Molecular sleuthing: The invader Cynodon case

DNA amplification fingerprinting (DAF) was used to characterize different Cynodon dactylon (L.) Pers. cultivars and accessions and evaluate the genetic diversity among them. DAF analyses of some well known cultivars used in South Africa, i.e. Sahara, Tifway, Tifgreen, Harrismith, Royal Blue, Silverton Blue, Elliott, Tifdwarf, Gulf Green, Magennis, Klip, Florida, Oval, Bayview, Bermuda, X Kweek, Natal Kweek and Tifton 78, as well as 23 unknown Cynodon specimens were done. The unknown Cynodon specimens represent invaders of cultivated lands, with considerable morphological variation. The results were used to determine the genetic distances between the cultivars and unknown accessions, to identify the unknown specimens and to determine the phylogenetic affinities between the cultivars and accessions. The results obtained from the genetic distance matrices confirmed the results obtained from the phylogenetic analyses. In this study DAF analyses was successfully used to distinguish between different C. dactylon cultivars and accessions and could be used to trace the current weed back to its introduction area. This revised version was published online in August 2006 with corrections to the Cover Date.     

Identification and polymorphism of cultivars and wild ecotypes of safflower based on isozyme patterns

Summary Acid phosphatase and cathodal peroxidase isozyme patterns were used to identify nine introduced and five local cultivars of common safflower (Carthamus tinctorius L.) and seven ecotypes of wild safflower (C. oxyacantha Bieb.). Nineteen and nine bands, respectively, were readily observed for the phosphatase and peroxidase systems in the extracts from seedling shoots. Results indicated that both systems could be used jointly for complete identification of all the 21 seed sources used.Band frequencies and polymorphic indices (PI) were determined from acid phosphatase patterns using at least 92 individual seedlings of five introduced and five local cultivars and three wild ecotypes. Band frequencies ranged from complete absence in some to total presence in other populations. The mean PI for the introduced, local and wild safflowers were found to be 0.069, 0.052 and 0.058, respectively.The banding patterns of either enzyme system were similar in some instances for the cultivated and wild safflowers and there was as much variation between two cultivars as between a cultivar and a wild ecotype, indicating close genetic relationships between C. oxyacantha and C. tinctorius.     

Isoenzyme and cotyledon protein variation for identification of black beans (Phaseolus vulgaris L.) with similar seed morphology

Summary Methods developed to identify genetically diverse varieties of beans (Phaseolus vulgaris L.) were applied to closely related lines that were difficult to distinguish on the basis of seed morphology. Seedling tissues and seeds of black beans, were examined electrophoretically for isoenzyme and cotyledon proteinn protein patterns. Seven enzymes, extracted from seeds or from seedling stem, root or leaf tissues, were compared for polymorphism. Peptidase, polyphenol oxidase, phosphoglucoisomerase and glutamate oxaloacetate transaminase patterns were the same for all lines. Some differences were observed for acid phosphatase, peroxidase and esterase patterns, but complete discrimination of the six selected lines was not possible on the basis of isoenzyme patterns alone. Using polyacrylamide gel electrophoresis (PAGE) of dissociated 0.1 M acetic acid soluble proteins at pH 3.1 (acid-PAGE), or sodium dodecyl sulphate PAGE (SDS-PAGE) of residual protein extracts, all but one pair of samples in each case could be distinguished from the other samples. Using both techniques all of the lines could be identified unequivocally.     

Field performance of bermudagrass germplasm expressing somaclonal variation selected for divergent responses to fall armyworm

Fall armyworm, Spodoptera frugiperda (J.E. Smith), is the major insect pest of bermudagrass, Cynodon dactylon (L.) Pers., in the Southeastern USA. Somaclonal variation in bermudagrass has produced distinctly divergent susceptibilities to fall armyworm under controlled conditions. Plant persistence, productivity, forage quality characteristics, and response to fall armyworm were assessed in field plots at Rosepine, Louisiana from 1995 through 1999. Forage quality characteristics were assessed each year, and natural infestations of fall armyworms were monitored in 1998 and 1999. Most of the somaclonal lines were similar to their parent cultivar in forage yield and quality, and none was consistently superior. Differences in forage quality characteristics among the bermudagrass lines were detected in 1996 and 1997, with little difference among lines in 1998 and 1999. All lines assessed were infested and sustained damage from natural infestations of fall armyworm larvae. Thus despite acceptable forage productivity and quality of most lines, expression of fall armyworm resistance under these field conditions was not detected. This lack of difference under field conditions, despite distinct resistance of some lines under controlled conditions, may primarily be due to either insufficient production of compounds deterring the insects for detectable effects under field conditions or loss of expression of the somaclonally acquired traits even with vegetative reproduction. This revised version was published online in July 2006 with corrections to the Cover Date.     

Evolution of cauliflower types grown in Great Britain as indicated by the isoenzyme composition of the cauliflower curds

Summary The curds of cauliflowers (Brassica oleracea var. botrytis L.) which are representative of the European biennials, European annuals and Australian types were used to extract 12 of the enzymes involved in carbohydrate and amino acid metabolism. Each enzyme was separated into their isoenzymes using polyacrylamide gel electrophoresis. Two enzymes, acid phosphatase EC 3.1.3.2. and aspartate aminotransferase EC 2.6.1.1. were shown to have different numbers of isoenzymes depending upon which of the three main groups of cauliflower cultivars were used. The enzymes examined showed evolutionary divergence of the cauliflower types during the selection for different times of development.     

Nitrogen requirements for deficit-irrigated bermudagrass (Cynodon spp.) fairways in South Florida

Several new bermudagrass cultivars are available and commonly used on golf course fairways. However, little is known about their cultural requirements or how these cultivars perform under lower inputs regimes. A 2-year study was conducted at University of Florida, Fort Lauderdale Research and Education Center, to assess performance of four hybrid bermudagrass [Cynodon dactylon L. Pers. X Cynodon transvaalensis (Burtt-Davy)] cultivars (‘Latitude 36’, ‘Tifway 419’, ‘TifGrand’ and ‘TifTuf’), and 2 common bermudagrass (Cynodon dactylon L. Pers.) cultivars (‘Bimini’ and ‘Celebration’) irrigated at either 50% or 80% reference evapotranspiration (ET_o) and fertilised at either 0, 146, 244, or 342 kg N ha⁻¹ year⁻¹. Plots were evaluated monthly for turfgrass quality, dark green colour index (DGCI), normalised difference vegetation index (NDVI), and Chlorophyll Index, and seasonally for N tissue content. Plots watered at 80% ET_o enhanced bermudagrass quality compared to 50% ET_o, DGCI and NDVI only in two months out of 24. Latitude 36 and Celebration were the top-rated cultivars, and their quality was not affected by no N fertilisation. Conversely, TifGrand, TifTuf and Tifway that received no N fertilisation resulted in insufficient quality during the second year of the study. Insufficient quality may be linked to reduced N metabolization compared to the highest rated cultivars. Results show that new cultivars such as Latitude 36 and Bimini could be maintained at sufficient quality levels with reduced water and N inputs in South Florida.     

Variation in the photoperiodic response on several characters in soybean,Glycine max (L.) Merrill

Summary The photoperiodic response on 12 characters in 40 soybean, Glycine max (L.) Merrill, cultivars were investigated using 10-hour (short day) and 16-hour (long day) photoperiods. Seventeen cultivars showed no significant difference between photoperiods for all 12 characters. Seven cultivars showed significant difference for only one trait. Phenotypic changes in 17 cultivars may be due to factors other than photoperiod. Number of pods per plant, days to maturity, and number of nodes per plant were highly correlated with yield per plant in both photoperiods. Invariably, the cultivars which had significant differences in the numbers of flowers produced per plant between the two photoperiods also had significant differences in the numbers of pods per plant and yield. First node to flower, plant height at flowering, days to flowering, nodes at flowering, and the 100-seed weight were least influenced by the photoperiods in most of the cultivars, while the height at maturity, number of nodes at maturity, number of days to maturity, and yield were most influenced.AVRDC Journal paper 46 (78–88).     

Effectiveness of selection for plant characters of barley in simulated segregating rows

Summary Single plants of 12 barley (Hordeum vulgare L. and H. distichum L.) cultivars were randomized to form simulated segregating (mixed) rows. There were 16 mixed rows 15 cm apart, each row containing 24 plants 5 cm apart. In addition, every third row was a control row of one of the cultivars, Bonanza. Each plant was harvested separately.The cultivars differed significantly for all seven characters measured. These seven characters fell into 3 performance groups. Group I characters, seeds/head, 1000 kernel weight, days to ripe and height, showed a higher variance in the mixed rows than in the control rows, had high average heritabilities, and were consistent in performance from one row to another when measured by correlation amongst the cultivars. In Group II, yield/plant and seeds/plant, variance, heritabilities and performance consistency were all low or non-significant. Group III, heads/plant, fell between Groups I and II. These results suggested that single plants with the desired genotype could be successfully selected for characters in Group I only.The frequency with which the most desirable genotype would actually have been chosen with 12.5% selection intensity generally confirmed the above conclusions but indicated that the degree of differences among genotypes was also important in selecting the most desirable genotypes.Contribution No. NRG 77–9 from the Beaverlodge Research Station.     

Identification of broad bean cultivars based on isozyme patterns

Summary With the increasing rate of new cultivar production for different crop plants, there is great need for methods of identifying each cultivar discriminatingly. Starch gel electrophoresis was employed to study the differences between the esterase and cathodal peroxidase isozyme patterns of 40 broad bean (Vicia faba L.) cultivars. A total of 10 and 17 medium to darkly stained bands were obtained for esterase and peroxidase systems, respectively. Bands from each enzyme system could be gropuped into three zones. Bands belonging to zone 1 of esterase (E₁) and zones 2 and 3 of peroxidase (P₂ and P₃) were quite distinct, stained intensely, and were especially useful for identification purposes.The differences in banding patterns among cultivars of the same origin were as great as those of cultivars of unrelated origin. A large proportion of the cultivars could be completely differentiated using both of the isozyme systems. There were no bands present in either enzyme system which were common to all cultivars.     

Estimation of the proportion of inbred seed in Brussels sprouts hybrid seed by acid phosphate isoenzyme analysis

Summary The acid phosphatase isoenzyme patterns of extracts of seeds from Brassica oleracea subspecies and Brussels sprouts cultivars have been analysed. In view of the inhomogeneity of most seed material, the extracts must be prepared from single seeds if the patterns are to be used for taxonomic classification.The acid phosphatase isoenzymes located in the 15–20 mm region of the electropherograms proved to be controlled by one locus, which consists of 4 alleles acting without dominance. As a result, the acid phosphatase patterns of seeds of pure inbred lines are characterized by a single band in this region. Crossing of inbred lines with a different allele constitution gives hybrid seed with the related double-band pattern. No polypeptide hybridization occurs.By analysis of the acid phosphatase patterns of sufficient individual seeds in a seed sample from a new Brussels sprouts hybrid, it was possible to determine the proportion of inbred seed in the sample. The results were in good agreement with those obtained from field observations.     

Callus induction and plant regeneration from anther and inflorescence culture of Sorghum

Summary Twenty-five inbred lines, including grain and forage types from the USA and China, two hybrids, one Sorghum almum, and one Parasorghum (S. versicolor) were tested for their response to anther culture. Three nutrient media were effective in inducing anther calli from six cultivars (Xin White, TX 403-TSB, DDY Sommer Milo, TX 2779, Brawley, and Spur Federal) and one was effective for plant regeneration for one cultivar, Xin White. Averaged over media, callus induction frequency (number of calli per 100 anthers) was highest in cultivars Xin White and TX 403-TSB (6.7 and 3.9%, respectively). The means of cultivars for media C17-2 and Ms-t-z-2, 4.3 and 3.2%, respectively, were superior to that for medium 85D3-2 (0.1%). Expressed as an average of the six cultivars and three media the mean calli induction frequency was 2.6%; however, differential responses of genotype and medium were noted. Among the 10 regeneration media tested, medium MS-d-4 containing Murashige and Skoog basal components plus 2.0 mg/l indole-3-acetic acid (IAA) and 2.5 mg/l kinetin was the most effective for plant regeneration. Numbers of albino plants and calli developing only roots increased directly with callus-induction time, whereas the frequency of plant regeneration decreased. Regenerated plants had varied numbers of chromosomes in root tip cells: 10, 15, 20, 40, and 60. The 29 regenerated plants that reached maturity, however, were highly fertile and contained only 10 bivalents in pollen mother cells. Normal chromosome number and behavior for the regenerated plants suggest that induced calli originated from cells other than microspores. However, spontaneous chromosome doubling in microspore-derived haploids may occur. The appearance of albinos also implies that haploids may have been produced from anther culture.Joint contribution of the Dept. of Agronomy and USDA-ARS, Kansas Agricultural Experiment Station, Manhattan, KS 66506-5501, USA. Contribution no. 88-566-J.     

Utilization of isozyme polymorphism for cultivar identification of 45 commercial peas (Pisum sativum L.)

Forty five Pisum sativum cultivars were analysed by isozyme electrophoresis with the objective to find protein markers for exact and reproducible discrimination of individual genotypes. The combination of six enzyme systems (acid phosphatase, amylase, esterase, leucine aminopeptidase, shikimate dehydrogenase and phosphoglucomutase) with two electrophoretic techniques (NATIVE-PAGE, isoelectric focusing) and use of seed and leaf tissue enabled to identify all 45 studied cultivars. Critical factors which may affect utilization of isozyme electrophoresis for commercial applications in pea breeding and seed production and testing are discussed. This revised version was published online in July 2006 with corrections to the Cover Date.     

Identification of tomato cultivars (Lycopersicon esculentum) by polyacrylamide isoelectric focusing

Summary Isozyme analyses have been used for the definitive identification of many plant cultivars, but not for cultivated tomatoes. Six isozyme systems, namely alcohol dehydrogenase, acid phosphatase, phosphoglucomutase, esterase, phosphoglucoisomerase and 6-phosphogluconate dehydrogenase of tomato seed extracts were resolved by isoelectric focusing on polyacrylamide gels with a narrow pH gradient. Nine alcohol dehydrogenase phenotypes were distinguished which, with three acid phosphatase phenotypes, identified twelve of the seventeen cultivars. Fewer differences were found for the other isozymes. Since this method could differentiate between breeding parents and their progeny it is concluded that further investigations are warranted.Abbreviations APS acid phosphatase - ADH alcohol dehydrogenase - EST esterase - IEF isoelectric focusing - PGI phosphoglucoisomerase - PGM phosphoglucomutase - 6-PGDH 6-phosphogluconate dehydrogenase - RFLP restriction fragment length polymorphism - VOPRI Vegetable and Ornamental Plant Research Institute     

苗龄及pH值对不同基因型甜菜根际磷酸酶活性的影响

采用无菌液培法在甜菜生长的幼苗期进行了根系内源和分泌到根际的磷酸酶的影响因子研究。结果表明,甜菜内源和分泌到根际的磷酸酶可同时存在酸性、中性和碱性3种形态。随着苗龄的变化,不同品种的内源和根际的3种磷酸酶活性变化各异,但在20 d后均明显下降。生长介质中的酸碱反应对甜菜根系各种内源磷酸酶的分泌具有明显的调     

Evaluation of seedling and adult plant resistance to leaf rust in European wheat cultivars

Summary A set of 105 European wheat cultivars, comprising 68 cultivars with known seedling resistance genes and 37 cultivars that had not been tested previously, was tested for resistance to selected Australian pathotypes of P. triticina in seedling greenhouse tests and adult plant field tests. Only 4% of the cultivars were susceptible at all growth stages. Twelve cultivars lacked detectable seedling resistance to leaf rust, and among the remaining cultivars, 10 designated genes were present either singly or in combination. Lr13 was the most frequently detected gene, present in 67 cultivars, followed by the rye-derived gene Lr26, present in 19 cultivars. Other genes present were Lr1, Lr3a, Lr3ka, Lr10, Lr14a, Lr17b, Lr20 and Lr37. There was evidence for unidentified seedling resistance in addition to known resistance genes in 11 cultivars. Field tests with known pathotypes of P. triticina demonstrated that 57% of the cultivars carried adult plant resistance (APR) to P. triticina. The genetic identity of the APR is largely unknown. Genetic studies on selected cultivars with unidentified seedling resistances as well as all of those identified to carry APR are required to determine the number and inheritance of the genes involved, to determine their relationships with previously designated rust resistance genes, and to assess their potential value in breeding for resistance to leaf rust.     

Genes conferring low seedling reaction to Mexican pathotypes of Puccinia recondita f. sp. tritici,and adult-plant responses of recent wheat cultivars from the former USSR

Fifty-five spring bread wheat (Triticum aestivum L.) cultivars, mostly released between 1975 and 1991 in eight leaf rust-prone spring wheat growing regions of the former USSR, were tested in the seedling growth stage for reaction to 15 Mexican pathotypes of Puccinia recondita f. sp. tritici. In total, seven known and at least two unknown genes were identified, either singly or in combinations: Lr3 (7 cultivars), Lr10 (14), Lr13 (5), Lr14a (1), Lr16 (1), Lr23 (3); the unknown genes were identified in 14 cultivars. The first unknown gene could be either Lr9, Lr19, or Lr25; however, the second unknown gene in 9 cultivars was different from any named gene. Twelve of the 15 pathotypes are virulent for this gene, hence its use in breeding for resistance will be limited. The cultivars were also evaluated at two field locations in Mexico with two pathotypes in separate experiments. The area under the disease progress curve and the final disease rating of the cultivars indicated genetic diversity for genes conferring adult plant resistance. based on the symptoms of the leaf tip necrosis in adult plants, resistance gene Lr34 could be present in at least 20 cultivars. More than half of the cultivars carry high to moderate levels of adult plant resistance and were distributed in each region.     

Relationship among planted and harvested kernel weights and grain yield and protein percentage in winter wheat

Summary Wheat (Triticum aestivum L.) kernel weight is an important yield component and seed quality factor that appears to be declining with recent cultivar releases in the major U.S. wheat region. The objectives of experiments were to detmrmine the relationship between planted and harvested 1000-kernel weights and their effect on grain yield and grain protein percentage. Twelve popular hard red winter wheat cultivars were grown with recommended practices at 10 Kansas locations for three years. Rank correlation coefficients between kernel weights and grain yields and protein percentages were calculated.Correlations of planted and harvested kernel weights over all cultivars with grain yield were inconsistent, being positive for some locations and years and negative for other locations and years. Planted kernel weights and grain yields of individual cultivars were not related, but harvested kernel weights and grain yields were correlated positively for eight of the 12 cultivars. Harvested kernel weights and grain protein percentages frequently were correlated positively at individual locations, but always were correlated negatively for individual cumtivars. It was concluded that high stable harvested wheat kernel weight is important as a component of grain yield under a range of environments and that improving this trait would benefit all segments of the wheat industry.Contribution No. 84-395-J, Department of Agronomy, Kansas State University, Manhattan, Kansas 66506 USA.     

Evaluation of Annual Medicago for Upper Midwest Agroecosystems

Annual Medicago species (medics) have characteristics that may make them a valuable addition to agricultural systems in the Upper Midwest, USA, but few genotypes have been evaluated. The objective of this research was to identify medic germplasm exhibiting traits that are desirable for cultivars intended for multiple uses in the region. Australian annual medic cultivars and plant introductions (PI) representing seven species were evaluated for Phytophthora root rot (Phytophthora medicaginis Hansen et Maxwell) resistance, shade tolerance, growth habit, dry matter accumulation, maturity after 9–10 weeks of growth, and potato leafhopper (Empoasca fabae Harris) yellowing. Medicago polymorpha L. plant introductions PI 197346, PI 459135, and PI 283653 were resistant to Phytophthora root rot. Kelson [Medicago scutellata (L.) Mill.] and PI 419241 [Medicago tornata (L.) Mill.] were tolerant of shade, while M. polymorpha entries Santiago and SA 9032 were intolerant. In growth habit comparisons, PI 419241 and PI 368939 (M. polymorpha) grew less than 11 cm tall, while Kelson and SA 5762 (M. polymorpha) grew at least 27 cm tall. Dry matter production 10–12 weeks after emergence was greatest for PI 197339 (M. polymorpha) and Kelson. At that time, the earliest maturing entries, PI 226517 (M. polymorpha) and Sava (M. scutellata), had begun to senesce, while the latest maturing entry (PI 419241) was still vegetative. Paraponto (Medicago rugosa Desr.) was highly resistant to potato leafhopper yellowing. None of the medic entries evaluated combined all of the desirable traits for Upper Midwest cultivars, but the traits were all present within available germplasms. Plant breeders could develop adapted cultivars by combining favourable traits from several medic genotypes.     

Forage quality of genetically diverse alfalfa germplasms at four phenological growth stages

Summary Forage quality of various alfalfa (Medicago sativa L.) cultivars has been determined under different environmental conditions, and numerous trials with alfalfa have documented an inverse relationship between forage quality and maturity. Little information is available, however, regarding the comparative forage quality of the germplasm sources from which most USA cultivars were developed. We compared forage quality of these germplasm sources at four phenological stages under disease- and insect-free conditions in a greenhouse. Germplasm sources (cultivars) tested were: Indian (Sirsa #9), African (African), Peruvian (Hairy Peruvian), Flemish (DuPuits), Turkistan (Lahontan), Chilean (Kansas Common), M. varia (Grimm), and Ladak (Ladak). Four harvests were taken and forage was separated into four phenological stages: vegetative, early bud (1–3 buds-per-stem), late bud (>3 buds-per-stem), and bloom. The germplasm source X phenological stage interaction was significant for crude protein (CP) and in vitro digestible dry matter (IVDDM) concentrations. M. varia showed the least decline in IVDDM and CP with increasing maturity. M. varia had higher IVDDM than did African and Indian at late bud and bloom stages. Indian and Flemish had higher CP than did Turkistan and Peruvian at late bud and bloom stages. Alfalfa germplasm sources differ in forage quality when comparisions are made within similar stages of phenological development.Joint contribution of the Dept. of Agronomy and USDA-ARS, Kansas Agricultural Experiment Station, Manhattan, KS 66506, USA. Contribution no. 90-475-J.