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Comparison of molecular protocols to detect Tomato brown rugose fruit virus in solanaceae hosts 比较检测茄科寄主番茄褐皱纹果病毒的分子方案
Pub Date : 2023-12-26 DOI: 10.18781/r.mex.fit.2023-5
E. J. Zamora-Macorra, Katia Aviña-Padilla, Rosemarie W. Hammond, D. Ochoa-Martínez
The Tomato brown rugose fruit virus (ToBRFV) has emerged as a significant threat to Solanaceae crops, including tomato and pepper. Its presence in Mexico since 2018 has raised concerns about its impact on agricultural production. Early and accurate detection of this pathogen is essential to prevent its spread and mitigate its effects. In Mexico, several molecular techniques are employed for its diagnosis, including endpoint RT-PCR, RT-qPCR, and multiplex RT-qPCR. This research aimed to assess the efficiency of different RNA extraction methods in combination with specific PCR primers for detecting ToBRFV. Among the methods tested, the CTAB-Trizol RNA extraction protocol combined with nested PCR using primers reported by Dovas et al. (2004) was identified as the most sensitive molecular method for detecting the virus. This finding highlights the importance of selecting the appropriate combination of extraction and amplification protocols to achieve optimal sensitivity and accuracy in ToBRFV detection.
番茄褐皱纹果病毒(ToBRFV)已成为番茄和辣椒等茄科作物的重大威胁。2018 年以来,它在墨西哥的出现引发了人们对其对农业生产影响的担忧。早期准确检测这种病原体对于防止其传播和减轻其影响至关重要。墨西哥采用了多种分子技术对其进行诊断,包括终点 RT-PCR、RT-qPCR 和多重 RT-qPCR。本研究旨在评估不同的 RNA 提取方法结合特定 PCR 引物检测 ToBRFV 的效率。在测试的方法中,CTAB-Trizol RNA 提取方案结合使用 Dovas 等人(2004 年)报告的引物进行巢式 PCR 被认为是检测该病毒最灵敏的分子方法。这一发现凸显了选择适当的提取和扩增方案组合以达到最佳灵敏度和准确性检测 ToBRFV 的重要性。
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引用次数: 0
Genus Orthotospovirus in Costa Rica: A Central American case 哥斯达黎加的 Orthotospovirus 属:中美洲的一个案例
Pub Date : 2023-12-26 DOI: 10.18781/r.mex.fit.2023-6
M. Montero-Astúa, Natasha Dejuk-Protti, David Bermúdez-Gómez, Elena Vásquez Céspedes, I. Sandoval-Carvajal, L. Garita-Salazar, Federico J. Albertazzi, Scott Adkins, Lisela Moreira-Carmona
The Orthotospovirus genus encompasses a range of economically significant and emerging plant viruses that affect a variety of crops globally. While the prevalence and characteristics of these phytopathogenic viruses are extensively documented in North and South America, their presence in Central America remains comparatively underexplored. This study focuses on Costa Rica, strategically positioned at the nexus of North and South America, to enhance our understanding of orthotospovirus in this region. We analyzed 295 plant samples using enzyme-linked immunosorbent assay (ELISA) to test for the presence of INSV, IYSV, TSWV, and the GRSV/TCSV serogroup. Additionally, a subset (20 samples) underwent further scrutiny through reverse transcription-polymerase chain reaction (RT-PCR) employing both universal and species-specific primers. Our ELISA results indicated the absence of TSWV and the GRSV/TCSV serogroup. However, the presence of INSV in Costa Rica was substantiated through ELISA, RT-PCR, and partial sequencing, revealing its prevalence in both open-field and greenhouse environments. Despite previous diagnostic reports suggesting the presence of TSWV in Costa Rica, our study did not detect this virus. RT-PCR analysis with degenerate primers also found no evidence of other orthotospovirus species in our samples. The identification of a dominant INSV haplotype, along with three additional variants, suggests the likelihood of at least two independent virus introductions into the region. These findings underscore the necessity for more comprehensive surveys and research on orthotospoviruses in Central America to better understand their epidemiology and impact on agriculture.
直翅目病毒属包括一系列具有重要经济意义的植物病毒和新出现的植物病毒,它们在全球范围内影响着多种作物。虽然这些植物致病病毒在北美和南美的流行情况和特征已被广泛记录,但它们在中美洲的存在情况仍相对缺乏探索。哥斯达黎加地处南北美洲交界处,具有重要的战略地位,本研究将重点放在哥斯达黎加,以加深我们对该地区直翅目病毒的了解。我们使用酶联免疫吸附法 (ELISA) 分析了 295 份植物样本,检测是否存在 INSV、IYSV、TSWV 和 GRSV/TCSV 血清群。此外,我们还利用通用引物和物种特异性引物,通过反转录聚合酶链反应(RT-PCR)对部分样本(20 份)进行了进一步检测。我们的酶联免疫吸附试验结果表明不存在 TSWV 和 GRSV/TCSV 血清群。然而,通过 ELISA、RT-PCR 和部分测序,我们证实了 INSV 在哥斯达黎加的存在,揭示了它在露地和温室环境中的普遍性。尽管之前的诊断报告表明哥斯达黎加存在 TSWV,但我们的研究并未检测到这种病毒。使用退化引物进行的 RT-PCR 分析也没有在我们的样本中发现其他直翅目病毒种类。一种主要 INSV 单倍型和另外三种变体的发现表明,该地区可能至少有两种独立的病毒传入。这些发现突出表明,有必要对中美洲的直孢子虫病毒进行更全面的调查和研究,以更好地了解其流行病学和对农业的影响。
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引用次数: 0
Use of endophytic microorganisms for the management of Tomato brown rugose fruit virus in tomato crop (Solamun lycopersicum) 利用内生微生物控制番茄作物(Solamun lycopersicum)中的番茄褐皱纹病毒
Pub Date : 2023-12-26 DOI: 10.18781/r.mex.fit.2023-1
Carlos D. Ramos-Villanueva, Guadalupe Carrillo-Benitez, E. J. Zamora-Macorra, Eduardo Santiago-Elena, Samuel Ramírez-Alarcón, Jezrael Jimenez-Vidals, Ricardo Ávila-López
The Tomato brown rugose fruit virus (ToBRFV) is one of the main pathogens affecting tomato crops in Mexico. Despite efforts to prevent its spread, it is nearly impossible due to its low transmission percentage through seeds and its high susceptibility to being transmitted through cultural practices. Therefore, alternative management strategies are being sought. This research aimed to determine the effect of endophytic microorganisms applied to the soil on tomato plants infected with ToBRFV. A tomato plant was used as an experimental unit, with 13 repetitions per treatment. The treatments on tomato plants infected with ToBRFV were Beauveria peruviencis, Trichoderma longibrachiatum, Pseudomonas sp. And water as a sick witness; a treatment of healthy plants treated with water was also included as an absolute control. The response variables were plant height, fresh weight of the aerial part and root and severity (two evaluations). Measurements were analyzed using Tukey-Kramer HSD tests for each pair. Significant differences were found Beauveria peruviencis, Trichoderma longibrachiatum, Pseudomonas sp. and water as a sick witness. The treatment that most favored the development of infected plants (79% taller and 15% heavier than infected mock) and reduced its severity was B. peruviensis, followed by Pseudomonas sp. On the other hand, the treatment that resulted in the least plant development (31% smaller than infected mock) and even increased the severity of the infection was T. longibrachiatum.
番茄褐皱纹病毒(ToBRFV)是影响墨西哥番茄作物的主要病原体之一。尽管努力防止其传播,但由于其通过种子传播的比例较低,且极易通过文化习俗传播,因此几乎不可能做到。因此,人们正在寻求其他管理策略。本研究旨在确定在土壤中施用内生微生物对感染 ToBRFV 的番茄植株的影响。实验以一株番茄为单位,每种处理重复 13 次。对感染 ToBRFV 的番茄植株的处理为 Beauveria peruviencis、Trichoderma longibrachiatum、Pseudomonas sp.和作为病证的水;还包括一个用水处理健康植株的处理作为绝对对照。响应变量为植株高度、气生部分和根部鲜重以及严重程度(两次评估)。采用 Tukey-Kramer HSD 检验对每对测量结果进行分析。发现 Beauveria peruviencis、Trichoderma longibrachiatum、Pseudomonas sp.最有利于受感染植株生长(比受感染的模拟植株高 79%、重 15%)并减轻感染严重程度的处理是 B. peruviensis,其次是假单胞菌 sp.;另一方面,导致植株生长最慢(比受感染的模拟植株小 31%)甚至加重感染严重程度的处理是 T. longibrachiatum。
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引用次数: 0
Weeds and ruderal plants as potential sources of inoculum for vegetable diseases in northern Sinaloa 杂草和灌木植物是锡那罗亚州北部蔬菜病害的潜在接种源
Pub Date : 2023-12-26 DOI: 10.18781/r.mex.fit.2023-4
R. Félix-Gastélum, G. Herrera-Rodríguez, K. Y. Leyva-Madrigal, G. A. Mora-Romero
Weeds and ruderal plants of the families Cucurbitaceae and Solanaceae are addressed as potential sources of inoculum for the development of viral diseases such as Tomato apex necrosis virus (ToANV), zucchini (Zucchini yellow mosaic virus (ZYMV), Watermelon mosaic virus (WMV), Papaya ring spot virus (PRSV-W) and Cucumber mosaic virus (CMV). Reference is made to weeds and ruderal plants as potential sources of inoculum, including wild sunflower for powdery mildew (Golovinomyces spadiceus), wild tobacco for foliar blight (Alternaria spp.), black nightshade for leaf spot (Curvularia moehlemvekiae), Johnson grass for foliar blight (Alternaria sp.), and wild castor bean for foliar blight (Alternaria ricini) and wild melon for downy mildew (Pseudoperonospora cubensis). Future lines of multidisciplinary research focusing on the determination of pathogenicity in cultivated plants of viruses and fungi associated with wild plants and vice versa are proposed; the spatial-temporal distribution of wild plants that may serve as sources of inoculum, as well as the of potential insect vectors of viral diseases, should also be studied. The implementation of modern molecular techniques, such as High Throughput Sequencing, for the detection of phytopathogens is important. All this will contribute to the implementation of environmentally friendly strategies for disease control in agricultural crops in Sinaloa, for the benefit of the vegetable growers.
本文论述了葫芦科和茄科杂草和灌木植物作为番茄顶端坏死病毒 (ToANV)、西葫芦黄镶嵌病毒 (ZYMV)、西瓜镶嵌病毒 (WMV)、木瓜环斑病毒 (PRSV-W) 和黄瓜镶嵌病毒 (CMV) 等病毒病发生的潜在接种源。还提到杂草和灌木植物是潜在的接种源,包括野生向日葵白粉病(Golovinomyces spadiceus)、野生烟草叶枯病(Alternaria spp.叶斑病(Curvularia moehlemvekiae)、叶枯病(Alternaria sp.)的约翰逊草、叶枯病(Alternaria ricini)的野生蓖麻和霜霉病(Pseudoperonospora cubensis)的野生甜瓜。)建议今后开展多学科研究,重点确定与野生植物相关的病毒和真菌在栽培植物中的致病性,反之亦然;还应研究可能作为接种源的野生植物的时空分布以及病毒病的潜在昆虫载体。采用高通量测序等现代分子技术检测植物病原体也很重要。所有这些都将有助于在锡那罗亚州农作物中实施环境友好型病害控制战略,从而造福蔬菜种植者。
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引用次数: 0
Etiology of rhizome rot of asparagus (Asparagus officinalis) in Atenco, Mexico State 墨西哥州阿坦科芦笋(Asparagus officinalis)根茎腐烂病的病原学研究
Pub Date : 2023-11-16 DOI: 10.18781/r.mex.fit.2211-1
The objective of this research was to identify the causal agent of asparagus rhizome rot, as well as evaluate different inoculation methods and the severity of the isolates. Sampling was carried out in five producing plots Atenco, Edo. from Mexico. Five isolates of Fusarium spp. were selected. (one per plot) to perform pathogenicity tests. Three isolates were selected for their colonization characteristics for severity tests with different inoculation methods: Immersion for 12 h, immersion for 30 min and inoculation by contact with absorbent paper soaked in 1 mL of inoculum. Concentrations of 1x106 conidia mL-1 were used. 10 rhizomes were used per treatment and 10 rhizomes without inoculation. To determine the severity, photographs (in GIMP®) of the rhizome were analyzed seven days after inoculation. The isolates were molecularly identified with ITS4/ITS5, EF688/EF1521 and TUBT1/BT2B. Fusarium prolifetatum was morphologically and molecularly identified in the three isolates. The P3DR isolate was the most severe (14.6%), followed by P5DR (13.9%) and P1SIR (11.6%). The most effective inoculation method was immersion for 30 min. They were registered in the NCBI Gene Bank with accessions ON738484 (P3DR), ON973801 (P5DR) and ON738483 (P1SIR). This is the first report of F. prolifetatum in the Edo. from Mexico.
这项研究的目的是确定芦笋根茎腐烂病的病原菌,并评估不同的接种方法和分离物的严重程度。在墨西哥埃多省阿坦科的五个生产地块进行了采样。选择了五个镰刀菌属分离株(每个地块一个)进行病原体鉴定。(每个地块一个)进行致病性测试。根据菌落特征选择了三个分离株,用不同的接种方法进行严重性试验:浸泡 12 小时、浸泡 30 分钟和用浸过 1 毫升接种物的吸水纸接触接种。接种浓度为 1x106 分生孢子 mL-1。每个处理使用 10 个根茎,10 个根茎不接种。为确定严重程度,在接种七天后对根茎的照片(GIMP®)进行分析。分离物通过 ITS4/ITS5、EF688/EF1521 和 TUBT1/BT2B 进行分子鉴定。从形态学和分子鉴定结果来看,这三个分离株中都含有脱落镰刀菌。P3DR 分离物最严重(14.6%),其次是 P5DR(13.9%)和 P1SIR(11.6%)。最有效的接种方法是浸泡 30 分钟。它们在 NCBI 基因库中的登录号分别为 ON738484(P3DR)、ON973801(P5DR)和 ON738483(P1SIR)。这是墨西哥首次报道江户地区的 F. prolifetatum。
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引用次数: 0
Aspergillus oryzae: An opportunity for agriculture 米曲霉:农业的机遇
Pub Date : 2023-11-15 DOI: 10.18781/r.mex.fit.2302-2
Aspergillus oryzae is a filamentous fungus capable of degrading various substances employing enzymes, which is why it is widely used in the biotechnological industry, pharmaceutical products, enzymes for industrial use, bleaching agents, anti-pollution textile treatments. However, few works focus on these microorganism’s field applications. This manuscript reviews the potentially beneficial applications of A. oryzae and some by-products in agriculture as biological control, growth inducer, and bioremediation for soils contaminated with heavy metals.
黑曲霉(Aspergillus oryzae)是一种丝状真菌,能够利用酶降解各种物质,因此被广泛应用于生物技术工业、医药产品、工业用酶、漂白剂、抗污染纺织品处理等领域。然而,很少有著作关注这些微生物的实地应用。本手稿回顾了 A. oryzae 及其一些副产品在农业中作为生物控制、生长诱导剂和重金属污染土壤的生物修复的潜在有益应用。
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Revista Mexicana de Fitopatología, Mexican Journal of Phytopathology
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