CPPU处理猕猴桃常温贮藏过程中果实糖含量相关基因的表达分析
|
摘要
为了解析采前CPPU处理的猕猴桃果实贮藏性与未处理果实的差异及相关的分子调控机制,以清水为对照,评价了盛花后用20 mg/L CPPU处理果实在常温贮藏下可溶性固形物、可滴定酸及可溶性糖含量的变化,并用转录组分析鉴定出与可溶性糖变化相关的候选基因。结果表明,无论处理还是对照,可溶性糖含量在贮藏过程中呈逐渐上升趋势,但CPPU处理果实的可溶性糖含量始终低于对照果实。转录组分析表明,CPPU处理抑制了淀粉和蔗糖代谢中的基因Achn087691(编码磷酸己糖异构酶)以及果糖和甘露糖代谢中基因Achn203191(编码磷酸丙糖异构酶)的表达,并促进淀粉和蔗糖代谢中基因Achn069851(编码己糖激酶)前期表达,但对基因Achn161931(编码尿苷二磷酸葡萄糖脱氢酶)、基因Achn206141(编码尿苷二磷酸-葡萄糖醛酸酯-4-表异构酶)和基因Achn295291(编码果胶甲酯酶)表现为前期抑制、中期促进、后期抑制。由此推测,20 mg/L CPPU处理明显影响了徐香猕猴桃果实中可溶性糖含量及糖代谢相关基因的表达,致使果实常温贮藏过程中可溶性固形物和可溶性糖含量降低,且提前软化。
In order to study the regulation mechanism in the sugar meta bolismin of CPPU treated kiwifruit during the storage,this study analyzed the differences in total soluble solids( TSS),titratable acid( TA) and soluble sugar conten( SSC) between 20 mg/L CPPU-treated and water-treated kiwifruits after two,four,six,or eight days of storage,and differential gene expression related to these compounds using high-throughput sequencing. The study found that,regardless of the treatment or the control,the soluble sugar content gradually increased during the storage,but the soluble sugar content of the CPPU-treated fruit was always lower than that of the control fruit. CPPU treatment inhibited the expression of gene Achn087691( glucose-6-phosphate isomerase) in starch and sucrose metabolism and Achn203191( triosephosphate isomeras) in fructose and mannose metabolism, and promoted early expression of gene Achn069851( hexokinase) in starch and sucrose metabolism,but showed early inhibition,medium-term promotion,late suppression of gene Achn161931( UDP-glucose 6-dehydrogenase),gene Achn206141( udp-galactose-4-epimerase),and gene Achn295291( pectin methylesterase). It was speculated that 20 mg/L CPPU treatment significantly affected the soluble sugar content and sugar metabolism related gene expression in kiwifruit( Actinidia deliciosa Xuxiang),resulting in a decrease in TSS and SSC during storage at room temperature and accelerated the softening of fruit.
In order to study the regulation mechanism in the sugar meta bolismin of CPPU treated kiwifruit during the storage,this study analyzed the differences in total soluble solids( TSS),titratable acid( TA) and soluble sugar conten( SSC) between 20 mg/L CPPU-treated and water-treated kiwifruits after two,four,six,or eight days of storage,and differential gene expression related to these compounds using high-throughput sequencing. The study found that,regardless of the treatment or the control,the soluble sugar content gradually increased during the storage,but the soluble sugar content of the CPPU-treated fruit was always lower than that of the control fruit. CPPU treatment inhibited the expression of gene Achn087691( glucose-6-phosphate isomerase) in starch and sucrose metabolism and Achn203191( triosephosphate isomeras) in fructose and mannose metabolism, and promoted early expression of gene Achn069851( hexokinase) in starch and sucrose metabolism,but showed early inhibition,medium-term promotion,late suppression of gene Achn161931( UDP-glucose 6-dehydrogenase),gene Achn206141( udp-galactose-4-epimerase),and gene Achn295291( pectin methylesterase). It was speculated that 20 mg/L CPPU treatment significantly affected the soluble sugar content and sugar metabolism related gene expression in kiwifruit( Actinidia deliciosa Xuxiang),resulting in a decrease in TSS and SSC during storage at room temperature and accelerated the softening of fruit.
引文
[1]王玮,何宜恒,李桦,梁春强,饶景萍. CPPU处理对‘华优’猕猴桃品质及耐贮性的影响[J].食品科学,2016,37(6):261-266. doi:10. 7506/spkx1002-6630-201606047.Wang W,He Y H,Li H,Liang C Q,Rao J P. Effect of-(2-Chloropyridin-4-yl)-3-phenylurea(CPPU)treatmenton postharvest fruit quality and storability in‘Huayou’ki-wifruit[J]. Food Science,2016,37(6):261-266.doi:10. 7506/spkx1002-6630-201606047.
[2]王玮,靳蜜静,饶景萍. CPPU处理对采后猕猴桃品质及耐冷性的影响[J].北方园艺,2016(1):11-117. doi:10. 11937/bfyy. 201601029.Wang W,Jin M J,Rao J P. Effect of CPPU treatment onpostharvest fruit quality and chilling resistance in kiwifruit[J]. Northern Horticulture,2016(1):11-117. doi:10. 11937/bfyy. 201601029.
[3]侯勇,马国瑞,夏中梅,曾显斌. CPPU研究进展[J].植物营养与肥料学报,1999,5(2):11-19. doi:10.11674/zwyf. 1999. 02. 003.Hou Y,Ma G R,Xia Z M,Zeng X B. Advances in CPPUresearch[J]. Plant Nutrition and Fertilizer Science,1999,5(2):11-19. doi:10. 11674/zwyf. 1999. 02. 003.
[4] Peppi M C,Fidelibus M W. Effects of forchlorfenuronand abscisic acid on the quality of'Flame Seedless'grapes[J]. Hort Science,2008,43(1):173-176.
[5]赵治兵,王金华,刘永玲,李莹,马立志.基于主成分分析法研究氯吡脲对猕猴桃品质的影响[J].食品工业,2018,39(9):190-193.Zhao Z B,Wang J H,Liu Y L,Li Y,Ma L Z. Explorethe influences of forchlorfenuron on the kiwi fruit qualitybased on principal component analysis method[J]. TheFood Industry,2018,39(9):190-193.
[6]董金磊,郭文川.不同质量浓度氯吡脲对猕猴桃果实理化参数的影响[J].西北农林科技大学学报(自然科学版),2015,43(10):146-151. doi:10. 13207/j.cnki. jnwafu. 2015. 10. 019.Dong J L,Guo W C. Effects of forchlorfenuron concen-tration on physical and chemical properties of kiwifruits[J]. Journal of Northwest A&F University(Natural Sci-ence Edition),2015,43(10):146-151. doi:10.13207/j. cnki. jnwafu. 2015. 10. 019.
[7] Antognozzi E,Battistelli A,Fam I F,et al. Influence ofCPPU on carbohydrate accumulation and metabolism infruits of A ctinidia deliciosa(A. Chev.)[J]. ScientiaHorticulture,1996(95):00852-00857. doi:org/10.1016/0304-4238(95)00852-7.
[8]郭叶,王亚萍,费学谦,刘力.不同浓度CPPU处理对“徐香”猕猴桃贮藏生理和品质的影响[J].食品工业科技,2012,33(20):324-327,354. doi:10. 13386/j. issn1002-0306. 2012. 20. 007.Guo S,Wang Y P,Fei X Q,Liu L. Effect of differentconcentrations of CPPU on physiological and nutritionalquality of"xuxiang"kiwifruit[J]. Science and Technolo-gy of Food Industry,2012,33(20):324-327,354.doi:10. 13386/j. issn1002-0306. 2012. 20. 007.
[9]朱杰丽,杨柳,柴振林.氯吡脲对“徐香”猕猴桃果实品质的影响[J].福建林业科技,2014,41(1):114-165. doi:10. 13428/j. cnki. fjlk. 2014. 01. 024.Zhu J L,Yang L,Chai Z L. Effect of forchlorfenuron onXuxiang actinidia deliciosa fruit quality[J]. Journal ofFujian Forestry Science and Technology,2014,41(1):114-165. doi:10. 13428/j. cnki. fjlk. 2014. 01. 024.
[10] Kim J G,Takami Y,Mizugami T,Beppu K,FukudaT,Kataoka I. CPPU application on size and quality ofhardy kiwifruit[J]. Scientia Horticulturae,2006,110:219-222. doi:10. 1016/j. scienta. 2006. 06. 017.
[11]聂继云,李静,徐国峰,李海飞. NY/T 2637-2014.水果和蔬菜可溶性固形物含量的测定折射仪法[S].北京:中国农业出版社,2015.Nie J Y,Li J,Xu G F,Li H F. NY/T 2637-2014.Determination of soluble solids in fruits and vegetables byRefractometer[S]. Beijing:China Agriculture Press,2015
[12]龚玲娣,徐清渠. GB/T 12456-2008.食品中总酸的测定[S].北京:中国标准出版社,2008.Gong L D,Xu Q Q,GB/T 12456-2008. Determina-tion of total acid in foods[S]. Beijing:Standards Pressof China,2008.
[13] Kim D,Pertea G,Trapnell C,Pimentel H,Kelleyr R,Salzberg S L. Top Hat2:accurate alignment of transcrip-tomes in the presence of insertions,deletions and genefusions[J]. Genome Biology,2013,14(4):R36. doi:10. 1186/gb-2013-14-4-r36.
[14] Huang S,Ding J,Deng D,Tang W,Sun H,Liu D,Zhang L,Niu X,Zhang X,Meng M,Yu J,Liu J,HanY,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,Huang L,SongJ,Xu C,Li J,Ye K,Zhong S,Lu B R,He G,Xiao F,Wang H L,Zheng H,Fei Z,Liu Y. Draft genome of thekiwifruit Actinidia chinensis[J]. Nature Communications,2013,4(2013):2640. doi:10. 1038/ncomms3640.
[15] Conesa A and Gotz S. Blast2GO:acomprehensive suitefor functional analysis in plant genomics[J]. Interna-tional Journal of Plant Genomics,2008,619-632. doi:10. 1155/2008/619832.
[16] Luo J,Guo L L,Huang Y A,Wang C,Qiao C K,PangR L,Li J,Pang T,Wang R P,Xie H Z,Fang J B.Transcriptome analysis reveals the effect of pre-harvestCPPU treatment on the volatile compounds emitted bykiwifruit stored at room temperature[J]. Food ResearchInternational,2017,102:666-673. doi:10. 1016/j. foo-dres. 2017. 09. 051.
[17] Cruz-Castillo J G,Baldicchi A,Frioni T,Marocchi F,Moscatello S,Proietti S,Battistelli A,Famiani F. Pre-anthesis CPPU low dosage application increases ‘hay-ward’kiwifruit weight without affecting the other qualita-tive and nutritional characteristics[J]. Foodchem,2014(158):224-228. doi:10.1016/j. foodchem.2014.01.131.
[18]潘俨,孟新涛,车凤斌,薛素琳,张婷,赵世荣,廖康.库尔勒香梨果实发育成熟的糖代谢和呼吸代谢响应特征[J].中国农业科学,2016,49(17):3391-3412. doi:10. 3864/j. issn. 0578-1752. 2016. 17. 013.Pan Y,Meng X T,Che F B,Xue S L,Zhang T,ZhaoS R,Liao K. Metabolic profiles of sugar metabolism andrespiratory metabolism of korla pear(Pyrus sinkiangensisYu)throughout fruit development and ripening[J]. Sci-entia Agricultura Sinica,2016,49(17):3391-3412.doi:10. 3864/j. issn. 0578-1752. 2016. 17. 013.
[19]秦巧平,张上隆,陈俊伟,吴延军,陈昆松.温州蜜柑果实发育期间果糖激酶与糖积累的关系[J].植物生理与分子生物学学报,2004,30(4):435-440.doi:10. 3321/j. issn:1671-3877. 2004. 04. 011.Qin Q P,Zhang S L,Chen J W,Wu Y J,Chen K S. Therelationship of fructokinase and sugar accumulation duringfruit development in satsuma Mandarin[J]. Journal ofPlant Physiology and Molecular Biology,2004,30(4):435-440. doi:10. 3321/j. issn:1671-3877. 2004. 04. 011.
[20] Van Buggenhout S,Sila D N,Duvetter T,Van Loey A,Hendrickx M. Pectins in processed fruits and vegetables:Part ill-texture engineering[J]. Comprehensive Reviewsin Food Scicnce and Food Safety,2009,8(2):1541-4337. doi:org/10. 1111/j. 1541-4337. 2009. 00072. x.
[21]曾秀丽,张光伦,吕秀兰,龚荣高,罗楠,胡强. 3类生境下脐橙果实细胞壁酶、瓤囊壁超微结构和品质变化的研究[J].果树学报,2006,23(5):670-675. doi:10. 13925/j. cnki. gsxb. 2006. 05. 003.Zeng X L,Zhang G L,Lv X L,Gong R G,Luo N,HuQ. Cell wall enzyme activity,cell ultrastructure and fruitquality of Robertson navel orange grown in three differenthabitats[J]. Journal of Fruit Science,2006,23(5):670-675. doi:10. 13925/j. cnki. gsxb. 2006. 05. 003.
[22] Dong T,Xia R X,Xiao Z Y,Wang P,Song W H.Effect of pre-harvest application of Calcium and Boronon dietary fibre,hydrolases and ultrastructure in`CaraCara'navel orange(Citrus sinensis L. Osbeck)fruit[J]. Scientia Horticulturae,2009,121(3):272-277.doi:10. 1016/j. scienta. 2009. 02. 003.
[23]庞荣,任亚梅,袁春龙,师俊玲,樊明涛.膨大剂处理对六种猕猴桃采收和软熟时品质的影响[J].现代食品科技,2017,33(8):1673-9078. doi:10.13982/j. mfst. 1673-9078. 2017. 8. 034.Pang R,Ren Y M,Yuan C L,Shi J L,Fan M T. Effectsof forchlorfenuron treatment on the quality of six varieties ofkiwifruit during harvest and ripening periods[J]. ModernFood Science and Technology,2017,33(8):1673-9078.doi:10. 13982/j. mfst. 1673-9078. 2017. 8. 034.
[2]王玮,靳蜜静,饶景萍. CPPU处理对采后猕猴桃品质及耐冷性的影响[J].北方园艺,2016(1):11-117. doi:10. 11937/bfyy. 201601029.Wang W,Jin M J,Rao J P. Effect of CPPU treatment onpostharvest fruit quality and chilling resistance in kiwifruit[J]. Northern Horticulture,2016(1):11-117. doi:10. 11937/bfyy. 201601029.
[3]侯勇,马国瑞,夏中梅,曾显斌. CPPU研究进展[J].植物营养与肥料学报,1999,5(2):11-19. doi:10.11674/zwyf. 1999. 02. 003.Hou Y,Ma G R,Xia Z M,Zeng X B. Advances in CPPUresearch[J]. Plant Nutrition and Fertilizer Science,1999,5(2):11-19. doi:10. 11674/zwyf. 1999. 02. 003.
[4] Peppi M C,Fidelibus M W. Effects of forchlorfenuronand abscisic acid on the quality of'Flame Seedless'grapes[J]. Hort Science,2008,43(1):173-176.
[5]赵治兵,王金华,刘永玲,李莹,马立志.基于主成分分析法研究氯吡脲对猕猴桃品质的影响[J].食品工业,2018,39(9):190-193.Zhao Z B,Wang J H,Liu Y L,Li Y,Ma L Z. Explorethe influences of forchlorfenuron on the kiwi fruit qualitybased on principal component analysis method[J]. TheFood Industry,2018,39(9):190-193.
[6]董金磊,郭文川.不同质量浓度氯吡脲对猕猴桃果实理化参数的影响[J].西北农林科技大学学报(自然科学版),2015,43(10):146-151. doi:10. 13207/j.cnki. jnwafu. 2015. 10. 019.Dong J L,Guo W C. Effects of forchlorfenuron concen-tration on physical and chemical properties of kiwifruits[J]. Journal of Northwest A&F University(Natural Sci-ence Edition),2015,43(10):146-151. doi:10.13207/j. cnki. jnwafu. 2015. 10. 019.
[7] Antognozzi E,Battistelli A,Fam I F,et al. Influence ofCPPU on carbohydrate accumulation and metabolism infruits of A ctinidia deliciosa(A. Chev.)[J]. ScientiaHorticulture,1996(95):00852-00857. doi:org/10.1016/0304-4238(95)00852-7.
[8]郭叶,王亚萍,费学谦,刘力.不同浓度CPPU处理对“徐香”猕猴桃贮藏生理和品质的影响[J].食品工业科技,2012,33(20):324-327,354. doi:10. 13386/j. issn1002-0306. 2012. 20. 007.Guo S,Wang Y P,Fei X Q,Liu L. Effect of differentconcentrations of CPPU on physiological and nutritionalquality of"xuxiang"kiwifruit[J]. Science and Technolo-gy of Food Industry,2012,33(20):324-327,354.doi:10. 13386/j. issn1002-0306. 2012. 20. 007.
[9]朱杰丽,杨柳,柴振林.氯吡脲对“徐香”猕猴桃果实品质的影响[J].福建林业科技,2014,41(1):114-165. doi:10. 13428/j. cnki. fjlk. 2014. 01. 024.Zhu J L,Yang L,Chai Z L. Effect of forchlorfenuron onXuxiang actinidia deliciosa fruit quality[J]. Journal ofFujian Forestry Science and Technology,2014,41(1):114-165. doi:10. 13428/j. cnki. fjlk. 2014. 01. 024.
[10] Kim J G,Takami Y,Mizugami T,Beppu K,FukudaT,Kataoka I. CPPU application on size and quality ofhardy kiwifruit[J]. Scientia Horticulturae,2006,110:219-222. doi:10. 1016/j. scienta. 2006. 06. 017.
[11]聂继云,李静,徐国峰,李海飞. NY/T 2637-2014.水果和蔬菜可溶性固形物含量的测定折射仪法[S].北京:中国农业出版社,2015.Nie J Y,Li J,Xu G F,Li H F. NY/T 2637-2014.Determination of soluble solids in fruits and vegetables byRefractometer[S]. Beijing:China Agriculture Press,2015
[12]龚玲娣,徐清渠. GB/T 12456-2008.食品中总酸的测定[S].北京:中国标准出版社,2008.Gong L D,Xu Q Q,GB/T 12456-2008. Determina-tion of total acid in foods[S]. Beijing:Standards Pressof China,2008.
[13] Kim D,Pertea G,Trapnell C,Pimentel H,Kelleyr R,Salzberg S L. Top Hat2:accurate alignment of transcrip-tomes in the presence of insertions,deletions and genefusions[J]. Genome Biology,2013,14(4):R36. doi:10. 1186/gb-2013-14-4-r36.
[14] Huang S,Ding J,Deng D,Tang W,Sun H,Liu D,Zhang L,Niu X,Zhang X,Meng M,Yu J,Liu J,HanY,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,Huang L,SongJ,Xu C,Li J,Ye K,Zhong S,Lu B R,He G,Xiao F,Wang H L,Zheng H,Fei Z,Liu Y. Draft genome of thekiwifruit Actinidia chinensis[J]. Nature Communications,2013,4(2013):2640. doi:10. 1038/ncomms3640.
[15] Conesa A and Gotz S. Blast2GO:acomprehensive suitefor functional analysis in plant genomics[J]. Interna-tional Journal of Plant Genomics,2008,619-632. doi:10. 1155/2008/619832.
[16] Luo J,Guo L L,Huang Y A,Wang C,Qiao C K,PangR L,Li J,Pang T,Wang R P,Xie H Z,Fang J B.Transcriptome analysis reveals the effect of pre-harvestCPPU treatment on the volatile compounds emitted bykiwifruit stored at room temperature[J]. Food ResearchInternational,2017,102:666-673. doi:10. 1016/j. foo-dres. 2017. 09. 051.
[17] Cruz-Castillo J G,Baldicchi A,Frioni T,Marocchi F,Moscatello S,Proietti S,Battistelli A,Famiani F. Pre-anthesis CPPU low dosage application increases ‘hay-ward’kiwifruit weight without affecting the other qualita-tive and nutritional characteristics[J]. Foodchem,2014(158):224-228. doi:10.1016/j. foodchem.2014.01.131.
[18]潘俨,孟新涛,车凤斌,薛素琳,张婷,赵世荣,廖康.库尔勒香梨果实发育成熟的糖代谢和呼吸代谢响应特征[J].中国农业科学,2016,49(17):3391-3412. doi:10. 3864/j. issn. 0578-1752. 2016. 17. 013.Pan Y,Meng X T,Che F B,Xue S L,Zhang T,ZhaoS R,Liao K. Metabolic profiles of sugar metabolism andrespiratory metabolism of korla pear(Pyrus sinkiangensisYu)throughout fruit development and ripening[J]. Sci-entia Agricultura Sinica,2016,49(17):3391-3412.doi:10. 3864/j. issn. 0578-1752. 2016. 17. 013.
[19]秦巧平,张上隆,陈俊伟,吴延军,陈昆松.温州蜜柑果实发育期间果糖激酶与糖积累的关系[J].植物生理与分子生物学学报,2004,30(4):435-440.doi:10. 3321/j. issn:1671-3877. 2004. 04. 011.Qin Q P,Zhang S L,Chen J W,Wu Y J,Chen K S. Therelationship of fructokinase and sugar accumulation duringfruit development in satsuma Mandarin[J]. Journal ofPlant Physiology and Molecular Biology,2004,30(4):435-440. doi:10. 3321/j. issn:1671-3877. 2004. 04. 011.
[20] Van Buggenhout S,Sila D N,Duvetter T,Van Loey A,Hendrickx M. Pectins in processed fruits and vegetables:Part ill-texture engineering[J]. Comprehensive Reviewsin Food Scicnce and Food Safety,2009,8(2):1541-4337. doi:org/10. 1111/j. 1541-4337. 2009. 00072. x.
[21]曾秀丽,张光伦,吕秀兰,龚荣高,罗楠,胡强. 3类生境下脐橙果实细胞壁酶、瓤囊壁超微结构和品质变化的研究[J].果树学报,2006,23(5):670-675. doi:10. 13925/j. cnki. gsxb. 2006. 05. 003.Zeng X L,Zhang G L,Lv X L,Gong R G,Luo N,HuQ. Cell wall enzyme activity,cell ultrastructure and fruitquality of Robertson navel orange grown in three differenthabitats[J]. Journal of Fruit Science,2006,23(5):670-675. doi:10. 13925/j. cnki. gsxb. 2006. 05. 003.
[22] Dong T,Xia R X,Xiao Z Y,Wang P,Song W H.Effect of pre-harvest application of Calcium and Boronon dietary fibre,hydrolases and ultrastructure in`CaraCara'navel orange(Citrus sinensis L. Osbeck)fruit[J]. Scientia Horticulturae,2009,121(3):272-277.doi:10. 1016/j. scienta. 2009. 02. 003.
[23]庞荣,任亚梅,袁春龙,师俊玲,樊明涛.膨大剂处理对六种猕猴桃采收和软熟时品质的影响[J].现代食品科技,2017,33(8):1673-9078. doi:10.13982/j. mfst. 1673-9078. 2017. 8. 034.Pang R,Ren Y M,Yuan C L,Shi J L,Fan M T. Effectsof forchlorfenuron treatment on the quality of six varieties ofkiwifruit during harvest and ripening periods[J]. ModernFood Science and Technology,2017,33(8):1673-9078.doi:10. 13982/j. mfst. 1673-9078. 2017. 8. 034.