籽用和肉用印度南瓜果实发育过程中淀粉合成途径相关基因的表达分析
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摘要
以籽用印度南瓜自交系98-2及肉用印度南瓜自交系312-1为试材,对印度南瓜果实淀粉含量及淀粉代谢相关基因的表达模式进行研究。结果表明:312-1和98-2果实中的总淀粉和支链淀粉积累模式存在明显差异,312-1中总淀粉和支链淀粉积累模式相似,都是随着果实发育逐渐积累,在授粉后40 d时达到高峰,随后略有下降,而98-2中总淀粉和支链淀粉含量一直维持在果实发育初期(授粉后5 d)水平,没有显著积累的过程;在果实发育的各个时期312-1中总淀粉和支链淀粉含量均极显著高于98-2。2份材料中直链淀粉的积累模式大致相同,都维持在果实发育初期的积累水平,但在果实发育的各个时期312-1中直链淀粉含量均显著或极显著高于98-2。在淀粉组成上,2份材料均是以支链淀粉为主。淀粉合成途径关键酶基因除ISAⅢ、SSSⅡ和SBEⅡ基因外,GPT、AATP、PGM、AGPase L、GBSSⅠ、SSSⅢ和SBEⅠ基因在2份材料果实发育过程中的表达趋势相似,其表达量在2份材料中存在较大差异,结合2份材料果实中直链淀粉和支链淀粉含量,推测GPT、AATP、AGPase L、GBSSⅠ、SSSⅡ、SSSⅢ、SBEⅠ和SBEⅡ基因是调控印度南瓜果实中淀粉积累的关键酶基因。
Taking seed-used pumpkin inbred line ‘98-2' and meat-used pumpkin inbred line ‘312-1' as test materials,this paper studied on the starch content of pumpkin fruit and expression pattern of starch metabolism related genes.The results indicated that there existed significant differences in accumulation patterns of total starch and amylopectin of ‘312-1' and ‘98-2' fruits.The accumulation patterns of total starch and amylopectin in ‘312-1' were similar.They were all accumulated gradually with fruit development,and the peak value appeared in 40 days after pollination,then decreased slightly.While,the contents of total starch and amylopectin in ‘98-2' maintained the same level at the early stage of fruit development(5 days after pollination)without significant accumulation process.The contents of total starch and amylopectin in ‘312-1' were significantly higher than that in ‘98-2' during each fruit development stage.The amylose accumulation pattern in the 2 materials was similar,all maintaining at accumulated level at the early fruit development stage.However,the amylose contents in ‘312-1' at different fruit development stages were significantly or extremely significantly higher than that of ‘98-2'.In starch composition,amylopectin was dominant in the 2 materials.In addition to ISA Ⅲ,SSSⅡ and SBEⅡ genes,the GPT,AATP,PGM,AGPaseL,GBSSⅠ,SSS Ⅲ and SBEⅠ genes had similar expression trends during fruit development process of these 2 materials,and their expression quantities had large differences.Combining the amylopectin and amylopectin contents in fruits of these 2 materials,the paper speculated that GPT,AATP,AGPaseL,GBSSⅠ,SSSⅡ,SSS Ⅲ,SBEⅠ and SBEⅡ genes were key genes regulating starch accumulation in pumpkin fruit.
Taking seed-used pumpkin inbred line ‘98-2' and meat-used pumpkin inbred line ‘312-1' as test materials,this paper studied on the starch content of pumpkin fruit and expression pattern of starch metabolism related genes.The results indicated that there existed significant differences in accumulation patterns of total starch and amylopectin of ‘312-1' and ‘98-2' fruits.The accumulation patterns of total starch and amylopectin in ‘312-1' were similar.They were all accumulated gradually with fruit development,and the peak value appeared in 40 days after pollination,then decreased slightly.While,the contents of total starch and amylopectin in ‘98-2' maintained the same level at the early stage of fruit development(5 days after pollination)without significant accumulation process.The contents of total starch and amylopectin in ‘312-1' were significantly higher than that in ‘98-2' during each fruit development stage.The amylose accumulation pattern in the 2 materials was similar,all maintaining at accumulated level at the early fruit development stage.However,the amylose contents in ‘312-1' at different fruit development stages were significantly or extremely significantly higher than that of ‘98-2'.In starch composition,amylopectin was dominant in the 2 materials.In addition to ISA Ⅲ,SSSⅡ and SBEⅡ genes,the GPT,AATP,PGM,AGPaseL,GBSSⅠ,SSS Ⅲ and SBEⅠ genes had similar expression trends during fruit development process of these 2 materials,and their expression quantities had large differences.Combining the amylopectin and amylopectin contents in fruits of these 2 materials,the paper speculated that GPT,AATP,AGPaseL,GBSSⅠ,SSSⅡ,SSS Ⅲ,SBEⅠ and SBEⅡ genes were key genes regulating starch accumulation in pumpkin fruit.
引文
高振宇,黄大年,钱前.2004.植物支链淀粉生物合成研究进展.植物生理与分子生物学学报,30(5):489-495.
孙守如,杨子琴,张菊平,翟庆慧,陈解放,巩振辉.2008.南瓜果实发育过程中糖代谢及相关酶活性的变化.西北农林科技大学学报,36(6):159-164.
王安君.2017.基于RNA-Seq的南瓜果实重要品质形成的代谢和分子调控研究[硕士论文].广州:暨南大学.
徐昌杰,陈文峻,陈昆松,张上隆.1998.淀粉含量测定的一种简便方法-碘显色法.生物技术,8(2):41-43.
徐丽珊,俞金龙,杨越安.2009.南瓜的含水量与淀粉含量的相关性研究.安徽农业科学,37(13):5928-5930.
杨金华,毕晨光,宋文昌.1992.半粒稻米直链淀粉测定方法的研究及应用.作物学报,18(5):366-372.
尹玲,王长林,王迎杰,向成钢,陈花.2012.南瓜品质的感官、质构及生化分析.食品科学,34(5):26-30.
郑义,陆维忠,马鸿翔.2009.植物淀粉生物合成的研究进展.江苏农业科学,(6):13-16.
Ball S G,Morell M K.2003.From bacterial glycogen to starch:understanding the biogenesis of the plant starch granule.Annual Review of Plant Biology,54(1):207-233.
Ferriol M,PicóB.2008.Pumpkin and Winter Squash//Prohens J,Nuez F.Handbook of plant breeding,vegetables I.New York:Springer:317-349.
Gentry M S,Brewer M K,Vander Kooi C W.2016.Structural biology of glucan phosphatases from humans to plants.Current Opinion in Structural Biology,40:62-69.
Hurst P,Corrigan V,Hannan P,Lill R.1995.Storage rots,compositional analysis,and sensory quality of three cultivars of buttercup squash.New Zealand Journal of Experimental Agriculture,23(1):89-95.
Kozlov S S,Blennow A,Krivandin A V,Yuryev V P.2007.Structural and thermodynamic properties of starches extracted from GBSS and GWD suppressed potato lines.International Journal of Biological Macromolecules,40(5):449-460.
Nakkanong K,Yang J H,Zhang M F.2012.Starch accumulation and starch related genes expression in novel inter-specific inbred squash line and their parents during fruit development.Scientia Horticulturae,136(2):1-8.
Otani M,Hamada T,Katayama K,Kitahara K,Kim S H,Takahata Y.2007.Inhibition of the gene expression for granule-bound starch synthase I by RNA interference in sweet potato plants.Plant Cell Reports,26(10):1801-1807.
Raemakers K,Schreuder M,Suurs L,Verhorst H F,Vincken J P,Vetten N D,Jacobsen E,Visser R G F.2005.Improved cassava starch by antisense inhibition of granule-bound starch synthase I.Molecular Breeding,16(2):163-172.
Stevenson D G,Yoo S H,Hurst P L,Jane J L.2005.Structural and physicochemical characteristics of winter squash(Cucurbita maxima D.)fruit starches at harvest.Carbohydrate Polymers,59(2):153-163.
Stitt M,Lunn J,Usadel B.2010.Arabidopsis and primary photosynthetic metabolism-more than the icing on the cake.Plant Journal,61(6):1067-1091.
Tetlow I,Liu F,Emes M.2008.Functional interactions between starch synthases and branching enzymes of cereal endosperms.Comparative Biochemistry and Physiology Part A:Molecular&Integrative Physiology,150(3):S194-S195.
Toyosawa Y,Kawagoe Y,Matsushima R,Crofts N,Ogawa M,Fukuda M,Kumamaru T,Okazaki Y,Kusano M,Saito K,Toyooka K,Sato M,Ai Y,Jane J L,Nakamura Y,Fujita N.2016.Deficiency of starch synthaseⅢa andⅣb alters starch granule morphology from polyhedral to apherical in rice endosperm.Plant Physiology,170(3):1255.
Vu V V,Beeson W T,Span E A,Farquhar E R,Marletta M A.2014.A family of starch-active polysaccharide monooxygenases.Proceedings of the National Academy of Sciences of the United States of America,111(38):13822-13827.
Wyatt L E,Strickler S R,Mueller L A,Michael M.2016.Comparative analysis of Cucurbita pepo metabolism throughout fruit development in acorn squash and oilseed pumpkin.Horticulture Research,3:16045.
Yun M S,Umemoto T,Kawagoe Y.2011.Rice debranching enzyme isoamylase3 facilitates starch metabolism and affects plastid morphogenesis.Plant and Cell Physiology,52(6):1068-1082.
Zeeman S C,Kossmann J,Smith A M.2010.Starch:its metabolism,evolution,and biotechnological modification in plants.Annual Review of Plant Biology,61(1):209-234.
孙守如,杨子琴,张菊平,翟庆慧,陈解放,巩振辉.2008.南瓜果实发育过程中糖代谢及相关酶活性的变化.西北农林科技大学学报,36(6):159-164.
王安君.2017.基于RNA-Seq的南瓜果实重要品质形成的代谢和分子调控研究[硕士论文].广州:暨南大学.
徐昌杰,陈文峻,陈昆松,张上隆.1998.淀粉含量测定的一种简便方法-碘显色法.生物技术,8(2):41-43.
徐丽珊,俞金龙,杨越安.2009.南瓜的含水量与淀粉含量的相关性研究.安徽农业科学,37(13):5928-5930.
杨金华,毕晨光,宋文昌.1992.半粒稻米直链淀粉测定方法的研究及应用.作物学报,18(5):366-372.
尹玲,王长林,王迎杰,向成钢,陈花.2012.南瓜品质的感官、质构及生化分析.食品科学,34(5):26-30.
郑义,陆维忠,马鸿翔.2009.植物淀粉生物合成的研究进展.江苏农业科学,(6):13-16.
Ball S G,Morell M K.2003.From bacterial glycogen to starch:understanding the biogenesis of the plant starch granule.Annual Review of Plant Biology,54(1):207-233.
Ferriol M,PicóB.2008.Pumpkin and Winter Squash//Prohens J,Nuez F.Handbook of plant breeding,vegetables I.New York:Springer:317-349.
Gentry M S,Brewer M K,Vander Kooi C W.2016.Structural biology of glucan phosphatases from humans to plants.Current Opinion in Structural Biology,40:62-69.
Hurst P,Corrigan V,Hannan P,Lill R.1995.Storage rots,compositional analysis,and sensory quality of three cultivars of buttercup squash.New Zealand Journal of Experimental Agriculture,23(1):89-95.
Kozlov S S,Blennow A,Krivandin A V,Yuryev V P.2007.Structural and thermodynamic properties of starches extracted from GBSS and GWD suppressed potato lines.International Journal of Biological Macromolecules,40(5):449-460.
Nakkanong K,Yang J H,Zhang M F.2012.Starch accumulation and starch related genes expression in novel inter-specific inbred squash line and their parents during fruit development.Scientia Horticulturae,136(2):1-8.
Otani M,Hamada T,Katayama K,Kitahara K,Kim S H,Takahata Y.2007.Inhibition of the gene expression for granule-bound starch synthase I by RNA interference in sweet potato plants.Plant Cell Reports,26(10):1801-1807.
Raemakers K,Schreuder M,Suurs L,Verhorst H F,Vincken J P,Vetten N D,Jacobsen E,Visser R G F.2005.Improved cassava starch by antisense inhibition of granule-bound starch synthase I.Molecular Breeding,16(2):163-172.
Stevenson D G,Yoo S H,Hurst P L,Jane J L.2005.Structural and physicochemical characteristics of winter squash(Cucurbita maxima D.)fruit starches at harvest.Carbohydrate Polymers,59(2):153-163.
Stitt M,Lunn J,Usadel B.2010.Arabidopsis and primary photosynthetic metabolism-more than the icing on the cake.Plant Journal,61(6):1067-1091.
Tetlow I,Liu F,Emes M.2008.Functional interactions between starch synthases and branching enzymes of cereal endosperms.Comparative Biochemistry and Physiology Part A:Molecular&Integrative Physiology,150(3):S194-S195.
Toyosawa Y,Kawagoe Y,Matsushima R,Crofts N,Ogawa M,Fukuda M,Kumamaru T,Okazaki Y,Kusano M,Saito K,Toyooka K,Sato M,Ai Y,Jane J L,Nakamura Y,Fujita N.2016.Deficiency of starch synthaseⅢa andⅣb alters starch granule morphology from polyhedral to apherical in rice endosperm.Plant Physiology,170(3):1255.
Vu V V,Beeson W T,Span E A,Farquhar E R,Marletta M A.2014.A family of starch-active polysaccharide monooxygenases.Proceedings of the National Academy of Sciences of the United States of America,111(38):13822-13827.
Wyatt L E,Strickler S R,Mueller L A,Michael M.2016.Comparative analysis of Cucurbita pepo metabolism throughout fruit development in acorn squash and oilseed pumpkin.Horticulture Research,3:16045.
Yun M S,Umemoto T,Kawagoe Y.2011.Rice debranching enzyme isoamylase3 facilitates starch metabolism and affects plastid morphogenesis.Plant and Cell Physiology,52(6):1068-1082.
Zeeman S C,Kossmann J,Smith A M.2010.Starch:its metabolism,evolution,and biotechnological modification in plants.Annual Review of Plant Biology,61(1):209-234.