Cytological and Isozyme Evaluation of Tall Fescue × Italian Ryegrass Hybrids

Development of tall fescue (Festuca arundinacea Schreb., 2n = 6x - 42) × Italian rye-grass (Lolium multiflorum Lam., 2n = 2 ×= 14) hybrids would enhance efforts to improve the quality of tall fescue. Two ‘Kentucky 31’ tall fescue בLemtal Italian ryegrass hybrids were obtained via embryo rescue on MS media containing casein hydrolysate, ascorbic acid and sucrose. Chromosome pairing at metaphase I had an average of more than 12 bivalents per cell. Since Festuca-Lolium pairing can account only for seven of the paired chromosomes, intergenomic as well as interspecific chromosome pairing is indicated. There was no cytoplasmic effect on chromosome pairing. To determine if enzymes could be used as genetic markers for distinguishing hybrids from self-contaminants in crosses, zymograms of PGI, 6-PGD, MDH, GOT and ACPH were obtained from parents and hybrids using starch gel etectrophoresis. PGI, 6-PGD and MDH had fewer bands in the diploid ryegrass, as compared with the hexaploid tall fescue and the tetraploid hybrid.     

Cytogenetic and isozymic characterization of anther-panicle culture derived tall fescue aneuploids

Summary Chromosome associations of 44 tall fescue (Festuca arundinacea Schreb., 2n=6x=42) plants derived from anther-panicle culture of Kentucky 31 were evaluated to determine if new genetic stocks could be identified. Seventeen of the plants were euploids (21 II), two were monosomic (20II+1I), 22 were double monosomic (19II+2I) and three were triple monosomic (18II+3I). Zymograms were obtained for phosphoglucoisomerase (PGI), glutamate oxaloacetate transaminase (GOT), acid phosphatase (ACPH), malate dehydrogenase (MDH) and 6-phosphogluconate dehydrogenase (6-PGD). The zymograms were identical for GOT, ACPH, and 6-PGD but different patterns were found for MDH and PGI in some of the double monosomics and euploids from a different piece of callus than the majority of the plants.Contribution of the USDA-ARS in cooperation with the Kentucky Agricultural Experiment Station. Scientific Journal Series No. 86-3-61.     

Cytology and Fertility of Hybrids and Amphiploids between Aegilops caudata L. ×Triticum turgidum (L.) Thell

Interspecific hybrids and amphiploids between Aegilops caudata L. (2n = 2x = 14, CC) and Triticum turgidum (L.) Thell. conv. durum Desf M. K. (2n = 4x = 28, AABB) were produced. Such hybrids can be used to introduce desirable traits such as disease resistance into cultivated durum wheats. One of the durum parents was a ph I mutation of the cv. ‘Cappelli’ used for testing the possibility of direct introduction of alien variation into cultivated species. The amphiploids were obtained both through colchicine chromosome doubling and as natural non-reductional mciosis products. In both hybrids and amphiploids, meiotic pairing and fertility were studied. Hybrids showed varying degrees of pairing and, in addition to the one involving the ph 1 mutant, one high pairing hybrid was found (Ae. caudata× cv. ‘Capinera’). Cytological examination of microsporogenesis in amphiploids revealed a high frequency of bivalent formation. Fertility proved to be a very variable character since some of the amphiploids were almost completely sterile. The use of amphiploids in breeding programmes is discussed in relation to meiotic and fertility data.     

Comparison of the isozyme variation in tall fescue parents and their somaclones

Summary This study was conducted using the isozymes ACP-1, ADH-1, GOT-2, GOT-3, MDH, 6-PGD-1 and PGI-2 to: a) compare isozyme banding patterns of tall fescue somaclones with parents and b) correlate tissue culture-induced chromosome abnormalities with variant banding patterns. The 174 somaclones were grouped into seven categories based on their meiotic analyses and time of regeneration from culture. Differences in isozyme frequency between categories compared by chi-square tests were greatest for MDH, 6-PGD-1 and PGI-2, and least for ACP-1. The most significant differences in frequency were found between somaclones and parents. In comparisons of somaclone categories, the most different isozyme distributions were between the early vs. late regenerated somaclones. No significant differences in isozyme frequencies were found between all 42-chromosome somaclones vs. aneuploid somaclones and the three somaclone groups (42-normal, 42-abnormal, aneuploid) compared to each other. This study suggests that culture-induced isozyme variation alters the distribution of the isozyme phenotypes, but is not directly correlated with chromosome abnormalities.     

First in vivo assessment of feeding value of Festulolium hybrids derived from Festuca arundinacea var. glaucescens and selection for palatability

This paper reports on the first results of feeding value experiments, assessed with sheep, of hybrids between tetraploid Italian ryegrass (Lolium multiflorum, 2n = 4×= 28) and a progenitor of tall fescue, Festuca arundinacea var. glaucescens (2n = 4×= 28). Three hybrid populations resulting from either high, low or no selection for palatability were compared with a variety of Italian ryegrass as control and two varieties of the cultivated hexaploid (2n = 6×= 42) tall fescue, one of which had been selected for high palatability. On average, the hybrid populations were found to be as palatable as the improved tall fescue with voluntary intake (VI), in vivo digestibility of organic matter (DOM), and net energy expressed in fodder units for milk, similar to the best palatable tall fescue and to Italian ryegrass. However, selection for improved palatability among hybrids did not result in any further improvement of other traits, except VI in the hybrid selected for high palatability which significantly exceeded VI of Italian ryegrass. In conclusion, intergeneric hybridization between L. multiflorum and F. arundinacea var. glaucescens led to an immediate increase of feeding value compared with selection for palatability in tall fescue. However, to improve feeding value of hybrids further, similar selection for high palatability appeared questionable and possibly detrimental for other agronomic traits because of complicated inheritance in tetraploid L. multiflorum×F. glaucescens hybrids.     

Phenotypic polymorphism of six enzymes in the grasspea (Lathyrus sativus L.)

Summary The phenotype variation in six enzymes, 6-phosphogluconate dehydrogenase (6-PGD), malate dehydrogenase (MDH), peroxidase (PRX), isocitrate dehydrogenase (IDH), galactose dehydrogenase (GD) and glutamate oxaloacetate transaminase (GOT), was investigated using horizontal starch gel electrophoresis in 52 accessions of the grasspea, Lathyrus sativus. Phenotypic polymorphism was observed for all six enzymes. High phenotypic polymorphism (Pj) was observed for PRX and 6-PGD, while there was little polymorphism for GOT, with only two accessions showing variation. There was no correlation between phenotypic polymorphism and region of origin, or groupings of accessions made on the basis of flower colour. Tentative genetic interpretations of banding patterns are given for five of the enzyme systems. The level of apparent heterozygosity was higher than expected in this predominantly autogamous species. The level of variation in the grasspea is discussed in terms of its potential for exploitation through plant breeding.     

Genetic Diversity in Tall Fescue Ecotypes from Alabama I. Maturity, Morphological Traits, and Disease Reaction

There is a large hectarage of tall fescue in Alabama. Recognition of the deleterious effects of the endophytic fungus Acremonium coenophialum has resulted in substantial acceptance of new endophyte-free cultivars. Destruction of old pastures and the concomitant loss of potentially valuable adapted germplasm could be a valid cause for concern. The objectives of this study were to evaluate genetic variation for maturity and plant morphological traits, both among and within tall fescue cv. ‘Kentucky 31’ populations from 18 to 38 year-old pastures in Alabama. Ten populations of approximately 50 plants each were collected from old Alabama tall fescue pastures. Two pastures were sampled in each of five counties, representing the five major land use areas of the state. In 1990, spaced plants of all populations were evaluated at heading time for maturity, morphological, and disease traits at two locations in central Alabama. Endophyte infection level of the populations ranged from 2 to 100 %. Maturity was highly correlated with tiller length and flag leaf width and was used as a covariate for these traits in the analysis of variance. Leaf rust and net blotch ratings were negatively correlated. Significant variation among populations was observed for maturity and flag leaf dimensions, most variation being due to differences among source counties. Significant variation was observed within all populations for maturity and morphological traits, but only in seven populations for leaf rust. No within-population variation was detected for net blotch. The average precipitation for the month of May explained 81 % of the variation among populations for maturity. Age of the pasture sampled had a significant effect on tiller diameter (r²= 0.55), and latitude of the pasture sampled was the best individual predictor of leaf width (r²= 0.42). This study reveals that considerable genetic diversity, largely influenced by climatic conditions, exists among ‘Kentucky 31’ tall fescue ecotypes from Alabama.     

Hybrids between durum wheat and Thinopyrum junceiforme: Prospects for breeding for scab resistance

Tetraploid wheatgrass, Thinopyrum junceiforme(2n = 4x = 28; J₁J₁J₂J₂), a wild relative of wheat, is an excellent source of resistance to Fusarium head blight. Intergeneric F₁ hybrids (2n = 4x = 28; ABJ₁J₂) between durum wheat (Triticum turgidum; 2n = 4x = 28; AABB) cultivars Lloyd or Langdon and Th. junceiforme were synthesized. Most of the pairing in F₁ hybrids was between the J₁- and J₂-genome chromosomes. Some pairing occurred between wheat chromosomes and alien chromosomes, resulting in segmental exchange that was confirmed by fluorescent in situ hybridization (FISH). The F₁hybrids were largely male-sterile and were backcrossed, as the female parent, to the respective durum cultivar. Backcrosses from Lloyd × Th. junceiforme hybrids yielded fertile partial amphiploids (2n = 6x = 42; AABBJ₁J₂) as a result of functioning of unreduced female gametes of the hybrid. Lloyd proved to be a more useful durum parent than Langdon in crosses with Th. junceiforme designed to transfer scab resistance genes. Pairing in the amphiploids was characterized by preferential pairing,which resulted in bivalent formation. However, some intergeneric pairing also occurred. Several fertile hybrid derivatives were produced by further backcrossing and selfing. The introduction of alien chromatin into the durum complement was confirmed by FISH. Hybrid derivative lines had significantly lower mean infection scores (p = 0.01), the best showing 10.93% infection, whereas the parental durum cultivars had 70.34% to 89.46% infection. Hybridization with wild relatives may offer an excellent means of introducing scab resistance into durum wheat. This revised version was published online in July 2006 with corrections to the Cover Date.     

Unreduced gamete in interspecific hybrids in the Festuca/Lolium complex

Self-sterile interspecific hybrids between diploid Lolium perenne and either Festuca arundinacea or F. gigantea (both hexaploids) set some seeds when subjected to a heavy pollen cloud from the amphiploids derived from the hybrids. All the progeny were octoploid and were formed from the fertilization of unreduced gametes in the F₁ hybrid by amphiploid pollen. The significance of the results in relation to breeding efforts within the Lolium/Festuca complex is discussed.     

Tall Fescue Adaptation to Low Nitrogen Fertilization in Relation to Germplasm Type and Endophyte Infection

Efficient crop growth with low nitrogen (N) application is becoming a requirement to face the concern on excessive N emission to the environment and the increasing cost of fertilizers. This study compared six natural populations and three improved varieties of tall fescue (Festuca arundinacea Schreb.) for dry‐matter yield over 4 years under ordinary N fertilization (450 kg ha^?1 in the 4 years) and very low N fertilization (only 40 kg ha^?1 prior to sowing). Both endophyte‐infected (EI) and endophyte‐free (EF) forms of each accession were sown. On average, ordinary fertilization implied 30 % higher yield than very low fertilization and the EI germplasm outyielded the corresponding EF germplasm (+4.4 %). Mean yield of improved varieties was higher than that of natural populations under ordinary fertilization (P < 0.05), whereas the two germplasm groups did not differ under very low fertilization. Accession × N fertilization interaction was found among natural populations but not among varieties (P < 0.05). A few natural populations were top‐yielding under very low fertilization and may be exploited, possibly in combination with endophyte infection, to select tall fescue varieties with enhanced performance under limited N application.     

The addition of Hordeum chilense chromosomes to Triticum turgidum conv. durum. biochemical,karyological and morphological characterization

Summary Biochemical, karyological and morphological characterization of Triticum turgidum conv. durum/Hordeum chilense addition forms was carried out. Nine H. chilense isozyme markers, belonging to ACPH, CPX, EST, PGM, 6-PGD, GOT and MDH enzymatic systems, were used to identify the chilense chromosomes in 50 monosonic or polysomic addition forms. Several morphological traits were associated with the presence of chilense chromosome in the complement. The transmission frequencies of addition chilense chromosomes were also investigated in the offspring of various crosses.     

高羊茅光周期调控基因CONSTANS的克隆与分析

本研究以坪用型"黔草1号"高羊茅为实验材料,基于多年生黑麦草CONSTANS(CO)基因序列设计引物,利用3'RACE和5'RACE方法扩增CO基因。结果表明,从高羊茅中分离出CO基因,命名为FaCON-STANS。经序列分析,cDNA全长1557bp,编码376个氨基酸,具有完整的开放阅读框(ORF,166～1296bp),氨基端具有B-box型锌指结构域,碳端具有CCT(CO,CO-like,TOC1)结构域。进一步的同源性分析表明,Fa-CONSTANS蛋白与禾本科植物的CO蛋白亲缘关系较近,与其它植物的亲缘关系较远。     

Characterization of isozymes in Salvia columbariae

Summary Salvia columbariae is a herbaceous annual species which has potential for domestication as a new source of industrial oil. Isozyme markers provide a mean by which this process may be facilitated. Isozyme survey of field grown Salvia columbariae plants showed variation for malate dehydrogenase (MDH), phosphoglucose isomerase (PGI), and phosphogllucomutase (PGM). Selfed seed was obtained from the field and was grown in the greenhouse for segregation analyses. Electrophoretic results indicated that MDH was variable at zone 1, showing presence or absence of a band. The observed segregation ratio was not significantly different from expected ratio for Pgi-4 and Pgm-3 isozymes, indicating monogenic control of the two loci. Pgi-4 locus was heterozygous for a null allele. Cross dimerization between the allozyme Pgi-3 and Pgi-4 loci resulted in an intergenic band for this isozyme system.Abbreviations ACO aconitase - ADH alcohol dehydrogenase - DTT-DL dithiothreitol - FDH formate dehydrogenase - GOT glutamate-oxalacetate transaminase - IDH isocitric dehydrogenase - MDH malate dehydrogenase - MNR menadione reductase - 6PGD 6-phosphogluconate dehydrogenase - PGI phosphoglucose isomerase - PGM phosphoglucomutase - PVP-40 polyvinylpyrrolidone - SKDH shikimate dehydrogenase - TPI triosephosphate isomerase     

The inheritance of resistance to powdery mildew in interspecific hybrids and induced amphiploids of Zinnia elegans Jacq. and Z. angustifolia HBK

Summary Sterile interspecific hybrids and colchicine-induced amphiploids of Zinnia elegans Jacq. and Z. angustifolia HBK were examined to determine the mode of inheritance of resistance to Erysiphe cichoracearum DC ex Merat. Fertility was restored through colchicine treatment of two sterile hybrids of species reciprocal parentage which differed in ray petal response to the pathogen. Derived amphiploids were subsequently intercrossed to overcome the lack of segregation for this trait due to genetic control of pairing upon chromosome doubling. Resistance to E. cichoracearum appears to be complexly inherited in both leaves and ray florets of sterile hybrids and induced amphiploids. Two major dominant genes have been implicated in conferring resistance in ray petal tissue of derived amphiploids. Data obtained from the F₁ hybrid progeny of the intercrossed amphiploids indicate that this trait is not cytoplasmically inherited. It is speculated that the genes conferring resistance in the ray florets are acting independently from those controlling leaf resistance and that most, if not all, of the resistance genes are inherited from Z. angustifolia.Scientific Article No. A-3825, Contribution No. 6804 of the Maryland Agricultural Experiment Station.     

The Endophyte Neotyphodium coenophialum Affects Root Morphology of Tall Fescue Grown under Phosphorus Deficiency

Mechanisms involved in mineral stress tolerance of cool-season grasses infected by Neotyphodium spp. endophytes are not known. In a controlled-environment experiment, two genotypes (DN2 and DN4) of tall fescue ( Festuca arundinacea Schreb.) infected (E+) with their naturally occurring strains of N. coenophialum (Morgan-Jones and Gams) Glenn, Bacon and Hanlin, and their non-infected (E−) isolines were cultivated in nutrient solution at two phosphorus (P) levels of 31 mg P dm⁻³ (P+) and 0.31 mg P dm⁻³ (P−) for 3 weeks. Diameters of lateral roots, root hair length, and distance between root hairs were recorded using a digital image analysis system (Dage 72S CCD camera controlled by a Power MacIntosh 7200/120PC compatible computer equipped with an AG-5 frame grabber board and NIH-Image). Irrespective of tall fescue genotype and P level in nutrient solution, E+ plants had roots with a smaller diameter (16 %) than E− plants. In response to P deficiency, root diameter of E+ plants declined by 11 % and root hair length increased by 17 % when compared to E− plants. Altered root diameter and root hair length might be one of the mineral stress tolerance mechanisms in endophyte-infected tall fescue.     

Impact of Phosphorus from Dairy Manure and Commercial Fertilizer on Perennial Grass Forage Production

Increased recovery and recycling of manure phosphorus (P) by crops on dairy farms is needed to minimize environmental problems. The main objective of this study was to compare P utilization by orchardgrass (Dactylis glomerata L.) and tall fescue (Festuca arundinaceae Schreb.) from dairy manure or inorganic fertilizer. The study was conducted from 1994 to 2000 at the Cornell University Baker Farm, Willsboro, NY, on a somewhat poorly drained Kingsbury clay (very–fine, illitic, mesic Aeric Epiaqualfs). The design was a split‐plot in a randomized complete block with two manure rates (16 800 and 33 600 kg ha^?1) and one nitrogen (N) fertilizer rate (84 kg N ha^?1 at spring greenup and 56 kg N ha^?1 prior to each regrowth harvest) as the main plots and grass species as subplots replicated six times. Fertilizer P [Ca(H₂PO₄)₂] was applied to the fertilizer treatment in 1995 and 1996 at 11 kg P ha^?1 year^?1. Orchardgrass P removal averaged 21 % higher than tall fescue P removal for the spring harvest, but orchardgrass averaged 24 % lower P removal than tall fescue removal for all regrowth harvests from 1995–99. Phosphorus herbage concentration in the fertilizer treatment was in the range of 1.9–2.7 g P kg^?1 compared with 2.2–5.3 g P kg^?1 in the manure treatments. Seasonal P removal ranged from as low as 9.2 kg P ha^?1 to as high as 48.5 kg P ha^?1. Morgan extractable soil P in the top 0–0.20 m remained high through 1999, with 29.1 kg P ha^?1 at the highest manure rate in tall fescue compared with 8.4 kg P ha^?1 measured in 1993 prior to the experiment. In 2000, soil P at the highest manure rate in tall fescue dropped to 10.1 kg P ha^?1, following cessation of manure application in 1998. Intensively managed harvested orchardgrass and tall fescue have the potential to remove large quantities of manure P.     

Tall and Hard Fescue Responses to Periodic Soil Water Deficits

Tall (Festuca arundmacea Schreb.) and hard (Festuca ovina var. longifolia (L.) Koch) fescues are widely sown to stabilize disturbed soils in the cool–humid and transition climatic zones of the USA. Our objectives were to: a) quantify changes in the allocation of dry matter and growth of tall and hard fescue, forced to grow on stored soil water in sandy soil; and b) compare dry matter allocation over time between roots and shoots to evaluate the dynamics of root-associated drought avoidance mechanisms. A randomized complete block experiment with four replications and two factors (species and stress level) was conducted in a greenhouse. The main blocks consisted of two fescue species: tall fescue and hard fescue; sub-blocks contained stress levels: well-watered and stressed. Low, medium and severe stresses were imposed by withholding water in one set of pots. The types of fescue species grown significantly affected leaf area (LA), plant height (PH), water use (WU), root length (RL), longest root (LR), root area (RA), shoot (SDB) and root dry biomass (RDB), and root:shoot ratio (R/S). Stress level affected PH, WU, LR and RL at low stress; WU, LR and RA at medium stress; LA, PH, SDB, WU, RL, LR, RA and R/S at severe stress. Tall fescue had greater LA, PH, WU, RA, RL, LR, SDB, RDB and R/S than hard fescue under all treatments. Stress reduced LA, PH, SDB, WU, RA, RL, LR and R/S. Significant correlations were obtained for LA with RL, WU, LR, PH. RA, SDB, RDB; RL with WU, LR, PH. RA, SDB. RDB; WU with LR, PH, RA, SDB, RDB; LR with PH. RA, SDB, RDB; PH with RA, SDB, RDB; RA with SDB, RDB; and SDB with RDB. In conclusion, hard fescue had a shallower root system, shorter plant canopy, slower growth, and transpired less water to make it more drought tolerant. Tall fescue, with a deeper root system, longer plant canopy, faster growth, and greater water transpiration, is less drought tolerant at medium and severe stresses. Root attributes strongly correlated with shoot attributes and can be considered for breeding programs promoting drought tolerance.     

同工酶分析法鉴定百合杂种F1代

为提高育种的选择效率,及时、准确鉴别杂种F1代的真伪,采用聚丙烯酰胺凝胶电泳法,对百合11个杂交组合40个后代进行同工酶分析。结果如下：（1）鉴定的40个后代中,共有30个后代可以确定为真杂种。（2）11个杂交组合的酯酶、莽草酸脱氢酶、苹果酸脱氢酶、6-磷酸葡萄糖脱氢酶分析中,父本相较于母本存在特征性条带的杂交组合数分别为8、2、0、0。（3）组合2、3为正反交关系,并未发现父本或母本的特异性遗传。（4）同一品种不同时期,酯酶的酶谱上存在着一定差异,但其基带及特异性条带相对稳定,并未影响到杂种后代的最终判定结果。可见,同工酶技术是鉴定百合杂种的有效手段,酯酶及莽草酸脱氢酶在其中发挥着重要作用。     

Hybridizing <Emphasis Type="Italic">Brassica rapa</Emphasis> with wild crucifers <Emphasis Type="Italic">Diplotaxis erucoides</Emphasis> and <Emphasis Type="Italic">Brassica maurorum</Emphasis>

Two wide hybrids, Diplotaxis erucoides (2n = 14) × Brassica rapa (2n = 20) and B. maurorum (2n = 20) × B. rapa, were developed using the sequential ovary–ovule culture. Reciprocal crosses failed, possibly as a consequence of strong unilateral incompatibility. The F ₁ hybrids in each combination were completely male sterile and morphologically intermediate to the respective parents. DNA marker polymorphism and chromosome counts confirmed their hybrid nature. High frequency of bivalents in the F ₁ and the presence of trivalents/quadrivalents in the derived amphiploids suggested genomic duplications and homoeology of the parental genomes. Up to three homoeologous pairs between the D. erucoides (D^eD^e) and B. rapa (AA) genomes, and one between B. maurorum (B^mB^m) and B. rapa genomes were observed. Successful synthesis of the F ₁ hybrids and amphiploids of B. rapa with D. erucoides and B. maurorum, and allosyndetic chromosome pairing are expected to permit introgressions of desirable loci into the cultivated Brassica germplasm, especially for resistance to Alternaria brassicae and Albugo candida.     

Introgression of crown rust resistance from Festuca spp. into Lolium multiflorum

Crown rust, Puccinia coronata Corda, causes one of the most damaging foliar diseases in Italian ryegrass, Lolium multiflorum Lam. For introgression of crown rust resistance, highly resistant hybrids of the crosses Festuca arundinacea (2n = 6x = 42) ×L. multiflorum (2n = 4x = 28) and reciprocally, L. multiflorum (2n = 2x = 14) ×Festuca pratensis (2n = 4x = 28) and subsequently resistant recombinant individuals were used as female parents and susceptible cultivars of Italian ryegrass as male parents in three successive backcrosses. The BC₃ plants were selfed and crossed mutually. Uredospores of seven different crown rust isolates collected from plants of L. multiflorum, Lolium perenne, F. pratensis, F. arundinacea and L. multiflorum × F. pratensis hybrids were applied to identify the resistance or susceptibility of the parental species, backcrossed, selfed and intercrossed progenies. The various crown rust populations revealed a species-specific capability to infect plants of the Lolium-Festuca complex corresponding to the host species from which the spores originated. Selected BC⁹ plants, however, were found to be completely resistant to all crown rust populations tested. Successful introgression of the resistance was achieved from F. arundinacea as well as from F. pratensis. The resistance represents a dominant character, apparently based on a strong heterologous incompatibility between host and pathogen. In phenotype, bivalent formation during meiosis and in fertility, the novel germplasms are comparable with the L. multiflorum cultivars.