Cytogenetic variation in somatic tissue cultures and regenerated plants of barley (Hordeum vulgare L.)

Summary Chromosome number of morphogenic and non-morphogenic calli and regenerated plants of barley were determined. Cultures were obtained from two kinds of explants, immature embryos and seedling leaves from three cultivars, Ingrid, Dissa and Golden Promise. Callus chromosome analyses were carried out during a 12 month period in a medium containing 2 mg/l of 2,4-D. Diploid cells were predominant in all cases; although in leaf-derived cultures, retraploid cells (2n=4x=28) showed a tendency to increase as time in culture increased and after more than six months in culture, diploid cells decreased to percentages of almost 70%. Aneuploid cells were generally infrequent in all cases. The source of explant has been more important than the genotype (cultivar) and the type of callus (morphogenic vs. non-morphogenic) in the chromosomal stability of cultures as time increases. From short term cultures, only 1.85% of the regenerated plants were tetraploid, the remaining were diploids. The ability of morphogenic calli to regenerate plants decreased before any significant reduction of diploid cells were observed.     

High frequency somatic embryogenesis with a pampeana-derived genotype of alfalfa (Medicago sativa L.)

Summary Somatic embryos of genotype R11 of the alfalfa variety Pampeana were produced from embryogenic calli derived from leaf sections. They were induced by an auxin shock and its development was attempted on six different media. The best condition for somatic embryo production was inducing callus on MS medium plus 10 M 2,4-D and 4,6 M KIN and transferring them, after the auxin shock, to MS with 10–20 mM NH₄ ⁺ and 30 mM proline. More than 500 somatic embryos per plate were produced. Embryos were grown to plants on MS or half strength MS media and all regenerated plants resembled the original R11 genotype. This technique could be useful in alfalfa Pampeana improvement using genetic modification.     

A comparative physiologico-genetical study of activities of some enzymes in a heterotic single cross hybrid of maize (Zea mays L.)

Summary In field trials conducted over three growth seasons I studied the differences in activities of the enzymes: polyphenoloxidase, ascorbateoxidase and peroxidase, with a heterotic single-cross hybrid of maize and its pollen-sterile analogue. Additional observations were made for differences between the single-cross hybrids and their parental lines. The samples (staminate spikelets) were taken at random just before and after tasselling. Significant differences could be established concerning the object set. Comparison of the self-pollinating line of maize with the pollen-sterile analogue revealed the down ward trend in peroxidase (P) activity with the pollen-sterile line; the upward trend in polyphenoloxidase (PPO) activity and the downward trend in ascorbateoxidase (A) activity were observed with the pollen-sterile line before tasselling. After tasselling, activity of PPO showed a downward and that of A an upward trend. The single-crospollen-sterile hybrid revealed, compared to the parental lines, an upward trend in its PPO and A activities before tasselling and a downward trend in the same enzymes after tasselling; while activity of P in this single-cross, pollen-sterile hybrid indicated the downward trend in general. Compared with the parental forms, the pollen-fertile hybrid of maize showed a trend towards lower activities of the studied enzymes for all seasons and periods under observation. Comparing results in the single-cross hybrids there was an indication of higher activities of PPO and A in the pollen-sterile analogue; whereas the same analogue revealed a lower activity of P before tasselling and higher value after tasselling.     

Screening wild plants of Capsicum annuum for resistance to pepper huasteco virus (PHV): Presence of viral DNA and differentiation among populations

Plants collected in thirteen wild populations of Capsicum annuum from Northwest Mexico were tested for resistance to the pepper huasteco begomovirus (formerly subgroup III) (PHV) that is transmitted by the white fly Bemisia tabaci Genadius. Plants were inoculated using both grafting and biolistic methods. Presence of viral DNA was detected by dot blot hybridization and densitometry. Populations varied in their resistance to PHV. Plants of only two of the populations either did not develop disease symptoms or showed very light symptoms after inoculation. In some cases, symptoms appeared several days after inoculation. In plants of these populations viral DNA was detected by dot-blot hybridization assays but they appear to be a good source of resistance (symptomless) for use in breeding programmes. This revised version was published online in August 2006 with corrections to the Cover Date.     

Sunflower mutant containing high levels of palmitic acid in high oleic background

A new sunflower mutant, CAS-12, was obtained, which has both high palmitic (≈30%) and high oleic acid contents, and also a substantial amount of palmitoleic acid (≈7%). The mutant was selected after X-ray irradiation of dry seeds of the inbred line BSD-2-423, which had normal palmitic (≈3%) and high oleic (≈88%) acid levels. The increase of palmitic and palmitoleic acids occurred at the expense of the oleic acid content, which decreased to around 55% in respect to the original line. Linoleic acid content is always under 5%. Palmitic and palmitoleic acid levels were similar to those of the high palmitic mutant CAS-5 obtained in a previous programme from a low oleic line isogenic to BSD-2-423 using a similar mutagenic treatment. In that previous programme we also selected three high stearic acid mutants using chemical mutagenic treatment on the same sunflower line (RDF-1-532). We attempted to obtain mutants in other lines but were unsuccessful. The isolation of similar mutants in isogenic parental lines illustrates the importance of the genetic background in the development of specific mutants with an altered seed oil fatty acid composition. The oil of this mutant will increase the range of potential uses of sunflower oil. This revised version was published online in July 2006 with corrections to the Cover Date.     

Precision of genetic relationship estimates based on molecular markers

Genetic progress through selection is directly related to the amount of variability present in the population and the quality of genes contributed by the parents. Genetic relationships between lines were studied using DNA marker-based estimates of genetic similarity. A statistical methodology using the width of a confidence interval was developed to determine the number of probes to be surveyed and the precision in the estimation of genetic distance between pairs of cultivars. Precision was affected by type of genetic distance used, the number of cultivars, and amount of genetic diversity present in the studied group. The width of a (1-α)% confidence interval decreased as the number of RFLP fragments increased. Oat and wheat diversity studies were used to illustrate the methodology. This revised version was published online in July 2006 with corrections to the Cover Date.     

Analysis of genetic relationships among 22 European barley varieties based on two PCR markers

Two long primers of 19 (F17) and 20 (F13) nucleotides, respectively,were used in polymerase chain reactions to amplify DNA from differentcultivated barley accessions. These primers can distinguish closely relatedvarieties and, with a unique primer, all the barley accessions analysedshowed a characteristic fingerprint. Sixty per cent and 76% of thefragments generated using F13 and F17, respectively, were polymorphic.The genetic similarity values between accessions were estimated from F13and F17 data. The dendrogram and principal coordinate analysis performedwith F13 data revealed a clear separation of these varieties in accord withtheir pedigree relationships.     

Variation for agronomic and processing traits in Solanum tuberosum haploids × wild species hybrids

Summary The broad range of wild and cultivated species relatives of the commerical potato (Solanum tuberosum L.), guarantees the existence of a gene reservoir to be screened for traits that are absent or present only in low frequencies in the cultivated germplasm. Haploids (2n=2x=24) extracted from the cultivated tetraploid (2n=4x=48) germplasm cross easily with diploid (2n=2x=24) potato species and produce fertile progeny. As most haploid × species hybrids tuberize under long day conditions they can be maintained clonally and evaluated for the traits of interest. Three populations involving Tuberosum haploids × Solanum bukasovii (tub x buk), Tuberosum haploids × Solanum sparsipilum, (tub × spl) and Tuberosum haploids × Solanum berthaultii (tub x ber) clones, were planted at two locations in Wisconsin. The haploids were derived from W231, a selected clone from the University of Wisconsin Potato Breeding Program. The objectives were: to evaluate the H-S populations for agronomic and processing traits, and to determine the phenotypic associations between them. Data were recorded on haulm maturity (HM), tuber weight (TW) and tuber number (NT) per plant, specific gravity (SG), chip color after harvest (CH), and chip color after storage at 4° C and one week of reconditioning at room temperature (CH1). Results for SG and CH indicated good potential of the germplasm evaluated to introgress these traits into the commercial potatoes. For instance, the SG mean for tub x buk, tub x spl and tub x ber were 1.079, 1.086 and 1.082, respectively, and their means for CH were 4.8, 3.9 and 3.5. Chip color after storage and reconditioning was found in low frequency in the populations. Four clones in the tub x spl population, and three clones in the tub x ber population had CH1 4.0, the commercially acceptable score for this trait. Significant (p 0.01) phenotypic correlations (r) were found between HM and SG (r=–0.46 in tub x buk, r=–0.61 in tub x spl, and r=–0.34 in tub x ber), NT and TW (r=0.79 in tub x buk, r=0.88 in tub x spl, and r=0.71 in tub x ber), and TW and SG (r=0.40 in tub x buk, and r=0.36 in tub x spl). The correlation coefficients between processing traits were not significant, which may indicate the presence of separate genetic mechanisms governing the inheritance of these traits.     

Analysis of a site x year experiment with timothy polycross progeny

Summary Results from a cooperative breeding programme with timothy for the northern areas of Scandinavia are presented. The main aim of the programme was to identify genotypes for synthetic populations that are high yielding, adapted over the whole area and stable over years. A polycross comprising 12 genotypes from each of five sites within the region was formed. The subsequent 60 half-sib families and four reference varieties were then compared under sward conditions at the same sites. The trials lasted for three years and results for total dry matter yields are presented.Significant differences in yield between lines were found. There were also significant two and three factor interactions between lines, sites and years. A new two-step procedure is presented to select the genotypes to make up a synthetic variety. Firstly, a superiority measure (Q-value) is used to select a group of genotypes that are high yielding and stable over sites and years. Secondly, the GxE part of the Q-value is split into two terms. One measures adaptation to predictable environmental conditions. The other one measures stability to unpredictable changes in the environment. A plot of these two measures provides a tool for discarding unstable or poorly adapted genotypes. The statistical properties of the selection parameters are discussed.     

Using a Chlorophyll Meter to Optimize Nitrogen Fertilizer Application for Intensively-Managed Small-Grain Cereals

Most methods used to estimate N status require collection, processing and analysis of soil and/or plant tissue samples. In this study, leaf chlorophyll content of cereals, determined in the field with a portable chlorophyll meter (Minolta SPAD-502), was used to predict N availability of wheat ( Triticum aestivum L.), barley ( Hordeum vulgare L.), oats ( Avena sativa L.) and rye ( Secale cereale L.). The results indicated that accurate nitrogen (N) fertilizer recommendations based on chlorophyll content have both environmental and economic advantages. Determination of leaf chlorophyll content with a chlorophyll meter accurately indicated plant N status allowing N fertilizer requirement to be accurately determined and resulting in increased N uptake efficiency. Applying N fertilizer on this basis improved production economics and improved physical input-output ratio during grain yield formation. Using a chlorophyll meter, responsive and non-responsive categories can be determined at the stage of maximum number of florets per ear primordium (Zadoks' GS 37-41) and at pollination (i.e. pollen grains on well-developed stigmatic hairs, GS 52-58).