Detection and identification of Bogia coconut syndrome phytoplasma from seed-associated tissues and seedlings of coconut (Cocos nucifera) and betel nut (Areca catechu).

Evidence for seed transmission of phytoplasmas has grown in several pathosystems including coconut (Cocos nucifera). Bogia coconut syndrome (BCS) is a disease associated with the lethal yellowing syndrome associated with the presence of 'Candidatus Phytoplasma noviguineense' that affects coconut, betel nut (Areca catechu) and bananas (Musa spp.) in Papua New Guinea. Coconut and betel nut drupes were sampled from BCS-infected areas in Papua New Guinea, dissected, the extracted nucleic acid was used in polymerase chain reaction (PCR), and loop mediated isothermal amplification (LAMP) used to check for presence of phytoplasma DNA. In a second study, drupes of both plant species were collected from multiple field sites and grown in insect-proof cages. Leaf samples taken at 6 months were also tested with PCR and LAMP. The studies of dissected coconut drupes detected phytoplasma DNA in several tissues including the embryo. Drupes from betel nut tested negative. Among the seedlings, evidence of possible seed transmission was found in both plant species. The results demonstrate the presence of 'Ca. P. noviguineense' in coconut drupes and seedlings, and in seedlings of betel nut; factors that need to be considered in ongoing management and containment efforts.

Complex multiple introductions drive fall armyworm invasions into Asia and Australia.

The fall armyworm (FAW) Spodoptera frugiperda is thought to have undergone a rapid 'west-to-east' spread since 2016 when it was first identified in western Africa. Between 2018 and 2020, it was recorded from South Asia (SA), Southeast Asia (SEA), East Asia (EA), and Pacific/Australia (PA). Population genomic analyses enabled the understanding of pathways, population sources, and gene flow in this notorious agricultural pest species. Using neutral single nucleotide polymorphic (SNP) DNA markers, we detected genome introgression that suggested most populations in this study were overwhelmingly C- and R-strain hybrids (n = 252/262). SNP and mitochondrial DNA markers identified multiple introductions that were most parsimoniously explained by anthropogenic-assisted spread, i.e., associated with international trade of live/fresh plants and plant products, and involved 'bridgehead populations' in countries to enable successful pest establishment in neighbouring countries. Distinct population genomic signatures between Myanmar and China do not support the 'African origin spread' nor the 'Myanmar source population to China' hypotheses. Significant genetic differentiation between populations from different Australian states supported multiple pathways involving distinct SEA populations. Our study identified Asia as a biosecurity hotspot and a FAW genetic melting pot, and demonstrated the use of genome analysis to disentangle preventable human-assisted pest introductions from unpreventable natural pest spread.

Planthopper Transmission of Ramu Stunt Virus, a Tenuivirus Causing the Sugarcane Disease Ramu Stunt, and its Distribution in Papua New Guinea.

Ramu stunt is a serious disease of sugarcane, currently only reported from Papua New Guinea. It is found in both commercial sugarcane grown on the Ramu Agri Industries Limited (RAIL) estate and in chewing canes (Saccharum officinarum L.) grown in village gardens. The vector of Ramu stunt disease is the island sugarcane planthopper, Eumetopina flavipes Muir. Here we report on the successful transmission of Ramu stunt using E. flavipes and verify that the disease is caused by Ramu stunt virus, a virus with homology to the genus Tenuivirus. Diagnostic reverse transcription PCR screening, with partial genome sequencing and viral protein characterization, was used for confirmation. Disease surveys were undertaken on the RAIL estate, along roadsides, and in village gardens in parts of Papua New Guinea. When the disease was identified, partial genome sequencing of the virus was performed to assess the extent of genome variability among isolates. The disease was less common than predicted from early surveys based on symptoms alone, and genotypic variation was associated with geographic location.

Detection by next generation sequencing of a multi-segmented viral genome from sugarcane associated with Ramu stunt disease.

Ramu stunt disease of sugarcane was first reported in Papua New Guinea in the mid 1980s. The disease can reduce sugarcane yields significantly and causes severe stunting and mortality in highly susceptible cultivars. The causal agent of Ramu stunt has been investigated but its characterization has not been completed. Sugarcane cv. Ragnar from Papua New Guinea with symptoms of Ramu stunt was analyzed by next generation sequencing. Total RNA was extracted and whole transcriptome shotgun sequencing was performed using an Illumina platform. Over thirty-seven million reads with an average length of 100 nucleotides were obtained. More than eighteen thousand contigs were assembled and subjected to BLASTX analysis. Twenty-one contigs were virus related and six were associated with plant viruses. The BLAST algorithms revealed sequence similarity to Tenuivirus and Phlebovirus, genera of viruses whose members contain genomes consisting of multiple RNA segments. The six contigs derived from the RNA sequencing data correspond to six RNAs that compose the Ramu stunt virus genome. Primers were designed for each of the six RNAs and RT-PCR amplicons were obtained only from the symptomatic sugarcane. There was concordance between the sequence data of the contigs obtained from the NGS and that of the amplicons obtained by RT-PCR. The NGS approach allowed us to determine the complete genomic sequence of Ramu stunt virus. It is likely that this virus is the causal agent of Ramu stunt disease.