Extremely high cytoplasmic diversity in natural and breeding populations of Lolium (Poaceae)

Ten chloroplast microsatellite markers were used to characterise chloroplast genetic diversity at allelic and haplotypic level in 104 accessions of Lolium perenne, other Lolium species, Festuca species and x Festulolium cultivars. Furthermore, genetic relationships between the accessions and biogeographic distribution of haplotypes were investigated using a range of Nei's population genetic diversity measures and analysis of molecular variance (AMOVA). An extremely high number (511) of haplotypes was detected in 1575 individuals. Nei's gene diversity values among L. perenne accessions ranged between 0 and 0.333. Much of the L. perenne European ecotype diversity (61%), as calculated using AMOVA, could be attributed to within-population variance and this is likely caused by, and maintained by, high levels of natural and anthropogenic seed dispersal. Plastid gene pools and maternal lineages for L. perenne could be clearly identified. Evidence was found, using AMOVA, to show a likely migration route of L. perenne from Southern regions of Europe northwards.     

Allelochemical stress inhibits growth, leaf water relations, PSII photochemistry, non-photochemical fluorescence quenching, and heat energy dissipation in three C3 perennial species

In this study, the effect of two allelochemicals, benzoxazolin-2(3H)-one (BOA) and cinnamic acid (CA), on different physiological and morphological characteristics of 1-month-old C(3) plant species (Dactylis glomerata, Lolium perenne, and Rumex acetosa) was analysed. BOA inhibited the shoot length of D. glomerata, L. perenne, and R. acetosa by 49%, 19%, and 19% of the control. The root length of D. glomerata, L. perenne, and R. acetosa growing in the presence of 1.5 mM BOA and CA was decreased compared with the control. Both allelochemicals (BOA, CA) inhibited leaf osmotic potential (LOP) in L. perenne and D. glomerata. In L. perenne, F(v)/F(m) decreased after treatment with BOA (1.5 mM) while CA (1.5 mM) also significantly reduced F(v)/F(m) in L. perenne. Both allelochemicals decreased ΦPSII in D. glomerata and L. perenne within 24 h of treatment, while in R. acetosa, ΦPSII levels decreased by 72 h following treatment with BOA and CA. There was a decrease in qP and NPQ on the first, fourth, fifth, and sixth days after treatment with BOA in D. glomerata, while both allelochemicals reduced the qP level in R. acetosa. There was a gradual decrease in the fraction of light absorbed by PSII allocated to PSII photochemistry (P) in R. acetosa treated with BOA and CA. The P values in D. glomerata were reduced by both allelochemicals and the portion of absorbed photon energy that was thermally dissipated (D) in D. glomerata and L. perenne was decreased by BOA and CA. Photon energy absorbed by PSII antennae and trapped by 'closed' PSII reaction centres (E) was decreased after CA exposure in D. glomerata. BOA and CA (1.5 mM concentration) decreased the leaf protein contents in all three perennial species. This study provides new understanding of the physiological and biochemical mechanisms of action of BOA and CA in one perennial dicotyledon and two perennial grasses. The acquisition of such knowledge may ultimately provide a rational and scientific basis for the design of safe and effective herbicides.     

巨桉林草间作模式中牧草的耐阴生理适应性

研究了不同郁闭度巨桉林下鸭茅、多年生黑麦草和扁穗牛鞭草的耐阴生理适应性.结果表明:随着林分郁闭度的增加,3种牧草的光饱和点(LSP)、光补偿点(LCP)、叶绿素a/b(Chla/Chlb)、游离脯氨酸(Pro)、可溶性糖(SS)和可溶性蛋白(SP)含量呈下降趋势,表观量子效率(AQY)、叶绿素 (Chl)含量、丙二醛(MDA)含量和超氧化物歧化酶(SOD)活性有不同程度的提高;随着林分郁闭度的增加,扁穗牛鞭草的日均净光合速率(P_n)呈下降趋势,鸭茅和多年生黑麦草的P_n先增加后下降,且在林分郁闭度分别小于0.54和0.27时,二者的P_n均高于对照;3种牧草的11项生理指标与郁闭度的相关性均达到显著水平（P<0.05）;运用平均隶属函数法对3种牧草的耐阴适应性进行排序,结果为鸭茅＞多年生黑麦草＞扁穗牛鞭草.     

盐胁迫对金牌美达丽和猎狗种子萌发的影响

用不同浓度的NaCl、KCl、MgSO₄和3种盐的复合溶液胁迫金牌美达丽和猎狗种子,观察其发芽率和萌发后子叶及胚根生长情况,对其进行生态阈限分析。结果表明:金牌美达丽和猎狗种子萌发对盐生境的适应性均很强。低浓度NaCl、KCl及复合盐溶液对猎狗种子萌发有促进作用。随盐胁迫强度上升,发芽率呈逐渐下降趋势,高浓度盐明显延缓种子的初始萌发时间,抑制幼苗正常生长。高浓度盐显著降低种子发芽率,但不同盐分对种子发芽率影响不显著。MgSO₄溶液对种子发芽率没有显著抑制现象,但对金牌美达丽胚根的生长有明显抑制作用。     

Genomic in situ hybridization (GISH) reveals high chromosome pairing affinity between Lolium perenne and Festuca mairei.

Intergeneric hybridizations have been made between species of Lolium and Festuca. It has been demonstrated, largely through conventional cytogenetic analysis, that the genomes of the two genera are related, however, much information is lacking on exactly how closely related the genomes are between the two species. We applied genomic in situ hybridization (GISH) techniques to the F1 hybrids of tetraploid Festuca mairei with a genomic constitution of M1M1M2M2 and diploid Lolium perenne with a genomic constitution of LL. It was shown in the triploid hybrids (LM1M2) that the chromosomes of M1 and M2 from F. mairei could pair with each other, and it was further discovered that L chromosomes of L. perenne paired with M1 and M2 chromosomes. Our results showed that meiocytes of Lolium-Festuca are amenable to GISH analysis, and provided direct evidence for the hypothesis that the chromosomes of Lolium and Festuca may be genetically equivalent and that reciprocal mixing of the genomes may be possible.     

A demonstration of a 1:1 correspondence between chiasma frequency and recombination using a Lolium perenne/Festuca pratensis substitution

A single chromosome of the grass species Festuca pratensis has been introgressed into Lolium perenne to produce a diploid monosomic substitution line 2n = 2x = 14. The chromatin of F. pratensis and L. perenne can be distinguished by genomic in situ hybridization (GISH), and it is therefore possible to visualize the substituted F. pratensis chromosome in the L. perenne background and to study chiasma formation in a single marked bivalent. Recombination occurs freely in the F. pratensis/L. perenne bivalent, and chiasma frequency counts give a predicted map length for this bivalent of 76 cM. The substituted F. pratensis chromosome was also mapped with 104 EcoRI/Tru91 and HindIII/Tru91 amplified fragment length polymorphisms (AFLPs), generating a marker map of 81 cM. This map length is almost identical to the map length of 76 cM predicted from the chiasma frequency data. The work demonstrates a 1:1 correspondence between chiasma frequency and recombination and, in addition, the absence of chromatid interference across the Festuca and Lolium centromeres.     

Identification of perennial ryegrass (Lolium perenne (L.)) and meadow fescue (Festuca pratensis (Huds.)) candidate orthologous sequences to the rice Hd1(Se1) and barley HvCO1 CONSTANS-like genes through comparative mapping and microsynteny

Microsynteny with rice and comparative genetic mapping were used to identify candidate orthologous sequences to the rice Hd1(Se1) gene in Lolium perenne and Festuca pratensis. A F. pratensis bacterial artificial chromosome (BAC) library was screened with a marker (S2539) physically close to Hd1 in rice to identify the equivalent genomic region in F. pratensis. The BAC sequence was used to identify and map the same region in L. perenne. Predicted protein sequences for L. perenne and F. pratensis Hd1 candidates (LpHd1 and FpHd1) indicated they were CONSTANS-like zinc finger proteins with 61-62% sequence identity with rice Hd1 and 72% identity with barley HvCO1. LpHd1 and FpHd1 were physically linked in their respective genomes (< 4 kb) to marker S2539, which was mapped to L. perenne chromosome 7. The identified candidate orthologues of rice Hd1 and barley HvCO1 in L. perenne and F. pratensis map to chromosome 7, a region of the L. perenne genome which has a degree of conserved genetic synteny both with rice chromosome 6, which contains Hd1, and barley chromosome 7H, which contains HvCO1.     

The relationships between host selection and subsequent larval performance in three free-living graminivorous sawflies

1. Selection of host plants for oviposition by females of three graminivorous sawfly species, Dolerus puncticollis C. G. Thomson, Dolerus picipes Klug. and Pachynematus clitellatus Lepeletier, was investigated experimentally using three grass species, Lolium perenne L., Festuca rubra L. and wheat, Triticum aestivum L. cv. Hunter.
2. Both species of Dolerus showed significant preferences for one grass over the other two, D. puncticollis preferring L. perenne and D. picipes selecting F. rubra. P. clitellatus females used L. perenne and F. rubra equally for oviposition, but avoided wheat.
3. The results correlated closely with larval performance trials, with D. puncticollis larvae having maximal survival and growth rates on L. perenne , and D. picipes larval performance being maximized on F. rubra. P. clitellatus larvae survived well on both grasses, but had a high level of mortality on wheat.
4. P. clitellatus larval growth rates were significantly higher on L. perenne than on F . rubra . Individual P. clitellatus adult females developing from larvae reared on L. perenne reached a significantly greater dry weight than those reared on F. rubra , but there was no significant difference in weight between males reared on the two hosts. There was some evidence that this disproportionate effect of host plant quality on the weight gain of the two sexes was translated into a bias in sex ratio of the eggs laid towards a greater proportion of females on L. perenne than on F. rubra . This manipulation of sex ratio would have a potential benefit for ovipositing females through disproportionate fecundity gains for female offspring.     

Wide hybridization experiments in cereals

Summary Wide hybridization is a useful tool in plant breeding, but little is known about its possible range. For the cereals, wheat, barley and rye, this was tested with 15 different species of the Poaceae and Panicoideae. Embryo formation could be obtained with Agropyron repens, Alopecurus agrestis, Dactylis glomerata, Festuca glauca, Hordeum bulbosum, Lolium perenne, Pennisetum americanum, and Zea mays. As well, haploid as diploid embryos occurred. New embryo culture techniques should enable these embryos to grow to plants.     

Phytoremediation of Metal-Contaminated Soil in Temperate Humid Regions of British Columbia,Canada

The suitability of five plant species was studied for phytoextraction and phytostabilisation in a region with temperate maritime climate of coastal British Columbia, Canada. Pot experiments were conducted using Lolium perenne L (perennial rye grass), Festuca rubra L (creeping red fescue), Helianthus annuus L (sunflower), Poa pratensis L (Kentucky bluegrass) and Brassica napus L (rape) in soils treated with three different metal (Cu, Pb, Mn, and Zn) concentrations. The bio-metric characters of plants in soils with multiple-metal contaminations, their metal accumulation characteristics, translocation properties and metal removal were assessed at different stages of plant growth, 90 and 120 DAS (days after sowing). Lolium was found to be suitable for the phytostabilisation of Cu and Pb, Festuca for Mn and Poa for Zn. Metal removal was higher at 120 than at 90 days after sowing, and metals concentrated more in the underground tissues with less translocation to the aboveground parts. Bioconcentration factors indicate that Festuca had the highest accumulation for Cu, Helianthus for Pb and Zn and Poa for Mn.     

Genetic and physical analysis of a single Festuca pratensis chromosome segment substitution in Lolium perenne

Molecular marker analysis and genomic in situ hybridisation (GISH) were used to examine the process of chromosome segment introgression in BC2 diploid hybrids (2n=2x=14) between Lolium perenne and Festuca pratensis. Two genotypes having what appeared to be the same, single, introgressed chromosome segment of F. pratensis in the L. perenne background were crossed with diploid L. perenne to produce a recombinant series for the introgressed region. Physical and genetic analysis of this series showed that, while recombination seemed to be possible at all points along the chromosome arm, the rate of recombination varied depending on relative position: more recombination was detected in the interstitial region as compared with the centromeric or telomeric regions. The implications of these results for the use of GISH and molecular marker analysis in the measurement of linkage drag in backcross breeding programmes is discussed.     

Segregation distortion in Lolium: evidence for genetic effects

Segregation distortion (SD) is the deviation of genetic segregation ratios from their expected Mendelian fraction and is a common phenomenon found in most genetic mapping studies. In this study two segregating Lolium perenne populations were used to construct two genetic maps: an 'F(2) biomass' consisting of 360 genotypes and an 'F(1) late flowering' sibling based population consisting of 182 genotypes. Additionally two parental maps were generated for the 'F(1) late flowering' population. SD was detected and p-values for SD were calculated for each marker locus. The 'F(1) late flowering' map had only half of the extent of SD (32%) compared to the map based on the 'F(2) biomass' population (63%). Molecular marker data have been supplemented with genomic in situ hybridization (GISH) data to show non major non-recombined segments of Fescue chromosomes within the parental inbred ryegrass lines with a Festuca x Lolium pedigree. We conclude that SD in our study is more likely caused by genetic effects rather than by population structure and marker types. Two new L. perenne mapping populations including their genetic maps are introduced; one of them is the largest reported Lolium mapping population consisting of 360 individuals.     

Surveys of new and rare microfungi in the Düsseltal (North Rhine-Westphalia)--Germany

In the years 2003 and 2004 we have observed an about 70 hectare large area in the Düsseltal, the eastern part of the Neandertal in North Rhine-Westphalia. There we collected on trees, bushes and herbs and found about 150 microfungi of which some are new for Germany or the entire world. E.g.: Pseudocercospora populigena N. ALE-AGHA, U. BRAUN & G.B. FEIGE on Populus berolinensis; Vialaea insculpta (FR.) SACC. on Ilex aquifolium L.; Passalora amelopsidis (PECK.) U. BRAUN on Parthenocissus quinquefolia (L.) PLANCH.: Pleiochaeta setosa (KIRCHN.) HUGHES on Genista angelica L.; Cercospora mercurialis PASS. on Mercurialis perennis L. (new for NRW); Pleurocytospora vestita PETRAK on Ribes aureum PURSH.; Gonatobotrys simplex CORDA on Lolium perenne L.; Phomatospora berckleyi SACC. on Dactylis glomerata L. and so on. All specimens are located in the Herbarium ESS, Mycotheca Parva collection G.B. Feige & N. Ale-Agha.     

THE FRIT FLY AND ALLIED STEM-BORING DIPTERA IN WINTER WHEAT AND HOST GRASSES

Sampling for stem-boring larvae was undertaken in 1954-56 on ley plots of Lolium perenne, Dactylis glomerata, Festuca arundinacea, F. rubra, Agrostis tenuis and Poa pratensis in randomized experiments at Hurley, Berks. The samples showed infestations by dipterous larvae, mainly species of Oscinella, Meromyza and Opomyza. The larvae showed preferences for specific grass hosts. More larvae were found where plots had been grazed throughout the season than where an early hay cut had preceded grazing. The total numbers of larvae varied between o and 688 per sq.yd. in November.
The larval populations of the leys were reflected in the populations in winter wheat which followed the plough-up of the leys. Populations of up to sixty-five larvae per sq.yd., mostly species of Oscinella , were found in the wheat in March. In 1956 a heavy larval infestation was associated with the death of considerable numbers of wheat plants.     

Expression of a novel chitinase by the fungal endophyte in Poa ampla

Many wild and cultivated cool-season grass species are naturally infected with fungal endophytes of the genera Neotyphodium and Epichlo?. These associations generally are considered mutualistic with the plants benefiting from reduced herbivory and the fungi benefiting from nutrients supplied by the plants. The fungi secrete proteins that might have a role in the interspecies symbiosis. In the interaction between Poa ampla Merr. and the endophyte Neotyphodium sp., a fungal chitinase was detected in the apoplastic protein fraction. The chitinase was also the major protein secreted in culture. Sequence analysis of the chitinase revealed it has a low level of amino acid sequence identity to other fungal chitinases and one of the conserved active site residues is altered. DNA gel-blot analysis indicated the chitinase was encoded by a single gene. Expression of similar chitinases also was detected in endophyte-infected tall fescue (Festuca arundinacea Schreb.), perennial ryegrass (Lolium perenne L.) and Chewings fescue (Festuca rubra L. subsp. fallax [Thuill] Nyman). This is the first report of an endophyte chitinase expressed in the infected host grass. As a secreted hydrolytic enzyme, the chitinase might have roles in the nutrition, growth or defense of the endophyte.     

高羊茅和多年生黑麦草内生真菌的分子检测

根据GenBank上报道的内生真菌Neotyphodium coenophialum和N.lolii的Nc25基因序列,设计了2对特异性引物FM1/R1和F3/R652,建立了一套适合于从高羊茅和多年生黑麦草种子中检测内生真菌N.coenophialum和N.lolii的常规PCR和巢式PCR方法。     

Endophytic fungi in wild and cultivated grasses in Finland

We examined the occurrence of vertically via host seeds transmitted endophyte infections of 14 grass species in natural populations in Finland and totally 97 agricultural cultivars of 13 grass species. Although endophyte infections were widespread in native grass species, overall endophyte occurrence and frequencies were lower than published reports have suggested. In natural populations, 10 out of 14 grass species examined harbor fungal endophytes in their seeds. The highest species-specific mean incidences of endophyte infected plants in infected populations were found in Agrostis capillaris. Festuca arundinacea. F. ovina. F. Pratensis. F. rubra and Phleum pratense (67%, 98%, 29%., 42%. 32% and 33%, respectively). Mean incidences were < 20% in Dactylis glomerata. Deschampsia flexuosa. D. cespitosa and Elymus repens. and no infections were detected in Calamagrostis lapponica. C. epigejos. Alopecurus pratensis and Phalaris arundinacea. However, we detected a very high variation in infection incidences among natural populations and a large proportion of populations was, indeed, endophyte-free. This supports the ideas that 1) endophytic fungi provide selective advantage of infected grasses to their uninfected eonspecifics in some habitats, and/or 2) fungi are occasionally transmitted horizontally by spores. In grass cultivars, endophyte infected seeds were detected only in F. Pratensis and Lolium perenne. and endophyte frequencies were either very high or very low. Cultivars of 11 other grass species were endophyte-free.     

Turfgrass, crop, and weed hosts of Blissus occiduus (Hemiptera: Lygaeidae)

Blissus occiduus Barber is an important pest of buffalograss, Buchlo? dactyloides (Nuttall) Engelmann, turf. No-choice studies documented the susceptibility of selected turfgrasses, crops, and weeds to B. occiduus feeding. Highly to moderately susceptible grasses included buffalograss; yellow Setaria glauca (L.) and green foxtail Setaria viridis (L.); Kentucky bluegrass, Poa pratensis L.; perennial ryegrass, Lolium perenne L.; brome, Bromus spp. Leyss.; zoysiagrass, Zoysia japonica Steudel; Bermuda grass, Cynodon dactylon (L.) Pers.; sorghum, Sorghum bicolor (L.) Moench; tall fescue, Festuca arundinacea Schreb.; and barley Hordeum vulgare (L.). Slightly to nonsusceptible grasses included fine fescue, Festuca ovina hirtula L.; rye, Secale cereale L.; crabgrass Digitaria sanguinalis (L.); bentgrass, Agrostis palustris Huds.; wheat, Tritium aestivun L.; corn, Zea mays L.; fall panicum Panicum dichotomiflorum Michx.; and St. Augustinegrass, Stenotaphrum secundatum (Walt.) Kuntze. The reproductive potential of B. occiduus was also investigated on these same grasses. B. occiduus produced offspring on 15 of the 18 turfgrass, crop, and weed species evaluated. No reproduction occurred on either Bermuda grass or St. Augustinegrass, and buffalograss plants were killed by B. occiduus feeding before offspring could be produced.     

Rapd fingerprinting of diploid Lolium perenne × hexaploid Festuca arundinacea hybrid genomes

We tested the application of RAPD technology for identification of hybrid genomes originated from a maternal clone of Lolium perenne L. (2n = 2x = 14) bearing cytoplasmic male sterility, which was pollinated separately by five clones of Festuca arundinacea Schreb. cv. Barocco (2n = 6x = 42). Six classes of RAPD markers were recognized, specific to: 1) Festuca genome and inherited into F1 hybrid genomes, 2) Lolium genome inherited into F1 hybrid genomes, 3) Lolium-specific bands not found in F1 progeny, 4) Festuca-specific bands not found in F1 progeny, 5) new bands found only in F1 hybrid profiles, 6) bands common to all parental and F1 hybrid genotypes. RAPD data were shown to have full potential a) to serve as an unequivocal proof of genome recombination in perennial ryegrass × tall fescue hybrids, b) to identify hybrid genomes, c) to reveal phenetic relationships of the accessions from crossing families, d) to enhance, by fingerprinting, the selection of superior hybrid material for further breeding. RAPD data were found to be consistent with the festucoid phenotype of F1 hybrids. This revised version was published online in July 2006 with corrections to the Cover Date.