二次回归正交设计在灰楸离体培养中的应用

为建立灰楸离体繁殖体系,应用二次回归正交设计与SPSS软件分析,建立灰楸组培继代培养增殖系数、增殖芽长度、增殖芽数、愈伤组织横向膨大、愈伤组织纵向膨大、叶数,生根培养生根率、根长、发根数、发芽数、芽长等回归方程,通过对各方程进行模拟寻求,确定6-BA、IBA、NAA在继代与生根培养中的用量范围。结果表明:灰楸组培继代培养6-BA用量的最佳范围为1.343~1.356 mg·L~(-1),IBA用量在0.11~0.294 mg·L~(-1);一定质量浓度的6-BA在继代培养中起主要的诱导分化作用;IBA对诱导分化芽数有显著的回归关系,与其它指标无显著关系;分化芽叶数的多少与6-BA与IBA无显著关系。灰楸组培生根培养IBA用量的最佳范围为0.272~0.329 mg·L~(-1),NAA为0.027~0.033 mg·L~(-1);一定质量浓度的IBA主要促使根系与芽长生长,对生根率与发芽数无显著作用,NAA起着主要的诱导根系发生作用,与生根率、发芽数有显著的回归关系;生根苗叶数的多少与6-BA与IBA无显著关系;IBA用量在0.272~0.329 mg·L~(-1)、NAA在0.027~0.033 mg·L~(-1)范围内,灰楸组培生根率可达96%以上,可靠性为95%。灰楸离体培养适宜的继代培养基为DKW+6-BA 1.343~1.356 mg·L~(-1)+IBA 0.11~0.294 mg·L~(-1)+琼脂4.5 g·L~(-1)+蔗糖25 g·L~(-1)(p H=5.8);生根培养基为1/2MS+IBA 0.272~0.329 mg·L~(-1)+NAA 0.027~0.033mg·L~(-1)+琼脂5.0 g·L~(-1)+蔗糖10 g·L~(-1)(p H=5.8)。

Application of Quadratic Regressive Factorial Experiment to in vitro Culture of Catalpa fargesii

We used quadratic regressive factorial experiment and SPSS software to analyze growth coefficient,bud length,proliferation bud number,callus enlargement and leaf number of Catalpa fargesii during subculture,and optimize concentrations of IBA and 6-BA in subculture medium.We obtained a serial regression equations,and found the best dosage of IBA and 6-BA to subculture the callus of C.fargesii through simulation and search of the equations.A certain concentration of 6-BA could promote the bud differentiation in subculture of C.fargesii,and the best dosage range of 6-BA was 1.343-1.356mg·L-1.IBA had a significant regression relationship with bud differentiation,and the best dosage range of IBA was 0.11-0.294mg·L-1,but there was no significant relationship with other indicators.The number of bud differentiation had no significant relationship with 6-BA and IBA.We also built regression equations through the analyzing the rooting rate,root length,root number,sprouting number and bud length in rooting medium of C.fargesii.We obtained the best dosage of IBA and NAA using simulation and search of these equations.The best dosage range of IBA was 0.272-0.329mg·L-1,and the best dosage range of NAA was 0.027-0.033mg·L-1 in rooting medium of C.fargesii.A certain concentration of IBA mainly promoted the growth of root and bud length,but there was no significant effect with root number and sprouting number.NAA mainly induced the root,and had a significant regression relationship with the rooting rate and sprouting number.However,the leaf number of rooted seedlings had no significant relationship with 6-BA and IBA.When the dosage range of IBA was 0.272-0.329mg·L-1,and NAA was 0.027-0.033mg·L-1,the rooting rate of C.fargesii tissue culture reached more than 96％and the reliability was 95％.