First Report of Puccinia menthae Causing Leaf Rust on Spearmint (Mentha spicata) in Mexico.

Spearmint (Lamiaceae) is an aromatic herb widely cultivated in Mexico for its culinary, medicinal, and industrial properties. In May 2020, symptoms and signs of rust were observed in a two-ha commercial crop of spearmint in Cuautla, Morelos (18°50'26.6"N 98°57'31.9"W), Mexico. The disease incidence was 85% and the severity was 23%. Initial symptoms included chlorotic spots on the adaxial surface of the leaves. At advanced stages of the disease, necrotic spots surrounded by chlorotic halos were developed, and later the plants were defoliated. The signs were observed as numerous orange to reddish-brown erumpent pustules primarily on the abaxial surface of the leaves. Microscopic examination of the samples revealed the presence of subepidermal uredinia, erumpent, with hyaline and cylindrical paraphyses. Urediniospores (n = 50) were hyaline to light yellow, globose to obovoid, measuring 17-27  11-25 µm, including 0.6-0.7 µm wall thickness, individually supported on pedicels, echinulate, with two germinative pores. Morphological features of the fungus correspond with previous descriptions of Puccinia mentha by Kabaktepe et al. (2017). A voucher specimen (accession no. UACH448) was deposited in the Department of Agricultural Parasitology Herbarium at the Chapingo Autonomous University. To confirm identification, the 28S gene region of rDNA was amplified from one sample by a nested PCR using the primer sets Rust2inv (Aime, 2006) and LR6 (Vilgalys and Hester, 1990), and Rust28SF primers (Aime et al., 2018), and LR5 (Vilgalys and Hester, 1990) for the first and second reactions, respectively. The sequence of our specimen (GenBank accession No. OL878354) showed 100% homology (923/1304bp) with the type-specimen sequence of P. menthae (GenBank accession No. DQ354513) from Cunila origanoides from USA (Aime, 2006). Also, a phylogenetic analysis (Bayesian inference) including a published 28S dataset for Puccinia species was performed and the isolate UACH448 was grouped into a clade with P. menthae. Pathogenicity was demonstrated by spraying five ml of a suspension of urediniospores (1104 spores/ml) recovered from infected leaves onto leaves of ten healthy spearmint plants and ten noninoculated plants served as controls. All plants were maintained at temperatures from 28 to 35°C and relative humidity ranging from 70 to 80%. All inoculated plants developed the characteristic symptoms of the disease after 14 days, whereas the control plants remained symptomless. The pathogenicity test was performed twice with similar disease response. The morphological characteristics of the pathogen recovered from all the inoculated plants were identical to that originally inoculated, thus fulfilling Koch's postulates. To our knowledge, this is the first report of Puccinia menthae causing leaf rust on Mentha spicata in Mexico. This species has been previously identified in Australia (Edwards et al. 1999), New Zealand (Beresford et al. 1982), and USA (Farr and Rossman, 2021) on Mentha spicata. This disease reduces considerably the quality of peppermint plants, so it is necessary to develop management strategies.

Sorghum Sheath Blight Caused by Fusarium spp. in Sinaloa, Mexico.

Sorghum (Sorghum bicolor) leaf sheath blight was observed for the first time in Sinaloa, Mexico in the summer of 2020. Fungal isolates were obtained from symptomatic tissue in potato dextrose agar. Fusarium spp. were associated with symptomatic plants in 10 sampling sites under field conditions. No root and stalk rot was observed during the sampling period. Analysis of fragments of the translation elongation factor alpha and RNA polymerase II second largest subunit genes indicated that all isolates belong to the Fusarium fujikuroi species complex (FFSC). Five groups were delineated from this complex: F. thapsinum, F. verticillioides, Fusarium sp. (four isolates), Fusarium sp. 4 (Fus4), and Fusarium sp. (Fus16), which is closely related to Fusarium madaense. The morphological characteristics (colony color and morphometry of conidia) of isolates with sequence similarities to those of F. thapsinum and F. verticillioides were in the expected range for these species. The morphology of isolates Fus7a, Fus7b, Fus11, and Fus17, as well as Fus4 and Fus16, were similar to those of the FFSC, specially to F. andiyazi and F. madaense. Inoculations of sorghum with representative isolates of F. thapsinum, F. verticillioides and the unidentified Fusarium species resulted in reddish brown lesions similar to those observed under field conditions; the original isolates inoculated were reisolated fulfilling the Koch's postulates. Although leaf sheaths on sorghum plants were heavily damaged, root and stalk rot were not observed in the greenhouse inoculations or under field conditions. Future research should focus on determining the identity of the unknown Fusarium spp. to design control measures for the disease. This is the first report of Fusarium spp. causing sorghum leaf sheath blight in Mexico.

Rhizospheric bacteria as potential biocontrol agents against Fusarium wilt and crown and root rot diseases in tomato.

The discovery of novel biocontrol agents requires the continuous scrutiny of native microorganisms to ensure that they will be useful on a regional scale. The goal of the present work was to discover novel antagonistic bacteria against Fusarium oxysporum ff. spp. lycopersici race 3 (Fol R3) and radicis-lycopersici (Forl) causing Fusarium wilt disease and Fusarium crown and root rot of tomatoes, respectively. High-throughput liquid antagonism screening of 1,875 rhizospheric bacterial strains followed by dual confrontation assays in 96-well plates was used to select bacteria exhibiting > 50% fungal growth inhibition. In a second dual confrontation assay in 10-cm Petri dishes, bacteria showing > 20% Fol R3 or Forl growth inhibition were further screened using a blood hemolysis test. After discarding ß-hemolytic bacteria, a seedling antagonistic assay was performed to select five potential antagonists. A phylogenetic analysis of 16S rRNA identified one strain as Acinetobacter calcoaceticus (AcDB3) and four strains as members of the genus Bacillus (B. amyloliquefaciens BaMA26, Bacillus siamensis BsiDA2, B. subtilis BsTA16 and B. thuringiensis BtMB9). Greenhouse assays demonstrated that BsTA16 and AcDB3 were the most promising antagonists against Fol R3 and Forl, respectively. Pathogen biocontrol and growth promotion mechanisms used by these bacteria include the production of siderophores, biofilm, proteases, endoglucanases and indole acetic acid, and phosphate solubilization. These five bacteria exerted differential responses on pathogen control depending on the tomato hybrid, and on the growth stage of tomatoes. We report for the first time the use of an Acinetobacter calcoaceticus isolate (AcDB3) to control Forl in tomato under greenhouse conditions.

First Report of Golovinomyces sonchicola Causing Powdery Mildew on Sonchus oleraceus in Mexico.

Sonchus oleraceus, common sow thistle, is native to Europe, Northern Africa, and Western Asia. This plant has become a common weed throughout the world. In Mexico, this weed has become widely naturalized by replacing indigenous plants and invading many agricultural areas. During the spring of 2018 and 2019, common sow thistle plants showing typical symptoms and signs of powdery mildew, were collected from agricultural fields in Ahome, Sinaloa, Mexico. As much as 30% of plants were diseased and 60 to 95% of the foliage was affected. Mycelium was conspicuous and white-gray, and on stems and both surfaces of leaves. Appressoria were nipple-shaped to crenulate. Conidiophores (n= 30) were hyaline, cylindrical, erect, and up to 150 µm long. Foot-cells (n= 30) were distinctly curved, 47 to 75 × 10 to 13 µm, slightly constricted, followed by 1-3 shorter cells and formed conidia in chains. Conidia (n= 100) were ellipsoid to doliiform to subcylindrical, 28 to 37 × 14 to 19 µm, lacked fibrosin bodies, and germinated from the apex. Chasmothecia were not observed. The morphological characters were consistent with those of the anamorphic state of Golovinomyces sonchicola (Braun and Cook 2012, Jakse et al. 2019). A voucher specimen (accession no. FAVF215) was deposited in the Herbarium of the Faculty of Agriculture of El Fuerte Valley at the Autonomous University of Sinaloa (Juan Jose Rios, Sinaloa, Mexico). To confirm the morphological identification, genomic DNA was extracted from mycelium and conidia, and the internal transcribed spacer (ITS) region and part of the 28S gene were amplified by PCR and sequenced. The ITS region of rDNA was amplified using the primers ITS5/ITS4 (White et al. 1990). For amplification of the 28S rRNA partial gene, a nested PCR was performed using the primer sets PM3 (Takamatsu and Kano 2001)/TW14 (Mori et al. 2000) and NL1/TW14 (Mori et al. 2000) for the first and second reactions, respectively. Phylogenetic analyses using the maximum parsimony and maximum likelihood methods (Braun et al. 2019), including ITS and 28S sequences of isolates of Golovinomyces spp. were performed and confirmed the results obtained from the morphological analysis. Isolate FAVF215 grouped in a clade with the other isolates of G. sonchicola. The ITS and 28S sequences were deposited in GenBank under accession numbers MW425872 and MW442972, respectively. Pathogenicity was demonstrated by gently dusting conidia from infected leaves onto leaves of 20 healthy plants and covered with plastic bags for 24 h. Ten non-inoculated plants served as controls. All plants were maintained in a greenhouse at 25 to 35ºC. All inoculated plants developed similar symptoms to those observed in the field from natural infections after 12 days, whereas powdery mildew symptoms and signs were not observed on control plants. The morphology asexual structures of fungus on inoculated plants were identical to those on naturally infected plants, fulfilling Koch's postulates. Inoculation tests were repeated twice with identical results. Based on the morphological data and phylogenetic analysis, the fungus was identified as G. sonchicola. This fungus has been reported causing powdery mildew on S. oleraceus in Germany, The Netherlands, Slovenia, and The United Kingdom (Farr and Rossman 2021). To the best of our knowledge, this is the first report of G. sonchicola causing powdery mildew on S. oleraceus in Mexico. This powdery mildew pathogen may represent an option for the biological control of common sow thistle.

First Report of Powdery Mildew on Leucophyllum frutescens Caused by Podosphaera xanthii in Mexico.

Leucophyllum frutescens (Scrophulariaceae family), commonly known as Texas sage or cenizo, is an evergreen shrub native to southwestern United States and northern Mexico. This plant is commercially sold as a native, drought-tolerant ornamental. During the spring of 2019 and 2020, typical symptoms of powdery mildew were found on cenizo plants growing as ornamentals in urban areas in the municipality of Ahome, Sinaloa, Mexico. Disease incidence was 95% from a sampled population of 120 plants. Initial symptoms of powdery mildew developed as irregular white colonies on upper leaf surfaces which expanded as infections progressed. In severe infections, leaves became distorted, exhibiting premature defoliation. Microscopic examination showed nipple-shaped appressoria. Conidiophores (n= 30) were hyaline, cylindrical, erect, 89.4 to 134.2 µm long, and forming catenescent conidia. Foot-cells were cylindrical, 35.7 to 65.3 × 10.2 to 13.5 µm, followed by 1-3 shorter cells. Conidia (n= 100) were hyaline, ellipsoid to ovoid, 27.9 to 40.5 × 13.8 to 18.9 µm, containing distinct fibrosin bodies. Germ tubes were simple to forked and laterally produced from the middle of conidia. Chasmothecia were not found during the sampling period on the infected leaves. Based on morphological characteristics, the fungus was identified as Podosphaera xanthii (Braun and Cook 2012). A voucher specimen (accession no. FAVF219) was deposited in the Herbarium of the Faculty of Agronomy of El Fuerte Valley at the Autonomous University of Sinaloa (Juan Jose Rios, Sinaloa, Mexico). To further confirm the identification, total DNA was extracted, and the internal transcribed spacer (ITS) region was amplified by PCR using the primers ITS5/ITS4 (White et al. 1990) and sequenced. The resulting 503 bp sequence (GenBank accession no. MT624793) had 100% coverage and 100% identity to those of P. xanthii (MT568609-MT568611, MT472035, MT309699, MT250855, MT242593). A phylogenetic tree using the maximum parsimony (MP) and maximum likelihood (ML) methods and including published ITS sequences for Podosphaera species was obtained. Phylogenetic analyses revealed that ITS sequence from FAVF219 isolate was grouped into a clade with P. xanthii. Pathogenicity was demonstrated by gently dusting conidia from infected leaves onto 50 leaves of five healthy plants. Five non-inoculated plants served as controls. All plants were covered with polyethylene bags for 48 h to maintain high humidity and were maintained in a greenhouse at temperatures ranging from 20 to 35ºC. All inoculated plants developed similar symptoms to the original observations after 19 days, whereas no symptoms of powdery mildew were observed on control plants. The fungus present on the inoculated plants was morphologically identical to that originally observed on diseased plants, fulfilling Koch's postulates. This fungus has been reported infecting members of the Cucurbitaceae in Mexico (Félix-Gastélum et al. 2017; Farr and Rossman 2020). However, to our knowledge, this is the first report of P. xanthii causing powdery mildew on a member of Scrophulariaceae, specifically L. frutescens in Mexico and worldwide. Further studies for monitoring and control strategies of powdery mildew on Texas sage are required.

Draft Genome Sequence of a Streptomycete Isolated from Potato Common Scab Lesions in the State of Sinaloa, Mexico.

Streptomyces sp. strain V2 was isolated from potato scab lesions in the state of Sinaloa, Mexico, and appears to be responsible for outbreaks in the area. The thaxtomin cluster was found in the ∼10.2-Mb genome; this cluster is associated with potato common scab disease in other potato pathogens.

Rhizospheric bacteria of maize with potential for biocontrol of Fusarium verticillioides.

The stalk, ear and root rot (SERR) of maize caused by Fusarium verticillioides (Fv) severely impacts crop production in tropical and subtropical regions. The aim of the present work was to screen bacterial isolates in order to find novel native biocontrol agents against Fv. A culturable bacterial collection consisting of 11,520 isolates enriched in Firmicutes and Proteobacteria was created from rhizosphere samples taken from SERR symptomatic or asymptomatic maize plants. The complete collection was screened for potential activity against Fv using a liquid antagonism assay followed by dual cultures in solid medium, selecting for 42 bacteria (Bacillus, Pseudomonas and Paenibacillus) that inhibit Fv growth (>45 %). In planta assays demonstrated that three Bacillus isolates: B. megaterium (B5), B. cereus sensu lato (B25) and Bacillus sp. (B35) displayed the highest antagonistic activity against Fv. Pot experiments performed in a greenhouse with Bacillus cereus sensu lato B25 confirmed these findings and showed a reduction of Fv disease severity and incidence on plants. Antagonistic activity analysis revealed that these strains produce glucanases, proteases or chitinases, as well as siderophores and auxins and suggests these as possible control mechanisms against Fv.

Cytogenetic biomonitoring of occupationally exposed workers to ashes from burning of sugar cane in Ahome, Sinaloa, México.

Burning the sugar cane field before harvesting has a negative impact on both air and human health, however this issue had not been explored in Mexico. The objective of this work was to determine the chromosomal damage in workers from sugar cane burning fields in Sinaloa, México. To this purpose, we analyzed 1000 cells of buccal exfoliated epithelia from 60 exposed workers and 60 non-exposed controls to determine micronucleus frequencies and other nuclear abnormalities. The results indicated significant higher values of micronucleus and nuclear abnormalities such as binucleate cells, pyknosis, karyolysis, chromatin condensation and nuclear buds frequencies in the exposed subjects compared to those that were not exposed. Our data indicates that sugar cane burning, that generates polycyclic aromatic hydrocarbons, represents a genotoxic risk for workers in this important sugar cane producing area in Mexico.

Genotoxic biomonitoring of agricultural workers exposed to pesticides in the north of Sinaloa State, Mexico.

Genotoxic damage was evaluated in 70 agricultural workers, 25 women and 45 men, exposed to pesticides in Las Grullas, Ahome, Sinaloa, Mexico, with an average of 7 years of exposure. The effect was detected through the sister chromatid exchanges (SCE) in lymphocytes of peripheral blood and micronuclei (MN) and other nuclear anomalies (NA) in buccal exfoliated cells. Also, the influence on cellular proliferation kinetics (CPK) was studied by means of the replication index (RI) and the cytotoxic effect was examined with the mitotic index (MI). The non-exposed group consisted of 70 other persons, 21 women and 47 men from the city of Los Mochis, Sinaloa, Mexico. Significant differences between the exposed and the non-exposed groups were observed in SCE, CPK, MI, MN and NA. Analysis of variance revealed that age, gender, smoking and alcohol consumption did not have a significant effect on genetic damage. However, there was a correlation between exposure time to pesticides and SCE frequency. These results could have been due to the exposure of workers to pesticides containing different chemical compounds. This study afforded valuable data to estimate the possible risk to health associated with pesticide exposure.