Physiological characterization and gene expression profiling of sugarcane (Saccharum spp) genotypes contrasting for drought tolerance

Author(s)

João Felipe Nebó Carlos de Oliveira

• Document type: Doctoral Thesis
• Press: Piracicaba.
• Institution: Universidade de São Paulo (USP). Escola Superior de Agricultura Luiz de Queiroz (ESALA/BC)
• Defense date: 2013-02-01

Examining board members

Antonio Vargas de Oliveira Figueira; Marcos Silveira Buckeridge; Denis Ubeda de Lima; Renato Vicentini dos Santos; Silvana Aparecida Creste Dias de Souza

• Advisor: Antonio Vargas de Oliveira Figueira

Abstract

Sugarcane (Saccharum spp.) is a major crop in Brazil as feedstock for the sugar and ethanol industries. The Brazilian sugarcane industry has been expanding rapidly due to increased demand for biofuels. To attend the increasing ethanol demand and secure food production, sugarcane production must expand the cultivated area, incorporating land from \'cerrado\' and degraded pastures in Southeast and Western Brazil, areas characterized by a dry winter with a prolonged water deficit period. Drought-tolerant cultivars with higher yield potentials in such regions are desired for this expansion, and the elucidation of the molecular and physiological basis of drought tolerance in sugarcane is valuable for developing such cultivars. To investigate physiological and molecular mechanisms involved in tolerance to water deficit, different approaches based on field and greenhouse trials were evaluated, mainly focusing on phenotyping and gene expression profiling. The addition of polyethylene glycol (PEG) as a stress inducer in nutrient solution was also evaluated to simulate droughtresponse in hydroponically-grown sugarcane, as well as the solution\'s electrolyte leakage after exposure of leaf segments to methyl-viologen (Paraquat). Two genotypes previously selected were used to test the hypothesis of the mechanisms underlying on the differential response to drought, due to their behavior as contrasting genotypes, where \'IACPSP 94-2094\' was tested as drought-tolerant and \'IACSP 97-7065\' as drought-sensitive. When plants were hydroponically grown on solution containing 15% PEG, genotypic differences for leaf water potential and gas exchange were apparent after 72 h, whereas the improved oxidative response of the tolerant genotype after leaf-disk treatment with methyl-viologen suggested the potential of this approach to be used for early drought-tolerance screening. Ten genotypes including the two previously selected were evaluated for drought tolerance on field conditions at \'Jalles Machado\' sugarmill (Goianésia, GO), a drought-prone location. \'IACSP 94-2094\' showed enhanced features for drought tolerance (early stomatal closure; maintenance of leaf water potential and photochemical adjustment), absent in \'IACSP 97-7065\'. Molecular studies (microarrays, RNAseq and RT-qPCR) were also conducted using these genotypes, aiming to identify drought-resposive genes and confirm their roles on tolerance. From a set of 14,522 genes in a microarray chip, 91 (~0.63%) were differentially expressed between irrigated and non-irrigated treatment during early drought, whereas 576 (~4%) showed differential expression during severe drought between water treatments, including many genes traditionally associated with drought response. Samples from these same genotypes grown in greenhouse conditions under irrigation and water deprivation were used for gene expression profiling by RNAseq. From a sugarcane reference dataset with 43,141 genes, 2,300 (5.3%) genes were identified as differentially expressed, including novel and undescribed genes that might play a role in drought tolerance. Most genes identified as differentially expressed on microarrays were also observed for RNAseq, with high correlation between both approaches (0.78 and 0.84 for tolerant and sensitive genotypes, respectively). After validation, genes related to drought response have been used for functional genomics studies aiming the development of new cultivars tolerant to drought, either through conventional or molecular breeding. (AU)

• FAPESP's process: 08/06446-2 - Characterization of gene expression profile of sugarcane-contrasting cultivars for drought stress
• Grantee: João Felipe Nebó Carlos de Oliveira
• Support Opportunities: Scholarships in Brazil - Doctorate (Direct)