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水稻不同品种组培再生和转基因频率研究 被引量:14

Investigation of Regeneration and Transformation Frequencies Generated from Different Genotypes in Rice( Oryza sativa L. )
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摘要 本研究选取21个(包括16个籼稻品种和5个粳稻品种)在农业生产上具有重要应用价值的水稻品种,对它们进行成熟胚的组培再生能力以及转基因效率的考察。结果表明:以NBm诱导培养基进行愈伤诱导时,各水稻品种的愈伤诱导率介于30.37%和72.46%之间;以NBm和DL3分化培养基对各水稻品种不同日龄愈伤分别进行植株再生培养,并以植株再生率(plant regeneration rate,RR)和每粒愈伤平均出苗数(the mean number of regenerated shoots,NRS)对再生能力进行考察时,不同水稻品种的组培再生能力表现较为复杂,同时受到基因型、愈伤日龄和培养基成分的重大影响。在NBm分化培养基上,以日龄15d的T0974愈伤RR(84.00%)和NRS(2.96)最高,在DL3分化培养基上,以日龄15d的T0974愈伤RR(73.33%)和粤泰B愈伤NRS(3.55)分别为最高。此后,利用根癌农杆菌介导的转基因方法对其中12个水稻品种进行成基因转化实验,经PCR验证后发现,共成功实现7个品种的&基因转化,各水稻品种的转化效率介于0~25%之间,且与其再生能力相关。 In this study, a total of 21 elite rice varieties, 16 indica and 5 japonica, were selected and investigated for the regeneration and transformation frequencies generated from tissue culture experiments. The results showed that i) the callus induction rate in the 21 cultivars were between 30.37% and 72.46% when cultured on the NBm callus induction medium; ii) regeneration ability of 21 varieties was associated with genotypes, the age of calli and medium composition. To974 callus of age 15 days had the highest number of plant regeneration rate (RR) (84.0%) and the mean number of regenerated shoots (NRS) (2.96) among all tested cuhivars on NBm differentiation medium ; and both To974 callus of age 15 days and Yuetai B callus of age 15 days showed the highest number of RR (73.33%) and NRS (3.55), respectively, on DL3 differentiation medium. Agrobacterium-mediated transformation of Bt gene was then performed on 12 varieties and transgenic plants were obtained from 7 varieties after PCR identification of the target gene. Different frequencies of transformation in different rice varieties were ranged from 0 to 25% and corresponded with different regeneration frequencies of the varieties.
出处 《核农学报》 CAS CSCD 北大核心 2013年第12期1817-1827,共11页 Journal of Nuclear Agricultural Sciences
基金 转基因生物新品种培育重大专项(2009ZX08001-008B)
关键词 水稻 愈伤 再生能力 转化效率 Rice(Oryza sativa L. ) Callus Regeneration ability Transformation efficiency
作者简介 李素娟,女,主要从事植物生物技术和作物遗传改良研究。E-mail:sujuanli2001@163.com 樊秀霞,女,主要从事植物抗逆遗传改良研究。E-mail:754674253@qq.com。同为第一作者。 通讯作者:钱前,男,研究员,主要从事水稻经典遗传、分子遗传育种研究。E-mail:qianqianl88@hotmail.com
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