LI Jianyong.Transformation Techniques for Petunia hybrida With ACC Deaminase Gene Driven by Different Promoters[J].Northern Horticulture,2016,40(20):85-91.[doi:10.11937/bfyy.201620023]
不同启动子驱动的ACDS基因对矮牵牛遗传转化技术研究
- Title:
- Transformation Techniques for Petunia hybrida With ACC Deaminase Gene Driven by Different Promoters
- Keywords:
- Petunia hybrida; promoter; ACC deaminase gene; genetic transformation
- 文献标志码:
- A
- 摘要:
- 以矮牵牛为试材，通过根癌农杆菌介导ACC脱氨酶基因(ACDS)（分别由CaMV35S、SAG12和CHSA启动子驱动）转化矮牵牛叶盘，探讨了预培养时间、农杆菌菌液浓度、浸染时间、共培养时间、乙酰丁香酮(AS)浓度等因素对转化的影响，比较了ACDS基因在不同启动子驱动下获得转基因植株的效率及转基因植株的生长速率。结果表明：预培养5 d的矮牵牛叶盘在含AS 200 μmol?L-1、OD600值为0.6的农杆菌菌液中浸染8 min后，共培养4 d时遗传转化效果最优；不同启动子驱动的ACDS基因转化效率以CHSA启动子效率最高(3%），SAG12启动子效率最低(1.25%)。PCR检测证明外源基因已整合进入矮牵牛基因组中。转ACDS基因矮牵牛植株的获得，为利用基因工程技术培育抗衰老矮牵牛新品种奠定了技术基础。
- Abstract:
- Taking Petunia hybrida as material，ACC deaminase gene(ACDS) (driven respectively by promoter CaMV35S，SAG12 and CHSA) was transformed into Petunia hybrida leaves，mediated by Agrobacterium tumefaciens.The effects of the pre-culture time，concentration of Agrobacterium tumefaciens，infection time，co-culture time and concentration of AS on transformation were studied.The transformation efficiency of ACDS gene driven by different promoters was compared based on the percentage of transgenic plants and the growth rate of transgenic plants.The results showed that the best transformation efficiency was abtained under the conditions of Petunia hybrida leaves pre-culture for 5 days，infection for 8 minutes in Agrobacterium tumefaciens liquid of 0.6 OD600 value containing 200 μmol?L-1 acetosyringone，and then co-culture for 4 days.ACDS gene driven by CHSA promoter had the highest transformation efficiency(3%），while ACDS gene driven by SAG12 promoter had the lowest transformation efficiency(1.25%).PCR analysis verified that the foreign gene was integrated into the Petunia hybrida genome.The obtaining of Petunia hybrida transgenic plants set up a technical basis for breeding of anti-aging Petunia hybrida varieties via genetic engineering.
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备注/Memo
作者简介：李建永（1981-），男，硕士，讲师，研究方向为植物遗传育种与基因工程。E-mail：ljy_518@163.com.基金项目：国家自然科学基金资助项目（30560095）。