Molecular insights into how a deficiency of amylose affects carbon allocation--carbohydrate and oil analyses and gene expression profiling in the seeds of a rice waxy mutant.

BACKGROUND: Understanding carbon partitioning in cereal seeds is of critical importance to develop cereal crops with enhanced starch yields for food security and for producing specified end-products high in amylose, ß-glucan, or fructan, such as functional foods or oils for biofuel applications. Waxy mutants of cereals have a high content of amylopectin and have been well characterized. However, the allocation of carbon to other components, such as ß-glucan and oils, and the regulation of the altered carbon distribution to amylopectin in a waxy mutant are poorly understood. In this study, we used a rice mutant, GM077, with a low content of amylose to gain molecular insight into how a deficiency of amylose affects carbon allocation to other end products and to amylopectin. We used carbohydrate analysis, subtractive cDNA libraries, and qPCR to identify candidate genes potentially responsible for the changes in carbon allocation in GM077 seeds. RESULTS: Carbohydrate analysis indicated that the content of amylose in GM077 seeds was significantly reduced, while that of amylopectin significantly rose as compared to the wild type BP034. The content of glucose, sucrose, total starch, cell-wall polysaccharides and oil were only slightly affected in the mutant as compared to the wild type. Suppression subtractive hybridization (SSH) experiments generated 116 unigenes in the mutant on the wild-type background. Among the 116 unigenes, three, AGP, ISA1 and SUSIBA2-like, were found to be directly involved in amylopectin synthesis, indicating their possible roles in redirecting carbon flux from amylose to amylopectin. A bioinformatics analysis of the putative SUSIBA2-like binding elements in the promoter regions of the upregulated genes indicated that the SUSIBA2-like transcription factor may be instrumental in promoting the carbon reallocation from amylose to amylopectin. CONCLUSION: Analyses of carbohydrate and oil fractions and gene expression profiling on a global scale in the rice waxy mutant GM077 revealed several candidate genes implicated in the carbon reallocation response to an amylose deficiency, including genes encoding AGPase and SUSIBA2-like. We believe that AGP and SUSIBA2 are two promising targets for classical breeding and/or transgenic plant improvement to control the carbon flux between starch and other components in cereal seeds.

Mapping of quantitative trait loci for fiber and lignin contents from an interspecific cross Oryza sativa×Oryza rufipogon.

Rice straw is always regarded as a by-product of rice production, but it could be a significant energy source for ruminant animals. Knowledge of the genetic variation and genetic architecture of cell wall traits will facilitate rice breeders by improving relevant traits through selective breeding and genetic engineering. The common wild rice, Oryza rufipogon Griff., which is considered to be the progenitor of Oryza sativa, has been widely utilized for the identification of genes of agronomic importance for rice genetic improvement. In the present study, the mapping of quantitative trait loci (QTLs) for acid detergent fiber (ADF), neutral detergent fiber (NDF), acid detergent lignin (ADL), and ADL/NDF ratio was carried out in two environments using a backcrossed inbred line (BIL) population derived from a cross between the recurrent parent Xieqingzao B (XB) and an accession of Dongxiang wild rice (DWR). The results indicated that all four traits tested were continuously distributed among the BILs, but many BILs showed transgressive segregation. A total of 16 QTLs were identified for the four traits, but no QTLs were in common in two environments, suggesting that environment has dramatic effects on fiber and lignin syntheses. Compared to the QTL positions for grain yield-related traits, there were no unfavorable correlations between grain yield components and cell wall traits in this population. The QTLs identified in this study are useful for the development of dual-purpose rice varieties that are high in grain yield and are also high in straw quality.

[Genetic analysis and mapping of high-tillering and dwarf mutant htd1-2 in rice].

Tillering is one of the most important agronomic traits of rice. In order to explore the molecular mechanism of rice tillering, a high-tillering dwarf 1-2 (htd1-2) mutant was isolated from the offspring of the indica rice (Oryza sativa L. ssp. indica) variety 9311 treated with 350Gy 60Co gamma-radiation. Genetic analysis showed that both high tillering and dwarf phenotypes of htd1-2 were controlled by a single recessive nuclear gene. By means of molecular marker technique, the HIGH-TILLERING DWARF1-2(HTD1-2) gene was mapped between two CAPS markers A6 and E2 on chromosome 4 within 116 kilobases. Within this region, there is a cloned gene HIGH-TILLERING DWARF1(HTD1), which controls rice tillering. By comparing sequences of HTD1 between htd1-2 and 9311, in combination with the results from dCAPS analysis, we believed that HTD1 is an orthologue of the gene HTD1-2. Because of different genetic backgrounds, htd1 and htd1-2 have different phenotypes although they are the allelic mutants. Furthermore, removal of axillary buds proved that dwarfism of htd1-2 mutant is partly attributed to its excessive tillers.

Analysis of genotype x environment interaction effects for starch pasting viscosity characteristics in Indica rice.

Milled rice (Oryza sativa L.) is composed of approximately 90% starch. The properties of starch have considerable effects on cooked rice palatability and consumer acceptability. Starch pasting viscosity parameters serve as important indices in the estimation of eating, cooking, and processing qualities of rice. In the present study, four cytoplasmic male-sterile (CMS) lines and eight restorer (R) lines have been used in an incomplete diallel cross to analyze seed effects, cytoplasmic effects, maternal gene effects, and their genotype x environment (GE) effects on the following starch pasting viscosity parameters: breakdown (BD), consistency (CS), and setback (SB). The results demonstrated that the total main genetic variances (V(G)) accounted for over 64% of the total genetic variance (V(G) +V(GE)) for the three traits, indicating that these traits were mainly controlled by the main genetic effects in addition to the GE interaction effects. The estimated total narrow-sense heritability were 67.8%, 79.5%, and 79.5% for BD, CS, and SB, respectively. The general heritability (h(2)(G)) accounted for over 75% of the total heritability (h(2)(G) +h(2)(GE)), indicating that early selection would be effective for those traits and the selection efficiencies were relatively stable in different environments.

[Recent advances in molecular mapping and cloning of useful genes from wild rice and their application in breeding].

Wild rice has adapted to weather and unfavorable environments under natural selection. It has been well recognized as a natural gene bank that conserves a lot of specific genes presently not available for extinct in the cultivated rice. There is an urgent need to explore these specific genes. The present paper summarized current researches in molecular mapping and cloning of useful genes from wild rice,and their potential application in breeding.