油菜花粉超低温保存及花粉原生质体分离

油菜花粉超低温保存及花粉原生质体分离 POLLEN CRYOPRESERVATION AND POLLEN PROTOPLAST ISOLATION IN BRASSICA NAPUS 梁立 Liang Li Xu Bingfang Zheng Congyi Zhou Chang 1 2 first-author 徐秉芳 1 2 郑从义 1 2 周嫦 1 2 武汉大学生命科学学院 武汉大学生命科学学院 School of Life Science,Wuhan University School of Life Science,Wuhan University 武汉大学中国典型培养物保藏中心 武汉大学中国典型培养物保藏中心 <正> 花粉超低温保存有多方面的研究与应用价值,并已在多种植物的成熟花粉保存方面取得长足进展.幼嫩花粉保存方面开始有个别报道.近年在花粉原生质体的实验研究中,为了减少对开花季节的依赖,我们作了紫菜苔不同发育时期花粉的超低温保存及其花粉原生 A cryopreservation procedure of young （late uninucleate to early binucleate）and mature pollen in Brassica napus L.with high survival was established.Pollen grains suspended in the cryoproteetant （15% sucrose,10 % DMSO in B 5 medium） were frozen by a three-step method :frozen atarateof 1℃·min -1 from 0℃·min -1 to-10℃,hold for 15 min,then by 1℃·min -1 to -40℃,hold for 1 h,and transfered to liquid nitrogen and later thawed in water bath at 40℃.The relative survival rates were 94.% and 97.9% in the case of young and mature pollen respectively. Both preserved and fresh young pollen could be triggered to cell divisions leading to multicellular clusters.Mature pollen protoplasts could be isolated successfully from fresh as well as preserved pollen with frequencies of 78.1% and 76.9% respectively.For isolation of young pollen protoplasts anineubation of pollen in NLN medium at 35℃ before maceration was needed giving rise the isolation rates of 34.7% and 18.8% for fresh and preserved materials respectively. Q944.42 国家自然科学基金资助的课题 1993-06-01 2021-04-01 6