First report of Irpex rosettiformis causing white root rot in avocado trees in Michoacán, México.

In México, avocado production is an important economic source. In the last season it generated $ 3. 27 billion USD of foreign currency in the country. Irpex spp. are wood decay fungi. In the period 2019-2022, in the state of Michoacán (19°13' N; 101°55' W), México, basidiomes of Irpex sp. were observed on the base of trunks and crowns of 5-years-old and older avocado (Persea americana) trees. The trees exhibited disease symptoms that included white root rot, leaf yellowing, small leaves, branch diebacks, generalized defoliation, apical flaccidity, abundant but small sun burnt fruits due to the lack of foliage, and after 2-4 years of first disease appearance, the infected trees died. In the place where fungus was established, abundant white and cottony mycelium was formed, which caused trees decay. The incidence of the disease in the sampled orchards was estimated to be 30% per ha with 350 - 400 trees, which was determined through a simple sampling design focused on trees with signs and symptoms of the disease due to the phytopathogen. Samples of infected tissue (roots and stems) and fungal basidiomes were collected from 90 trees (5-6 per orchard). The symptomatic avocado trees studied were randomly selected from 17 orchards. For the fungal macroscopic characterization, the synoptic keys described by Gilbertson and Ryvarden (1986) and by Largent (1973) were used. The samples showed typical structures corresponding to Irpex sp., including rosettes, annual basidiomes, a system of monomitic hyphae, and subglobose basidiospores. In vitro fungal isolation from basidiomes and infected tree tissues was done according to the protocol of Agrios (2004). The fungal strains were maintained on PDA at 28 °C. At 16 days of incubation the colonies were opaque, whitish with fluffy and corky mycelium. Microscopic analysis of the fungus showed typical yellowish spores, with an ellipsoid shape of 3-4 x 4-5.5 µm (50 accounted structures per isolate [N=19]) and basidia of 20-25 x 4.5-5.5 µm (n=20 basidiomes). For molecular characterization, two molecular markers were used, the internal transcribed spacer rDNA-ITS1 5.8 rDNA-ITS2 (ITS; White et al. 1990) and the large ribosomal subunit (LSU; Vilgalys and Hester 1990). The PCR reaction was performed as described by Martínez-González et al. (2017). The consensus sequences were compared with those deposited in the NCBI-GenBank, using the BLASTN 2.2.19 tool (Zhang et al. 2000), the samples showed 99% match with the species, Irpex rosettiformis. GenBank accession numbers of the submitted isolates are summarized in supplementary Table 4. To test Koch's postulates, 3-months old avocado plants grown in greenhouse conditions were inoculated (n = 10 per each isolate [N= 19]) on the roots with 3 g of I. rosettiformis mycelium. The experiment was done twice with 20 non-inoculated plants as control. After 67 days, basidiomes (50 x 70 x 1.5 mm in average) were observed where the disease incidence was >77%, with subsequent tree decline. The pathogen was re-isolated in vitro in PDA and its identity was confirmed by morphological characteristics of mycelium. This work shows that I. rosettiformis is not only a wood decay fungus, but also a phytopathogen, the causative agent of white root rot disease in P. americana var. drymifolia, cultivar 'Hass', which establishes a precedent for monitoring and preventing its proliferation to other regions in the American continent and the world where nursery avocado seedlings are exported.

Fusarium pseudocircinatum causing stunting and malformation of sunflower plants in Brazil.

Sunflower (Helianthus annuus L.) is among the main oleaginous crops used in Brazil. During January, 2017, at CCA/UFPB laboratory and greenhouses (Areia/Brazil, 6°58'12″ S; 35°42'15″ W), we observed various sunflower seeds (cultivar Olisun 3, 2017-2018 crop) highly infested with Fusarium. Those seeds were from crops in the municipality of Alagoinha -PB/Brazil (06º57'00'' S; 35º32'42'' W), supplied by Empresa Brasileira de Pesquisa Agropecuária/EMBRAPA. The emerged seedlings from these seeds were also contaminated, with 5% to 26% of them exhibiting stunting and malformation. Fusarium strains were isolated from symptomatic plants, and a single spore was used to grow pure colonies on potato-dextrose-agar (PDA) and synthetic-nutrient-poor-agar (SNA) media. Mycelia of PDA colonies were floccous and dense varying from yellow to orange. Fungal colonies developed aerial mycelium, producing orange pigments. On SNA, hyaline macroconidia, measuring 2.9-4.1 x 32.4-65.0 µm, slightly falcate with three to six septa. Oval microconidia, measuring 2.4-3.6 x 5.1-9.0 µm, were abundant in false heads forming on monophyalides. Chlamydospores were absent. Sterile hyphae were rarely formed. Colectively, the morphological features corresponded to species that belong to the Fusarium fujikuroi species complex (Leslie & Summerell, 2006). To assure the species identity, we sequenced the elongation factor 1α region of two representative isolates (i.e., F2 and F3, GenBank access numbers: MZ666934 and MZ666935, respectively) and compared them to the other Fusarium species found at Fusarium-ID and GenBank databases. Subsequently, we performed a maximum likelihood phylogenetic analysis including previously published sequences (Nicolli et al., 2020). Both isolates exhibited 100% similarity with Fusarium pseudocircinatum (MN386745), and clustered with its ex-type at 100% bootstrap values. The isolates were then grown on PDA amended with manitol to adjust the osmotic pressure to -1.0 Mpa, at 25 ± 2 ° C, for seven days (Sousa et al., 2008). A total of 100 disinfested sunflower seeds (cultivar Olisun 3, 2018-2019 crop) were distributed over the colonies and 48h later they were sown on sterile substrate maintained inside a greenhouse. About 30 days after inoculation, the emerged plants exhibited symptoms of stunting and malformation (60%) compared to controls, which were healthy. F. pseudocircinatum was reisolated from the symptomatic plants, completing Koch's postulates and identified based on above morphological and molecular biological methods. This test was performed twice. Fusarium pseudocircinatum is a broadly distributed and ecologicaly diverse species that infects several wild and cultivated plants. For instance, it was reported on seeds of the wild 'Peroba Rosa' (Aspidosperma polyneuron Muell. Arg.) in Brazil (Mazarotto et al. 2020). Infection of sunflowers may cause plant stand failures, thus resulting in yield and economic losses for Brazilian growers. The correct identification of any pathogen, especialy a generalist one such as F. pseudocircinatum, is crucial to develop eficient management strategies. To our best knowledge, this is the first report of F. pseudocircinatum causing stunting and malformation of sunflower plants in Brazil.

First Report of Xanthomonas citri subsp. citri causing Citrus Canker on lime in Rio Grande do Norte, Brazil.

Citrus canker caused by Xanthomonas citri subsp. citri is one of the most important citrus diseases in the world (Gottwald et al. 2002), mainly for citrus-producing countries with humid sub-tropical regions such as United States, Argentina, and Brazil, where losses may be significant (Behlau et al. 2020). In the state of Rio Grande do Norte (RN), Brazil, citrus production is expanding and shows social and economic importance for small farmers, which produced approximately 297 tons of lime in this state in 2019 (IBGE 2021). In December 2019, we observed symptoms of erumpent lesions with margins surrounded by yellow haloes on leaves and fruit of the lime (Citrus aurantifolia cv. 'Galego') (about 5% incidence) in a plantation located in the municipality of Mossoró, RN (05°12'21.1"S, 37°19'16"W). Samples were collected from the lime orchard, and five bacterial strains (CCRMXC01 to CCRMXC05) showing yellow, convex, mucoid colonies were isolated in a nutrient-yeast-dextrose-agar medium (NYDA). Pathogenicity tests were performed on sweet orange (C. sinensis cv. 'Pêra') and lime (C. latifolia cv. 'Tahiti') seedlings. Four wounds per leaf (upper side) were carried out with an entomological pin and 10 µl of a bacterial suspension (108 CFU mL-1) were deposited on each wound. The negative control consisted of leaves treated with sterile distilled water (SDW). For each citrus species, we used four replicates per strain and one leaf with four wounds per replicate. Inoculated leaves developed erumpent lesions with margins surrounded by yellow haloes six days after inoculation (DAI) in both citrus species, while leaves treated with SDW remained symptomless. Nine DAI, we reisolated the pathogen and performed rep-PCR (REP, ERIC, and BOX-PCR) analyses (Gama et al. 2018) with the strains inoculated and reisolated to confirm the identity of the strains and to fulfill Koch's postulates. The strains were stored at the Culture Collection Rosa Mariano (CCRM) of the Phytobacteriology Laboratory at the Universidade Federal Rural de Pernambuco. The five strains reisolated showed the same REP, ERIC, and BOX-PCR profiles as the strains used for inoculations. The molecular identification was performed sequencing the dnaK, fyuA, gyrB, and rpoD genes (Young et al. 2008). Each fragment was sequenced in both the forward and reverse directions. Using the BLASTn tool, we observed that sequences of the dnaK (GenBank MW218913 to MW218917), fyuA (GenBank MW218918 to MW218922), and rpoD (GenBank MW218928 to MW218932) genes of the strains CCRMXC01 to CCRMXC05 showed 100% of identity with the sequences of these genes from the type strain (ICMP 24T) and of other strains of X. citri subsp. citri (ICMP 21 and ICMP 7493), while sequences of gryB (GenBank MW218923 to MW218927) of the former strains showed 100% identity with the gyrB sequence of the strains ICMP 24T and ICMP 7493 and 99,85% identity with strain ICMP 21. This short variation in the sequence of the gyrB gene also may be observed among strains of X. citri subsp. citri available in NCBI database (https://www.ncbi.nlm.nih.gov/). The phylogenetic analysis performed using Bayesian inference and the concatenated sequence of all the type or representative strains of species and pathovars of Xanthomonas available in GenBank showed that the strains CCRMXC01 to CCRMXC05 clustered together with strain ICMP 24T with 1.0 posterior probability. To our information, this is the first report of X. citri subsp. citri causing citrus canker on lime in RN state, Brazil.

A Threshold-Based Decision-Support System for Fungicide Applications Provides Cost-Effective Control of Citrus Postbloom Fruit Drop.

Postbloom fruit drop (PFD) of citrus, caused by Colletotrichum acutatum sensu lato and C. gloeosporioides sensu lato, is an important disease in the humid tropics of the American continent. PFD mainly affects flowers, on which typical symptoms are characterized by orange-brown lesions with presence of acervuli. The disease has a sporadic occurrence, but preventative fungicide sprays are applied every season. The objective of this study was to evaluate the effectiveness of a fungicide spray strategy for PFD based on a predictive model of C. acutatum conidium germination linked to weather conditions. Fungicide sprays were performed when the model predicted pre-established thresholds of 10, 15, 20, and 25% of germinated spores (T10, T15, T20, and T25, respectively). Five experiments were conducted in two different seasons in the state of São Paulo, Brazil. PFD control efficacy of the threshold-based treatments was compared with a nontreated control and to a calendar-based spray system. Additionally, an economic analysis was performed to assess the gross income revenues of the fungicide spraying strategies. Disease control in plots treated at T10, T15, and T20 was as effective as the calendar-based strategy. The number of fungicide applications was reduced by 33 to 71% when sprays were applied at T15 and T20, and gross income increased or was comparable to that of the other treatments. Therefore, using a conidium germination model with a threshold of 15 or 20% is recommended as a spraying strategy for PFD management in Brazil.

Agrometeorology and plant disease management: a happy marriage

Many plant disease outbreaks are triggered by suitably warm temperatures during periods of leaf wetness. Measurements or estimations of leaf wetness duration provided by Agrometeorologists have allowed Plant Pathologists to devise weather timed spray schemes which often reduce the number of sprays required to control plant diseases, thus lowering costs and benefitting the environment. In the near future, tools such as numerical weather models with small grid spacings, and improved weather radar, are expected to reduce the need for tight networks of surface observations. The weather models will also provide growers with forecast warnings of potential upcoming disease outbreaks, which will further enhance the contribution of agrometeorology to plant disease management.

A disseminação de muitas doenças de plantas é influenciada por condições favoráveis de temperatura durante o período de molhamento foliar. As medidas e estimativas da duração do período de molhamento foliar fornecidas pelos Agrometeorologistas têm permitido aos Fitopatologistas dar alertas sobre a necessidade de pulverizações com base nas condições meteorológicas, o que normalmente reduz o número de aplicações para o controle de doenças, resultando em menor custo de produção e menor contaminação do ambiente. Em um futuro próximo, ferramentas como os modelos numéricos de tempo, com alta resolução espacial, e os radares meteorológicos mais avançados, deverão reduzir a necessidade de redes de observação meteorológica de superfície mais densas. Os modelos meteorológicos também possibilitarão a previsão de disseminação potencial das doenças de plantas, o que irá aumentar ainda mais a contribuição da agrometeorologia para o controle fitossanitário mais racional.

Performance of cylindrical leaf wetness duration sensors in a tropical climate condition

Leaf wetness duration (LWD) measurements are required for disease warning in several agricultural systems, since it is an important variable for the diagnose of plant disease epidemiology. The cylindrical sensor is an inexpensive and simple electronic LWD sensor initially designed to measure this variable for onions, however some studies show that it may be helpful for standard measurements in weather stations and also for different crops. Therefore, the objective of this study was to assess their performance under tropical climate conditions, in Brazil, having as standard measurements those obtained by flat plate sensors, which have presented very good performance when compared with visual observations. Before field assessments, all LWD sensors used in our study (flat plates and cylinders) were white latex painted and submitted to a heat treatment. Laboratory tests were performed in order to determine the resistance threshold for the sensor to be considered wet and the time response of the sensors to wetness. In the field, all cylindrical sensors were initially deployed horizontally 30-cm above a turfgrass surface in order to assess the variability among them with respect to LWD measurements. The variability among the horizontal cylindrical sensors was reduced by using a specific resistance threshold for each sensor. The mean coefficient of variation (CV) of LWD data measured by the cylindrical sensors was 9.7%. After that, the cylindrical sensors were deployed at five different angles: 0º, 15º, 30º, 45º, and 60º. Data of measurements made at these angles were compared with the standard measurement, obtained by flat plate sensors at the same height and installed at 45º. The deployment angle had no systematic effect on LWD measurements for the local tropical conditions, since the correlations between flat plate and elevated cylinder measurements were very high (R² > 0.91), which differed from the results obtained under temperate climatic conditions, where LWD measured by cylinders were two hours longer than by flat plate sensors.

Medidas da duração do período de molhamento (DPM) são necessárias para diversos sistemas de alerta fitossanitário, uma vez que a DPM é uma variável importante para a epidemiologia. O sensor cilíndrico é um sensor eletrônico barato e que pode ser facilmente produzido. Esse sensor foi inicialmente projetado para medir DPM em cebola, contudo alguns estudos indicam que ele também pode ser utilizado em estações meteorológicas e outras culturas. Portanto, o objetivo deste estudo foi avaliar o desempenho de sensores cilíndricos sob condições tropicais, no Brasil, tendo-se as medidas obtidas pelo sensor de placa como referência. Antes de serem instalados no campo, todos os sensors eletrônicos utilizados no estudo (placa e cilíndricos) foram pintados com tinta latex branca e, em seguida, submetidos a tratamento térmico. Testes de laboratório foram realizados para determinar um limiar de resistência, a partir do qual os sensores seriam considerados molhados, e o tempo de reposta dos sensores ao molhamento. No campo, todos os sensores foram inicialmente instalados na horizontal a 30 cm de altura sobre gramado, visando a avaliar a variabilidade entre os sensores de DPM. A variabilidade entre os sensores cilíndricos na horizontal foi reduzida com a adoção de um limiar de resistência específico para cada sensor. O coeficiente médio de variação para DPM diária medida pelos sensores cilíndricos foi igual a 9,7%. Posteriormente, os sensores cilíndricos foram instalados em 5 ângulos diferentes com a horizontal: 0, 15, 30, 45, and 60º. As medidas obtidas por esses sensores são comparáveis com a medida padrão, fornecida por sensores de placa instalados a 45º e mesma altura. A mudança do ângulo de instação não teve efeito sistemático sobre as medidas da DPM para as condições locais, uma vez que as correlações entre as medidas dos sensores de placa e sensores cilídricos inclinados foram bastante elevadas (R² > 0,91), o que difere dos resultados obtidos em condições de clima temperado, onde a DPM medida pelo sensor cilíndrico foi duas horas mais longa do que a medida pelo sensor de placa.

Performance of cylindrical leaf wetness duration sensors in a tropical climate condition

Leaf wetness duration (LWD) measurements are required for disease warning in several agricultural systems, since it is an important variable for the diagnose of plant disease epidemiology. The cylindrical sensor is an inexpensive and simple electronic LWD sensor initially designed to measure this variable for onions, however some studies show that it may be helpful for standard measurements in weather stations and also for different crops. Therefore, the objective of this study was to assess their performance under tropical climate conditions, in Brazil, having as standard measurements those obtained by flat plate sensors, which have presented very good performance when compared with visual observations. Before field assessments, all LWD sensors used in our study (flat plates and cylinders) were white latex painted and submitted to a heat treatment. Laboratory tests were performed in order to determine the resistance threshold for the sensor to be considered wet and the time response of the sensors to wetness. In the field, all cylindrical sensors were initially deployed horizontally 30-cm above a turfgrass surface in order to assess the variability among them with respect to LWD measurements. The variability among the horizontal cylindrical sensors was reduced by using a specific resistance threshold for each sensor. The mean coefficient of variation (CV) of LWD data measured by the cylindrical sensors was 9.7%. After that, the cylindrical sensors were deployed at five different angles: 0º, 15º, 30º, 45º, and 60º. Data of measurements made at these angles were compared with the standard measurement, obtained by flat plate sensors at the same height and installed at 45º. The deployment angle had no systematic effect on LWD measurements for the local tropical conditions, since the correlations between flat plate and elevated cylinder measurements were very high (R² > 0.91), which differed from the results obtained under temperate climatic conditions, where LWD measured by cylinders were two hours longer than by flat plate sensors.

Medidas da duração do período de molhamento (DPM) são necessárias para diversos sistemas de alerta fitossanitário, uma vez que a DPM é uma variável importante para a epidemiologia. O sensor cilíndrico é um sensor eletrônico barato e que pode ser facilmente produzido. Esse sensor foi inicialmente projetado para medir DPM em cebola, contudo alguns estudos indicam que ele também pode ser utilizado em estações meteorológicas e outras culturas. Portanto, o objetivo deste estudo foi avaliar o desempenho de sensores cilíndricos sob condições tropicais, no Brasil, tendo-se as medidas obtidas pelo sensor de placa como referência. Antes de serem instalados no campo, todos os sensors eletrônicos utilizados no estudo (placa e cilíndricos) foram pintados com tinta latex branca e, em seguida, submetidos a tratamento térmico. Testes de laboratório foram realizados para determinar um limiar de resistência, a partir do qual os sensores seriam considerados molhados, e o tempo de reposta dos sensores ao molhamento. No campo, todos os sensores foram inicialmente instalados na horizontal a 30 cm de altura sobre gramado, visando a avaliar a variabilidade entre os sensores de DPM. A variabilidade entre os sensores cilíndricos na horizontal foi reduzida com a adoção de um limiar de resistência específico para cada sensor. O coeficiente médio de variação para DPM diária medida pelos sensores cilíndricos foi igual a 9,7%. Posteriormente, os sensores cilíndricos foram instalados em 5 ângulos diferentes com a horizontal: 0, 15, 30, 45, and 60º. As medidas obtidas por esses sensores são comparáveis com a medida padrão, fornecida por sensores de placa instalados a 45º e mesma altura. A mudança do ângulo de instação não teve efeito sistemático sobre as medidas da DPM para as condições locais, uma vez que as correlações entre as medidas dos sensores de placa e sensores cilídricos inclinados foram bastante elevadas (R² > 0,91), o que difere dos resultados obtidos em condições de clima temperado, onde a DPM medida pelo sensor cilíndrico foi duas horas mais longa do que a medida pelo sensor de placa.

Agrometeorology and plant disease management: a happy marriage

Many plant disease outbreaks are triggered by suitably warm temperatures during periods of leaf wetness. Measurements or estimations of leaf wetness duration provided by Agrometeorologists have allowed Plant Pathologists to devise weather timed spray schemes which often reduce the number of sprays required to control plant diseases, thus lowering costs and benefitting the environment. In the near future, tools such as numerical weather models with small grid spacings, and improved weather radar, are expected to reduce the need for tight networks of surface observations. The weather models will also provide growers with forecast warnings of potential upcoming disease outbreaks, which will further enhance the contribution of agrometeorology to plant disease management.

A disseminação de muitas doenças de plantas é influenciada por condições favoráveis de temperatura durante o período de molhamento foliar. As medidas e estimativas da duração do período de molhamento foliar fornecidas pelos Agrometeorologistas têm permitido aos Fitopatologistas dar alertas sobre a necessidade de pulverizações com base nas condições meteorológicas, o que normalmente reduz o número de aplicações para o controle de doenças, resultando em menor custo de produção e menor contaminação do ambiente. Em um futuro próximo, ferramentas como os modelos numéricos de tempo, com alta resolução espacial, e os radares meteorológicos mais avançados, deverão reduzir a necessidade de redes de observação meteorológica de superfície mais densas. Os modelos meteorológicos também possibilitarão a previsão de disseminação potencial das doenças de plantas, o que irá aumentar ainda mais a contribuição da agrometeorologia para o controle fitossanitário mais racional.