白术愈伤组织诱导及丛生芽增殖培养基优化研究OA

To establish an efficient in vitro regeneration and rapid propagation system for Atractylodes macrocephala Koidz.and solve the problems of low efficiency and cultivar degradation in traditional propagation methods,stems and leaves of aseptic A.macrocephala seedlings were used as explants in this study.Single-factor experiments were conducted to screen suitable conditions for callus induction,and an L9(34)orthogonal experimental design was adopted to systematically optimize the effects of four key factors(6-BA,NAA,TDZ and sucrose)on the differentiation and proliferation of cluster buds,with supporting rooting and transplanting procedures established.The results showed that stem explants performed better than leaf explants in callus induction.The optimal callus induction medium was MS medium supplemented with 2.0 mg·L-1 2,4-D,0.1 mg·L-1 6-BA and 35g·L-1 sucrose,under which the callus induction rate reached 92.50％and the embryonic callus rate reached 81.67％.Range analysis and variance analysis of the orthogonal test indicated that the order of influence of the four factors on the proliferation coefficient of cluster buds was 6-BA>NAA>TDZ>sucrose,among which 6-BA and NAA had significant effects(P<0.05).The optimal differentiation and proliferation medium was MS medium containing 2.0 mg·L-1 6-BA,0.2 mg·L-1 NAA,0.2 mg·L-1 TDZ,35g·L-1 sucrose and 7g·L-1 agar.On this medium,the cluster bud differentiation rate reached 95.56％,the proliferation coefficient reached 6.42,and the average bud length was 2.17 cm,with stout buds,dark green leaves and no vitrification.The optimal rooting medium was 1/2 MS medium supplemented with 0.5 mg·L-1 IBA and 30g·L-1 sucrose,with a rooting rate of 93.33％and an average root length of 3.42 cm.After acclimatization,plantlets were transplanted into a substrate of peat soil and perlite at a volume ratio of 2:1,with a survival rate of 89.17％.This study established a complete in vitro regeneration system for A.macrocephala from callus induction to transplant survival,which can provide technical support for large-scale propagation of elite seedlings and germplasm innovation of A.macrocephala.