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

作　　者：王小敏[1] 吴文龙[1] 李海燕[1] 于盱[1] 李维林[1] 

机构地区：[1]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

出　　处：《植物资源与环境学报》2009年第3期63-68,共6页Journal of Plant Resources and Environment

基　　金：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)

摘　　要：以黑莓(Rubusspp.)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。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.） cuhivar ＇ Kiowa＇ as experimental object. The results show that on six basic media（MS, 1/2 MS, B5, 1/2 B5, N6 and 1/2 N6）, 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 a certain 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 a certain time before inoculating can inhibit the browning of explants and the anti-browning effect of dark treating for 3 d 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 3 000 mg · L^ -1 active carbon or 200 mg· 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 antibrowning 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 3 d with 15 ℃. then inoculating on MS medium added 3 000 mg · L^- 1 active carbon or 200 mg - L^- 1 Vc （ containing 20 g · L^-1 J sucrose, 5.6 g · L^- 1 agar and 0.2 mg · L^-1 6-BA, pH 5.8 ）, under 1 500 lx illumination condition for culture.

关 键 词：黑莓 组织培养 外植体 防褐化 腋芽诱导 

分 类 号：Q943.1[生物学—植物学] S663.2[农业科学—果树学]