正交试验法在荔枝体胚诱导过程中的应用

Application of Orthogonal Experiment on Induction of Somatic Embryogenesis in Litchi

应用正交试验方法,研究MS培养基中附加的BA、KT、NAA、2,4-D以及琼脂和蔗糖浓度对荔枝胚性愈伤组织体胚诱导的影响。结果表明,BA 0.5 mg.L-1+KT 0.5 mg.L-1组合有利于荔枝胚性愈伤组织诱导体胚,NAA及2,4-D对体胚诱导效果不明显;在高浓度蔗糖(60 g.L-1)和琼脂(10 g.L-1)作用下,荔枝体胚诱导率较高,其中琼脂的作用效果明显高于蔗糖。通过正交试验方法选用MS+BA 0.5 mg.L-1+KT 0.5 mg.L-1+LH 500 mg.L-1+蔗糖60 g.L-1+琼脂10 g.L-1组合,荔枝胚性愈伤组织诱导的体胚达9.05百个.g-1,是MS基本培养基的4.6倍,减少了试验次数,提高了荔枝体胚诱导的工作效率与准确性。

The mediums adapted to somatic embryogenesis induction of litchi embryogenic callus were selected by using orthogonal experiment design. The benzyladenine （ BA）, kinetin （KT）, α - naphthylacetic acid （NAA）, 2, 4 -dichlorophenoxyacetic acid （2, 4- D）, agar and sucrose were used to compare their effects on inducing somatic embryogenesis in litchi. The results showed that the combination of BA 0. 5 mg·L^- 1and KT 0. 5 mg· L^- 1 were propitious to inducing somatic embryogenesis. Both NAA and 2, 4 -D had no effect on somatic embryogenesis induction. However, agar 10 g · L^-1 and sucrose 60 g · L^-1enhanced the induction of somatic embryogenesis, and the effect of agar was higher than sucrose. MS ＋ BA 0. 5 mg· L^-1 ＋ KT 0. 5 mg· L^-1 ＋ LH 500 mg·L^-l ＋ sucrose 60 g · L^-l ＋ agar 10 g · L^-1was selected as the best medium for litchi somatic embryogenesis induction by using orthogonal experiment. The number of induction was 9.05 hundred per gramme, which was 4. 6 times in the basic medium. Working efficiency and accuracy were increased in the induction of litchi somatic embryogenesis.