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为了检测上饶早梨离体保存库中不同再生苗的遗传稳定性,采用聚丙烯酰胺凝胶电泳技术对其20个样本进行多酚氧化酶(PPO)同工酶、超氧化物歧化酶(SOD)同工酶、淀粉酶同工酶、过氧化物酶(POD)同工酶、酯酶(EST)同工酶、过氧化氢酶(CAT)同工酶和蛋白酶同工酶分析。结果表明,20个样品的POD同工酶、EST同工酶和蛋白酶同工酶均只有一条带;20个样品的CAT同工酶均出现两条带;PPO同工酶在20个样品中出现6条带;SOD同工酶在20个样品中出现5条带;淀粉酶同工酶在20个样品中出现4条带。利用popgene软件计算出POD同工酶、EST同工酶、蛋白酶同工酶、CAT同工酶、PPO同工酶、SOD同工酶和淀粉酶同工酶的多态性百分率分别为0%、0%、0%、0%、83.33%、60%和100%。使用NTsys 2.1软件计算PPO同工酶、SOD同工酶和淀粉酶同工酶酶带的遗传相似系数,并用非加权组平均法(UPGMA)进行聚类分析。结果表明,3个同工酶的聚类结果一致,20个样本分为两大组,一组包括“花厅六月雪”和“花厅黄皮消”两个主栽品种,另一组只包括“田墩六月雪”一个主栽品种。3个主栽品种内的样本所有同工酶酶带结果一致且遗传相似系数均为1.000 0,说明3个主栽品种内样本的遗传稳定性高,无遗传变异。本试验结果表明,上饶早梨离体保存库的建立具有较强的可靠性和可行性。
In order to detect the genetic stability of different regeneration seedlings in vitro in Shangrao early pear, the polyphenol oxidase (PPO) isoenzyme, superoxide dismutase (SOD) ) Isozyme, amylase isozyme, peroxidase (POD) isozyme, esterase (EST) isozymes, catalase (CAT) isoenzyme and protease isoenzyme analysis. The results showed that there were only one band of POD isozyme, EST isozyme and protease isozyme in 20 samples, two bands of CAT isozyme in 20 samples, and PPO isozymes appeared in 20 samples 6 bands; SOD isozyme in 20 samples appeared in 5 bands; amylase isozyme in 20 samples appeared in 4 bands. The percentages of POD isozymes, EST isozymes, protease isozymes, CAT isozymes, PPO isozymes, SOD isozymes and amylase isozymes were calculated using popgene software. The percentages of polymorphisms were 0% 0%, 0%, 0%, 83.33%, 60% and 100%. The genetic similarity coefficients of PPO isozymes, SOD isozymes and amylase isoenzyme bands were calculated using NTsys 2.1 software, and cluster analysis was performed by using unweighted group average method (UPGMA). The results showed that the three isozyme clustering results were consistent, 20 samples were divided into two groups, a group, including “Flower Hall June Snow” and “Flower Hall Huangpi Xiao” two major cultivars, and the other One group includes only one major cultivar “Tian Jun June Snow”. The results of all isoenzyme bands in three main cultivars were consistent and the genetic similarity coefficients were 1.000 0, indicating that the genetic stability of the three main cultivars was high and there was no genetic variation. The results of this experiment show that the establishment of the ex vivo preserved bank of Shangrao early pear has strong reliability and feasibility.