ABSTRACT
Nitrogen is one of the most important macronutrients for crop growth and metabolism. To identify marker-trait associations for complex nitrogen use efficiency (NUE)-related agronomic traits, field experiments were conducted on nested synthetic wheat introgression libraries at three nitrogen input levels across two seasons. The introgression libraries were genotyped using the 35K Axiom® Wheat Breeder's Array and genetic diversity and population structure were examined. Significant phenotypic variation was observed across genotypes, treatments, and their interactions across seasons for all the 22 traits measured. Significant positive correlations were observed among grain yield and yield-attributing traits and root traits. Across seasons, a total of 233 marker-trait associations (MTAs) associated with fifteen traits of interest at different levels of nitrogen (N0, N60, and N120) were detected using 9,474 genome-wide single nucleotide polymorphism (SNP) markers. Of these, 45 MTAs for 10 traits in the N0 treatment, 100 MTAs for 11 traits in the N60 treatment, and 88 MTAs for 11 traits in the N120 treatment were detected. We identified putative candidate genes underlying the significant MTAs which were associated directly or indirectly with various biological processes, cellular component organization, and molecular functions involving improved plant growth and grain yield. In addition, the top 10 lines based on N response and grain yield across seasons and treatments were identified. The identification and introgression of superior alleles/donors improving the NUE while maintaining grain yield may open new avenues in designing next generation nitrogen-efficient high-yielding wheat varieties.
ABSTRACT
This chapter details a reproducible method for the transformation of spring wheat using Agrobacterium tumefaciens via the direct inoculation of bacteria into immature seeds in planta as described in patent WO 00/63398 (1). Transformation efficiencies from 1 to 30% have been obtained and average efficiencies of at least 5% are routinely achieved. Regenerated plants are phenotypically normal with 30-50% of transformation events carrying introduced genes at single insertion sites, a higher rate than is typically reported for transgenic plants produced using biolistic transformation methods.
Subject(s)
Agrobacterium tumefaciens/metabolism , Gene Transfer Techniques , Transformation, Genetic , Triticum/genetics , Triticum/microbiology , Acclimatization , Plants, Genetically Modified , Seeds/genetics , Seeds/microbiology , Soil , Suspensions , Tissue Culture Techniques , Triticum/cytology , Triticum/growth & developmentABSTRACT
Pantothenate (vitamin B(5)) is the precursor of the 4'-phosphopantetheine moiety of coenzyme A and acyl-carrier protein. It is made by plants and microorganisms de novo, but is a dietary requirement for animals. The pantothenate biosynthetic pathway is well-established in bacteria, comprising four enzymic reactions catalysed by ketopantoate hydroxymethyltransferase (KPHMT), L: -aspartate-alpha-decarboxylase (ADC), pantothenate synthetase (PS) and ketopantoate reductase (KPR) encoded by panB, panD, panC and panE genes, respectively. In higher plants, the genes encoding the first (KPHMT) and last (PS) enzymes have been identified and characterised in several plant species. Commercially, pantothenate is chemically synthesised and used in vitamin supplements, feed additives and cosmetics. Biotransformation is an attractive alternative production system that would circumvent the expensive procedures of separating racemic intermediates. We explored the possibility of manipulating pantothenate biosynthesis in plants. Transgenic oilseed rape (Brassica napus) lines were generated in which the E. coli KPHMT and PS genes were expressed under a strong constitutive CaMV35SS promoter. No significant change of pantothenate levels in PS transgenic lines was observed. In contrast plants expressing KPHMT had elevated pantothenate levels in leaves, flowers siliques and seed in the range of 1.5-2.5 fold increase compared to the wild type plant. Seeds contained the highest vitamin content, indicating that they might be the ideal target for production purposes.
Subject(s)
Brassica rapa/metabolism , Genetic Engineering , Pantothenic Acid/metabolism , Brassica rapa/genetics , Escherichia coli/enzymology , Escherichia coli/genetics , Fatty Acids/analysis , Flowers/metabolism , Gene Expression Regulation, Plant , Genes, Bacterial , Glucuronidase/metabolism , Hydroxymethyl and Formyl Transferases/genetics , Hydroxymethyl and Formyl Transferases/metabolism , Pantothenic Acid/biosynthesis , Pantothenic Acid/isolation & purification , Phenotype , Plant Leaves/metabolism , Plants, Genetically Modified , Plasmids/genetics , Seedlings/metabolism , Seeds/metabolismABSTRACT
Foods high in resistant starch have the potential to improve human health and lower the risk of serious noninfectious diseases. RNA interference was used to down-regulate the two different isoforms of starch-branching enzyme (SBE) II (SBEIIa and SBEIIb) in wheat endosperm to raise its amylose content. Suppression of SBEIIb expression alone had no effect on amylose content; however, suppression of both SBEIIa and SBEIIb expression resulted in starch containing >70% amylose. When the >70% amylose wheat grain was fed to rats in a diet as a wholemeal, several indices of large-bowel function, including short-chain fatty acids, were improved relative to standard wholemeal wheat. These results indicate that this high-amylose wheat has a significant potential to improve human health through its resistant starch content.
