miR171家族保守性分析及其在香蕉叶片不同温度处理下的表达分析
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摘要
[目的]研究三明野生蕉和天宝栽培蕉在不同温度处理后mi R171家族成员在叶片中的表达情况,初步探索香蕉mi R171家族成员的抗寒机制。[方法]采用生物信息学分析和实时荧光定量PCR(q PCR)的方法,对香蕉中mi R171家族成员进行研究。[结果]序列比对结果表明,mi R171前体与成熟体序列间有2个保守性较高的重叠区,推测mi R171成熟体可能来自这2个区域。实时荧光定量PCR结果表明,三明野生蕉的mi R171b的相对表达量在0℃时达到最高,而天宝栽培蕉mi R171b则在4℃处理时表达量达到最高;mi R171c和mi R171c*在野生蕉和天宝蕉中具有相似的表达模式,其表达量都在4℃达到最高。[结论]mi R171家族成员可能具有多重调节功能,其家族成员mi R171b,mi R172c和mi R171c*可能在香蕉冷胁迫中发挥重要作用。
[Objective] To explore the function of mi R171 family members in cold resistance mechanism of Sanming'banana and Tianbao'banana,the expression pattern of the mi R171 family of banana was investigated in leaves after different temperature stress treatment. [Method]Bioinformatics analysis and quantitative real-time PCR methods were used to study mi R171 family members in bananas. [Result]Sequence alignment showed that there were two highly conserved regions between the mi R171 precursor sequences and the mature sequences,suggesting that the mi R171 mature sequences may mainly originated from these two regions. Quantitative real-time PCR analysis showed that mi R171 b in Sanming'banana had the highest relative expression at 0℃,while Tianbao'banana mi R171 b reached the highest expression at 4℃;mi R171 c and mi R171 c* had similar expression patterns in Sanming'banana and Tianbao'banana,and their expression level reached the peak at 4℃. [Conclusion] mi R171 family may have multiple regulatory functions,and mi R171 b,mi R171 c,and mi R171 c* may play important roles in the cold stress of bananas.
[Objective] To explore the function of mi R171 family members in cold resistance mechanism of Sanming'banana and Tianbao'banana,the expression pattern of the mi R171 family of banana was investigated in leaves after different temperature stress treatment. [Method]Bioinformatics analysis and quantitative real-time PCR methods were used to study mi R171 family members in bananas. [Result]Sequence alignment showed that there were two highly conserved regions between the mi R171 precursor sequences and the mature sequences,suggesting that the mi R171 mature sequences may mainly originated from these two regions. Quantitative real-time PCR analysis showed that mi R171 b in Sanming'banana had the highest relative expression at 0℃,while Tianbao'banana mi R171 b reached the highest expression at 4℃;mi R171 c and mi R171 c* had similar expression patterns in Sanming'banana and Tianbao'banana,and their expression level reached the peak at 4℃. [Conclusion] mi R171 family may have multiple regulatory functions,and mi R171 b,mi R171 c,and mi R171 c* may play important roles in the cold stress of bananas.
引文
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[2]CAR R IN GTO N J C,AMBR O S V.R ole of micro R N As in plant and animal development[J].Science,2003,301(5631):336-338.
[3]BAR TEL D P.Micro R N As:genomics,biogenesis,mechanism,and function[J].Cell,2004,116(2):281-297.
[4]LEE R C,FEIN BAUM R L,AMBR O S V.The C.elegans heterochronic gene lin-4 encodes small R N As with antisense complementarity to lin-14.[J].Cell,1993,75(5):843.
[5]KO ZO MAR A A,GR IFFITHSJO N ES S.mi R Base:integrating micro R N A annotation and deep-sequencing data[J].N ucleic Acids R esearch,2011,39(Database issue):D152.
[6]R EIN HAR T B J,WEIN STEIN E G,R HO ADES M W,et al.Micro R N As in plants[J].Genes&Development,2002,16(13):1616.
[7]YAN GLI,WEILI,YO U-XIN JIN.Computational identification of novel family members of micro R N A genes in Arabidopsis thaliana and Oryza sativa[J].生物化学与生物物理学报:英文版,2005,37(2):75-87.
[8]ABO O BAKER A A,TO MAN CAK P,PATEL N,et al.Drosophila micro R N As exhibit diverse spatial expression patterns during embryonic development[J].Proceedings of the N ational Academy of Sciences of the United States of America,2005,102(50):18017-18022.
[9]张曼曼.高山松.micro R N A171a的功能研究[D].新乡:河南师范大学,2014.
[10]LO N G W,MAI Y X,ZHAN G Y C,et al.Micro R N A171c-targeted SCL6-II,SCL6-III,and SCL6-IV genes regulate shoot branching in Arabidopsis.[J].Molecular Plant,2010,3(5):794-806.
[11]WU X M,LIU M Y,GE X X,et al.Stage and tissue-specific modulation of ten conserved mi R N As and their targets during somatic embryogenesis of Valencia sweet orange[J].Planta,2011,233(3):495-505.
[12]CAR MAN J G.EMBR YO GEN IC cells in plant tissue cultures:O ccurrence and behavior[J].Vitro Cellular&Developmental Biology,1990,26(8):746-753.
[13]ZHAN G J,ZHAN G S,HAN S,et al.Genome-wide identification of micro R N As in larch and stage-specific modulation of 11 conserved micro R N As and their targets during somatic embryogenesis.[J].Planta,2012,236(2):647-657.
[14]MAHALE B M,FAKR UDIN B,GHO SH S,et al.LN A mediated in situ hybridization of mi R 171 and mi R 397a in leaf and ambient root tissues revealed expressional homogeneity in response to shoot heat shock in Arabidopsis thaliana[J].Journal of Plant Biochemistry&Biotechnology,2014,23(1):93-103.
[15]曾友竞,林玉玲,崔彤彤,等.龙眼mi R 171家族进化特性及其表达分析[J].西北植物学报,2017,37(2):258-265.
[16]刘玉凤,王孝,陈海银,等.植物中mi R N A响应非生物胁迫研究进展[J].中国蔬菜,2017(2):23-29.
[17]张金梅.柑橘中冷胁迫相关mi R N As的克隆与鉴定[D].长沙:湖南农业大学,2011.
[18]周红玲,陈石,郑云云,等.香蕉寒害冻害及抗寒研究进展[J].现代农业科技,2017(4):57-58.
[19]李卫亮,李茂富,贺军虎,等.香蕉抗寒相关功能基因研究进展[J].分子植物育种,2015,13(5):1185-1192.
[20]祁君凤,冯仁军,程萍,等.香蕉叶片低温胁迫下差异表达c DN A消减文库的构建[J].热带生物学报,2010,1(2):110-113.
[21]SUN KAR R,ZHU J K.N ovel and stress-regulated micro R N As and other small R N As from Arabidopsis[J].Plant Cell,2004,16(8):2001-2019.
[22]MALLO R Y A C,VAUCHER ET H.Functions of micro R N As and related small R N As in plants[J].N ature Genetics,2006,38 Suppl(Suppl):S31.
[23]LLAVE C,CAR R IN GTO N J C.Cleavage of scarecrow-like m R N A targets directed by a class of arabidopsis mi R N A[J].Science,2002,297(5589):2053-2056.
[24]EMER Y J F,FLO YD S K,ALVAR EZ J,et al.R adial patterning of Arabidopsis shootsby class III HD-ZIP and KAN ADI genes.[J].Current Biology,2003,13(20):1768-1774.
[25]ZHAO S Z,SI Q H,ZHI M Y.Bioinformatic identification and expression analysis of new micro R N As from Medicago truncatula[J].Biochemical&Biophysical R esearch Communications,2008,374(3):538-542.
[26]薄维平,曾长英,宋顺,等.木薯耐寒相关micro R N A的差异表达分析[J].热带作物学报,2010,31(8):1260-1265.
[27]FAN T,LI X,YAN G W,et al.R ice osa-mi R 171c mediates phase change from vegetative to reproductive development and shoot apical meristem maintenance by repressing four O s HAM transcription factors[J].Plos O ne,2015,10(5):e0125833.
[28]王维,张玉娟,陈洁,等.植物逆境胁迫相关mi R N A研究进展[J].生物技术通报,2015,31(1):1-10.
[29]朱全武,范凯,谢艳兰,等.植物低温胁迫响应mi R N As及其在茶树抗寒研究中的应用[J].茶叶科学,2013(3):212-220.
[30]李世鹏.玉米自交系苗期冷胁迫mi R N A表达谱比较研究[D].长春:吉林大学,2016.
[31]熊雪梅.盐芥冷响应mi R N A表达模式分析及tsa-mi R 396功能初探[D].哈尔滨:东北林业大学,2014.