Selection and characterization of ethionine-resistant alfalfa (Medicago sativa L.) cell lines
Selection and characterization of ethionine-resistant alfalfa (Medicago sativa L.) cell lines
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DOI：
10.1007/bf00285896
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发表时间：
1981-03
影响因子：
5.4
通讯作者：
B. Reisch;S. H. Duke;E. Bingham
中科院分区：
文献类型：
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作者：
B. Reisch;S. H. Duke;E. Bingham
以具有组织培养再生能力的二倍体苜蓿（HG 2）为材料，筛选出抗蛋氨酸类似物乙烯利生长抑制的变异细胞系。用甲磺酸乙酯对约107个悬浮培养细胞进行诱变，然后将其接种在含有乙醇的固体培养基中。在含有0.02 mM乙醇的培养基上形成愈伤组织菌落。在回收的124个细胞系中，有91个再生植株。在不含乙醇的培养基上生长6个月后，110个愈伤组织细胞系中有15个细胞系在1 mM乙醇上的生长显著优于HG 2。几种抗乙硫氨酸的愈伤组织培养物也对生长抑制具有抗性，这是由于向培养基中添加赖氨酸+苏氨酸。高浓度，相对于HMG2愈伤组织，蛋氨酸，半胱氨酸，胱硫醚，谷胱甘肽被发现在一些，但不是所有的，抗乙硫氨酸愈伤组织培养。细胞系R32的可溶性甲硫氨酸增加了约10倍，与HMG2愈伤组织相比，总游离氨基酸增加了43%，蛋白质中氨基酸增加了40%。蛋白质中每种氨基酸的相对量在两者中是相同的。
Diploid alfalfa (HG2), capable of plant regeneration from tissue culture, was used to select variant cell lines resistant to growth inhibition due to ethionine (an analog of methionine). Approximately 107suspension-cultured cells were mutagenized with methane sulfonic acid ethylester and then plated in solid media containing ethionine. Callus colonies formed on media with 0.02 mM ethionine. Of the 124 cell lines recovered, 91 regenerated plants. After six months growth on media without ethionine, 15 of 110 cell lines of callus grew significantly better than HG2 on 1 mM ethionine. Several ethionine-resistant callus cultures were also resistant to growth inhibition due to the addition of lysine + threonine to the media. High concentrations, relative to unselected HG2 callus, of methionine, cysteine, cystathionine, and glutathione were found in some, but not all, ethionine-resistant callus cultures. Cell line R32, which had aca.tenfold increase in soluble methionine, had a 43% increase in total free amino acids and a 40% increase in amino acids in protein as compared to unselected HG2 callus. Relative amounts of each amino acid in protein were the same in both.