No abstract

Environmental factors such as low temperature (LT) and high humidity (HH) hinder plant growth and development in plastic tunnels and solar greenhouses in the cold season. In this study, we examined the effect of melatonin (MT) on shoot-based tolerance to LT and HH conditions in cucumber (Cucumis sativus) seedlings and explored its underlying mechanism. LT and HH stress inhibited growth and biomass accumulation, produced leaf chlorosis, led to oxidative stress, lowered chlorophyll and carotenoid contents, reduced photosynthetic and photosystem II (PSII) activities, and increased the level of intercellular carbon dioxide and the non-photochemical quenching of photosystem I (PSI) and PSII. However, foliar application of MT significantly improved the morphological indices and photosynthetic efficiency of cucumber seedlings, which entailed the elevation of electrolyte leakage, lipid peroxidation, and reactive oxygen species accumulation by boosting the antioxidant enzyme defense system under LT and HH conditions. Additionally, the measurement of nitrogen (N), magnesium (Mg), and iron (Fe) contents in roots and leaves showed that MT significantly augmented the nutrient uptake of cucumber seedlings exposed to LT and HH stresses. Furthermore, MT application increased the transcripts levels of genes encoding antioxidant enzymes under LT and HH conditions, whereas treatment with LT and HH suppressed these genes, suggesting that MT application increases the LT and HH tolerance of cucumber seedlings. Overall, our results suggest that MT application increases the tolerance of cucumber seedlings to LT and HH stress by enhancing the plant morphometric parameters, regulating PSI and PSII, and activating the antioxidant defense mechanism. Thus, the exogenous application of MT could be potentially employed as a strategy to improve the LT and HH tolerance of cucumber.

Papaya (Carica papaya L.) is an economically important tropical fruit tree with hermaphrodite, male and female sex types. Hermaphroditic plants are the major type used for papaya production because their fruits have more commercial advantages than those of female plants. Sex determination of the seedlings, or during the early growth stages, is very important for the papaya seedling industry. Thus far, the only method for determining the sex type of a papaya at the seedling stage has been DNA analysis. In this study, a molecular technique—based on DNA analysis—was developed for detecting male-hermaphrodite-specific markers to examine the papaya’s sex type. This method is based on the loop-mediated isothermal amplification (LAMP) and does not require prior DNA purification. The results show that the method is an easy, efficient, and inexpensive way to determine a papaya’s sex. This is the first report on the LAMP assay, using intact plant materials-without DNA purification-as samples for the analysis of sex determination of papaya. We found that using high-efficiency DNA polymerase was essential for successful DNA amplification, using trace intact plant material as a template DNA source.

Melanoma differentiation-associated gene 9 (MDA-9) is a small adaptor protein with tandem PDZ domains that promotes tumor progression and metastasis in various human cancers. However, it is difficult to develop drug-like small molecules with high affinity due to the narrow groove of the PDZ domains of MDA-9. Herein, we identified four novel hits targeting the PDZ1 and PDZ2 domains of MDA-9, namely PI1A, PI1B, PI2A, and PI2B, using a protein-observed nuclear magnetic resonance (NMR) fragment screening method. We also solved the crystal structure of the MDA-9 PDZ1 domain in complex with PI1B and characterized the binding poses of PDZ1-PI1A and PDZ2-PI2A, guided by transferred paramagnetic relaxation enhancement. The protein–ligand interaction modes were then cross-validated by the mutagenesis of the MDA-9 PDZ domains. Competitive fluorescence polarization experiments demonstrated that PI1A and PI2A blocked the binding of natural substrates to the PDZ1 and PDZ2 domains, respectively. Furthermore, these inhibitors exhibited low cellular toxicity, but suppressed the migration of MDA-MB-231 breast carcinoma cells, which recapitulated the phenotype of MDA-9 knockdown. Our work has paved the way for the development of potent inhibitors using structure-guided fragment ligation in the future.

No abstract