红叶芥低温胁迫下苯丙氨酸解氨酶活性及其基因的克隆表达
Cloning and Expression of PAL Gene and PAL Activity Assay in Red-Leaf Mustard(Brassica juncea var.garrhiza Tsen et Lee)under Low Temperature Stress
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摘要: 用8℃低温处理红叶芥四叶期植株,在低温处理的52 h内分别取样6次,取样后分别提取叶片总RNA,再利用RT-PCR克隆得到红叶芥PAL基因开放阅读框内1908 bp片段,核苷酸序列分析显示,红叶芥PAL基因与芜青PAL基因的同源性达到99，.半定量RT-PCR结果显示,在8℃低温胁迫下,红叶芥PAL基因表达量呈现先降低后升高的趋势,在处理18 h后达到最低点,随后迅速升高,这与PAL活性变化一致.Abstract: The 4-leaf seedlings of red-leaf mustard(Brassica juncea var.garrhiza Tsen et Lee)were exposed to low temperature(8 ℃)for 52 h,during which period leaf samples were taken 6 times for total RNA extraction.The open reading frame fragment that was generated by RT-PCR using specific primers was 1 908 bp.The cDNA from the leaves of red-leaf mustard was used as the template.Nucleotide sequence analysis showed that red-leaf mustard PAL gene shared 99% identity to that of Brassica napus.In response to the low-temperature stress in this experiment,the expression of PAL first dropped and then increased rapidly,being the lowest 18 h after the start of the treatment,a similar trend to that of PAL activity.
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