'早红'草莓高效遗传转化受体系统的建立

本文以草莓主栽品种'早红'组培苗离体叶片和叶柄为外植体,进行叶龄、暗培养、植物生长调节剂配比及抗生素敏感性研究,建立草莓高效遗传转化的受体系统.在含3.0 mg/L 6-BA与0.1 mg/L 2,4-D的MS培养基上,30 d叶龄的叶片再生频率高达98.31%,平均每叶片再生芽数5.09个,叶柄切段的再生频率为89.25%,平均每叶柄切段再生芽数4.92个,叶片的再生频率略高于叶柄;不定芽在含0.2 mg/L 6-BA与0.2 mg/L GA_3的MS继代培养基上培养成苗.将生长状态良好的不定芽转至含0.2 mg/L IBA的1/2 MS培养基上生根,生根率达100%,平均生根数量16.27条,平均根长1.85 cm.抗生素敏感性试验表明,草莓外植体适宜的卡那霉素选择压力为25 mg/L,头孢霉素的筛选浓度为300mg/L.本研究建立的再生体系可作为草莓遗传转化的受体系统.

Establishment of Recipient System of High Frequency for Genetic Transformation of 'Zaohong' Strawberry

An efficient regeneration system was established from the explants of leaves and petioles of‘Zaohong'strawberry(Fragaria ananassa Duch.).The effects of different seedling age,dark incubation,growth regulators with different ratio and the sensitivity of explants to the antibiotics on bud differentiation and plant regeneration were comparatively investigated in this study.After being cultured for 30 days on the MS medium supplemented with 3.0 mg/L 6-BA and 0.1 mg/L 2,4-D.The frequency of bud differentiation reached to 98.31%,and the number of regenerated shoots per leaf disc was 5.09.The differentiation frequency of adventitious buds of petiole segments was 89.25%.the number of shoots per petiole segment was 4.92.It is obviously that the differentiation potential of leaf discs Was much higher than that of the petiole segments.The adventitious buds subsequently grew up to plantlets on the MS medium supplemented with 0.2 mg/L 6-BA and 0.2 mg/L GA_3.After well-growing plantlets were transplanted on the 1/2 MS medium added with 0.2 mg/L IBA,100%of rooting frequency was obtained.The average number of roots per plantlet was 16.27,and the average length of roots reached 1.85 cm.The examination of sensitivity to antibiotics suggested that the appropriate screening concentrations of Kanamycin and Cefotaxime sodium for transformation of strawberry were 25 mg/L and 300 mg/L,respectively.This established plant regeneration system might be used as a recipient system for genetic transformation of strawberry.