‘红肉蜜柚’果实汁胞粒化过程中POD同工酶谱分析  

Isonzyme analysis of POD from‘Hongroumiyou'fruit during the process of juicy sac granulation

作　　者：徐世荣[1,2] 潘鹤立 吴嘉玲[1,2] 丁安琪[1,2] 吕恃衡[1,2] 潘东明[1,2] 余磊[1,2] 张智伟[1,2] 李小婷[1,2] 

机构地区：[1]福建农林大学园艺产品贮运保鲜研究所,福州350002 [2]福建农林大学园艺学院,福州350002

出　　处：《果树学报》2016年第4期409-415,共7页Journal of Fruit Science

基　　金：福建省科技厅重大专项<亚热带名优水果育种技术开发和良种的示范推广>(2013NZ0002-1);国家科技支撑计划项目(2014BAD15B00);福建农林大学创新团队项目(cxtd12013)

摘　　要：【目的】探讨‘红肉蜜柚’果实汁胞粒化过程中过氧化物酶(POD)的同工酶变化及赤霉素对其的影响。【方法】以赤霉素处理和未经赤霉素处理的‘红肉蜜柚’果实的近中柱和远中柱汁胞为材料,统计粒化指数后,采用垂直板聚丙烯酰胺凝胶电泳方法进行可溶性和离子结合POD同工酶谱测定。【结果】发现‘红肉蜜柚’果实汁胞粒化指数随着果实生长发育呈先平稳后急剧上升的趋势,有无赤霉素处理无明显变化;可溶性POD中存在酸性POD和碱性POD,有无赤霉素处理对近、远中柱POD出现的时期和组分基本没有影响;离子结合POD只存在碱性POD,近中柱POD同工酶谱分离出6条(5条为果实发育后期的特异性谱带)、远中柱POD分离出4条,两者出现的时期和组分差异较大,赤霉素处理使近中柱5条果实发育后期特异性谱带减为3条且时期提前,而赤霉素处理下的远中柱没有检测到谱带。【结论】粒化时期,离子结合POD出现的5个特异性碱性POD可能与汁胞粒化关系密切,赤霉素处理可明显减少POD谱带,但赤霉素对汁胞粒化的影响有待进一步研究。【Objective】To test the variations of peroxidase（POD） isozymes during juice sac granulation in‘Hongroumiyou＇and to investigate the influence of gibberellin（GA） on this process.【Methods】‘Hon-groumiyou＇fruits were sampled from normal（95-211 d after flowering） and GA-treated（50 mg·L- 1）plants（127 d after flowering） and were used as the test plants in this study. Each group had 3 samples,and each sample was made up from a mixture of 3 fruits from different parts of the plant. Fruits wereweighed and their juice sac granulation index was recorded, and then the juice sacs near the column andaway from the column were sampled, treated with liquid nitrogen, and preserved at-80 ℃ for future use.Electrophoresis of the acidic POD isozymes： The buffer for the separation gel was 1.5 mol·L-1Tris-HCl,p H 8.8; the buffer for the stacking gel was 0.5 mol·L-1Tris-HCl, p H 6.8; and the stocking solution for thegel was 30%（m/V） acrylamide /bisacrylamide（29：1）. The buffer in the anode and cathode consisted of 200mmol·L-1glycine, 25 mmol·L-1Tris and 0.1% Tweens. The soluble and ion-bound crude enzyme solu-tions were respectively treated with 2× and 5× loading buffer（final concentration 50 mmol·L-1, with p H8.0 Tris-HCl, 10% glycerol and 0.01% bromophenol blue）, then mixed with Tweens（final concentration0.1%）, and centrifuged at 15 000 r·min-1at 4 ℃ for 10 min. The supernatant was then taken for loading.The soluble and ion-bound enzyme solutions were loaded at a volume of 10 μL and 40 μL respectively,and during electrophoresis, the stacking and separation voltages were 100 V and 150 V respectively. After electrophoresis, the gel was stained（100 m L staining solution with 0.1 g benzidine, 0.15 mol·L-1acetic acid, 0.15 mol·L- 1sodium acetate and 0.03% H2O2） and imaged. Electrophoresis of the alkaline POD isozymes： The buffer for the gel consisted of 0.6 mol·L-1HEPES, 1 mmol·L-1acetic acid and 10 mmol·L-1arginine; the stocking solution for the gel was 30%（m/V

关 键 词：'红肉蜜柚’ 粒化 POD 同工酶谱 

分 类 号：S666.3[农业科学—果树学]