甘蓝小孢子单倍体植株加倍技术探讨  

Technology discussion of doubling cabbage haploid plants

作　　者：程芳芳[1] 张恩慧[1] 杨安平[2] 程永安[1] 许忠民[1] 董韩[1] 霍柳青 

机构地区：[1]西北农林科技大学园艺学院,陕西杨凌712100 [2]杨凌职业技术学院生物工程系,陕西杨凌712100

出　　处：《西北农林科技大学学报（自然科学版）》2015年第6期167-173,共7页Journal of Northwest A&F University(Natural Science Edition)

基　　金：国家科技支撑计划项目(2012BAD02B01);西北农林科技大学农业科技推广基金项目(NYY2012-10);陕西省科学技术研究发展计划项目(2012NKC02-05;2012KTCL02-09);杨凌职业技术学院科学研究基金项目(A2010014);陕西省教育厅科学研究项目(自然科学专项)(11JK0642)

摘　　要：【目的】比较甘蓝单倍体植株(H植株)茎生叶及其愈伤组织的秋碱处理时间及愈伤组织大小对再生植株加倍效果的影响,筛选加倍效果最佳的秋碱处理时间与合适的外植体,为扩大甘蓝小孢子双单倍体植株(DH株)群体及提高甘蓝小孢子培养效率奠定基础。【方法】以3个甘蓝H植株(H10-23,H10-3和H10-5)的茎生叶为试材,用2.0mg/L秋碱溶液浸泡处理H植株叶片外植体及其愈伤组织以获得再生DH植株,设置15,30和60h3个处理时间,比较分析秋碱处理叶片外植体和愈伤组织的时间对加倍效果的影响;同时用2.0mg/L秋碱溶液处理大(直径≥1cm)、中(1cm>直径≥0.5cm)、小(直径<0.5cm)3种类型的愈伤组织30h,比较愈伤组织大小对加倍效果的影响。【结果】2.0mg/L秋碱处理甘蓝单倍体植株叶片外植体30h,可获得44.80%的平均再生DH株加倍率;2.0mg/L秋碱处理愈伤组织30h,可获得58.13%的平均再生DH株加倍率;2.0mg/L秋碱处理中等大小的愈伤组织30h,可获得51.11%的再生DH株加倍率。【结论】用2.0mg/L秋碱处理甘蓝H植株茎生叶及其愈伤组织30h,可获得较高比例的DH株。【Objective】This study investigated the effect of colchicine treatment time of cauline leaves in cabbage haploid plants(H plants)and size of callus in colchicine on doubling effect of regenerated plants to screen the optimal treatment time and the suitable explants for expanding cabbage double haploid plants(DH plants)groups and improving culturing efficiency of cabbage microspore.【Method】Using cauline leaves of three H plants(H10-23,H10-3and H10-5)as test materials,2.0mg/L colchicines was used to treat leaf explants of H plants and callus to obtain regenerated DH plants.Three treatment time periods(15h,30 hand 60h)and 3types of callus(large callus with diameter of larger than 1cm,medium with diameter of larger than 0.5cm and less than 1cm,and small with diameter of less than 0.5cm)with treatment time of 30 hwere tested.【Result】Average double rates of DH plants by treating the explant of cabbage leaf blade,callus and medium callus in 2.0 mg/L colchicine for 30 h were 44.80%,58.13% and51.11%,respectively.【Conclusion】Cauline leaf of cabbage H plants and its callus soaked in 2.0mg/L colchicine for 30 hhad high DH plant ratio.

关 键 词：秋水仙素 DH株 愈伤组织 单倍体 甘蓝 叶片再生 加倍技术 

分 类 号：S[农业科学]