A newly constructed Agrobacterium-mediated transformation system based on the hisB auxotrophic marker for genetic manipulation in Aspergillus niger

Thai, Hanh-Dung; Xuan, Loc Thi Binh; Nguyen, Huan X.; Vu, Thang X.; Tran, Huyen Thi Thanh; Nguyen, Huy Quang; Tran, Van‐Tuan

doi:10.1007/s00203-023-03530-y

Cited by 4 publications

(1 citation statement)

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“…Meanwhile, the Agrobacterium tumefaciens -mediated transformation (ATMT) system is gaining popularity as an effective insertional mutagenesis method for gene overexpression, gene targeting, and T-DNA insertional mutagenesis in a variety of filamentous fungus [ 24 , 25 , 26 ]. In contrast to protoplast-mediated transformation (PMT), ATMT confers a discernible advantage, given its capacity to facilitate the direct exploitation of fungal spores for genetic manipulation, obviating the requisite for protoplasts [ 27 ]. Despite its widespread application in various filamentous fungi, the successful implementation of ATMT within the context of L. qinlingensis has yet to be realized.…”

Section: Introductionmentioning

confidence: 99%

Establishment of RNA Interference Genetic Transformation System and Functional Analysis of FlbA Gene in Leptographium qinlingensis

Gan,

An,

Tang

et al. 2023

IJMS

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Leptographium qinlingensis is a pathogenic fungus of Pinus armandii that is epidemic in the Qinling Mountains. However, an effective gene interference strategy is needed to characterize the pathogenic genes in this fungus on a functional level. Using the RNA silencing vector pSilent-1 as a template, we established an RNA interference genetic transformation system mediated by Agrobacterium tumefaciens GV3101, which is suitable for the gene study for Leptographium qinlingensis by homologous recombination and strain interference system screening. The LqFlbA gene was silenced using the RNA interference approach described above, and the resulting transformants displayed various levels of silencing with a gene silencing effectiveness ranging from 41.8% to 91.4%. The LqFlbA-RNAi mutant displayed altered colony morphology, sluggish mycelium growth, and diminished pathogenicity toward the host P. armandii in comparison to the wild type. The results indicate that this method provides a useful reverse genetic system for studying the gene function of L. qinlingensis, and that LqFlbA plays a crucial role in the growth, development, and pathogenicity of L. qinlingensis.

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