Detecting Phytophthora cinnamomi associated with dieback disease on Carya cathayensis using loop-mediated isothermal amplification

Chinese hickory (Carya cathayensis Sarg.) is an economically and ecologically important nut plant in China. Dieback and basal stem necrosis have been observed in the plants since 2016, and its recent spread has significantly affected plant growth and nut production. Therefore, a survey was conducted to evaluate the disease incidence at five sites in Linan County, China. The highest incidence was recorded at the Tuankou site at up to 11.39% in 2019. The oomycete, Phytophthora cinnamomi, was isolated from symptomatic plant tissue and plantation soil using baiting and selective media-based detection methods and identified. Artificial infection with the representative P. cinnamomi ST402 isolate produced vertically elongated discolorations in the outer xylem and necrotic symptoms in C. cathayensis seedlings in a greenhouse trial. Molecular detections based on loop-mediated isothermal amplification (LAMP) specific to P. cinnamomi ST402 were conducted. Result indicated that LAMP detection showed a high coherence level with the baiting assays for P. cinnamomi detection in the field. This study provides the evidence of existence of high-pathogenic P. cinnamomi in the C. cathayensis plantation soil in China and the insights into a convenient tool developed for conducting field monitoring of this aggressive pathogen.

Detecting Phytophthora cinnamomi associated with dieback disease on Carya cathayensis using loop-mediated isothermal amplification

Chinese hickory (Carya cathayensis Sarg.) is an economically and ecologically important nut plant in China. Dieback and basal stem necrosis have been observed in the plants since 2016, and its recent spread has significantly affected plant growth and nut production. Therefore, a survey was conducted to evaluate the disease incidence at five sites in Linan County, China. The highest incidence was recorded at the Tuankou site at up to 11.39% in 2019. The oomycete, Phytophthora cinnamomi, was isolated from symptomatic plant tissue and plantation soil using baiting and selective media-based detection methods and identified. Artificial infection with the representative P. cinnamomi ST402 isolate produced vertically elongated discolorations in the outer xylem and necrotic symptoms in C. cathayensis seedlings in a greenhouse trial. Molecular detections based on loop-mediated isothermal amplification (LAMP) specific to P. cinnamomi ST402 were conducted. Result indicated that LAMP detection showed a high coherence level with the baiting assays for P. cinnamomi detection in the field. This study provides the evidence of existence of high-pathogenic P. cinnamomi in the C. cathayensis plantation soil in China and the insights into a convenient tool developed for conducting field monitoring of this aggressive pathogen.

Prevalence and Molecular Characterization Coat Protein Gene of Tobacco streak virus Causing Peanut Stem Necrosis Disease in Coastal and Rayalaseema Districts of Andhra Pradesh, South-India

Background: Peanut stem necrosis disease (PSND) caused by Tobacco streak virus (TSV) is a major constraint for groundnut production in Andhra Pradesh (A.P.). However, studies on prevalence and spread of the disease confined to only few districts of A.P. with this background current study focused on incidence and spread of the disease in entire state of A.P. Further an isolate of TSV occurring in A.P. characterized on the basis of genetic features by comparing with other TSV isolates originated from different hosts and locations from world.Methods: Roving survey was conducted during kharif 2017-18 in groundnut growing districts of Andhra Pradesh (A.P.) for peanut stem necrosis disease incidence. Groundnut plants showing PSND symptoms were collected and tested with direct antigen coating enzyme linked immunosorbent assay (DAC-ELISA). Groundnut samples found positive by ELISA once again tested by reverse transcription polymerase chain reaction (RT-PCR). The representative TSV-GN-INDVP groundnut isolate from Prakasham district was maintained on cowpea seedlings by standard sap inoculation method in glasshouse for further molecular characterization. The Phylogenetic tree for coat protein (CP) gene was constructed using aligned sequences with 1000 bootstrap replicates following neighbor-joining phylogeny.Result: Thirty-eight (52.7%) of seventy-two groundnut samples collected from different locations in A.P were given positive reaction to TSV by DAC-ELISA. For the first time, PSND incidence observed in coastal districts (Krishna, Guntur, Sri Pottisriramulu Nellore, Prakasham) of A.P. Maximum PSND incidence recorded from Bathalapalli (22.2%) and the minimum incidence in Mulakalacheruvu (4.1%). The coat protein (CP) gene of TSV-GN-INDVP groundnut isolate was amplified by RT-PCR and it shared maximum per cent nucleotide identity (97.51-98.62%) with TSV isolates from groundnut and other different crops reported in India. All Indian isolates cluster together irrespective of crop and location based on the phylogenetic analysis.

First Report of Choanephora cucurbitarum Causing Leaf Wilt, Flower Rot, and Stem Necrosis on Crotalaria breviflora

Crotalaria breviflora (Fabaceae) is used as green manure crop because of its nitrogen fixation and nematode control (Nascimento et al. 2020). In April 2018, leaf wilting, flower rot, and stem necrosis symptoms were observed on C. breviflora with 100% incidence, in Sorriso (12° 33′ 31″ S, 55º 42′ 51″ W), Santa Carmem (11° 55′ 52″ S, 55º 16′ 47″ W), and Sapezal (12º 59′ 22″ S, 58º 45′ 52″ W) counties in the state of Mato Grosso, Brazil. Three monosporic isolates were isolated from symptomatic leaves, cultivated in potato dextrose agar (PDA) medium, and deposited at the Cultures Collection of the University of Brasilia (codes CCUB 1293, CCUB 1667, CCUB 1668). Colonies on PDA were white and cottony with presence of hyaline and coenocytic hyphae. The mycelia later became pale yellow with abundant reproductive structures. Sporangiophores were hyaline, aseptate, unbranched, and apically dilated to form a clavate vesicle, which produced secondary vesicles bearing sporangiola. Secondary vesicles were clavate, light brown, and 37 to 51 µm in diameter. Sporangia were brown to dark brown, globular to ellipsoid, 115 to 140 µm long, and 96 to 122 µm wide. Sporangiospores (n=30) were brown to reddish-brown, ellipsoid to ovoid, with longitudinal striae, 14 to 19 µm long, and 8 to 12 µm wide. Some with hyaline appendages at both ends. Their morphological characteristics were consistent with the descriptions of Choanephora cucurbitarum (Kirk 1984). To confirm the identity, the DNA of the three isolates was extracted and the sequences of Small Subunit (SSU), Large Subunit (LSU), and complete Internal Transcribed Spacer (ITS) of rDNA were amplified using V9G, ITS3, and LR5 primers (GenBank acc. no: MN897836, MN897837 and MN897838). The sequences were aligned with the MAFFT software. The alignment matrix was subjected to Maximum Likelihood (ML) analysis using RAxML v. 8 and Bayesian Inference performed in MrBayes v.3.1.2. The tree was edited in the FigTree software. The sequences showed 100% identity with the sequences from C. cucurbitarum found on the GenBank. To confirm pathogenicity, a suspension at 5.4 ×106 spores/ml was prepared from a 15-day-old culture grown at 25°C and sprayed on asymptomatic plants of C. breviflora. Sterilized water was sprayed as the control. Plants were kept in a humid chamber at 20°C for 48 h. Initial symptoms were visualized 16 days after inoculation. Complete necrosis of leaves and stems with spore mass on infected tissue was observed 19 days after inoculation. To satisfy the Koch’s postulates, the fungus was successfully reisolated from the infected tissues. No symptoms were observed on the control plants. In Brazil, this pathogen has been reported on Brassica oleracea var. capitata, Capsicum annuum, Crotalaria spectabilis, Cucurbita sp., and Vigna unguiculata (Alfenas et al. 2018; Mendes and Urben, 2019). C. cucurbitarum has been reported to have a wide range of hosts (Farr and Rossman, 2020). It can infect the crops grown in rotation or in succession, including common bean, corn, cotton, quinoa, soybean, and sunflower. Therefore, this pathogen is of epidemiological importance and poses a threat to the croplands where environmental conditions are conducive to the disease to develop and spread. To our knowledge, this is the first report of C. cucurbitarum causing leaf and flower wilt, and stem rot on C. breviflora in the world. Acknowledgment We thank the Environmental Sciences Graduate Program, Federal University of Mato Grosso, University of Brasilia, PROPeq/PROPG-UFMT, EMBRAPA, CODEX/UFMT, Institute of Agricultural and Environmental Sciences (ICAA)/UFMT and CAPES for providing the Master's scholarship. References Alfenas, R. F., et al. 2018. Plant Dis.102:1456. https://doi.org/10.1094/PDIS-10-17-1610-PDN, Google Scholar. Farr, D. F., and Rossman, A. Y. 2020. Fungal Databases, Syst. Mycol. Microbiol. Lab., ARS, USDA. Retrieved May 26, 2020 from https://nt.ars-grin.gov/fungaldatabases/, Google Scholar. Kirk, P. M. 1984. Mycol Paper. 152:1. Google Scholar. Mendes, M. A. S., and Urben, A. F. 2020. Fungos relatados em plantas no Brasil, Retrived May 26, 2020 from http://pragawall.cenargen.embrapa.br/aiqweb/michtml/fgbanco01.asp, Google Scholar. Nascimento, D. D. et al. 2020. Bioscience Journal. 36:713. https://doi.org/10.14393/BJ-v36n3a2020-42248, Google Scholar.

Effects of cowpea mild mottle virus on soybean cultivars in Brazil

Soybean stem necrosis is caused by cowpea mild mottle virus (CPMMV), transmitted by the whitefly Bemisia tabaci. CPMMV has already been recorded in all major soybean-producing areas of Brazil. The impacts caused by CPMMV to the current Brazilian soybean production are unknown, thus the main objective of this study was to evaluate the effects of CPMMV infection on the main important soybean cultivars grown in the Southern and Midwestern regions of Brazil. Although asymptomatic in some of the tested cultivars, CPMMV infection significantly reduced the plant height, the number of pods per plant and the 1,000-grain weight. In addition, estimated yield losses ranged from 174 to 638 kg ha−1, depending on the cultivar. Evidence of seed transmission of CPMMV was observed in the BMX POTÊNCIA RR cultivar. These results suggest that CPMMV could have an important role in the reduction of soybean productivity in Brazil, but symptomless infections might be hiding the actual impact of this pathogen in commercial fields and infected seeds could be the primary inoculum source of the virus in the field.

The Effect of Salinity on the Growth of Lavender Species

Long term degradation of water quality from natural resources has led to the use of alternative water resources for irrigation that are saline. Saline water irrigation in floriculture for the production of nursery crops requires an understanding of plant response. The pot growth of four lavender species (Lavandula angustifolia, Lavandula dentata var. dentata, Lavandula dentata var. candicans and Lavandula stoechas) irrigated with water containing different concentrations of NaCl (0, 25, 50, 100 and 200 mM) was investigated under greenhouse conditions. Overall results of different plant growth variables were consistent, showing a significant decrease at 100 and 200 mM NaCl. All lavender species showed signs of salinity stress that included chlorosis, followed by leaf and stem necrosis at NaCl concentrations greater than 50 mM. L. dentata var. dentata showed the greatest plant growth followed in descending order by L. dentata var. candicans, L. stoechas and L. angustifolia. Despite greater growth of L. dentata var. dentata, the appearance of L. dentata var. candicans was “healthier”. In areas with saline irrigation water, L. dentata var. dentata and L. dentata var. candicans are proposed for the production of lavender nursery crops.

Comparison of chili pepper breeding populations for agronomic traits and polygenic resistance to Phytophthora blight

ABSTRACT Belonging to the Oomycete class, Phytophthora capsici has wide range of host profile and is responsible for many devastating diseases in many countries. In addition to time consuming problem for transferring resistance to susceptible varieties, backcrossing method causes losing of genes providing resistance to susceptible varieties. In this study transferring P. capsici resistance genes to susceptible chili pepper lines was aimed during the extensive breeding period and resistant lines were confirmed by marker assistance. Two different breeding populations from CM334 and PM217 were compared by stem inoculation test to determine receptivity, inducibility, and stability resistance component. CM334 was found more effective for transferring all resistance components while PM217 was found suitable for keeping agronomic traits along with two important resistance component inducibility and stability. These two resistant components were found highly correlated to length of stem necrosis. C-29 and C-18 have been improved from CM334 as resistant as CM334; P-73 and P-77 have been improved satisfactorily resistant and yielded lines from PM217. Marker assisted selection proved that resistance of lines differentiated phenotypically despite the genotypes have the same genes.

Economical important phytopathogenic diseases in Vanilla planifolia: A review paper.

The vanilla orchid is one of the high value and most favorite culinary crops in the world. The aroma or flavor of vanillin make the plant as an important economic crop in the food and beverage industry. Nevertheless, the production of vanilla is hindering by two main factors which are the propagation and diseases. Devastating disease has cause losses in bean production as it can infect the vines at any times and at all stages of the plant growth. As a result, it is cause the losses of quality and quantity of pod production. Some of important diseases associated with fungal and viral on vanilla such as Fusarium root and stem rot, anthracnose, black rot, sclerotium rot, rust, mosaic and stem necrosis are discussed in this review paper.