大豆叶柄离体再生体系研究

Research on plantlet regeneration from petiole explants in soybean

[目的]建立大豆叶柄外植体的离体再生体系,为大豆遗传转化体系的改进和提高提供技术基础。[方法]以大豆品种‘K06-82’无菌苗叶柄为外植体,比较不同种类的植物生长调节剂及其组合对叶柄愈伤组织诱导、不定芽分化、不定芽伸长及生根等过程的影响,建立大豆叶柄离体再生体系。[结果]大豆品种‘K06-82’的叶柄愈伤组织形成的适宜培养基为改良MS+2.0 mg·L^(-1)α-萘乙酸(NAA),愈伤组织形成率为74.21%,愈伤组织密度为0.72 g·cm^(-3);不定芽分化的适宜培养基为改良MS+2.0 mg·L^(-1)6-苄氨基嘌呤(6-BA)+0.2 mg·L^(-1)NAA,分化率为49.82%,平均不定芽数2.98个;不定芽伸长的适宜培养基为改良MS+1.0 mg·L^(-1)赤霉素(GA3),伸长比率为35.62%,平均芽长为2.01 cm;不定根诱导的最适培养基为1/2改良MS+0.8 mg·L^(-1)吲哚丁酸(IBA),生根率和生根苗的移栽存活率分别达到88.32%和97.80%。[结论]初步建立了大豆叶柄外植体的离体再生体系。

[ Objectives ] The purpose of this research is to establish the plantlet regeneration system in soybean. [ Methods ] By using the aseptic seedling petioles of soybean variety ＇ K06-82 ＇ as explants, the effects of different plant growth regulators and their combinations on callus induction and adventitious bud differentiation, adventitious bud elongation and rooting process were compared to establish the regeneration system for soybean stalk in vitro. [ Results ] The results showed that the suitable medium for callus formation from soybean＇ K06-82＇ petiole explants was modified MS＋2.0 mg· L-1 a-naphthylacetie acid （NAA）, on which the callus formation rate was 74.21%, and the density of callus was 0.72 g·cm-3. The optimal medium for the adventitious buds differentiation was modified MS＋2.0 mg.L-1 6-benzylamino purine（6-BA）＋0.2 mg-L-l NAA,giving a differentiation frequency of 49.82% and an average number of adventitious buds 2.98. The suitable medium for adventitious bud elongation was modified MS＋ 1.0 mg·L-L gihberellin（ GA3 ）. The elongation efficiency was about 35.62%, and the average shoot length was 1.01 era. Half strength modified MS medium supplemented with 0.8 mg·L-1 indole butyric acid （IBA）was the suitable medium for inducing adventitious roots, producing a rooting percentage of 88.32% and a seedling transplanting smwival rate of 97.80%. [ Conclusions ] In this study, the in vitro culture and plantlet regeneration system for soybean petiole explants are established, which will provide the theoretical and technical basis for the establishment of a more perfect soybean genetic transformation system.