The complete nucleotide sequence of hibiscus chlorotic ringspot virus (HCRSV) was determined. The genomic RNA (gRNA) is 3,911 nucleotides long and has the potential to encode seven viral proteins in the order of 28 (p28), 23 (p23), 81 (p81), 8 (p8), 9 (p9), 38 (p38), and 25 (p25) kDa. Excluding two unique open reading frames (ORFs) encoding p23 and p25, the ORFs encode proteins with high amino acid similarity to those of carmoviruses. In addition to gRNA, two 3′-coterminated subgenomic RNA (sgRNA) species were identified. Full-length cDNA clones derived from gRNA and sgRNA were constructed under the control of a T7 promoter. Both capped and uncapped transcripts derived from the full-length genomic cDNA clone were infectious. In vitro translation and mutagenesis assays confirmed that all the predicted ORFs except the ORF encoding p8 are translatable, and the two novel ORFs (those encoding p23 and p25) may be functionally indispensable for the viral infection cycle. Based on virion morphology and genome organization, we propose that HCRSV be classified as a new member of the genus Carmovirus in family Tombusviridae .

Wound-induced protein kinase (WIPK) is a tobacco (Nicotiana tabacum) mitogen-activated protein kinase known to play an essential role in defense against wounding and pathogens, although its downstream targets have yet to be clarified. This study identified a gene encoding a protein of 648 amino acids, which directly interacts with WIPK, designated as N. tabacum WIPK-interacting factor (NtWIF). The N-terminal region with approximately 250 amino acids showed a high similarity to the plant-specific DNA binding domain, B3, but no other similarity with known proteins. The C terminus of approximately 200 amino acids appeared to be essential for the interaction with WIPK, and a Luciferase-reporter gene assay using Bright Yellow 2 cells indicated the full-length protein to possess trans-activation activity, located to the middle region of approximately 200 amino acids. In vitro phosphorylation assays indicated that WIPK efficiently phosphorylates the full-length protein and the N terminus but not the C terminus. When full-length NtWIF was coexpressed with WIPK in Bright Yellow 2 cells, the Luciferase transcriptional activity increased up to 5-fold that of NtWIF alone, whereas no effect was observed with a kinase-deficient WIPK mutant. Transcripts of NtWIF began to simultaneously accumulate with those of WIPK 30 min after wounding and 1 h after the onset of hypersensitive response upon tobacco mosaic virus infection. These results suggest that NtWIF is a transcription factor that is directly phosphorylated by WIPK, thereby being activated for transcription of target gene(s) involved in wound and pathogen responses.

Silybin A and B are two major hepatoprotective flavonolignans produced predominantly in Silybum marianum fruits. Similar to other plant secondary metabolites, silybin production is enhanced by biotic or abiotic stresses under the expanse of plant growth and yield. In this study, a novel strategy for enhancing silybin production was investigated by applying a natural plant growth enhancer, moringa leaf extract (MLE), to the S. marianum plants cultivated under salinity abiotic stress. The experiment was conducted using a split-plot design with salinity as the main factor and MLE concentrations as the sub-factor. The individual and combined effects of salinity and MLE on the growth, yield, and silybin content of S. marianum were recorded. The MLE at concentrations of 5 g/L or 10 g/L was applied as foliar spray to S. marianum plants, which were irrigated with either tap water or saline (2000 ppm or 4000 ppm). Our results revealed that 10 g/L MLE effectively enhances the growth, yield, and silybin (A + B) content in S. marianum plants not exposed to salinity stress. On the other hand, for plants that were exposed to 4000 ppm salinity, the application of MLE was able to alleviate the salinity-induced adverse effects on some of the plant growth parameters but did not significantly increase their silybin (A + B) compositions. The current study also indicated that the increase in silybin contents was accompanied by the upregulation of the chalcone synthase 1 and 3 genes, which have been implicated in the synthesis of silybin.

ZnO nanoparticles (NPs) can be considered a highly efficient Zn source that has been widely used in agriculture to promote crop development and productivity. The turmeric (Curcuma longa) plant has several medical properties, and its rhizome is utilized as a spice in the food sector. In this work, C. longa leaves were sprayed with various concentrations of ZnO NPs to inspect their effect on growth, yield, and bioactive compound compositions. ZnO NPs considerably increased tumeric productivity, yield, and curcuminoid content versus the control treatment. The ZnO NPs concentration of 10 mgL⁻¹ was found to be the optimum concentration for producing the highest C. longa yields, while the concentration of 40 mgL⁻¹ promoted positive effects on photosynthetic pigments, bisdemethoxycurcumin, demethoxycurcumin, and curcumin contents. This demonstrates that ZnO nano-fertilizer promotes plant growth, yield characteristics, and curcuminoid component synthesis, and its application is therefore notably beneficial for progressive sustainable C. longa agriculture.