Luo Qiyu, Chen Shu, Nian Hai, Ma Qibing, Ding Yuyao, Hao Qinwen, Wei Jiping, Patel Jinesh D, McElroy Joseph Scott, Liu Yaoguang, Chen Yong
College of Agriculture, South China Agricultural University, Guangzhou 510642, China.
College of Life Sciences, South China Agricultural University, Guangzhou 510642, China.
Int J Mol Sci. 2023 Apr 1;24(7):6629. doi: 10.3390/ijms24076629.
(goosegrass) is a problematic weed worldwide known for its multi-herbicide tolerance/resistance biotype. However, a genetic transformation method in goosegrass has not been successfully established, making a bottleneck for functional genomics studies in this species. Here, we report a successful -mediated transformation method for goosegrass. Firstly, we optimized conditions for breaking seed dormancy and increasing seed germination rate. A higher callus induction rate from germinated seeds was obtained in N6 than in MS or B5 medium. Then the optimal transformation efficiency of the reporter gene was obtained by infection with culture of OD = 0.5 for 30 min, followed by 3 days of co-cultivation with 300 μmol/L acetosyringone. Concentrations of 20 mg L kanamycin and 100 mg L timentin were used to select the transformed calli. The optimal rate of regeneration of the calli was generated by using 0.50 mg L 6-BA and 0.50 mg L KT in the culture medium. Then, using this transformation method, we overexpressed the paraquat-resistant gene into a paraquat-susceptible goosegrass biotype MZ04 and confirmed the stable inheritance of paraquat-resistance in the transgenic goosegrass lines. This approach may provide a potential mechanism for the evolution of paraquat-resistant goosegrass and a promising gene for the manipulation of paraquat-resistance plants. This study is novel and valuable in future research using similar methods for herbicide resistance.
牛筋草是一种在全球范围内都存在问题的杂草,以其对多种除草剂的耐受性/抗性生物型而闻名。然而,牛筋草的遗传转化方法尚未成功建立,这成为该物种功能基因组学研究的一个瓶颈。在此,我们报道了一种成功的牛筋草农杆菌介导的转化方法。首先,我们优化了打破种子休眠和提高种子发芽率的条件。在N6培养基中,从发芽种子获得的愈伤组织诱导率高于MS或B5培养基。然后,通过用OD = 0.5的农杆菌培养物感染30分钟,随后与300μmol/L乙酰丁香酮共培养3天,获得了报告基因的最佳转化效率。使用20mg/L卡那霉素和100mg/L替门汀的浓度来筛选转化的愈伤组织。通过在培养基中使用0.50mg/L 6-苄基腺嘌呤和0.50mg/L激动素,产生了愈伤组织的最佳再生率。然后,使用这种转化方法,我们将抗百草枯基因过表达到对百草枯敏感的牛筋草生物型MZ04中,并证实了转基因牛筋草品系中百草枯抗性的稳定遗传。这种方法可能为抗百草枯牛筋草的进化提供一种潜在机制,并为操纵抗百草枯植物提供一个有前景的基因。这项研究在未来使用类似方法进行除草剂抗性研究中具有新颖性和价值。