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为了提高体细胞耐盐突变体筛选的效率,本试验研究不同浓度氯化钠(NaCl)对百喜草愈伤组织分化和植株再生的影响。在琼脂浓度为8 g/L和16 g/L的继代培养基上,以百喜草品种太阳花的成熟种子分别诱导产生湿润、粘连和干燥、松散2种类型的颗粒状愈伤组织。在NaCl浓度为0(对照)、1.2%、1.6%、2.0%的培养基上分别筛选30 d,干燥、松散颗粒状愈伤组织的绿芽分化率分别为36.1%、20.4%、19.0%和9.3%,绿芽的植株再生率分别为52.6%、46.3%、30.0%和20.6%;湿润、粘连颗粒状愈伤组织的绿芽分化率分别为15.3%、3.6%、0.7%和0,绿芽的植株再生率分别为25.8%(对照)、16.7%(1.2%NaCl)和0(1.6%NaCl)。表明干燥、松散的颗粒状胚性愈伤组织适宜被选做体细胞耐盐突变体筛选材料。SSR分子标记检测结果显示,479对引物组合中有8对引物在百喜草田间实生苗、组织培养再生植株和不同NaCl浓度(1.2%、1.6%、2.0%)筛选得到的再生植株混合株之间PCR扩增存在多态,其中2对引物WXE-30和WXE-246适用于耐盐突变体分子鉴定。
In order to improve the screening efficiency of somatic salt tolerant mutants, we studied the effects of different concentrations of sodium chloride (NaCl) on the callus differentiation and plant regeneration of hippophae rhamnoides. Two types of granule callus, wet, adhesive, dry and loose, respectively, were induced in the mature medium of the hippocampus Blossom on the subculture medium with agar concentrations of 8 g / L and 16 g / L. After 30 days of NaCl concentration 0 (control), 1.2%, 1.6% and 2.0%, respectively, the rate of green shoot differentiation of dry and loose granular callus was 36.1%, 20.4% and 19.0%, respectively 9.3%, and green shoot regeneration rates were 52.6%, 46.3%, 30.0% and 20.6%, respectively. The rate of green shoots budding was 15.3%, 3.6%, 0.7% and 0% Plant regeneration rates of shoots were 25.8% (control), 16.7% (1.2% NaCl) and 0 (1.6% NaCl), respectively. It indicated that the dry and loose granular embryogenic callus could be selected as somatic salt-tolerant mutant screening material. The results of SSR molecular markers showed that there were 8 pairs of primers in 479 primer combinations in the hippocampus field seedlings, tissue culture regenerated plants and regenerated plant mixed strains screened by different NaCl concentrations (1.2%, 1.6%, 2.0%) Two pairs of primers WXE-30 and WXE-246 were suitable for molecular identification of salt tolerant mutants.