黑穗醋栗(Ribes nigrum L.)未受精胚珠培养诱导体细胞胚状体和植株再生的研究

STUDIES ON INDUCTION OF SOMATIC EMBRYOS AND PLANTLET REGENERATION FROM UNFERTILIZED OVULES IN VITRO CULTURE OF BLACK CURRANT (RIBES NIGRUM L.)

本文报告了黑穗醋栗(Ribes nigrum L.)三个栽培品种中的薄皮黑豆未受精胚珠(花粉发育到单核晚期)在MS基本培养基附加植物激素BA,2.4-D和GA_3中形成了体细胞胚状体。长度约为0.5—1.0cm大小的胚状体在生根培养基中可以形成完整的再生植株。经过筛选得到了体细胞胚性愈伤组织无性系,继代培养二年多仍能保持胚状体形成能力。试验结果表明:诱导体细胞胚胎发生受品种和接种时期的影响,适宜的接种时期为花粉发育到单核晚期。培养基中的BA为诱导胚胎发生和保持胚性愈伤组织无性系所必需。

This paper first reported that Baopiheidou, one of three cultivarsin this experiment was able to produce adventive embryos when theunfertilized ovules of black currant (Ribes nigrum L.) were inoculated onMS medium supplemented with plant hormones BA, 2.4-D,GA_3.Thesomatic embyogenic calli were inductedo in basal medium MS supplementedwith 5.0-3.0mg l^(-1) BA, 2.0mg l^(-1) 2.4-D or 1.0mg l^(-1) GA_3. After cultu-ring the unfertilized ovules about 25 days on the induction medium, the calli were trasfered to the embryo formation medium for the embryodevelopment. The adventive embryos were observed after culturing about20 days on the embryo formation medium (Basal medium MS + 3.0mg l^(-1)BA + 1.0mg l^(-1) GA_3 + 3% sucrose. ). Somatic embryo with 0.5-1.0 cmlength was able to form whole plantlet in medium MS + 2.0mg l^(-1) IBA +2.0mg l^(-1) GA_3 + 1% sucrose. The experiments showed: The variety andinduction stage of inoculated ovules affected the somatic embryogenesis.The somatic embryogenesis was inducted when ovules at the uninuleatestage were inoculated. Its induction frequency was 3.3%-5.0%. BA in themedium was necessary for the induction of somatic embryogenesis andembryo development. We also had established somatic embryogenic callusclone directly from somatic cell of black currant cultivar and maintainnedmore than two years with the avaibility of embryogenesis, It can be usedas a favourable material for the micropropagation, mutation selection ofcell suspension culture, protoplast isolation and culture and genetic man-ipulation in black currant.