Changes of phenolic compounds in LebZIP2-overexpressing transgenic plants

The bZIP gene is a transcription factor that plays various roles in relation to plant stress and hormone signaling. This gene is also involved in plant environmental stress and herbicide tolerance. We generated Nicotiana benthamiana transgenic plants with LebZIP2-encoding gene isolated from tomatoes using Agrobacterium-mediated transformation. Transgenic seeds harvested from these T0 transgenic plants were grown and examined for gene transfer and changes in phenolic compounds in the T1 generation. RT-PCR analysis using a primer specific to the LebZIP gene confirmed that the gene was transferred to the T1 generation. We analyzed the increase and decrease tendency for 30 phenolic compounds using the T1 generation-transgenic plants and investigated the mechanism between the specifically increased compound and LebZIP2 gene. Gallic acid, homogentisic acid, protocatechuic acid, myricetin, t-cinnamic acid, and b-resorcyclic acid were identified as the phenolic compounds that increased in T1 transgenic plants overexpressing the LebZIP gene. Among these, homogentisic acid at 246.75-1055.19 µg/g, was increased by 2-5 fold in the T1 transgenic plants compared to the control. Protocatechuic acid was found at 1640.54-2456.00 µg/g and was increased by 2-4 fold in T1 transgenic plants. t-Cinnamic acid was present in a small amount of 23.14 µg/g in the control, whereas it was 102.19-135.47 µg/g in T1 transgenic plants, showing an increase of 4-5 folds. These results indicated that homogentisic acid, protocatechuic acid, and t-cinnamic acid among the 30 phenolic compounds analyzed, were significantly increased in LebZIP2-overexpressing T1 transgenic plants, and support the evidence that the LebZIP2 gene is significantly involved in the increment of three phenolic compounds.

Elicitation Enhanced the Production of Phenolic Compounds and Biological Activities in Hairy Root Cultures of Bitter melon ( Momordica charantia L.)

ABSTRACT Momordica charantia (Cucurbitaceae) is an important vegetable and also medicinal crop which produces the bioactive compounds for various biological activities with potential uses in human health. The present investigation relates to elicitors of jasmonic acid (JA) and salicylic acid (SA) to enhance biomass accumulation and phenolic compound production in hairy root cultures of M. charantia. Hairy root cultures were elicited with JA and SA at 0, 25, 50 and 100 μM concentrations respectively. The adding of elicitation to the hairy root cultures on the 15th day of culture and the roots were harvested on day 25. Cultures supplemented with 100 μM JA and SA enhanced the phenolic compounds significantly compared to that of non-elicited hairy root cultures. The biomass of hairy root culture significantly increased by SA whereas decreased in JA elicitation at 100 μM. JA and SA-elicited hairy root cultures significantly produced a higher amount of phenolic compounds (12811.23 and 11939.37µg/g), total phenolic (4.1 and 3.7 mg/g) and flavonoid (3.5 and 3.2 mg/g) contents than non-elicited hairy root cultures (10964.25 µg/g, 2.8 and 2.5 mg/g). JA and SA-elicited hairy root cultures were significantly higher antioxidant activity of DPPH (84 and 78%), reducing potential (0.53 and 0.48), phosphomolybdenum (3.6 and 3.2 mg/g) and ferrous ion chelating assays (80 and 74%) than non-elicited hairy root cultures. The higher antimicrobial and anticancer activity were exhibited in JA and SA-elicited than non-elicited hairy root cultures. This protocol can be developed for the production of phenolic compounds from JA and SA-elicited hairy root cultures.

Phenolic compound production and biological activities from in vitro regenerated plants of gherkin (Cucumis anguria L. )

Background The effect of polyamines (PAs) along with cytokinins (TDZ and BAP) and auxin (IBA) was induced by the multiple shoot regeneration from leaf explants of gherkin (Cucumis anguria L.). The polyphenolic content, antioxidant and antibacterial potential were studied from in vitro regenerated and in vivo plants. Results Murashige and Skoog (MS) medium supplemented with 3% sucrose containing a combination of 3.0 µM TDZ, 1.0 µM IBA and 75 µM spermidine induced maximum number of shoots (45 shoots per explant) was achieved. Regenerated shoots elongated in shoot elongation medium containing 1.5 µM GA3 and 50 µM spermine. The well-developed shoots were transferred to root induction medium containing 1.0 µM IBA and 50 µM putrescine. Rooted plants were hardened and successfully established in soil with a 95% survival rate. Twenty-five phenolic compounds were identified by ultra-performance liquid chromatography (UPLC) analysis The individual polyphenolic compounds, total phenolic and flavonoid contents, antioxidant and antibacterial potential were significantly higher with in vitro regenerated plants than in vivo plants. Conclusions Plant growth regulators (PGRs) and PAs had a significant effect on in vitro plant regeneration and also a biochemical accumulation of flavonols, hydroxybenzoic and hydroxycinnamic acid derivatives in C. anguria. Due to these metabolic variations, the antioxidant and antibacterial activities were increased with in vitro regenerated plants than in vivo plants. This is the first report describing the production of phenolic compounds and biological activities from in vitro and in vivo regenerated plants of C. anguria.

The effect of nitrogen and potassium fertilizers on growth parameters and carbohydrate contents of sweet sorghum cultivars.

Sweet sorghum is tolerant to high temperature and drought and can be considered as an alternative crop to sugar beet and maize in Iran. In this study, the effects of nitrogen and potassium fertilizers on growth parameters including stem height, stem diameter, stem fresh weight, total fresh weight; carbohydrate contents including total sugar, brix value, sucrose content and purify; and juice extract of two sweet sorghum cultivars were determined. Three rates of N-fertilizer (0, 90, 180 kg urea ha(-1)) and two rates of K fertilizer (0 and 50 kg potassium sulfate ha(-1)) assigned as main plots and two sweet sorghum cultivars (Rio and Keller) as subplots. Growth parameters at soft dough and physiological maturity stages and carbohydrate contents at physiological maturity stage were determined. Results showed that application of 180 kg urea ha(-1) as compared to control at physiological maturity significantly (p < 0.01) increased stem height (12.65%), stem fresh weight (24.57%), total fresh weight (78.22%), total sugar (39.25%), sucrose content (9%) and juice extract (34.96%). Application of 50 kg potassium sulfate ha(-1) increased (p < 0.05) stem fresh weight (24.33%), total fresh weight (25.44%), total sugar (10.50%), and juice extract (9%) at physiological maturity. The highest growth parameters, carbohydrate contents and juice extract were obtained with the application of 180 kg urea ha(-1) and 50 kg potassium sulfate ha(-1) using cultivar (cv) Keller. The best results were taken with the application of both fertilizers.

Pleurotus sajor-caju HSP100 complements a thermotolerance defect in hsp104 mutant Saccharomyces cerevisiae.

A putative Hsp100 gene was cloned from the fungus Pleurotus sajor-caju. mRNA expression studies demonstrated that this gene (designated PsHsp100) is highly induced by high temperature,induced less strongly by exposure to ethanol, and not induced by drought or salinity. Heat shock induction is detectable at 37 degrees C and reaches a maximum level at 42 degrees C. PsHsp100 mRNA levels sharply increased within 15 min of exposure to high temperature, and reached a maximum expression level at 2 h that was maintained for several hours. These results indicate that PsHsp100 could work at an early step in thermotolerance. To examine its function, PsHsp100 was transformed into a temperature-sensitive hsp104 deletion mutant Saccharomycetes cerivisiae strain to test the hypothesis that PsHSP100 is an protein that functions in thermotolerance. Overexpression of PsHSP100 complemented the thermotolerance defect of the hsp104 mutant yeast, allowing them being survive even at 50 degree C for 4 h. These results indicate that PsHSP100 protein is functional as an HSP100 in yeast and could play and important role in thermotolerance in P. sajor-caju.