2024年2月25日 星期日
黑莓外植体褐化影响因素分析及适宜培养条件筛选
Analysis of influence factors on browning of blackberry explants and selection of suitable culture condition
2009年 第18卷 第3期 页码[63-68]    下载全文[0.9MB]  
摘要

以黑莓(Rubus spp.)品种‘Kiowa’为实验对象,研究了基本培养基类型、外植体类型、枝条暗处理时间、防褐化剂以及光照度对组织培养中外植体褐化及腋芽诱导的影响。结果表明,在供试的6种基本培养基(MS、1/2 MS、B5、1/2 B5、N6和1/2 N6)中,在MS培养基上外植体的褐化率较低(37.78%)、腋芽诱导率最高(67.89%)且生长状况良好;褐化程度与外植体的木质化程度及取材部位均有一定的关系,以1年生半木质化枝条的上部茎段为外植体,褐化率最低(19.67%)、腋芽诱导率较高(91.89%)且生长状况良好;接种前进行一定时间的暗处理对外植体褐化有一定的抑制作用,对枝条暗处理3 d,防褐化效果最佳;在培养基中添加不同类型和浓度的防褐化剂对外植体褐化和腋芽诱导也有一定的影响,以添加3 000 mg.L-1活性炭或200 mg.L-1VC的防褐化效果较好;培养过程中的光照度与外植体褐化也有一定的关系,在1 500 lx光照度下外植体的褐化率较低(15.66%)且腋芽诱导率和生长状况均较好。根据实验结果,筛选出防止黑莓外植体褐化的适宜培养条件为:以1年生半木质化枝条的上部茎段为外植体,15℃暗处理3 d后,消毒并接种在添加了3 000 mg.L-1活性炭或200 mg.L-1VC的MS培养基(含20g.L-1蔗糖、5.6 g.L-1琼脂及0.2 mg.L-16-BA,pH5.8)上,置于光照度1 500 lx的条件下培养。

Abstract

Effects of five factors including types of basic medium and explant, branch dark treatment time, anti-browning agent and illumination intensity on explant browning and axillary bud inducing were investigated using blackberry (Rubus spp.)cultivar `Kiowa'  as experimental object. The results show that on six basic media (MS, 1/2MS,B 5, 1/2B 5,N 6 and 1/2N 6),the explants inoculated on MS medium show a lower browning rate (37.78%), the highest inducing rate of axillary bud(67.89%) and good growth status. The browning degree of explant has acertain relationship with lignification degree and sampling part. The lowest browning rate(19.67%), higher inducing rate of axillary bud(91.89%)and good growth status are obtained using the upper stem segment of semi-lignification branch of one year old as explants. Dark treating for acertain time before inoculating can inhibit the browning of explants and the anti-browning effect of dark treating for 3d is the best. Different types and concentrations of anti-browning agents added in media have some effects on explant browning and axillary bud inducing, and the  anti-browning effects of adding 3000mg·L- 1 activecarbon or 200mg·L- 1 VC are better.Certainly, the browning degree of explant associates with the illumination intensity in culture process, and under 1 500 lx illumination condition, the browning rate is lower(15.66%) and the inducing rate and growth status of axillary bud are better. According to the results, the culture conditions suitable for anti-browning of blackberry explants are as follows: using the upper stem segment of semi-lignification branch of one year old as explants, treating them in dark condition for 3d with 15℃ , then inoculating on MS medium added 3 000mg·L- 1 active carbonor 200mg·L- 1VC(containing 20g·L- 1sucrose, 5.6g·L- 1agarand 0.2mg·L- 1 6-BA,pH 5.8),under 1 500lx illumination condition for culture.

关键词黑莓; 组织培养; 外植体; 防褐化; 腋芽诱导;
Key wordsblackberry(Rubus spp.); tissue culture; explant; anti-browning; axillary bud inducing
作者王小敏,吴文龙,李海燕,于 盱,李维林
所在单位江苏省•中国科学院植物研究所(南京中山植物园),江苏南京210014
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基金项目国家科学技术部农业科技成果转化资金项目(2007GB2C100118); 江苏省农业高技术项目(BG2007311); 江苏省科技基础设施建设计划(BM2008101);