[1]曹沛沛,毛雅超,刘涛,等.菊花Cm14-3-3v基因的克隆及表达分析[J].南京农业大学学报,2017,40(5):820-826.[doi:10.7685/jnau.201612048]
 CAO Peipei,MAO Yachao,LIU Tao,et al.Cloning and expression analysis of Cm14-3-3v gene in chrysanthemum[J].Journal of Nanjing Agricultural University,2017,40(5):820-826.[doi:10.7685/jnau.201612048]
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菊花Cm14-3-3v基因的克隆及表达分析()
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《南京农业大学学报》[ISSN:1000-2030/CN:32-1148/S]

卷:
40卷
期数:
2017年5期
页码:
820-826
栏目:
OA栏目
出版日期:
2017-09-20

文章信息/Info

Title:
Cloning and expression analysis of Cm14-3-3v gene in chrysanthemum
作者:
曹沛沛 毛雅超 刘涛 陈发棣 房伟民 陈素梅 蒋甲福
南京农业大学园艺学院, 江苏 南京 210095
Author(s):
CAO Peipei MAO Yachao LIU Tao CHEN Fadi FANG Weimin CHEN Sumei JIANG Jiafu
College of Horticulture, Nanjing Agricultural University, Nanjing 210095, China
关键词:
菊花Cm14-3-3v基因蛋白结构胁迫基因表达
Keywords:
chrysanthemumCm14-3-3v geneprotein structurestressgene expression
分类号:
S682.1+1
DOI:
10.7685/jnau.201612048
摘要:
[目的]本文旨在克隆菊花‘神马’Cm14-3-3v基因,并初步分析其表达特征,为其功能研究奠定基础。[方法]从菊花‘优香’转录组库中获得编码拟南芥同源基因14-3-3v序列信息,根据NCBI比对确定基因的cDNA全长并进行克隆,再利用生物信息学方法对该蛋白的功能和活性位点进行预测,同时利用实时定量PCR分析了在非生物胁迫条件下该基因的表达变化。[结果]Cm14-3-3v的ORF序列长774 bp,编码257个氨基酸;亚细胞定位表明Cm14-3-3v定位在细胞质和细胞核;生物信息学分析表明该蛋白有19个Ser、7个Thr和7个Tyr潜在的磷酸化位点,在N-端有5个蛋白结合区;表达模式分析显示,Cm14-3-3v基因在根、茎、叶、花中均有表达,且受脱落酸(ABA)、茉莉酸甲酯(MeJA)、盐和冷胁迫的诱导表达。[结论]本文克隆得到菊花Cm14-3-3v基因,表达模式分析表明其受ABA、MeJA、盐和冷胁迫诱导表达。
Abstract:
[Objectives]We cloned chrysanthemum ‘Jimba’ Cm14-3-3v gene and analyzed its expression characteristics,which provided the foundation for functional research.[Methods]The sequence encoding the 14-3-3v was obtained based on the annotation of genes from a RNA seq library of chrysanthemum,and the full length cDNA was cloned with RT-PCR. The bioinformatics method was used to predict the function and the characteristics of the protein,and the expression levels of the gene under different abiotic stresses were analyzed by qRT-PCR.[Results]The ORF of Cm14-3-3v was 774 bp,encoding 257 amino acids. Subcellular localization analysis showed that Cm14-3-3v was located in the cytoplasm and nucleus. The protein had 19 Ser,7 Thr and 7 Tyr potential phosphorylation sites,5 protein-binding regions at the N-terminal. The expression pattern analysis showed that the Cm14-3-3v gene was expressed in roots,stems,leaves,flowers,and was induced by abscissic acid(ABA),methyl jasmonate(MeJA),salt and cold stress.[Conclusions]The expression of Cm14-3-3v gene in chrysanthemum was cloned,its expression was induced by ABA,MeJA,salt,cold stress.

参考文献/References:

[1] 成丽娜,魏倩,Imtiaz M,等. 转基因育种技术在菊花性状改良中的应用进展[J]. 园艺学报,2013,40(9):1813-1825. Cheng L N,Wei Q,Imtiaz M,et al. Progress of application of gene transfer technology in chrysanthemum breeding traits improvement[J]. Journal of Horticulture,2013,40(9):1813-1825(in Chinese with English abstract).
[2] Sehnke P C,Delille J M,Ferl R J. Consummating signal transduction:the role of 14-3-3 proteins in the completion of signal-induced transitions in protein activity[J]. Plant Cell,2002,14(81):S339-S354.
[3] Chaudhri M,Scarabel M,Aitken A. Mammalian and yeast 14-3-3 isoforms form distinct patterns of dimers in vivo[J]. Biochemicaland Biophysical Research Communications,2003,300(3):679-685.
[4] Bunney T D,van Walraven H S,de Boer A H. 14-3-3 protein is a regulator of the mitochondrial and chloroplast ATP synthase[J]. Proc Natl Acad Sci USA,2001,98(7):42-49.
[5] Sehnke P C,Henry R,Cline K,et al. Interaction of a plant 14-3-3 protein with the signal peptide of a thylakoid-targeted chloroplast precursor protein and the presence of 14-3-3 isoforms in the chloroplast stroma[J]. Plant Physiology,2000,122(1):235-242.
[6] Chen F,Li Q,Sun L,et al. The rice14-3-3 gene family and its involvement in responses to biotic and abiotic stress[J]. DNA Res,2006,13(2):53-63.
[7] Song A,Li P,Jiang J,et al. Phylogenetic and transcription analysis of chrysanthemum WRKY transcription factors[J]. International Journal of Molecular Sciences,2014,15(8):14442-14455.
[8] Curtis M D,Grossniklaus U. A gateway cloning vector set for high-throughput functional analysis of genes in planta[J]. Plant Physiology,2003,133(2):462-469.
[9] Agrawal V D,Agrawal P. The 14-3-3 proteins of Arabidopsis regulate root growth and chloroplast development as components of the photosensory system[J]. Journal of Experimental Botany,2012,63(8):3061-3070.
[10] Fulgosi H,Soll J,Maraschin S D F,et al. 14-3-3 proteins and plant development[J]. Plant Molecular Biology,2002,50(6),1019-1029.
[11] Jarillo J A,Capel J,Leyva A,et al. Two related low-temperature-inducible genes of Arabidopsis encode proteins showing high homology to 14-3-3 proteins,a family of putative kinase regulators[J]. Plant Molecular Biology,1994,25(4):693-704.
[12] Merlot S,Leonhardt N,Fenzi F,et al. Constitutive activation of a plasma membrane H+-ATPase prevents abscisic acid-mediated stomatal closure[J]. Embo Journal,2007,26(13):3216-3226.
[13] Schoonheim P J,Sinnige M P,Casaretto J A,et al. 14-3-3 adaptor proteins are intermediates in ABA signal transduction during barley seed germination[J]. Plant Journal,2007,49(2):289-301.
[14] Gökirmak T,Paul A L,Ferl R J. Plant phosphopeptide-binding proteins as signaling mediators[J]. Current Opinion in Plant Biology,2007,13(5):527-532.
[15] Finni C,Andersen C H,Borch J,et al. Do 14-3-3 proteins and plasma membrane H+-ATPases interact in the barley epidermis in response to the barley powdery mildew fungus?[J]. Plant Molecular Biology,2002,49(2):137-147.
[16] Rosenquist M,Alsterfjord M,Larsson C,et al. Data mining the Arabidopsis genome reveals fifteen 14-3-3 genes. Expression is demonstrated for two out of five novel genes[J]. Plant Physiology,2001,127(1):142-149.
[17] Lapointe G,Luckevich M D,Séguin A. Investigation on the induction of 14-3-3 in white spruce[J]. Plant Cell Reports,2001,20(1):79-84.
[18] Lapointe G,Luckevich M D,Cloutier M,et al. 14-3-3 gene family in hybrid poplar and its involvement in tree defence against pathogens[J]. Journal of Experimental Botany,2001,52(359):1331-1338.
[19] Seehaus K,Tenhaken R. Cloning of genes by mRNA differential display induced during the hypersensitive reaction of soybean after inoculation with Pseudomonas syringae pv. glycinea[J]. Plant Molecular Biology,1998,38(6):1225-1234.
[20] Tsuji H,Nakamura H,Taoka K,et al. Functional diversification of FD transcription factors in rice,components of florigen activation complexes[J]. Plant and Cell Physiology,2013,54(3):385-397.
[21] Kim H S,Oh J M,Luan S,et al. Cold stress causes rapid but differential changes in properties of plasma membrane H(+)-ATPase of camelina and rapeseed[J]. Journal of Plant Physiology,2013,170(9):828-837.
[22] Mayfield J D,Folta K M,Paul A L,et al. The 14-3-3 proteins μ and υ influence transition to flowering and early phytochrome response[J]. Plant Physiology,2007,145(4):1692-1702.
[23] Adams E,Diaz C,Hong J P,et al. 14-3-3 proteins participate in light signaling through association with PHYTOCHROME INTERACTING FACTORs[J]. International Journal of Molecular Sciences,2014,15(12):22801-22814.

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备注/Memo

备注/Memo:
收稿日期:2016-12-30。
基金项目:国家自然科学基金项目(31572159);江苏省"双创计划"人才资助项目
作者简介:曹沛沛,硕士研究生。
通信作者:蒋甲福,教授,主要从事菊花遗传育种与分子生物学研究,E-mail:jiangjiafu@njau.edu.cn
更新日期/Last Update: 2017-09-23