秋水仙碱处理诱导银灰杨2n花粉与杂种三倍体创制  

Pollen chromosome doubling induced by colchicine treatment and creation of hybrid triploids in Populus canescens

作　　者：周晴 吴剑 桑亚茹 赵征洋 刘美芹[1] 张平冬[1] Zhou Qing;Wu Jian;Sang Yaru;Zhao Zhengyang;Liu Meiqin;Zhang Pingdong(National Engineering Laboratory for Forest Tree Breeding,School of Biological Sciences and Biotechnology,Beijing Forestry University,Beijing 100083,China)

机构地区：[1]林木育种国家工程实验室北京林业大学生物科学与技术学院,北京100083

出　　处：《北京林业大学学报》2020年第3期119-126,共8页Journal of Beijing Forestry University

基　　金：国家自然科学基金项目(31570646)。

摘　　要：【目的】为创制银灰杨杂种三倍体新种质,研究完善银灰杨三倍体育种技术体系,筛选生长迅速、抗逆性强的银灰杨新品种。【方法】本研究以银灰杨、河北杨以及银腺杨为试验材料,在优化秋水仙碱诱导银灰杨花粉染色体加倍技术条件的基础上,开展了秋水仙碱注射处理对2n花粉粒表面结构的影响以及杂交创制杂种三倍体的研究。【结果】减数分裂时期、注射次数、减数分裂时期与注射次数的交互效应均对2n花粉诱导率具有极显著影响。利用0.5%的秋水仙碱溶液对发育至粗线期的花粉母细胞进行11次注射处理是秋水仙碱诱导银灰杨花粉染色体加倍的最佳处理组合,2n花粉平均诱导率最高可达(30.27±8.69)%。与天然2n花粉粒相比,秋水仙碱诱导型2n花粉粒表面结构出现了明显的裂沟,说明秋水仙碱注射处理对2n花粉花粉壁形成具有一定的影响。选取含有一定2n花粉比例的花粉,分别与河北杨、银腺杨雌配子进行授粉杂交,共收获杂种苗木4 955株,检测出三倍体5株,其中,2株三倍体来源于河北杨×银灰杨的杂交组合;另3株三倍体来源于银腺杨×银灰杨的杂交组合。【结论】秋水仙碱能高效诱导银灰杨花粉染色体加倍并获得2n花粉,但是注射会影响2n花粉花粉壁的形成,使得2n花粉粒表面结构产生裂沟。秋水仙碱诱导的2n花粉可以用于授粉杂交,成功获得杂种多倍体植株。[Objective] Improving the technical system of triploid breeding to generate new triploid germplasm can serve for developing new varieties with fast growing and higher resistance in P. canescens.[Method] Based on the optimization of technical conditions for inducing pollen chromosome doubling,effects of colchicine on induced 2 n pollen of surface structure and triploid production by crossing induced 2 n pollen were conducted by taking P. canescens, P. hopeiensis and P. alba × P. glandulosa as experimental materials. [Result] The results showed that meiotic stages, injection times and the interaction between meiotic stage and injection time had highly significant effects on frequency of colchicine-induced 2 n pollen.The optimum treatment combination to induce pollen chromosome doubling was to inject 11 times with0.5% colchicine solution when pollen mother cells were at pachytene. The highest mean frequency of colchicine-induced 2 n pollen was(30.27 ± 8.69)%. Compared with natural 2 n pollen, some narrow furrows on the surface of the colchicine-indued 2 n pollen were observed, suggesting that colchicine did, to some extent, affect the formation of cell wall of induced 2 n pollen. 4 955 hybrid offsprings were obtained by crossing induced 2 n pollen with female gametes of P. hopeiensis and P. alba × P. glandulosa, and five real triploids were detected by flow cytometric analysis and somatic chromosome counting. Among the five triploids, two triploids were derived from the cross combination between P. hopeiensis and P. canescens,and three triploids were derived from the cross combination between P. alba × P. glandulosa and P.canescens. [Conclusion] Colchicine could not only induce pollen chromosome doubling to produce 2 n pollen but also affect the formation of cell wall of colchicine-induced 2 n pollen and resulted in narrow furrows on ectexine structure in P. canescens. Hybrid polyploid offsprings were successfully obtained by crossing colchicine-induced 2 n pollen.

关 键 词：银灰杨 秋水仙碱 2n花粉诱导 花粉粒表面结构 三倍体 

分 类 号：S722.35[农业科学—林木遗传育种] S792.11[农业科学—林学]