Artificial Propagation Of Anoectochilus Roxburghii (Wall.)Lindl., A Rare Medicinal Plant In China

Anoectochilus roxburghii (Wall.) Lindl.(family Orchidaceae) is a well known medicinal and ornamental plant. Its wild resources is confronted with exhaustion. Artificial propagation is the only way for industrial application of this plant. In this paper, the techniques of tissue culture were applied to improve its propagation, The results are as follows:1. Protocorm proliferation of A. roxburghiiL9(33) orthogonal experiments were conducted to optimize protocorm culture medium with, factors and levels as follows:medium(MS,1/2MS, B5), NAA (0.3,0.6and0.9mg·L-1) and BA (0.3,0.6and0.9mg·L-1). Protocorm was developed in1/2MS added with1.0g/L AC,30g/L sucrose and6.0g/L agar, and cultured under24-28℃,1500～2000Lux(12h/d) for25d. The obtained protocorms were inoculated into the9media and raised for42d, The medium (0.6mg·L-1NAA+0.6mg·L-1BA+1/2MS) was selected as protocorm proliferation medium according to the reproductive rate.2. Protocorm differentiation of A. roxburghiiL9(33) orthogonal experiments with, factors and levels of medium (MS,1/2MS, B5), NAA (1,2and3mg·L-1) and BA (0.1,0.2and0.4mg·L-1) was carried out to optimize protocorm differentiation medium. Protocorm was inoculated and cultured under24-28℃癈,1500-2000Lux (12h/d) for28d. Leaves developed on some media, and1/2MS added with2.0mg/L NAA and0.2mg/L BA was used as protocorm differentiation medium according to the differentiation rate.3. Protocorm rooting of A. roxburghiiAccording to the rooting rate, protocorm rooting medium was selected as1/2MS supplemented with0.6mg/L NAA and10%banana juice by orthogonal experiments with factors and levels as follows:medium (MS,1/2MS, B5), NAA(0.3,0.6and0.9m·L-1), nutrient additives (10%banana juice,10%potato juice,10%coconut juice). 4. Transplantion of A. roxburghiiThe transplanting condition of A. roxburghii was optimized by L9(33) orthogonal experiments. The factors and levels were as follows:medium (perlite:silica:garden soil=1:1:4, perlite:silica:garden soil=1:1:6, perlite:silica:garden soil=1:1:8), nutrient solution (500,1000and1500times) and acclimation time(10d,20d,30d), According to the survival rate of plantlet, the optimized transplanting protocol was as follows: the rooted plantlets were transplanted into medium (perlite:silica:garden soil=1:1:6), which added with1000times nutrient solution, and then acclimated for20d.In conclusion, the propagation of A. roxburghii protocorm was optimized in this thesis, including protocorm proliferation, differentiation, rooting and transplantation. These results lay a good foundation for industrial application of A. roxburghii.