中国农业科学Issue(15)：3094-3102,9.DOI:10.3864/j.issn.0578-1752.2014.15.019
拟南芥PMRP的表达特性及功能分析
PMRP Expression Characteristics and Analysisof the Function inArabidopsis
摘要
Abstract
[Objective]The objective of this study is to analyze the expression characteristics ofPMRP (putative membrane related protein)gene (the biological funcion was unknown )inArabidopsis, and make clear of the regulation function of PMRP in Arabidopsis development process.[Method]Bioinformatics method was used to look for the genes which contain the same domain with PMRP in Arabidopsisand draw the phylogenetic tree. Real-time PCR technique was used to analyze the expression level of PMRP in root and stem tissues of 8 and 21 daysArabidopsis,compare the expression ofPMRP gene in the 1st, 2nd, 3rd and the 4th pairs of rosette leaves and cauline leaves in 21 daysArabidopsis, analyze the expression level ofPMRPin flower organs (such as sepals, stamens and petals) and seeds ofArabidopsis. The gain-of-function mutant of PMRP were obtained by constructing a vector ofPMRP gene driven by the 35S promoter of the cauliflower mosaic virus, and introduced it into wild typeArabidopsis Columbia, then tested the expression level of PMRP by RT-PCR technique. The regulating effect ofPMRP gene on the growth sites of leaves and stems was analyzed by observing the phenotype of the35S:PMRP transgenic plants. The function ofPMRP gene in differentiation of vascular xylem and phloem was observed using the paraffin sections of the stem transverse section in thePMRP over-expressing transgenicArabidopsis. The effect ofPMRP on the growth and developmentof Arabidopsis floral organ was studied through observation of the floral organ anatomy. The effect ofPMRP on fertility inArabidopsis thaliana was analyzed by observing the pod formation in thePMRP over-expressing transgenicArabidopsis.[Result]PMRP is a 411 amino acids protein which containing a START domain and have transmenbrane segments, there are 35 START proteins inArabidopsis. Real-time PCR analysis results showed that the expression level ofPMRP was higher in cauline leaves ( the relative expression ofPMRPwas about 2 935) and then the rosette leaves, the growth time of rosette leaves was longer, the more expression of PMRPwas tested (the relative expression ofPMRP in the 1st, 2nd, 3rd and 4th pairs of the rosette leaves were 1 650, 1 113, 734, and 507, respectively) , then the sepals floral tissues (PMRP relative expression quantity is about 937),PMRP had distribution in the stem, root and seed , but the relative expression amount was less than 270. The relative expression level ofPMRP in stamens was the lowest (about 64), far below the expression quantity ofPMRP in sepals (937). The expression level ofPMRP gene was increased with the increase of growth time, the relative expression ofPMRP were 154 and 222, respectively, in 8 and 21 d roots, and 200 and 264, respectively, in 8 and 21 d stems. The rosette leaves were emerged from the branches, stalk lodging susceptible, vascular had no obvious cambium and the xylem and phloem arranged in disorder, the filaments became shorter in floral organs, the number of pod decreased, and the fertility reduced.[Conclusion]The START domain is extremely conservative in function.PMRP was expressed in different organs ofA. thaliana, with the extension of time, the expression level ofPMRP was also increased. The level ofPMRP in the flower stamens was the lowest, oncePMRPwas over-expressed inArabidopsis floral organs, stamens dysplasia, resulting in reduced fertility.PMRP had important function in regulating the origination of leaves, the vascular differentiation and development of floral organs.关键词
拟南芥/PMRP/功能分析Key words
Arabidopsis/PMRP/function analysis引用本文复制引用
张昊,由诗东,高静,张海丽,李生辉,邢继红,王凤茹,董金皋..拟南芥PMRP的表达特性及功能分析[J].中国农业科学,2014,(15):3094-3102,9.基金项目
河北省自然科学基金 （）
