A Heterozygous Genotype-Dependent Branched-Spike Wheat and the Potential Genetic Mechanism Revealed by Transcriptome Sequencing

Wheat (Triticum aestivum L.) spike architecture is an important trait associated with spike development and grain yield. Here, we report a naturally occurring wheat mutant with branched spikelets (BSL) from its wild-type YD-16, which has a normal spike trait and confers a moderate level of resistance to wheat Fusarium head blight (FHB). The lateral meristems positioned at the basal parts of the rachis node of the BSL mutant develop into ramified spikelets characterized as multiple spikelets. The BSL mutant shows three to four-day longer growth period but less 1000-grain weight than the wild type, and it becomes highly susceptible to FHB infection, indicating that the locus controlling the BSL trait may have undergone an intensively artificial and/or natural selection in modern breeding process. The self-pollinated descendants of the lines with the BSL trait consistently segregated with an equal ratio of branched and normal spikelets (NSL) wheat, and homozygotes with the BSL trait could not be achieved even after nine cycles of self-pollination. Distinct segregation patterns both from the self-pollinated progenies of the BSL plants and from the reciprocal crosses between the BSL plants with their sister NSL plants suggested that gametophytic male sterility was probably associated with the heterozygosity for the BSL trait. Transcriptome sequencing of the RNA bulks contrasting in the two types of spike trait at the heading stage indicated that the genes on chromosome 2DS may be critical for the BSL trait formation since 329 out of 2540 differentially expressed genes (DEGs) were located on that chromosome, and most of them were down-regulated. Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis showed that carbohydrate metabolism may be involved in the BSL trait expression. This work provides valuable clues into understanding development and domestication of wheat spike as well as the association of the BSL trait with FHB susceptibility.

Histology and RNA Sequencing Provide Insights Into Fusarium Head Blight Resistance in AAC Tenacious

Fusarium head blight (FHB) is a serious fungal disease affecting wheat and other cereals worldwide. This fungus causes severe yield and quality losses from a reduction in grain quality and contamination of grain with mycotoxins. Intensive breeding efforts led to the release of AAC Tenacious, which was the first spring wheat cultivar registered in Canada with a resistant (R) rating to FHB. To elucidate the physiological mechanisms of resistance, we performed histological and transcriptomic analyses of AAC Tenacious and a susceptible control Roblin after inoculation with Fusarium graminearum (Fg). The spikelet and rachis of infected wheat spikes were hand sectioned and monitored by confocal and fluorescent microscopy. Visible hyphae were observed within the inoculated spikelets for AAC Tenacious; however, the infection was largely restricted to the point of inoculation (POI), whereas the adjacent florets in Roblin were heavily infected. Significant cell wall thickening within the rachis node below the POI was evident in AAC Tenacious compared to Roblin in response to Fg inoculation. Rachis node and rachilla tissues from the POI and the rachis node below the POI were collected at 5 days post inoculation for RNAseq. Significant changes in gene expression were detected in both cultivars in response to infection. The rachis node below the POI in AAC Tenacious had fewer differentially expressed genes (DEGs) when compared to the uninoculated control, likely due to its increased disease resistance. Analysis of DEGs in Roblin and AAC Tenacious revealed the activation of genes and pathways in response to infection, including those putatively involved in cell wall modification and defense response.

The unique disarticulation layer formed in the rachis of Aegilops longissima probably results from the spatial co-expression of Btr1 and Btr2

Background and Aims The brittle rachis trait is a feature of many wild grasses, particularly within the tribe Triticeae. Wild Hordeum and Triticum species form a disarticulation layer above the rachis node, resulting in the production of wedge-type dispersal units. In Aegilops longissima, only one or two of the nodes in the central portion of its rachis are brittle. In Triticeae species, the formation of a disarticulation layer above the rachis node requires the co-transcription of the two dominant and complementary genes Btr1 and Btr2. This study aims to establish whether homologues of Btr1 and/or Btr2 underlie the unusual brittle rachis phenotype observed in Ae. longissima. Methods Scanning electron microscopy was used to examine the disarticulation surfaces. Quantitative RT-PCR and RNA in situ hybridization experiments were used to identify gene expression in the immature inflorescence. Key Results Analysis based on scanning electron microscopy was able to demonstrate that the disarticulation surfaces formed in the Ae. longissima rachis are morphologically indistinguishable from those formed in the rachises of wild Hordeum and Triticum species. RNA in situ hybridization showed that in the immature Ae. longissima inflorescence, the intensity of Btr1 transcription varied from high at the rachis base to low at its apex, while that of Btr2 was limited to the nodes in the central to distal portion of the rachis. Conclusions The disarticulation pattern shown by Ae. longissima results from the limitation of Btr1 and Btr2 co-expression to nodes lying in the centre of the rachis.

Inferring vascular architecture of the wheat spikelet based on resource allocation in the branched headt (bht-A1) near isogenic lines

Substantial genetic and physiological efforts were made to understand the causal factors of floral abortion and grain filling problem in wheat. However, the vascular architecture during wheat spikelet development is surprisingly under-researched. We used the branched headt near-isogenic lines, FL-bht-A1-NILs, to visualise the dynamics of spikelet fertility and dry matter accumulation in spikelets sharing the same rachis node (henceforth Primary Spikelet, PSt, and Secondary Spikelet, SSt). The experiment was conducted after grouping FL-bht-A1-NILs into two groups, where tillers were consistently removed from one group. Our results show differential spikelet fertility and dry matter accumulation between the PSt and SSt, but also showed a concomitant improvement after de-tillering. This suggests a tight regulation of assimilate supply and dry matter accumulation in wheat spikelets. Since PSt and SSt share the same rachis node, the main vascular bundle in the rachis/rachilla is expected to bifurcate to connect each spikelet/floret to the vascular system. We postulate that the vascular structure in the wheat spikelet might even follow Murray’s law, where the wide conduits assigned at the base of the spikelet feed the narrower conduits of the distal florets. We discuss our results based on the two modalities of the vascular network systems in plants.

Developing Kernel and Rachis Node Induce the Trichothecene Pathway of Fusarium graminearum During Wheat Head Infection

The fungal pathogen Fusarium graminearum is the most common agent of Fusarium head blight (FHB) in small grain cereals and cob rot of maize. The threat posed by this fungus is due to a decrease in yield and, additionally, mycotoxin contamination of the harvested cereals. Among the mycotoxins, trichothecenes influence virulence of F. graminearum in a highly complex manner that is strongly host- as well as chemotype-specific. The factors inducing mycotoxin production during plant infection are still unknown. To evaluate the induction of the trichothecene pathway, the green fluorescence protein (GFP) gene was fused to the promoter of the TRI5 gene coding for the trichodiene synthase and integrated into the genome by homologous integration. The resulting mutant contains a fully functional TRI5 gene ensuring virulence on wheat and exhibits GFP driven by the endogenous TRI5 promoter. We are now able to monitor the induction of trichothecenes under real-time conditions. To localize the fungus in the plant tissue, the dsRed gene was integrated under constitutive control of the glycerol-3-phosphate dehydrogenase (gpdA) promoter. We are now able to show that, first, induction of GFP as well as trichothecene production in the reporter strain reflects TRI5 induction and trichothecene production in the wild type; second, expression of TRI5 is inducible during growth in culture; and, third, trichothecene production is not uniformly induced during the onset of infection but is tissue specific during fungal infection of wheat.

Intraspike yield distribution of diverse-tillering spring wheats: Effects of competition

Three spring wheat (Triticum aestivum L.) genotypes (Neepawa, Siete Cerros and M1417) were subjected to four levels of tiller removal (MS remaining, MS + T1 remaining, MS + T1 + T2 remaining, control) at three plant densities (two, four and eight plants per 15-cm-diameter pot) in a controlled environment to assess the effects of interplant and intraplant competition on the intraspike yield distribution of genotypes with diverse tillering habits. Intraspike yield distribution traits varied significantly (P = 0.01) among genotypes and levels of plant competition. Genotypes did not respond uniformly to interplant or intraplant competition. When interplant competition was increased, the oligoculm genotype, M1417, exhibited a greater reduction in maximum kernels per spikelet (13%) and spikelet yield (36%) than either Siete Cerros (9, 17%) or Neepawa (6, 15%). Similarly, with increasing intraplant competition, M1417 responded by reducing maximum kernels per spikelet (6%) and spikelet yield (9%) whereas Siete Cerros and Neepawa exhibited increases for both traits (3 – 15%). The low-tillering genotypes tended to exhibit a greater sensitivity to competition for maximum spikelet yield or kernel number while Neepawa was more responsive for the rachis node at which yield or kernel number was maximized. Strong developmental dominance of the oligoculm habit appears to reduce plasticity and the ability to respond to competition-induced stress. Key words: Spring wheat, intraspike yield, tiller removal, plant competition

Roegneria: its generic limits and justification for its recognition

We document diagnostic characteristics of Roegneria by which it differs from genera with which it has often been confused. Characteristics of Roegneria are the absence of underground runners; slightly curved and elongated rachis with long internodes, each rachis node bearing one spikelet only and often adhering to the internode, and inflorescence appearing more or less one sided because of the disposition of the spikelets; spikelet functionally disarticulating below the glumes; glume symmetric with nerves tapering in the apex; lemma with nerves tapering in the apex; and palea generally shorter than the lemma, and if equal to the lemma then the tip obtuse, truncate, or retuse. Genera difficult to separate from Roegneria are Agropyron, Elymus, and Elytrigia; these share one or more characteristics but none have them in the same combination. We also stress the merit of recognizing Roegneria for germ-plasm utilization. Key words: Roegneria, Triticeae, generic limits, Elymus, Agropyron, Elytrigia.