MIAO Hong-xia,JIN Zhi-qiang,SUN Pei-guang,et al.Cloning and Expression Analysis of MaGBSSI Gene Family Members of Banana Fruit[J].Northern Horticulture,2014,38(20):115-120.
香蕉果实MaGBSSI基因家族的克隆及表达分析
- Title:
- Cloning and Expression Analysis of MaGBSSI Gene Family Members of Banana Fruit
- 文章编号:
- 1001-0009(2014)20-0115-06
- Keywords:
- banana; fruit; MaGBSSI gene; clone; expression analysis
- 分类号:
- S 668.103.6
- 文献标志码:
- A
- 摘要:
- 以“巴西”蕉(Musa acuminata L.AAA group cv.Brazilian)为试材,利用同源克隆法获得4个颗粒结合淀粉合成酶I(Granule-Bound Starch Synthase I, GBSSI)基因家族成员的cDNA全长,运用MEGA 5.05软件进行聚类分析,并通过quantitative real-time PCR(qPCR)技术检测MaGBSSI基因家族成员在香蕉果实不同发育时期及成熟阶段和不同组织中的表达情况。结果表明:香蕉4个MaGBSSI基因家族成员MaGBSSI-1、MaGBSSI-2、MaGBSSI-3、MaGBSSI-4的cDNA全长分别为1 851、1 851、675、1 845 bp,登录号分别为KF512020、KF512021、KF512022、KF512023。聚类分析发现,香蕉MaGBSSI基因家族与其它植物GBSSI基因同源性达65%。qPCR分析发现,MaGBSSI-1、MaGBSSI-2、MaGBSSI-4在香蕉根、球茎、叶片和苞片等营养器官中明显上调表达,而MaGBSSI-3在花、果皮、果肉等生殖器官中表达量比较高,在根、球茎及叶片中几乎不表达。香蕉果实发育过程中,MaGBSSI-1、MaGBSSI-2、MaGBSSI-4在0~30 d明显上调表达,而MaGBSSI-3在香蕉果实发育30~60 d表达量比较高,在0~30 d几乎不表达,且随着果实的成熟,MaGBSSI-3表达量逐渐下降。研究结果为阐明香蕉果实的直链淀粉生物合成及降解机制并对其进行表达调控奠定了基础。
- Abstract:
- Taking Musa acuminata L.AAA group cv.Brazilian as material,PCR amplification method of homologous gene was used to clone the cDNA full length of Granule-Bound Starch Synthase I (GBSSI) gene family in banana.Then,the cluster analysis of the gene family was performed by MEGA 5.05 software and its expression patterns in different tissues and different developmentals and harvest periods were tested by quantitative real-time PCR (qPCR).The results showed that the cDNA full length of four MaGBSSI gene family members,i.e.MaGBSSI-1,MaGBSSI-2,MaGBSSI-3 and MaGBSSI-4 were 1 851 bp,1 851 bp,675 bp and 1 845 bp,respectively.The accession number of MaGBSSI-1,MaGBSSI-2,MaGBSSI-3 and MaGBSSI-4 were KF512020,KF512021,KF512022 and KF512023,respectively.Cluster analysis showed that the MaGBSSI gene family from banana shared 65% amino acid sequence identifies with GBSSI gene from other plant.Tissue-specific expression of the MaGBSSI gene family members were detected using qPCR.MaGBSSI-1,MaGBSSI-2,and MaGBSSI-4 were up-regulated in vegetative tissue such as root,stem,leaf,and bract.In contrast,MaGBSSI-3 was highly expressed in reproductive tissues such as flower,peel,and pulp,but was weakly expressed in root,stem,and leaf.Expression levels of MaGBSSI-1,MaGBSSI-2,MaGBSSI-4,MaGBSSII-1,and MaGBSSII-2 at earlier stages of banana development (from 0 d to 30 d) were higher than the later stages (from 30 d to 60 d). In contrast,MaGBSSI-3 was weakly expressed at the early stages but was highly up-regulated at 50 d of development,while the expression level of MaGBSSI-3 was gradually decreased with the ripening of banana fruit.The results had laid the foundation to illustrate the amylose biosynthesis and degradation mechanism and regulate the expression of MaGBSSI gene family members of banana fruit.
参考文献/References:
[1]D’Hont A,Denoeud F,Aury J,et al.The banana (Musa acuminata) genome and the evolution of monocotyledonous plants [J].Nature,2012,488:213-217.
[2]Aparicio-Saguilan A,Osorio-Díaz P,Agama-Acevedo E,et al.Tortilla added with unripe banana and cassava flours:chemical composition and starch digestibility [J].CyTA-J Food,2013,11(1):90-95.
[3]Utrilla-Coello R G,Rodríguez-Huezo M E,Carrillo-Navas H,et al.In vitro digestibility,physicochemical,thermal and rheological properties of banana starches [J].Carbohydr Polym,2014,101:154-162.
[4]Krishnan H B,Chen M.Identification of an abundant 56 kDa protein implicated in food allergy as granule-bound starch synthase [J].J Agric Food Chem,2013,61(22):5404-5409.
[5]Vrinten P L,Nakamura T.Wheat granule-bound starch synthase I and II are encoded by separate genes that are expressed in different tissues [J].Plant Physiol,2000,122(1):255-264.
[6]Dian W,Jiang H,Chen Q,et al.Cloning and characterization of the granule-bound starch synthase II gene in rice:gene expression is regulated by the nitrogen level,sugar and circadian rhythm [J].Planta,2003,218:261-268.
[7]Hylton C M,Denyer K,Keeling P L,et al.The effect of waxy mutations on the granule-bound starch synthases of barley and maize endosperms [J].Planta,1996,198:230-237.
[8]Dry I,Smith A,Edwards A,et al.Characterization of cDNAs encoding two isoforms of granule-bound starch synthase which show differential expression in developing storage organs of pea and potato [J].Plant J,1992,2(2):193-202.
[9]Li Z,Li D,Du X,et al.The barley amo1 locus is tightly linked to the starch synthase IIIa gene and gegatively regulates expression of granule-bound starch synthetic genes [J].J Exp Bot,2011,62(14):5217-5231.
[10]Park Y J,Nemoto K,Nishikawa T,et al.Genetic diversity and expression analysis of granule bound starch synthase I gene in the new world grain amaranth (Amaranthus cruentus L.)[J].J Cereal Sci,2011,53(3):298-305.
[11]Cheng J,Khan M A,Qiu W M,et al.Diversification of genes encoding granule-bound starch synthase in monocots and dicots is marked by multiple genome-wide duplication events [J].PloS One,2012,7(1):e30088.
[12]罗明,肖月华,侯磊,等.棉花LM结构域基因(GhLM1)的克隆与表达分析[J].遗传学报,2003,30(2):175-182.
[13]Xu B Y,Su W,Liu J H,et al.Differentially expressed cDNAs at the early stage of banana ripening identified by suppression subtractive hybridization and cDNA microarray [J].Planta,2007,226:529-539. [14]Livak K J,Schimittgen T D.Analysis of relative gene expression data using real-time quantitative PCR and the 2-△△CT Method [J].Methods,2001,25(4):402-408.
[15]Biselli C,Cavalluzzo D,Perrini R,et al.Improvement of marker-based predictability of apparent amylose content in japonica rice through GBSSI allele mining [J].Rice,2014,7:1-18.
[16]Juárez-García E,Agama-Acevedo E,Gómez-Montiel N O,et al.Proteomic analysis of the enzymes involved in the starch biosynthesis of maize with different endosperm type and characterization of the starch [J].J Sci Food Agr,2013,93(11):2660-2668.
[17]Kimura T,Saito A.Heterogeneity of poly (A) sites in the granule-bound starch synthase I gene in sweet potato (Ipomoea batatas (L.) Lam.)[J].Biosci Biotech Bioch,2010,74(3):667-669.
[18]Park Y J,Nemoto K,Nishikawa T,et al.Molecular cloning and characterization of granule bound starch synthase I cDNA from a grain amaranth (Amaranthus cruentus L.)[J].Breeding Sci,2009,59(4):351-360. [19]Soares C A,Peroni-Okita F H G,Cardoso M B,et al.Plantain and banana starches:granule structural characteristics explain the differences in their starch degradation patterns [J].J Agric Food Chem,2011,59(12):6672-6681.
[20]Sarawong C,Schoenlechner R,Sekiguchi K,et al.Effect of extrusion cooking on the physicochemical properties,resistant starch,phenolic content and antioxidant capacities of green banana flour [J].Food Chem,2014,143:33-39.
[21]苗红霞,金志强,刘伟鑫,等.香蕉果实抗性淀粉变化及其与其他类型淀粉相关性分析[J].中国农业科学,2013,46(24):180-5187.
[22]Miao H X,Sun P G,Liu W X,et al.Identification of genes encoding granule-bound starch synthase involved in amylose metabolism in banana fruit [J].PLoS One,2014,9(2):e88077.
相似文献/References:
[1]金方伦,敖学熙,张发维,等.中华猕猴桃结果蔓上果实节位与生长发育动态分析[J].北方园艺,2014,38(13):15.
JIN Fang-lun,AO Xue-xi,ZHANG Fa-wei,et al.Analysis on the Growth and Development Dynamic of the Kiwi Fruit[J].Northern Horticulture,2014,38(20):15.
[2]金方伦,张发维,周光萍,等.纽荷尔脐橙在黔北地区的果实发育动态分析[J].北方园艺,2014,38(11):20.
JIN Fang-lun,ZHANG Fa-wei,ZHOU Guang-ping,et al.Analysis on the Fruit Growth Dynamic of Newhall Navel Orange in North of Guizhou[J].Northern Horticulture,2014,38(20):20.
[3]金方伦,冯世华,张发维,等.不同留果量对中华猕猴桃后熟期果实品质的影响[J].北方园艺,2014,38(10):26.
JIN Fang-lun,FENG Shi-hua,ZHANG Fa-wei,et al.Effect of Different Fruit Retaining on Fruit Quality of Kiwi at Maturation Period[J].Northern Horticulture,2014,38(20):26.
[4]王 强,张茂君,闫兴凯,等.不同贮藏期“寒香”梨果实香气成分的GC-MS分析[J].北方园艺,2013,37(22):32.
WANG Qiang,ZHANG Mao-jun,YAN Xing-kai,et al.Analysis of GC-MS on Fruit Aroma Components of ‘Hanxiang’ Pear During Different Storage Periods[J].Northern Horticulture,2013,37(20):32.
[5]常立儒.‘山梨’果实和叶片发育期有机酸含量的变化及其相关性研究[J].北方园艺,2014,38(04):26.
CHANG Li-ru.Correlation Analysis of Changes in the Organic Acid Contents of Sorb During Fruit and Leaf Development[J].Northern Horticulture,2014,38(20):26.
[6]解贝贝,沙广利,戴洪义,等.六种无融合生殖砧木对‘嘎拉’苹果果实大小和品质的影响[J].北方园艺,2013,37(17):23.
XIE Bei-bei,SHA Guang-li,DAI Hong-yi,et al.Effect of Six No Fusion Reproductive Rootstocks on the Fruit Size and Quality of ‘Gala’ Apple[J].Northern Horticulture,2013,37(20):23.
[7]周淑香,符亚茹,王 超,等.西藏光核桃果实及种子表型性状变异研究[J].北方园艺,2013,37(23):38.
ZHOU Shu-xiang,FU Ya-ru,WANG Chao,et al.Study on Phenotypic Variation of Seed and Fruit on Prunus miakoehne in Tibet[J].Northern Horticulture,2013,37(20):38.
[8]郑丽屏,蔡 平,王利芬,等.苏州光福不同杨梅品种果实品质分析[J].北方园艺,2014,38(09):38.
ZHENG Li-ping,CAI Ping,WANG Li-fen,et al.Analysis of Fruit Quality of the Different Myrica rubra Varieties in Suzhou Guangfu[J].Northern Horticulture,2014,38(20):38.
[9]姜喜,马宝蕴,陈加利,等.香梨生长季果实和叶片矿质元素含量的变化[J].北方园艺,2013,37(12):15.
JIANG Xi,MA Bao-yun,CHEN Jia-li,et al.Variation of Mineral Element Content in Fragrant Pear Fruits and Leaves During Its Development[J].Northern Horticulture,2013,37(20):15.
[10]张欣欣,郭军战,郭冬梅.果桑不同品种生长结实特性比较研究[J].北方园艺,2013,37(01):21.
ZHANG Xin-xin,GUO Jun-zhan,GUO Dong-mei.Study on the Characteristics of Growth and Fruiting of Different Fruit Mulberry Varieties[J].Northern Horticulture,2013,37(20):21.
备注/Memo
第一作者简介:苗红霞(1982-),女,博士,助理研究员,研究方向为香蕉分子生物学。E-mail:hxmrain@163.com.
责任作者:徐碧玉(1962-),女,博士,研究员,博士生导师,研究方向为香蕉遗传育种。E-mail:biyuxu@126.com.
基金项目:“十二五”农村领域国家科技计划资助项目(2011AA10020605);现代农业产业技术体系建设专项资金资助项目(CARS-32);海南省自然科学基金资助项目(314116);中央级公益性科研院所基本科研业务费专项资金资助项目(ITBB140205)。
收稿日期:2014-06-24