Polyploid species have long been thought to be recalcitrant to whole-genome assembly. By combining high-throughput sequencing, recent developments in parallel computing, and genetic mapping, we derive, de novo, a sequence assembly representing 9.1 Gbp of the highly repetitive 16 Gbp genome of hexaploid wheat, Triticum aestivum, and assign 7.1 Gb of this assembly to chromosomal locations. The genome representation and accuracy of our assembly is comparable or even exceeds that of a chromosome-by-chromosome shotgun assembly. Our assembly and mapping strategy uses only short read sequencing technology and is applicable to any species where it is possible to construct a mapping population.

A whole-genome shotgun approach for assembling and anchoring the hexaploid bread wheat genome

Gene expression can be regulated through transcriptional and post-transcriptional mechanisms. Transcription in eukaryotes produces pre-mRNA molecules, which are processed and spliced post-transcriptionally to create translatable mRNAs. More than one mRNA may be produced from a single pre-mRNA by alternative splicing (AS); thus, AS serves to diversify an organism's transcriptome and proteome. Previous studies of gene expression in plants have focused on the role of transcriptional regulation in response to environmental changes. However, recent data suggest that post-transcriptional regulation, especially AS, is necessary for plants to adapt to a changing environment. In this review, we summarize recent advances in our understanding of AS during plant development in response to environmental changes. We suggest that alternative gene splicing is a novel means of regulating the environmental fitness of plants.

Alternative Splicing in Plant Genes: A Means of Regulating the Environmental Fitness of Plants.

Transcript elongation factors (TEFs) are a heterogeneous group of proteins that control the efficiency of transcript elongation of subsets of genes by RNA polymerase II (RNAPII) in the chromatin context. Using reciprocal tagging in combination with affinity purification and mass spectrometry, we demonstrate that in Arabidopsis thaliana, the TEFs SPT4/SPT5, SPT6, FACT, PAF1-C, and TFIIS copurified with each other and with elongating RNAPII, while P-TEFb was not among the interactors. Additionally, NAP1 histone chaperones, ATP-dependent chromatin remodeling factors, and some histone-modifying enzymes including Elongator were repeatedly found associated with TEFs. Analysis of double mutant plants defective in different combinations of TEFs revealed genetic interactions between genes encoding subunits of PAF1-C, FACT, and TFIIS, resulting in synergistic/epistatic effects on plant growth/development. Analysis of subnuclear localization, gene expression, and chromatin association did not provide evidence for an involvement of the TEFs in transcription by RNAPI (or RNAPIII). Proteomics analyses also revealed multiple interactions between the transcript elongation complex and factors involved in mRNA splicing and polyadenylation, including an association of PAF1-C with the polyadenylation factor CstF. Therefore, the RNAPII transcript elongation complex represents a platform for interactions among different TEFs, as well as for coordinating ongoing transcription with mRNA processing.

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The Composition of the Arabidopsis RNA Polymerase II Transcript Elongation Complex Reveals the Interplay Between Elongation and mRNA Processing Factors

N-terminal acetylation (NTA) is a prevalent protein modification in eukaryotes. The majority of proteins are acetylated at their N-terminus in a co-translational manner by ribosome-associated N-terminal acetyltransferases (NATs). However, the recent discovery of Golgi membrane-localized NATs in metazoa, and plastid-localized NATs in plants challenged the dogma of static, co-translational imprinting of the proteome by NTA. Indeed, NTA by the cytosolic NatA is highly dynamic and under hormonal control in plants. Such active control has not been evidenced yet in other eukaryotes and might be an adaptation to the sessile lifestyle of plants forcing them to cope with diverse environmental challenges. The function of NTAs for individual proteins is distinct and yet unpredictable. In yeast and humans, NTA has been shown to affect protein-protein interactions, subcellular localization, folding, aggregation, or degradation of a handful of proteins. In particular, the impact of NTA on the protein turnover is documented by diverse examples in yeast. Consequently, NTA was recently dicovered to be a degradation signal in a distinct branch of the N-end rule pathway, ubiquitin-mediated proteolysis. In this review, we summarize the current knowledge on the NAT machinery in higher plants and discuss the potential function of NTA during biotic and abiotic stresses.

/pdf/n-terminal-acetylation-an-essential-protein-modification-5wy1jky6hc.pdf

N-terminal acetylation: an essential protein modification emerges as an important regulator of stress responses.

RNA synthesis by RNA polymerase II and RNA processing are closely coupled during the transcription cycle of protein-coding genes. This coupling affords opportunities for quality control and regulation of gene expression and the effects can go in both directions. For example, polymerase speed can affect splice site selection and splicing can increase transcription and affect the chromatin landscape. Here we review the many ways that transcription and splicing influence one another, including how splicing "talks back" to transcription. We will also place the connections between transcription and splicing in the context of other RNA processing events that define the exons that will make up the final mRNA. This article is categorized under: RNA Processing > Splicing Mechanisms RNA Processing > Splicing Regulation/Alternative Splicing.

/pdf/transcription-and-splicing-a-two-way-street-xyf9fx0lkq.pdf

Transcription and splicing: A two-way street.

Seed size is a pivotal agronomic trait that links plant sexual reproduction and subsequent seedling establishment, and is affected by the timing of endosperm cellularization following endosperm proliferation after double fertilization. The molecular switch that controls the timing of endosperm cellularization has so far been largely unclear. Here, we report that the Arabidopsis TERMINAL FLOWER1 (TFL1) is a mobile regulator generated in the chalazal endosperm, and moves to the syncytial peripheral endosperm to mediate timely endosperm cellularization and seed size through stabilizing ABSCISIC ACID INSENSITIVE 5. We further show that Ras-related nuclear GTPases interact with TFL1 and regulate its trafficking to the syncytial peripheral endosperm. Our findings reveal TFL1 as an essential molecular switch for regulating endosperm cellularization and seed size. Generation of mobile TFL1 in the chalazal endosperm, which is close to maternal vascular tissues, could provide a hitherto-unknown means to control seed development by mother plants.

Mobile TERMINAL FLOWER1 determines seed size in Arabidopsis.

FLOWERING LOCUS T (FT) in Arabidopsis encodes the florigen that moves from leaves to the shoot apical meristem to induce flowering, and this is partly mediated by FT-INTERACTING PROTEIN 1 (FTIP1). Although FT orthologs have been identified in some flowering plants, their endogenous roles in Orchidaceae, which is one of the largest families of flowering plants, are still largely unknown. In this study, we show that DOFT and DOFTIP1, the orchid orthologs of FT and FTIP1, respectively, play important roles in promoting flowering in the orchid Dendrobium Chao Praya Smile. Expression of DOFT and DOFTIP1 increases in whole plantlets during the transition from vegetative to reproductive development. Both transcripts are present in significant levels in reproductive organs, including inflorescence apices, stems, floral buds, and open flowers. Through successful generation of transgenic orchids, we have revealed that overexpression or down-regulation of DOFT accelerates or delays flowering, respectively, while alteration of DOFT expression also greatly affects pseudobulb formation and flower development. In common with their counterparts in Arabidopsis and rice, DOFTIP1 interacts with DOFT and affects flowering time in orchids. Our results suggest that while DOFT and DOFTIP1 play evolutionarily conserved roles in promoting flowering, DOFT may have evolved with hitherto unknown functions pertaining to the regulation of storage organs and flower development in the Orchidaceae family.

DOFT and DOFTIP1 affect reproductive development in the orchid Dendrobium Chao Praya Smile

Ski-interacting protein (SKIP) is a bifunctional regulator of gene expression that works as a splicing factor as part of the spliceosome and as a transcriptional activator by interacting with EARLY FLOWERING 7 (ELF7). MOS4-Associated Complex 3A (MAC3A) and MAC3B interact physically and genetically with SKIP, mediate the alternative splicing of c. 50% of the expressed genes in the Arabidopsis genome, and are required for the splicing of a similar set of genes to that of SKIP. SKIP interacts physically and genetically with splicing factors and Polymerase-Associated Factor 1 complex (Paf1c) components. However, these splicing factors do not interact either physically or genetically with Paf1c components. The SKIP-spliceosome complex mediates circadian clock function and abiotic stress responses by controlling the alternative splicing of pre-mRNAs encoded by clock- and stress tolerance-related genes. The SKIP-Paf1c complex regulates the floral transition by activating FLOWERING LOCUS C (FLC) transcription. Our data reveal that SKIP regulates floral transition and environmental fitness via its incorporation into two distinct complexes that regulate gene expression transcriptionally and post-transcriptionally, respectively. It will be interesting to discover in future studies whether SKIP is required for integration of environmental fitness and growth by control of the incorporation of SKIP into spliceosome or Paf1c in plants.

https://nph.onlinelibrary.wiley.com/doi/epdf/10.1111/nph.15990

SKIP regulates environmental fitness and floral transition by forming two distinct complexes in Arabidopsis.

Early embryonic development generates precursors of all major cell types in Arabidopsis. Among these precursors, the hypophysis divides asymmetrically to form the progenitors of the quiescent center and columella stem cells. A great deal has been learnt about the mechanisms that control the asymmetric division of the hypophysis and embryogenesis at the transcriptional level; however, no evidence of regulation at the co- or post-translational level has been reported. Here, we show that mutation of the catalytic subunit (Naa10) or auxiliary subunit (Naa15) of NatA, an N-terminal acetyltransferase that catalyzes protein N-terminal acetylation, produces an embryo-lethal phenotype. In addition, Naa10 and Naa15 were found to interact physically in planta. Further analysis revealed that the observed embryonic patterning defects started at the early globular stage and that the asymmetric division of the hypophysis was irregular; thus, no quiescent center progenitor cells were generated in naa10 and naa15 embryos. We further observed that the polar distributions of auxin and its efflux carrier PIN1 were disturbed in naa10 embryos. Our results suggest that NatA is required for asymmetric division of the hypophysis and early embryonic patterning in Arabidopsis, and provides a link between protein N-terminal acetylation and embryogenesis in plants.

Yan Li

Papers

Mobile TERMINAL FLOWER1 determines seed size in Arabidopsis.

DOFT and DOFTIP1 affect reproductive development in the orchid Dendrobium Chao Praya Smile

SKIP regulates environmental fitness and floral transition by forming two distinct complexes in Arabidopsis.

Protein N-terminal acetylation is required for embryogenesis in Arabidopsis

CRISPR/Cas9-Based Genome Editing and its Applications for Functional Genomic Analyses in Plants