State Key Laboratory of Food Science and Technology, Jiangnan University, Wuxi, Jiangsu, P.R. China.
School of Food Science and Technology, Jiangnan University, Wuxi, Jiangsu, P.R. China.
Lett Appl Microbiol. 2020 May;70(5):388-393. doi: 10.1111/lam.13286. Epub 2020 Mar 9.
Four Agrobacterium tumefaciens strains AGL-1, C58C1, EHA105 and LBA4404 were tested for the effects of strain types on the transformation efficiency in Mortierella alpina. Results showed that AGL-1, C58C1 and EHA105 transformed M. alpina successfully. Among them, A. tumefaciens EHA105 was first proven successful transformation of M. alpina. AGL-1 and EHA105 had the highest transformation efficiency among the four strains, while LBA4404 failed to transform M. alpina. The reason leading to the transformation efficiency difference among the four strains was explored by determining transcription levels of the virulence (vir) gene in the induction medium. Results showed that the expressions of virD1, virD2, virD4 and virE1 genes were obviously induced by acetosyringone in all the strains, and their transcriptional levels as well as virA's of AGL-1, C58C1 and EHA105 were higher than that of LBA4404, suggesting high transcriptional levels of vir genes were important for successful transformation. The study selected A. tumefaciens with high transformation efficiency of M. alpina, and would accelerate the genetic management of M. alpina. SIGNIFICANCE AND IMPACT OF THE STUDY: Oleaginous filamentous fungus Mortierella alpina is a commercial strain for the production of arachidonic acid. Genetic manipulation of M. alpina requires highly efficient transformation method. In this study, we explore the effect of Agrobacterium tumefaciens strain types on the transformation efficiency of M. alpina and select A. tumefaciens with the highest transformation efficiency, which accelerates the genetic manipulation of M. alpina. Besides, high transcriptional levels of virulence genes in A. tumefaciens were proven to play an important role for successful transformation.
四种根癌农杆菌菌株 AGL-1、C58C1、EHA105 和 LBA4404 被测试用于研究菌株类型对高山被孢霉转化效率的影响。结果表明，AGL-1、C58C1 和 EHA105 成功转化了高山被孢霉。其中，根癌农杆菌 EHA105 首次被证明成功转化了高山被孢霉。AGL-1 和 EHA105 这两种菌株在这四种菌株中的转化效率最高，而 LBA4404 未能转化高山被孢霉。通过在诱导培养基中测定毒性（vir）基因的转录水平，探讨了导致这四种菌株转化效率差异的原因。结果表明，所有菌株中 virD1、virD2、virD4 和 virE1 基因的表达均被乙酰丁香酮明显诱导，其转录水平以及 AGL-1、C58C1 和 EHA105 的 virA 高于 LBA4404，表明毒性基因的高转录水平对于成功转化是重要的。本研究选择了对高山被孢霉具有高效转化效率的根癌农杆菌，将加速高山被孢霉的遗传管理。研究的意义和影响：产ARA 的油脂丝状真菌高山被孢霉是一种商业生产 arachidonic acid 的菌株。高山被孢霉的遗传操作需要高效的转化方法。在本研究中，我们探讨了根癌农杆菌菌株类型对高山被孢霉转化效率的影响，并选择了转化效率最高的根癌农杆菌，这加速了高山被孢霉的遗传操作。此外，根癌农杆菌中毒性基因的高转录水平被证明对成功转化起着重要作用。
