Publications

4641

Abstract Candidatus Liberibacter asiaticus ( C Las), the causal agent of Huanglongbing, is transmitted by the Asian citrus psyllid ( Diaphorina citri ) in a persistent, propagative manner. Reproducible transmission studies require not only controlled inoculation assays but also standardized propagation of citrus host plants, maintenance of D. citri colonies, and reliable pathogen detection. This study presents a comprehensive set of coordinated protocols that collectively support controlled C Las transmission experimentation. The workflow includes propagation of Citrus macrophylla for colony and assay use; generation and maintenance of C Las‐infected citrus and D. citri colonies; synchronization of D. citri populations to enable C Las acquisition; and a controlled single‐ D. citri transmission assay with defined inoculation access periods, and a Dellaporta‐based nucleic acid extraction method compatible with citrus and psyllid tissues. Quantitative PCR detection is integrated throughout to verify infection status, determine C Las titer, and monitor C Las movement and systemic colonization in citrus over time. By combining standardized plant production, vector infection systems, transmission assays, and molecular diagnostics, these coordinated methods provide a unified framework for testing hypotheses about vector competence, pathogen–vector interactions, microbial co‐infections, host susceptibility, and environmental effects on transmission. Together, the procedures enable consistent and comparable assessment of C Las acquisition, dissemination, and transmission outcomes under controlled laboratory conditions. © 2026 The Author(s). Current Protocols published by Wiley Periodicals LLC. Basic Protocol 1 : Controlled laboratory method for assessing Candidatus Liberibacter asiaticus transmission by Diaphorina citri Support Protocol 1 : Generation of Candidatus Liberibacter asiaticus‐infected Citrus macrophylla and Diaphorina citri Basic Protocol 2 : Dellaporta total nucleic acid extraction Basic Protocol 3 : Citrus macrophylla propagation

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.

Abstract Deception Island fumaroles in Antarctica represent rare environments where extreme heat intersects with cryospheric and marine conditions, creating remarkable environmental gradients. From the near-boiling sediments, we reconstructed a high-quality metagenome-assembled genome (MAG) affiliated with the Pyrodictiaceae. Phylogenomic analyses revealed that this genome, proposed to represent Ca. Pyroantarcticum pellizari, forms a distinct lineage separated from known genera in the family. Functional annotation uncovered a versatile metabolic repertoire, including pathways for sulfur and nitrogen cycling, peptide and amino acid transport, and mixotrophic energy conservation. Stress-response systems such as reverse gyrase, thermosome, and small heat-shock proteins were complemented by lineage-specific genes related to membrane stability, metal detoxification, and Pyrodictiaceae-specific cannulae. These adaptations likely support survival under sharp temperature gradients, hydrogen sulfide emissions, and high metal concentrations at the volcanic–cryosphere–marine interface. Our findings expand the phylogenetic and ecological scope of Pyrodictiaceae, highlighting Antarctic marine volcanoes as unique refuges for hyperthermophiles and as valuable models for investigating life’s habitability under extreme temperatures.

Abstract Sulfoquinovose, a sulfonated sugar derived from the thylakoid membrane lipid sulfoquinovosyl diacylglycerol, is abundant in photosynthetic organisms and plays a key role in global sulfur cycling. Its degradation in nature is mediated by specialized bacteria, many of which rely on the enzyme sulfoquinovosidase (YihQ) to release sulfoquinovose from sulfoquinovosyl (diacyl)glycerol. Despite its ecological importance, the diversity and functional roles of sulfoquinovose-degrading microorganisms remain poorly characterized in natural environments. Here, we developed a yihQ-targeted amplicon sequencing approach to investigate the richness and distribution of SQ-degrading bacteria across selected environments. We revealed high richness of yihQ-containing microorganisms in the analyzed cow rumen samples, far exceeding that observed in human and mouse gut microbiomes, suggesting an important role of sulfoquinovose metabolism in ruminant digestion. Anoxic microcosm experiments with sulfoquinovose-amended rumen fluid revealed cooperative microbial degradation of sulfoquinovose to sulfide via isethionate cross-feeding. Amplicon sequencing and genome-resolved metagenomics and metatranscriptomics identified yet undescribed and uncultured sulfoquinovose-degrading taxa. Members of Caproiciproducens (Acutalibacteraceae), Candidatus Limivicinus (Oscillospiraceae), and Sphaerochaetaceae transcribed the isethionate-producing sulfo-transketolase pathway, whereas isethionate was likely respired by a Candidatus Mailhella bacterium (Desulfovibrionaceae). This study presents a functional gene-based assay for tracking environmental yihQ richness, highlights sulfoquinovose degradation as a central metabolic process in the cow rumen, describes previously unknown sulfoquinovose-metabolizing bacteria, and advances understanding of sulfur physiology in complex microbial communities.

Проведенные в 2019–2024 гг. исследования выявили в посевах сахарной свеклы в ЦЧР новое заболевание – фитоплазмоз сахарной свеклы (Candidatus Phytoplasma solani), проявляющееся на фоне засушливых погодных условий в конце вегетации. Изучение вирофорности цикадовых, питающихся на культуре, позволило сделать вывод, что переносчиками фитоплазмоза можно признать виды Pentrastridius leporinus, Hyalesthes obsoletus, Neoaliturus fenestratus. Их численность возрастает с увеличением среднесуточных и максимальных температур воздуха, суммы эффективных температур, при снижении осадков, ГТК и относительной влажности воздуха. Based on studies conducted in 20190–2024, a new disease was detected in sugar beet crops in the Central Black-Earth Region – phytoplasmosis of sugar beet (Candidatus Phytoplasma solani), which manifests itself against the background of dry weather conditions at the end of the growing season. The study of the virophoricity of cicadas feeding on culture led to the conclusion that the species Pentrastridius leporinus, Hyalesthes obsolete, and Neoaliturus fenestratus can be recognized as carriers of phytoplasmosis. Their number increases with an increase in average daily and maximum air temperatures, the sum of effective temperatures, with a decrease in precipitation, hydrothermal coefficient and relative humidity.