Genotypic differences in wheat Al tolerance

Summary Aluminium tolerance of 83 genotypes from Croatian and Yugoslav Triticum aestivum germplasm was evaluated in nutrient solutions having Al³⁺ activities of 0, 12.5 and 25 M. Relative root length (25 M Al³⁺/0 Al) of various genotypes ranged from 2 to 97% (from very sensitive to tolerant to Al). No genotype with Al tolerance close to that of very tolerant cultivar Atlas-66 was found. Soil, climatic, fertilization, and liming effects that wheat plants giving seeds for the nutrient solution Al-tolerance screening had been subjected to during their growth cycle did not influence the Al-tolerance ranking. Significant correlation was found between screening wheat for Al tolerance in nutrient solutions and in acid Pseudogley soil amended with five rates of limestone in a greenhouse experiment. Seed protein concentration was significantly related to the Al-tolerance ranking (r² = 0.962). Such a significant correlation was not obtained in a case of rheological and other quality characteristics of seeds. Al-tolerant wheat genotypes identified in this study will be used in breeding for improved Al tolerance.Abbreviations HSD Tukey's Honestly Significant Difference - RRL-2 relative root length, in % (12.5 M Al³⁺/0 Al) - RRL-4 relative root length, in % (25 M Al³/0 Al)     

Aiuminium tolerance of durum wheat germplasm

Summary Aluminium tolerance of Macedonian durum wheat (Triticum turgidum L. conv. durum (Desf.) MacKey) germplasm was evaluated in nutrient solutions containing 0, 74 or 148 M of total Al. Relative root length (148 M Al/0 Al) of various genotypes ranged from 41 to 72% (from moderately sensitive to moderately tolerant to Al). No genotype with Al tolerance close to that of very tolerant T. aestivum cultivar Atlas-66 was found. Seed Ca concentration was positively (r=0.64, P0.05) and seed Fe concentration negatively (r=–0.71, P0.05) related to the relative root growth. Such a significant correlation was not obtained for seed concentrations of other nutrients or seed protein content.Abbreviations HSD Tukey's Honestly Significant Difference - RRL-2 relative root length, in % (74 M total Al/0 M Al) - RRL-4 relative root length, in % (148 M total Al0 M Al)     

Studies on glucosinolates in clubroot resistant selections and susceptible commercial cultivars of cabbages

Summary Heads of 59 commercial cabbage cultivars, all susceptible to clubroot disease, and of 86 individual clubroot resistant cabbage plants of various breeding selections were analysed for their composition in glucosinolates, determined by quantifying their hydrolytic breakdown products, thiocyanate, goitrin, and volatile isothiocyanates. The mean thiocynate ion content was significantly lower in the breeding selections (120 g/g dry weight) than in the commercial cultivars (204 g/g). In contrast, the mean goitrin content was significantly higher in the breeding selections (193 g/g) than in the commercial cultivars (35 g/g). Similar to goitrin, the range of volatile isothiocyanates and total glucosinolate were higher in the breeding selections, but the mean contents of each were not statistically different between selections and cultivars. Fourteen cultivars and four selections were found to be free of goitrin; three cultivars, but no breeding selection were free of volatile isothiocyanates. The breeding selections will provide germplasm for breeding new clubroot resistant and low glucosinolate cultivars.Contribution No. J-965.     

The influence of cytokinins and ionic strength of Anderson's medium on shoot establishment and proliferation of evergreen azalea

Shoot-tip explants of evergreen azalea cv. Fuchsia grown on Anderson's medium and containing different cytokinins produced the highest proliferation rate on a medium containing thidiazuron (TDZ). TDZ concentrations ranging from 0.23 to 2.3 M resulted in both good bud-break rate (4 to 5) and shoot quality (> 0.5 cm in length). Adding 2.3 M zeatin to Anderson's medium containing 0.23 or 2.3 M TDZ increased the number of axillary shoots/explant. However, increasing the zeatin concentration to 4.6 M resulted in a reduced shoot proliferation rate. A medium containing 1.15 M TDZ and 2.3 M zeatin resulted in an 18-fold increase for 'Fuchsia' and a 9-fold increase for 'Hino Crimson' after 6 weeks of culture. It was found that explants grown on a half-strength Anderson's medium with 87.6 mM sucrose generally had better shoot proliferation rate and shoot quality than at higher ionic strength.     

Pollen grain size in four ploidy levels of genusAvena

Summary Pollen grain size in many genera positively correlated with chromosome number. In this study, oat (Avena) pollen grain size was examined in diploids (2×=14, one species, for accessions), tetraploids (4×=28, five species, 20 accessions), hexaploids (6×=42, one species, eight cultivars), and in 10 octoploid (8×=56) accessions. Mature anthers ready to dehisce pollen were sampled from one to six plants per accession, and pollen grains were squeezed out of the anther with tweezers. Oat pollen grains are slightly elliptical, and the length of the major axis was found to be highly correlated with the minor axis width (r=0.94^**). Pollen grain length, 39.3 m for diploids, 41.3 m for tetraploids, 47.0 m for hexaploids, and 48.8 m for octoploids, was positively correlated (r=0.86^**) with ploidy level. No genomic or species effects appeared to influence this trait. Only tetraploidAvena vaviloviana accession PI 412767 produced two distinct class sizes of pollen grains, 99% normal (43.0 m) and 1% large (52.7 m).     

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 tissue culture technique for vegetative propagation and low temperature preservation of Beta vulgaris

Summary A tissue culture technique for clonal propagation of Beta vulgaris is described. Flower buds or flower bud clusters formed adventitious shoots on half-strength Murashige & Skoog's medium supplemented with 10 mol/l benzyladenine (BA). The shoots were multiplied by axillary bud proliferation on a medium with 1 mol/l BA and rooted on a medium with 10 mol/l indolebutyric acid. The plants were vegetative. No mutations were observed. Genotypic variation was found in shoot formation, shoot multiplication and rooting. Some genotypes showed leaf malformations which were attributed to BA. Rooted plants in culture tubes could be stored for one year at 2 or 5°C and a low light intensity.     

Diallel analysis of yield components of snap beans exposed to two temperature stress environments

Ten snap beans (Barrier, Brio, Carson, Cornell 502, CT 70, HB 1880, Hystyle, Labrador, Opus and Venture) were selected for differential temperature tolerance and used as parents in a complete diallel mating design. The 45 F₁ hybrid lines (with reciprocals) and parents were screened at 32 C day/28 C night, and in a separate experiment, 16 C day/10 C night, during reproductive development in replicated controlled environments. Variation for yield under temperature treatments was observed among parents and hybrids, with certain hybrids exceeding parental performance. Significant (P 0.0001) general combining ability (GCA), and significant (P 0.05) specific combining ability (SCA) were observed for yield components including pod number, seed number, and seeds per pod. There was evidence that pod number and seeds per pod under temperature stress are under separate genetic control. Reciprocal effects and heterosis were not significant. GCA could not be predicted from parental performance. The breeding line Cornell 502 had the highest GCA under high temperature, and the cultivar Brio had the highest GCA under low temperature. The cross Brio × Venture was high yielding in both temperature treatments. Heat tolerance and chilling tolerance were associated in certain parents and hybrids. However, performance under high and low temperature treatments was not generally correlated in the parents and hybrids, indicating that these traits should be selected separately.     

Heritable tissue culture induced variation in Zinnia marylandica

Summary Adventitious shoots of Zinnia marylandica, an amphidiploid with limited genetic segregation, were regenerated from cotyledonary tissue on Murashige-Skoog (MS) media containing 0.2 or 22.2 M thidiazuron (TDZ) and grown through flowering. Fisher's Test for Equal Variance indicated tissue culture induced plants had more variation than seed-derived control plants. Twelve of 149 (8%) plants derived from 0.2 M TDZ and three of 23 (13%) plants from 22.2 M TDZ had variant characters. Aberrant characteristics in self-pollinated variants included plant height, fertility, flower color and morphology, and were sexually transmitted, indicating genetic change had occurred. Aberrant characteristics not observed in regenerated plants arose in progeny.Abbreviation TDZ thidiazuron     

Improved frost tolerance and winter survival in winter barley (<Emphasis Type="Italic">Hordeum vulgare</Emphasis> L.) by <Emphasis Type="Italic">in vitro</Emphasis> selection of proline overaccumulating lines

Embryogenic calli derived from anther cultures of the two-rowed winter barley cultivar Igri were plated on solid L3 medium containing the proline analogue hydroxyproline (Hyp), 10–20 mmol l^–1. Exposure to Hyp caused severe degeneration of most of the calli. Hyp resistant calli, distinguishable by their lighter colour and higher growth rate, and control calli not exposed to Hyp were plated on L3 regeneration medium. From 22,500 anthers exposed to Hyp 46 Hyp resistant regenerates were obtained, which were transferred to soil. After cultivation for 5–10 weeks at normal growth conditions they were cold hardened at 2 C under short day conditions together with control regenerates. Frost tolerance assays with segments of fully grown leaves of unhardened and cold hardened plants revealed that Hyp resistant regenerants were significantly more frost tolerant than the control regenerants. Improved frost tolerance was found also in the progenies R₁ to R₉, and genotypic segregation in the R₁ generation in a 1:2:1 ratio was indicated. Increased proline content was observed in the R₂ generation and in subsequent generations and was significantly (P 0.001) correlated with increased frost tolerance in the Hyp lines. Comparative studies of R₉ progenies from homozygous R₂ plants with the wild type Igri under field conditions in winter at three locations in Europe as well as crossing experiments confirmed the heritable improvement of frost tolerance and winter survival, respectively, in the Hyp lines. The results support the hypothesis that proline accumulation in cold acclimated winter barley plants is causally related to the acquisition of frost tolerance. Moreover, the described biotechnological procedure may be applicable in breeding programs for improved winter hardiness and possibly also for other stress tolerances.     

Interspecific hybrids of Cucumis metuliferus × C. anguria obtained through embryo culture and somatic embryogenesis

Summary Interspecific crosses between Cucumis metuliferus Naud. and C. anguria L. were obtained through embryo culture. Embryos in the rabbit-ear to advanced fluke-shaped stages were rescued 34–99 days after pollination. Plants were obtained through direct embryo culture, and through somatic embryogenesis from immature embryos. For direct embryo culture, fluke-shaped embryos were stored in sterile water in darkness for three days at 25C prior to transfer on Murashige and Skoog (MS) culture medium plus 1.0 M 6-benzylamino-purine. Multiple plants were obtained from single embryos through somatic embryogenesis of rabbit-ear stages on MS plus 10 M indole-3-acetic acid and 5 M 6-benzylamino-purine. Evidence of hybridization included leaf shape intermediate between the two parents, penduncle shape prior to fertilization which resembled the male parent, low pollen viability and isoelectric focussing of protein bands for acid phosphatase of leaf extracts.     

Effects of salicylic acid on cold preservation and cryopreservation of encapsulated embryonic axes of Persian lilac (Melia azedarach L.)

Embryonic axes of Persian lilac (Melia azedarach L.)encapsulated into calcium alginate beads with sucrose (0.75 M) and salicylicacid (0, 50 M, 200 M) were subjected to cryopreservationtechnique with dehydration and freezing in liquid nitrogen or to coldpreservation by stocking alginate beads in empty petri dishes for 4 monthsat 4 °C. In these two cases 200 M salicylic acid enhancedsignificantly the percentage of viability of encapsulated embryonic axes andthe role of salicylic acid in increasing tolerance to dehydration is discussed.     

RAPD analysis of sporting and chimerism in chrysanthemum

Summary The potential of colchicine and the microtubule depolymerizing herbicides trifluralin, oryzalin, and amiprophosmethyl (APM) for in vitro chromosome doubling during B. napus microspore culture was studied. Colchicine was administered during the first 6, 12 or 24 h of culture with 8 different concentrations up to 3 mM, and herbicides at 6 different concentrations up to 30 M for 12 h.Treatments with moderate concentrations of colchicine (3–100 M) produced a small increase in embryo production, while concentrations above 300 M were toxic. Colchicine treatment for 12 h resulted in higher embryo production than treatment for 6 and 24 h. Duration of treatment and concentration of colchicine both had a significant effect on the chromosome doubling. The highest diploidization rates (94% diploid regenerants) were seen after 24 h treatment with 1 mM colchicine.All three herbicides were similar to colchicine in terms of their effect on embryo formation and chromosome doubling comparable to the one of colchicine, but at concentrations approximately 100 times lower. APM was less toxic than trifluralin and oryzalin, but no significant difference in chromosome doubling efficiency was detected between the compounds. The 12 h treatment resulted in a maximum of approximately 65% diploid regenerants with all three herbicides, but APM may have an advantage because of its less toxic effects. Prolonged treatment with APM (20–24 h) may produce 95–100% diploid regenerants.Abbreviations APM amiprophos methyl - DMSO dimethyl sulfoxide     

Induction and verification of autotetraploids in diploid banana (Musa acuminata) by in vitro techniques

Summary Flow cytometry and stomata characteristics were used for screening ploidy levels in a large population of in vitro induced autopolyploids of the Musa acuminata breeding clone SH-3362. Culturing shoot tips in liquid medium stipplemented both with 5.0 mM colchicine for 48 hours or 30 M oryzalin (3,5-dinitro-N4,N-dipropylsulphate) for seven days, both in combination with 2% (v/v) DMSO, resulted in a high (23.1% and 29.1%) frequency of non-chimeric tetraploids in the fourth vegetative generation. Although mixoploidy persisted in subsequent cycles of vegetative propagation, tetraploids as identified by flow cytometry remained solid non-chimeric during two more cycles. These autotetraploids were propagated for field testing. A rough pre-selection of regenerated V4 plants based on their stomata characteristics resulted in a population in which only 56.2% of the plants were solid tetraploids. The somatic polyploidization system reported here can be utilised for banana breeding programmes.Abbreviations FCM Flow Cytometry     

Oil content, tocopherol composition and fatty acid patterns of the seeds of 51 Cannabis sativa L. genotypes

The oil content, the tocopherol composition, the plastochromanol-8 (P-8) content and the fatty acid composition (19 fatty acids) of the seed of 51 hemp (Cannabis sativa L.) genotypes were studied in the 2000 and 2001 seasons. The oil content of the hemp seed ranged from 26.25% (w/w) to 37.50%. Analysis of variance revealed significant effects of genotype, year and of the interaction (genotype × year) on the oil content. The oil contents of the 51 genotypes in 2000 and 2001 were correlated (r = 0.37**) and averaged 33.19 ± 1.45% in 2000 and 31.21 ± 0.96% in 2001. The -tocopherol, -tocopherol, -tocopherol, P-8- and -tocopherol contents of the 51 genotypes averaged 21.68 ± 3.19, 1.82 ± 0.49, 1.20 ± 0.40, 0.18 ± 0.07 and 0.16 ± 0.04 mg 100g^–1 of seeds, respectively (2000 and 2001 data pooled). Hierarchical clustering of the fatty acid data did not group the hemp genotypes according to their geographic origin. The -linolenic acid yield of hemp (3–30 kg ha^–1) was similar to the -linolenic acid yield of plant species that are currently used as sources of -linolenic acid (borage (19–30 kg ha^–1), evening primrose (7–30 kg ha^–1)). The linoleic acid yield of hemp (129–326 kg ha^–1) was similar to flax (102–250 kg ha^–1), but less than in sunflower (868–1320 kg ha^–1). Significant positive correlations were detected between some fatty acids and some tocopherols. Even though the average content of P-8 in hemp seeds was only 1/120^th of the average -tocopherol content, P-8 content was more closely correlated with the unsaturated fatty acid content than -tocopherol or any other tocopherol fraction. The average broad-sense heritabilities of the oil content, the antioxidants (tocopherols and P-8) and the fatty acids were 0.53, 0.14 and 0.23, respectively. The genotypes Fibrimon 56, P57, Juso 31, GB29, Beniko, P60, FxT, Félina 34, Ramo and GB18 were capable of producing the largest amounts of high quality hemp oil.     

Pedigree analysis of the origin of manganese tolerance in Canadian spring wheat (Triticum aestivum L.) cultivars

Summary Breeding wheat (Triticum aestivum L.) for tolerance to manganese (Mn) might be in some cases more feasible and economical than use of soil amendments. As part of research on the heritability of Mn tolerance, a study on the level of Mn tolerance in Canadian wheat cultivars and its probable origin was accomplished by analysis of cultivar pedigrees and drawing phylogenetic maps to discern filial relationships. Cultivar tolerance to Mn was determined by relative root weight (RRW) in solution culture in the presence of 500 M Mn. A total of 91 cultivars were screened, 76 of which were Canadian. These data, together with data from another 28 cultivars reported in the literature, were used to draw two pedigree maps, a map for Canadian cultivars only, and a map for the Mn-tolerant Canadian cultivars Norquay and Laura. Results indicated a range of tolerance to Mn among Canadian cultivars. Manganese tolerance, found in either Canadian or foreign germplasm, and of either recent or older selection or origin, seems to have originated from land races from Rio Grande do Sul, the southernmost state of Brazil. Tolerance may have been introduced into Canadian germplasm directly by the use of Brazilian cultivars as parents, or indirectly by the introduction of Mexican germplasm with Brazilian parentages. This information will help the plant breeder to develop plant breeding systems, and may also help in the study of the mechanisms for Mn tolerance in wheat.     

Breeding implications of the relationship between competitive ability and pure culture yield in self-pollinated grain crops

Summary The relative efficiencies of pedigree and bulk breeding systems for selection of high yielding genotypes are shown to be dependent on the relationship between competitive ability in mixed culture and yield in pure culture. A critical statistic is the regression coefficient, , of competitive ability (defined as the yield of a genotype in mixed culture minus its yield in pure culture) on yield in pure culture. If is less than –1, competition in mixed culture so supresses genotypes which are high yielding in pure culture conditions that they are eliminated by natural selection in early generations; pedigree selection of the lowest yielding genotypes in the F₂ and F₃ is indicated. If is greater than –1, genotypes which are high yielding in pure culture survive even when they compete less well than low yielding genotypes, and bulk breeding is indicated. An example from two sets of 100 genotypes in pure and mixed culture was analysed and the estimate of fell between 0 and –1, but because exact simulation of a segregating population was not feasible the estimate was biased. The bias tended to give estimates of that were too close to zero.Agricultural Research Council Statistics Group, Department of Applied Biology, University of Cambridge England     

Interspecific hybridization between Abelia × grandiflora ‘Francis Mason’ and A. schumannii via ovule culture

Ethiopian mustard (Brassica carinata Braun) is a potential oil crop for the Mediterranean area. The objective of this study was to develop an efficient system of mutagenesis using ultraviolet (UV) light irradiation of isolated microspores from Brassica carinata. From the survival curve based on embryo yield after irradiation of the microspores with UV light, the LD50 was estimated to be an exposure of 8 min. Total content of glucosinolates and fatty acid composition were analysed in the seeds of the doubled haploid homozygous plants with the purpose of selecting lines with modified glucosinolate and erucic acid contents. Three groups of doubled haploid lines exhibiting low and high glucosinolate contents, and high erucic acid content have been identified from a population of 270 doubled haploid lines. In eight lines, the content of glucosinolates was reduced from an average of 80.6 mol g^-1 seed to 37.5 mol g^-1 seed, whereas in four lines, the content of glucosinolates was increased up to 99.2 mol g^-1 seed. In six additional lines, the content of erucic acid was increased from 42.8% in the nontreated lines to 49.5% of the totalfatty acid composition in some of the mutant lines. All lines showed stablelevels of erucic acid in two generations, the M₂ and M₃.     

Genetic relationship between some cytoplasmic male sterility systems in sorghum

Summary Genetic analysis of cytoplasmic male sterility systems in sorghum was undertaken by evaluating a set of 25 A×B crosses and another set of 171 A×R crosses. Male steriles included diverse sources reported from U.S.A. and India. Fertility restoration in crosses was assessed by studying pollen sterility under microscope and seed setting under selfed earheads. Male fertility restorers are identified for diverse cytoplasms. Accordingly the diverse cytoplasms are grouped and listed in the increasing order of their sterility as A₁ & CK 60A A₂ A₄ M31-2A & M35-1A (A₄ tentative) A₃ & VZM2A G₁A (A₅ tentative) and consequently fertility restoration also becomes difficult in the same order for utilization in the breeding programmes. Among the non-milo cytoplasms, A₂ among exotics and maldandi (M31-2A and M35-1A) among Indian sources, can be utilised for practical exploitation in breeding programmes.     

In vitro adventitious shoot regeneration for effective maintenance of male sterile niger Guizotia abyssinica (L.f.) Cass.)

A rapid and efficient method of in vitro plant regeneration for large scale propagation of male sterile plants of niger (Guizotia abyssinica) was developed. Leaf segments from mature plants were cultured on Murashige & Skoog's basal medium (MS) supplemented with N⁶-benzyladenine (BA) and kinetin individually and in combination with low concentrations of indole-3-butyric acid (IBA); and -Naphthaleneacetic acid (NAA). Prolific direct adventitious shoot regeneration occurred on most of the media tested. The best response in terms of frequency of shoot regeneration and the number of shoots per leaf explant was observed on medium supplemented with 2.22 M BA. Transfer of shoot bud clusters to fresh medium with same composition promoted further multiplication of the shoot buds, while medium with reduced BA concentration (0.89 M) facilitated shoot elongation. Shoots that were rooted on half-strength MS medium gelled with 0.2 or 0.4% agar and supplemented with 4.9 M IBA survived with a frequency of 61.36% on transfer to ex vitrum conditions. Field evaluation of the regenerants revealed the genetic stability of the plantlets and are being used in breeding of experimental hybrids.