以陆地棉种质系珂字312和陆地棉标准系TM-1为材料，结合优化的陆地棉体细胞培养体系，系统地研究了琼脂、Gelrite和Phytagel 3种培养基凝固剂对愈伤组织的诱导、胚性愈伤的增殖、体细胞胚状体的发生和植株再生的影响。结果表明，用Phytagel作为培养基的凝固剂，可显著改善陆地棉体细胞培养过程中的褐化问题，并具有提高胚性愈伤组织增殖速度、体细胞胚胎发生和植株再生频率等作用。Gelrite虽也能改善棉花体细胞培养中的褐化问题，但因水化现象严重而影响胚性愈伤组织增殖和体细胞胚状体发生，其效果不如Phytagel。因此，在陆地棉体细胞培养过程中的胚性愈伤组织诱导、体细胞胚状体发生和植株再生过程中，宜用Phytagel作为培养基的凝固剂，而无菌苗培养和愈伤组织诱导仍可以选用琼脂凝固剂。
Abstract
The effects of the three different gelling agents, agar, gelrite and Phytagel, on callus inducing, embryogenic callus amplification, the somatic embryogenesis and plant regeneration of upland cotton (Gossypium hirsutum L.) germplasms, Coker 312 and TM-1, were studied systematically, using the system of upland cotton somatic culture developed in our laboratory. The results showed that the phytagel which used as the gelling agent for the medium of somatic culture could improve the browning problem during the somatic culture, increase the amplification of embryogenic callus, and improve the quality of the callus and embryogenic callus, which led to significantly increasing in somatic embryogenesis and plant regeneration. So the phytagel is a good gelling agent for medium of somatic culture in upland cotton. Although the gelrite also can solve the problem of the browning during the somatic culture, due to its hydrating process during the culture which may affect the amplification of embryogenic callus and somatic embryogenesis seriously, the results of gelrite is not as good as phytagel. Combination with the method of somatic culture system optimized by our laboratory recently, the regeneration plants from 10 upland cotton genotypes, included the genotypes called the recalcitrant nature in plant regeneration, were obtained in a relative short period of culture, using the Phytagel as the gelling agent. Therefore, it is suggested that Phytagel should be used as the gelling agent in the media for inducing embryogenic callus, somatic embryogenesis, and plant regeneration during the somatic culture of upland cotton, especially for the genotypes with recalcitrant nature in plant regeneration. While for the culture of germ-free seedlings and callus inducing, the agar is still a good gelling agent because of the economic reason.
关键词
陆地棉
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体细胞培养 /
凝固剂 /
植株再生
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Key words
upland cotton
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somatic culture /
gelling agent /
plant regeneration
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