ABSTRACT. Gene expression related to drought response in the leaf tissues of two Brazilian upland cultivars, the drought-tolerant Douradão and the drought-sensitive Primavera, was analyzed. RNAseq identified 27,618 transcripts in the Douradão cultivar, with 24,090 (87.2%) homologous to the rice database, and 27,221 transcripts in the Primavera cultivar, with 23,663 (86.9%) homologous to the rice database. Gene-expression analysis between control and water-deficient treatments revealed 493 and 1154 differentially expressed genes in Douradão and Primavera cultivars, respectively. Genes exclusively expressed under drought were identified for Douradão, including two genes of particular interest coding for the protein peroxidase precursor, which is involved in three distinct metabolic pathways. Comparisons between the two drought-exposed cultivars revealed 2314 genes were R.D.D. Silveira et al. 8182©FUNPEC-RP www.funpecrp.com.br Genetics and Molecular Research 14 (3): 8181-8200 (2015) differentially expressed (978 upregulated, 1336 downregulated in Douradão). Six genes distributed across 4 different transcription factor families (bHLH, MYB, NAC, and WRKY) were identified, all of which were upregulated in Douradão compared to Primavera during drought. Most of the genes identified in Douradão activate metabolic pathways responsible for production of secondary metabolites and genes coding for enzymatically active signaling receptors. Quantitative PCR validation showed that most gene expression was in agreement with computational prediction of these transcripts. The transcripts identified here will define molecular markers for identification of Cisacting elements to search for allelic variants of these genes through analysis of polymorphic SNPs in GenBank accessions of upland rice, aiming to develop cultivars with the best combination of these alleles, resulting in materials with high yield potential in the event of drought during the reproductive phase.

This work aimed to evaluate the drought tolerance of transformed plants of the cultivar BRSMG Curinga that overexpress the rice phospholipase D α1 (OsPLDα1) gene. The productivity of independent transformation event plants of the OsPLDα1 gene was evaluated in an experiment where 19 days of water deficit were applied at the reproductive stage, a very strict growing condition for upland rice. The non-genetically modified cultivar (NGM) under drought treatment reduced productivity by 89% compared with that under irrigated treatment, whereas transformed plants (PLDα1_E2) reduced productivity by only 41%. After the drought treatment, the PLDα1_E2 plants productivity was five times greater than that of the NGM plant. Moreover, no adverse effects on growth and development of the transgenic plants were observed. Seven days after the resumption of irrigation, PLDα1_E2 plants had higher stomatal conductance, greater photosynthetic rate, and transpiration rate than did NGM plants, as well as a higher expression level of the OsPLDα1 gene. A delay in the senescence process was observed in these PLDα1_E2 plants, and this was determined for the recovery of photosynthesis, with greater expression of the Rubisco and lower expression of the SOD. This finding was suggestive of decreased oxidative stress, probably due to gas exchange by the partial closure of the stomata of these transformed plants, which prevented the formation of reactive oxygen species. OsPLDα1 gene overexpression resulted in a reduction in production loss under severe water deficit and revealed a possibility for the development of upland rice cultivars that are more tolerant to extreme drought conditions.

The identification and validation of candidate genes related to traits of interest is a time consuming and expensive process and the homology among genes from different species can facilitate the identification of genes of the target species from the genomic information of a model species. This study aimed to quantify the expression of homologous rice genes previously related to drought tolerance in Arabidopsis. Five genes (CPK6, PLDα, GluR2, CesA8, and EIN2) were identified in rice by the homology of the amino acid sequence between rice and Arabidopsis. The genotypes Douradão (drought tolerant) and Primavera (drought susceptible) were subjected to a water deficit experiment, and subsequently evaluated for gene expression by qPCR for the five homologous and Lsi1 genes. The qPCR analysis clearly showed that the five homologous genes were expressed in rice, which is an indication that these genes could preserve their function in rice as a response to drought. In Douradão, of the five homologous genes, all but OsGluR2 displayed an increase in the average expression in drought treatment when compared to the control, while in Primavera, the average expression of the five genes did not differ between the control and drought treatment. In Douradão, the OsPLDα1, which showed the higher expression level in drought in relation to the control (10.82 times), significantly increased the gene expression in the leaf and root tissues as a response to drought, in both vegetative and reproductive stages, whereas in Primavera, this gene was suppressed in both tissues and stages under drought. Therefore, the OsPLDα1 gene was the most important in relation to drought response and is an interesting candidate for further studies in developing rice cultivars that are more tolerant to this stress.

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