Rapid gene expression change in a novel synthesized allopolyploid population of cultivated peanut×Arachis doigoi cross by cDNA-SCoT and HFO-TAG technique

AIIopolyploidy has played an important role in plant evolution and heterosis. Recent studies indicate that the process of wide hybridization and （or） polyploidization may induce rapid and extensive genetic and epigenetic changes in some plant species. To better understand the allopolyploidy evolutionism and the genetic mechanism of Arachis interspecific hybridization, this study was conducted to monitor the gene expression variation by cDNA start codon targeted polymorphism （cDNA-SCoT） and cDNA high-frequency oligonucleotide-targeting active gene （cDNA-HFO-TAG） techniques, from the hybrids （F1） and newly synthesized allopolyploid generations （S0-$3） between tetraploid cultivated peanut Zhongkaihua 4 with diploid wild one Arachis doigoi. Rapid and considerable gene expression variations began as early as in the FI hybrid or immediately after chromosome doubling. Three types of gene expression changes were observed, including complete silence （gene from progenitors was not expressed in all progenies）, incomplete silence （gene expressed only in some progenies） and new genes activation. Those silent genes mainly involved in RNA transcription, metabolism, disease resistance, signal transduction and unknown functions. The activated genes with known function were almost retroelements by cDNA-SCoT technique and all metabolisms by cDNA-HFO-TAG. These findings indicated that interspecific hybridization and ploidy change affected gene expression via genetic and epigenetic alterations immediately upon allopolyploid formation, and some obtained transcripts derived fragments （TDFs） probably could be used in the research of molecular mechanism of Arachis allopolyploidization which contribute to thwe genetic diploidization of newly formed allopolyploids. Our research is valuable for understanding of peanut evolution and improving the utilization of putative and beneficial genes from the wild peanut.

Diploid species phylogeny and evolutionary reticulation indicate early radiation of Ephedra in the Tethys coast

Reconstructing a robust species phylogeny and disentangling the evolutionary and biogeographic history of the gymnosperm genus Ephedra,which has a large genome and rich polyploids,remain a big challenge.Here we reconstructed a transcriptome-based phylogeny of 19 diploid Ephedra species,and explored evolutionary reticulations in this genus represented by 50 diploid and polyploid species,using four low-copy nuclear and nine plastid genes.The diploid species phylogeny indicates that the Mediterranean species diverged first,and the remaining species split into three clades,including the American species(Clade A),E.rhytidosperma,and all other Asian species(Clade B).The single-gene trees placed E.rhytidosperma sister to Clade A,Clade B,or Clades A+B in similar proportions,suggesting that radiation and gene flow likely occurred in the early evolution of Ephedra.In addition,reticulate evolution occurred not only among the deep nodes,but also in the recently evolved South American species,which further caused difficulty in phylogenetic reconstruction.Moreover,we found that allopolyploid speciation was pervasive in Ephedra.Our study also suggests that Ephedra very likely originated in the Tethys coast during the late Cretaceous,and the South American Ephedra species have a single origin by dispersal from Mexico or North America.

Making the Bread： Insights from Newly Synthesized AIIohexaploid Wheat

Bread wheat （or common wheat, Triticum aestivum） is an allohexaploid （AABBDD, 2n = 6x = 42） that arose by hybridization between a cultivated tetraploid wheat T. turgidum （AABB, 2n = 4x = 28） and the wild goatgrass Aegilops tauschfi （DD, 2n = 2x = 14）. Polyploidization provided niches for rigorous genome modification at cytogenetic, genetic, and epigenetic levels, rendering a broader spread than its progenitors. This review summarizes the latest advances in understanding gene regulation mechanisms in newly synthesized allo- hexaploid wheat and possible correlation with polyploid growth vigor and adaptation. Cytogenetic studies reveal persistent association of whole-chromosome aneuploidy with nascent allopolyploids, in contrast to the genetic stability in common wheat. Transcriptome analysis of the euploid wheat shows that small RNAs are driving forces for homoeo-allele expression regulation via genetic and epigenetic mechanisms. The ensuing non-additively expressed genes and those with expression level dominance to the respective pro- genitor may play distinct functions in growth vigor and adaptation in nascent allohexaploid wheat. Further genetic diploidization of allohexaploid wheat is not random. Regional asymmetrical gene distribution, rather than subgenome dominance, is observed in both synthetic and natural allohexaploid wheats. The combinatorial effects of diverged genomes, subsequent selection of specific gene categories, and subgenome-specific traits are essential for the successful establishment of common wheat.

Identification of Flowering Regulatory Genes in Allopolyploid Brassica juncea

Brassica juncea is an allopolyploid originating from the interspecific hybridization between Brassica rapa and Brassica nigra, which is of multiple usage as a vegetable, oilseed and condiment worldwide. Both vernalization and non-vernalization under long-day photoperiod can promote floral transition in B. juncea suggesting merged flowering pathways of its ancestors and better environmental adaptability. We identified genomewide flowering regulatory genes in B. juncea, which include 84 and 79 genes from A and B sub-genomes, respectively. Ka/Ks analysis revealed a purification effect on both photoperiod and vernalization flowering regulation pathways during evolution. Expression profile of those genes during long-day and vernalization treatments suggested Bju ACO4, Bju AFT1, Bju BFT4, Bju ASOC1 and Bju ASOC4 may be the major functional copies of B. juncea flowering regulation. Further functional studies about Bju COs showed three days delayed flowering time in Bju ACO4 or Bju BCO3 silenced plants. Increased transcription of all BjuFLCs in Bju ACO4 or Bju BCO3 silenced plants suggested interactions between photoperiod and vernalization pathways governing flowering time. Our findings provided flowering regulating networks in allopolyploid B. juncea.

Generality and characteristics of genetic and epigenetic changes in newly synthesized allotetraploid wheat lines

Previous studies have shown rapid and extensive genomic However, these studies are based on either a few pre-selected instability associated with early stages of allopolyploidization in wheat. genomic loci or genome-wide analysis of a single plant individual for a given cross combination, thus making the extent and generality of the changes uncertain. To further study the generality and characteristics of allopolyploidization-induced genomic instability in wheat, we investigated genetic and epigenetic changes from a genome-wide perspective （by using the AFLP and MSAP markers） in four sets of newly synthesized allotetraploid wheat lines with various genome constitutions, each containing three randomly chosen individual plants at the same generation. We document that although general chromosomal stability was characteristic of all four sets of allotetraploid wheat lines, genetic and epigenetic changes at the molecular level occurred in all these plants, with both kinds of changes classifiable into two distinct categories, i.e., stochastic and directed. The abundant type of genetic change is loss of parental bands while the prevalent cytosine methylation pattern alteration is hypermethylation at the CHG sites. Our results have extended previous studies regarding allopolyploidization-induced genomic dynamics in wheat by demonstrafing the generality of both genetic and epigenetic changes associated with multiple nascent allotetraploid wheat lines, and providing novel insights into the characteristics of the two kinds of induced genomic instabilities.

Maintenance of Species Differences in Closely Related Tetraploid Parasitic Euphrasia (Orobanchaceae) on an Isolated Island

Polyploidy is pervasive in angiosperm evolution and plays important roles in adaptation and speciation.However,polyploid groups are understudied due to complex sequence homology,challenging genome assembly,and taxonomic complexity.Here,we study adaptive divergence in taxonomically complex eyebrights(Euphrasia),where recent divergence,phenotypic plasticity,and hybridization blur species boundaries.We focus on three closely related tetraploid species with contrasting ecological preferences that are sympatric on Fair Isle,a small isolated island in the British Isles.Using a common garden experiment,we show a genetic component to the morphological differences present between these species.Using wholegenome sequencing and a novel k-mer approach we call“Tetmer”,we demonstrate that the species are of allopolyploid origin,with a sub-genome divergence of approximately 5%.Using2 million SNPs,we show sub-genome homology across species,with a very low sequence divergence characteristic of recent speciation.This genetic variation is broadly structured by species,with clear divergence of Fair Isle heathland Euphrasia micrantha,while grassland Euphrasia arctica and coastal Euphrasia foulaensis are more closely related.Overall,we show that tetraploid Euphrasia is a system of allopolyploids of postglacial species divergence,where adaptation to novel environments may be conferred by old variants rearranged into new genetic lineages.