Applied biotechnology to improve Mediterranean rice varieties = Biotecnologia aplicada a la millora de varietats d’arròs mediterrànies

Abstract

[eng] The current world population is over 7.4 billion and expected to exceed 9 billion in 2040, causing a 70% increase in food demand. Global environmental degradation, in the form of salinization, pollution and global warming, has also reduced the availability of suitable arable land and water sources, contributing to promote crop improvement in order to increase the potential yields. Rice (Oryza sativa) is the most widely consumed staple food for a large proportion of the world population. Classical rice breeding programs use its natural variability to create new allelic combinations which are screened for selecting those presenting superior agronomic traits such as improved yield. Those improved lines are stabilized through inbreeding to maintain the phenotype in their progeny. Certified seed producers systematically select and propagate registered varieties year by year in order to maintain their uniformity and the original registered cultivar traits, since natural mutations, spontaneous breeding between varieties and alien grain contamination can introduce undesirable variability at this stage. Nowadays biotechnology is used to drive the improvement of rice traits such as increased yield and grain quality. Moreover it helps to rapidly bestow tolerance to biotic (diseases and insects) and abiotic (drought, salinity, cold temperatures, nutrients deficiency) factors. Some of the available biotechnological techniques applied for crop improvement are i) the genetic engineering, which allows the addition of foreign genes in the rice genome although being controversial due to the social and environmental concerns, ii) the anther culture, which fasten and improves the selection of new breeding lines, and iii) the Targeting Induced Local Lesions IN Genomes (TILLING) which combines the production of large mutant populations with the detection of mutants in genes of interest through molecular screening. The main aim of the thesis is to study different biotechnological tools and their applications to the improvement of Mediterranean rice varieties. To achieve this biotechnology is used to study the pollen dispersion of a genetically engineered rice line, to accelerate the stabilization process through anther culture technique and to introduce new variability using a mutagenesis protocol followed by molecular detection of mutants. In this thesis we first studied the pollen-mediated gene flow between wild rice, conventional rice and an herbicide resistant transgenic rice line in order to determine gene flow rates in relation to the distance and the prevailing wind speed and direction. Results showed that pollen dispersal is dramatically effected by the distances between rice plants and the speed and direction of the prevailing wind. Furthermore, the enhanced pollen dispersal capability of weedy rice can also play an important role in transgenic pollen dispersal, which unfortunately had been underestimated. Then, we adapted an anther culture protocol in order to efficiently obtain commercial dihaploid lines from a Mediterranean japonica variety. Furthermore, we described the greenhouse and field trials used to select the best lines for registration which are now being successfully commercialized. Finally, we developed a fast protocol for obtaining mutants with agronomic interest. This protocol is based on ethyl methanesulfonate mutagenesis of seed-derived calli. The in vitro regenerated mutant population plants were directly screened for senescence-related genes, allowing to shorten in more than eight months the common seed mutagenesis protocol. The molecular screening protocol was also optimized and several potential delayed senescence mutants were identified and tested