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作 者 ：张晓东，林廷安

期 刊 ：核农学报 1994年 8卷 01期 页码：0-0

关键词：细胞；悬浮培养；辐射；PEG；苜蓿；

Keywords:cell suspension culture, irradiation, PEG, alfalfa,

摘 要 ：本文对紫花苜蓿的细胞悬浮培养、悬浮细胞系的建立、离体辐射效应和高浓度ＰＥＧ耐受性变异体的筛透进行了研究。结果表明：１．高浓度的２，４．Ｄ（５～１０ｍｇ／Ｌ），低浓度的ＡＢＡ（０．１～２ｍｇ／Ｌ），高浓度的蔗糖（６％～９％），较高的离子强度，以及逐步缩短继代时间，均有利于胚性悬浮细胞系的快速建立。相反，细胞分裂素，高浓度的ＮＨ和ＧＡａ的存在，不利于悬浮系的建立。细胞分裂素（４ＰＵ３０，ＫＴ或ＢＡ）可促进细胞的团聚，不利于细胞的分散。此外，低浓度的２，４－Ｄ，４ＰＵ３０（或ＢＡ，ＫＴ）和ＡＢＡ的配合使用，在悬浮培养基中可形成大量的体细胞胚。２．辐射处理悬浮细胞的最适剂量为２０～６０Ｇｙ。３．未经改造的初期悬浮培养物远较悬浮细胞系对高水势敏感。４．胚性悬浮细胞系经２０Ｇｙ的γ射线处理，筛选出了对１５％ＰＥＧ水势（约－１１巴）耐受性的克隆６个，并获得了一批再生植株，从２０％ＰＥＧ（约－１５巴）筛选出１个抗性克隆。抗性细胞系对高浓度ＰＥＧ和ＮａＣｌ产生的水势在细胞水平表现出很强的抗性。

Abstract:Elements affecting suspension cell culture in alfalfa(Medicago sativa L)were studiedand a method of rapid establishment of embryogenlc suspension cell lines was introduced.Effccts ofγ ray irradiation on the growth of suspension cells were studied,and the optiumdose of irradlation for inducing mutants from suspcnsion cells was about 20～60Gy.Effectsof PEG and NaCl induced water stress on the growth of suspension cells were also investigat-ed,and the results showed that the congregants of preliminary suspension culture were moresusceptible than the established suspension cell lines.With 20Gy of γ ray irradiation on sus-pension cell line(JL416.six clones were obtained with 70days of selection on medium of15% PEG(about-11bar).A number of regenerated plants were obtained from these clones,One clone was also gained from medium containing 20%PEG(about-15bar).The selectedmutant cell lines(JP15 and JP20)had strong resistances to high concentration of PEG andNaCl induced water stress.

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