RESUMEN
Although the SnRK2 class of Ser/Thr protein kinases is critical for signal transduction and abiotic stress resistance in plants, there have been no studies to examine SnRK2 in Jatropha curcas L. In the present study, JcSnRK2 was cloned from J. curcas using the rapid amplification of cDNA end technique and characterized. The JcSnRK2 genomic sequence is 2578 base pairs (bp), includes 10 exons and 9 introns, and the 1017-bp open reading frame encodes 338 amino acids. JcSnRK2 was transcribed in all examined tissues, with the highest transcription rate observed in the roots, followed by the leaves and stalks; the lowest rate was observed in flowers and seeds. JcSnRK2 expression increased following abscisic acid treatment, salinity, and drought stress. During a 48-h stress period, the expression of JcSnRK2 showed 2 peaks and periodic up- and downregulation. JcSnRK2 was rapidly activated within 1 h under salt and drought stress, but not under cold stress. Because of the gene sequence and expression similarity of JcSnRK2 to AtSnRK2.8, primarily in the roots, an eukaryotic expression vector containing the JcSnRK2 gene (pBI121-JcSnRK2) was constructed and introduced to the Arabidopsis AtSnRK2.8 mutant snf2.8. JcSnRK2-overexpressing plants exhibited higher salt and drought tolerance, further demonstrating the function of JcSnRK2 in the osmotic stress response. J. curcas is highly resistant to extreme salt and drought conditions and JcSnRK2 was found to be activated under these conditions. Thus, JcSnRK2 is potential candidate for improving crop tolerance to salt and drought stress.
Asunto(s)
Clonación Molecular , Jatropha/genética , Proteínas de Plantas/genética , Proteínas Serina-Treonina Quinasas/genética , Arabidopsis/genética , Arabidopsis/crecimiento & desarrollo , Evolución Molecular , Flores/genética , Regulación de la Expresión Génica de las Plantas , Mutación , Filogenia , Hojas de la Planta/genética , Raíces de Plantas/genética , Plantas Modificadas Genéticamente/genética , Plantas Modificadas Genéticamente/crecimiento & desarrollo , Tolerancia a la Sal , Semillas/genética , Estrés FisiológicoRESUMEN
The expression of the inducible nitric oxide synthase enzyme (iNOS) is a response to an inflammatory stimulus and produces a large amount of nitric oxide (NO), which may act as a cytotoxic molecule against the invading microorganism and may be related to both harmful and beneficial effects to tissues. OBJECTIVE AND MATERIAL AND METHODS: In order to further characterize the presence of NO in human periodontal disease, we undertook a quantitative study of iNOS positive cells in samples of clinically healthy gingival tissues, plaque-induced gingivitis and localized chronic periodontitis using immunohistochemistry. RESULTS: A significant increase in the number of iNOS+ cells mm-2 was found in the samples of the gingivitis and periodontitis compared with those of the control. In all groups most of the polymorphonuclear cells showed intense immunoreactivity for iNOS independent of the disease stage, and the percentage of iNOS+ polymorphonuclear cells increased significantly in periodontal disease when compared with the control. CONCLUSION: Our results indicate that iNOS increases in the presence of periodontal disease. In addition, our findings suggest that polymorphonuclear cells present an additional activation pathway in periodontal disease, expressing significant iNOS and probably representing an important source of NO in human periodontal disease that has not been previously reported.