Development qRT-PCR Protocol to Predict Strawberry Fusarium Wilt Occurrence

Hong, Sung Won; Kim, Da-Ran; Kim, Ji Su; Cho, Gyeongjun; Jeon, Chang Wook; Kwak, Youn‐Sig

doi:10.5423/ppj.oa.12.2017.0265

Cited by 5 publications

(1 citation statement)

References 33 publications

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“…Currently, qRT‐PCR is one of the most important tools in genetic and molecular research, as it can quantify gene expression levels accurately and efficiently (Hong et al 2018). Because endogenous reference genes can standardise the qRT‐PCR results by reducing inevitable experimental errors, obtaining perfect qRT‐PCR results greatly depends on the selection of reference genes (Dheda et al 2005, Wang et al 2014, 2019a).…”

Section: Discussionmentioning

confidence: 99%

Identification of optimal and novel reference genes for quantitative real‐time polymerase chain reaction analysis in grapevine

Wei

Wang

Liu

et al. 2021

Australian Journal of Grape and Wine Research

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Background and Aims Quantitative real‐time polymerase chain reaction (qRT‐PCR) has the advantages of sensitivity, specificity and repeatability, and is commonly used to investigate the expression levels of genes. The selection of appropriate endogenous reference genes is the pre‐condition for acquiring the desired results of qRT‐PCR. This study aimed to identify and screen the most suitable reference genes for grapevines. Methods and Results Three novel genes from previous transcriptomic data were selected for their stable expression either at the developmental stages of the grape berry or under the various treatments. The expression stability of these three candidate genes and six commonly used reference genes were evaluated with geNorm, NormFinder and BestKeeper algorithms in several grapevine samples. The geNorm analysis indicated that two reference genes were sufficient to normalise expression. The optimal reference gene combinations were: VvEF1‐γ and PPR2 in fruit; RRM1 and EF1‐α in leaf; EF1‐α and Actin in tendril of several cultivars. The combination of EF1‐α and VvEF1‐α was the most appropriate reference gene for fruits at different developmental stages. When all the samples were considered, the combination of RRM1 and Ubiquitin was optimal. Conclusions Different reference genes should be employed when qRT‐PCR is performed in different grapevine samples. The gene RRM1 is a new appropriate reference gene for grapevine. Significance of the Study This study provided alternatives for the optimal reference genes in qRT‐PCR, and identified one novel and appropriate reference gene (RRM1) for grapevine.

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Section: Discussionmentioning

confidence: 99%

Identification of optimal and novel reference genes for quantitative real‐time polymerase chain reaction analysis in grapevine

Wei

Wang

Liu

et al. 2021

Australian Journal of Grape and Wine Research

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Function and Distribution of a Lantipeptide in Strawberry Fusarium Wilt Disease–Suppressive Soils

Kim

Jeon

Shin

et al. 2019

MPMI

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Streptomyces griseus S4-7 is representative of strains responsible for the specific soil suppressiveness of Fusarium wilt of strawberry caused by Fusarium oxysporum f. sp. fragariae. Members of the genus Streptomyces secrete diverse secondary metabolites including lantipeptides, heat-stable lanthionine-containing compounds that can exhibit antibiotic activity. In this study, a class II lantipeptide provisionally named grisin, of previously unknown biological function, was shown to inhibit F. oxysporum. The inhibitory activity of grisin distinguishes it from other class II lantipeptides from Streptomyces spp. Results of quantitative reverse transcription-polymerase chain reaction with lanM-specific primers showed that the density of grisin-producing Streptomyces spp. in the rhizosphere of strawberry was positively correlated with the number of years of monoculture and a minimum of seven years was required for development of specific soil suppressiveness to Fusarium wilt disease. We suggest that lanM can be used as a diagnostic marker of whether a soil is conducive or suppressive to the disease.

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Detection of Fusarium oxysporum f. sp. fragariae by Using Loop-Mediated Isothermal Amplification

Katoh

Yamazaki²,

Fukuda³

et al. 2021

Plant Disease

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We developed a loop-mediated isothermal amplification (LAMP) assay for detecting Fusarium oxysporum f. sp. fragariae, the causal agent of wilt in strawberry plants. This assay was based on genomic regions between the portions of transposable elements Han and Skippy of the fungus. The LAMP assay allowed the efficient detection of F. oxysporum f. sp. fragariae DNA by visual inspection, without requiring gel electrophoresis. The detection limit was 100 pg of genomic DNA, which is comparable to that of polymerase chain reaction (PCR). The LAMP primers successfully discriminated F. oxysporum f. sp. fragariae strains from non-pathogenic F. oxysporum strains and other fungi. The LAMP assay at 63°C, which was found to be the optimal treatment temperature, for 1.5 h successfully detected F. oxysporum f. sp. fragariae California strains GL1270 and GL1385. When the assay was performed using a GenelyzerTM FIII portable fluorometer, these California strains were successfully detected in 1 h. The assay facilitated the detection of conidia in soil samples after they were precultured on a selective medium for F. oxysporum (FoG2) as well as latent infection in strawberry plants after preculturing. The LAMP assay for visual inspection of DNA required only a heating block and an incubator, reducing the cost of this assay. Thus, it could be suitable for the detection of F. oxysporum f. sp. fragariae strains in centers that store pre-foundation and foundation stocks of strawberry, including plant nurseries.

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Development qRT-PCR Protocol to Predict Strawberry Fusarium Wilt Occurrence

Cited by 5 publications

References 33 publications

Identification of optimal and novel reference genes for quantitative real‐time polymerase chain reaction analysis in grapevine

Identification of optimal and novel reference genes for quantitative real‐time polymerase chain reaction analysis in grapevine

Function and Distribution of a Lantipeptide in Strawberry Fusarium Wilt Disease–Suppressive Soils

Detection of Fusarium oxysporum f. sp. fragariae by Using Loop-Mediated Isothermal Amplification

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