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Foliar nematodes (Aphelenchoides spp.) are known to be parasites of tobacco in restricted areas, but symptoms caused by A. besseyi in tobacco are not well characterized, despite the great importance of this nematode worldwide. This study aimed to evaluate the host reaction of four Nicotiana tabacum cultivars (Comum, Xanthi, Samsun, and TNN) and N. benthamiana cultivar Comum to A. besseyi and to characterize the symptoms and the parasitism of this nematode. Two experiments were conducted under greenhouse conditions with controlled humidity and temperature, in which the plants were inoculated with 600 A. besseyi. At 30 days after inoculation (DAI), nematodes present in the soil, roots, and shoot parts were extracted, and roots and shoot tissues were stained with acid fuchsin. A high number of A. besseyi was obtained per gram of shoot tissues (125 to 2,169 nematodes), and severe symptoms were observed in leaves and inflorescences of all cultivars. The symptoms included foliar distortion and deformation, necrotic spots delimited by the veins, flower abortion, and poor development of plants. In addition, A. besseyi was observed to penetrate tobacco roots at 30 DAI, and nematodes were also observed in the foliar mesophyll, inflorescences, and stems, a parasitism that has not been previously reported in tobacco plants. The disease caused by A. besseyi in tobacco could be a concern for growers in southern and northeastern Brazil because this nematode can cause severe damage to the marketable leaves of tobacco, reducing its commercial value.

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Soybean (Glycine max L.) is produced in over 70,000 ha in the Altillanura Region, eastern Colombia (Agronet 2023). From 2018 to 2020, foliar symptoms like green stem and foliar retention of soybean, which in Brazil can cause up to 100% soybean yield losses (Meyer et al. 2017), were observed in soybean fields in Colombia. During 2020, samples from symptomatic plants in reproductive stages (R1-R8) were collected from different commercial soybean fields in the Altillanura Region. Over 200 samples were processed, using an incubation method described in Coyne et al. (2014). Nematodes were recovered from photosynthetic leaf tissues and enlarged nodes/buds with population densities ranging from 13 to 132 and 36 to 936 nematodes/10g, respectively. Adult females were morphologically and molecularly characterized as Aphelenchoides pseudobesseyi (Oliveira et al. 2019; Subbotin et al. 2020). Female body length (n = 20) ranged from 653.3 to 806.3 µm (mean = 723 µm ± 52.7), stylet length from 11.0 to 12.3 µm (11.8 µm ± 0.3), body diameter from 14.8 to 17.9 µm (16.3 µm ± 1.1), post-uterine sac length from 38.7 to 51.9 µm (44.6 µm ± 5.1), vulva to anus from 145.5 to 223.2 µm (172.2 µm ± 22.4), and 26% of the vulva-anus distance. Genomic DNA was extracted (QIAGEN DNeasy® Blood & Tissue kit) from a pool of nematodes. The D2A/D3B (Tenente et al. 2004) primers were used to amplify and sequence the D2/D3 expansion region of the 28S rRNA gene. PCR product (~759 bp) was purified, sequenced, deposited in GenBank (OQ930285), and compared to previously deposited sequences (e.g., KX356756, KY510840, KY510839, KY510841, KT692694, KY510842, MH187565) by means of the BLAST algorithm. Similarly, 988F and 18SR-Burs (De Jesus et al. 2016) primers were used to amplify and sequence the near full-length 18S RNA gene (SSU). PCR product was purified, sequenced, deposited in GenBank (OQ954344), and compared to previously deposited sequences (e.g., KT454962, KT943534, KT943535, KY510835, KY510836, KY510837, KY510838, MH187565). Phylogenetic Bayesian analysis (Ronquist et al. 2012) of the of the D2/D3 and 18S regions placed this nematode from Colombia in the A. pseudobesseyi clade (PP = 100). To fulfill a modified Koch's postulates, the A. pseudobesseyi population described above was used in a greenhouse assay. In total, 120 soybean plants (cv. Flor Blanca) were infected with 200 A. pseudobesseyi (females + males)/plant. Briefly, at cotyledon stage (VC), 50 µl aliquot containing 50 A. pseudobesseyi was delivered onto each cotyledon and unifoliolate leaves (200 nematodes/plant). Sterile water was delivered to 80 plants which served as control. Plants were kept in the greenhouse at approximately 25°C and covered with clear plastic bag for 72 h to maintain over 90% relative humidity. After 15, 30, 45, and 60 days, soybean plants (n = 20) were processed, A. pseudobesseyi quantified, and the average reproduction factor (final population/initial population) was 0.1, 2.9, 14.0, and 1.8, respectively. Infected plants showed symptoms of blistering leaves with malformation (midrib vein twist), and A. pseudobesseyi was not observed in control plants. To our knowledge, this is the first report of A. pseudobesseyi parasitizing soybean buds and leaves in Colombia. Soybean is an important commodity for the Altillanura Region, and it is important to monitor the risk posed by this nematode. Furthermore, a better understanding of the nematode-host interaction and epidemiology in Colombia soybean producing regions is needed.

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Several species of nematodes are known to cause losses to cowpea (Vigna unguiculata) throughout the world. In Brazil, Aphelenchoides besseyi was described as causing damage on soybean, cotton, and common bean, but no report was found about the parasitism of this nematode in cowpea. This study aimed to verify the host reaction of cowpea cultivars to A. besseyi. The experiment was conducted under greenhouse conditions, using two A. besseyi populations as inocula, obtained from symptomatic soybean and cotton plants collected in naturally infested fields. Cultivars Imponente, Aracê, Guariba, Tumucumaque, Nova Era, and Tracuateua were inoculated with 500 A. besseyi of each population, separately, into soil, and after 30 days from the inoculation nematodes were extracted from shoot systems. Both populations were able to parasitize all the cowpea cultivars. Independently of the cultivar, cowpea plants exhibited symptoms of leaf deformation similar to those described for soybean, cotton, and common bean and, in addition, severe brooming was observed, with the interior of the stems being porous and necrotic. To our knowledge, this is the first report of parasitism by A. besseyi of cowpea in Brazil, under greenhouse conditions, increasing the list of hosts of this nematode.

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Aphelenchoides pseudogoodeyi has recently been reported in association with seeds of forage grasses and rice in Brazil and senescent strawberry plants, in the United States. This nematode is likely a mycophagous species; however, so far, its pathogenicity potential to plants is unclear. This study aimed to verify the pathogenicity of A. pseudogoodeyi to two species of ornamental plants. The experiments were conducted by inoculating A. pseudogoodeyi onto Bird's-Nest Fern (Asplenium nidus) and Oriental Lily (Lilium speciosum) leaves, using two inoculation methods (with and without injury). After 40 days of inoculation (DAI) in Bird's-Nest Fern and 5, 10, 20 and 40 DAI in Oriental Lily, the pathogenicity and the host efficiency were evaluated by symptoms observation and by severity, final nematode population and reproductive factor (RF), respectively. Additionally, a histopathological study was performed by inoculating A. pseudogoodeyi onto Bird's-Nest Fern for observing anatomical alterations. A. pseudogoodeyi was able to cause local necrotic lesions on both Bird's-Nest Fern and Oriental Lily leaves. However, the presence of injury was essential to enable A. pseudogoodeyi to penetrate and cause those symptoms in both plant species. Also, the total population of A. pseudogoodeyi decreased drastically over time and RF was <1, which characterized these species as poor-host or resistant plants. A. pseudogoodeyi penetrated into the foliar tissue and induced a total destruction of the mesophyll and collapse of the cells, with the formation of large intercellular spaces. It is concluded that A. pseudogoodeyi is an opportunistic pathogen as injury was required to induce symptoms in Bird's-Nest Fern and Oriental Lily.

RESUMO

Aphelenchoides besseyi is the causal agent of soybean green stem and foliar retention syndrome known as "Soja Louca II." This nematode has recently been reported parasitizing cotton in Brazil. In Costa Rica, it causes the symptoms known as "amachamiento" and false angular spots in common bean (Phaseolus vulgaris). Due to the great importance of beans to Brazilian agriculture, the objective of this research was to study the pathogenicity of A. besseyi in common bean under greenhouse conditions, including its endoparasitic relationships by staining root and shoot system tissues with fuchsin acid. In addition, A. besseyi was collected and quantified from shoot systems 30 days after inoculation by washing the tissue in water and blender centrifugal flotation. We observed the symptoms of amachamiento, leaf and vein deformation in the expanded trifoliate leaves, and also leaves with necrotic, brown to reddish and angular lesions, characteristics from false angular spot, and deformed stems characterized by enlargement of nodes, retortions, and necrotic lesions. High numbers of nematodes were found inside common bean plants. This is the first report of the pathogenicity and symptoms caused by A. besseyi in common bean in Brazil. These findings are important for development of management strategies to avoid losses on bean cropped in infested areas.

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