Pinofuranoxins A and B, Bioactive Trisubstituted Furanones Produced by the Invasive Pathogen Diplodia sapinea.

Two new bioactive trisubstituted furanones, named pinofuranoxins A and B (1 and 2), were isolated from Diplodia sapinea, a worldwide conifer pathogen causing severe disease. Pinofuranoxins A and B were characterized essentially by NMR and HRESIMS spectra, and their relative and absolute configurations were assigned by NOESY experiments and computational analyses of electronic circular dichroism spectra. They induced necrotic lesions on Hedera helix L., Phaseolus vulgaris L., and Quercus ilex L. Compound 1 completely inhibited the growth of Athelia rolfsii and Phytophthora cambivora, while 2 showed antioomycetes activity against P. cambivora. In the Artemia salina assay both toxins showed activity inducing larval mortality.

Isolation of a phytotoxic isocoumarin from Diaporthe eres-infected Hedera helix (English ivy) and synthesis of its phytotoxic analogs.

BACKGROUND: The fungus Diaporthe eres was isolated from a fungal pathogen-infected leaf of Hedera helix (English ivy) exhibiting necrosis. It is hypothesized that the causative fungus produces phytotoxins as evidenced by necrotic lesions on the leaves. RESULTS: The fungus was isolated and grown in Czapek Dox broth culture medium and potato dextrose broth culture medium and identified as Diaporthe eres. The ethyl acetate extracts of the culture broths were phytotoxic to lettuce (Lactuca sativa) and bentgrass (Agrostis stolonifera). 3,4-Dihydro-8-hydroxy-3,5-dimethylisocoumarin (1) and tyrosol (2) were isolated and identified as the phytotoxic constituents. Six analogs of 3,4-dihydro-isocoumarin were synthesized and shown to be phytotoxic. The synthesized 3,4-dihydro-8-hydroxy-3,7-dimethylisocoumarin and 3,4-dihydro-8-hydroxy-3,3,7-trimethylisocoumarin were two- to three-fold more phytotoxic than the naturally occurring 1 in a Lemna paucicostata growth bioassay. CONCLUSION: Synthesis and herbicidal activities of the several new analogs of 1 are reported for the first time. These promising molecules should be used as templates for synthesis and testing of more analogs. © 2017 Society of Chemical Industry.

Identification and validation of reference genes for quantitative real-time PCR studies in Hedera helix L.

Reference gene evaluation and selection are necessary steps in gene expression analysis, especially in new plant varieties, through reverse transcription quantitative real-time PCR (RT-qPCR). Hedera helix L. is an important traditional medicinal plant recorded in European Pharmacopoeia. Research on gene expression in H. helix has not been widely explored, and no RT-qPCR studies have been reported. Thus, it is important and necessary to identify and validate suitable reference genes to for normalizing RT-qPCR results. In our study, 14 candidate protein-coding reference genes were selected. Their expression stability in five tissues (root, stem, leaf, petiole and shoot tip) and under seven abiotic stress conditions (cold, heat, drought, salinity, UV-C irradiation, abscisic acid and methyl jasmonate) were evaluated using geNorm and NormFinder. This study is the first to evaluate the stability of reference genes in H. helix. The results show that different reference genes should be chosen for normalization on the basis of various experimental conditions. F-box was more stable than the other selected genes under all analysis conditions except ABA treatment; 40S was the most stable reference gene under ABA treatment; in contrast, EXP and UBQ were the most unstable reference genes. The expressions of HhSE and Hhß-AS, which are two genes related to the biosynthetic pathway of triterpenoid saponins, were also examined for reference genes in different tissues and under various cold stress conditions. The validation results confirmed the applicability and accuracy of reference genes. Additionally, this study provides a basis for the accurate and widespread use of RT-qPCR in selecting genes from the genome of H. helix.