Novel specific primers for rapid identification of Macrophomina species

Santos, Kledson Mendes dos; Lima, Graziele S.; Barros, Ana Paula; Machado, Alexandre Reis; Souza-Motta, Cristina Maria de; Correia, Kamila C.; Michereff, Sami Jorge

doi:10.1007/s10658-020-01952-8

Cited by 34 publications

(20 citation statements)

References 13 publications

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“…Species-specific oligonucleotide primers and oligonucleotide probes can be used to rapidly detect and identify M. phaseolina by polymerase chain reaction (PCR) and hybridization (Babu et al, 2007). More recently, specific primers have been developed for the identification of M. phaseolina, M. pseudophaseolina, and M. euphorbiicola (Santos et al, 2020). This may contribute to broader studies conducted to evaluate the diversity and distribution of species of this genus.…”

Section: Molecular Diagnosticsmentioning

confidence: 99%

Macrophomina phaseolina: General Characteristics of Pathogenicity and Methods of Control

et al. 2021

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Macrophomina phaseolina is a generalist soil-borne fungus present all over the world. It cause diseases such as stem and root rot, charcoal rot and seedling blight. Under high temperatures and low soil moisture, this fungus can cause substantial yield losses in crops such as soybean, sorghum and groundnut. The wide host range and high persistence of M. phaseolina in soil as microsclerotia make disease control challenging. Therefore, understanding the basis of the pathogenicity mechanisms as well as its interactions with host plants is crucial for controlling the pathogen. In this work, we aim to describe the general characteristics and pathogenicity mechanisms of M. phaseolina, as well as the hosts defense response. We also review the current methods and most promising forecoming ones to reach a responsible control of the pathogen, with minimal impacts to the environment and natural resources.

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Section: Molecular Diagnosticsmentioning

confidence: 99%

Macrophomina phaseolina: General Characteristics of Pathogenicity and Methods of Control

et al. 2021

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“…On a molecular level, RPB2 sequences are recommended for accurate molecular identi cation of Ramulariopsis isolates, given its universal application, speed, and the presumption that this molecular marker safely approximates taxonomic expertise. However, this technique is laborious, expensive, and requires time and knowledge of phylogenetic analysis for identifying the species 27 . The desire for rapid, automated approaches, such as that obtained here using the ISSR (TGTC) 4 primer, indicates that the RPB2 region can potentially be used for easy and inexpensive diagnosis and detection assays.…”

Section: Discussionmentioning

confidence: 99%

Ramularia leaf spot: PCR-based methods reveal widespread distribution of Ramulariopsis pseudogycines and limited presence of R. gossypii in Brazil

Silva¹,

Rennó²,

Quitania³

et al. 2021

Preprint

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Whilst Brazil is the fourth largest cotton producer globally, incidence of ramularia leaf spot (RLS) has decreased yield across all growing regions. In 2017-18 and 2018-19 growing seasons, ca. 300 fungal samples were collected from seven Brazilian states. Hyphal tip cultures were obtained for DNA extraction and amplification of partial sequences of the RNA polymerase II gene (RPB2), 28S rRNA, the ribosomal DNA internal transcribed spacers (ITS), actin (ACT), elongation factor (EF1-α) and histone H3 (HIS3) gene regions. Genetic diversity of the isolates was evaluated using fourteen molecular markers. Clade assignments based on the concatenated-sequence tree (RPB2, LSU, EF1-α, ITS, ACT, and HIS3) were identical to those in tree clades generated by RPB2-sequences, as well as in an RPB2 haplotype network, in an ISSR (TGTC)4 dendrogram, and based on morphological comparisons. In total, 252 out of 267 isolates were identified as Ramulariopsis pseudoglycines, indicating this species as the most widespread causal agent of cotton RLS in the Brazilian growing regions today. Validation of the ISSR (TGTC)4 primer as a tool to study the diversity and distribution of Ramulariopsis species will make it possible to carry out extensive RLS sampling studies worldwide.

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“…The genomic DNA of the isolate was extracted using cetyl trimethylammonium bromide (CTAB) method [12]. The primer Btub‐F1/Btub‐R1 was used to amplify the β‐tubulin gene [13] and ITS1/ITS4 primers were used to amplify the sequence of internal transcriptional spacer (ITS) of ribosomes [14]. Primer Yph1F/Yph2R amplified the encoding gene of Ypt1 associated with the Ras family of Phytophthora [15].…”

Section: Methodsmentioning

confidence: 99%

Phytophthora cinnamomi causing root rot on Rhododendron lapponicum and control it using potential biocontrol agents

Liu

Munir

et al. 2022

J Basic Microbiol

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Rhododendron lapponicum (R. lapponicum) is a dwarf Rhododendron species, which is severely infected with root rot and wilt in Yunnan province, China. However, the causal agent causing these symptoms was unknown. An isolate, Pci‐1 was identified as Phytophthora cinnamomi, based on its morphology and the sequences of β‐tubulin, internal transcribed spacer, and Ypt1 genes and verified according to the Koch's postulate. We found that this pathogen could infect 14 species of plants, including Althaea rosea, Viburnum cylindricum, and Brassica napus. Strain Pci‐1 could cause R. lapponicum to wither and die; and it grows best in an oat medium with pH 7.0 − 8.0 and an optimum temperature of 27°C. We suggest that the rhizosphere of R. lapponicum treated with biocontrol strains Paenibacillus polymyxoides P2‐5 and Trichoderma asperellum Tv‐1 showed a significant inhibitory effect on pathogen Pci‐1. The inhibitory effect of 70% dimethomorph + cymoxanil was significantly higher with EC50 and EC90 values of 0.1894 and 0.3618 a.i. µg/ml, respectively. Greenhouse experiments revealed that the pathogen load is decreased in the presence of potential antagonists. This study provides fundamentals on risk assessment and theoretical support for the management of P. cinnamomi pathogen and contributes significantly to the planting of forest and horticultural crops in a disease‐free environment.

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Novel specific primers for rapid identification of Macrophomina species

Cited by 34 publications

References 13 publications

Macrophomina phaseolina: General Characteristics of Pathogenicity and Methods of Control

Macrophomina phaseolina: General Characteristics of Pathogenicity and Methods of Control

Ramularia leaf spot: PCR-based methods reveal widespread distribution of Ramulariopsis pseudogycines and limited presence of R. gossypii in Brazil

Phytophthora cinnamomi causing root rot on Rhododendron lapponicum and control it using potential biocontrol agents

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