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Repeated DNA sequences of Aegilops markgrafii (Greuter) Hammer var. markgrafii: cloning, sequencing and analysis of distribution in Poaceae species.

Total DNA of Aegilops markgrafii (Greuter) Hammer var. markgrafii was shot gun cloned. From all the recombinants containing repetitive sequences 1-2% hybridized preferentially with the Ae. markgrafii genome and were almost absent in wheat. The cloned sequences are disperse distributed over the Aegilops chromosomes and show the typical features of eukaryotic repetitive DNA. Five specific probes were tested for their applicability in a screening program on 68 Poaceae accessions.

Base Sequence↗

16-kilodalton rice protein is one of the major allergens in rice grain extract and responsible for cross-allergenicity between cereal grains in the Poaceae family.

Cross-allergenicity between five cereal grains including rice, wheat, corn, Japanese millet (Panicum crus-galli L. var. frumentaceum Trin.) and Italian millet (Setaria italica Beauv. var. germanica schrad.) was examined by radioallergosorbent test (RAST) and RAST inhibition assay. There were significant close correlations between every combinations of RAST values for the five cereal grain extracts. RAST inhibition assay of each extract against RAST discs coupled with other cereal grain extracts indicated marked cross-reactivity of IgE binding between these cereal grain extracts. Rice protein 16KD (RP16KD) was shown to be one of major allergens in rice grain extracts by immunoblotting analysis, histamine release assay from human leukocytes and RAST inhibition. Next, the involvement of RP16KD in the cross-allergenicity between these cereals was investigated. RAST values for RP16KD significantly correlated with that for Italian millet as well as rice but not with those for corn and wheat. There was a trend of positive correlation between RAST values for RP16KD and Japanese millet. In the RAST inhibition assay using sera with positive RAST for these five cereal grain extracts and RP16KD, RP16KD inhibited IgE binding to these all cereal discs in a dose-dependent manner. Similarly, all of the five cereal grain extracts showed an effective decrease in IgE binding to the RP16KD disc. These results indicated possible participation of IgE binding structure on RP16KD in cross-allergenicity between these cereal grain extracts in the Poaceae family.

Allergens↗

Genome structure and evolution in the allohexaploid weed Avena fatua L. (Poaceae).

Allohexaploid wild oat, Avena fatua L. (Poaceae; 2n = 6x = 42), is one of the world's worst weeds, yet unlike some of the other Avena hexaploids, its genomic structure has been relatively little researched. Consequently, in situ hybridisation was carried out on one accession of A. fatua using an 18S-25S ribosomal DNA (rDNA) sequence and genomic DNA from A. strigosa (AA-genome diploid) and A. clauda (CC-genome diploid) as probes. Comparing these results with those for other hexaploids studied previously: (i) confirmed that the genomic composition of A. fatua was similar to the other hexaploid Avena taxa (i.e., AACCDD), (ii) identified major sites of rDNA on three pairs of A/D-genome chromosomes, in common with other Avena hexaploids, and (iii) revealed eight chromosome pairs carrying intergenomic translocations between the A/D- and C-genomes in the accession studied. Based on karyotype structure, the identity of some of these recombinant chromosomes was proposed, and this showed that some of these could be divided into two types, (i) those common to all hexaploid Avena species analysed (3 translocations) and (ii) one translocation in this A. fatua accession not previously observed in reports on other hexaploid Avena species. If this translocation is found to be unique to A. fatua, then this information, combined with more traditional morphological data, will add support to the view that A. fatua is genetically distinct from other hexaploid Avena species and thus should retain its full specific status.

Avena↗

Phylogenetic structure in the grass family (Poaceae): evidence from the nuclear gene phytochrome B.

Phylogenetic analyses of partial phytochrome B (PHYB) nuclear DNA sequences provide unambiguous resolution of evolutionary relationships within Poaceae. Analysis of PHYB nucleotides from 51 taxa representing seven traditionally recognized subfamilies clearly distinguishes three early-diverging herbaceous "bambusoid" lineages. First and most basal are Anomochloa and Streptochaeta, second is Pharus, and third is Puelia. The remaining grasses occur in two principal, highly supported clades. The first comprises bambusoid, oryzoid, and pooid genera (the BOP clade); the second comprises panicoid, arundinoid, chloridoid, and centothecoid genera (the PACC clade). The PHYB phylogeny is the first nuclear gene tree to address comprehensively phylogenetic relationships among grasses. It corroborates several inferences made from chloroplast gene trees, including the PACC clade, and the basal position of the herbaceous bamboos Anomochloa, Streptochaeta, and Pharus. However, the clear resolution of the sister group relationship among bambusoids, oryzoids, and pooids in the PHYB tree is novel; the relationship is only weakly supported in ndhF trees and is nonexistent in rbcL and plastid restriction site trees. Nuclear PHYB data support Anomochlooideae, Pharoideae, Pooideae sensu lato, Oryzoideae, Panicoideae, and Chloridoideae, and concur in the polyphyly of both Arundinoideae and Bambusoideae.

Journal Article↗

Analysis of inflorescence organogenesis in eastern gamagrass, Tripsacum dactyloides (Poaceae): the wild type and the gynomonoecious gsf1 mutant.

Inflorescence organogenesis of a wild-type and a gynomonoecious (pistillate) mutant in Tripsacum dactyloides was studied using scanning electron microscopy. SEM (scanning electron microscope) analysis indicated that wild-type T. dactyloides (Eastern gamagrass) expressed a pattern of inflorescence organogenesis that is observed in other members of the subtribe Tripsacinae (Zea: maize and teosinte), family Poaceae. Branch primordia are initiated acropetally along the rachis of wild-type inflorescences in a distichous arrangement. Branch primordia at the base of some inflorescences develop into long branches, which themselves produce an acropetal series of distichous spikelet pair primordia. All other branch primordia function as spikelet pair primordia and bifurcate into pedicellate and sessile spikelet primordia. In all wild-type inflorescences development of the pedicellate spikelets is arrested in the proximal portion of the rachis, and these spikelets abort, leaving two rows of solitary sessile spikelets. Organogenesis of spikelets and florets in wild-type inflorescences is similar to that previously described in maize and the teosintes. Our analysis of gsf1 mutant inflorescences reveals a pattern of development similar to that of the wild type, but differs from the wild type in retaining (1) the pistillate condition in paired spikelets along the distal portion of the rachis and (2) the lower floret in sessile spikelets in the proximal region of the rachis. The gsf1 mutation blocks gynoecial tissue abortion in both the paired-spikelet and the unpaired-spikelet zone. This study supports the hypothesis that both femaleness and maleness in Zea and Tripsacum inflorescences are derived from a common developmental pathway. The pattern of inflorescence development is not inconsistent with the view that the maize ear was derived from a Tripsacum genomic background.

Journal Article↗

Genomic and genetic relationships among species of Leymus (Poaceae: Triticeae) inferred from 18S-26S ribosomal genes.

The 18S-26S ribosomal genes in three closely related species of Leymus (Poaceae: Triticeae) were examined using fluorescence in situ hybridization (FISH) and restriction fragment length polymorphism (RFLP). Both approaches revealed a close relationship between L. arenarius (8x = 56, northern European) and L. racemosus (4x = 28, central Eurasian), whereas L. mollis (4x = 28, northern American/Pacific) was distinct. Each species had three homologous pairs of major rDNA loci: a1, a2, and a3 for L. arenarius; m1, m2, and m3 for L. mollis; and r1, r2, and r3 for L. racemosus. Leymus arenarius had in addition three minor loci, a4, a5, and a6. The major loci of L. arenarius and L. racemosus were identical, indicating that the former species could have originated from the latter, via interspecific hybridization and/or polyploidy. The rDNA-RFLPs further indicated relationships of these species to other species of Leymus (L. karellini, 8x = 56 and L. angustus, 12x = 84) and Psathyrostachys (P. fragilis, P. huashanica, P. juncea, and P. lanuginosa, which are all diploids). A phenogram constructed from 20 BamHI, EcoRI, and DraI rDNA fragments revealed closer relationship between the two genera, Leymus and Psathyrostachys, than that among species within a genus.

Journal Article↗

An easy and rapid method using microscopy to determine herbicide effects in Poaceae weed species.

A new, easy, rapid and relatively inexpensive method using microscopy has been developed for the detection of herbicide effects in leaves of grass weed species displaying no visual signs of damage. The method has potential to be used as a tool to indicate future death of grass species due to herbicide exposure by observing phytochemical effects, i.e. early-warning effects. In the present study, Apera spica-venti (L.) Beauv., Bromus hordeaceus L., Alopecurus myosuroides Huds., Lolium perenne L. and Poa annua L. were exposed to lethal rates of four herbicides with different mode of action. The herbicides investigated were the thiocarbamate: prosulfocarb, the sulfonylurea: iodosulfuron, the aryloxyphenoxypropionate: fenoxaprop-P-ethyl and the organophosphate glyphosate. Autofluorescence of leaves was studied under a microscope using ultraviolet and blue light. The fluorescence of leaves treated to enhance flavonoids was also examined. To confirm the results, microspectrofluorometry was performed. Effects indicating future death of the grasses were observed in visually healthy leaves following treatment with prosulfocarb, glyphosate and iodosulfuron. No changes were detected following treatment with fenoxaprop-P-ethyl. After exposure to glyphosate or iodosulfuron, changes in the content of flavonoids and other compounds with a conjugation system and rigid structure and a decrease in the content of chlorophyll were detected in the leaves. Prosulfocarb treatment resulted in changes in the content of flavonoids and other compounds with a conjugation system and rigid structure and an increase in the content of chlorophyll in the leaves. The results obtained from microspectrofluorometry indicated that exposure to prosulfocarb caused a reduction in the flavonoids quercetin, naringenin and/or naringin.

Drug Resistance↗

Phylogenetic analysis of the Triticeae (Poaceae) based on rpoA sequence data.

A phylogenetic analysis was conducted on 31 diploid species representing 21 of the 24 monogenomic genera of the Triticeae. The data used were derived from a 1343- to 1358-bp region of the plastid genome spanning the entire rpoA gene plus minor parts of the petD and rps11 genes and the two intergenic spacers surrounding rpoA. Bromus inermis (Bromeae) was used as an outgroup. A total of 68 variable sites, 25 of them phylogenetically informative, and seven length mutations were detected. The length mutations occurred in the noncoding regions. Phylogenetic analyses were performed on the whole data set and on various subsets. The analysis of the unweighted data resulted in 48 equally parsimonious trees (length 98, CI = 0.88, RI = 0.92, ti = 0.25). A parsimony jackknife analysis proved several clades to be well supported. The effect of transition/transversion weighting was also investigated. In general, congruence with other data sets was negatively effected by weighting. The preferred phylogenetic hypothesis is congruent with a phylogeny based on plastid RFLP data including the same taxa, but it is largely incongruent with phylogenies derived from nuclear rDNA and morphology.

Base Sequence↗

Phylogeny of the grass family (Poaceae) from rpl16 intron sequence data.

DNA sequence data from the chloroplast noncoding rpl16 intron are used to address phylogenetic relationships among the major lineages of the grass family, with particular emphasis on the highly heterogeneous subfamily Bambusoideae and the basal lineages. Thirty-five grass sequences representing all six currently recognized major groups of the family and one outgroup sequence were analyzed using both parsimony and distance methods. The phylogenetic analyses indicated: (1) Puelia, a traditionally isolated bambusoid genus, is the most basal lineage in the BOP clade (Bambusoideae, Oryzoideae, and Pooideae); (2) the bambusoid clade is a sister group to the pooid clade; and (3) the monophyletic oryzoid clade is well separated from the bambusoid clade. The study further confirmed the recognition of two primary groups in the grass family: the BOP clade and the PACC clade (Panicoideae, Arundinoideae, Chloridoideae, and Centothecoideae); it also provided further evidence that the traditional subfamily Bambusoideae is highly heterogeneous and phylogenetically unacceptable. The data support Streptochaeteae, Anomochloeae, and Phareae as the most basal lineages among the extant grasses. Within the BOP clade, oryzoids and pooids are confirmed as two monophyletic clades, but the bambusoid clade, including only the woody bamboo tribe Bambuseae and the herbaceous bamboo tribe Olyreae, is relatively weakly supported. The study also indicated that the chloroplast noncoding region sequence data could be useful in phylogenetic analysis at relatively high taxonomic levels.

Chloroplasts↗

Editing of the chloroplast ndhB encoded transcript shows divergence between closely related members of the grass family (Poaceae).

The ndhB-encoded transcript from barley chloroplasts deviates from the genomic ndhB sequence by nine C-to-U transitions, which is the maximum number of editing events for a chloroplast mRNA reported so far. Comparison with ndhB transcripts from other chloroplast species shows that six of the nine editing sites observed in barley are structurally and functionally conserved in maize, rice and tobacco. The remaining three sites, however, show divergent patterns of conservation even within the three members of the grass family. The conservation of two of these sites in tobacco but not in the closely related graminean species suggests that divergence of the ndhB editing sites is caused by the loss of preexisting editing sites rather than by gain of new sites.

Amino Acid Sequence↗

The chromosomes of Festuca pratensis Huds. (Poaceae): fluorochrome banding, heterochromatin and condensation.

The karyotype of meadow fescue Festuca pratensis Huds. (2n = 14) has been studied by means of fluorescence banding techniques. AT-enriched heterochromatic bands were revealed in the metaphase chromosomes 1-6. A single GC-rich band was associated with the chromosome 2 secondary constriction. Delay of condensation of the heterochromatic regions was discovered. The heterochromatic bands reported are markers which allow identification of all meadow fescue chromosomes.

Chromosome Banding↗

Characterisation of extracellular polysaccharides from suspension cultures of members of the poaceae.

Microscopic examination of suspension- cultured cells of Phleum pratense L., Panicum miliaceum L., Phalarisaquatica L. and Oryza sativa L. showed that they were comprised of numerous root primordia. Polysaccharides secreted by these suspension cultures contained glycosyl linkages consistent with the presence of high proportions of root mucilage-like polysaccharides. In contrast, suspension-cultured cells of Hordeum vulgare L. contained mostly undifferentiated cells more typical of plant cells in suspension culture. The polysaccharides secreted by H. vulgare cultures contained mostly linkages consistent with the presence of glucuronoarabinoxylan. The soluble polymers secreted by cell-suspension cultures of Phleum pratense contained 70% carbohydrate, 14% protein and 6% inorganic material. The extracellular polysaccharides were separated into four fractions by anion-exchange chromatography using a gradient of imidazole-HCl at pH 7.0. From glycosyl-linkage analyses, five polysaccharides were identified: an arabinosylated xyloglucan (comprising 20% of the total polysaccharide), a glucomannan (6%), a type-II arabinogalactan (an arabinogalactan-protein; 7%), an acidic xylan (3%), and a root-slime-like polysaccharide, which contained features of type-II arabinogalactans and glucuronomannans (65%).

Cell Wall↗

Structural and distributional variation of mitochondrial rps2 genes in the tribe Triticeae (Poaceae).

The mitochondrial rps2 gene from barley, like that of rice, wheat, and maize, has an extended open reading frame (ORF) at the 3'-region when compared to that from lower plants. However, the extended portions are variable among these cereals. Since barley and wheat belong to the same tribe (Triticeae), it would be interesting to know when and where the two types of rps2 were generated during evolution. To determine this, we utilized the mitochondrial (mt) DNA sequence to examine variations of the rps2 genes in the tribe Triticeae. By means of the variable 3'-region, the distribution of barley (B)-type and wheat (W)-type rps2 sequences was studied in 19 genera of the tribe. The B-type sequence was identified in 10 of the 19 genera, whereas the W-type sequence was present in all 19 genera. Thus, ten of the examined genera have both types of rps2 sequences due to the presence of two copies of the gene. The W-type sequence was also present in the tribe Bromeae and the B-type sequence was also found in Aveneae and Poeae. Phylogenetic trees based on the B-type and W-type sequences were different from those based on other molecular data. This suggests that the mitochondrial genome in Triticeae has a unique evolutionary history.

Amino Acid Sequence↗

An AFLP-based survey of genetic diversity among accessions of sea oats (Uniola paniculata, Poaceae) from the southeastern Atlantic and Gulf coast states of the United States.

Uniola paniculata, commonly known as sea oats, is a C4 perennial grass capable of stabilizing sand dunes. It is most abundant along the Gulf of Mexico and southeastern Atlantic coastal regions of the United States. The species exhibits low seed set and low rates of germination and seedling emergence, and so extensive clonal reproduction is achieved through production of rhizomes, which may contribute to a decline in genetic diversity. To date, there has been no systematic assessment of genetic variability and population structure in naturally occurring stands in the USA. This study was conducted to assess the genetic relationship and diversity among nineteen U. paniculata accessions representing eight states: Texas, Louisiana, Mississippi, Alabama, Florida, South Carolina, North Carolina, and Virginia, using amplified fragment length polymorphism (AFLP). Twelve AFLP EcoRI + MseI primer combinations generated a wide range of polymorphisms (42-81%) with a mean of 59%. Overall, the sea oats plants exhibited a low range of genetic similarity. Florida accessions, FL-33 and FL-39, were most genetically diverse and the accessions from both Carolinas and Virginia (NC-1, NC-11, SC-15, and VA-53) harbored less genetic variability. Cluster analysis using the UPGMA approach separated U. paniculata plants into four major clusters which were also confirmed by principal coordinate analysis (PCO). Further examination of the different components of genetic variation by analysis of molecular variance (AMOVA) indicated the largest proportion of variability at the state level (47.8%) followed by the variation due to the differences among the genotypes within an accession (34.4%), and the differences among the accessions within a state (17.8%). The relationship between genetic diversity and geographic source of sea oats populations of the United States as revealed through this comprehensive study will be helpful to resource managers and commercial nurseries in identifying suitable plant materials for restoration of new areas without compromising the adaptation and genetic diversity.

Analysis of Variance↗

Taxonomic significance of leaf anatomy of Aniselytron (Poaceae) as an evidence to support its generic validity against Calamagrostis s. l.

A comparative study of leaf anatomy on Aniselytron Merr. and Calamagrostis Adans. s. l. was conducted to review the systematic status of Aniselytron Merr. Calamagrostis s. l. exhibits wide variation in many features, but basic leaf structures of the genus remain constant: absence of a midrib-keel; median and large vascular bundles are central, with double sheaths, accompanied by girders both adaxially and abaxially; prickles have a bulbous base and are not sunken; the abaxial epidermal cells are short and wide and relatively thick-walled. Aniselytron differs from Calamagrostis s. l. in: midrib-keel is present, consisting of a large central bundle with small ones on either side; all vascular bundles are abaxially situated, with abaxial girders only, parenchyma takes the place of the adaxial sclerenchyma; the bases of the prickles are sunken and are not bulbous; the abaxial epidermal cells are tall and thin-walled. These distinct anatomical features, in combination with the differences in spikelet structure and habitat, suggest that Aniselytron should be generically separated from and not merged with Calamagrostis s. l. Due to the adaxial parenchyma in the midrib which has never been found in Pooideae, Aniselytron might have a relationship with some other subfamily.

Plant Epidermis↗

Genome constitutions of Hystrix patula, H. duthiei ssp. duthiei and H. duthiei ssp. longearistata (Poaceae: Triticeae) revealed by meiotic pairing behavior and genomic in-situ hybridization.

Genomic constitutions of three taxa of Hystrix Moench, H. patula, H. duthiei ssp. duthiei and H. duthiei ssp. longearistata, were examined by meiotic pairing behavior and genomic in-situ hybridization. Meiotic pairing in hybrids of H. patula x Pseudoroegneria spicata (St), H. patula x Elymus wawawaiensis (StH), H. patula x H. duthiei ssp. longearistata, H. patula x Psathyrostachys huashanica (Ns ( h )), H. duthiei ssp. duthiei x Psa. huashanica, H. duthiei ssp. longearistata x Psa. huashanica, Leymus multicaulis (NsXm) x H. duthiei ssp. longearistata averaged 6.53, 12.83, 1.32, 0.29, 5.18, 5.11 and 10.47 bivalents per cell, respectively. The results indicate that H. patula has the StH genome and H. duthiei ssp. duthiei and H. duthiei ssp. longearistata have the NsXm genome. Results of genomic in-situ hybridization analysis strongly supported the chromosome pairing data; therefore it is concluded that the type species of Hystrix, H. patula, should be included in Elymus, and that H. duthiei ssp. duthiei and H. duthiei ssp. longearistata should be transferred to Leymus.

Chromosome Pairing↗

Molecular evolution of the maize sex-determining gene TASSELSEED2 in Bouteloua (Poaceae).

The majority of angiosperms produce hermaphrodite flowers, while a lesser number (20-30%) produce unisexual flowers. Little is known about the molecular biology of sex-determination in angiosperms, however, a few sex-determining genes have been cloned from the model system Zea mays. One of these genes is Tasselseed2 (Ts2) which has been shown to be involved in the arrest of developing pistils in male flowers. In this study, we sequenced a putative homologue of Ts2 in species of Bouteloua, a genus in the grass subfamily Chloridoideae. We found significant genetic variation at Ts2 in Bouteloua relative to other developmental genes characterized in maize and other grass species. We also found that in Bouteluoua, Ts2 is evolving non-neutrally in the hermaphrodite-flowered Bouteloua hirsuta while no difference from neutral expectation was detected at Ts2 in the monoecious/dioecious Bouteloua dimorpha. The putatively neutral gene Alcohol Dehydrogenase1 (Adh1) was also examined for the same species of Bouteloua, and no departure from neutral expectation was detected. Our results suggest that purifying selection may be acting on Ts2 in the hermaphrodite-flowered B. hirsuta while no evidence of selection was detected at Ts2 in the monoecious/dioecious B. dimorpha.

Base Sequence↗

Molecular and morphological phylogenetic analysis of Brachiaria and Urochloa (Poaceae).

The taxonomic relationships of Brachiaria and Urochloa have been questioned based on previous morphological studies. In this paper, we reconsider the phylogenetic relationships of these genera using 22 species of Brachiaria and Urochloa and six species of Paniceae as out-groups. The ITS1, 5.8S, and ITS2 region (internal transcribed spacer) of nuclear ribosomal DNA and eight morphological characters of the inflorescence were compiled into a data matrix. The cladistic analyses suggest that Urochloa-Brachiaria as a complex is paraphyletic with Eriochloa and Melinis. Species of all these genera share molecular synapomorphies and belong to the same monophyletic groups. The results confirm the continuous gradation between those genera previously found in several morphological studies. Therefore, the following eight new combinations are made: Urochloa bovonei (Chiov.) A.M. Torres & C.M. Morton, Urochloa dura (Stapf) A.M. Torres & C.M. Morton, Urochloa dura var. dura (Stapf) A.M. Torres & C.M. Morton, Urochloa dura var. pilosa (J.G. Anderson) A.M. Torres & C.M. Morton, Urochloa lachnantha (Hochst.) A.M. Torres & C.M. Morton, Urochloa leersioides (Hochst.) A.M. Torres & C.M. Morton, Urochloa nigropedata (Munro ex Ficalho & Hiern) A.M. Torres & C.M. Morton, and Urochloa subulifolia (Mez) A.M. Torres & C.M. Morton.

Base Sequence↗