Genetic Differentiation of Reed Canarygrass (Phalaris arundinacea L.) within Eastern Europe and Eurasia.

Worldwide molecular research of economically important Phalaris arundinacea (Poaceae) is mainly focused on the invasions of this species from Europe to North America. Until the present study, the genetic diversity of the P. arundinacea had not been studied across the Baltic countries. The objective of this research is to evaluate the diversity of Lithuanian populations of P. arundinacea at simple sequence repeat (SSR) loci comparatively among populations of the Baltic countries, Luxembourg, and the Russian Far East (Eurasian), evaluating differentiation between Lithuanian populations and ornamental accessions, and relating these with environmental features. For six selected Lithuanian river basin populations, GBS low density SNPs were used to determine genetic diversity. Bayesian analysis showed that Eurasian populations of Phalaris arundinacea consist of two gene clusters. Statistically significant genetic differentiation among European and Eurasian populations was documented. Lithuanian genotypes growing naturally along rivers are genetically distinct from cultivated ornamentals. GBS-SNPs divided the six selected Nemunas river basins into three distinct groups with one, two, or three rivers in separate groupings for genetic diversity. Genetic diversity is primarily within, rather than among, Lithuanian, eastern European, and Eurasian populations of P. arundinacea across the continent. Thus, restoration efforts would benefit from local population seed origination.

Physiological and molecular responses of Phalaris arundinacea under salt stress on the Tibet plateau.

Phalaris arundinacea, with its characteristics of rapid growth and high biological yield, is regarded as an excellent forage grass in the Qinghai-Tibetan Plateau region of China. To explore the physiological and molecular response mechanism of Phalaris arundinacea under salt stress, we monitored the biomass and physiological indexes of two locally grown strains under conditions of exposure to 150 and 300 mM NaCl solution. Z0611 exhibited better salt stress tolerance than YS. Transcriptome sequencing analysis showed that YS and Z0611 had 1713 and 4290 differentially expressed genes (DEGs), respectively, including on metabolic processes, single-organism process, catalytic activity, and plant hormone signal transduction in the GO and KEGG databases. We also identified a large number of genes involved in hormone signaling, antioxidant systems, ion homeostasis, and photosynthetic systems. Our study provides physiological and molecular insight for establishing a salt resistance database and mining salt tolerance genes in Phalaris arundinacea, and also provides theoretical guidance for the restoration of saline-alkali land on the Qinghai-Tibet Plateau.

Variability in ITS1 and ITS2 sequences of historic herbaria and extant (fresh) Phalaris species (Poaceae).

BACKGROUND: Phalaris species (Poaceae) occupy diverse environments throughout all continents except Antarctica. Phalaris arundinacea is an important forage, ornamental, wetland restoration and biofuel crop grown globally as well as being a wetland invasive. The nuclear ribosomal internal transcribed spacer (ITS) region has been used for Phalaris barcoding as a DNA region with high nucleotide diversity for Phalaris species identification. Recent findings that P. arundinacea populations in Minnesota USA are most likely native and not European prompted this analysis to determine whether Eurasian vs. native North American P. arundinacea differed in ITS regions. Our objectives were to amplify and compare ITS regions (ITS1 and ITS2) of historic herbaria (1882-2001) and extant (fresh) Phalaris specimens; analyze ITS regions for species-specific polymorphisms (diagnostic SNPs) and compare ITS regions of historic Phalaris specimens with known, extant Phalaris species. RESULTS: We obtained complete ITS1 and ITS2 sequences from 31 Phalaris historic (herbaria samples, 1908 to 2001) and five extant (fresh) specimens. Herbaria Phalaris specimens did not produce new SNPs (single nucleotide polymorphisms) not present in extant specimens. Diagnostic SNPs were identified in 8/12 (66.6%) Phalaris species. This study demonstrates the use of herbaria tissue for barcoding as a means for improved species identification of Phalaris herbaria specimens. No significant correlation between specimen age and genomic DNA concentration was found. Phalaris arundinacea showed high SNP variation within its clade, with the North American being distinctly different than other USA and most Eurasian types, potentially allowing for future identification of specific SNPs to geographic origin. CONCLUSIONS: While not as efficient as extant specimens to obtain DNA, Phalaris herbaria specimens can produce high quality ITS sequences to evaluate historic genetic resources and facilitate identification of new species-specific barcodes. No correlation between DNA concentration and age of historic samples (119 year range) occurred. Considerable polymorphism was exhibited in the P. arundinacea clade with several N. American accessions being distinct from Eurasian types. Further development of within species- and genus-specific barcodes could contribute to designing PCR primers for efficient and accurate identification of N. American P. arundinacea. Our finding of misidentified Phalaris species indicates the need to exercise stringent quality control measures on newly generated sequence data and to approach public sequence databases in a critical way.

Physiological and transcriptional responses of Phalaris arundinacea under waterlogging conditions.

As a high-yielding forage grass, Phalaris arundinacea widely distributed in the Qinghai-Tibet Plateau region of China. To explore physiological and molecular response mechanism of Phalaris arundinacea under waterlogging, we analyzed the biomass and physiological indexes of three locally grown strains under the submerged condition of 10 cm. The material Z0611 showed the strongest waterlogging resistance while the YS showed the weakest performance. Transcriptome sequencing analysis demonstrated that the YS and Z0611 had 17010 and 7566 differently expression genes (DEGs), respectively, which were mainly concentrated in the metabolic process, cell, ribosome, phenylpropanoid biosynthesis pathway in GO and KEGG databases. We also identified a large number of genes involved in carbohydrate metabolism, hormone signaling regulation, transcription factors, antioxidant system, and ethylene signaling. Our research may provide a scientific basis for the restoration of wetland environment on the Qinghai-Tibet Plateau, and lay a foundation for further exploration of the waterlogging resistance genes of Phalaris arundinacea and breeding of new strains resistant with waterlogging stress.

Cross-resistance mechanisms to ACCase-inhibiting herbicides in short-spike canarygrass (Phalaris brachystachys).

Herbicides that inhibit acetyl-coenzyme A carboxylase (ACCase) are commonly used to control weedy grasses such as short-spike canarygrass (Phalaris brachystachys). Two resistant biotypes of P. brachystachys (R1 and R2) were found in different winter wheat fields in Iran. This study was done to confirm the suspected resistance observed in the field and to elucidate the resistance mechanisms involved. The results indicated that the both resistant biotypes showed cross-resistance to diclofop-methyl (DM), pinoxaden (PN) and cycloxydim (CD) herbicides. Based on the herbicide dose that inhibited 50% of the ACCase activity (I50), the ACCase activity of the resistant biotypes was less sensitive than the S biotype to DM, CD, and PN. No differences in translocation were detected between biotypes; most of the herbicide remained in the treated leaves. The 14C-DM metabolites were identified using thin-layer chromatography. Pre-treatment with the cytochrome P450 inhibitor ABT inhibited 14C-DM metabolism in the R1 biotype, indicating that metabolism is involved in the DM resistance in the R1 biotype. DNA sequencing studies found an Ile-1781-Thr change in both resistant biotypes, conferring cross-resistance to ACCase inhibitors. In general, in the R1 biotype which showed a higher level of resistance than that of the R2 biotype, cross-resistance was observed because of mutation and DM metabolism, while in the R2 biotype, the mutation confers resistance to ACCase-inhibiting herbicides. This is the first reported evidence of the mechanisms responsible for the resistance to ACCase herbicides in P. brachystachys. These results could be useful for improved management of resistant biotypes carrying similar mutations.

Hairless Canaryseed: A Novel Cereal with Health Promoting Potential.

Glabrous canaryseeds were recently approved for human consumption as a novel cereal grain in Canada and the United States. Previously, canaryseeds were exclusively used as birdseed due to the presence of carcinogenic silica fibers; therefore the nutritional value of the seeds has been seriously overlooked. Two cultivars of glabrous canaryseeds (yellow and brown) were created from the hairy varieties. They are high in protein compared to other cereal grains, and contain high amounts of tryptophan, an amino acid normally lacking in cereals, and are gluten-free. Bioactive peptides of canaryseeds produced by in vitro gastrointestinal digestion have shown antioxidant, antidiabetic, and antihypertensive activity. The seeds contain other constituents with health promoting effects, including unsaturated fatty acids, minerals, and phytochemicals. Anti-nutritional components in the seeds are comparable to other cereal grains. Because of their beneficial health effects, canaryseeds should be regarded as a healthy food and have immense potential as a functional food and ingredient. Further research is required to determine additional bioactive peptide activity and capacity, as well as differences between the yellow and brown cultivars.

A reassessment of the genome size-invasiveness relationship in reed canarygrass (Phalaris arundinacea).

Background and Aims: Genome size is hypothesized to affect invasiveness in plants. Key evidence comes from a previous study of invasive eastern North American populations of the grass Phalaris arundinacea: invasive genotypes with smaller genomes had higher growth rates, and genome sizes were smaller in the invasive vs. native range. This study aimed to re-investigate those patterns by examining a broader range of North American populations and by employing the modern best-practice protocol for plant genome size estimation in addition to the previously used protocol. Methods: Genome sizes were measured using both internal and pseudo-internal standardization protocols for 20 invasive and nine native range accessions of P. arundinacea. After a round of vegetative propagation to reduce maternal environmental effects, growth (stem elongation) rates of these accessions were measured in the greenhouse. Key Results: Using the best-practice protocol, there was no evidence of a correlation between genome size and growth rates (P = 0.704), and no evidence for differences in genome sizes of invasive and native range accessions (P > 0.353). However, using the older genome size estimation protocol, both relationships were significant (reproducing the results of the previous study). Conclusions: Genome size reduction has not driven increased invasiveness in a broad sample of North American P. arundinacea. Further, inappropriate genome size estimation techniques can create spurious correlations between genome size and plant traits such as growth rate. Valid estimation is vital to progress in understanding the potentially widespread effects of genome size on biological processes and patterns.

Karyotype evolution in Phalaris (Poaceae): The role of reductional dysploidy, polyploidy and chromosome alteration in a wide-spread and diverse genus.

Karyotype characteristics can provide valuable information on genome evolution and speciation, in particular in taxa with varying basic chromosome numbers and ploidy levels. Due to its worldwide distribution, remarkable variability in morphological traits and the fact that ploidy change plays a key role in its evolution, the canary grass genus Phalaris (Poaceae) is an excellent study system to investigate the role of chromosomal changes in species diversification and expansion. Phalaris comprises diploid species with two basic chromosome numbers of x = 6 and 7 as well as polyploids based on x = 7. To identify distinct karyotype structures and to trace chromosome evolution within the genus, we apply fluorescence in situ hybridisation (FISH) of 5S and 45S rDNA probes in four diploid and four tetraploid Phalaris species of both basic numbers. The data agree with a dysploid reduction from x = 7 to x = 6 as the result of reciprocal translocations between three chromosomes of an ancestor with a diploid chromosome complement of 2n = 14. We recognize three different genomes in the genus: (1) the exclusively Mediterranean genome A based on x = 6, (2) the cosmopolitan genome B based on x = 7 and (3) a genome C based on x = 7 and with a distribution in the Mediterranean and the Middle East. Both auto- and allopolyploidy of genomes B and C are suggested for the formation of tetraploids. The chromosomal divergence observed in Phalaris can be explained by the occurrence of dysploidy, the emergence of three different genomes, and the chromosome rearrangements accompanied by karyotype change and polyploidization. Mapping the recognized karyotypes on the existing phylogenetic tree suggests that genomes A and C are restricted to sections Phalaris and Bulbophalaris, respectively, while genome B occurs across all taxa with x = 7.

Genotyping-by-sequencing through transcriptomics: implementation in a range of crop species with varying reproductive habits and ploidy levels.

The application of genomics in crops has the ability to significantly improve genetic gain for agriculture. Many marker-dense tools have been developed, but few have seen broad adoption in plant genomics due to issues of significant variations of genome size, levels of ploidy, single nucleotide polymorphism (SNP) frequency and reproductive habit. When combined with limited breeding activities, small research communities and scant sequence resources, the suitability of popular systems is often suboptimal and routinely fails to effectively balance cost-effectiveness and sample throughput. Genotyping-by-sequencing (GBS) encompasses a range of protocols including resequencing of the transcriptome. This study describes a skim GBS-transcriptomics (GBS-t) approach developed to be broadly applicable, cost-effective and high-throughput while still assaying a significant number of SNP loci. A range of crop species with differing levels of ploidy and degree of inbreeding/outbreeding were chosen, including perennial ryegrass, a diploid outbreeding forage grass; phalaris, a putative segmental allotetraploid outbreeding forage grass; lentil, a diploid inbreeding grain legume; and canola, an allotetraploid partially outbreeding oilseed. GBS-t was validated as a simple and largely automated, cost-effective method which generates sufficient SNPs (from 89 738 to 231 977) with acceptable levels of missing data and even genome coverage from c. 3 million sequence reads per sample. GBS-t is therefore a broadly applicable system suitable for many crops, offering advantages over other systems. The correct choice of subsequent sequence analysis software is important, and the bioinformatics process should be iterative and tailored to the specific challenges posed by ploidy variation and extent of heterozygosity.

Photo-protective mechanisms in reed canary grass to alleviate photo-inhibition of PSII on the Qinghai-Tibet Plateau.

Due to its characteristic of high biomass yield potential, there is considerable interest in cultivating Phalaris arundinacea L. cv. 'chuancaoyin No.3' (reed canary grass) on the Qinghai-Tibet Plateau where there is an abundance of alpine steppe meadow and a potential large market for animal husbandry. In this study, we 1) investigate whether reed canary grass exhibits superior productive capacity to Elymus nutans 'Aba' (E. nutans), ordinary common pasture, during the long warm days of summer at high-altitude; and 2) compare the cold tolerance between reed canary grass and E. nutans, including photosynthesis, photo-inhibition, and photo-protection. The results suggest that reed canary grass exhibits higher photosynthetic capacity compared to E. nutans at latitudes of the cool temperate zone. Meanwhile, cold-induced photo-inhibition and photo-damage at high altitudes in reed canary grass were due to both stomatal and non-stomatal limitation, and the enhancement in photo-respiration, thermal dissipation, and Mehler reaction are important processes to minimize the negative effects of high elevation and a cold environment.

Variation in sequences containing microsatellite motifs in the perennial biomass and forage grass, Phalaris arundinacea (Poaceae).

Forty three microsatellite markers were developed for further genetic characterisation of a forage and biomass grass crop, for which genomic resources are currently scarce. The microsatellite markers were developed from a normalized EST-SSR library. All of the 43 markers gave a clear banding pattern on 3% Metaphor agarose gels. Eight selected SSR markers were tested in detail for polymorphism across eleven DNA samples of large geographic distribution across Europe. The new set of 43 SSR markers will help future research to characterise the genetic structure and diversity of Phalaris arundinacea, with a potential to further understand its invasive character in North American wetlands, as well as aid in breeding work for desired biomass and forage traits. P. arundinacea is particularly valued in the northern latitude as a crop with high biomass potential, even more so on marginal lands.

Ile-1781-Leu and Asp-2078-Gly Mutations in ACCase Gene, Endow Cross-resistance to APP, CHD, and PPZ in Phalaris minor from Mexico.

Herbicides that inhibit acetyl coenzyme A carboxylase (ACCase) are commonly used in Mexico to control weedy grasses such as little seed canarygrass (Phalaris minor). These herbicides are classified into three major families (ariloxyphenoxypropionates (APP), cyclohexanodiones (CHD), and, recently, phenylpyrazolines (PPZ)). In this work, the resistance to ACCase (APP, CHD, and PPZ) inhibiting herbicides was studied in a biotype of Phalaris minor (P. minor) from Mexico, by carrying out bioassays at the whole-plant level and investigating the mechanism behind this resistance. Dose-response and ACCase in vitro activity assays showed cross-resistance to all ACCase herbicides used. There was no difference in the absorption, translocation, and metabolism of the (14)C-diclofop-methyl between the R and S biotypes. The PCR generated CT domain fragments of ACCase from the R biotype and an S reference were sequenced and compared. The Ile-1781-Leu and Asp-2078-Gly point mutations were identified. These mutations could explain the loss of affinity for ACCase by the ACCase-inhibing herbicides. This is the first report showing that this substitution confers resistance to APP, CHD, and PPZ herbicides in P. minor from Mexico. The mutations have been described previously only in a few cases; however, this is the first study reporting on a pattern of cross-resistance with these mutations in P. minor. The findings could be useful for better management of resistant biotypes carrying similar mutations.

Ecological niche differentiation of polyploidization is not supported by environmental differences among species in a cosmopolitan grass genus.

UNLABELLED: • PREMISE OF THE STUDY: Polyploidization frequently results in the creation of new plant species, the establishment of which is thought to often be facilitated by ecological niche differentiation from the diploid species. We tested this hypothesis using the cosmopolitan grass genus Phalaris (Poaceae), consisting of 19 species that range from diploid to tetraploid to hexaploid. Specifically, we tested whether (1) polyploids occupy more extreme environments and/or (2) have broader niche breadths and/or (3) whether the polyploid species' distributions indicate a niche shift from diploid species.• METHODS: We employed a bootstrapping approach using distribution data for each species and eight environmental variables to investigate differences between species in the means, extremes, and breadths of each environmental variable. We used a kernel smoothing technique to quantify niche overlap between species.• KEY RESULTS: Although we found some support for the three hypotheses for a few diploid-polyploid pairs and for specific environmental variables, none of these hypotheses were generally supported.• CONCLUSIONS: Our results suggest that these commonly held hypotheses about the effects of polyploidization on ecological distributions are not universally applicable. Correlative biogeographic studies like ours provide a necessary first step for suggesting specific hypotheses that require experimental verification. A combination of genetic, physiological, and ecological studies will be required to achieve a better understanding of the role of polyploidization in niche evolution.

Molecular genotyping of herbicide resistance in P. minor: ACCase resistance.

Little seed canary grass (Phalaris minor Retz.) populations resistant to herbicides that inhibit acetyl-CoA carboxylase (ACCase) represent an increasingly important weed control problem in northern India. The objective of this study was to develop DNA-based markers to differentiate herbicide-resistant and herbicide-susceptible population of P. minor. Primers were designed to amplify the conserved region carrying two reported mutations Trp2027 to Cys and Ile2041 to Asn conferring ACCase inhibitor resistance in several grass weeds and subjected to single-strand conformational polymorphism (SSCP) to detect the mutations. Five distinctive electrophoretic patterns on non-denaturing PAGE were observed, and four patterns were found to be associated with ACCase herbicide resistance in P. minor. The PCR-SSCP test developed in this study confirmed 17 resistant populations to contain mutations in CT domain of ACCase gene. This is the first report of rapid and easy molecular diagnosis of ACCase herbicide-resistant and herbicide-sensitive population of P. minor through PCR-SSCP analysis.

Isolation of the P5CS gene from reed canary grass and its expression under salt stress.

Reed canary grass (RCG) is a perennial grass traditionally cultivated for forage. It is also used as fuel to produce energy in Finland and Sweden, and other countries have expressed interest in the cultivation of RCG. In China, arable land is limited. Salinity is considered to be a major factor limiting plant crop development and productivity. To boost biofuel production of RCG and extend its range in saline soil, we seek to improve its salt tolerance. Proline acts as an osmolyte that accumulates when plants are subjected to abiotic stress. P5CS plays a crucial role in proline biosynthesis. We isolated a P5CS gene from RCG, designated B231P5CS (GenBank accession No. JQ622685). B231P5CS is a fragment (971 bp) that encodes a 323-amino acid polypeptide. We also cloned an actin gene fragment from RCG as a reference gene in expression analysis of B231P5CS gene. Expression analysis revealed that B231P5CS transcripts were upregulated in leaves after treatment with salt (200 mM NaCl) and that transcript levels of B231P5CS reached a maximum 12 h after exposure, which was 14.69 times the level in control plants. The trends of expression were exactly opposite in roots; transcripts were downregulated after salt treatment. Proline concentration increased in leaves after stress. In contrast, proline content of roots decreased up to 3.6-fold relative to controls. Changes in proline concentration after stress were correlated with B231P5CS expression. Our results suggest that B231P5CS is a stress-inducible gene and plays a non-redundant role in plant development. This gene may be used to improve stress tolerance of RGC and other bioenergy feedstock.

Canary grasses (Phalaris, Poaceae): biogeography, molecular dating and the role of floret structure in dispersal.

Canary grasses (Phalaris, Poaceae) include 21 species, widely spread throughout the temperate and subtropical regions of the world with two centres of diversity: the Mediterranean Basin and western North America. The genus contains annual and perennial, endemic, cosmopolitan, wild, and invasive species with diploid, tetraploid and hexaploid cytotypes. As such, Phalaris presents an ideal platform to study diversification via historic hybridization and polyploidy events, and geographical dispersal in grasses. We present the first empirical phylogeographic study for Phalaris testing current, intuitive hypotheses on the centres of origin, historic dispersal events and diversification within a geological timeframe. Bayesian methods (beast, version 1.6.2) were used to establish divergence dates, and dispersal-vicariance analyses (rasp, version 2.1b) were implemented for ancestral node reconstructions. Our phylogeographic results indicate that the genus emerged during the Miocene epoch [20.6-8.4 Ma (million years ago)] in the Mediterranean basin followed by dispersal and vicariance events to Africa, Asia and the Americas. We propose that a diploid ancestor of P. arundinacea migrated to western North America via the Bering Strait, where further diversification emerged in the New World. It appears that polyploidy played a major role in the evolution of the genus in the Old World, while diversification in the New World followed a primarily diploid pathway. Dispersal to various parts of the Americas followed different routes. Fertile florets with hairy protruding sterile lemmas showed significant correlation with wider geographical distribution.

Comparative exomics of Phalaris cultivars under salt stress.

BACKGROUND: Reed canary grass (Phalaris arundinacea) is an economically important forage and bioenergy grass of the temperate regions of the world. Despite its economic importance, it is lacking in public genomic data. We explore comparative exomics of the grass cultivars in the context of response to salt exposure. The limited data set poses challenges to the computational pipeline. METHODS: As a prerequisite for the comparative study, we generate the Phalaris reference transcriptome sequence, one of the first steps in addressing the issue of paucity of processed genomic data in this species. In addition, the differential expression (DE) and active-but-stable genes for salt stress conditions were analyzed by a novel method that was experimentally verified on human RNA-seq data. For the comparative exomics, we focus on the DE and stable genic regions, with respect to salt stress, of the genome. RESULTS AND CONCLUSIONS: In our comparative study, we find that phylogeny of the DE and stable genic regions of the Phalaris cultivars are distinct. At the same time we find the phylogeny of the entire expressed reference transcriptome matches the phylogeny of only the stable genes. Thus the behavior of the different cultivars is distinguished by the salt stress response. This is also reflected in the genomic distinctions in the DE genic regions. These observations have important implications in the choice of cultivars, and their breeding, for bio-energy fuels. Further, we identified genes that are representative of DE under salt stress and could provide vital clues in our understanding of the stress handling mechanisms in general.

Mechanism of isoproturon resistance in Phalaris minor: in silico design, synthesis and testing of some novel herbicides for regaining sensitivity.

Isoproturon, 3-p-cumenyl-1 dimethylurea was the only herbicide controlling Phalaris minor, a major weed growing in wheat fields till the early 1980s. Since it has acquired resistance against isoproturon, like other substituted urea herbicides, where the identified target site for isoproturon is in the photosynthetic apparatus at D1 protein of Photosystem-II (PS-II). Nucleotide sequence of susceptible and resistant psbA gene of P. minor has been reported to have four point mutations. During the present work D1 protein of both susceptible and resistant biotypes of P Minor has been modeled. Transmembrane segments of amino acids were predicted by comparing with the nearest homolog of bacterial D1 protein. Volume and area of active site of both susceptible and resistant biotypes has been simulated. Isoproturon was docked at the active site of both, susceptible and resistant D1 proteins. Modeling and simulation of resistance D1 protein indicates that the resistance is due to alteration in secondary structure near the binding site, resulting in loss in cavity area, volume and change in binding position, loss of hydrogen bonds, hydrophobic interaction and complete loss of hydrophobic sites. To regain sensitivity in resistant biotype new derivatives of isoproturon molecules have been proposed, synthesized and tested. Among the 17 derivatives we found that the N-methyl triazole substituted isoproturon is a potential substitute for isoproturon.

Has selection for improved agronomic traits made reed canarygrass invasive?

Plant breeders have played an essential role in improving agricultural crops, and their efforts will be critical to meet the increasing demand for cellulosic bioenergy feedstocks. However, a major concern is the potential development of novel invasive species that result from breeders' efforts to improve agronomic traits in a crop. We use reed canarygrass as a case study to evaluate the potential of plant breeding to give rise to invasive species. Reed canarygrass has been improved by breeders for use as a forage crop, but it is unclear whether breeding efforts have given rise to more vigorous populations of the species. We evaluated cultivars, European wild, and North American invader populations in upland and wetland environments to identify differences in vigor between the groups of populations. While cultivars were among the most vigorous populations in an agricultural environment (upland soils with nitrogen addition), there were no differences in above- or below-ground production between any populations in wetland environments. These results suggest that breeding has only marginally increased vigor in upland environments and that these gains are not maintained in wetland environments. Breeding focuses on selection for improvements of a specific target population of environments, and stability across a wide range of environments has proved elusive for even the most intensively bred crops. We conclude that breeding efforts are not responsible for wetland invasion by reed canarygrass and offer guidelines that will help reduce the possibility of breeding programs releasing cultivars that will become invasive.

Transgenic barley with overexpressed PTrx increases aluminum resistance in roots during germination.

A transgenic barley line (LSY-11-1-1) with overexpressed Phalaris coerulescens thioredoxin gene (PTrx) was employed to measure the growth, protein oxidation, cell viability, and antioxidase activity in barley roots during germination on the presence of 2 mmol/L AlCl(3) on filter paper. The results show that (1) compared with the non-transgenic barley, LSY-11-1-1 had enhanced root growth, although both were seriously inhibited after AlCl(3) treatment; (2) the degree of protein oxidation and loss of cell viability in roots of LSY-11-1-1 were much less than those in roots of non-transgenic barley, as reflected by lower contents of protein carbonyl and Evans blue uptakes in LSY-11-1-1; (3) activities of catalase (CAT), glutathione peroxidase (GPX), ascorbate peroxidase (APX), and glutathione reductase (GR) in LSY-11-1-1 root tips were generally higher than those in non-transgenic barley root tips, although these antioxidase activities gave a rise to different degrees in both LSY-11-1-1 and non-transgenic barley under aluminum stress. These results indicate that overexpressing PTrx could efficiently protect barley roots from oxidative injury by increasing antioxidase activity, thereby quenching ROS caused by AlCl(3) during germination. These properties raise the possibility that transgenic barley with overexpressed PTrx may be used to reduce the aluminum toxicity in acid soils.