Proteome Changes in Stem Tissues of Sunflower Lines Inoculated with Culture Filtrate of Sclerotinia sclerotiorum.

BACKGROUND: Sclerotinia sclerotiorum (Lib.) de Bary cause a deleterious disease on sunflower plants. Oxalic acid is the main pathogenicity factor of S. sclerotiorum. Two dimensional gel electrophoresis and mass spectrometry have been used in several studies to investigate molecular changes that occur in the plants in response to S. sclerotiorum infection. Comparing responses of resistant and susceptible lines upon pathogen infection provided novel information regarding defense mechanisms against this necrotrophic pathogen. OBJECTIVES: The present study reports proteome changes of partially resistant and susceptible sunflower lines under pathogen's culture filtrate treatment, resulting in the characterization of up- and down- regulated proteins. MATERIAL AND METHODS: Sunflower partially resistant and susceptible lines with two true leaves were exposed to fungus culture filtrate. The stems of treated and untreated plants were sampled at 24, 48 and 72 hours after treatment for two-dimensional electrophoresis. Twenty spots showed more than 1.5-fold change in abundance were subjected to MALDI/TOF-TOF MS for further analysis. RESULTS: The identified proteins were categorized into several classes including carbohydrate and energy metabolism (25%), cellular metabolic process (15%), stress response (15%), plant cell wall biogenesis (10%), photosynthesis (10%), protein metabolism (10%), unknown function (10%) and redox homeostasis (5%). CONCLUSIONS: Our proteomic investigation demonstrates an increase in the expression of proteins only in partially resistant line, such as proteins involved in carbohydrate metabolism and plant defense responses (malate dehydrogenase and peroxidase), metabolic process (adenosine kinase), regulating cell redox homeostasis (disulfide isomerase) and lignin biosynthetic process (laccase). Moreover, the expression of pyrroline-5-carboxylate reductase, involved in proline biosynthesis, was significantly changed in both sunflower lines in response to pathogen culture filtrate. Proteins which were only up-regulated in the partially resistant lines might have a significant role in mediating the defense against Sclerotinia and could be considered for enhancing resistance against this devastating pathogen.

Bioactive Polyphenols Modulate Enzymes Involved in Grapevine Pathogenesis and Chitinase Activity at Increasing Complexity Levels.

The reduction of synthetic chemistry use in modern viticulture relies on either the biological control of microorganisms or the induction of pathogenesis-related proteins. In the present study, the effects of hydro-alcoholic plant extracts (PEs) (i.e., by-products of Vitis vinifera L., leaves of Olea europaea L. and Ailanthus altissima (Mill.) Swingle) were tested on purified enzymes activity involved in plant-pathogen interactions. The polyphenolic composition was assayed and analyzed to characterize the extract profiles. In addition, suspension cell cultures of grapevine were treated with PEs to study their modulation of chitinase activity. Application of grape marc's PE enhanced chitinase activity at 4 g L-1. Additionally, foliar treatment of grape marc's PE at two doses (4 g L-1 and 800 g L-1) on grapevine cuttings induced a concentration-dependent stimulation of chitinase activity. The obtained results showed that the application of bioactive compounds based on PEs, rich in phenolic compounds, was effective both at in vitro and ex/in vivo level. The overall effects of PEs on plant-pathogen interaction were further discussed by applying a multi-criteria decision analysis, showing that grape marc was the most effective extract.

The Effect of Oxalic Acid, the Pathogenicity Factor of Sclerotinia Sclerotiorum on the Two Susceptible and Moderately Resistant Lines of Sunflower.

BACKGROUND: One of the main sunflower diseases is the white mold Sclerotinia sclerotiorum. The oxalic acid (OA), which is one of the main pathogenicity factors of this fungus, beside the direct toxicity on the host, has other functions such as the disruption of the cell wall and chelating out the calcium ions. OBJECTIVES: Regarding the importance of this disease, it is important to study the reactions of the plant against OA which is a nonspecific toxin of many other necrotrophic fungi. MATERIALS AND METHODS: In this study, two susceptible and moderately resistant sunflower lines were inoculated with OA and samples at the first leaf stage were collected within the intervals of 2, 6, 12 and 24 hours post inoculation. The expression of five genes related to tricarboxylic acid cycle, including citrate synthase, fumarase, iso-citrate lyase, malate synthase and malate dehydrogenase was studied under OA treatment. RESULTS: Two hours after the inoculation, no significant change was observed in the expression of the five studied genes in the moderately resistant line. The iso-citrate lyase gene, which is related to glyoxylate cycle (a variation of the tricarboxylic acid cycle), showed no change in the moderately resistant line; however, it showed an increase in the susceptible line. The increase in fumarase gene expression in moderately resistant line was higher than the susceptible line. The result showed the activation of glyoxylate cycle and destruction of fatty acids in the susceptible line. CONCLUSIONS: Activation of glyoxylate cycle indicated induction of senescent symptoms by OA in susceptible line. Increasing in H2O2 leads to oxidative burst and cell death. Cell death has an apparent benefit for development and growth of necrotrophic pathogens in the plant cells. The study of resistance mechanisms in response to the pathogen is useful for breeding programs to provide lines with higher resistance to this pathogen.

Peganum harmala L.'s anti-growth effect on a breast cancer cell line.

This research was done to evaluate the induction of apoptosis in MDA-MB-231 breast cancer cell line by Peganum harmala's extract, in which a significant amount of ß-carbolines is included. The apoptosis incidence was assessed through Annexin-V-Flous kit. The expressions of genes through which intrinsic apoptosis pathway are involved, Bax, Bcl-2, Bid, and Puma, over the genes the expressions of which are linked to extrinsic apoptosis pathway, TRAIL, Caspase8, p21, and p53, were examined by RT-PCR and Real-time PCR. The results demonstrate that the extract decreases the growth rate of the cancer cell line through inducing apoptosis mechanism. As long as the expression of anti-apoptosis Bcl-2 gen reduced dramatically, an over-expression in Bax and Puma genes was monitored indicating activation of intrinsic apoptosis pathway. A notable over-expression observed with TRAIL and Caspase8 genes as well as Bid gene. The latter is an intermediate for both intrinsic and extrinsic pathways of apoptosis.