The mRNA decapping machinery targets LBD3/ASL9 to mediate apical hook and lateral root development.

Multicellular organisms perceive and transduce multiple cues to optimize development. Key transcription factors drive developmental changes, but RNA processing also contributes to tissue development. Here, we report that multiple decapping deficient mutants share developmental defects in apical hook, primary and lateral root growth. More specifically, LATERAL ORGAN BOUNDARIES DOMAIN 3 (LBD3)/ASYMMETRIC LEAVES 2-LIKE 9 (ASL9) transcripts accumulate in decapping deficient plants and can be found in complexes with decapping components. Accumulation of ASL9 inhibits apical hook and lateral root formation. Interestingly, exogenous auxin application restores lateral roots formation in both ASL9 over-expressors and mRNA decay-deficient mutants. Likewise, mutations in the cytokinin transcription factors type-B ARABIDOPSIS RESPONSE REGULATORS (B-ARRs) ARR10 and ARR12 restore the developmental defects caused by over-accumulation of capped ASL9 transcript upon ASL9 overexpression. Most importantly, loss-of-function of asl9 partially restores apical hook and lateral root formation in both dcp5-1 and pat triple decapping deficient mutants. Thus, the mRNA decay machinery directly targets ASL9 transcripts for decay, possibly to interfere with cytokinin/auxin responses, during development.

Wheat NAM genes regulate the majority of early monocarpic senescence transcriptional changes including nitrogen remobilization genes.

Senescence enables the remobilization of nitrogen and micronutrients from vegetative tissues of wheat (Triticum aestivum L.) into the grain. Understanding the molecular players in this process will enable the breeding of wheat lines with tailored grain nutrient content. The NAC transcription factor NAM-B1 is associated with earlier senescence and higher levels of grain protein, iron, and zinc contents due to increased nutrient remobilization. To investigate how related NAM genes control nitrogen remobilization at the molecular level, we carried out a comparative transcriptomic study using flag leaves at 7 time points (3, 7, 10, 13, 15, 19, and 26 days after anthesis) in wild type and NAM RNA interference lines with reduced NAM gene expression. Approximately 2.5 times more genes were differentially expressed in wild type than NAM RNA interference plants during this early senescence time course (6,508 vs 2,605 genes). In both genotypes, differentially expressed genes were enriched for gene ontology terms related to photosynthesis, hormones, amino acid transport, and nitrogen metabolism. However, nitrogen metabolism genes including glutamine synthetase (GS1 and GS2), glutamate decarboxylase (GAD), glutamate dehydrogenase (GDH), and asparagine synthetase (ASN1) showed stronger or earlier differential expression in wild-type than in NAM RNA interference plants, consistent with higher nitrogen remobilization. The use of time course data identified the dynamics of NAM-regulated and NAM-independent gene expression changes during senescence and provides an entry point to functionally characterize the pathways regulating senescence and nutrient remobilization in wheat.

The meiotic cohesin subunit REC8 contributes to multigenic adaptive evolution of autopolyploid meiosis in Arabidopsis arenosa.

Genome duplication, which leads to polyploidy, poses challenges to the meiotic segregation of the now-multiple homologous chromosome copies. Genome scan data showed previously that adaptation to polyploid meiosis in autotetraploid Arabidopsis arenosa is likely multigenic, involving genes encoding interacting proteins. But what does this really mean? Functional follow-up studies to genome scans for multigenic traits remain rare in most systems, and thus many mysteries remain about the "functional architecture" of polygenic adaptations. Do different genes all contribute subtle and additive progression towards a fitness optimum, or are there more complex interactions? We previously showed that derived alleles of genes encoding two interacting meiotic axis proteins (ASY1 and ASY3) have additive functional consequences for meiotic adaptation. Here we study derived versus ancestral alleles of the meiotic cohesin subunit REC8, which has roles in chromatin condensation, recruiting the axes, and other critical functions in meiosis. We use genetic and cytological approaches to assess the functional effects of REC8 diploid versus tetraploid alleles, as well as their interaction with ancestral versus derived alleles of ASY1 and ASY3. We show that homozygotes for derived (tetraploid) REC8 alleles have significantly fewer unpaired univalents, a common problem in neotetraploids. Interactions with ASY1 and ASY3 are complex, with the genes in some cases affecting distinct traits, and additive or even antagonistic effects on others. These findings suggest that the road to meiotic adaptation in A. arenosa was perhaps neither straight nor smooth.

Comparative Mapping and Targeted-Capture Sequencing of the Gametocidal Loci in Aegilops sharonensis.

Gametocidal (Gc) chromosomes or elements in species such as Eig are preferentially transmitted to the next generation through both the male and female gametes when introduced into wheat ( L.). Furthermore, any genes, such as genes that control agronomically important traits, showing complete linkage with Gc elements, are also transmitted preferentially to the next generation without the need for selection. The mechanism for the preferential transmission of the Gc elements appears to occur by the induction of extensive chromosome damage in any gametes that lack the Gc chromosome in question. Previous studies on the mechanism of the Gc action in indicates that at least two linked elements are involved. The first, the element, induces chromosome breakage in gametes, which have lost the Gc elements while the second, the element, prevents the chromosome breakage action of the breaker element in gametes which carry the Gc elements. In this study, we have used comparative genomic studies to map 54 single nucleotide polymorphism (SNP) markers in an 4S introgression segment in wheat and have also identified 18 candidate genes in for the breaker element through targeted sequencing of this 4S introgression segment. This valuable genomic resource will aide in further mapping the Gc locus that could be exploited in wheat breeding to produce new, superior varieties of wheat.

Mapping the 'breaker' element of the gametocidal locus proximal to a block of sub-telomeric heterochromatin on the long arm of chromosome 4S(sh) of Aegilops sharonensis.

KEY MESSAGE: The 'breaker' element ( GcB ) of the gametocidal locus derived from Aegilops sharonensis has been mapped to a region proximal to a block of sub-telomeric heterochromatin on chromosome 4S (sh) L. The production of alien chromosome addition lines allows the transfer of useful genetic variation into elite wheat varieties from related wild species. However, some wild relatives of wheat, particularly those within the Sitopsis section of the genus Aegilops, possess chromosomes that are transmitted preferentially to the offspring when addition lines are generated. Species within the Sitopsis group possess the S genome, and among these species, Aegilops sharonensis (2n = 14, S(sh)S(sh)) carries the S(sh) genome which is closely related to the D genome of hexaploid wheat. Some S genome chromosomes carry gametocidal loci, which induce severe chromosome breakage in gametes lacking the gametocidal chromosome, and hence, result in gamete abortion. The preferential transmission of gametocidal loci could be exploited in wheat breeding, because linking gametocidal loci with important agronomic traits in elite wheat varieties would ensure retention of these traits through successive generations. In this study, we have mapped the breaker element of the gametocidal locus derived from Ae. sharonensis to the region immediately proximal to a block of sub-telomeric heterochromatin on the long arm of chromosome 4S(sh).

Inducing chromosome pairing through premature condensation: analysis of wheat interspecific hybrids.

At the onset of meiosis, chromosomes first decondense and then condense as the process of recognition and intimate pairing occurs between homologous chromosomes. We show here that okadaic acid, a drug known to induce chromosome condensation, can be introduced into wheat interspecific hybrids prior to meiosis to induce chromosome pairing. This pairing occurs in the presence of the Ph1 locus, which usually suppresses pairing of related chromosomes and which we show here delays condensation. Thus the timing of chromosome condensation during the onset of meiosis is an important factor in controlling chromosome pairing.

Development of consistently crossable wheat genotypes for alien wheat gene transfer through fine-mapping of the Kr1 locus.

Breeders can force sexual hybridisation between wheat and related grass species to produce interspecific hybrids containing a dihaploid set of wheat and related chromosomes. This facilitates the introgression of desirable genes into wheat from the secondary gene pool. However, most elite European wheat varieties carry genes that suppress crossability, making the transfer of novel traits from exotic germplasm into elite wheat varieties difficult or impossible. Previous studies have identified at least five crossability loci in wheat. Here, the crossability locus with the largest effect, Kr1 on chromosome arm 5BL, was fine-mapped by developing a series of recombinant substitution lines in which the genome of the normally non-crossable wheat variety 'Hobbit sib' carries a recombinant 5BL chromosome arm containing segments from the crossable variety 'Chinese Spring'. These recombinant lines were scored for their ability to cross with rye over four seasons. Analysis revealed at least two regions on 5BL affecting crossability, including the Kr1 locus. However, the ability to set seed is highly dependent on prevailing environmental conditions. Typically, even crossable wheat lines exhibit little or no seed set when crossed with rye in winter, but show up to 90% seed set from similar crosses made in summer. By recombining different combinations of the two regions affecting crossability, wheat lines that consistently exhibit up to 50% seed set, whether crossed in the UK winter or summer conditions, were generated, thus creating a very important tool for increasing the efficiency of alien wheat transfer programmes.

Detailed dissection of the chromosomal region containing the Ph1 locus in wheat Triticum aestivum: with deletion mutants and expression profiling.

BACKGROUND AND AIMS: Understanding Ph1, a dominant homoeologous chromosome pairing suppressor locus on the long arm of chromosome 5B in wheat Triticum aestivum L., is the core of the investigation in this article. The Ph1 locus restricts chromosome pairing and recombination at meiosis to true homologues. The importance of wheat as a crop and the need to exploit its wild relatives as donors for economically important traits in wheat breeding programmes is the main drive to uncover the mechanism of the Ph1 locus and regulate its activity. METHODS: Following the molecular genetic characterization of the Ph1 locus, five additional deletion mutants covering the region have been identified. In addition, more bacterial artificial chromosomes (BACs) were sequenced and analysed to elucidate the complexity of this locus. A semi-quantitative RT-PCR was used to compare the expression profiles of different genes in the 5B region containing the Ph1 locus with their homoeologues on 5A and 5D. PCR products were cloned and sequenced to identify the gene from which they were derived. KEY RESULTS: Deletion mutants and expression profiling of genes in the region containing the Ph1 locus on 5B has further restricted Ph1 to a cluster of cdk-like genes. Bioinformatic analysis of the cdk-like genes revealed their close homology to the checkpoint kinase Cdk2 from humans. Cdk2 is involved in the initiation of replication and is required in early meiosis. Expression profiling has revealed that the cdk-like gene cluster is unique within the region analysed on 5B in that these genes are transcribed. Deletion of the cdk-like locus on 5B results in activation of transcription of functional cdk-like copies on 5A and 5D. Thus the cdk locus on 5B is dominant to those on 5A and 5D in determining the overall activity, which will be dependent on a complex interplay between transcription from non-functional and functional cdk-like genes. CONCLUSIONS: The Ph1 locus has been defined to a cdk-like gene cluster related to Cdk2 in humans, a master checkpoint gene involved in the initiation of replication and required for early meiosis.