玉米RCA基因克隆及表达分析.pdfVIP

玉米RCA基因克隆及表达分析 玉米RCA基因克隆及表达分析 印志同 印志同 ztyin@yzu.edu.cn 扬州大学农学院玉米遗传育种课题组 扬州大学农学院玉米遗传育种课题组 • 改善作物光合性能是提高产量的重要途径之一 • Rubisco activase (RCA) catalyzes the activation of Rubisco in vivo and plays a crucial role in regulating plant growth. RCA RCA是一种核编码的叶绿体蛋白，具有分子伴 侣的功能，在多数物种中含有α 和β 两种亚基。 • In maize, two β-isoform RCA genes (Zmrca1 and Zmrca2) with an identical open reading frame (ORF) were reported. 翻 译 43kD （β亚基） 水 解 41kD Vargas-Suarez et al., J Exp Bot,2004 Ayala-Ochoa et al., Biochimie 2004 玉米在热胁迫条件下产生45-46kD RCA多肽 Zoran Ristic et al.J Exp Bot,2009 Questions 1 玉米中是否存在RCA α亚基和基因？ 2 前人报道的45-46kD RCA多肽来源？ 3玉米RCA基因与产量的关系如何? RESULTS 1. Genomic Analysis and cDNA Cloning Revealed a New α- form RCA Gene and a Previously Reported β-form Gene 2.Zmrca1 and Zmrca3 Encoded two Different Maize Polypeptide Panel A: anti-Zmrca3 antibody. Panel B: anti-Zmrca1 antibody. Lane1:pET-30a-Zmrca1 Panel C: native Zmrca1 (lane1), lane 2: pET-30a-Zmrca3 native Zmrca3 (lane2), lane 3: empty E.coli leaf protein (lanes 3,4) lane 4: E. coli transformed with empty pET-30a lane 5: Arabidopsis lane 6: maize. Recombinant Native Zmrca3 Zmrca3 Recombinant Native Zmrca1 Zmrca1 MALDI-TOF-MS analysis 3.Zmrca1 and Zmrca3 Were Targeted to Chloroplast 4.Zmrca1 and Zmrca3 Showed Leaf-predominant and Cyclic Day/Night Expression Patterns 5. The Expression Levels of the Zmrca1 and Zmrca3 Were Correlated Wit