Areca palm velarivirus 1 infection caused disassembly of chloroplast and reduction of photosynthesis in areca palm.

The expansion of betel palm cultivation is driven by rising demand for betel nut, yet this growth is accompanied by challenges such as decreased agricultural biodiversity and the spread of infectious pathogens. Among these, Yellow Leaf Disease (YLD) emerges as a prominent threat to betel palm plantation. Areca Palm Velarivirus 1 (APV1) has been identified as a primary causative agent of YLD, precipitating leaf yellowing, stunted growth, and diminished yield. However, the precise mechanisms underlying APV1-induced damage remain elusive. Our study elucidates that APV1 infiltrates chloroplasts, instigating severe damage and consequential reductions in chlorophyll a/b and carotene levels, alongside notable declines in photosynthetic efficiency. Moreover, APV1 infection exerts broad regulatory effects on gene expression, particularly suppressing key genes implicated in chloroplast function and photosynthesis. These disruptions correlate with growth retardation, yield diminishment, and compromised nut quality. Intriguingly, the paradoxical destruction of the host's photosynthetic machinery by APV1 prompts inquiry into its evolutionary rationale, given the virus's dependence on host resources for replication and proliferation. Our findings reveal that APV1-induced leaf yellowing acts as a beacon for transmission vectors, hinting at a nuanced "host-pathogen-vector co-evolutionary" dynamic.

Identification and characterization of two closely related virga-like viruses latently infecting rubber trees (Hevea brasiliensis).

A novel virga-like virus, provisionally named Rubber tree latent virus 2 (RTLV2), was identified from rubber tree (Hevea brasiliensis). It is a close relative of the previously reported Rubber tree latent virus 1 (RTLV1). The complete genomes of RTLV1 and RTLV2 were sequenced and comparatively analyzed in terms of genome organization, putative gene products and phylogenetic relationship. Both RTLV1 and RTLV2 have positive-sense single-stranded RNA genomes that encode seven open reading frames (ORFs), forming a similar genomic layout. In phylogenetic analyses based on replicase and coat protein amino acid sequences, RTLV1 and RTLV2 were clustered with unclassified virga-like viruses. They are distinct from currently recognized plant virus families. RTLV1 and RTLV2 can be distinguished from members of Virgaviridae by the presence of a putative coat protein duplex and a poly(A) tail at the 3'-terminus. The authenticity of RTLV1 and RTLV2 as infectious viruses was confirmed through field investigations and transmissibility assays. In conclusion, RTLV1 and RTLV2 represent a novel plant virus group that does not readily fit into current virus families.

Identification and molecular characterization of a novel member of the genus Totivirus from Areca catechu L.

In previous work, RNA-seq was applied to identify the causal agent of yellow leaf disease (YLD) in areca palm (Areca catechu L.), resulting in the identification of areca palm velarivirus 1 (APV1) associated with YLD. Additionally, RNA-seq revealed a totivirus-like virus in areca palm. This work revealed that the totivirus-like virus is prevalent in asymptomatic areca palms. Therefore, it was tentatively named "areca palm latent totivirus 1" (APLTV1). The complete genome sequence of APLTV1 was determined and found to be 4754 base pairs (bp) in length, containing two ORFs whose encoded proteins share 55% and 69% amino acid (aa) sequence identity with the capsid protein (CP) and RNA-dependent RNA polymerase (RdRp), respectively, of Bursera graveolens-associated totivirus 1 (BgAT1). Phylogenetic analysis based on alignment of the CP and RdRp sequences revealed that APLTV1 clustered with other members of the genus Totivirus, suggesting that APLTV1 represents a novel species of the genus Totivirus, family Totiviridae.

Effect of temperature on yellow leaf disease symptoms and its associated areca palm velarivirus 1 titer in areca palm (Areca catechu L.).

Yellow leaf disease (YLD) has been a major limiting factor threatening areca palm commonly known as betel palm (Areca catechu L.) plantations in Hainan, China. The YLD disease is closely associated with areca palm velarivirus 1 (APV1), which belongs to the family Closteroviridae. YLD-affected betel palms show more serious yellowing symptoms in winter than in summer based on anecdotal observations. In the present work, the underlying mechanism was investigated. We first observed that the severity of YLD symptoms was closely related with the APV1 viral titer determined by qRT-PCR and ELISA under natural conditions. To further investigate whether temperature plays a key role in APV1 accumulation, the areca palm seedlings were artificially inoculated with APV1-positive mealybugs (Ferrisia virgata) and then cultivated under controlled conditions. According to our results, the YLD symptoms severity in inoculated seedlings were closely associated with temperature, e.g., severest symptoms at low temperature (16/22 ± 2°C, night/day), severer symptoms at room temperature (24/26 ± 2°C, night/day), while moderate symptoms at high temperature (27/34 ± 2°C, night/day). The qRT-PCR and ELISA results showed that APV1 titer accumulates significantly abundant at low temperature as compared to high and room temperatures. In conclusion, this is the first report about the temperature effects on the symptoms severity of YLD and APV1 titer, which may have important implications for the epidemiology of YLD.

Transmission of Areca Palm Velarivirus 1 by Mealybugs Causes Yellow Leaf Disease in Betel Palm (Areca catechu).

Yellow leaf disease (YLD) is the most destructive disease of betel palm (Areca catechu). A strong association between YLD and areca palm velarivirus 1 (APV1) has been observed. However, the causal relationship between APV1 and disease, and the transmission mode, warrant further investigation. This work showed that APV1 was transmitted by both Ferrisia virgata and Pseudococcus cryptus mealybugs and caused YLD symptoms in betel palm seedlings; therefore, we demonstrate that APV1 is a causal agent of YLD. APV1 was detected in the stylets, foreguts, midguts, and hindguts of the vectors via both immunocapture reverse transcription PCR and immunofluorescence assays. APV1 was not transmitted transovarially from viruliferous female F. virgata to their progeny. In summary, the transmission of APV1 by F. virgata may occur in a noncirculative, semipersistent manner. This study fills important gaps in our knowledge of velarivirus transmission, which is critical for developing YLD management practices.

Genomic diversity of Areca Palm Velarivirus 1 (APV1) in Areca palm (Areca catechu) plantations in Hainan, China.

BACKGROUND: Areca palm (Areca catechu L.) is an important commercial crop in southeast Asia, but its cultivation is threatened by yellowing leaf disease (YLD). Areca palm velarivirus 1 (APV1) was recently associated with YLD, but little is known regarding its population and genetic diversity. To assess the diversity of YLD, the APV1 genome was sequenced in YLD samples collected from different sites in Hainan. RESULTS: Twenty new and complete APV1 genomes were identified. The APV1 isolates had highly conserved sequences in seven open reading frames (ORFs; > 95% nucleotide [nt] identity) at the 3' terminal, but there was diversity (81-87% nt identity) in three ORFs at the 5' terminal. Phylogenetic analysis divided the APV1 isolates into three phylogroups, with 16 isolates (> 70%) in phylogroup A. Mixed infections with different genotypes in the same tree were identified; this was closely correlated with higher levels of genetic recombination. CONCLUSIONS: Phylogroup A is the most prevalent APV1 genotype in areca palm plantations in Hainan, China. Mixed infection with different genotypes can lead to genomic recombination of APV1. Our data provide a foundation for accurate diagnostics, characterization of etiology, and elucidation of the evolutionary relationships of APV1 populations.

Prevalence of Yellow Leaf Disease (YLD) and its Associated Areca Palm Velarivirus 1 (APV1) in Betel Palm (Areca catechu) Plantations in Hainan, China.

Yellow leaf disease (YLD) is an economically important disease affecting betel palm in several countries, the cause of which remains unclear despite associations with putative agents, including phytoplasmas. In this study, we screened the potential casual agents associated with YLD in Hainan, China using next-generation sequencing and revealed the association of areca palm velarivirus 1 (APV1) with the YLD-affected palm. The complete genome of the APV1-WNY isolate was determined to be 17,546 nucleotides in length, approximately 1.5 kb longer than the previously reported APV1_HN genome. Transmission electron microscopy showed that APV1 particles are flexuous and filamentous, a typical morphology of species in the Closteroviridae family. Comparison of symptomatic and symptomless tree populations showed a strong association between APV1 and YLD. APV1 was detected in Pseudococcus sp. mealybugs sampled from YLD-affected trees in many locations, suggesting that mealybugs are a potential transmission vector for APV1. Although further studies are needed to confirm a causal relationship, these results provide timely information for the prevention and management of YLD associated with APV1.

Complete genome sequence of a novel capillovirus infecting Hevea brasiliensis in China.

Tapping panel dryness (TPD) is a complex disorder that causes partial or complete cessation of latex drainage upon tapping of rubber trees (Hevea brasiliensis). In this work, we determined the complete genome sequences of a novel virus identified in a rubber tree with TPD syndrome in China. The genome of the virus consists of 6811 nt and possesses two overlapping open reading frames (ORF1 and ORF2), encoding a polyprotein and a movement protein, respectively. The polyprotein shares 37% amino acid sequence identity with cherry virus A (CVA, ARQ83874.1) over 99% coverage. The genome architecture is similar to that of members of the genus Capillovirus (family Betaflexiviridae). Phylogenetic analysis of the replicase proteins showed that the virus clustered together with members of the genus Capillovirus. The new virus is tentatively called "rubber tree virus 1" (RTV1). RTV1 is the first virus reported to infect rubber trees. This work lays a foundation for research into finding the potential causal agent of TPD in Hevea brasiliensis.