棉花八氢番茄红素脱氢酶GhPDS1基因的克隆与表达谱分析

摘要

Phytoene desaturase(PDS) play an important role in the physiological metabolism of carotene and virus induced silencing of foreign genes. The degenerated primers were designed from the conserved motifs of phytoene desaturase gene isolated from other plants in NCBI, and the middle fragments of PDS gene in Gossypium hirsutum were amplified. The 5' and 3' ends of the eDNA were cloned through RACE methods. The gene of PDS in Gossypium hirsutun was named as GhPDS1 (GenBank accession number: HQ441484). The semi-quantitative PCR results revealed that the transcript of GhPDS1 gene was the highest in flowers and then leaves. Phytoene desamrase gene is a endogenous gene as a visible marker to study the function of gene by endogenous gene silencing which causes cotton appear the yellow vein mosaic like pattern.%八氢番茄红素脱氢酶(PDS)在胡萝卜素生理代谢和病毒诱导外源基因沉默中起着非常重要的作用.本研究通过同源克隆方法,分析已报道植物八氢番茄红素脱氢酶氨基酸保守区,设计筒并引物,扩增出编码棉花PDS基因cDNA中间片段.通过RACE技术成功地克隆了棉花PDS基因的全长cDNA,命名为GhPDS1(GenBank No.HQ441184).利用半定量RT-PCR技术进行组织表达模式分析发现,该基因在花中表达量最高,其次为叶片.棉花PDS基因的成功克隆使得该基因可在病毒诱导的基因沉默研究中作为一个检测显示基因,它的沉默直接导致棉花叶片表现出斑驳、褪色等生物学特征,便于棉花中重要功能基因的深入研究.