小麦成熟胚组织培养再生体系的优化及高再生率基因型的筛选

Optimization of Regeneration System of Tissue Culture from Mature Embryos and Screening of Wheat Genotypes with High Regeneration Frequency

为了优化小麦成熟胚组织培养再生体系,并从优良小麦品种中筛选出高再生率基因型,选用20个小麦品种为材料,研究了4种胚的不同处理方式对成熟胚脱分化率及形成愈伤组织质量的影响,6种不同激素配比对愈伤组织分化再生的影响及2种生根培养基的生根效果;并对参试品种中的高再生率基因型进行了筛选。结果表明,用纵半切的全胚进行愈伤组织诱导形成的愈伤组织质量最好;小麦成熟胚愈伤组织在以下3种激素配比,即:B.IAA 1.0 mg.L-1+ZT 2 mg.L-1;E.2,4-D 0.1 mg.L-1+ZT 10 mg.L-1;F.6-BA 1 mg.L-1+NAA 0.3 mg.L-1+KT 4 mg.L-1的培养基上分化再生成苗率较高;再生苗在不加生长素的1/2MS培养基上生根效果很好。在供试的20个小麦品种中,有12个小麦品种的再生成苗率达到20%以上,其中西农928、宁春16和西农889的再生成苗率分别高达77.6%、52.7%和36.9%。B分化培养基对大部分基因型愈伤组织的分化培养具有比较好的通用性。本研究从黄淮麦区推广的小麦品种中筛选出的一些高再生率的基因型可以用作该地区小麦转基因研究的受体材料。

This study aimed to optimize regeneration system of tissue culture from mature embryos in wheat and to select genotypes with high regeneration frequency from some good wheat varieties.20 wheat varieties were used to investigate the effects of six kinds of different hormone ratio on callus regeneration and the effects of two culture mediums for rooting on root generation.Finally,the genotypes with high regeneration frequency were selected from 20 wheat cultivars investigated.The results showed that among the six kinds differentiation mediums with different hormone ratio for callus regeneration,3 kinds of differentiation mediums,i.e.medium B of IAA 1.0 mg·L-1 and ZT 2 mg·L-1;medium E of ZT 10 mg·L-1and 2,4-d 0.1 mg·L-1 and medium F of 6-BA 1 mg·L-1,NAA 0.3 mg·L-1 and KT4 mg·L-1,were suitable for mature embryo regeneration of all the three wheat genotypes,and the regeneration rate was much higher than those of the other three treatemnts.Rooting culture medium of 1/2 MS without auxin was more likely to prompt buds to generate roots.Twelve of the 20 wheat cultivars investigated obtained regeneration rate more than 20%,among which the regeneration rate of Xinong 928,Ningchun 16 and Xinong 889 were as high as 52.7%,77.6% and 36.9 %,respectively.Differentiation medium B is adaptable for callus culture of most genotypes.A regeneration system of tissue culture from wheat mature embryos adaptable for most genotypes was well established and genotypes with high regeneration frequency were selected from wheat cultivars widely planted in Shaanxi province and HuangHuai wheat area in the present study.These wheat cultivars can be used as the receptor varieties in wheat transgenic research.