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 Ta AP1-3 Genes of Three Pistils Mutation in Wheat

作　　者：王育伟[1] 彭正松[1] 杨在君[1] 魏淑红[1] 廖明莉[1] 赵欢[1] 杨会[1] 杨宇凤 王清海[1] 

机构地区：[1]西华师范大学,西南野生动植物资源保护省部共建教育部重点实验室,四川南充637009

出　　处：《华北农学报》2014年第6期28-35,共8页Acta Agriculturae Boreali-Sinica

基　　金：国家自然科学基金项目(31301319);教育部重点项目(211164)

摘　　要：为探讨Ta AP1-3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Ta AP1-3a、Ta AP1-3b和Ta AP1-3c。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、Ta AP1-3b和Ta AP1-3c基因c DNA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。Ta AP1-3a、Ta AP1-3b和Ta AP1-3c与中国春Ta AP1-3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Ta AGL29、Os MADS15、ZAP1、BM8、En WM8、Ta AP1-3聚集到AP1/SUQA亚家族FUL2类群中。实时荧光定量分析表明,Ta AP1-3a、Ta AP1-3b和Ta AP1-3c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,Ta AP1-3a、Ta AP1-3b和Ta AP1-3c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。To investigate the role of Ta AP1-3 in the formation of wheat three pistils trait. Three homologous genes Ta AP1-3a,Ta AP1-3b and Ta AP1-3c were cloned from wheat three pistils near-isogenic lines CM28 and CM28 TP in this study. By nucleotide sequence,amino acid sequence,quantification and clustering analysis showed that the cloned c DNA of Ta AP1-3a,Ta AP1-3b and Ta AP1-3c were 1 210,1 208,1 199 bp in length,respectively.Ta AP1-3a,Ta AP1-3b and Ta AP1-3c genes respectively contained an 825,816,855 bp ORF which encode 274,271,284 amino acids. The c DNA sequence of Ta AP1-3a,Ta AP1-3b and Ta AP1-3c with Ta AP1-3 were respectively96. 02%,93. 1% and 93. 56% similarity. And the identities to Ta AP1-3 in amino acids sequences were 99%,95%,97% respectively. Cluster analysis showed that Ta AP1-3a,Ta AP1-3b and Ta AP1-3c were clustered into FUL2 clade,and showed greater similarity to A class function genes,such as,Ta AGL29,Os MADS15,ZAP1,BM8,EnWM8 E and Ta AP1-3 genes. Real-time analysis showed the expression level of Ta AP1-3a,Ta AP1-3b and Ta AP1-3c in CM28 and CM28 TP 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 CM28 TP. Experimental analysis indicated that the genes of Ta AP1-3a,Ta AP1-3b,Ta AP1-3c may have similar function with A class function genes of above-mentioned,and the difference of expression pattern in CM28 and CM28 TP may be bound up with three-pistil traits. The results make the founda-tion to further explore their roles in forming three-pistil traits in wheat.

关 键 词：小麦 花器官发育 TaAP1-3 MADS-BOX基因 基因表达 

分 类 号：S512.1[农业科学—作物学] Q78[生物学—分子生物学]