摘要
为了快速、准确地统计甘薯小孢子活力,确定合适的培养方式,以甘薯栽培品种“冀紫3号”单核期小孢子为试验材料,从工作液浓度、染色温度和染色时间3个方面,对FDA荧光染色法进行优化;并采用优化后的方法,对不同减数分裂时期和采用不同液体培养基培养的小孢子进行了活力检测。结果表明:常温(25℃)下FDA工作液浓度8μg/mL、染色5min,可以简便、有效地检测甘薯单核期小孢子的活力。该方法还可应用于小孢子母细胞时期小孢子活力的检测,但是不适用于甘薯小孢子四分体时期和成熟花粉粒期的小孢子活力测定。培养过程中4种液体培养基培养的小孢子活力变化趋势基本一致,均表现为培养开始后活力快速降低,第2天时小孢子活力仅为培养开始时的50%左右,第10天时小孢子活力趋于0。研究结果不仅可为甘薯小孢子活力测定提供依据,还可为甘薯单倍体育种提供一定的参考。
In order to quickly and accurately calculate the vitality of sweet potato microspores and determine the appropriate cultivation method,the FDA fluorescence staining method was optimized from three aspects:working solution concentration,staining temperature,and staining time,using the mononuclear microspores of sweet potato cultivar Jizi 3 as the experimental material.And the optimized methods were used to detect the vitality of microspores cultured in different liquid media at different stages of meiosis.The results showed that at room temperature(25℃),FDA working solution concentration of 8μg/mL and staining for 5 minutes can easily and effectively detect the vitality of sweet potato mononuclear microspores.This method can also be applied to the detection of microspore vitality during the microspore mother cell stage,but it is not suitable for the determination of microspore vitality during the sweet potato microspore tetrad stage and mature pollen grain stage.During the cultivation process,the trend of microspore vitality changes in the four liquid culture media was basically consistent,showing a rapid decrease in vitality after the start of cultivation.On the second day,microspore vitality was only about 50%of that at the beginning of cultivation,and on the tenth day,microspore vitality tended to O.The research results can not only provide a basis for the determination of microspore vitality in sweet potato,but also provide some reference for haploid breeding of sweet potato.
作者
陈敬辉
高润红
焦伟静
王颖帅
胡亚亚
陈典典
刘兰服
马志民
刘艳芬
CHEN Jing-hui;GAO Run-hong;JIAO Wei-jing;WANG Ying-shuai;HU Ya-ya;CHEN Dian-dian;LIU Lan-fu;MA Zhi-min;LIU Yan-fen(Hebei University of Engineering,Handan 056038,China;Institute of Grain and Oil Crops,Hebei Academy of Agriculture and Forestry Sciences,Shijiazhuang 050035,China;Institute of Biotechnology,Shanghai Academy of Agricultural Sciences,Shanghai 201106,China;Hengshui University,Hengshui 053000,China)
出处
《河北农业科学》
2025年第2期102-108,共7页
Journal of Hebei Agricultural Sciences
基金
河北省农林科学院科技创新专项课题项目(2022KJCXZX-LYS-12)
河北省现代农业产业技术体系创新团队项目(HBCT2023060202)
国家甘薯产业技术体系项目(CARS-10)
河北省重点研发计划项目(22322911D)。
关键词
甘薯
FDA荧光染色
小孢子活力
单核期
Sweet potato
FDA fluorescent staining
Microspore viability
Mononuclear phase
作者简介
陈敬辉(2000-),男,河北曲阳人,硕士研究生在读,研究方向为作物遗传育种。E-mail:2090840575@qq.com;通信作者:刘艳芬(1968-),女,河北徐水人,教授,硕士,主要从事植物组织培养与栽培应用研究。E-mail:274821817@qq.com。
