First report of papaya ring spot virus (PRSV) infecting jute (Corchorus olitorius) in India

Jute is the most important bast fibre crop of the world, which is mainly cultivated in India, Nepal, Bangladesh, China, Indonesia and South American countries. The fibre is utilized for making apparels, ropes, bags, carpets etc (Biswas et al. 2014). This bio-fibre is gaining importance due to growing environmental consciousness worldwide. In June 2019, we noticed jute plants (less than 2%) showing virus like symptoms viz., downward curling, puckering, angular brownish to yellowish spots etc in a farmer’s field at Amdanga Block, North 24 Paraganas, West Bengal, India. To identify the virus, five symptomatic leaves from five different plants were used for high throughput sequencing (HTS). We extracted total RNA from each leaf which was subjected to construction of cDNA libraries. Sequencing was done on Illumina Hiseq 4000 (CytoScan, Thermo Fisher). Approximately 46 million 105 nt paired end reads were generated. Raw reads were trimmed and filtered to perform de novo assembly as described previously by (Grabherr et al. 2013). The obtained contig was 10,326 bp nucleotides (nt) long and in BLASTn against GenBank showed highest identity with papaya ring spot virus (PRSV) with the contig covering 99.6% of the viral genome. The obtained contig shared 99.33% sequence similarity with PRSV strain P (Accession No. MT470188). The selected leaf samples were also tested by double-antibody sandwich (DAS)- enzyme linked immunosorbent assay (ELISA) for papaya ring spot virus (PRSV) along with some common viruses, viz., Potato leaf roll virus (PLRV), Watermelon mosaic virus, Cowpea mosaic virus and Cucumber mosaic virus with the help of commercial diagnostic kits (Agdia). However, only the test with PRSV gave positive reaction for the symptomatic samples. The major symptoms of PRSV on papaya are severe mosaic, chlorosis, reduced lamina with curling and puckering (Gonsalves et al. 2010). To confirm PRSV infection, five symptomatic leaf samples (used for HTS) were collected and whole RNA was extracted from the samples using RNeasy plant minikit (Qiagen, USA). Reverse transcriptase polymerase chain reaction (RT-PCR) was conducted by using isolated RNA. One pair of PRSV specific primer (PSRV1F: 5' TTAAATCTGATTCGTC 3' PRSV 1R: 5'GAAATTCACGCAAAGTCGA3') was developed by using primer BLAST software and was used in RT-PCR assays. Amplified fragments were cloned and sequenced and all the fragments shared 98% sequence identity with PRSV. One of the amplicons was deposited in NCBI (Accession No. MN615832). Crude sap was prepared by homogenizing PRSV-infected jute leaf tissues in 0.1 M sodium phosphate buffer and 2% carborundum dust was added as abrasive (Holkar et al. 2018). The sap was then gently rubbed on to the healthy papaya leaves for inoculation. Typical PRSV like symptoms appeared in inoculated leaves 10 days post inoculation which confirmed the presence of PRSV-P. PRSV was detected by RT-PCR as well as (DAS)-ELISA from all inoculated infected papaya leaf tissues, but could not be detected from uninoculated healthy papaya tissues. To the best of our knowledge, this is the first report of PRSV-P infecting jute in India. References: Biswas, C. et al. 2014. Plant Dis. 98(4): 565. https:// doi.org/10.1094/PDIS-08-13-0826-PDN. Gonsalves, D. et al. 2010. The Plant Health Instructor. https:// doi.org/10.1094/PHI-I-2010-1004-01 Holkar, S. K. et al. 2018. Crop Protection. 108:110-119. https://doi.org/10.1016/j.cropro.2017.12.013 Grabherr, M. et al. 2011. Nat Biotechnol. 29(7): 644-652.

Influence of Bioregulators on Latex Yield and Proteolytic Activity of Papain Extracted from Field-grown TNAU Papaya CO. 8 under the Natural Incidence of Papaya Ring Spot Virus

The cultivation of papaya for both fruits and commercial papain extraction drastically suffers often due to the devastating incidence of papaya ringspot virus disease. The present study aimed to find out whether the papain yield and quality are influenced when bioregulators are applied to manage papaya ring spot virus (PRSV) incidence or as a measure to limit the height of the crop in TNAU Papaya CO.8 variety grown in the open field with the natural incidence of PRSV. Three growth retardants [250 ppm of prohexadione calcium (G2), 500 ppm of mepiquat chloride (G3) and 500 ppm of chlormequat chloride (G4)] and three stress tolerance promoting bioregulators [100 ppm of methyl jasmonate (S2), 100 ppm of salicylic acid (S3) and 0.6 g/ l of strobilurins (S4)] were employed in the study. Water spray (G1 and S1) was kept as control. The treatments were imposed at 4th and 8th months after planting. The growth retardants were initially sprayed and, in the subsequentweek ,the stress tolerance promoting bioregulators were sprayed as per the recommended doses. Among the combinations tried, spraying 250 ppm of prohexadione calcium followed by water spray (G2S1) a week later recorded the lowest plant height (140.79 cm) at the time of fruiting. The treatment combination G 2S2 involving 250 ppm of prohexadione calcium and 100 ppm of methyl jasmonate registered a higher number of fruits (42.17). The combination of 250 ppm of prohexadione calcium along with 100 ppm of salicylic acid (G2S3) was found to be the most effective combination to manage PRSV without compromising papain production and to enhance the proteolytic activity significantly (44378.36 TU g-1) as compared to other combinations. The treatment combination of 500 ppm of mepiquat chloride and 100 ppm of salicylic acid (G3S3) favored higher dry latex yield apart from a significant reduction in PRSV symptoms. Both these treatment combinations (G2S3 and G 3S3) merit due consideration in PRSV management for commercial fruit and papain production in papaya

Correlation of Leaf Parameters with Incidence of Papaya Ring Spot Virus in Cultivated Papaya and its Wild Relatives

Papaya ring spot virus (PRSV) disease has been the major impediment in papaya cultivation. The disease is transmitted through three aphid vectors and field tolerance towards this disease varies among Carica papaya cultivars as well as within the Vasconcellea genus. Leaf morphological traits are known to have influence on the probing preferences of aphids. Hence, this study was conducted to know whether the leaf parameters could contribute to the incidence of PRSV possibly by influencing the probing or feeding behaviour of aphid vectors. Leaf parameters viz., leaf thickness, leaf epicuticular wax content, presence and type of trichomes, trichome density were correlated with disease incidence at field conditions. The result revealed that leaf thickness along with epicuticular wax content had significant negative correlation with disease incidence. Similarly, trichome density had negative impact on disease incidence at 99.92% significance level. High epicuticular wax content and high trichome density in V. cauliflora and V. cundinamarcensis were found to be negatively associated with low to very low infection indicating that these parameters may have limited the vector transmission significantly.

Efecto de inhibidores en el control del Papaya ringspot virus y productividad en papaya (Carica papaya)

La papaya (<em>Carica papaya</em>) es un frutal muy aceptado en el mercado de exportación por generar alta rentabilidad, en México se cultiva en zonas tropicales y subtropicales. El <em>Papaya ring spot virus</em> (PRSV) ocasiona la enfermedad más restrictiva de la producción de este cultivo en México y el mundo, por lo cual se evaluó la eficacia de Inhibitovir® y Virus Stop® en campo a 3 mL L-1 y 6 mL L-1 sobre incidencia, intensidad y control de la virosis, altura de planta, diámetro de tallo, número de flores y frutos, peso del fruto y rendimiento. El PRSV se detectó hasta el quinto mes después del trasplante, tiempo para que la plantación lograra un buen amarre de frutos, sin embargo, la eficacia de los inhibidores virales no tuvo diferencias significativas sobre plantas testigo infectadas con el PRSV, así como en altura de planta, diámetro de tallo, número de flores y frutos, peso del fruto y rendimiento.