| Fructus Amomi, one of south medicine, is the dried ripe fruit of Zingiberaceae Amomum villosum Lour.,Amomum villosum Lour. var. xanthioides T.L.Wu et Senjen or Amomum longiligulare T.L. Wu, used to resolve dampness for treating damp accumulation in the spleen and stomach, to warm the middle for treating vomiting and diarrhea due to deficiency-cold of the spleen and stomach, and to prevent miscarriage in cases of threatened abortion. The best Fructus Amomi is the dried ripe fruit of Zingiberaceae Amom um villosum Lour. producted Yangchun in Gangdong. It suffers serious diseases and varieties degradation causing sharply reduction of Fructus Amomi.Gene engineering had used in the medicinal plant in recent years, such as foreign gene was expressed in Lilium brownie, Pogostemon cablin(Blanco) Benth. and Houttuynia cordata Thunb. The studies achieved remarkable results that got some resistant plants.As we know that, both the quantity and quality of Fructus Amomi diminishing, suffering serious diseases. Application of gene engineering is very significant and beneficial to modernization and sustaining development of Chinese material medica, which can be used in inhancing antimicrobial ability and increasing content of active composition, even in construction of transgenic cells to produce high value composition of Chinese material medica.In this dissertation, studying on effects of different explants and different plant growth substances on callus induction in AVL were investigated by Orthogonal design firstly. To establish transgenic receptor system for callus of Amomum villosum Lour.. Then Using callus of Amomum villosum Lour. as the gene transformation acceptors, the EHA105/pCAMBIA1305.2-CN gene was transferred into callus with Agrobacterium tumefaciens-mediated transformation. The factors which affected transformation were investigated by Orthogonal design. To initially establish the agrobacterium-mediated transformation Amomum villosum system of foreign Cecropin gene with the purpose of modernization and sustaining development of Chinese material medica. 1 To establish transgenic receptor system for callus of Amomum villosum Lour. studying on effects of different explants and different plant growth substances on callus induction in AVL were investigated by Orthogonal design firstly. To establish transgenic receptor system for callus of Amomum villosum Lour.. Results show that, the best optimum medium is MS added 0.5 mg·L-1 2,4-D,2 mg·L-1 6-BA,6mg·L-1AgNO3, lmg·L-1 Acid hydrolysis casein. Immaturate AVL seeds is more suitable for callus induction in Amomum villosum Lour. than mature AVL seeds., the highest rate of callus induction was 65.41%. callus in differentialmedium had got adventitious root. culture medium with 5 mg·L-1Hyg,500mg·L-1 Km are suitable for selection of resistant callus.2 Agrobacterium tumefaciens-mediated transformation of antimicrobial peptide gene CN for callus of Amomurn villosum Lour.Using callus of Amomum villosum Lour. as the gene transformation acceptors, the CN gene was transferred into callus with Agrobacterium tumefaciens-mediated transformation. The factors which affected transformation were investigated by Orthogonal design. The results showed that, using transient gus expression as a criteria, Agrobacterium tumefaciens instrusion with OD600=0.5 in 10 min, co-culture for 4 days in 24℃, AS at 200μmol·L-1 produced the highest transformation efficiency. PCR screening showed that the callus with blue dots were positive. The way by Agrobacterium tumefaciens transfection was good for genetic transformation of Amomum villosum Lour. with high transformation efficiency.In this dissertation, to establish transgenic receptor system for callus of Amomum villosum Lour. and CN gene was transferred into callus with Agrobacterium tumefaciens-mediated transformation to initially establish the agrobacterium-mediated transformation Amomum villosum system. This paper lay a foundation for genetic transformation in Amomum villosum.
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