Oriental melon (Cucumis melo L. var. makuwa) is an important fruit for human consumption. However, this plant species is one of the most recalcitrant to genetic transformation. The lack of an efficient in vitro system limits the development of a reproducible genetic transformation protocol for Oriental melon. In this study, an efficient transgenic production method for Agrobacterium-mediated transformation using cotyledon explants of Oriental melon was developed. Cotyledon explants were pre-cultivated for two days in the dark, and the optimal conditions for transformation of melon were determined to be a bacteria concentration of OD600 0.6, inoculation for 30 min, and two days of co-cultivation. Transgenic melon plants were produced from kanamycin-resistant shoots. A total of 11 independent transgenic plants were regenerated with a transformation efficiency of 0.8% of the inoculated explants. The transgenic plants were phenotypically normal and fully fertile, which might be a consequence of the co-cultivation time.

Glossogyne tenuifolia Cassini (Hsiang-Ju in Chinese) is a perennial herb native to Taiwan. It was used in traditional Chinese medicine (TCM) as an antipyretic, anti-inflammatory, and hepatoprotective agent. Recent studies have shown that extracts of G. tenuifolia possess various bioactivities, including anti-oxidant, anti-inflammatory, immunomodulation, and anti-cancer properties. However, the pharmacological activities of G. tenuifolia essential oils have not been studied. In this study, we extracted essential oil from air-dried G. tenuifolia plants, then investigated the anti-inflammatory potential of G. tenuifolia essential oil (GTEO) on lipopolysaccharide (LPS)-induced inflammation in murine macrophage cells (RAW 264.7) in vitro. Treatment with GTEO (25, 50, and 100 μg/mL) significantly as well as dose-dependently inhibited LPS-induced pro-inflammatory molecules, such as nitric oxide (NO) and prostaglandin E₂ (PGE₂) production, without causing cytotoxicity. Q-PCR and immunoblotting analysis revealed that the inhibition of NO and PGE₂ was caused by downregulation of their corresponding mediator genes, inducible nitric oxide synthase (iNOS), and cyclooxygenase-2 (COX-2), respectively. Immunofluorescence and luciferase reporter assays revealed that the inhibition of iNOS and COX-2 genes by GTEO was associated with the suppression of nuclear export and transcriptional activation of the redox-sensitive transcription factor, nuclear factor -κB (NF-κB). In addition, GTEO treatment significantly inhibited phosphorylation and proteosomal degradation of the inhibitor of NF-κB (I-κBα), an endogenous repressor of NF-κB. Moreover, treatment with GTEO significantly blocked the LPS-mediated activation of inhibitory κB kinase α (IKKα), an upstream kinase of the I-κBα. Furthermore, p-cymene, β-myrcene, β-cedrene, cis-β-ocimene, α-pinene, and _D-limonene were represented as major components of GTEO. We found that treatment with p-cymene, α-pinene, and _D-limonene were significantly inhibiting LPS-induced NO production in RAW 264.7 cells. Taken together, these results strongly suggest that GTEO inhibits inflammation through the downregulation of NF-κB-mediated inflammatory genes and pro-inflammatory molecules in macrophage cells.

Alfalfa (Medicago sativa L.) is a perennial plant, which is high in nutritional value and resistant to environmental conditions, and it is one of most frequently preferred feed crop among the leguminous family. In this study, it was aimed to determine the genetic diversity of some alfalfa ecotypes and their varieties by DNA, protein, nucleus, and chromosome counts. The genetic distance between the populations of control (M. truncatula), five different cultivars (Alsancak, Bilensoy, Iside, Plato, Bilensoy82), and three different ecotypes (Erzurum, Muş, and Konya) was investigated by cytogenetic analysis, flow cytometry, simple sequence repeats (SSR), and SDS PAGE techniques. Cytogenetic analysis of these tested plants has verified the existence of expected levels such as diploid, triploid, and tetraploid as well as aneuploid (2n = 4x = 30) plants. Flow cytometry analysis have displayed that all of tested plants were tetraploid, whereas cytological analysis had either diploid, triploid, or tetraploid. Genetic diversity dendrogram was created using Erzurum, Muş, Konya, Bilensoy82, Alsancak, and Plato varieties. The Iside and Bilensoy were found to be morphogenetic in relationship. Our control plant, M. truncatula, did not have a similarity relationship with other ecotypes and cultivars. The total numbers of protein bands differed among tested plants from 140 kDA to 25 kDa. This paper first reports on the genetic variation of Turkish alfalfa plants by using detailed analysis techniques. This work provides important findings for the classification, conservation, and innovation of alfalfa germplasm resources.