Induction of the expression of defence genes in Carica papaya fruit by methyl jasmonate and low temperature treatments

Abstract The defence mechanisms that are activated by methyl jasmonate (MJ) in fruits are not well understood. In this work, we studied the expression of defence genes in papaya fruit that are induced by the exposure to MJ and/or low temperatures. The papaya fruits \u2018Maradol\u2019 were randomly divided into two groups: one group was the untreated control and the other was treated with 10-4 M of MJ. Half of the fruits from each of the two groups were stored after treatment for 5 days at 5\ubaC and 2 days at 20\ubaC. We studied the expression levels of the pdf1.1 and pdf1.2 genes by amplification from expression libraries created from the pulp and skin tissues of the papaya fruit. As a reference, the mRNA level of the 18s ribosomal gene was used. In the skin tissue, the expression levels of the pdf1.1 and pdf1.2 genes were higher immediately after MJ treatment compared to the control. Furthermore, the expression of pdf1.2 remained high after MJ treatment and subsequent storage compared to the control. It was therefore concluded that the activation of the pdf1.1 and pdf1.2 genes forms part of the molecular defence mechanism in fruits that is activated by exposure to MJ. To our knowledge, this is the first study that analyzes the gene expression in papaya fruit that is induced by the exogenous application of methyl jasmonate and cold treatment

Total RNA quality of lyophilized and cryopreserved dormant grapevine buds

Background: Plant tissues must be preserved in their collection state, especially for genome-wide expression profile studies. Lyophilization is a feasible, affordable tool when fresh tissues cannot be shipped at ultralow temperatures from their origin to the place of analysis. In this study, the total RNA quality of dormant grapevine buds ( Vitis vinifera L. cv. \u2018Flame Seedless\u2019) of freeze-dried samples stored at room temperature conditions was evaluated and compared to that of cryopreserved (-80\ub0C) grapevine buds. Results: Good yield and quality of RNA were obtained from freeze-dried dormant buds stored at room temperature for 0, 3 and 6 weeks after they were lyophilized. Further experiments confirmed that the extracted total RNA could be used for actin and \u3b2-tubulin PCR gene amplification. Conclusion: High-quality RNA that is useful for downstream applications was obtained from freeze-dried dormant grapevine bud tissue, similarly to the RNA obtained from cryopreserved dormant grapevine buds