响应面法优化珍珠相思不定芽诱导及增殖培养条件

Response Surface Methodology for Optimization of Adventitious Bud Induction and Proliferation Cultures of A.podalyriifolia

本试验以珍珠相思带腋芽的茎段为外植体,采用响应面法优化珍珠相思诱导及增殖培养基,以期提高珍珠相思外植体的诱导率以及不定芽的增殖系数,为珍珠相思建立组培快繁体系提供参考依据。通过单因素试验确定不定芽诱导及增殖的基本培养基以及激素添加范围,并利用响应面法中的CCD设计进行优化分析。结果表明,由单因素试验法分析MS培养基可作为珍珠相思不定芽诱导培养的最佳基本培养基;不定芽诱导与增殖培养基激素添加范围分别为6-BA 1.5~2.5 mg/L,NAA 0.1~0.3 mg/L,2,4-D 0.10~0.20 mg/L、6-BA 0.6~1.0 mg/L,NAA 0.3~0.5 mg/L,GA30~0.2 mg/L。再通过响应面法得到最佳诱导培养基、增殖培养基分别是MS+6-BA 2.00 mg/L+NAA 0.2 mg/L+2,4-D 0.14 mg/L、MS+6-BA 1.00 mg/L+NAA 0.40 mg/L+GA30.06 mg/L,其实际诱导率、增殖系数分别为89.36%、3.54,与预测值十分接近,说明回归方程与真实试验拟合度较好。本研究创新性应用响应面方法优化珍珠相思不定芽诱导及增殖培养条件,减少了实验次数,节省了试验成本和时间,能广泛应用于工厂化育苗,为珍珠相思的良种选育及推广种植奠定理论和实践基础。

In this experiment,the axillary buds of Acacia podalyriifolia were used as explants.Response surface methodology was used to optimize the medium for induction and proliferation of Acacia podalyriifolia,with a view to increasing the induction rate of Acacia podalyriifolia explants and the proliferation coefficient of adventitious buds.The tissue culture rapid propagation system provides a reference basis.The effects of basic medium and different hormone concentrations(6-BA,NAA,2,4-D,GA3)on the induction and proliferation of adventitious buds were determined by single-factor experiments,and optimized analysis was performed using the CCD design in response surface methodology.The results showed that the analysis of MS medium by single factor test could be used as the best basic medium for adventitious bud induction culture of Acacia podalyriifolia.0.1~0.3 mg/L,2,4-D0.10~0.20 mg/L,6-BA 0.6~1.0 mg/L,NAA 0.3~0.5 mg/L,GA30~0.2 mg/L.The best induction medium and proliferation medium were obtained by response surface method.MS+6-BA 2.00 mg/L+NAA 0.2 mg/L+2,4-D0.14 mg/L,MS+6-BA 1.00 mg/L+NAA 0.40 mg/L+GA30.06 mg/L,the actual induction rate and proliferation coefficient were 89.36%and 3.54,which are very close to th e predicted values,indicating that the regression equation fits the real test well.In this study,response surface methodology was innovatively applied to optimize the conditions of Acacia podalyriifolia adventitious bud induction and proliferation culture,which effectively reduced the number of experiments,saved the cost and time of experiments,and facilitated industrial seedling cultivation.It lays a theoretical and practical foundation for the breeding and popularization of Acacia podalyriifolia.