附加秋水仙素培养基对烟草单倍体的加倍效应

Effect of medium added colchicine on chromosome doubling of tobacco haploid

【目的】探索秋水仙素在烟草单倍体染色体加倍应用中的新途径,并构建烟草DH(双单倍体)群体。【方法】以烟草‘TMK-12’ב吉烟5号’和‘NC71’ב净叶黄’2个杂交组合F1代花药为材料,在单倍体诱导培养基上进行单倍体诱导,待单倍体幼苗生长至1~2cm时,采用培养基附加秋水仙素加倍法,研究秋水仙素对烟草单倍体加倍效果的影响。【结果】 在单倍体诱导培养基上烟草花药发育产生大量的白色胚状体并进一步发育为单倍体苗。0.02%秋水仙素处理4d时,‘TMK-12’ב吉烟5号’和‘NC71’ב净叶黄’2个杂交组合烟草单倍体幼苗的存活率为66.56%~76.56%,染色体加倍率分别为50.60%和39.83%,利用这一技术体系建立了2个分别含61和68个加倍单倍体株的DH群体。【结论】烟草单倍体幼苗在附加0.02%秋水仙素的培养基中处理4d时加倍效果最佳,利用这一技术体系能够有效建立烟草DH群体。

【Objective】This study aimed to establish a new way to utilize colchicine in tobacco haploid chromosome doubling and construct DH(Doubled haploid)population.【Method】Anthers of F 1 generation of‘TMK-12’בJiyan 5’and‘NC71’בJingyehuang’were cultivated in the haploid induction medium.Then 1-2 cm haploid seedlings were transferred into colchicine medium to study the effect of colchicine on doubling efficiency of tobacco haploids.【Result】Tobacco anther produced a large number of white embryoid bodies and further developed into haploid seedlings on the haploid induction medium.When 0.02%colchicine was treated for 4 days,the survival rates of tobacco haploid seedlings of‘TMK-12’בJiyan 5’and‘NC71’בJingyehuang’were 66.56%-76.56%,and the chromosome doubling rates were 50.60%and 39.83%,respectively.Two DH populations containing 61 and 68 DH strains were successfully established using this technical system.【Conclusion】Haploid seedlings can be efficiently doubled by treating with 0.02%colchicine medium for 4 days.The tobacco DH population can be effectively established using this technical system.