An autoactive NB-LRR gene causes Rht13 dwarfism in wheat.

Semidwarfing genes have greatly increased wheat yields globally, yet the widely used gibberellin (GA)-insensitive genes Rht-B1b and Rht-D1b have disadvantages for seedling emergence. Use of the GA-sensitive semidwarfing gene Rht13 avoids this pleiotropic effect. Here, we show that Rht13 encodes a nucleotide-binding site/leucine-rich repeat (NB-LRR) gene. A point mutation in the semidwarf Rht-B13b allele autoactivates the NB-LRR gene and causes a height reduction comparable with Rht-B1b and Rht-D1b in diverse genetic backgrounds. The autoactive Rht-B13b allele leads to transcriptional up-regulation of pathogenesis-related genes including class III peroxidases associated with cell wall remodeling. Rht13 represents a new class of reduced height (Rht) gene, unlike other Rht genes, which encode components of the GA signaling or metabolic pathways. This discovery opens avenues to use autoactive NB-LRR genes as semidwarfing genes in a range of crop species, and to apply Rht13 in wheat breeding programs using a perfect genetic marker.

Multiple loci with cumulative effects on late maturity α-amylase (LMA) in wheat.

MAIN CONCLUSION: The cumulative action of combinations of alleles at several loci on the wheat genome is associated with different levels of resistance to late maturity α-amylase in bread wheat. Resistance to late maturity α-amylase (LMA) in bread wheat (Triticum aestivum L.) involves a complex interaction between the genotype and the environment. Unfortunately, the incidence and severity of LMA expression is difficult to predict and once the trait has been triggered an unacceptably low falling number, high grain α-amylase may be the inevitable consequence. Wheat varieties with different levels of resistance to LMA have been identified but whilst some genetic loci have been reported, the mechanisms involved in resistance and the interaction between resistance loci requires further research. This investigation was focused on mapping resistance loci in populations derived by inter-crossing resistant wheat varieties or crossing resistant lines with a very susceptible line and then mapping quantitative trait loci. In addition to the previously reported locus on chromosome 7B for which a candidate gene has been proposed, loci were mapped on chromosomes 1B, 2A, 2B, 3A, 3B, 4A, 6A and 7D. These loci have limited effects on their own but have a cumulative effect in combination with each other. Further research will be required to determine the nature of the causal genes at these loci, to develop diagnostic markers and determine how the genes fit into the pathway that leads to the induction of α-AMY1 transcription in the aleurone of developing wheat grains. Depending on the target environmental conditions, different combinations of alleles may be required to achieve a low risk of LMA expression.

Gibberellins in developing wheat grains and their relationship to late maturity α-amylase (LMA).

MAIN CONCLUSION: α-Amylase synthesis by wheat aleurone during grain development (late maturity α-amylase) appears to be independent of gibberellin unlike α-amylase synthesis by aleurone during germination or following treatment with exogenous GA. Late-maturity α-amylase (LMA) in wheat (Triticum aestivum L.) involves the synthesis of α-amylase by the aleurone tissue during grain development. Previous research identified a putative ent-copalyl diphosphate synthase gene, coding for an enzyme that controls the first step in gibberellin biosynthesis, that underlies the major genetic locus involved in variation in LMA phenotype. The reported results for gene transcript analysis, preliminary gibberellin analysis and the effects of DELLA mutants on LMA phenotype appeared to be consistent with involvement of gibberellin but did not provide definitive proof of a causal link. Conversely, several observations do not appear to be consistent with this hypothesis. In this current study, LMA phenotype, gibberellin profiles and ABA content were recorded for experiments involving susceptible and resistant genotypes, gibberellin biosynthesis inhibitors, genetic lines containing different LMA quantitative trait loci and treatment of distal halves of developing grains with exogenous gibberellin. The results suggested that gibberellin may not be a prerequisite for LMA expression and further that the mechanism involved in triggering α-amylase synthesis did not correspond to the model proposed for germination and gibberellin challenged aleurone of ripe grain. The results provide new insight into LMA and highlight the need to investigate alternate pathways for the induction of α-amylase gene transcription, the function of novel 1-ß-OH gibberellins and other functions of DELLA proteins in developing grains.

Late-maturity α-amylase expression in wheat is influenced by genotype, temperature and stage of grain development.

MAIN CONCLUSION: Late-maturity α-amylase (LMA) expression in wheat grains can be induced by either a cool temperature shock close to physiological maturity or continuous cool maximum temperatures during grain development. Late-maturity α-amylase (LMA) is a genetic trait in wheat (Triticum aestivum L.) involving the production of α-amylase during grain development, which can result in an unacceptably low Falling Number (FN) in mature grain and consequent grain downgrading. Comparison of the FN test, an α-amylase activity assay and a high pI α-amylase-specific ELISA on the same meal samples gave equivalent results; ELISA was used for further experiments because of its isoform specificity. A cool temperature shock during the middle stages of grain development is known to induce LMA and is used for phenotypic screening. It was determined that a cool temperature treatment of seven days was required to reliably induce LMA. Glasshouse studies performed in summer and winter demonstrated that temperature affected the timing of sensitivity to cool-shock by altering the rate and duration of grain development, but that the sensitive grain developmental stage was unchanged at 35-45% moisture content. Wheat varieties with Rht-B1b or Rht-D1b dwarfing genes responded to a cool-shock only from mid grain filling until physiological maturity, whilst genotypes with Rht8c or without a dwarfing gene expressed LMA in response to a cool-shock during a wider developmental range. A continuous cool maximum temperature regimen (23 °C/15 °C day/night) during grain development also resulted in LMA expression and showed a stronger association with field expression than the cool-shock treatment. These results clarify how genotype, temperature and grain developmental stage determine LMA expression, and allow for the improvement of LMA phenotypic screening methods.

Identification of differentially senescing mutants of wheat and impacts on yield, biomass and nitrogen partitioning.

Increasing photosynthetic capacity by extending canopy longevity during grain filling using slow senescing stay-green genotypes is a possible means to improve yield in wheat. Ethyl methanesulfonate (EMS) mutated wheat lines (Triticum aestivum L. cv. Paragon) were screened for fast and slow canopy senescence to investigate the impact on yield and nitrogen partitioning. Stay-green and fast-senescing lines with similar anthesis dates were characterised in detail. Delayed senescence was only apparent at higher nitrogen supply with low nitrogen supply enhancing the rate of senescence in all lines. In the stay-green line 3 (SG3), on a whole plant basis, tiller and seed number increased whilst thousand grain weight (TGW) decreased; although a greater N uptake was observed in the main tiller, yield was not affected. In fast-senescing line 2 (FS2), yield decreased, principally as a result of decreased TGW. Analysis of N-partitioning in the main stem indicated that although the slow-senescing line had lower biomass and consequently less nitrogen in all plant parts, the proportion of biomass and nitrogen in the flag leaf was greater at anthesis compared to the other lines; this contributed to the grain N and yield of the slow-senescing line at maturity in both the main tiller and in the whole plant. A field trial confirmed senescence patterns of the two lines, and the negative impact on yield for FS2 and a positive impact for SG3 at low N only. The lack of increased yield in the slow-senescing line was likely due to decreased biomass and additionally a possible sink limitation.

A Major Locus on Wheat Chromosome 7B Associated With Late-Maturity α-Amylase Encodes a Putative ent-Copalyl Diphosphate Synthase.

Many wheat varieties have the potential to develop unacceptably high levels of α-amylase in the grains if exposed to a cool temperature shock or simply cool temperature during the early to middle stages of grain filling. This phenomenon is referred to as late maturity α-amylase (LMA). The enzyme persists in the grain until harvest and may result in wheat with a low Falling Number that does not meet receival and export specifications. Resistance to LMA is therefore a valuable target for wheat breeders and wheat industries in general. Genetic evidence implicating a locus on the long arm of chromosome 7B in variation in LMA phenotype was confirmed in this investigation. Through intensive fine-mapping an ent-copalyl diphosphate synthase (CPS), hitherto named LMA-1, was identified as the likely candidate gene associated with variation in LMA phenotype. Single Nucleotide Polymorphisms (SNPs) within the LMA-1 coding sequence of Chinese Spring, Maringa and Halberd result in either prematurely terminated or functionally altered proteins that are associated with useful levels of resistance to LMA. LMA-1 transcripts detected in de-embryonated grain tissue from around 15 days after anthesis, several days before the synthesis of α-amylase, were low in the resistant varieties Chinese Spring and Maringa compared with LMA susceptible genotype Spica. This was associated with a dramatic reduction in the concentrations of intermediates in the gibberellin biosynthesis pathway such as GA19, evidence that LMA-1 was functioning as CPS in the gibberellin biosynthesis pathway. A survey of a large collection of Australian and international wheat varieties distinguished 9 major haplotypes at the LMA-1 locus. Generally, within classes, there was notable variation for LMA phenotype and evidence for genotypes whose resistance is presumed to be due to genetic loci located elsewhere on the wheat genome. Further investigation is required to characterize the sequence of steps between LMA-1 and α-amylase synthesis as well as to gain a better understanding of the role and potential impact of other genetic loci. Diagnostic markers for sources of resistance and SNP variation reported in this study should assist breeders to deploy resistance associated with LMA-1 variants in breeding programs.

Overgrowth (Della) mutants of wheat: development, growth and yield of intragenic suppressors of the Rht-B1c dwarfing gene.

A suppressor screen using the dwarf Rht-B1c Della mutant of wheat (Triticum aestivum L.) led to the isolation of overgrowth mutants, which retained the original dwarfing gene but grew at a faster rate because of a new mutation elsewhere in that gene. Forty-six alleles were identified, which included amino acid substitutions, premature stop codons, and splice site alterations. The sites of amino acid substitution were primarily localised around conserved motifs in the DELLA protein, and these mutants showed a wide range in their extent of growth recovery (dwarf, semidwarf, tall). Detailed growth comparisons were made on a wide height range of backcrossed overgrowth alleles, comparing stem and spike growth, leaf size, tillering, phenological development, coleoptile length, grain dormancy and grain yield. There were large and reproducible differences between alleles for some traits, whereas others were largely unaffected or varied with growth conditions. Some of the overgrowth alleles offer promise as alternatives to the Rht-B1b and Rht-D1b dwarfing genes, allowing a wider range of height control, improved grain dormancy and equivalent grain yield. The collection of mutants will also be valuable as a resource to study the effect of height on different physiological or agronomic traits, and in elucidating DELLA protein function.