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以两个耐贮性不同的青花菜高代自交系‘8554’和‘90196’为试验材料,分析其在贮藏过程中叶绿素含量的变化差异,利用实时荧光定量PCR技术对叶绿素降解相关基因的表达量进行研究。结果表明,在4℃贮藏期间,‘8554’和‘90196’的叶绿素含量均呈下降趋势,但耐贮藏材料‘8554’下降缓慢,且始终高于不耐贮材料‘90196’。Bo CLH1(叶绿素酶1)的表达量变化在上述两种材料中表现一致,均只在初始时检测到较高表达,后期表达量很低。Bo CLH2(叶绿素酶2)的表达量在‘8554’中变化较小,在‘90196’中变化较大,且高于‘8554’。Bo PPH(脱镁叶绿素酶)在‘8554’中一直呈下降趋势,而在‘90196’中贮藏14 d时有显著上升过程。Bo Pa O(脱镁叶绿酸a加氧酶)的表达量变化在二者中均有急剧增加的过程,但是在‘8554’中出现急剧增加的时期明显滞后于‘90196’。Bo RCCR(红色叶绿素代谢产物还原酶)在‘90196’中于贮藏21 d时出现一个明显的表达高峰,而在‘8554’中未出现。上述结果表明耐贮藏性不同的青花菜叶绿素降解机制不同,其衰老过程中叶绿素含量的变化主要通过Bo CLH2、Bo PPH、Bo Pa O以及Bo RCCR的表达量变化来调节,Bo CLH1主要在青花菜初始衰老中起降解叶绿素的作用。
Two different inbred lines of high-generation inbred lines ’8554’ and ’90196’ of broccoli were used as experimental materials to analyze their differences in chlorophyll content during storage. Real-time fluorescence quantitative PCR was used to analyze the correlation between chlorophyll degradation-related genes The amount of expression was studied. The results showed that the chlorophyll content of ’8554’ and ’90196’ showed a downward trend during storage at 4 ℃, but storage resistant material 8554 decreased slowly and remained consistently higher than that of ’90196’. The change of expression of Bo CLH1 (Chlase1) was consistent in the above two materials, and both the high expression was detected at the beginning and the low expression at the late stage. The expression level of Bo CLH2 (Chlase 2) changed little in ’8554’, changed greatly in ’90196’, and was higher than ’8554’. Bo PPH (pheophytinase) showed a downward trend in ’8554’, but there was a significant increase in ’90196’ when stored for 14 days. Bo Pa O (pheophorbide a oxygenase) showed a sharp increase in both, but the sharp increase in ’8554’ lagged to ’90196’. Bo RCCR (red chlorophyll metabolite reductase) showed a significant peak of expression in ’90196’ when stored for 21 days, but not in ’8554’. The above results indicated that the chlorophyll degradation mechanisms of different tolerant broccoli were different. The changes of chlorophyll content in the process of aging were mainly regulated by the expression changes of Bo CLH2, Bo PPH, Bo Pa O and Bo RCCR. Bo CLH1 was mainly found in broccoli Degradation of chlorophyll in initial senescence.