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    秦爱光, 罗晓芳. 农杆菌介导转录因子DREB1C基因转化速生型刺槐的研究[J]. 北京林业大学学报, 2007, 29(6): 29-34. DOI: 10.13332/j.1000-1522.2007.06.011
    引用本文: 秦爱光, 罗晓芳. 农杆菌介导转录因子DREB1C基因转化速生型刺槐的研究[J]. 北京林业大学学报, 2007, 29(6): 29-34. DOI: 10.13332/j.1000-1522.2007.06.011
    QIN Ai-guang, LUO Xiao-fang. Transformation of transcription factor DREB1C gene into the fast-growing black locust mediated with Agrobacterium tumefaciens[J]. Journal of Beijing Forestry University, 2007, 29(6): 29-34. DOI: 10.13332/j.1000-1522.2007.06.011
    Citation: QIN Ai-guang, LUO Xiao-fang. Transformation of transcription factor DREB1C gene into the fast-growing black locust mediated with Agrobacterium tumefaciens[J]. Journal of Beijing Forestry University, 2007, 29(6): 29-34. DOI: 10.13332/j.1000-1522.2007.06.011

    农杆菌介导转录因子DREB1C基因转化速生型刺槐的研究

    Transformation of transcription factor DREB1C gene into the fast-growing black locust mediated with Agrobacterium tumefaciens

    • 摘要: 引自匈牙利的刺槐是可用于观赏、水土保持、饲料和蜂源的速生型树种,向这个树种转入耐逆基因,可满足干旱、盐碱地植被恢复的需求.该文对速生型刺槐进行了3个方面的研究:在含有不同组合6-BA和NAA的MS培养基上的芽再生情况、根癌农杆菌介导GUS(载体为pCAMBIA 1301)基因转化的优化以及耐逆转录因子基因AtDREB1C(相同载体)的转化.结果表明:①以无菌苗复叶叶轴为外植体,在附加6-BA 1.0 mg/L和NAA 0.5 mg/L的MS培养基上芽再生率最高,为77.5%;②同样的外植体在MS培养基上预培养0~2 d后,用附加乙酰丁香酮的根癌农杆菌菌液侵染15~20 min,再接种于附加20 mg/L乙酰丁香酮和Hyg 7 mg/L的上述MS培养基上,共培养2 d,可获得最佳转化效果;③用上述优化的方法将AtDREB1C导入刺槐,获得转基因植株,并得到PCR、PCR-Southern blotting和Southern blotting的验证.

       

      Abstract: Robinia pseudocacia,introduced from Hungary,is a fast-growing variety used in ornamental,soil and water conservation,fodder and nectar sources.Transformation of this woody plant using abiotic-stress-tolerant gene could meet the requirements for reclamation of arid or alkalid lands and vegetation restoration.In this paper,three aspects of R.pseudocacia were studied: shoot regeneration on MS medium with different combinations of 6-BA and NAA,optimization of β-glucuronidase(GUS) gene(carried by vector pCAMBIA 1301,and mediated by Agrobacterium tumefaciens) transformation,and gene transformation using the transcription factor AtDREB1C gene (abiotic-stress-tolerant gene carried by the same vector).Results showed that: 1) The highest shoot-regeneration rate 77.5% was obtained from the rachis of compound leaves cultured on MS base medium combimed with 6-BA 1.0 mg/L and NAA 0.5 mg/L;2) The best transformation efficiency was proved by the rachis of compound leaves proculturing for 0—2 days on the MS medium,immerging into the solution containing Agrobacterium and acetosyingone(AS) for 15-20 min,subsequencely co-culturing for two days on the same medium with AS 20 mg/L and hygromycin(Hyg) 7 mg/L;3) Following the method optimized,the AtDREB1C gene was introduced into R.pseudocacia,and transgenic plants were obtained and verified by PCR,PCR-Southern blotting and Southern blotting analysis.

       

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