gal基因的克隆分析与植物表达载体构建.热带作物学报,2010,31(2):217-223.HE W Y,CHEN X J,SHEN Y H,LU B G.Isolation of a papayaβ-Gal gene and construction of plant expresson vector.Chinese Journal of Tropical Crops,2010,31(2):217-223.(in Chinese)
[16]BAN Q,HAN Y,MENG K,HOU Y,HE Y,RAO J.Characterization ofβ-galactosidase genes involved in persimmon growth and fruit ripening and in response to propylene and 1-methylcyclopropene.Journal of Plant Growth Regulation,2016,35(4):1025-1035.
[17]TATEISHI A,NAGASHIMA K.Differential expression of members of the beta-galactosidase gene family during Japanese pear(Pyrus pyrifolia L.)fruit growth and on-tree ripening.Journal of the American Society for Horticultural Science,2005,130(6):819-829.
[18]PANIAGUA C,BLANCO-PORTALES R,BARCELO-MUNOZ M,CARCIA-GAGO J A,WALDRON K W,QUESADA M A,MUNOZ-BLANCO J,MERCADO J A.Antisense down-regulation of the strawberryβ-galactosidase gene FaβGal4 increases cell wall galactose levels and reduces fruit softening.Journal of Experimental Botany,2016,67(3):619-631.
[19]ROSS G S,REDGWELL R J,MACRAE E A.Kiwifruitβ-galactosidase:Isolation and activity against specific fruit cell-wall polysaccharides.Planta,1993,189(4):499-506.
[20]陈昆松,张上隆,Ross Gavin S.β-半乳糖苷酶基因在猕猴桃果实成熟过程的表达.植物生理学报,2000,26(2):117-122.CHEN K S,ZHANG S L,Ross G S.Expression pattern ofβ-galactosidase gene in ripening Actinidia chinensis fruit.Acta Phytophysologica Sinica,2000,26(2):117-122.(in Chinese)
[21]HUANG S X,DING J,DENG D,TANG W,SUN H,LIU D,ZHANGL,NIU X,ZHANG X,MENG M,YU J,LIU J,HAN Y,SHI W,ZHANG D,CAO S,WEI Z,CUI Y,XIA Y,ZENG H,BAO K,LIN L,MIN Y,ZHANG H,MIAO M,TANG X,ZHU Y,SUI Y,LI G,SUN H,YUE J,SUN J,LIU F,ZHOU L,LEI L,ZHENG X,LIU M,HUANGL,SONG J,XU C,LI J,YE K,ZHONG S,LU B,HE G,XIAO F,WANG H,ZHENG H,FEI Z,LIU Y.Draft genome of the kiwifruit Actinidia chinensis.Nature communications,2013,4.doi:10.1038/ncomms3640.
[22]张慧琴,鸣谢,肖金平,周利秋,宋根华.猕猴桃实时荧光定量PCR分析中内参基因的筛选.浙江农业学报,2015,27(4):567-573.ZHANG H Q,XIE M,XIAO J P,ZHOU L Q,SONG G H.Screening of reference genes for real-time quantitative PCR in kiwifruit.Acta Agruiculturae Zhenjiangensis,2015,27(4):567-573.(in Chinese)
[23]DAI X,ZHAO P X.psRNATarget:A plant small RNA target analysis server.Nucleic Acids Research,2011,39(suppl 2):W155-W159.
[24]LIU J H,GAO M H,LV M L,CAO J S.Structure,evolution,and expression of theβ-galactosidase gene family in Brassica campestris ssp.chinensis.Plant Molecular Biology Reporter,2013,31(6):1249-1260.
[25]TATEISHI A,SHIBA H,OGIHARA J,ISOBE K,NOMURA K,WATANABE K,INOUE H.Differential expression and ethylene regulation ofβ-galactosidase genes and isozymes isolated from avocado(Persea americana Mill.)fruit.Postharvest Biology and Technology,2007,45(1):56-65.
[26]MONEO-SANCHEZ M,IZQUIERDO L,MARTIN I,LABRADORE,DOPICO B.Subcellular location of Arabidopsis thaliana subfamily a1β-galactosidases and developmental regulation of transcript levels of their coding genes.Plant Physiology and Biochemistry,2016,109:137-145.
[27]O'Donoghue E M.,SOMERFIELD S D,DEROLES S C,SUTHERLANDP W,HALLETT I C,ERRIDGE Z A,BRUMMELL D A,HUNTER DA.Simultaneous knock-down of sixβ-galactosidase genes in petunia petals prevents loss of pectic galactan but decreases petal strength.Plant Physiology and Biochemistry,2017,13:208-221.
[28]张美芳,何玲,张美丽,郭宇欢.猕猴桃鲜果贮藏保鲜研究进展.食品科学,2014,35(11):343-347.ZHANG M F,HE L,ZHANG M L,GUO Y H.Advances in preservation method for kiwifruit.Food Science,2014,35(11):343-347.(in Chinese)
[29]陈义挺,赖瑞联,程春振,冯新,屈蒙蒙,刘萍元,陈文光,吴如健.猕猴桃果实软化过程中AcPG基因的克隆与表达.农业生物技术学报,2017,25(2):205-213.CHEN Y T,LAI R L,CHENG C Z,FENG X,QU M M,LIU P Y,CHEN W G,WU R J.Cloning and expression of polygalacturonase gene(Ac PG)during fruit softening of kiwifruit(Actinidia chinensis).Journal of Agricultural Biotechnology,2017,25(2):205-213.(in Chinese)
[30]徐秋红.中国李果实软化相关基因的克隆及表达分析[D].南京:南京农业大学,2009.XU Q H.Isolation and expression of genes related to Chinese plum fruit softening[D].Nanjing:Nanjing Agruicultural University,2009.
[31]陈昆松,李方,张上隆.猕猴桃果实成熟进程中木葡聚糖内糖基转移酶m RNA水平的变化.植物学报,1999,41(11):1231-1234.CHEN K S,LI F,ZHANG S L.The expression pattern of xyloglucan endotransglycosylase gene in fruit ripeing of Actinidia chinensis.Acta Botanica Sinica,1999,41(11):1231-1234.(in Chinese)
[32]陈昆松,李方,张上隆,Ross Gavin S.ABA和IAA对猕猴桃果实成熟进程的调控.园艺学报,1999,26(2):81-86.CHEN K S,LI F,ZHANG S L,ROSS G S.Role of abscisic acid and indole-3-acetic acid in kiwifruit ripening.Acta Horticulturae Sinica,1999,26(2):81-86.(in Chinese)
[33]陈昆松,张上隆,吕均良,陈青俊.脱落酸、吲哚乙酸和乙烯在猕猴桃果实后熟软化进程中的变化.中国农业科学,1997,30(2):54-57.CHEN K S,ZHANG S L,LüJ L,CHEN Q J.Variation of abscisic acid,indole-3-acetic acid and ethylene in kiwifruit during fruit ripening.Scientia Agricultura Sinica,1997,30(2):54-57.(in Chinese)