榆叶梅高效成熟胚培养成苗体系的建立

Establishment of a Culture System for Mature Embryos of Prunus triloba Lindl.

以榆叶梅(Prunus triloba Lindl.)成熟胚为外植体,构建榆叶梅稳定高效的胚培养成苗体系,测定萌发率、平均株高、生根率、平均数、平均根长、愈伤率和叶片形态等指标,设置MS、1/2 MS、WPM等3个基础培养基,筛选榆叶梅胚培养成苗的最适培养基;利用6-BA、NAA、IBA等3种植物生长调节剂和活性炭进行胚的萌发和生根试验,其中IBA、NAA均设0.05、0.1、0.2 mg/L等3个水平,6-BA设1、2 mg/L两个水平,统计胚萌发和生根情况,筛选榆叶梅胚培养成苗体系最佳培养基类型;在最适合榆叶梅胚培养生长培养基基础上,再设置不同低温处理时间(0、20、40、60、80 d)进行愈伤组织诱导及分化实验,统计愈伤率和叶片形态情况;最后分析不同子叶保留程度(完整子叶、1/2子叶和1/4子叶)对胚培养萌发和生根的影响。结果表明,MS培养基为榆叶梅胚的萌发和上胚轴生长的最适培养基;综合各项试验的最佳影响因子,榆叶梅在MS+6-BA 1.0 mg/L+IBA 0.1 mg/L+NAA 0.1 mg/L+活性炭500 mg/L培养基上萌发率和生根率最高,分别为83.33%、86.67%;4℃冷藏60~80 d为最适宜榆叶梅胚培养发育和正常生长的时间,冷藏时间80 d时萌发率达100%,生根率为93.33%,叶片绿色平展;保留完整子叶有利于促进上胚轴的伸长,保留1/4子叶有利于促进下胚轴的伸长。本研究筛选出最适宜榆叶梅胚的培养成苗培养基为MS+6-BA 1.0 mg/L+IBA 0.1 mg/L+NAA 0.1 mg/L+活性炭500 mg/L。

Using mature embryos of Prunus triloba Lindl.as explants,a stable and efficient embryo culture seedling system was established.The germination rate,average plant height,rooting rate,average root length,callus rate and leaf morphology were determined,and MS,1/2 MS and WPM were set.The optimum medium for the seedling cultured from Prunus triloba Lindl.embryo was selected.The germination and rooting tests of embryos were carried out using plant growth regulators 6-BA,NAA,IBA and activated carbon.IBA and NAA levels were 0.05,0.10,0.20 mg/L and 6-BA setting 1 and 2 mg/L 2 levels,the germination and rooting of embryos were counted,and the best medium type of the seedling system was screened.On the basis of the most suitable growth medium for Prunus triloba Lindl.embrasure culture,callus induction and differentiation experiments were conducted at different low temperature treatment times(0,20,40,60,80 days),and callus rate and leaf morphology were collected.Finally,the effects of different degrees of cotyledon retention(complete cotyledon,1/2 cotyledon and 1/4 cotyledon)on germination and rooting of embryo culture were analyzed.The results showed that MS medium was the most suitable medium for the germination and epicotyl growth of Prunus triloba Lindl.embryo.The germination rate and rooting rate were the highest on MS+6-BA 1.0 mg/L+IBA 0.10 mg/L+NAA 0.10 mg/L+activated carbon 500 mg/L,which were 83.33%and 86.67%,respectively.The most suitable time for Prunus triloba Lindl.embryo development and normal growth was refrigerated at 4℃from 60 to 80 days,and when the refrigeration time was 80 days,the germination rate reached 100%,the rooting rate was 93.33%,and the leaves were green and flat.Keeping intact cotyledon was more conducive to promoting the elongation of epicotyl,and keeping 1/4 cotyledon was more conducive to promoting the elongation of hypocotyl.In this study,the most suitable seedling medium was MS+6-BA 1.0 mg/L+IBA 0.10 mg/L+NAA 0.10 mg/L+activated carbon 500 mg/L.