Publications

4560

Areca palm yellow leaf phytoplasma (AYLP) and Areca palm velarivirus 1 (APV1) are two critical pathogens associated with yellow leaf in areca palm, resulting in devastating damage to areca production. However, evidence of their coinfection remains unclear. Areca palms showing yellow leaf symptoms (suspected to be caused by phytoplasma and velarivirus 1) were surveyed and sampled around Hainan Island of China, and the pathogens were identified and analyzed in the study. The results showed that infectious leaf yellowing symptoms of areca palms primarily occurred in eastern and southern cities and counties of Hainan Island. Molecular identification revealed coinfection of ‘Candidatus Phytoplasma asteris’ (16SrI-B subgroup) and APV1 in individual areca palm samples. Among 520 samples tested by qPCR, AYLP was detected in 23.46% (n = 122), APV1 in 53.46% (n = 278), and a coinfection with both AYLP and APV1 in 10.96% (n = 57). A statistically significant difference in APV1 detection rate was observed between asymptomatic and symptomatic plantations (P < 0.0001), suggesting that APV1 infection likely represents an important risk factor for yellowing development in cultivated areca palm populations. These results confirm the coinfection of AYLP and APV1 on Hainan Island, offering epidemiological evidence for targeted disease management.

Persian lime (Citrus latifolia Tan.) is a crop of high economic and social relevance in Mexico due to its nutritional properties, associated health benefits, and particularly the foreign exchange generated through exportation. In recent years, the state of Veracruz has become the leading production region for this species at the national level. However, this crop is susceptible to several vascular diseases that cause significant economic losses in the citrus industry. Among these, Huanglongbing (HLB) is considered the most destructive disease, exerting the greatest impact on Persian lime production. This study involved the collection of Persian lime leaf tissues from different sources: field-grown plants, certified nursery plants, and plants previously sanitized through micrografting and thermotherapy processes. Genomic DNA was extracted from the collected samples. DNA integrity was verified by agarose gel electrophoresis and quantified using a NanoDrop One-C spectrophotometer. Quantitative Real-Time Polymerase PCR (qPCR) was performed using a QuantStudio One thermocycler and SYBR Green Master Mix as the fluorescent detection system. Samples were considered HLB-positive when the cycle threshold (Ct) value was below 35 during the quantification of Candidatus Liberibacter asiaticus. The results support the early, precise, and efficient detection of CLas by qPCR in Persian lime plants, thereby contributing to improved diagnostic and phytosanitary management strategies for this crop.

Shorea assamica is an evergreen to semi-evergreen tree in the family Dipterocarpaceae. It is widely distributed in China, India, Myanmar, Malaysia, Indonesia, the Philippines, and other regions. In China, it is found in western Yunnan and southeastern Tibet. The species is valued for its aromatic white resin, which serves as an important chemical raw material (Ang and Maruyama 1997). In May 2025, S. assamica trees exhibiting symptoms of witches'-broom disease and plexus bud disease were found at Mengmao, WanDing, Nongdao, Jiegao, and Jiexiang in RuiLi County (97°31′-98°02′ E, 23°38′-24°14′ N), Dehong Prefecture, Yunnan Province, China. Following disease onset, the overall growth of infected plants was significantly slower compared to healthy individuals. Infected plants exhibited excessive branching, shortened internodes, and a clustered growth symptom. Lateral buds on diseased tree frequently developed into abnormal twigs, with many buds forming dense clusters and only a few producing leaves. The disease was designated as S. assamica witches'-broom (SAWB). Across the seven surveyed areas, over 74% of trees displayed symptoms, while the disease incidence in nursery-cultivated seedlings was 66%. The disease was found to be spreading actively. A total of 56 samples were collected from seven areas, including 42 symptomatic plants and 14 asymptomatic plants. The lateral stem tissues of 30 samples were observed under a scanning electron microscope (Hitachi S-3000N). Spherical bodies were found in the phloem sieve cells of symptomatic plants. Total DNA was extracted from 56 plant samples using the CTAB method (Porebski et al. 1997) and subjected to nested PCR testing. Double-distilled water was used as negative control, and the DNA extracted from Dodonaea viscosa affected by D. viscosa witches’-broom disease was used as a positive control. Nested PCR was employed to amplify the pathogen’s 16S rRNA gene (Lee et al. 1993; Schneider et al. 1993), and 1.2 kb segment was produced (GenBank accessions: PZ025091, PZ025092, and PZ025093). PCR specific to the ribosomal protein (rp) gene yielded a segment of approximately 1.2 kb (Lee et al. 2003) (GenBank accessions: PZ028767, PZ028768, and PZ028769). The fragment size from 31 samples was consistent with the positive control, confirming the association of phytoplasma with the disease. A BLAST analysis of the 16S rRNA sequences of S. assamica witches’-broom phytoplasma showed that it shared 99.68% identity with that of Rice orange leaf phytoplasma (GenBank accession: JQ965690). The rp sequence shared 99.83% identity with that of Salix tetradenia witches'-broom phytoplasma (GenBank accession: KC117314). An analysis with iPhyClassifier (Zhao et al. 2013) showed that the virtual RFLP pattern derived from the 16S rDNA fragment of SAWB phytoplasma was identical (similarity coefficient 0.98) to the reference pattern of 16Sr group I, subgroup B (OY-M, GenBank accession AP006628). The SAWB phytoplasma is a variant of 16SrI-B. The phylogenetic tree was reconstructed based on 16S rRNA and rp gene sequences using MEGA (Tamura et al. 2013). The analysis was conducted using the neighbor-joining method with 1,000 replicates of bootstrap analysis. The results indicated that the SAWB phytoplasmas grouped into clades including phytoplasmas belonging to 16SrI-B and rpI-B, respectively. In addition, 1-year-old S. assamica were used for grafting assays in nursery, the twigs from infected S. assamica under natural conditions were used as a scion, and the phytoplasma was detected using nested PCR after grafting for 40 days. To the best of our knowledge, S. assamica is a new host of ‘Ca. P. asteris’-related strain (16SrI-B) in China. The newly emerged disease is a threat to S. assamica.