基于实时荧光定量PCR技术的蝴蝶兰PPO和PAL基因表达分析  

Phalaenopsis aphrodite Rchb.F.PPO and PAL Expression Based on Real-time PCR Technology

作　　者：赵和文[1] 崔金腾[1] 周田田[1] 

机构地区：[1]北京农学院园林学院,北京102206

出　　处：《中国农学通报》2015年第7期125-130,共6页Chinese Agricultural Science Bulletin

基　　金：北京市属高等学校创新团队建设与教师职业发展计划项目(IDHT20150503)~~

摘　　要：为研究蝴蝶兰在组织培养过程中的褐化机制,通过观察与褐化相关基因PPO和PAL在组织培养过程中不同时期的表达量,判断上述2个基因对褐化的影响。采用实时荧光定量的方法,设计特异性引物,获得PPO基因与PAL基因的目的片段,再以实时荧光定量方式,检验上述2个基因在蝴蝶兰组培苗中的表达量。结果表明,获得的PPO基因目的片段与蝴蝶兰×五唇兰相似性达到97%,蝴蝶兰PPO基因目的片段序列与蝴蝶兰×朵丽兰相似性达到98%。通过对蝴蝶兰组培苗培养不同阶段中PPO基因及PAL基因的表达差异研究,发现：在对组培苗叶片切割后PPO基因的表达量迅速增加,随培养时间的推移,表达量经过缓慢的降低后逐渐增高,在10~12天阶段表达量急剧增加,在12天达到最大量,随后降低,后期表达量浮动不大;PAL基因分别在1、8、12天表达量急剧增加,12天的表达量最大,随后,总体表达量有降低趋势。2个基因在第12天表达量都达到最高,说明此时是褐化发生的关键时期,若此时期对其进行有效预防措施,将会减少褐化现象。The paper aims at researching browning of tissue cultured-seedling of Phalaenopsis aphrodite Rchb.F., in order to judge the influence of PPO gene and PAL gene by observing their expression at different tissueculture time. We obtained the target sequences of PPO gene and PAL gene by designing specific primers andtested the expression of the above mentioned genes by real-time PCR. In our study, the similarities between Phalaenopsis aphrodite Rchb. F. PPO and Phalaenopsis aphrodite Rchb. F.×Pulcherrima PPO were up to 97%and the similarities between Phalaenopsis aphrodite Rchb. F. PPO and Phalaenopsis aphrodite Rchb. F.t ×Doritis PAL were up to 98%. We found that after slicing the leave of tissue cultured-seedling, the expression of PPO dramatically increased, and as time went on, the expression first slowly decreased and then increased.The expression dramatically increased between 10 days and 12 days and then was up to the maximum;following that, the expression reduced and did not change much. The expression of PAL gene rapidly increased,respectively at 1, 8 and 12 days, and the expression was the maximum at 12 days. At last, the expression wasdown. Both PPO and PAL expression reached the maximum at 12 days, which indicated that it was the criticalperiod for browning. Precaution taken at that time could reduce browning.

关 键 词：组织培养 褐化 PPO基因 PAL基因 

分 类 号：S687.3[农业科学—观赏园艺]