Phytotoxic Effects of Catnip (Nepeta meyeri Benth.) on Early Growth Stages Development and Infection Potential of Field Dodder (Cuscuta campestris Yunck).

Dodder (Cuscuta campestris Yunck.) is one of the most devastating parasitic plants, which reduces quantity and quality of crops. The inhibitory effect of catnip (Nepeta meyeri Benth.) extracts on germination and some seedling characteristics of the C. campestris were investigated in three phases in a laboratory and greenhouse. Aqueous extracts from different organs of N. meyeri were used in bioassays. The N. meyeri extracts reduced germination percent, root and shoot growth, and dry weight of C. campestris seedlings. Moreover, results showed an inhibitory effect of the N. meyeri extracts on the activity of alpha-amylase, protease, and beta-1,3-glucanase enzymes in C. campestris germinating seeds. Under greenhouse conditions, C. campestris seeds were planted with 30-day-old alfalfa plants and irrigated with N. meyeri extracts. The application of extracts from different organs of N. meyeri reduced emergence percent and length of stem and hampered C. campestris attachment to alfalfa. N. meyeri extracts also inhibited the activity of antioxidant enzymes and increased the accumulation of hydrogen peroxide and the malondialdehyde in C. campestris seedlings. The strongest inhibitory effects were observed from flower, leaf, and stem extracts of N. meyeri, respectively. However, after C. campestris attachment to alfalfa plants, treatment by N. meyeri extracts did not exhibit any effect on infestation efficiency and C. campestris growth traits. According to these findings, N. meyeri extract, especially from flower and leaf, may be recommended as a potent bio-control agent to control germination and early stage development of C. campestris.

Physiological and Molecular Aspects of Two Thymus Species Differently Sensitive to Drought Stress.

Drought is one of the most important threats to plants and agriculture. Here, the effects of four drought levels (90%, 55%, 40%, and 25% field capacity) on the relative water content (RWC), chlorophyll and carotenoids levels, and mRNA gene expression of metabolic enzymes in Thymus vulgaris (as sensitive to drought) and Thymus kotschyanus (as a drought-tolerant species) were evaluated. The physiological results showed that the treatment predominantly affected the RWC, chlorophyll, and carotenoids content. The gene expression analysis demonstrated that moderate and severe drought stress had greater effects on the expression of histone deacetylase-6 (HDA-6) and acetyl-CoA synthetase in both Thymus species. Pyruvate decarboxylase-1 (PDC-1) was upregulated in Thymus vulgaris at high drought levels. Finally, succinyl CoA ligase was not affected by drought stress in either species. Data confirmed water stress is able to alter the gene expression of specific enzymes. Furthermore, our results suggest that PDC-1 expression is independent from HDA-6 and the increased expression of ACS can be due to the activation of new pathways involved in carbohydrate production.

Combined effect of putrescine and mycorrhizal fungi in phytoremediation of Lallemantia iberica in Pb-contaminated soils.

As soil contamination with heavy metals is increasing and polyamines have roles in the growth of mycorrhiza and plants, it is important to study phytoremediation, growth, tolerance, and mycorrhization in Lallemantia iberica as a multi-purpose plant, by the application of putrescine along with mycorrhiza in Pb-contaminated soils. For this purpose, the study was performed in a factorial arrangement with Pb (0, 300, 600, and 900 mg Pb/kg soil), mycorrhiza (non-inoculation, Funneliformis mosseae (Fm), and Rhizophagus intraradices (Ri)), and putrescine (0, 0.5, and 1 mM) in a greenhouse. Results showed that antioxidant activities, plant Pb, and mycorrhizal features enhanced, while transfer factor (TF), biomass, and tolerance decreased under Pb levels. Mycorrhiza improved growth, greenness, defense, and tolerance and reduced TF, Pb, and H2O2 content under Pb stress. Putrescine (0.5 mM) increased catalase activity, biomass, and colonization and reduced Pb content and TF under Pb levels. Combination of 0.5 mM putrescine with Fm increased shoot biomass (13%), peroxidase (17.2%), root P (7.5%), shoot tolerance (14.4%), colonization (5.1%), and hyphal width (5.5%) and decreased malondialdehyde (20.5%) and shoot Pb content (28.1%). Putrescine (1 mM) had negative effects on all traits in combination with Ri but not with Fm. Combination of putrescine and Fm showed more efficiency in decreasing Pb content in L. iberica and was effective in phytostabilization. It is generally concluded that 0.5 mM putrescine was the beneficial concentration in combination with mycorrhiza, Pb stress, and single use to improve plant performance, and Fm was a useful species for improving the growth and tolerance of L. iberica under Pb levels.

Effect of drought stress on metabolite adjustments in drought tolerant and sensitive thyme.

Drought is one of the most important threats to plants and agriculture; therefore, understanding of the mechanism of drought tolerance is crucial for breeding of drought tolerant plants. Here, we assessed effects of four levels of drought (90%, 55%, 40% and 25% FC) on some physiological criteria and metabolite adjustment of two different drought-responsive thyme plants (Thymus vulgaris as drought sensitive and T. Kotschyanus as drought tolerant species), using 1H-NMR. Among three physiological parameters and 18 identified metabolites, species × treatment effects were significant (P ≤ 0.01) for leaf temperature, acetic acid, citric acid, fumaric acid, malic acid, succinic acid, fructose, sucrose and serine. RWC, chlorophyll and carotenoids content, glucose, alanine and choline were affected by simple effects of species and treatment. Correlation analysis revealed that there is a different correlation between physiological parameters and metabolites in both species. This analysis also revealed that, by ignoring the correlation between malic acid and succinic acid in T. vulgaris, there was no significant correlation between TCA intermediate in both species. According to results, sugars, amino acid and energy metabolism were affected by drought and, among them, TCA intermediates had more alternation in two studied species so, this cycle and its intermediates probably have more prominent role than other identified metabolites in the induction of drought tolerance.

Evaluation and validation of housekeeping genes in two contrast species of thyme plant to drought stress using real-time PCR.

To decrease errors and increase accuracy and reliability of quantitative real-time PCR (qRT-PCR) results, the use of a reference gene is inevitable. Despite the industrial importance of genus Thymus, not any validated reference gene has not been reported for T. kotschyanus and T. vulgaris which could limit such investigations. In this study, the expression stability of seven housekeeping genes including Actin, Cyclophilin-18, elongation factor-1A, glyceraldehyde-3-phosphate dehydrogenase, 18S ribosomal RNA, Cullin, and Polypyrimidine tract-binding protein were evaluated in T. kotschyanus and T. vulgaris which grown at four levels of drought stress using geNorm, NormFinder, and BestKeeper algorithms. Histone deacetylase-6 (HDA-6) gene was also used for validation of evaluated reference genes. In T. vulgaris, all of the algorithms similarly ranked elongation factor-1A and glyceraldehyde-3-phosphate dehydrogenase as the two most stably expressed genes. In T. kotschyanus, only NormFinder and BestKeeper had a similar ranking and identified Actin and glyceraldehyde-3-phosphate dehydrogenase as the two most stably expressed genes, but geNorm algorithm ranked elongation factor-1A and glyceraldehyde-3-phosphate dehydrogenase as the best two reference genes. On the other hand, all algorithms ranked 18S rRNA and Cyclophilin-18 as the least stable genes in T. kotschyanus and T. vulgaris, respectively. Validation results indicated that there was a significant change (0.53-3.19 fold change) in relative expression of HDA-6 normalized by the best stable gene compare to the least ranked gene. Our study presented the first systematic validation of reference gene(s) selection in T. kotschyanus and T. vulgaris and provided useful information to obtain more accurate qRT-PCR results in these species.