小麦条锈菌III型磷脂酰肌醇4-羟基激酶基因（PsPik1）
的功能分析
何付新;张阳;秦娟;马微;康振生;郭军
【期刊名称】《中国农业科学》 【年(卷),期】2015(000)016
【摘要】Objective]The objective of this study is to clone a gene PsPik1 encoding type III PtdIns 4-kinase from Puccinia striiformis f. sp. tritici (Pst) and characterize its function during Pst growth and development. [Method]The full-length cDNA of PsPik1 was obtained using RT-PCR. Bioinformatic methods were used to analyze molecular properties of PsPik1. Quantitative real-time PCR (qRT-PCR) was applied to examine the expression profiles of PsPik1 at twelve different development stages. The transcript level of PsPik1 was calculated by the 2-△△CT method with the EF1 gene of Pst as endogenous reference for normalization. BSMV-HIGS was employed to analyze the function of PsPik1 by knocking down its expression. Selected PsPik1 gene fragment was amplified by PCR from Pst cDNA using primers with restriction enzymes Not I and Pac I sites. Amplicon was ligated into the BSMVγvector generating BSMV:γ:PsPik1-as. The native BSMV:γ:0-as and BSMV:γ:TaPDS-as were used as negative and positive control, respectively. After inoculation with BSMV at the two-leaf stage, wheat seedlings were maintained in a growth chamber and examined for symptoms at regular intervals. After
小麦条锈菌III型磷脂酰肌醇4-羟基激酶基因(PsPik1)的功能分析
