利用cDNA-AFLP技术分析旱作促进水稻种子根伸长过程中的基因表达

摘要

The seminal root elongation of rice under short-duration upland condition was stimulated. cDNA amplified fragment length polymorphism (cDNA-AFLP) analysis was introduced to identify differentially expressed genes in seminal root tips under upland conditions, and to compare spatial expression in three root tissues-seminal root tips, lateral roots and adventitious root priomordia exposed to upland condition. Of the 1 640 fragments inspected, 70 showed an up-regulated expression pattern in seminal root tips under upland conditions. Twenty-four fragments were cloned and sequenced. Two fragments showed significant similarity to PDK and APRT. A full-length cDNA of rice APRT was obtained by electronic splicing. One cDNA extended by RACE showed no significant similarity to known sequences in the database. Northern blot analysis confirmed the expression of these three genes. This is the first report of the use of cDNA-AFLP to study differential gene expression in rice roots.%短期旱作促进水稻种子根的伸长.利用cDNA-AFLP技术分析种子根根尖在旱作条件下差异表达的基因,同时比较这些基因在种子根尖、侧根和不定根原基区的表达差异.在1 640个片段中,70个在种子根根尖中受旱作诱导,其中24个被克隆并测序.2个基因分别编码丙酮酸脱氢酶激酶(PDK)和腺嘌呤转磷酸核糖基酶(APRT),并用电子拼接技术获得水稻的APRT全长cDNA;另一个经cDNA末端快速扩增法延长后仍无同源序列.Northern杂交验证了这3个基因的cDNA-AFLP表达谱.这是首次报告使用cDNA-AFLP技术研究水稻根组织的差异表达基因.