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The cucurbit vegetable chieh-qua (Benincasa hispida var. chieh-qua How) is an important crop in South China and southeast Asian countries. Viral diseases cause substantial loss of chieh-qua yield. To identify the viruses that affect chieh-qua in China, ribosomal RNA-depleted total RNA sequencing was performed using chieh-qua leaf samples with typical viral symptoms. The virome of chieh-qua comprises four known viruses (melon yellow spot virus (MYSV), cucurbit chlorotic yellows virus (CCYV), papaya ringspot virus (PRSV) and watermelon silver mottle virus (WSMoV) and two novel viruses: cucurbit chlorotic virus (CuCV) in the genus Crinivirus and chieh-qua endornavirus (CqEV) in the genus Alphaendornavirus. The complete genomes of the two novel viruses in chieh-qua and three other isolates of CuCV in pumpkin, watermelon and cucumber were determined and the recombination signals of pumpkin and watermelon isolates of CuCV were detected. A reverse transcriptase PCR indicated that the dominant viruses of chieh-qua in Hainan are MYSV (66.67%) and CCYV (55.56%), followed by CuCV (27.41%), WSMoV (7.41%), cucumber mosaic virus (8.15%), zucchini yellow mosaic virus (6.67%), PRSV (6.67%) and CqEV (35.56%). Our findings support diagnostic and prevalence studies of viruses infecting chieh-qua in China, enabling sustainable control strategies for cucurbit viruses worldwide. [ABSTRACT FROM AUTHOR]

Powdery mildew, caused by Oidium heveae, is a major threat to rubber plantations worldwide. Population studies are scarce for this pathogen due to the lack of polymorphic molecular markers. In this study, sixteen polymorphic microsatellite loci were identified using DNA extracted from single lesions based on the whole-genome sequences of the pathogen. Primers of these loci were applied to 138 O. heveae samples from five counties in Hainan, China. The number of alleles per locus ranged from 2 to 11 and the gene diversity varied from 0.014 to 0.728. In total, 119 multilocus genotypes (MLGs) were observed for the 138 samples. However, only one sample was significantly distinct from the other samples base on DAPC analysis. Further study indicated that there were three subclusters for the other 137 samples, but no evidence for regional genotypic subdivision was identified. The microsatellite markers developed are very useful to study the genetic structure and the dispersal route of O. heveae, especially as it can use DNA extracted directly from infected leaves. [ABSTRACT FROM AUTHOR]

Areca palm, rubber tree, and coffee are always planted adjacent to each other or intercropped with each other, and Colletotrichum siamense was found to be the dominant species of Colletotrichum from these crops in Hainan, China. To study the population structure, pathogenicity, and fungicide sensitivity of C. siamense from these three crops in Hainan, isolates were first identified by their morphological characteristics and multilocus phylogeny. Then both permutational multivariate analysis of variance (PERMANOVA) and pairwise FST analysis showed that the original host and geographical origin (counties) had significant effects on genetic variation in the C. siamense population, explaining 5.6% and 16.3% of genetic variation, respectively. There were significant genetic differentiations between coffee and rubber tree populations but the genetic differentiation was small (FST = 0.04), while significant differentiations were observed among all populations from different counties except those between Haikou and Chengmai. Pathogenicity analysis using artificial inoculation showed that isolates had significantly higher disease incidence and diseased lesion diameter on wounded leaves than on nonwounded leaves. However, the original host of an isolate and geographical origin did not significantly affect the pathogenicity of the C. siamense populations from these three hosts. In vitro tests showed that there were no significant differences in the sensitivity of C. siamense populations to carbendazim, prochloraz, difenoconazole, and propiconazole from different original hosts and geographical origins. The present study suggests that host specialization of C. siamense has not occurred yet in coffee, rubber tree, and areca palm. [ABSTRACT FROM AUTHOR]

Black pepper (Piper nigrum L.) is one of the most ancient spice crops in the world. Hainan Province is the largest black pepper‐producing region in China. Viral disease is one of the main factors affecting black pepper. However, there are rarely studies on black pepper viral disease in China. We performed a survey in five major black pepper‐growing counties (Qionghai, Wenchang, Wanning, Haikou and Ding'an) of Hainan Province and collected 589 samples, with virus‐like symptoms from 2014 to 2019. Various viruses were identified including cucumber mosaic virus (CMV), piper yellow mottle virus (PYMoV), Piper DNA virus 1 (PDV‐1) and Piper DNA virus 2 (PDV‐2) using small RNA deep sequencing. The RT‐PCR screening of field samples indicated three types of infection: CMV alone (5.56%), PYMoV + PDV‐1 + PDV‐2 (25.00%) and PYMoV + CMV +PDV‐1 + PDV‐2 (69.44%). The data suggested that PYMoV might be the main pathogen causing virus diseases of black pepper. Two complete genome sequences of PYMoV were obtained. Phylogenetic analysis showed that PYMoV isolates from China were placed in a branch distinct from PYMoV isolates from India. To our knowledge, this is the first report of PYMoV, PDV‐1 and PDV‐2 naturally infecting black pepper, and the first genome sequence of PYMoV in China. [ABSTRACT FROM AUTHOR]

Powdery mildew is an important disease of rubber trees worldwide. To assess the effects of temperature and leaf age on conidial germination and disease development, conidia were inoculated onto rubber tree seedlings with leaves at three phenological stages (copper bronze, colour‐changing, and light green) and then incubated at six constant temperatures (10, 15, 20, 25, 30, and 35°C). Leaf age did not affect conidial germination (p =.296) whilst temperature did (p <.0001), although conidia were able to germinate at all tested temperatures. The estimated optimal temperature for conidial germination was 23.2°C. Leaf age, temperature, and their interactions had significant effects on conidial infection and hypha number (p <.0001). At 10 and 35°C, more than 2 and 4 days were needed for infection to complete, respectively, compared to <2, 1, 0.5, and 0.5 days for 15, 20, 25, and 35°C, respectively. Sporulation and mildew symptoms were only observed on those inoculated leaves of all stages at 20 and 25°C, and at the copper bronze stage only at 15°C. The latent period on the copper bronze leaves at 15°C was longer (9 days) than at 20 and 25°C (4 days). The latent period at 20 and 25°C increased from 4 to 7 days as the leaf development stage increased from copper bronze to light green. Therefore, temperature affected germination and postgermination growth of rubber tree powdery mildew, whereas leaf age primarily affected postgermination growth of the pathogen. [ABSTRACT FROM AUTHOR]

A disease on parthenium weed ( Parthenium hysterophorus L.) was observed in June 2008 in Danzhou of Hainan Province. Infected weeds showed phytoplasma-like associated symptoms such as severe stunting, excessive proliferation of shoots, inflorescence-clustering, green petal, small leaves and witches'-broom. The original cause of phytoplasma was further confirmed by polymerase chain reaction (PCR). PCR products of 1.8 kb were obtained using the universal primers pair (P1/P7) designed to amplify the entire 16S rDNA and the 16/23S intergenic spacer region in a direct PCR assay. The primers pair R16F2n/R2 was used to amplify a PCR product of 1.2 kb. Restriction fragment length polymorphism (RFLP) was used to analyze the partial 16S rDNA sequences (1.2 kb) of all phytoplasma DNA digested with five endonucleases ( Kpn I, Hpa II, Taq I, Rsa I, EcoR I). The RFLP patterns of the strain were found to be identical with that of the reference peanut witches'-broom phytoplasma. Based on the RFLP data, it is suggested that the phytoplasma strain belongs to subgroup 16SrII-A. This is the first demonstration of a 16SrII-A group phytoplasma associated with parthenium weed. [ABSTRACT FROM AUTHOR]

In July 2008, Stylosanthes guianensis with witches'-broom symptoms and Stylosanthes capitata with yellows symptoms were observed in Hainan Province, China. From the symptomatic S. guianensis, the phytoplasma-specific 16S rRNA gene fragment of 1.4 kb was amplified by a direct PCR, and a fragment of 16S rRNA gene of 1.2 kb was obtained from the symptomatic Stylosanthes capitata by a nested PCR. Sequencing results and NCBI BLASTn analysis showed that the two phytoplasmas belonged to group 16SrII. Polygenetic analysis and virtual RFLP classified the two phytoplasmas into subgroup 16SrII-A. [ABSTRACT FROM AUTHOR]