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In the present study, a droplet digital PCR assay was developed for detection of Tomato brown rugose fruit virus, a new Tobamovirus of tomato and other solanaceous plants, which expands the diagnostic strategies for this pathogen. Candidate reference DNA material was also obtained to be employed as positive control in tomato and pepper samples. Recombinant plasmids encode for ToBRFV coat protein (CP-ToBRFV) gene and Solanum lycopersicum GAPDH fragments, and CP-ToBRFV and Capsicum annuum GAPDH. To our knowledge, this is the first report of ToBRFV detection in tomato and pepper seeds using ddPCR., (Copyright © 2022 Elsevier B.V. All rights reserved.)

Infectivity of an alfalfa mosaic virus (AMV) isolate from Leonotis nepetaefolia in different tomato cultivars was analyzed. Symptoms typical of AMV infection were observed in indicator plants, but not in Flora Dade and Rio Grande tomato cultivars; however, mild symptoms were observed in cv. Rutgers. Furthermore, at least 1 kb of the 3´ segment of RNA 2 and the coat protein gene were missing in systemic leaves of inoculated Rio Grande and Flora Dade plants, while in cv. Rutgers infected with this AMV strain all genomic components were detected. Northern blot analysis of plants infected with the aforementioned AMV isolate confirmed the absence of the CP gene, but suggested rearrangements in both RNA 2 and 3. Factors that may affect differential movement or systemic accumulation of genomic components in multipartite viruses in plants are discussed., (Copyright © 2019 Elsevier B.V. All rights reserved.)

The complete genome sequence of a trichovirus was obtained from peach samples collected from Mexico and found to be 7985 nucleotides long, excluding the poly(A) tail. Phylogenetic analysis using the complete nucleotide sequence revealed that the virus is a member of the genus Trichovirus and is closely related to peach mosaic virus (PcMV) and cherry mottle leaf virus (CMLV). The highest nucleotide sequence identity was 70% to both PcMV and CMLV, indicating that this virus, which we have tentatively named "peach virus M" (PeVM) should be considered a member of a new trichovirus species. We determined, for the first time, the initiation sites of the subgenomic RNAs (sgRNA) of a trichovirus. The sgRNAs for the movement and coat proteins started with the sequence 'GAA', while the smallest one, coding for the nucleotide-binding protein, started with the nucleotides 'GU'. In all cases, the sgRNAs leader ranged between 113 and 121 nt in length.

Neobuxbaumia tetetzo (Coulter) Backeberg (tetecho) is a columnar cactus endemic to Mexico. Tetecho plants, flowers, fruits, and seeds play an important role in the semiarid ecosystem, as they serve as a refuge and food for insects, bats, and birds, and are widely used by ethnic groups since pre-Hispanic times. Tetecho is affected by a soft rot that damages the whole plant and causes its fall and disintegration. Eight bacterial colonies of similar morphology were isolated from plants showing soft rot and inoculated in healthy tetecho plants, reproducing typical symptoms of soft rot 9 days after inoculation. Ten representative isolates were selected for phenotypic and genetic identification using 16s rDNA, IGS 16S-23S rDNA, and rpoS genes and for pathogenicity tests on several members of the cactus family and other plants. Based on the results, these bacterial isolates were identified as Pectobacterium carotovorum subsp. brasiliense. Inoculation of this bacteria caused soft rot in different cacti, fruits, leaves, and roots of other plants. This is the first report of the subspecies brasiliense of P. carotovorum causing soft rot and death in cacti in the world and the first report of this subspecies in Mexico.

During 2001, branch dieback, black trunk rot, and resinosis were observed on mesquite in the biosphere reserve of Tehuacan, Mexico (18°15'N, 97°25'W) A light brown growth, which included Nodulosporium-like conidiosphores and hyaline conidia that were green in mass and ellipsoid with one end truncate developed on diseased branches. Below the conidiophores and conidia, glomerate to pulvinate stromata formed with conspicuous, black, perithecial mounds with globose perithecia. Ascospores were dark brown, unicellular, ellipsoid, nonequilateral, with narrowly rounded ends, a straight germ slit with a perispore that was dehiscent in 10% KOH, and a conspicuous coil-like, smooth epispore. Sexual reproduction was induced on sterile toothpicks in potato dextrose agar, malt extract agar, or V8 agar (with 10% calcium chloride). The fungus was identified as Hypoxylon diatrypeoides Rehm (1). Samples of mesquite branches with stromata of H. diatrypeoides were deposited in the J. H. Miller Herbarium of the University of Georgia (GAM16048). During the summer of 2002, three pathogenicity tests were performed under greenhouse conditions using three healthy young mesquite plants (25 cm high) per treatment per test. The treatments were: (i) inoculation of branches by wounding with a colonized toothpick from V8 agar, covered with mycelium and perithecia; (ii) spraying ascospores on branches previously wounded with a sterile toothpick; (iii) spraying ascospores on unwounded plants; (iv) plants wounded with sterile toothpicks; and (v) unwounded and uninoculated plants. Fifteen days after inoculation, branch dieback and black trunk rot symptoms were induced in 100% of mesquite plants inoculated with toothpicks and in 50% of wounded plants inoculated with ascospores. No symptoms were seen in the unwounded plants and control treatments. H. diatrypeoides was reisolated from the symptomatic branches. Previously, the fungus had been reported only from the Southern Hemisphere (Brazil and New Zealand), but to our knowledge, this is the first report from Mexico and the Northern Hemisphere. This is also the first evidence of its role as a plant pathogen. Reference: (1) Y. M. Ju and J. D. Rogers. A revision of the genus Hypoxylon. Mycologia Memoir No 20. The American Phytopathological Society, St. Paul, MN, 1996.