农杆菌介导的ACO反义基因对铁皮石斛的遗传转化  

Genetic Transformation of ACO Antisense Gene into Dendrobium officinale Kimura et Migo. by Agrobacterium Mediation

作　　者：武荣花[1] 康莹莹[1] 王洁琼[2] 刘佳[2] 崔波[2,3] 蒋素华[3] 张开明[1] 

机构地区：[1]河南农业大学林学院,河南郑州450002 [2]河南农业大学生命科学学院,河南郑州450002 [3]郑州师范学院生物工程研究所,河南郑州450044

出　　处：《华北农学报》2015年第2期17-21,共5页Acta Agriculturae Boreali-Sinica

基　　金：国家自然科学基金项目(31101562);河南省高等学校重点科研项目(15A220006)

摘　　要：以铁皮石斛原球茎为受体,用农杆菌介导1-氨基环丙烷-1-羧酸氧化酶(ACO)的反义基因对铁皮石斛进行遗传转化,并对影响转化的几个因素进行优化,结果表明:经刀切法处理的原球茎,在AS浓度为100μmol/L、农杆菌菌液OD600值为0.8的条件下侵染30 min,共培养60 h,接种到含有30 mg/L Mer和3.0 mg/L PPT的筛选培养基中,遗传转化效率最高。筛选获得的抗性植株经GUS组织化学染色和PCR鉴定,初步证明ACO反义基因已整合到铁皮石斛的基因组中。The ACO antisense gene was transformed into the protocorms of Dendrobiun officinale by Agrobacte-rium mediation. To optimize the transformation efficiency,several related factors were studied in this paper. The re-sults showed that,after infected for 30 min at 100 μmol/L of AS,0. 8 of OD600 and 60 h of incubation in darkness, the cut protocorms gained the highest transformation rate in the medium of 30 mg/L Mer and 3. 0 mg/L PPT. The selected plants were tested by PCR and GUS histochemical staining,indicating that the ACO antisense gene had been successfully integrated into D. officinale.

关 键 词：铁皮石斛 ACC氧化酶 遗传转化 根癌农杆菌 

分 类 号：Q78[生物学—分子生物学]