Somatic embryogenesis from floral tissue of cassava (Manihot esculenta Crantz)

The aim of this study was to examine the embryogenic potential of floral material of the cassava cultivar MCOL 1505. Macerated immature inflorescences were found to be highly embryogenic, with almost 78% of the explants producing somatic embryos. Somatic embryos were also produced from whole male florets and half florets although at much lower rates. No regeneration was obtained from anther, microspore or floret wall tissue. Somatic embryos derived from immature inflorescences were regenerated via organogenesis and the plants derived from this process were assessed in terms of phenotype and ploidy level. If haploid plants could be produced by this method, this would have significant implications in assisting traditional cassava breeding, as this would allow homozygosity to be reached more rapidly. In a crop such as cassava, which is highly heterozygous in nature, the use of haploids in a breeding programme could considerably shorten the time taken to produce new desirable cultivars. This is the first report on plant regeneration through somatic embryogenesis from floral tissue of cassava. This revised version was published online in July 2006 with corrections to the Cover Date.     

Effect of incubation temperature regimes and culture medium on broccoli microspore culture embryogenesis

Conditions for reliable induction of embryogenesis from isolated microspores were studied in ten genotypes of broccoli. Embryo yields were significantly increased in almost all of the broccoli genotypes by the incubation at 32.5 ^°C for 1 day, than when the standard incubation at 30 ^°C for 2 days was used. Treatments of 48 hours at 32.5 ^°C produced less than optimal results suggesting that broccoli microspores are more sensitive to high temperatures than those of B. napus. The use of the ¹/₂ NLN-13 medium yielded greater number of embryos than the standard NLN-13. The magnitude of the response to the redution of the concentration of major salts by half in the NLN medium varied with the different genotypes. High embryogenic broccoli cultivars, such as ‘Shogun’, ‘SDB9’, and ‘Green Valiant’, presented a better response to the reduction of the concentration of major salts by half in NLN-13. Reduction never produced a detrimental effect on embryo yield and seems not to have any effect in the subsequent development of embryos in plants. This revised version was published online in July 2006 with corrections to the Cover Date.     

In vitro Selection for Tolerance to Fusarium in F1 Microspore Populations of Wheat

Microspore populations of eight Fhybrids of winter wheat (Triticum aestivum L.) whose parents had different levels of resistance to Fusarium were screened in vitro, using phytotoxins of Fusarium as biochemical probe. Two selection methods were compared for the in vitro selection: either embryoids and calli were first initiated from anthers in toxin-free medium and then grown on medium with 0.3—0.9 %Fusarium toxin; or anthers were immediately cultured in modified liquid potato-2 medium in the presence of 1.5, 2.0 and 2.5ml Fusarium toxin per liter culture medium, a concentration which reduced the number of calli and embryoids to about 10 % compared to the toxin-free controls. Microspores from donor hybrids which were produced from very susceptible cultivars were killed by lower toxin concentrations than micro-spores from hybrids of less susceptible parents. From surviving calli and embryoids, originally initiated from 242,000 anthers in both procedures a total of 375 green lines could be regenerated. The results indicate that it is possible to enrich the fraction of regenerating microspores by those which contain the gene complex responsible for reduced susceptibility to Fusarium by the use of a pathotoxin.     

Comparison of Spring Barley Lines Produced by Single Seed Descent, Pedigree Inbreeding and Doubled Haploidy

Samples of inbred lines of Hordeum vulgare produced by pedigree inbreeding, doubled haploidy and single seed descent have been compared for a range of agronomic characters. Differences between the means of the in bred s produced by single seed descent and pedigree breeding were due to differing levels of inbreeding. The present study confirmed chat recombinant inbred families which out-perform the higher scoring parent may be produced by either method. Caution should, however, be exercised when simply inherited characters controlled by major genes are fixed in early generations. Differences between the doubled haploid and single seed descent samples for the character thousand grain weight were attributable to coupling linkages involving epistatic genes. The production and evaluation of random inbred lines should be considered as an alternative to pedigree methods of barley breeding.     

Comparison of doubled haploid lines and F2 bulks for the improvement of barley in the dry areas of North Syria

Drought stress is the main factor limiting barley yields in West Asia and North Africa. This study compares the utility of doubled haploid lines (DHLs) and conventional F₂ plant-derived bulks (F₂Bs) in improving barley in stress environments. Double crosses were made, DHLs were developed by anther culture from double-cross F₁ plants, and F₂Bs were produced by bulking the offspring of F₂ plants. Field tests were conducted in three drought-stressed environments. No major differences were observed in the mean performance of DHLs and F₂Bs. For most traits, both the genotypic and the genotype × location interaction variances were higher in the DHL group, whereas heritabilities were similar. Higher gains from selection were predicted for the DHL group. Regression analysis of yield stability indicated a lower predictability of the DHL performance. The haploid technique can improve breeding populations from which varieties with stable yields can be developed. The costs involved are determined by the DHL production rate, which needs to be improved in many developing countries.     

Plant Regeneration Through Anther Culture of Three Wild Species of Hordeum (H. murinum, H. marinum and H. bulbosum)

Plant regeneration was achieved through anther culture of three wild species of Hordeum (H. murinum, H. marinum and H, bulbosum). Calli or embryoids were formed from microspores in anthers cultured on a medium containing 6-benzylammopurine (BAP) and ficoll. These calli or embryoids regenerated green or albino shoots and roots after transfer to regeneration media. Green plantlets which developed on regeneration media were transferred to soil where they showed further growth.     

Triploid and Hexaploid Regenerants from Hexaploid Timothy (Phleum pratense L.) via Anther Culture

Pollen embryos were obtained from eight genotypes and viable green plants were regenerated from four genotypes in an anther-culture experiment with 165 genotypes of Phleum pratense L. Formation of proembryos inside the cultured anthers during the first 10—12 days was significantly influenced by genotypes and by the type of nutrient media. Primary embryos developed into multiple secondary embryos before regeneration of plants. Among a total of 62 plants regenerated, only 13 were albinos. Of the green regenerants, 11 were triploid while 35 were hexaploid when DNA-content was measured by flow cytometry. Eight plants with a triploid DNA-content did possess the triploid chromosome number of 21. Triploid and hexaploid regenerants from two different parents showed simplified isozyme (GPI and PGD) banding patterns relative to that of their parents.     

In vitro androgenesis of wheat: from fundamentals to practical application

Wheat anther cultures have a history of almost 30 years and are nowemployed efficiently in many countries of the world for the developmentof doubled haploid lines for breeding. The present paper discusses keyquestions of the elaboration and perfection of the method: cytologicalaspects of in vitro androgenesis, the conditions required for theembryogenic development of microspores and the applicability of anthercultures in the Martonvásár wheat breeding programme.     

Potential use of doubled haploid lines for the screening of resistance to yellow rust (Puccinia striiformis) in hexaploid wheat

Doubled haploid lines derived from anther culture of two Iranian spring wheat genotypes‘Ghods’susceptible and‘9106’resistant to yellow rust in Iranian field conditions, and their F₁ hybrids were used in this study. Seedlings of 36 doubled haploid lines, selected out of 96 according to their agronomic traits and the two parental genotypes were inoculated with eight races of yellow rust. The parental genotypes (‘Ghods’and‘9106’) were segregating for some of the races but their doubled haploid lines were either resistant or susceptible to them.‘Ghods’was susceptible to three of the races studied but three doubled haploid lines derived from it were resistant to them. Five selected doubled haploids from the‘9106’genotype and six from F₁ hybrid plants were resistant to all eight races tested. After further investigations in Iranian field conditions it was found that some of these lines can be used as donor genotypes for resistance to yellow rust in wheat breeding programmes. Use of these genotypes should be possible if the French yellow rust races used for selection also represent the dominant races in Iran. It can be concluded that anther culture provides an efficient method for fixing genes of resistance to yellow rust and desirable doubled haploids from F₁ plants can be derived.     

Associations between three ml-o powdery mildew resistance genes and agronomic traits in barley

Summary A population of 198 chromosome-doubled haploid lines of spring barley was scored for segregation in locus ml-o (powdery mildew reaction) on chromosome 4 and in the linked loci s (rachilla hair length) and ddt (reaction to the insecticide DDT) on chromosome 7. They were also tested in a disease-free field trial for the agronomic traits: grain yield, thousand grain weight, lodging, and necrotic leaf spotting. The three mutagen-induced resistance genes ml-o5, ml-o6 (from Carlsberg II) and ml-10 (from Foma) showed no detectable differences with respect to effects on agronomic traits. They all conferred a four per cent reduction in grain yield caused mainly by lower thousand grain weight, and an increase in necrotic leaf spotting. The two original mutants of Carlsberg II had additional mutant genes affecting agronomic traits. Lines with gene S (long hair) had on average a three per cent higher thousand grain weight than those with s. The alleles in locus ddt showed no association with the agronomic traits. It is concluded i) that the associations between the three ml-o alleles and agronomic traits are caused by pleiotropy, ii) that ml-o resistant, high-yielding lines may be selected, and iii) that the association between gene s and thousand grain weight may be due to genetic linkage.Abbreviations DH-lines chromosome-doubled haploid lines