Rapid and sensitive detection of <i>Curvularia lunata</i>
 associated with maize leaf spot based on its <i>Clg2p</i>
 gene using semi-nested PCR

Hou, Jumei; Ma, B.C.; Zuo, Yi; Guo, Li; Gao, Shigang; Wang, Y.Y.; Liu, T.

doi:10.1111/lam.12040

Cited by 17 publications

(14 citation statements)

References 20 publications

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“…The morphological characterization of C. lunata was verified through observations of the macroscopic and microscopic characteristics according to previous studies (Ferreira et al, 2010;Kern and Blevins, 1999). For the molecular characterization done according to Hou et al (2013), DNA extraction was performed using a cetyl trimetyl ammonium bromide (CTAB) method described by Zolan and Pukilla (1986). The primers used were P1: (5ʹ-ATG GAC GAG AAC AAC AGG ATAA CGA-3ʹ) and P2: (5ʹ-CTA CCA GCA TTT GAA TTT ACT CCAG-3ʹ).…”

Section: Morphological and Molecular Characterization Of C Lunatamentioning

confidence: 99%

Fungistatic activity of essential oil of Lippia sidoides Cham. against Curvularia lunata

Ferreira¹,

MourÃ£o²,

Santos³

et al. 2018

Afr. J. Agric. Res.

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Synthetic pesticides used in agriculture to control pathogens are being widely questioned for their harmful effects on human health and the environment. Therefore, the objective of this study was to evaluate the fungistatic activity of Lippia sidoides Cham. essential oil on alternative control of Curvularia lunata (Wakker), the causal agent of spot in maize plants. The fungitoxic contact effect was successful, resulting in a strong inhibition of C. lunata at the concentration of 50 mg mL −1. Complete adverse effect on the germination of the conidia of C. lunata was obtained at concentrations of 5 and 7.5 mg mL −1. In the preventive assay in maize plants, that is, effect of essential oil on C. lunata, a reduction in the progression of the disease was observed at the oil concentration of 7.5 mg mL −1. Regarding the curative effect in maize plants, no satisfactory result was obtained at concentrations from 0.625 to 7.5 mg mL −1. Cytotoxic test in animal cells was also performed by the viability test using MTT assay, which showed that none of the analyzed concentrations was toxic to the cells. These results demonstrate the potential effect of the essential oil of L. sidoides on the prevention and inhibition of mycelia and conidia germination, controlling the phytopathogenic fungus C. lunata.

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Section: Morphological and Molecular Characterization Of C Lunatamentioning

confidence: 99%

Fungistatic activity of essential oil of Lippia sidoides Cham. against Curvularia lunata

Ferreira¹,

MourÃ£o²,

Santos³

et al. 2018

Afr. J. Agric. Res.

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“…Since the Ferreira et al 297 1990s, it has caused economic loss in maize crop productivity in China (Gao et al, 2015) and has been progressing in recent years in Brazil (Assunção et al, 2006). The pathogen mainly infects maize leaves, leaf sheaths, and husks and causes soaked or yellow necrotic spots in the early stages, which then expand to round, oval, spindle-shaped, or strip lesions (Hou et al, 2013). The green leaf area of maize is considered the main source of photoassimilates for the plant (Akram et al, 2014) and, according to Liu et al (2016), a loss in this source may reflect the development of the plant and the production of grains.…”

Section: Introductionmentioning

confidence: 99%

Secondary metabolites from endophytic fungus from Lippia sidoides Cham.

Ferreira¹,

Santos²,

Soares³

et al. 2017

J. Med. Plants Res.

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Lippia sidoidesCham. (Verbenaceae) is a species native to the Brazilian northeast, widely used in popular medicine. Its leaves were used for the isolation of endophytic fungi and extraction of metabolites. Among them, three were selected according to fungitoxicity tests against the maize phytopathogenic fungus, Curvularia lunata (Wakker). However, the objective of this study was to identify the role of L. sidoides extracts associated with their endophytic fungi, necessary to reduce excess of fungicides applied on the maize crop. Metabolites were evaluated for antioxidant activity by 2.2-diphenyl-1-picrylhydrazyl (DPPH), phenols, total flavonoids and one of it endophytic fungus were evaluated for synergism (Verticilium sp. and plant extracts). The endophytic fungi and plant extracts evaluated for phenolic content ranged from 0.29 ± 0.05 to 96.94 ± 11.86 mgEAG/g, the content of flavonoids from 14.31 ± 1.56 to 192.33 ± 4.58 mgER/g, and antioxidant activity could only be observed for the plant extract with EC 50 81 ± 0.3%. The secondary metabolites identified by HPLC in the plant extract were catechin, quercetin, gallic acid and naringin. Naringenin, catechin, epigallocatechin gallate and quercetin were identified in the extract of the fungi viz. Verticillium sp. and Fusarium sp. Synergistic analysis between a 1:1 proportion of plant and fungal extracts has shown more efficient (79.0%) inhibition of C. lunata. Thus, alternative control of phytopathogenic fungi can be accomplished using plant extracts associated with their endophytic fungi, reducing the excess of fungicides applied on the maize crop.

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“…The development of accurate and reliable methods for fungus identification has increased; most of these methods are based on sensitive PCR to enable the use of specific genetic markers (Hou et al, 2013). DNA extraction is an important initial step of genetic identification of fungi.…”

Section: Discussionmentioning

confidence: 99%

An economical and combined method for rapid and efficient isolation of fungal DNA

2014

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DNA isolation is a crucial step of conducting genetic studies in any organism. However, this process is quite difficult when studying fungi because of the need to damage the fungal cell walls of specific structures. In this study, we developed a method for the rapid and efficient isolation of fungal DNA based on simultaneous mechanical and enzymatic cell wall degradation. There are several typical modifications of the standard phenol-chloroform DNA extraction method. This method can be modified to degrade the fungal cell wall. The first step of the presented DNA extraction included manual homogenization in modified lysis buffer. Next, enzymatic digestion using 2 enzymes was conducted, including lyticase and proteinase K. To carefully select the most favorable conditions, we developed an economical, rapid, and reliable method for fungal DNA extraction that ensures both high efficiency and proper purity, which are essential for further analyses.

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Rapid and sensitive detection of Curvularia lunata associated with maize leaf spot based on its Clg2p gene using semi-nested PCR

Cited by 17 publications

References 20 publications

Fungistatic activity of essential oil of Lippia sidoides Cham. against Curvularia lunata

Fungistatic activity of essential oil of Lippia sidoides Cham. against Curvularia lunata

Secondary metabolites from endophytic fungus from Lippia sidoides Cham.

An economical and combined method for rapid and efficient isolation of fungal DNA

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