Anthony F. van Herwaarden

Genotypic variation in water-soluble carbohydrate accumulation in wheat

The water-soluble carbohydrate (WSC) that accumulates in the stems of wheat during growth can be an important contributor to grain filling, particularly under conditions when assimilation is limited, such as during end-of-season drought. WSC concentration was measured at anthesis across a diverse set of wheat genotypes over multiple environments. Environmental differences in WSC concentration were large (means for the set ranging between 108 and 203 mg g-1 dry weight), and there were significant and repeatable differences in WSC accumulation among genotypes (means ranging from 112 to 213 mg g-1 dry weight averaged across environments), associated with large broad-sense heritability (H = 0.90 ± 0.12). These results suggest that breeding for high WSC should be possible in wheat. The composition of the WSC, examined in selected genotypes, indicated that the variation in total WSC was attributed mainly to variation in the fructan component, with the other major soluble carbohydrates, sucrose and hexose, varying less. The degree of polymerisation (DP) of fructo-oligosaccharides was up to ~13 in samples where higher levels of WSC were accumulated, owing either to genotype or environment, but the higher DP components (DP > 6) were decreased in samples of lower total WSC. The results are consistent with fructan biosynthesis occurring via a sequential mechanism that is dependent on the availability of sucrose, and differences in WSC contents of genotypes are unlikely to be due to major mechanistic differences.

Molecular Dissection of Variation in Carbohydrate Metabolism Related to Water-Soluble Carbohydrate Accumulation in Stems of Wheat

Water-soluble carbohydrates (WSCs; composed of mainly fructans, sucrose [Suc], glucose [Glc], and fructose) deposited in wheat (Triticum aestivum) stems are important carbon sources for grain filling. Variation in stem WSC concentrations among wheat genotypes is one of the genetic factors influencing grain weight and yield under water-limited environments. Here, we describe the molecular dissection of carbohydrate metabolism in stems, at the WSC accumulation phase, of recombinant inbred Seri/Babax lines of wheat differing in stem WSC concentrations. Affymetrix GeneChip analysis of carbohydrate metabolic enzymes revealed that the mRNA levels of two fructan synthetic enzyme families (Suc:Suc 1-fructosyltransferase and Suc:fructan 6-fructosyltransferase) in the stem were positively correlated with stem WSC and fructan concentrations, whereas the mRNA levels of enzyme families involved in Suc hydrolysis (Suc synthase and soluble acid invertase) were inversely correlated with WSC concentrations. Differential regulation of the mRNA levels of these Suc hydrolytic enzymes in Seri/Babax lines resulted in genotypic differences in these enzyme activities. Down-regulation of Suc synthase and soluble acid invertase in high WSC lines was accompanied by significant decreases in the mRNA levels of enzyme families related to sugar catabolic pathways (fructokinase and mitochondrion pyruvate dehydrogenase complex) and enzyme families involved in diverting UDP-Glc to cell wall synthesis (UDP-Glc 6-dehydrogenase, UDP-glucuronate decarboxylase, and cellulose synthase), resulting in a reduction in cell wall polysaccharide contents (mainly hemicellulose) in the stem of high WSC lines. These data suggest that differential carbon partitioning in the wheat stem is one mechanism that contributes to genotypic variation in WSC accumulation.

Reduced growth of autumn-sown wheat in a low-P soil is associated with high colonisation by arbuscular mycorrhizal fungi

Autumn-sown wheat (Triticum aestivum) was studied over two seasons in south-eastern Australia, on a low-P soil where indigenous arbuscular mycorrhizal fungi (AMF) were known to provide little nutritional benefit to crops. It was hypothesised that AMF would be parasitic under these circumstances. Shoot dry mass and water soluble carbohydrate (WSC) reserves in roots and shoots were measured for wheat grown with or without P-fertiliser, in plots where crop sequences had produced either high or low colonisation by AMF. Application of P-fertiliser greatly increased crop growth and decreased colonisation by AMF. At tillering, colonisation by AMF ranged from 24 to 66% of root length when no P was applied and from 11 to 32% when P was applied. At each P-level, high colonisation correlated with reductions of around 20% in stem and root WSC concentrations (first season) or shoot WSC content and shoot dry mass (much drier second season). Impacts on yield were not significant (first season) or largely masked by water-stress and frost (second season). While the major fungal root diseases of the region were absent, interactions between crop sequence and other unknown biotic constraints could not be discounted. The results are consistent with the parasitic impacts of colonisation by AMF being induced primarily through the winter conditions experienced by the crops until anthesis. It is concluded that wheat in south-eastern Australia may benefit from reduced colonisation by AMF, which could achieved through selected crop sequences or, perhaps, targeted wheat breeding programs.

Plot size matters: interference from intergenotypic competition in plant phenotyping studies

Genetic and physiological studies often comprise genotypes diverse in vigour, size and flowering time. This can make the phenotyping of complex traits challenging, particularly those associated with canopy development, biomass and yield, as the environment of one genotype can be influenced by a neighbouring genotype. Limited seed and space may encourage field assessment in single, spaced rows or in small, unbordered plots, whereas the convenience of a controlled environment or greenhouse makes pot studies tempting. However, the relevance of such growing conditions to commercial field-grown crops is unclear and often doubtful. Competition for water, light and nutrients necessary for canopy growth will be variable where immediate neighbours are genetically different, particularly under stress conditions, where competition for resources and influence on productivity is greatest. Small hills and rod-rows maximise the potential for intergenotypic competition that is not relevant to a crops performance in monocultures. Response to resource availability will typically vary among diverse genotypes to alter genotype ranking and reduce heritability for all growth-related traits, with the possible exception of harvest index. Validation of pot experiments to performance in canopies in the field is essential, whereas the planting of multirow plots and the simple exclusion of plot borders at harvest will increase experimental precision and confidence in genotype performance in target environments.

Use of dry matter content as a rapid and low-cost estimate for ranking genotypic differences in water-soluble carbohydrate concentrations in the stem and leaf sheath of Triticum aestivum

Stem water-soluble carbohydrates (WSC) are an important source of temporary carbohydrate reserve in cool-season cereals. Genotypic variation in stem WSC concentration in wheat at anthesis is often positively associated with grain weight and yield in water-limited environments. In this study we have examined the relationship between dry matter content (DMC, dry weight per unit of fresh weight) and WSC concentration in field-grown bread wheat. Strong correlations (r = 0.92–0.95) were observed between DMC and WSC concentration in the stem and leaf sheath from the top two or three internodes of recombinant inbred lines from a cross between Seri M82 and Babax, at anthesis or 1 week after anthesis, in several field experiments. This strong correlation was also observed in diverse genotypes grown under rainfed or irrigated conditions. DMC and WSC concentration were also positively correlated in the whole above-ground biomass of wheat at anthesis (r = 0.74–0.91). Measurement of stem and leaf sheath DMC and WSC concentration in a small number of samples would allow the rapid prediction of WSC concentrations in a large number of field samples with reasonable accuracy, as demonstrated in a small dataset in this study. These data indicate that DMC can serve cereal breeding as a rapid and low-cost selection tool for genotypic ranking of WSC concentrations in breeding populations.

Linked gene networks involved in nitrogen and carbon metabolism and levels of water-soluble carbohydrate accumulation in wheat stems

High levels of water-soluble carbohydrates (WSC) provide an important source of stored assimilate for grain filling in wheat. To better understand the interaction between carbohydrate metabolism and other metabolic processes associated with the WSC trait, a genome-wide expression analysis was performed using eight field-grown lines from the high and low phenotypic tails of a wheat population segregating for WSC and the Affymetrix wheat genome array. The 259 differentially expressed probe sets could be assigned to 26 functional category bins, as defined using MapMan software. There were major differences in the categories to which the differentially expressed probe sets were assigned; for example, probe sets upregulated in high relative to low WSC lines were assigned to category bins such as amino acid metabolism, protein degradation and transport and to be involved in starch synthesis-related processes (carbohydrate metabolism bin), whereas downregulated probe sets were assigned to cell wall-related bins, amino acid synthesis and stress and were involved in sucrose breakdown. Using the set of differentially expressed genes as input, chemical–protein network analyses demonstrated a linkage between starch and N metabolism via pyridoxal phosphate. Twelve C and N metabolism-related genes were selected for analysis of their expression response to varying N and water treatments in the field in the four high and four low WSC progeny lines; the two nitrogen/amino acid metabolism genes demonstrated a consistent negative association between their level of expression and level of WSC. Our results suggest that the assimilation of nitrogen into amino acids is an important factor that influences the levels of WSC in the stems of field-grown wheat.