Random amplified polymorphic DNA (RAPD) markers from leaf tissue extractions are effective for discrimination of turfgrass varieties. The usefulness of RAPD markers for turfgrass variety identification can be enhanced by use of seed rather than leaf tissue for DNA extraction. To determine whether DNA extracted from turfgrass seed was suitable for amplification, DNA was extracted from bulk samples and individual seeds of bermudagrass [Cynodon dactylon (L.) Pers.], chewings fescue (Festuca rubra var. commutata Gaud.), Poa annua L., Poa supina Schrad., creeping bentgrass [Agrostis stolonifera L. var. palustrus (Huds.) Farw.], Kentucky bluegrass (Poa pratensis L.), perennial ryegrass (Lolium perenne L.), and tall fescue (Festuca arundinacea Schreb.). All samples were successfully amplified using an arbitrary primer. Amplification intensity varied among species. With an almost infinite number of arbitrary primers available, it is likely that suitable primers can be found to amplify DNA from most turfgrass species. Amplification of turfgrass seed DNA, whether bulk or individual seed, is possible and should prove more useful than amplification of leaf tissue DNA for discrimination of turfgrass varieties.
Patricia Sweeney, Robert Golembiewski and Karl Danneberger
Patricia M. Sweeney and T. Karl Danneberger
As the number of perennial ryegrass (Lolium perenne L.) cultivars increases, the development of reliable identification methods becomes more important. Randomly amplified polymorphic DNA (RAPD) markers show promise in cultivar identification. Since perennial ryegrass cultivars are composites of genotypes rather than a single genotype, finding markers that distinguish cultivars is difficult. The ideal cultivar identification procedure would use seed tissue as a DNA source and evaluate a single sample as representative of a cultivar. The objective of this research was to determine whether RAPD markers could be used to consistently distinguish bulk seed samples of perennial ryegrass cultivars. Two extraction protocols were evaluated. A quick, simple extraction resulted in the amplification of few consistent RAPD markers. The more labor-intensive extraction with hexadecyltrimethyl ammonium bromide (CTAB), however, produced more reliable RAPD markers. Eight of 11 cultivars were distinguished by using RAPD markers produced using bulk seed samples together with four of 30 primers that were screened. These results show the potential of RAPD markers to provide the turfgrass industry, breeders, and certification agencies additional options to ensure the genetic integrity of perennial ryegrass seed lots and cultivars.
Patricia M. Sweeney and T. Karl Danneberger
The usefulness of random amplified polymorphic DNA (RAPD) in characterizing two perennial ryegrass (Lolium perenne L.) synthetic cultivars, `Accolade' and `Caravelle', was tested. Two out of 10 arbitrary primers produced three RAPD markers that distinguished bulk samples of 30 seedlings. Additional fragments were apparent when DNA from individual seedlings was amplified. Amplification products from bulk samples were not simply the sum of amplification products of individual seedlings and did not reflect all the diversity within or between the cultivars. The study illustrates the need to screen individuals to accurately evaluate the genotypic composition of a synthetic cultivar or heterogeneous population.
Patricia Sweeney, Karl Danneberger, Daijun Wang and Michael McBride
Limited information is available on the performance under temperate conditions in the United States of recently released cultivars of creeping bentgrass (Agrostis stolonifera L.) with high shoot density for use on golf course putting greens. Fifteen cultivars were established in Aug. 1996 on a greens mix with high sand content to compare their seasonal weights and total nonstructural carbohydrate (TNC) contents. The cultivars were maintained at 3.1 mm height of cut. Shoot density counts were taken during Apr., July, and Oct. 1998. Root weights and nonstructural carbohydrate levels were assessed monthly from June 1997 through Nov. 1998. A cultivar group contrast between the high shoot density cultivars (`Penn A1', `Penn A2', `Penn A4', `Penn G1', `Penn G2', and `Penn G6') and the standard cultivars (`Penncross', `Crenshaw', `Southshore', `DF-1', `Procup', `Lopez', `SR1020', and `Providence') revealed that the former averaged 342.9 and 216.1 more shoots/dm2 on two of the three sampling dates. Root dry weights did not vary significantly (P ≤ 0.05) among the cultivars. Performing a contrast between new high shoot density cultivars and standard cultivars revealed greater root dry weight in the former during Mar. and May 1998. Differences (P ≤ 0.05) in TNC were observed on two of the 18 sampling dates, but no trends were evident.
Aneta K. Studzinska, David S. Gardner, James D. Metzger, David Shetlar, Robert Harriman and T. Karl Danneberger
Turf grown in shade exhibits increased stem elongation. Dwarfism could improve turfgrass quality by reducing elongation. The purpose of this study was to examine the effect of GA2-oxidase (GA2ox) overexpression on creeping bentgrass (Agrostis stolonifera L.) performance under restricted light conditions and low mowing heights. Greenhouse studies were conducted at The Ohio State University, Columbus, OH, from 1 Sept. to 31 Oct. in both 2008 and 2009. Two experimental lines, Ax6548 and Ax6549, transformed with CP4 EPSPS and PcGA2ox gene; and a nontransformed control (NTC) was subjected to four light environments: full sun, reduced red to far red light ratio (R:FR), neutral shade [reduced photosynthetic photon flux (PPF)], and canopy shade (reduced PPF and R:FR). Turf was evaluated every 10 days for color and percent coverage. GA2ox overexpression resulted in darker green color in both transgenic lines under all light treatments as compared with NTC plants. No differences in overall turfgrass coverage were noted in full sun conditions among the lines. A significant decrease in turf coverage occurred for all shade treatments regardless of line. However, Ax6549 decreased the least. Overall data indicated that GA2ox overexpression can improve quality of turfgrass under reduced light conditions.
Edward J. Nangle, David S. Gardner, James D. Metzger, John R. Street and T. Karl Danneberger
Decreased light quantity or quality affects the growth of turfgrass plants. Shade causes thinning of turfgrass stands and loss in surface quality. Plant changes include increased chlorophyll levels, lower soluble sugars, and loss of canopy cover. The objective of this research was to investigate if applications of foliar nitrogen and trinexapac-ethyl [4-(cyclopropyl-α-hydroxy-methylene)-3,5-dioxo-cyclohexane carboxylic acid ethyl ester] (TE) would result in beneficial biochemical changes in creeping bentgrass (Agrostis stolonifera L. cv. Penncross) grown in different shaded environments. Foliar applications of three nitrogen treatments, (NH2)2CO, Ca(NO3)2, or (NH4)2SO4, were made weekly at 0.43 g N/m2. Growth regulator treatments consisted of an untreated control or TE applied biweekly at an a.i. rate of 0.057 kg·ha−1. Plots were established in full sun (FS), neutral shade (NS), and deciduous shade (DS). Chlorophyll content, soluble carbohydrates, flavonoids, clipping yield, and color were measured. Nitrogen treatments caused some variation in levels of soluble carbohydrates in shaded conditions. Chlorophyll (Chl) levels varied between TE treatments, with increased levels of chlorophyll b (Chl b) found in TE-treated plots under FS. Application of TE resulted in higher flavonoid concentrations in leaf tissue in shaded conditions. Repeated applications of (NH2)2CO significantly improved color (P = 0.05). Turfgrass managers maintaining creeping bentgrass in shade may benefit from applications of TE and (NH2)2CO.