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结球甘蓝小孢子胚胎发生的细胞学研究

DOI: 10.7685/j.issn.1000-2030.2014.05.008, PP. 47-54

Keywords: 胚胎发生,细胞程序性死亡,发育途径,结球甘蓝

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Abstract:

游离小孢子培养即小孢子在外界胁迫诱导的条件下,从其配子体发育途径转向孢子体发育途径,诱导小孢子胚胎发生的过程。本研究通过对结球甘蓝小孢子胚胎发生的细胞学观察,为甘蓝游离小孢子培养中的可调控性及相关胚胎发育研究提供理论依据。选用3个在游离小孢子培养中胚胎发生响应有差异的甘蓝杂交种,分别采用醋酸洋红、4’,6-二脒基-2-苯基吲哚(DAPI)及荧光素双醋酸酯(FDA)3种染色方法,对甘蓝小孢子体内配子体发育、游离小孢子培养中的胚胎发生过程及细胞死亡现象进行了观察。结果表明小孢子正常发育方向是单核小孢子经历2次不对称核分裂后发育成三核花粉,而单核靠边期小孢子在体外经32.5℃热激胁迫1d后,小孢子绝大部分死亡,培养3d后小孢子死亡率达到90%左右。结球甘蓝游离小孢子培养胚胎发生途径以B途径为主第1次分裂为对称分裂,其后2个核进行反复分裂形成多细胞团,依次经历球型胚、心型胚、鱼雷胚,直至子叶胚的形成。除了以上典型B途径外,培养过程中还同时存在多种发育途径第1次不对称分裂后,生殖核继续分裂即为A-G途径;营养核和生殖核两者同时继续分裂的A-VG途径。结论甘蓝小孢子在体外经32.5℃热激胁迫1d后,B途径为其主要孢子体发育途径;热激胁迫迅速诱导了小孢子的死亡。

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