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为了提高小麦远缘杂交单倍体胚植株再生频率,剥取胚龄16～20d的小麦单倍体胚无菌条件下接种到不同配方的培养基。结果表明,直径<500μm的小胚在1号培养基、直径>500μm态的较大胚于2号培养基能一步成苗直接发育成具有丛生芽和健壮根的幼苗;若把丛生幼芽分开后分别转移到1/2 MS基础培养基,则可分化增殖为多株麦苗。小胚培养基配方为:1/2MS+0.1mg.L-12,4-D+0.2mg.L-1BA+300mg.L-1LH(水解乳蛋白)+50g.L-1蔗糖+7g L-1琼脂,pH=5.8,成苗率71%;大胚培养基配方为:1/2 MS+0.1mg.L-12,4-D+0.2mg.L-1BA+30g.L-1蔗糖+7g.L-1琼脂,pH值5.8,成苗率83%以上,以上培养基维生素部分为正常MS培养基维生素加入量的2倍。适宜于小麦单倍体胚再生和繁殖的生长素效果:2,4-D>IBA>IAA,培养基加入2,4-D后经继代培养途径形成单倍体植株生长旺盛,根系发达,增加了单胚再生植株数和植株丛生苗数量,有利于大幅度提高胚成苗率以及染色体加倍效率,从而提高远缘杂交单倍体育种水平,缩短小麦新品种生产周期。
In order to improve the regeneration frequency of distant hybridization haploid embryos in wheat, the haploid embryos of 16 ~ 20d embryo age were stripped and inoculated into the medium of different formulations under aseptic conditions. The results showed that small embryos with diameter less than 500μm could grow directly into No. 1 seedlings and larger embryos with diameter> 500μm in No. 2 medium. After they were transferred to 1/2 MS basal medium, the proliferation can be divided into multiple strains of wheat seedlings. Small embryo culture medium formulation: 1 / 2MS + 0.1mg.L-12,4-D + 0.2mg.L-1BA + 300mg.L-1LH (hydrolyzed milk protein) + 50g.L-1 sucrose + 7g L- 1 agar, pH = 5.8, seedling rate of 71%; big embryo medium formula: 1/2 MS +0.1 mg.L-12,4-D +0.2 mg.L-1BA +30 g.L-1 sucrose + 7g.L-1 agar, pH 5.8, the seedling rate of 83% or more, the above part of the vitamin medium for normal MS medium 2 times the amount of vitamin added. Which is suitable for the auxin regeneration and multiplication of wheat haploid embryo: 2,4-D> IBA> IAA. After adding 2,4-D into the culture medium, the haploid plants grew vigorously through the subculture, the root system developed, Increasing the number of single embryo regenerated plants and the number of plantlet seedlings is conducive to significantly improve the embryo seedling rate and chromosome doubling efficiency, thereby increasing the distant hybridization haploid breeding level and shorten the wheat new variety production cycle.