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高频稳定的组织培养再生体系是开展白三叶遗传转化的重要前提。为进一步优化白三叶再生体系,本研究以白三叶子叶节为外植体,对影响丛生芽诱导、增殖的激素种类(2,4-D、KT、6-BA和NAA)和浓度进行了优化;同时研究了不同基质配比对组培苗移栽成活率的影响。结果表明:在含6-BA 0.5 mg.L-1和NAA0.1 mg·L-1诱导培养基上丛生芽诱导率最高,达68.33%,且生长状态最好;在含IBA 0.2 mg·L-1的1/2MS培养基上诱导生根率为90%以上,平均每株生根数达6条以上;移栽至腐殖土和珍珠岩(2:1)混合基质中的再生苗成活率可达85%。本研究结果可为建立豆科植物转化体系、加快白三叶基因工程改良及创制白三叶新种质奠定基础。
Tissue culture and regeneration system with high frequency stability is an important prerequisite for the genetic transformation of white clover. In order to further optimize the regeneration system of white clover, we studied the effects of 2,4-D, KT, 6-BA and NAA on the production of hormones (2,4-D, KT, 6-BA and NAA) The effects of different matrix ratios on the survival rate of transplanting tissue explants were also studied. The results showed that the highest induction rate was 68.33% on the medium containing 6-BA 0.5 mg.L-1 and NAA 0.1 mg · L-1, The rooting rate was more than 90% on 1 / 2MS medium of -1, and the average number of rooting per plant was 6 or more. The survival rate of regenerated seedlings in mixed medium of humus soil and perlite (2: 1) Up to 85%. The results of this study may lay the foundation for the establishment of transformation system of legumes, accelerating genetic improvement of white clover and establishing new germplasm of white clover.