PnTgs1-like expression during reproductive development supports a role for RNA methyltransferases in the aposporous pathway

International audienceBackground:In flowering plants, apomixis (asexual reproduction via seeds) is widely believed to result from failureof key regulators of the sexual female reproductive pathway. In the past few years, both differential display andRNA-seq comparative approaches involving reproductive organs of sexual plants and their apomictic counterparts haveyielded extensive lists of candidate genes. Nevertheless, only a limited number of these genes have been functionallycharacterized, with few clues consequently available for understanding the molecular control of apomixis. We havepreviously identified several cDNA fragments with high similarity to genes involved in RNA biology and with differentialamplification between sexual and apomicticPaspalum notatumplants. Here, we report the characterization of one ofthese candidates, namely, N69 encoding a protein of the S-adenosyl-L-methionine-dependent methyltransferasessuperfamily. The purpose of this work was to extend the N69 cDNA sequence and to characterize its expression atdifferent developmental stages in both sexual and apomictic individuals.Results:Molecular characterization of the N69 cDNA revealed homology with genes encoding proteins similar to yeastand mammalian trimethylguanosine synthase/PRIP-interacting proteins. These proteins play a dual role as ERK2-controlledtranscriptional coactivators and mediators of sn(o)RNA and telomerase RNA cap trimethylation, and participate inmammals and yeast development. The N69-extended sequence was consequently renamedPnTgs1-like.ExpressionofPnTgs1-likeduring reproductive development was significantly higher in floral organs of sexual genotypes comparedwith apomicts. This difference was not detected in vegetative tissues. In addition, expression levels in reproductivetissues of several genotypes were negatively correlated with facultative apomixis rates. Moreover,in situhybridizationobservations revealed thatPnTgs1-likeexpression is relatively higher in ovules of sexual plants throughout development,from premeiosis to maturity. Tissues where differential expression is detected include nucellar cells, the site ofaposporous initials differentiation in apomictic genotypes.Conclusions:Our results indicate thatPnTgs1-like(formerly N69) encodes a trimethylguanosine synthase-likeprotein whose function in mammals and yeast is critical for development, including reproduction. Our findingsalso suggest a pivotal role for this candidate gene in nucellar cell fate, as its diminished expression is correlated withinitiation of the apomictic pathway in plants

The vesicle trafficking regulator PN_SCD1 is demethylated and overexpressed in florets of apomictic Paspalum notatum genotypes

Abstract Apomixis (asexual reproduction through seeds) is considered a deviation of the sexual reproductive pathway leading to the development of clonal progenies genetically identical to the mother plant. Here we used the Methylation-Sensitive Amplification Polymorphism (MSAP) technique to characterize cytosine methylation patterns occurring in florets of sexual and aposporous Paspalum notatum genotypes, in order to identify epigenetically-controlled genes putatively involved in apomixis development. From twelve polymorphic MSAP-derived sequences, one (PN_6.6, later renamed PN_SCD1) was selected due to its relevant annotation and differential representation in apomictic and sexual floral transcriptome libraries. PN_SCD1 encodes the DENN domain/WD repeat-containing protein SCD1, which interacts with RAB GTPases- and/or MAPKs to promote specialized cell division, functions in clathrin-mediated membrane transport and acts as potential substrate receptor of CUL4 E3 ubiquitin ligases. Quantitative RT-PCR and comparative RNAseq analyses of laser microdissected nucellar cells confirmed PN_SCD1 upregulation in florets of apomictic plants and revealed that overexpression takes place just before the onset of apospory initials. Moreover, we found that several SCD1 molecular partners are expressed in P. notatum florets and upregulated in apomictic plants. Our results disclosed a specific vesicle trafficking molecular pathway epigenetically modulated during apomixis

A Portion of the Apomixis Locus of Paspalum Simplex is Microsyntenic with an Unstable Chromosome Segment Highly Conserved Among Poaceae

Abstract The introgression of apomixis in major seed crops, would guarantee self-seeding of superior heterotic seeds over generations. In the grass species Paspalum simplex, apomixis is controlled by a single locus in which recombination is blocked. In the perspective of isolating the genetic determinants of apomixis, we report data on sequencing, in silico mapping and expression analysis of some of the genes contained in two cloned genomic regions of the apomixis locus of P. simplex. In silico mapping allowed us to identify a conserved synteny group homoeologous to the apomixis locus, located on a telomeric position of chromosomes 12, 8, 3 and 4 of rice, Sorghum bicolor, Setaria italica and Brachypodium distachyum, respectively, and on a more centromeric position of maize chromosome 1. Selected genes of the apomixis locus expressed sense and antisense transcripts in reproductively committed cells of sexual and apomictic ovules. Some of the genes considered here expressed apomixis-specific allelic variants which showed partial non-overlapping expression patterns with alleles shared by sexual and apomictic reproductive phenotypes. Our findings open new routes for the isolation of the genetic determinants of apomixis and, in perspective, for its introgression in crop grasses

The MAP3K-Coding QUI-GON JINN (QGJ) gene is essential to the formation of unreduced embryo sacs in Paspalum.

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