Rezapoor Habib, Aghdasi Mahnaz, Sadeghipoor Hamid Reza
Department of Biology, Faculty of Science, Golestan University, Gorgan, P.O.Box: 159, Iran.
Iran J Biotechnol. 2017 Mar;15(1):33-41. doi: 10.15171/ijb.1342.
TRR14 protein is a small protein, a member of a multigene family in Arabidopsis which was found as the fi rst protein during screening seedlings for their resistant to the trehalose sugar.
A number of TRR14-overexpressing plants were subjected to the characterization in the present research, among which, the associated morphological features and changes accompany growth pattern and photosynthesis related parameters.
gene was isolated from and cloned into the pBin-35S vector. Recombinant vector was transferred to the (Col-0) via using the Floral Dipping method. Seeds from the overexpressed (TRR14) and the Col-0 wild-type (WT) plants were shown on soil under long day conditions. Several measurements were then performed including determination of the fresh and dry weights, leaf area, chlorophyll a and b (Chl a and Chl b) content, Chl a/b ratio, total chlorophyll and carotenoids content, soluble and insoluble sugars content, total and soluble protein content, the Hill reaction rate, chlorophyll fl uorescence, as well as photorespiration rate. Meanwhile, the chloroplastic proteins were investigated by SDS-PAGE analysis.
TRR14 plants showed a signifi cant increase in fresh and dry weights, leaf area, and total and soluble protein content along with a signifi cant decrease in the insoluble sugar contents was observed in comparison to the WT plants. Chl a, Chl b, total chlorophyll content, Chl a/b ratio, carotenoids content, Hill reaction rate, and chlorophyll fl uorescence didn't show a signifi cant diff erence between TRR14 and WT plants. The SDS-PAGE gel electrophoresis of the chloroplastic proteins showed a thick band with a molecular mass of 25 kDa in -overexpressed plants, compared to the WT plants. Remarkably, photorespiration rate was decreased in TRR14 plants compared to WT plants.
The increased biomass of TRR14 transformed plants might be due to its ability in reducing photorespiration through concentrating CO in the leaf's intercellular spaces.
TRR14蛋白是一种小蛋白,是拟南芥中一个多基因家族的成员,它是在筛选对海藻糖具有抗性的幼苗过程中发现的首个蛋白。
在本研究中,对一些过表达TRR14的植株进行了特性分析,其中包括相关的形态特征以及伴随生长模式和光合作用相关参数的变化。
从[具体来源]中分离出基因并克隆到pBin - 35S载体中。通过花浸染法将重组载体利用农杆菌转入拟南芥(Col - 0)。将过表达TRR14的植株和Col - 0野生型(WT)植株的种子在长日照条件下播种于土壤中。然后进行了多项测定,包括鲜重和干重、叶面积、叶绿素a和b(Chl a和Chl b)含量、Chl a/b比值、总叶绿素和类胡萝卜素含量、可溶性和不溶性糖含量、总蛋白和可溶性蛋白含量、希尔反应速率、叶绿素荧光以及光呼吸速率的测定。同时,通过SDS - PAGE分析对叶绿体蛋白进行了研究。
与WT植株相比,TRR14植株的鲜重、干重、叶面积以及总蛋白和可溶性蛋白含量显著增加,同时不溶性糖含量显著降低。TRR14植株和WT植株之间的Chl a含量、Chl b含量、总叶绿素含量、Chl a/b比值、类胡萝卜素含量、希尔反应速率和叶绿素荧光没有显著差异。与WT植株相比,过表达TRR14的植株叶绿体蛋白的SDS - PAGE凝胶电泳显示出一条分子量为25 kDa的粗带。值得注意的是,与WT植株相比,TRR14植株的光呼吸速率降低。
TRR14转化植株生物量的增加可能是由于其通过在叶片细胞间隙浓缩CO₂来降低光呼吸的能力。