杂交榛愈伤组织诱导和原生质体分离

Callus Induction and Protoplast Isolation from Hybrid Hazel

利用组织培养技术和原生质体分离技术研究极早熟杂交种榛子,为进一步创造新种质资源并应用于育种实践提供基础性研究材料。结果表明,对1年生茎段、叶片和当年生子叶3种外植体而言,当年生子叶愈伤组织诱导率最高,最适合的植物生长调节剂组合为2,4-D0.5mg·mL-1+6-BA1.0mg·mL-1+KT2.0mg·mL-1;以花药为外植体时,最佳植物生长调节剂组合为2,4-D1.0mg·mL-1+6-BA1.0mg·mL-1;体细胞愈伤组织和花药愈伤组织原生质体分离的最佳酶组合分别是0.01g.mL-1纤维素酶+0.03g.mL-1果胶酶、0.01g.mL-1纤维素酶+0.02g.mL-1果胶酶,最适酶解时间分别为6～7h、5h;与体细胞愈伤组织相比,花药愈伤组织很难分离出大量原生质体,不适合做原生质体的分离材料。

An experiment was conducted to study the tissue culture and protoplast isolation of the early maturing hybrid hazel in order to provide basic research materials for the creation of new germplasm resources and the application to breeding practice.Results showed that the callus induction frequency of current-year cotyledons was the highest compared with one-year-old hazel stem segments and leaves.The optimal plant growth regulators were the combination of 2,4-D 0.5mg·L-1,6-BA 1.0mg·L-1 and KT 2.0mg·L-1 when one-year-old hazel stem segments,leaves and current-year cotyledons were used as explants,while the combination of 2,4-D 1.0mg·L-1 and 6-BA 1.0mg·L-1 was suitable for callus induction when the anthers was used as explants.The optimum enzyme and hydrolysis time were 0.01g·mL^-1 cellulase ＋ 0.03g·mL^-1 pectase and 6-7h for protoplast isolation of somatic cell callus and 0.01g·mL^-1 cellulase＋0.02g·mL^-1 pectase and 5h for that of anther callus.Compared with the somatic cell callus,it was difficult for the anther callus to isolate a large number of protoplasts;therefore it is not suitable as a source material for protoplast isolations.