赣南脐橙无菌苗生产的组培脱菌与微芽嫁接快繁技术

Rapid Propagation of Disease-free Citrus sinensis Osbeck vc. Newhall Production Using Debacterization and Microshoot tip grafting in vitro

以柑橘良种赣南脐橙带芽茎段为外植体,枳壳无菌实生幼苗作为砧木,探讨了消毒灭菌方法、植物生长调节剂组合、热处理温度和时间及茎尖尺寸大小对其无菌体系的建立、丛生芽的诱导与增殖、脱菌及微嫁接的影响。结果表明:采用70%酒精消毒30 s,无菌水冲洗3次,再用0.1%升汞加吐温-80消毒15 min,可获得成功率为83.20%的有生命力无菌培养材料;以MS+6-BA 1.0 mg·L-1+NAA 0.5 mg·L-1为最佳芽诱导培养基,诱导率达89.72%;最佳丛生芽增殖培养基为MS+6-BA 2.0 mg·L-1+NAA 0.4 mg·L-1+GA3 0.1 mg·L-1,增殖率最高达394.26%;采用40℃高温处理40 min的0.6 mm茎尖进行微嫁接,成活率为63.32%,脱菌率为93.21%。

The effects of disinfection and sterilization methods,plant growth regulator combination,heat treatment temperature and time,and the size of stem tip on the establishment of sterile system,induction and proliferation of cluster buds,debacterization and microshoot tip grafting were discussed for(Citrus sinensis(L.)Osbeck)in Gannan region.The results showed that 83.20%of the viable sterile culture materials could be obtained by sterilizing with 70%alcohol for 30 seconds,washing with sterile water for 3 times,and then sterilizing with 0.1%mercuric chloride and Tween-80 for 15 minutes.Taking MS+6-BA 1.0 mg·L-1+NAA 0.5 mg·L-1 as the optimal bud induction medium,the induction rate reached 89.72%.The best multiplication medium for cluster buds was MS+6-BA 2.0 mg·L-1+NAA 0.4 mg·L-1+GA3 0.1 mg·L-1,with the highest multiplication rate of 394.26%.The survival rate was 63.32%and the bacteria and virus-free rate was 93.21%when the stem tip of 0.6 mm was treated at 40℃for 40 minutes.