Grass Subfamily Pooideae Research Articles

Some unique anatomical scaling relationships among genera in the grass subfamily Pooideae.

Members of the grass family Poaceae have adapted to a wide range of habitats and disturbance regimes globally. The cellular structure and arrangements of leaves can help explain how plants survive in different climates, but these traits are rarely measured in grasses. Most studies are focussed on individual species or distantly related species within Poaceae. While this focus can reveal broad adaptations, it is also likely to overlook subtle adaptations within more closely related groups (subfamilies, tribes). This study, therefore, investigated the scaling relationships between leaf size, vein length area (VLA) and vessel size in five genera within the subfamily Pooideae. The scaling exponent of the relationship between leaf area and VLA was -0.46 (±0.21), which is consistent with previous studies. In Poa and Elymus, however, minor vein number and leaf length were uncorrelated, whereas in Festuca these traits were positively correlated (slope = 0.82±0.8). These findings suggest there are broad-scale and fine-scale variations in leaf hydraulic traits among grasses. Future studies should consider both narrow and broad phylogenetic gradients.

TaWRKY10 transcription factor is a novel jasmonic acid signalling regulator involved in immunity against Septoria tritici blotch disease in wheat

Septoria tritici blotch (STB), caused by the fungus Zymoseptoria tritici, is currently the main threat to worldwide wheat production in temperate climates. Understanding the genetic mechanisms that underpin the Z. tritici–wheat interaction will be crucial for generating new control strategies against STB. Plant hormones are essential regulators of growth and immunity. In particular, jasmonic acid (JA) plays a central role in defence against necrotrophic plant pathogens, but its role in mediating immunity against Z. tritici is largely unknown. Here we identify the transcription factor TaWRKY10 that is taxonomically restricted to the grass subfamily Pooideae as a novel regulator of JA responses in wheat. We demonstrate by using virus‐induced gene silencing that silencing TaWRKY10 leads to increased resistance against Z. tritici with an earlier onset of necrotic symptoms, but with reduced pathogen sporulation. Wheat plants silenced for TaWRKY10 show an up‐regulated JA response. Transcriptional profiling of TaWRKY10 knock‐down plants indicates that it is a negative regulator of the JA receptor TaCOI1 gene expression. Our data indicate that TaWRKY10 down‐regulates JA responses, contributing to the susceptibility of wheat to Z. tritici. We postulate that manipulating TaWRKY10 may provide a strategy to boost STB resistance in wheat.

Phytolith records of flourishing early Holocene Pooideae linked to an 8.2 ka cold event in subtropical China

The grass subfamily Pooideae originated in a temperate niche during the late Cretaceous; it is the largest Poaceae subfamily, consisting of almost 4,000 species, which are distributed worldwide. Pooideae responses to climate changes at different time scales, and different ecological zones are thus important in understanding Poaceae evolutionary processes and their relationship with climate change. In the study described in this article, we reconstructed Pooideae variability during the early Holocene, as inferred by a phytolith sequence from the Lower Yangtze in subtropical China. The phytolith assemblage was marked by three increases in Pooideae phytoliths, dated to ca 8.4–8.0, 7.8–7.6, and 7.4–7.2 ka BP (before present, 1950 AD), with each representing pronounced increases in Pooideae extent and distribution. All these increases were within age ranges that agreed well with the timing of weak Asian Monsoon events, at 8.2, 7.7, and 7.3 ka BP. The first Pooideae flourishing period in subtropical China was the most significant, lasting for approximately four centuries and being characterized by a double peak, which equated with an event at 8.2 ka. This suggested that cold and/or dry conditions—which occurred over a period of several hundred years and were linked to weakening of the Asian monsoon—probably caused Pooideae to flourish in the Lower Yangtze region. Comparison of two diagnostic trapezoid phytolith types—namely wavy and wavy narrow—which showed different changes between ca 8.4 and 8.0 ka BP, suggested that they responded differently to the climate change represented by the 8.2 ka event. Our phytolith records have provided not only new data clarifying the detailed Pooideae response to the 8.2 ka event but also a reliable index for past cold climates in subtropical China.

Increased above-ground resource allocation is a likely precursor for independent evolutionary origins of annuality in the Pooideae grass subfamily.

Semelparous annual plants flower a single time during their 1-yr life cycle, investing much of their energy into rapid reproduction. By contrast, iteroparous perennial plants flower multiple times over several years, and partition their resources between reproduction and persistence. To which extent evolutionary transitions between life-cycle strategies are internally constrained at the developmental, genetic and phylogenetic level is unknown. Here we study the evolution of life-cycle strategies in the grass subfamily Pooideae and test if transitions between them are facilitated by evolutionary precursors. We integrate ecological, life-cycle strategy and growth data in a phylogenetic framework. We investigate if growth traits are candidates for a precursor. Species in certain Pooideae clades are predisposed to evolve annuality from perenniality, potentially due to the shared inheritance of specific evolutionary precursors. Seasonal dry climates, which have been linked to annuality, were only able to select for transitions to annuality when the precursor was present. Allocation of more resources to above-ground rather than below-ground growth is a candidate for the precursor. Our findings support the hypothesis that only certain lineages can respond quickly to changing external conditions by switching their life-cycle strategy, likely due to the presence of evolutionary precursors.

The grass subfamily Pooideae: Cretaceous–Palaeocene origin and climate‐driven Cenozoic diversification

AimFrost is among the most dramatic stresses a plant can experience, and complex physiological adaptations are needed to endure long periods of sub‐zero temperatures. Owing to the need to evolve these complex adaptations, transitioning from tropical to temperate climates is regarded as difficult. Here, we study the transition from tropical to temperate climates in the grass subfamily Pooideae, which dominates cool temperate, continental and Arctic regions. We produce a dated phylogeny and investigate the role of climate cooling in diversification.LocationGlobal, temperate regions.Time periodCretaceous–Cenozoic.Major taxaPooideae.MethodsUsing newly available fossils and methods, we dated a comprehensive Pooideae phylogeny and tested for the impact of palaeoclimates on diversification rates. Using ancestral state reconstruction, we investigated whether Pooideae ancestors experienced frost and winter. To locate the ancestral distribution area of Pooideae, we performed biogeographical analyses.ResultsWe estimated a Late Cretaceous/early Palaeocene origin of the Pooideae (61–77 Ma), with all major clades already having diversified at the Eocene–Oligocene climate cooling (34 Ma). Climate cooling was a probable driving force of Pooideae diversification. Pooideae probably evolved in a temperate niche experiencing frost, but not long winters.Main conclusionPooideae probably originated in a temperate niche and experienced cold temperatures and frost long before expansion of temperate biomes after the Eocene–Oligocene transition. This suggests that the Pooideae ancestor had adaptations to temperate climate and that certain responses to low‐temperature stress are shared in extant Pooideae grasses. Throughout the Cenozoic, falling temperatures and expansion of temperate biomes were associated with an increase in diversification. However, complex mechanisms for enduring strongly seasonal climate with long, cold winters most probably evolved independently in daughter lineages. Our findings provide insight into how adaptations to historical changes in chill and frost exposure influence the distribution of plant diversity today.

Evolution of Cold Acclimation and Its Role in Niche Transition in the Temperate Grass Subfamily Pooideae.

The grass subfamily Pooideae dominates the grass floras in cold temperate regions and has evolved complex physiological adaptations to cope with extreme environmental conditions like frost, winter, and seasonality. One such adaptation is cold acclimation, wherein plants increase their frost tolerance in response to gradually falling temperatures and shorter days in the autumn. However, understanding how complex traits like cold acclimation evolve remains a major challenge in evolutionary biology. Here, we investigated the evolution of cold acclimation in Pooideae and found that a phylogenetically diverse set of Pooideae species displayed cold acclimation capacity. However, comparing differential gene expression after cold treatment in transcriptomes of five phylogenetically diverse species revealed widespread species-specific responses of genes with conserved sequences. Furthermore, we studied the correlation between gene family size and number of cold-responsive genes as well as between selection pressure on coding sequences of genes and their cold responsiveness. We saw evidence of protein-coding and regulatory sequence evolution as well as the origin of novel genes and functions contributing toward evolution of a cold response in Pooideae. Our results reflect that selection pressure resulting from global cooling must have acted on already diverged lineages. Nevertheless, conservation of cold-induced gene expression of certain genes indicates that the Pooideae ancestor may have possessed some molecular machinery to mitigate cold stress. Evolution of adaptations to seasonally cold climates is regarded as particularly difficult. How Pooideae evolved to transition from tropical to temperate biomes sheds light on how complex traits evolve in the light of climate changes.

Variability of Pyrrolizidine Alkaloid Occurrence in Species of the Grass Subfamily Pooideae (Poaceae).

Pyrrolizidine alkaloids (PAs) are a class of secondary metabolites found in various unrelated angiosperm lineages including cool-season grasses (Poaceae, subfamily Pooideae). Thesinine conjugates, saturated forms of PA that are regarded as non-toxic, have been described to occur in the two grass species Lolium perenne and Festuca arundinacea (Poaceae, subfamily Pooideae). In a wider screen, we tested various species of the Pooideae lineage, grown under controlled conditions, for their ability to produce thesinine conjugates or related structures. Using an LC-MS based targeted metabolomics approach we were able to show that PA biosynthesis in grasses is limited to a group of very closely related Pooideae species that produce a limited diversity of PA structures. High variability in PA levels was observed even between individuals of the same species. These individual accumulation patterns are discussed with respect to a possible function and evolution of this type of alkaloid.

Evolution of the miR5200-FLOWERING LOCUS T flowering time regulon in the temperate grass subfamily Pooideae

Flowering time is a carefully regulated trait controlled primarily through the action of the central genetic regulator, FLOWERING LOCUS T (FT). Recently it was demonstrated that a microRNA, miR5200, targets the end of the second exon of FT under short-day photoperiods in the grass subfamily Pooideae, thus preventing FT transcripts from reaching threshold levels under non-inductive conditions. Pooideae are an interesting group in that they rapidly diversified from the tropics into the northern temperate region during a major global cooling event spanning the Eocene-Oligocene transition. We hypothesize that miR5200 photoperiod-sensitive regulation of Pooideae flowering time networks assisted their transition into northern seasonal environments. Here, we test predictions derived from this hypothesis that miR5200, originally found in bread wheat and later identified in Brachypodium distachyon, (1) was present in the genome of the Pooideae common ancestor, (2) is transcriptionally regulated by photoperiod, and (3) is negatively correlated with FT transcript abundance, indicative of miR5200 regulating FT. Our results demonstrate that miR5200 did evolve at or around the base of Pooideae, but only acquired photoperiod-regulated transcription within the Brachypodium lineage. Based on expression profiles and previous data, we posit that the progenitor of miR5200 was co-regulated with FT by an unknown mechanism.

TaFROG Encodes a Pooideae Orphan Protein That Interacts with SnRK1 and Enhances Resistance to the Mycotoxigenic Fungus Fusarium graminearum.

All genomes encode taxonomically restricted orphan genes, and the vast majority are of unknown function. There is growing evidence that such genes play an important role in the environmental adaptation of taxa. We report the functional characterization of an orphan gene (Triticum aestivum Fusarium Resistance Orphan Gene [TaFROG]) as a component of resistance to the globally important wheat (T. aestivum) disease, Fusarium head blight. TaFROG is taxonomically restricted to the grass subfamily Pooideae. Gene expression studies showed that it is a component of the early wheat response to the mycotoxin deoxynivalenol (DON), which is a virulence factor produced by the causal fungal agent of Fusarium head blight, Fusarium graminearum. The temporal induction of TaFROG by F. graminearum in wheat spikelets correlated with the activation of the defense Triticum aestivum Pathogenesis-Related-1 (TaPR1) gene. But unlike TaPR1, TaFROG induction by F. graminearum was toxin dependent, as determined via comparative analysis of the effects of wild-type fungus and a DON minus mutant derivative. Using virus-induced gene silencing and overexpressing transgenic wheat lines, we present evidence that TaFROG contributes to host resistance to both DON and F. graminearum. TaFROG is an intrinsically disordered protein, and it localized to the nucleus. A wheat alpha subunit of the Sucrose Non-Fermenting1-Related Kinase1 was identified as a TaFROG-interacting protein based on a yeast two-hybrid study. In planta bimolecular fluorescence complementation assays confirmed the interaction. Thus, we conclude that TaFROG encodes a new Sucrose Non-Fermenting1-Related Kinase1-interacting protein and enhances biotic stress resistance.

A multi-locus analysis of phylogenetic relationships within grass subfamily Pooideae (Poaceae) inferred from sequences of nuclear single copy gene regions compared with plastid DNA

To investigate phylogenetic relationships within the grass subfamily Pooideae we studied about 50 taxa covering all recognized tribes, using one plastid DNA (cpDNA) marker (matK gene–3′trnK exon) and for the first time four nuclear single copy gene loci. DNA sequence information from two parts of the nuclear genes topoisomerase 6 (Topo6) spanning the exons 8–13 and 17–19, the exons 9–13 encoding plastid acetyl-CoA-carboxylase (Acc1) and the partial exon 1 of phytochrome B (PhyB) were generated. Individual and nuclear combined data were evaluated using maximum parsimony, maximum likelihood and Bayesian methods. All of the phylogenetic results show Brachyelytrum and the tribe Nardeae as earliest diverging lineages within the subfamily. The ‘core’ Pooideae (Hordeeae and the Aveneae/Poeae tribe complex) are also strongly supported, as well as the monophyly of the tribes Brachypodieae, Meliceae and Stipeae (except PhyB). The beak grass tribe Diarrheneae and the tribe Duthieeae are not monophyletic in some of the analyses. However, the combined nuclear DNA (nDNA) tree yields the highest resolution and the best delimitation of the tribes, and provides the following evolutionary hypothesis for the tribes: Brachyelytrum, Nardeae, Duthieeae, Meliceae, Stipeae, Diarrheneae, Brachypodieae and the ‘core’ Pooideae. Within the individual datasets, the phylogenetic trees obtained from Topo6 exon 8–13 shows the most interesting results. The divergent positions of some clone sequences of Ampelodesmos mauritanicus and Trikeraia pappiformis, for instance, may indicate a hybrid origin of these stipoid taxa.

Karyosystematics of the Australasian stipoid grass Austrostipa and related genera: chromosome sizes, ploidy, chromosome base numbers and phylogeny

Mitotic metaphase chromosomes were counted in 29 taxa, representing 11 subgenera of Austrostipa, and in 11 species from nine related genera of the grass subfamily Pooideae. Karyotype features were also measured. The cytogenetic data were mapped on molecular phylogenetic trees based on nuclear ITS and plastid 3ʹtrnK DNA sequence data. The trees showed four different main lineages within Austrostipa, but supported only two of the 13 acknowledged subgenera. The phylogenetic positions of the genera Anemanthele, Achnatherum, Nassella and Oloptum indicated paraphyly of the genus Austrostipa. In nuclear-sequence data, Anemanthele was nested within Austrostipa; however, in plastid-sequence data, both were sisters. The newly obtained chromosome counts in Austrostipa showed that most species have 2n = 44, the other 2n = 66. Presuming a chromosome base number of x = 11, the counts corresponded with ploidy levels of 4x and 6x respectively. Karyotype data of Austrostipa and Anemanthele were very similar. Chromosome counting in further genera suggested chromosome base numbers of x = 9, 10, 11, 12 and 13. Chromosome sizes of the phylogenetically derived tribe Stipeae were smaller than those of the earliest diverging Pooideae lineages Nardeae, Meliceae and Phaenospermateae. The mechanisms of chromosome evolution and the origin of the considerable variation in chromosome base numbers in the subfamily Pooideae are discussed in the context of chromosome evolution and biosystematics.

Anther ontogeny in Brachypodium distachyon.

Brachypodium distachyon has emerged as a model plant for the improvement of grain crops such as wheat, barley and oats and for understanding basic biological processes to facilitate the development of grasses as superior energy crops. Brachypodium is also the first species of the grass subfamily Pooideae with a sequenced genome. For obtaining a better understanding of the mechanisms controlling male gametophyte development in B. distachyon, here we report the cellular changes during the stages of anther development, with special reference to the development of the anther wall. Brachypodium anthers are tetrasporangiate and follow the typical monocotyledonous-type anther wall formation pattern. Anther differentiation starts with the appearance of archesporial cells, which divide to generate primary parietal and primary sporogenous cells. The primary parietal cells form two secondary parietal layers. Later, the outer secondary parietal layer directly develops into the endothecium and the inner secondary parietal layer forms an outer middle layer and inner tapetum by periclinal division. The anther wall comprises an epidermis, endothecium, middle layer and the secretory-type tapetum. Major documented events of anther development include the degradation of a secretory-type tapetum and middle layer during the course of development and the rapid formation of U-shaped endothecial thickenings in the mature pollen grain stage. The tapetum undergoes degeneration at the tetrad stage and disintegrates completely at the bicellular stage of pollen development. The distribution of insoluble polysaccharides in the anther layers and connective tissue through progressive developmental stages suggests their role in the development of male gametophytes. Until sporogenous cell stage, the amount of insoluble polysaccharides in the anther wall was negligible. However, abundant levels of insoluble polysaccharides were observed during microspore mother cell and tetrad stages and gradually declined during the free microspore and vacuolated microspore stages to undetectable level at the mature stage. Thus, the cellular features in the development of anthers in B. distachyon share similarities with anther and pollen development of other members of Poaceae.

Arabidopsis galactinol synthase AtGolS2 improves drought tolerance in the monocot model Brachypodium distachyon

Brachypodium distachyon (purple false brome) is a herbaceous species belonging to the grass subfamily Pooideae, which also includes major crops like wheat, barley, oat and rye. The species has been established as experimental model organism for understanding and improving cereal crops and temperate grasses. The complete genome of Bd21, the community standard line of B. distachyon, has been sequenced and protocols for Agrobacterium-mediated transformation have been published. Further improvements to the experimental platform including better evaluation systems for transgenic plants are still needed. Here we describe the growth conditions for Bd21 plants yielding highly responsive immature embryos that can generate embryogenic calli for transformation. A prolonged 20-h photoperiod produced seeds with superior immature embryos. In addition, osmotic treatment of embryogenic calli enhanced the efficiency of transfection by particle bombardment. We generated transgenic plants expressing Arabidopsis thaliana galactinol synthase 2 (AtGolS2) in these experiments. AtGolS2-expressing transgenics displayed significantly improved drought tolerance, increasing with increased expression of AtGolS2. These results demonstrate that AtGolS2 can confer drought tolerance to monocots and confirm that Brachypodium is a useful model to further explore ways to understand and improve major monocot crop species.

Comparative study of transgenic Brachypodium distachyon expressing sucrose:fructan 6-fructosyltransferases from wheat and timothy grass with different enzymatic properties

Fructans can act as cryoprotectants and contribute to freezing tolerance in plant species, such as in members of the grass subfamily Pooideae that includes Triticeae species and forage grasses. To elucidate the relationship of freezing tolerance, carbohydrate composition and degree of polymerization (DP) of fructans, we generated transgenic plants in the model grass species Brachypodium distachyon that expressed cDNAs for sucrose:fructan 6-fructosyltransferases (6-SFTs) with different enzymatic properties: one cDNA encoded PpFT1 from timothy grass (Phleum pratense), an enzyme that produces high-DP levans; a second cDNA encoded wft1 from wheat (Triticum aestivum), an enzyme that produces low-DP levans. Transgenic lines expressing PpFT1 and wft1 showed retarded growth; this effect was particularly notable in the PpFT1 transgenic lines. When grown at 22°C, both types of transgenic line showed little or no accumulation of fructans. However, after a cold treatment, wft1 transgenic plants accumulated fructans with DP=3-40, whereas PpFT1 transgenic plants accumulated fructans with higher DPs (20 to the separation limit). The different compositions of the accumulated fructans in the two types of transgenic line were correlated with the differences in the enzymatic properties of the overexpressed 6-SFTs. Transgenic lines expressing PpFT1 accumulated greater amounts of mono- and disaccharides than wild type and wft1 expressing lines. Examination of leaf blades showed that after cold acclimation, PpFT1 overexpression increased tolerance to freezing; by contrast, the freezing tolerance of the wft1 expressing lines was the same as that of wild type plants. These results provide new insights into the relationship of the composition of water-soluble carbohydrates and the DP of fructans to freezing tolerance in plants.

Evidence for adaptive evolution of low‐temperature stress response genes in a Pooideae grass ancestor

Adaptation to temperate environments is common in the grass subfamily Pooideae, suggesting an ancestral origin of cold climate adaptation. Here, we investigated substitution rates of genes involved in low-temperature-induced (LTI) stress responses to test the hypothesis that adaptive molecular evolution of LTI pathway genes was important for Pooideae evolution.Substitution rates and signatures of positive selection were analyzed using 4330 gene trees including three warm climate-adapted species (maize (Zea mays), sorghum (Sorghum bicolor), and rice (Oryza sativa)) and five temperate Pooideae species (Brachypodium distachyon, wheat (Triticum aestivum), barley (Hordeum vulgare), Lolium perenne and Festuca pratensis).Nonsynonymous substitution rate differences between Pooideae and warm habitat-adapted species were elevated in LTI trees compared with all trees. Furthermore, signatures of positive selection were significantly stronger in LTI trees after the rice and Pooideae split but before the Brachypodium divergence (P < 0.05). Genome-wide heterogeneity in substitution rates was also observed, reflecting divergent genome evolution processes within these grasses.Our results provide evidence for a link between adaptation to cold habitats and adaptive evolution of LTI stress responses in early Pooideae evolution and shed light on a poorly understood chapter in the evolutionary history of some of the world's most important temperate crops.

Early inflorescence development in the grasses (Poaceae)

The shoot apical meristem of grasses produces the primary branches of the inflorescence, controlling inflorescence architecture and hence seed production. Whereas leaves are produced in a distichous pattern, with the primordia separated from each other by an angle of 180°, inflorescence branches are produced in a spiral in most species. The morphology and developmental genetics of the shift in phyllotaxis have been studied extensively in maize and rice. However, in wheat, Brachypodium, and oats, all in the grass subfamily Pooideae, the change in phyllotaxis does not occur; primary inflorescence branches are produced distichously. It is unknown whether the distichous inflorescence originated at the base of Pooideae, or whether it appeared several times independently. In this study, we show that Brachyelytrum, the genus sister to all other Pooideae has spiral phyllotaxis in the inflorescence, but that in the remaining 3000+ species of Pooideae, the phyllotaxis is two-ranked. These two-ranked inflorescences are not perfectly symmetrical, and have a clear “front” and “back;” this developmental axis has never been described in the literature and it is unclear what establishes its polarity. Strictly distichous inflorescences appear somewhat later in the evolution of the subfamily. Two-ranked inflorescences also appear in a few grass outgroups and sporadically elsewhere in the family, but unlike in Pooideae do not generally correlate with a major radiation of species. After production of branches, the inflorescence meristem may be converted to a spikelet meristem or may simply abort; this developmental decision appears to be independent of the branching pattern.

Stipa (Poaceae) and allies in the Old World: molecular phylogenetics realigns genus circumscription and gives evidence on the origin of American and Australian lineages

The tribe Stipeae with an estimated number of ca. 600 species is part of the grass subfamily Pooideae and has near worldwide distribution. Its species are often dominant constituents of steppe vegetation and other grasslands, especially in Eurasia, the Americas and Australia. The taxonomy of Old World Stipeae has been studied to date primarily on the basis of morphology and anatomy, while existing molecular phylogenetic investigations have mainly dealt with New World or Australian taxa. We studied 109 new ingroup taxa with a focus on Old World Stipeae (in addition with an extensive outgroup sampling) using chloroplast and nuclear ribosomal DNA sequences (3′trnK region, ITS1–5.8S gene–ITS2), and discuss taxonomic key characters. Five highly supported monophyletic lineages were identified, some of which were rather unexpected: (a) the narrowly defined Stipa core clade, which is primarily Eurasian but extends into Africa north of the Sahara Desert, (b) the majority of Old World Piptatherum, (c) a “Transcontinental Stipeae Clade” encompassing Eurasian, African, American and Australian lineages, (d) a Himalayan to E Asian clade and (e) the single species Achnatherum splendens. The large “Transcontinental Stipeae Clade” contained several lineages of Eurasian Stipeae different from the Stipa core (a), i.e., genera Aristella, Celtica, Oloptum gen. nov., Stipella stat. et. gen. nov., species of Achnatherum, and the species-rich lineages of Nassella/Jarava in America and of Austrostipa in Australia. In our circumscription Ptilagrostis was nested in (d), a clade (which included some species of Achnatherum and poorly studied Himalayan species ascribed to either Stipa or Orthoraphium) and whose internal structure remained unclear. Oloptum, gen. nov., is described, and the following combinations are made: Achnatherum pelliotii, comb. nov., Aristella keniensis, comb. nov., Oloptum miliaceum, comb. nov., Stipella, stat. et. gen. nov., S. capensis, comb. nov., S. nitens, comb. nov., S. parviflora, comb. nov., S. staintonii, comb. nov., and S. tigrensis, comb. nov.

Did gene family expansions during the Eocene-Oligocene boundary climate cooling play a role in Pooideae adaptation to cool climates?

Adaptation to cool environments is a common feature in the core group of the grass subfamily Pooideae (Triticeae and Poeae). This suggest an ancient evolutionary origin of low temperature stress tolerance dating back prior to the initiation of taxonomic divergence of core Pooideae species. Viewing the Pooideae evolution in a palaeo-climatic perspective reveals that taxonomic divergence of the core Pooideae group initiated shortly after a global super-cooling period at the Eocene-Oligocene boundary (approximately 33.5-26 Ma). This global climate cooling altered distributions of plants and animals and must have imposed selection pressure for improved low temperature stress responses. Lineage-specific gene family expansions are known to be involved in adaptation to new environmental stresses. In Pooideae, two gene families involved in low temperature stress response, the C-repeat binding factor (CBF) and fructosyl transferase (FT) gene families, has undergone lineage-specific expansions. We investigated the timing of these gene family expansions by molecular dating and found that Pooideae-specific expansion events in CBF and FT gene families took place during Eocene-Oligocene super-cooling period. We hypothesize that the E-O super-cooling exerted selection pressure for improved low temperature stress response and frost tolerance in a core Pooideae ancestor, and that those individuals with multiple copies of CBF and FT genes were favoured.

Phylogenetic structure of the grass subfamily Pooideae based on comparison of plastid matK gene–3′trnK exon and nuclear ITS sequences

ABSTRACTThe grass subfamily Pooideae was studied using DNA sequence information from the chloroplast (cp) matK gene–3′trnK exon and the nuclear ribosomal (nr) ITS1–5.8S gene–ITS2 in a sample of 67 taxa covering all of its tribes. Branches with strong bootstrap support are consistently resolved in both datasets, whereas discrepancy is confined to low‐support or unsupported nodes in one of the datasets. The results do not reveal a significant role of past hybridisation, plastid lineage sorting or reticulation in the evolutionary diversification of the major lineages of the subfamily. The combined analysis of the plastid and nuclear datasets results in a largely well‐supported pattern of divergence for the major lineages of the subfamily. Some re‐alignments of tribes and subtribes are proposed and discussed with reference to relevant morphological and structural characters. We propose the recognition of broader tribes Nardeae with subtribes Nardinae and Lygeinae, Meliceae with subtribes Brylkiniinae and Melicinae, Stipeae with subtribes Ampelodesminae and Stipinae, and Triticeae with subtribes Littledaleinae, Brominae and Hordeinae. For the tribe complex of Aveneae and Poeae, the clear‐cut split into two major clades and further resolution into some high‐support lineages depicted by cpDNA is not contradicted by nuclear ITS and their taxonomic treatment as separate tribes or a single tribe remains an unanswered question.

Ice recrystallization inhibition proteins (IRIPs) and freeze tolerance in the cryophilic Antarctic hair grass Deschampsia antarctica E. Desv.

ABSTRACTAntarctic hair grass (Deschampsia antarctica E. Desv.), the only grass indigenous to Antarctica, has well‐developed freezing tolerance, strongly induced by cold acclimation. Here, we show that in response to low temperatures, D. antarctica expresses potent recrystallization inhibition (RI) activity that, inhibits the growth of small ice crystals into potentially damaging large ones, is proteinaceous and localized to the apoplasm. A gene family from D. antarctica encoding putative homologs of an ice recrystallization inhibition protein (IRIP) has been isolated and characterized. IRIPs are apoplastically targeted proteins with two potential ice‐binding motifs: 1–9 leucine‐rich repeats (LRRs) and c. 16 ‘IRIP’ repeats. IRIP genes appear to be confined to the grass subfamily Pooideae and their products, exhibit sequence similarity to phytosulphokine receptors and are predicted to adopt conformations with two ice‐binding surfaces. D. antarctica IRIP (DaIRIP) transcript levels are greatly enhanced in leaf tissue following cold acclimation. Transgenic Arabidopsis thaliana expressing a DaIRIP has novel RI activity, and purified DaIRIP, when added back to extracts of leaves from non‐acclimated D. antarctica, can reconstitute the activity found in acclimated plants. We propose that IRIP‐mediated RI activity may contribute to the cryotolerance of D. antarctica, and thus to its unique ability to have colonized Antarctica.