Selection of timothy (Phleum pratense L.) under different conditions of culture

Summary Recurrent selection for herbage weight under different conditions of competition and cutting frequency between hexaploid timothy cultivars and within their open-pollinated progenies was inefficient in producing in two generations either highly improved material as compared to the control (Climax) or any pronounced cultural ecotypes as shown by genotype x treatment interaction analyses, but some Climax and Hungarian selections were promising.Parent-offspring correlations indicated high heritability of heading and morphological characters in the first generation but not in vigor and yield which were the only characters measured in the second generation. The correlations may have been enhanced by the technique of open-pollination. A few selfings included showed very high inbreeding depression.Very early characters were useful in indicating growth types. Plant x season interactions were obvious but not tested. There were grass x legume interactions in the F₁ showing the different effects of alfalfa and white clover. A leaf marking was useful in differentiating white clover cultivars. In the F₂, spacing x cutting system interactions were significant. Light interception was significantly correlated to yields in the grass rows. Discriminant functions indicated the importance of tiller thickness for a selection index.The results are discussed in relation to experimental errors and to the possible polysomic segregation in hexaploid timothy.Published with the approval of the Director of the Ottawa Research Station, Central Experimental Farm, Ottawa. Contribution No 264.     

Use of molecular marker diversity to increase forage yield in timothy (Phleum pratense L.)

Molecular marker technologies may provide a tool to overcome the forage yield plateau in timothy (Phleum pratense L.). Therefore, a study was designed to investigate the relationship between marker‐based genetic distance (GD) estimates and yield. Genetic distances were estimated using Dice coefficients framed by 28 simple sequence repeat markers. In Experiment 1, parents with high general combining ability (GCA) from two contrasting first‐generation synthetic strains (exhibiting high and low yields) were compared. Average GDs of each strain were 0.74 (high) and 0.68 (low). These differences indicated that GD may be partially the basis for contrasting yields. In Experiment 2, GDs among 40 parents of a polycross were used for calculating general genetic distance (GGD), which is indicative of their allelic complementation. Analyses revealed a significant (P < 0.01) moderate correlation (r) between GGD and GCA for yield (r = 0.45) and a significant (P < 0.01) residual mean square for the regression of yield on GGD, suggesting that considerable non‐additive effects were associated with GCA. The results are indicative of the potential use of GD estimation for yield improvement in timothy.     

Cytogenetics of a tetraploid form of Phleum pratense L.

A tetraploid form of Phleum pratense from southwestern Europe was found to behave cytogenetically as an autotetraploid. Inheritance patterns in inbred seedlings artificially inoculated with stem rust agreed with this interpretation if chromosome and chromatid assortment are assumed.Results of cooperative investigations of the Crops Research Division, Agricultural Research Service, U.S. Department of Agriculture, and the Wisconsin Agricultural Experiment Station.Geneticist, Crops Research Division, A.R.S., U.S. Department of Agriculture, and former Graduate Assistant, Agronomy Department, Wisconsin Agricultural Experiment Station, Madison, Wisconsin.     

The Impact of Underseeding Barley (Hordeum vulgare L.) on Timothy (Phleum pratense L.)-Clover (Trifolium pratense L.; Trifolium hybridum L.) Forage Production in a Cool Maritime Climate

Growing barley (Hordeum vulgare L.) in the year of forage establishment is a common agronomic practice in marginal growing regions, but is often not recommended to growers. We examined the effect of silage barley production over an establishing timothy (Phleum pratense L.)‐clover (Trifolium pratense L.; Trifolium hybridum L.) forage sward in a 4‐year study near St. John's, Newfoundland, Canada. The experiment compared two barley varieties differing in plant height (semidwarf Chapais and Leger), three barley seeding rates and the effect of a forage understorey on forage (barley) production in the establishment year and forage (timothy‐clover) production in the subsequent year. The taller Leger yielded similar forage biomass to Chapais in the year of forage establishment, which (when planted at 375 plants m^?2) was roughly twice that of a pure‐stand timothy‐clover mix. Increasing the barley seeding rate from 125 to 375 plants m^?2 resulted in a linear increase in forage yield in the year of planting. The production of barley forage in the establishment year resulted in reduced timothy‐clover forage yield in the year following barley planted at 125 and 375 seeds m^?2, but not for barley planted at 250 seeds m^?2. Companion planting also altered forage species composition in that higher barley seeding rates resulted in 12–18 % less timothy and 2–4 % lower fibre levels in the year following planting. Barley seeded at rates of 250–375 seeds m^?2 and undersown with a timothy‐clover mixture (harvested at mid‐milk) resulted in greater forage yield of poorer quality than pure‐stand timothy‐clover in the planting year, and a barley seeding rate of 250 seeds m^?2 did not impede forage production in the subsequent year.     

Triploid and Hexaploid Regenerants from Hexaploid Timothy (Phleum pratense L.) via Anther Culture

Pollen embryos were obtained from eight genotypes and viable green plants were regenerated from four genotypes in an anther-culture experiment with 165 genotypes of Phleum pratense L. Formation of proembryos inside the cultured anthers during the first 10—12 days was significantly influenced by genotypes and by the type of nutrient media. Primary embryos developed into multiple secondary embryos before regeneration of plants. Among a total of 62 plants regenerated, only 13 were albinos. Of the green regenerants, 11 were triploid while 35 were hexaploid when DNA-content was measured by flow cytometry. Eight plants with a triploid DNA-content did possess the triploid chromosome number of 21. Triploid and hexaploid regenerants from two different parents showed simplified isozyme (GPI and PGD) banding patterns relative to that of their parents.     

Competitive Interactions between First-year Seedlings of Timothy (Phleum pratense L.) and Meadow Fescue (Festuca pratensis Huds.)

Shoot- and full-competition between first-year seedlings of meadow fescue (Festuca pratensis Huds.) and timothy (Phleum pratense L.) was studied m two separate experiments in a greenhouse. Timothy had a higher competitive ability than meadow fescue and produced more tillers, had a higher shoot weight and a larger leaf area in all strata of the canopy.
Competition for light was important up to the first harvest. The upright growth form of timothy plants during the reproductive phase enabled them to shade the unvernalized and prostrately-growing meadow fescue. Root competition may have been important afterwards. After the first harvest, tiller formation in timothy plants was high, and significantly enhanced when grown with meadow fescue. Meadow fescue produced few new tillers, and its tiller formation was depressed when competing with timothy.     

Temperature Before Cold Acclimation Affects Cold Tolerance and Photoacclimation in Timothy (Phleum pratense L.), Perennial Ryegrass (Lolium perenne L.) and Red Clover (Trifolium pratense L.)

The expected temperature rise in late summer/early autumn can change the conditions for acclimation and affect the winter survival of perennial crops. This study examined the effect of the temperature just before the onset of cold acclimation (pre‐acclimation) on freezing tolerance of timothy (Phleum pratense L.), perennial ryegrass (Lolium perenne L.) and red clover (Trifolium pratense L.) populations (both cultivars and breeding populations) adapted to either northern or southern parts of Norway. Using phytotron experiments, we studied whether increasing pre‐acclimation temperature delays growth cessation, affects photoacclimation and reduces freezing tolerance. Furthermore, we assessed whether these effects were related to the latitudinal adaptation of the plant material. The results showed that a rise in pre‐acclimation temperature decreased both cold acclimation capacity and photoacclimation in these species. This affected the freezing tolerance, which was reduced significantly more in northern‐adapted population of timothy and perennial ryegrass compared with southern‐adapted populations. Red clover was less affected by temperature changes than the grasses.     

The cytological expression and inheritance of desynapsis in a clone of diploid timothy (Phleum nodosum L.)

Summary Cytological and inheritance studies were carried out on a desynaptic clone of Phleum nodosum L. (2n=14). The clone was completely male sterile in addition to showing a medium strong degree of desynapsis. Meiotic configurations and the frequencies of various abnormalities were recorded. A significant variation in the number of bivalents at metaphase I was attributed, at least in part, to environmental factors. Cytological study of the bivalents at metaphase I in the progeny from three crosses onto the desynaptic clone revealed a close fit to a 1:1 segregation for desynaptic versus non-desynaptic plants, indicating inheritance may be controlled by a single dominant gene. The absence of male fertility in non-desynaptic plants and the observation in anther cross-sections of delayed tapetal degeneration indicated that a male sterility mechanism was also likely present in this clone.     

Impact of waterlogging and temperature on autumn growth,hardening and freezing tolerance of timothy (Phleum pratense)

Precipitation has generally increased in Norway during the last century, and climate projections indicate a further increase. The growing season has also become longer with higher temperatures, particularly in autumn. Previous studies have shown negative effects of high temperatures and, depending upon temperature conditions, contrasting effects of waterlogging on hardening capacity of timothy. We studied effects of waterlogging on seedlings of timothy (Phleum pratense, cv. Noreng) under three pre-acclimation temperatures: 3°C, 7°C, 12°C, and in autumn natural light in a phytotron at Holt, Tromsø (69°N). After temperature treatments, all plants were cold acclimated at 2°C for three weeks under continued waterlogging treatments. Freezing tolerance was determined by intact plants being frozen in pots at incremental temperature decreases in a programmable freezer. Waterlogging resulted in a higher probability of death after freezing, and a significantly reduced regrowth after three weeks at 18°C, 24 hrs light in a greenhouse. Increasing pre-acclimation temperatures also had a clear negative effect on freezing tolerance, but there was no interaction between temperatures and waterlogging. The results indicate that waterlogging may have negative implications for hardening of timothy and may contribute to reduced winter survival under the projected increase in autumn temperatures and precipitation.     

Competition Between Meadow Fescue (Festuca pratensis Huds.) and Timothy (Phleum pratense L.) at Three Levels of Nitrogen Fertilization, During Three Growing Seasons

Competition between timothy ( Phleum pratense L.) and meadow fescue ( Festucapratensis Huds.) under three levels of nitrogen was studied during three growing seasons under northern climatic conditions, 69° N latitude. Timothy initially had higher competitive abilities, particularly when nitrogen was added, but due to a higher regrowth capacity, meadow fescue was able to compete with timothy increasingly over time. The species differed in their growth patterns. Timothy had a higher proportion of reproductive tillers and a larger leaf area in the upper parts of the canopy at the first harvest, especially at high nitrogen-fertilization. Consequently, timothy shaded meadow fescue during the reproductive stage of growth. After defoliation, regrowth of timothy halted for one to two weeks. Meadow fescue, by contrast, had higher tiller numbers than timothy, a higher number of intact growing points, and resumed shoot-growth rapidly after defoliation. Due to these traits, the competitive abilities of meadow fescue increased each year relative to timothy.     

The importance of two-seeded pods in red clover (Trifolium pratense L.)

Summary The effect of selection for two-seeded pods in diploid and tetraploid populations and the effect of chromosome doubling on two-seededness were studied. The material obtained was examined to find out how important two-seeded pods are for seed set, grain weight and seed yield.In a mixoploid plant the percentage two-seeded pods in the tetraploid heads was far smaller than that in the diploid heads. Only when the character had developed relatively strongly in the diploid, did it exceed a very low value.By selection of individual plants in the best families of a diploid group of plants the percentage two-seeded pods could be raised from on average 7.4 to 30.6% as early as the following generation. Similar selection in tetraploid material with 2.5% two-seeded pods did not increase the percentage.The reduction of grain weight of seed in two-seeded pods compared with that in one-seeded pods was greater in the tetraploid plants than in the diploids.No effect of two-seededness on seed yield was found in our material. A marked effect on seed-set was observed in a diploid population (286 plants) when the percentages two-seeded pods rose about 16%.As the correlation between the calculated number of heads and the seed yield per plant is high, it may be adopted that in our material the number of heads per plant was decisive of seed yield. This will contribute to the small effect of two-seededness on the seed yield of the plant.The results of the present study showed that the production of tetraploids with a high percentage two-seeded pods will take more advantage of selection of diploid material followed by doubling of chromosomes than of direct selection in the tetraploids.Protrusion of pods in heads indicates a very high percentage two-seeded pods. This will enable relatively rapid selection for two-seeded pods.A quite small percentage two-seeded pods also occurred in two tetraploid populations that had not been selected for two-seeded pods.     

Effect of inbreeding on pseudo-self-compatibility in red clover (Trifolium pratense L.)

Summary The effect of inbreeding on pseudo-self-compatibility (PSC) was investigated in I₀, I₁, and I₂ clones of red clover, Trifolium pratense L., at 40°C. PSC was found to decrease with inbreeding. Significant differences were found among I₀ clones, among I₁ clones, and among I₂ clones for PSC. Significant differences in PSC within generations were found in two of eight I₁ clones and one of five I₂ clones.A new method of inbred line maintenance is proposed which combines the desirable features of vegetative and seed maintenance of inbred lines. Superior I₁ clones with different but homozygous S genotypes and PSC values of 8 to 10% would be vegetatively increased. Clones would be isolated under high temperature field conditions and selfed seed (PSC) would produce the I₂. Selfed seed from two different I₁ clonal lines would then be mixed and sown to produce single-cross seed. Singlecross seed obtained from different clonal sources would be mixed and sown for the production of double-cross hybrid red clover.Contribution from the Kentucky Agricultural Experiment Station. This paper (72-3-125) is published with approval of the Director, Kentucky Agricultural Experiment Station, Lexington.     

Cytology and reproductive behavior of diploid, tetraploid and hexaploid germplasm accessions of a wild forage grass: Paspalum compressifolium

Summary Fourteen germplasm accessions of Paspalum compressifolium native from southern Brazil were cytologically and embryologically analysed. The study revealed that one accession was diploid (2n=20), twelve were tetraploid (2n=40) and one was hexaploid (2n=60). This is the first report of diploid and hexaploid cytotypes for this species. Studies on microsporogenesis, megasporogenesis, and embryo sac development indicated that the diploid cytotype had regular meiotic behavior and reproduces sexually. Tetraploid cytotype usually had an important proportion of chromosomes that associated as quadrivalents during meiosis and reproduced by mean of aposporous apomixis. The hexaploid cytotype showed irregular meiotic behavior with about one third of the chromosomes associated as multivalents and reproduced by aposporous apomixis. Thus, P. compressifolium could be an agamic complex. Breeding in this complex is possible due to the presence of diploid sexuals which can be treated by colchicine and pollinated by apomicts.     

Cross compatibility between diploid and tetraploid perennial ryegrass (Lolium perenne L.)

Summary A number of crosses were made between diploid and tetraploid Lolium perenne plants in order to determine the degree of cross compatibility between the two different ploidy levels. The range of hybridization undertaken involved diploidxdiploid, and tetraploid x tetraploid control crossings, diploid x tetraploid crosses, tetraploid x diploid crosses and mixed pollinations. The seed set, the germination capacity of resultant hybrid seeds, and the chromosome constitution of derived seedlings was determined. In addition attempts were made to culture 12 day-old hybrid embryos from the diploid x tetraploid and reciprocal crosses.The crossing results obtained indicated no barrier to fertilization between diploids and tetraploids, but the irregular and poor development of the seed accompanied by much reduced germination, indicated unsatisfactory endosperm establishment of large numbers of triploids, as confirmed by the result of culturing embryos dissected from 12-day old seeds.In preliminary studies undertaken to determine the extent to which diploid and tetraploid ryegrass varieties intercross when grown in close proximity for seed production, Griffiths and Pegler (1966) observed a very low incidence of triploids amongst the progenies of diploid plants exposed to the pollen of tetraploid plants and also amongst the progenies of tetraploid material exposed to the pollen of diploid plants.In these and subsequent studies it was noted that only a very small proportion of the so-called fully formed seeds derived from diploid x tetraploid crosses did actually germinate. When, in 1964, propagules of clones of S.24 perennial ryegrass, well separated from one another, were placed in drills of the tetraploid perennial ryegrass variety Reveille, the proportion of florets which formed caryopses, and of caryopses which germinated was as follows (Table 1).The data (Table 1) clearly indicate that a considerable proportion of the S.24 florets had been fertilized by 2n pollen and had formed caryopses, but most of these had proved to be defective. As noted in the previous studies, the frequency of triploid seedlings was low, thus suggesting incompatibility between the n and 2n gametes for the production of viable zygotes.Of the total progenies classed as tetraploids in the early seedling stage, 80% proved to be ryegrass x tall fescue F₁ hybrids. These arose as a result of free crossing with     

Genetic analyses for bioactive potential in tetraploid potato (Solanum tuberosum L.)

A total of four traits related to bioactive potential of the tetraploid potato were studied in a North Carolina II cross‐experiment. The objectives of this study were to estimate genetic parameters for target bioactive compounds in a tetraploid potato population, to determine phenotypic and genotypic correlations and to calculate combining ability effects of cultivars and breeding lines included in the crossing scheme. Total monomeric anthocyanins, phenolics, carotenoids and hydrophilic antioxidant capacity were measured by spectrophotometric methods. The sources of variation female (GCA_f), male (GCA_m) and their interaction (SCA) were significant for bioactive compounds and antioxidant capacity. Narrow‐sense heritability estimates (h²) were moderate for all the traits (0.413–0.657). Both phenotypic and genotypic correlations were highly significant for phenolic compounds and antioxidant capacity. As regards combining ability, positive and negative GCA and also SCA effects have been identified for bioactive compounds and antioxidant capacity. These results indicate that hybrid breeding lines can be obtained by choosing parents with significant GCA, whose bioactive potential can also be further enhanced if the crosses have a significant SCA.     

The low down on association mapping in hexaploid wheat (Triticum aestivum L.)

The past few decades have witnessed hundreds of family-based linkage studies mapping for numerous traits but only a limited number of QTLs were actually cloned, tagged, or used for marker-assisted selection. Although providing valuable information, this conventional approach cannot be scaled up to underpin the incredible amount of phenotypic variation in the form of 266, 589 hexaploid wheat accessions maintained in public germplasm collections. Association mapping has recently emerged as an alternative and more powerful mapping approach where a natural population is surveyed to determine marker-trait associations using linkage disequilibrium (LD). After its first application for milling quality in 2006, association mapping studies in hexaploid wheat are being extended to tag yield traits, protein quality, and tolerance to biotic and abiotic stresses. Advances in genotyping technology and statistical approaches greatly accelerated the shift from conventional linkage-based mapping to LD-based association mapping. Association mapping stands out because of simultaneous utilization of a large number of ex situ-conserved natural variation due to historical recombination events accumulated over centuries.     

Estimation of outcrossing rate in landraces of tetraploid wheat (Triticum turgidum L.)

A study was undertaken to assess the amount of natural outbreeding in tetraploid wheat landraces of Ethiopian origin. The study consisted of two trials which were planted separately at two locations representing different environmental conditions for two consecutive seasons. The design employed was the randomized complete block with three replications. Two polymorphic loci, glume pubescence and awn condition were utilized as markers. The rate of outcrossing was based on the number of hybrids found in the recessive class of each marker. The overall level of outcrossing was higher in the pure stand trial than in the mixed stand trial. A higher rate of outcrossing occurred in the higher altitude area in the pure stand trial while the opposite was the case in the mixed stand trial. The average rate of outcrossing due to the marker loci was also different, with the long awn displaying more outbreeding tendency. Both locational and seasonal differences in the amount of outcrossing were observed. Generally, the study has revealed that there is a considerable amount of outbreeding in the landraces and that this genetic factor could play a role in preserving variation in the landrace populations. The level of outcrossing detected in this study could have a bearing on the maintenance of the genetic purity of diverse breeding lines being handled together in a breeding nursery and varietal purity in seed multiplication.     

The production of hexaploid Solanum x juzepczukii

The colchicine-grafting method of doubling the chromosome number of potato cuttings was used successfully for producing hexaploids from the triploid species, Solanum x juzepczukii. All the plants with hexaploid flowers were triploid/hexaploid sectorial chimeras. Seeds were obtained from crosses of the hexaploid parts with both an andigena-tuberosum hybrid and a tuberosum variety.