白花除虫菊四倍体与二倍体株系抗氧化酶活性研究（摘要）

[目的]测定白花除虫菊四倍体株系与二倍体对照株系试管苗和田间叶片中SOD、POD及APX酶的活性,为选育抗性强的白花除虫菊优良四倍体品种提供理论依据。[方法]先加入预冷的提取缓冲液（pH=7.5,0.05 mol/L Tris-Hcl缓冲液）冷冻离心提取叶片中的各酶液。蛋白含量和SOD活性的测定按南京建成生物工程研究所试剂盒说明书操作。POD总活力测定：100μl酶液用Tris-HCl（pH=7.0）溶液稀释至1 ml,准确加入0.1%的愈创木酚1 ml,30℃水浴10 min后,立即测定溶液在470 nm处的吸光度,然后准确加入0.08%的H_2O_2 100μl,充分混匀,反应2 min时立即测定溶液在470 nm处的吸光度,求出反应前后的吸光度差值。酶的比活力用ΔOD_（470）/（mg·min）表示。APX活性测定：3 ml反应液中含50.0mmol/L PBS（pH=7.0）,0.1 mmol/L EDTA,0.1 mmol/LH2O2,0.5 mmol/LASA,最后加入100μl酶液。在室温下记录10 s,30 s时290 nm处吸光度的变化。酶活单位定义为每小时氧化1μmol ASA的酶量为一个酶活单位。[结果]四倍体株系与二倍体对照株系比较,其保护酶系统中各种抗氧化酶的活性都能得到普遍提高,并且各株系的田间样品与试管苗样品的各抗氧化酶活性呈正相关,因此可以应用组织培养技术在育种的初期试管苗阶段进行早期筛选,缩短选育时间,节约人力物力。[结论]试管苗期间各株系的抗氧化酶活性可以作为筛选抗病抗逆性强的白花除虫菊优良株系的参考指标。

Antioxidase Activities of Tetraploid Lines and Diploid Control Line of Chrysanthemum cinerariifolium

Objective The SOD, POD and APX enzyme activities of tetraploid lines and diploid control line of Chrysanthemum cinerariifolium in test-tube and field were determined in order to provide a reference for the selective breeding of high resistance good tetraploid varieties of C. cinerariifolium. Method The SOD, POD and APX activities of each sample were determined. Every material should have 3 replicates which been determined and calculated mean value to analyze. Result The various kinds of antioxidase activities in protective enzyme system of tetraploid lines were increased generally in comparison with that of diploid control line. And there is certain relativity between the samples in field and in test-tube of these lines. Conclusion The antioxidase activities of each line in test-tube could be considered as reference index to screen high resistance good tetraploid varieties of C. cinerariifolium.