小麦三雌蕊突变体<em>TaAP1-3</em>基因的克隆及表达分析

doi:10.7668/hbnxb.2014.06.006

摘要/Abstract

摘要： 为探讨 TaAP1-3 基因在小麦三雌蕊性状形成中的作用,从小麦三雌蕊近等基因系CM28和CM28TP中克隆得到3个同源基因 TaAP1-3a、 TaAP1-3b 和 TaAP1-3c。通过碱基序列、氨基酸序列、聚类及实时荧光定量分析表明, TaAP1-3a、 TaAP1-3b 和 TaAP1-3c 基因cDNA 序列分别为1 210,1 208,1 199 bp,ORF分别为825,816,855 bp,分别编码274,271,284个氨基酸残基。 TaAP1-3a、 TaAP1-3b 和 TaAP1-3c 与中国春 TaAP1-3 的核苷酸序列相似度分别为96.02%,93.1%,93.56%,氨基酸序列相似性高达99%,95%,97%,并且与A类基因 TaAGL29、 OsMADS15、 ZAP1、 BM8、 EnWM8、 TaAP1-3 聚集到AP1/SUQA亚家族FUL2类群中。实时荧光定量分析表明, TaAP1-3a、 TaAP1-3b 和 TaAP1-3c 在CM28和CM28TP的3个发育时期的小穗中均有表达,但各个时期的表达量有明显差异。CM28中主要在二棱期-小花分化期表达,CM28TP中主要在雌雄蕊原基分化期表达。试验分析显示, TaAP1-3a、 TaAP1-3b 和 TaAP1-3c 可能具有上述A类基因相似的功能,其表达模式可能与小麦三粒/一粒性状形成相关。试验结果为进一步探讨该基因在小麦三雌蕊性状形成过程中的作用奠定了基础。

关键词: 小麦, 花器官发育, TaAP1-3, MADS-box基因, 基因表达

Abstract: To investigate the role of TaAP1-3 in the formation of wheat three pistils trait.Three homologous genes TaAP1-3a, TaAP1-3b and TaAP1-3c were cloned from wheat three pistils near-isogenic lines CM28 and CM28TP in this study.By nucleotide sequence,amino acid sequence,quantification and clustering analysis showed that the cloned cDNA of TaAP1-3a, TaAP1-3b and TaAP1-3c were 1 210,1 208,1 199 bp in length,respectively. TaAP1-3a, TaAP1-3b and TaAP1-3c genes respectively contained an 825,816,855 bp ORF which encode 274,271,284 amino acids.The cDNA sequence of TaAP1-3a, TaAP1-3b and TaAP1-3c with TaAP1-3 were respectively 96.02%,93.1% and 93.56% similarity.And the identities to TaAP1-3 in amino acids sequences were 99%,95%,97% respectively.Cluster analysis showed that TaAP1-3a, TaAP1-3b and TaAP1-3c were clustered into FUL2 clade,and showed greater similarity to A class function genes,such as, TaAGL29, OsMADS15, ZAP1, BM8, EnWM8E and TaAP1-3 genes.Real-time analysis showed the expression level of TaAP1-3a, TaAP1-3b and TaAP1-3c in CM28 and CM28TP were significantly different.For CM28,high expression level was shared in two ridge until floret differentiation (spikelet length 2-5 mm).However,the expression level was higher in the pistil and stamen primordium (spikelets length 5-7mm) of CM28TP.Experimental analysis indicated that the genes of TaAP1-3a, TaAP1-3b, TaAP1-3c may have similar function with A class function genes of above-mentioned,and the difference of expression pattern in CM28 and CM28TP may be bound up with three-pistil traits.The results make the foundation to further explore their roles in forming three-pistil traits in wheat.

Key words: Wheat, Floral organ development, TaAP1-3, MADS-box gene, Gene expression

中图分类号:

S512.1.1
Q78

王育伟, 彭正松, 杨在君, 魏淑红, 廖明莉, 赵欢, 杨会, 杨宇凤, 王清海. 小麦三雌蕊突变体TaAP1-3基因的克隆及表达分析[J]. 华北农学报, 2014, 29(6): 28-35. doi: 10.7668/hbnxb.2014.06.006.

WANG Yu-wei, PENG Zheng-song, YANG Zai-jun, WEI Shu-hong, LIAO Ming-li, ZHAO Huan, YANG Hui, YANG Yu-feng, WANG Qing-hai. Cloning and Expression Analysis of TaAP1-3 Genes of Three Pistils Mutation in Wheat[J]. ACTA AGRICULTURAE BOREALI-SINICA, 2014, 29(6): 28-35. doi: 10.7668/hbnxb.2014.06.006.

http://www.hbnxb.net/CN/Y2014/V29/I6/28

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小麦三雌蕊突变体TaAP1-3基因的克隆及表达分析

Cloning and Expression Analysis of TaAP1-3 Genes of Three Pistils Mutation in Wheat

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小麦三雌蕊突变体TaAP1-3基因的克隆及表达分析

Cloning and Expression Analysis of TaAP1-3 Genes of Three Pistils Mutation in Wheat

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