Mineral deficiency limits crop production in most soils and in Asia alone, about 50% of rice lands are phosphorous deficient. In an attempt to determine the mechanism of rice adaptation to phosphorous deficiency, changes in proteome patterns associated with phosphorous deficiency have been investigated. We analyzed the parental line Nipponbare in comparison to its near isogenic line (NIL6-4) carrying a major phosphorous uptake QTL (Pup1) on chromosome 12. Using 2-DE, the proteome pattern of roots grown under 1 and 100 microM phosphorous were compared. Out of 669 proteins reproducibly detected on root 2-DE gels, 32 proteins showed significant changes in the two genotypes. Of them, 17 proteins showed different responses in two genotypes under stress condition. MS resulted in identification of 26 proteins involved in major phosphorous deficiency adaptation pathways including reactive oxygen scavenging, citric acid cycle, signal transduction, and plant defense responses as well as proteins with unknown function. Our results highlighted a coordinated response in NIL in response to phosphorous deficiency which may confer higher adaptation to nutrient deficiency.
Eugenol is an aromatic component of clove oil that has therapeutic potential as an antifungal drug, although its mode of action and precise cellular target(s) remain ambiguous. To address this knowledge gap, a chemical-genetic profile analysis of eugenol was done using ∼4700 haploid Saccharomyces cerevisiae gene deletion mutants to reveal 21 deletion mutants with the greatest degree of susceptibility. Cellular roles of deleted genes in the most susceptible mutants indicate that the main targets for eugenol include pathways involved in biosynthesis and transport of aromatic and branched-chain amino acids. Follow-up analyses showed inhibitory effects of eugenol on amino acid permeases in the yeast cytoplasmic membrane. Furthermore, phenotypic suppression analysis revealed that eugenol interferes with two permeases, Tat1p and Gap1p, which are both involved in dual transport of aromatic and branched-chain amino acids through the yeast cytoplasmic membrane. Perturbation of cytoplasmic permeases represents a novel antifungal target and may explain previous observations that exposure to eugenol results in leakage of cell contents. Eugenol exposure may also contribute to amino acid starvation and thus holds promise as an anticancer therapeutic drug. Finally, this study provides further evidence of the usefulness of the yeast Gene Deletion Array approach in uncovering the mode of action of natural health products.
Wild relatives of wheat serve as an extraordinary source of variability for breeding programs due to their capabilities to respond to various environmental stresses. Here, we investigated some species possessing a D genome (T. aestivum, Ae. tauschii, Ae. crassa and Ae. cylindrica) in terms of relative water content (RWC), stomatal conductance (Gs), relative chlorophyll content, initial fluorescence (Fo), maximum quantum yield of PSII (Fv/Fm), maximum primary yield of PSII photochemistry (Fv/Fo), as well as shoot fresh and dry biomasses under control and water deficit conditions. Our results revealed that water deficit negatively affected all traits; shoot fresh weight, Gs and RWC showed the highest reduction compared to the control condition. Principal component analysis (PCA) identified two PCs that accounted for 53.36% of the total variation in the water deficit conditions. Correlation analysis and PCA-based biplots showed that stress tolerance index (STI) is significantly associated with Fv/Fm and Fv/Fo under water stress conditions, suggesting that these are the best parameters to evaluate when screening for tolerant samples at the seedling stage. We identified 19 accessions from Ae. crassa and one from Ae. tauschii as the most tolerant samples. In conclusion, Ae. crassa might provide an ideal genetic resource for drought-tolerant wheat breeds.
Dorema ammoniacum D. Don. (Apiaceae), a native medicinal plant in Iran, is classified as a vulnerable species. Root, hypocotyl, and cotyledon segments were cultured on Murashige and Skoog (MS) (1962) medium supplemented with either 2,4-dichlorophenyoxyacetic acid (2,4-D) or naphathalene acetic acid (NAA), at 0-2 mg l -1 , alone or in combination with either benzyladenine (BA) or kinetin (KN), at 0-2 mg l -1 for callus induction. The best response (100%) was observed from root segments on MS medium containing 1 mg l -1 NAA and 2 mg l -1 BA. The calli derived from various explants were subcultured on MS medium supplemented with BA (1-4 mg l -1 ) alone or in combination with NAA or indole-3-butyric acid (IBA), at 0.2 or 0.5 mg l -1 for shoot induction. Calli derived from hypocotyl segments showed significantly higher frequency of plantlet regeneration and number of plantlets than the calli derived from root and cotyledon segments. Therefore, MS medium supplemented with 2 mg l -1 BA and 0.2 mg l -1 IBA produced the highest frequency of shoot regeneration (87.3%) in hypocotyl-derived callus. The optimal medium for rooting contained 2.5 mg l -1 IBA on which 87.03% of the regenerated shoots developed roots with an average number of 5.2 roots per shoots within 30 days. These plantlets were hardened and transferred to the soil. The described method can be successfully employed for the large-scale multiplication and conservation of germplasm this plant.
In the last decades, extensive research on the effects of nano-TiO₂ on plant systems and different microorganisms has confirmed its photocatalytic and antimicrobial activity. However, there is no report on its application in plant cell and tissue culture as well as its role in eliminating contaminating microorganisms in tissue culture. In this work, barley mature embryos were cultured in Murashige and Skoog medium with four concentrations (0, 10, 30, 60 μg/ml) of TiO₂ suspension in four repetitions. Quantitative and qualitative characteristics of calli were analyzed after each subculture. Data analysis for calli number in the first culture and callus size in all three cultures showed that the effect of treatment was significant at p > 0.95. As a result, quantitative features such as callus color, shape, embryogenesis, etc. were completely similar in both control and TiO₂ nanoparticle treatments; there is no doubt that TiO₂ nanoparticles could dramatically increase callugenesis and the size of calli. As well, TiO₂ nanoparticles are effective bactericides with an aseptic effect, causing no negative change in the quality of the callus. It is necessary to do more complementary works to identify mechanisms involved for the increased calli size and embryogenesis of explants in darkness.
The aim of the present study was to investigate the relationship between the presence of the meiotic spindle and zona pellucida (ZP) birefringence with morphology of in vivo- and in vitro-matured human oocytes. Germinal vesicles (n=47) and MI (n=38) oocytes obtained from stimulated ovaries of patients undergoing intracytoplasmic sperm injection (ICSI) underwent IVM. Using a PolScope (OCTAX PolarAID; Octax, Herbon, Germany), the presence of spindles and ZP birefringence was assessed in both in vivo-matured (n=56) and IVM (n=56) oocytes. In addition, the morphology of each matured oocyte was evaluated microscopically. There were insignificant differences for ZP birefringence and meiotic spindle between the in vivo-matured and IVM MII oocytes. Subanalysis revealed that the rates of morphologically abnormal oocytes did not differ significantly between the two groups, except in the case of irregular shape (P=0.001), refractile body (P=0.001) and fragmented polar body (P=0.03), which were higher in IVM oocytes. In the case of in vivo-matured oocytes, a significantly higher percentage of oocytes with intracytoplasmic and both intra- and extracytoplasmic abnormalities have a low birefringent ZP (P=0.007 and P=0.02, respectively). There was no relationship between morphological abnormalities and spindle detection. The findings suggest that clinical IVM is a safe technology that maintains the high maturation rate and integrity of oocytes. In addition, the use of the non-invasive PolScope is recommended for the detection of oocytes most suitable for ICSI.
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