Efficient Agrobacterium tumefaciens-mediated transformation system of Diaporthe caulivora

Montoya, Marina Rosa; Massa, G; Colabelli, Mabel Noemí; Ridao, Azucena

doi:10.1016/j.mimet.2021.106197

Cited by 2 publications

(3 citation statements)

References 61 publications

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“…As shown in Figure 1D, the yield of transformants was highest after 48 h of co-cultivation and was >30 times higher than that after 24 h of co-cultivation. Such a short co-cultivation period results in a lower number of transformation events compared to those after 36, 48, or 72 h [13,15,28,29,37,42,43].…”

Section: Discussionmentioning

confidence: 99%

Agrobacterium tumefaciens-Mediated Transformation of the Aquatic Fungus Phialemonium inflatum FBCC-F1546

Yoon,

Kim,

Kim

et al. 2023

JoF

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Phialemonium inflatum is a useful fungus known for its ability to mineralise lignin during primary metabolism and decompose polycyclic aromatic hydrocarbons (PAHs). However, no functional genetic analysis techniques have been developed yet for this fungus, specifically in terms of transformation. In this study, we applied an Agrobacterium tumefaciens-mediated transformation (ATMT) system to P. inflatum for a functional gene analysis. We generated 3689 transformants using the binary vector pSK1044, which carried either the hygromycin B phosphotransferase (hph) gene or the enhanced green fluorescent protein (eGFP) gene to label the transformants. A Southern blot analysis showed that the probability of a single copy of T-DNA insertion was approximately 50% when the co-cultivation of fungal spores and Agrobacterium tumefaciens cells was performed at 24–36 h, whereas at 48 h, it was approximately 35.5%. Therefore, when performing gene knockout using the ATMT system, the co-cultivation time was reduced to ≤36 h. The resulting transformants were mitotically stable, and a PCR analysis confirmed the genes’ integration into the transformant genome. Additionally, hph and eGFP gene expressions were confirmed via PCR amplification and fluorescence microscopy. This optimised transformation system will enable functional gene analyses to study genes of interest in P. inflatum.

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Section: Discussionmentioning

confidence: 99%

Agrobacterium tumefaciens-Mediated Transformation of the Aquatic Fungus Phialemonium inflatum FBCC-F1546

Yoon,

Kim,

Kim

et al. 2023

JoF

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“…caulivora and D . longicolla (Montoya et al., 2021; Zhang et al., 2023), together with genomic analysis, allows the identification and functional validation of fungal genes associated with pathogenicity. This will generate a more comprehensive understanding of the pathogen infection strategies and the molecular mechanisms employed to counteract the plant immune system.…”

Section: Soybean Stem Canker Managementmentioning

confidence: 99%

“…The availability of efficient transformation methods for D. caulivora and D. longicolla (Montoya et al, 2021;Zhang et al, 2023), together with genomic analysis, allows the identification and functional validation of fungal genes associated with pathogenicity.…”

Section: Host Resistancementioning

confidence: 99%

Current understanding of the Diaporthe/Phomopsis complex causing soybean stem canker: A focus on molecular aspects of the interaction

Mena,

Stewart,

Montesano

et al. 2023

Plant Pathology

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Soybean stem canker (SSC) is an important disease caused by different Diaporthe spp., including D. aspalathi, D. caulivora and D. longicolla, that leads to soybean (Glycine max) yield losses around the world. Most studies have been focused on the morphological characterization and molecular identification of Diaporthe spp. present in SSC lesions. Several soybean resistance loci to Diaporthe spp. causing SSC have been identified, although the molecular identities of the resistance genes are at present unknown. In this review, we summarize the current knowledge on SSC disease, the molecular characterization of Diaporthe spp. and their evolutionary relationships. We highlight how recent genomic and transcriptomic information is allowing significant progress in our understanding of the molecular components and mechanisms underlying Diaporthe infection strategies as well as soybean disease resistance. The information generated, combined with available resources enabling functional genomics, will contribute to the development of breeding strategies for disease resistance, leading to a more sustainable agriculture.

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Efficient Agrobacterium tumefaciens-mediated transformation system of Diaporthe caulivora

Cited by 2 publications

References 61 publications

Agrobacterium tumefaciens-Mediated Transformation of the Aquatic Fungus Phialemonium inflatum FBCC-F1546

Agrobacterium tumefaciens-Mediated Transformation of the Aquatic Fungus Phialemonium inflatum FBCC-F1546

Current understanding of the Diaporthe/Phomopsis complex causing soybean stem canker: A focus on molecular aspects of the interaction

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