Cucumber green mottle mosaic virus (CGMMV) is a typical seed-borne tobamovirus that mainly infects cucurbit crops. Due to the rapid growth of international trade, CGMMV has spread worldwide and become a significant threat to cucurbit industry. Despite various studies focusing on the interaction between CGMMV and host plants, the molecular mechanism of CGMMV infection is still unclear. In this study, we utilized transcriptome and metabolome analyses to investigate the antiviral response of bottle gourd (Lagenaria siceraria) under CGMMV stress. The transcriptome analysis revealed that in comparison to mock-inoculated bottle gourd, 1929 differently expressed genes (DEGs) were identified in CGMMV-inoculated bottle gourd. Among them, 1397 genes were upregulated while 532 genes were downregulated. KEGG pathway enrichment indicated that the DEGs were mainly involved in pathways including the metabolic pathway, the biosynthesis of secondary metabolites, plant hormone signal transduction, plant–pathogen interaction, and starch and sucrose metabolism. The metabolome result showed that there were 76 differentially accumulated metabolites (DAMs), of which 69 metabolites were up-accumulated, and 7 metabolites were down-accumulated. These DAMs were clustered into several pathways, including biosynthesis of secondary metabolites, tyrosine metabolism, flavonoid biosynthesis, carbon metabolism, and plant hormone signal transduction. Combining the transcriptome and metabolome results, the genes and metabolites involved in the jasmonic acid and its derivatives (JAs) synthesis pathway were significantly induced upon CGMMV infection. The silencing of the allene oxide synthase (AOS) gene, which is the key gene involved in JAs synthesis, reduced CGMMV accumulation. These findings suggest that JAs may facilitate CGMMV infection in bottle gourd.

Cucumber green mottle mosaic virus (CGMMV, Tobamovirus viridimaculae) is a tobamovirus that induces leaf green mottling, mosaic patterns, bleaching, fruit sponginess, rotting, and malformation symptoms in various cucurbit crops. The underlying mechanisms by which CGMMV elicits these symptoms have yet to be elucidated. In the present study, we observed that the infection of CGMMV in bottle gourd, but not in N. benthamiana, led to the significant upregulation of a key gene involved in chlorophyll degradation, Chlorophyllase 1 (CLH1). This induction may be closely linked to chlorophyll degradation, particularly that of chlorophyll a (Clh a) in bottle gourd plants. Phylogenetic analysis showed that the amino acid sequence of BgCLH1 has a closer relationship with those of CLH1 from other cucurbit crops and has a relatively farther relationship with those of the well-studied CLH1 from Arabidopsis thaliana and Citrus sinensis. Further, confocal microscopy analysis indicated that BgCLH1 may be localized to the cytoplasm instead of the chloroplast. Moreover, silencing of the BgCLH1 gene not only reduced viral accumulation but also resulted in an increase in chlorophyll content. Similar results were also observed in watermelon, suggesting that this regulatory mechanism may be conserved across cucurbit crops. Our findings thus reveal a complex and intricate interplay between viral infection and the chlorophyll metabolic pathway.

Cucumber plants (Cucumis sativus) displaying symptoms of phyllody, sterility, and flower virescence were observed in fields located in the Dianbai district of Guangdong province, China. Total DNA was extracted from the symptomatic plants; this was followed by molecular detection using a set of universal phytoplasma primer pairs, R16mF2/mR1, P1/P7, and SecYF1(II)/SecYR1(II). This resulted in the PCR amplification of products corresponding to expected sizes of 1.4 kb, 1.8 kb and 1.7 kb, respectively. The 16S rDNA sequence obtained exhibited 100% similarity with the eggplant phyllody phytoplasma, the ‘Cleome rutidosperma’ witches’ broom phytoplasma, and the ‘Desmodium ovalifolium’ witches’ broom phytoplasma strain DeOWB, all of which belong to the 16SrII group. Phylogenetic analysis, based on the 16S rDNA gene and SecY gene sequences, confirmed the close affiliation of the detected phytoplasma isolate, tentatively designated as cucumber phyllody phytoplasma (CuPh) China isolate, with the 16SrII-A subgroup. Additionally, virtual restriction fragment length polymorphism (RFLP) analysis of the 16S rDNA sequence revealed a pattern that was identical to that of the 16SrII-A subgroup. This is the first report of cucumber phyllody phytoplasma in China.

Lettuce (Lactuca sativa L.) plants showing leaf curl and vein enation symptoms were found in Yunnan province, China. PCR detection with genus-specific primers revealed that symptomatic lettuce plants were infected with Begomovirus. The full-length viral component and satellite molecules were obtained by RCA, restriction enzyme digestion, PCR, cloning and DNA sequencing. A viral component (YN-2023-WJ) and three satellite molecules (YN-2023-WJ-alpha1, YN-2023-WJ-alpha2 and YN-2023-WJ-beta) were obtained from diseased lettuce plants. YN-2023-WJ exhibited the highest nt identity at 97.1% with pepper leaf curl Yunnan virus isolated from cigar plants. YN-2023-WJ-beta displayed the highest nt identity at 93.9% with tomato leaf curl China betasatellite. YN-2023-WJ-alpha1 showed the highest nt identity at 94.7% with ageratum yellow vein alphasatellite. YN-2023-WJ-alpha2 shared the highest nt identity at 75.6% with gossypium mustelinum symptomless alphasatellite and vernonia yellow vein Fujian alphasatellite. Based on the threshold for the classification of Begomovirus, Betasatellite and Alphasatellite, YN-2023-WJ was designated as a new isolate of PepLCYnV, YN-2023-WJ-beta as a new isolate of ToLCCNB and YN-2023-WJ-alpha1 as a new member of AYVA, whereas YN-2023-WJ-alpha2 was identified as a new geminialphasatellite species, for which the name pepper leaf curl Yunnan alphasatellite (PepLCYnA) is proposed. To the best of our knowledge, this is the first report of L. sativa L. infection by PepLCYnV associated with ToLCCNB, AYVA and PepLCYnA, and L. sativa L. is a new host plant of Begomovirus.

Desmodium styracifolium (Osb.) Merr., a member of the Leguminosae family, is an important medicinal plant widely used in traditional Chinese medicine. In September 2024, D. styracifolium plants exhibiting symptoms of little leaf and stunted growth were observed in a field of Zhanjiang, Guangdong province, China. Since the symptoms resembled those associated with phytoplasma infections, total DNA was extracted from the leaves of four symptomatic plants and one healthy plant for molecular identification. Universal primer pairs (P1/P7, R16mF2/mR1) for phytoplasma detection were used to amplify the 16S rDNA fragments (~1.8 kb and ~1.4 kb), while a specific primer pair secY-F/secY-R was employed to amplify a ~1.4 kb segment of the secY gene. Target fragments were successfully amplified from all symptomatic samples but not from the healthy control. These amplicons were cloned and sequenced. The obtained 16S rDNA sequence of D. styracifolium little leaf phytoplasma (DsLFP-GDZJ) showed the highest identity (99.67–100%) with strains of ‘Ca. Phytoplasma australasiaticum’ (subgroup 16SrII-A and 16SrII-D). Phylogenetic analysis also indicated that DsLFP-GDZJ formed a small evolutionary branch with strains of ‘Ca. Phytoplasma australasiaticum’ (subgroup 16SrII-A and 16SrII-D). Virtual RFLP (restriction fragment length polymorphism) analysis of the 16S rDNA sequence demonstrated DsLFP-GDZJ belongs to the 16SrII-A subgroup (GenBank accession number L33765). The secY gene sequence of DsLFP-GDZJ also showed the highest similarity and the closest relationship with those of the 16SrII-A subgroup phytoplasma strains. These results showed that DsLFP-GDZJ is a strain of ‘Ca. Phytoplasma australasiaticum’ (16SrII-A subgroup). To our knowledge, this is the first report of ‘Ca. Phytoplasma australasiaticum’—related phytoplasma associated with D. styracifolium little leaf disease in China, thereby establishing D. styracifolium (Osb.) Merr. as a new host plant of phytoplasma.