高渗处理对不同基因型大豆未成熟子叶基因枪法转化效果的影响

Effect of Osmotic Treatment on Immature Cotyledons Transformed by Particle Bombardment in Different Genotype of Soybean

为了进一步优化以未成熟子叶为外植体的大豆遗传转化技术体系,研究了高渗剂种类、高渗处理时间,以及轰击后高渗培养时间对不同基因型大豆未成熟子叶基因枪转化效果的影响。结果表明,高渗剂种类极显著地影响体细胞胚数,基因型间体细胞胚胎发生率存在显著差异,而大豆基因型与高渗剂种类间在体细胞胚胎发生率和胚数上都存在极显著的互作。对大豆品种合丰25未成熟子叶进行高渗处理24h时,体细胞胚胎发生率极显著地高于7h和0h处理;胚数在3种高渗时间处理间没有显著差异。基因枪轰击后高渗培养5d和7d时大豆5个基因型间在体细胞胚胎发生率方面存在极显著差异,均以吉育71最高,其次是合丰25和黑农51,绥农28和吉育75较低。吉育71的体细胞胚胎发生率在轰击后高渗培养5d处理中最高,而其它基因型的体细胞胚胎发生率不受轰击后高渗时间的显著影响。

With immature cotyledons as explants, effects of sort of osmotic agent, time of treatment with osmotic agent, osmotic culturing time after transformation by particle bombardment were investigated with the aim to further optimize the transformation system in different genotype of soybean. The results indicated that the number of somatic embryos was affected by the sort of osmotic agent. Also the frequencies of somatic embryogenesis were significantly different among genotypes of soybean. Furthermore, both frequencies of somatic embryogenesis and the number of somatic embryos were affected extremely by the interaction between genotype and sort of osmotic agent. Frequencies of somatic embryogenesis treated with osmotic treatment for 24h were higher than those treated for 7h and Oh for Hefeng 25. No significant difference was observed for the numbers of somatic embryos among 3 kinds of osmotic treatment time. With osmotic treatment for 5d or 7d, frequencies of somatic embryogenesis among 5 genotypes appeared to be significantly different, and the highest one was observed in Jiyu 71 ,followed by Hefeng 25 and He- nong 51. Suinong 28 and Jiyu75 were lower. With osmotic treatment for 5d after transformation by particle bombard- ment,frequencies of somatic embryogenesis for Jiyu 71 was the highest and other genotypes were no significantly affected by osmotic treatment time after transformation by particle bombardment.