A multiplex reverse transcription polymerase chain reaction (RT-PCR) assay was developed to simultaneously detect bean common mosaic virus (BCMV), bean common mosaic necrotic virus (BCMNV), and bean golden yellow mosaic virus (BGYMV) from common bean leaves dried with silica gel using a single total nucleic acid extraction cetyl trimethyl ammonium bromide (CTAB) method. A mixture of five specific primers was used to amplify three distinct fragments corresponding to 272 bp from the AC1 gene of BGYMV as well as 469 bp and 746 bp from the CP gene of BCMV and BCMNV, respectively. The three viruses were detected in a single plant or in a bulk of five plants. The multiplex RT-PCR was successfully applied to detect these three viruses from 187 field samples collected from 23 municipalities from the states of Guanajuato, Nayarit and Jalisco, Mexico. Rates of single infections were 14/187 (7.5%), 41/187 (21.9%), and 35/187 (18.7%), for BGYMV, BCMV, and BCMNV, respectively; 29/187 (15.5%) samples were co-infected with two of these viruses and 10/187 (5.3%) with the three viruses. This multiplex RT-PCR assay is a simple, rapid, sensitive, and cost-effective method for detecting these viruses in the common bean and can be used for routine molecular diagnosis and epidemiological studies.
Viruses are an important disease source for beans. In order to evaluate the impact of virus disease on Phaseolus biodiversity, we determined the identity and distribution of viruses infecting wild and domesticated Phaseolus spp. in the Mesoamerican Center of Domestication (MCD) and the western state of Nayarit, Mexico. We used small RNA sequencing and assembly to identify complete or near-complete sequences of forty-seven genomes belonging to nine viral species of five genera, as well as partial sequences of two putative new endornaviruses and five badnavirus- and pararetrovirus-like sequences. The prevalence of viruses in domesticated beans was significantly higher than in wild beans (97% vs. 19%; p < 0.001), and all samples from domesticated beans were positive for at least one virus species. In contrast, no viruses were detected in 80–83% of the samples from wild beans. The Bean common mosaic virus and Bean common mosaic necrosis virus were the most prevalent viruses in wild and domesticated beans. Nevertheless, Cowpea mild mottle virus, transmitted by the whitefly Bemisia tabaci, has the potential to emerge as an important pathogen because it is both seed-borne and a non-persistently transmitted virus. Our results provide insights into the distribution of viruses in cultivated and wild Phaseolus spp. and will be useful for the identification of emerging viruses and the development of strategies for bean viral disease management in a center of diversity.
Viral diseases pose a significant threat to tomato crops (Solanum lycopersicum L.), one of the world’s most economically important vegetable crops. The limited genetic diversity of cultivated tomatoes contributes to their high susceptibility to viral infections. To address this challenge, tomato breeding programs must harness the genetic resources found in native populations and wild relatives. Breeding efforts may aim to develop broad-spectrum resistance against the virome. To identify the viruses naturally infecting 19 advanced lines, derived from native tomatoes, high-throughput sequencing (HTS) of small RNAs and confirmation with PCR and RT-PCR were used. Single and mixed infections with tomato mosaic virus (ToMV), tomato golden mosaic virus (ToGMoV), and pepper huasteco yellow vein virus (PHYVV) were detected. The complete consensus genomes of three variants of Mexican ToMV isolates were reconstructed, potentially forming a new ToMV clade with a distinct 3’ UTR. The absence of reported mutations associated with resistance-breaking to ToMV suggests that the Tm-1, Tm-2, and Tm-22 genes could theoretically be used to confer resistance. However, the high mutation rates and a 63 nucleotide insertion in the 3’ UTR, as well as amino acid mutations in the ORFs encoding 126 KDa, 183 KDa, and MP of Mexican ToMV isolates, suggest that it is necessary to evaluate the capacity of these variants to overcome Tm-1, Tm-2, and Tm-22 resistance genes. This evaluation, along with the characterization of advanced lines using molecular markers linked to these resistant genes, will be addressed in future studies as part of the breeding strategy. This study emphasizes the importance of using HTS for accurate identification and characterization of plant viruses that naturally infect tomato germplasm based on the consensus genome sequences. This study provides crucial insights to select appropriate disease management strategies and resistance genes and guide breeding efforts toward the development of virus-resistant tomato varieties.
Tomato (Solanum lycopersicum L.) is one of the most important cultivated vegetables in the world. However, in some countries such as Mexico the lack of cultivars adapted to different environmental production conditions is a limitation. Moreover, recent studies have indicated that breeding aimed at increasing yield has led to a loss of genetic diversity. Therefore, it is necessary to explore and characterize new sources of germplasms. This study aimed to characterize new sources of germplasm and identify the most transcendental traits for distinguishing tomato types and lines that are useful for the genetic improvement of the species. Sixty characters were evaluated in 16 advanced lines of native tomatoes from Central and Southern Mexico during the fall–winter cycles 2023–2024 at the Bajío Experimental Station, Celaya, Guanajuato, Mexico, based on the guidelines of the International Union for the Protection of New Varieties of Plants (UPOV) and the International Plant Genetic Resources Institute (IPGRI). The data were analyzed using descriptive statistics, analysis of variance and post hoc tests, canonical discriminant analysis, and the Eigenanalysis selection index method (ESIM). Morphological variation showed that five qualitative traits were determinant factors in distinguishing tomato types and lines, whereas agronomic discriminant traits were the equatorial and polar diameters of the fruit and its ratio, number of locules, pedicel length, stem length, and internode distance. In addition, significant positive correlations were found between leaf length and width, equatorial diameter of the fruit, and polar diameter of the fruit. Lines JCM-17, JMC-10, and JCM-01 were the most selectable lines according to the ESIM values. The morphological variation found and the characteristics with higher selection values identified may be valuable for optimizing the tomato genetic improvement process in general.
In the context of a regional project with the aim of identifying viruses infecting cultivated Phaseolus vulgaris (CB) and wild bean (WB): P. vulgaris, P. coccineus and P. leptostachyus species in the central-western region of Mexico, six leaves from 394 CB and 245 WB plants with (74%) and without (26%) viral symptoms were collected in the Mexican states of Guanajuato, Jalisco and Nayarit between 2013-2015. The symptoms observed were chlorotic lesions, mild and severe mottling, mild mosaics and leaf deformation. To identify the viruses, small RNA sequencing and an assembly technique was used (Kreuze, 2014). Small RNA (sRNA) reads with identity to Cowpea mild mottle virus (CPMMV) and Bean common mosaic necrosis virus (BCMNV) were identified in the sRNA library CN2Ls prepared from one sample of Phaseolus vulgaris cv. ‘Coranay’ collected in Nayarit state (CN2, NCBI BioProject: PRJNA362733), whose leaves exhibited mottling, mild mosaic and slight deformation symptoms. After bioinformatic processing and assembl...