Anther Culture in Rye: Improved Plant Regeneration Using Modified MS-Media

The use of androgenetic doubled haploids in rye breeding is still limited by low regeneration rates. In this study we tested the influence of two modified MS-media on the number of reacting anthers and on plant regeneration. Picloram and 2,4-D were used as auxin components in the induction media. The highest induction rates (reacting anthers per 100 cultivated anthers) and the highest regeneration rates (number of green plants per 100 cultivated anthers) were reached on a modified MS-medium containing Picloram as auxin source, regardless of the donor plant material. The comparison of the results obtained with the single cross ‘SC 35’ which contains genes of Secale vavilovii and the true Secale cereale donor forms clearly show the genotypic influence of donor plant material on induction and regeneration ability.     

The effect of colchicine on triticale anther-derived plants: Microspore pre-treatment and haploid-plant treatment using a hydroponic recovery system

In cereals, chromosome doubling of microspore-derived haploid plants is a critical step in producing doubled haploid plants. This investigation was undertaken to study the effect of incorporation of colchicine in the induction medium for anther culture, and the effect of colchicine on anther culture-derived plants of triticale grown under controlled greenhouse conditions. In the latter case, chromosome doubling of adult sterile plants derived from anther culture of fourteen triticale populations was attempted, where androgenetic plants with non-dehiscent anthers were cloned and subjected to the colchicine treatment, and then grown with the aid of hydroponics. The hydroponic system provided optimal conditions for recovery of the affected haploids from the toxic effects of colchicine treatment and all colchicine-treated plants survived. A topcross-F₁ (TC₁F₁) population with timopheevii cytoplasm produced the highest percentage of plants with seed-set either due to chromosome doubling by colchicine (98%) or spontaneous doubling of chromosome number (15%). Colchicine-treated anthers performed inferior than control in both induction and regeneration phases. One of the key observation of this study was the reversal from reproductive stage back to the vegetative stage which in turn enabled further cloning of haploid plants under hydroponic conditions once they were identified as sterile. The one hundred percent survival rate of in vitro-derived plants, 100% survival rate of colchicine treated haploid plants and the high chromosome doubling success rate (X = 82.3) observed in this study imply that a temperature-controlled greenhouse with an hydroponic system provides an efficient environment for inducing chromosome doubling of haploid plants in cereals. This revised version was published online in August 2006 with corrections to the Cover Date.     

Die Nutzung der Antherenkulturmethode im Zuchtprozeß von Winterweizen

Anther culture in the breeding process of winter wheat. I. Ability of 1B—-1R wheat-rye translocation forms for androgenesis 45 winter wheat varieties or F₁ hybrids, F₂ populations and lines with 1B—1R wheat-rye translocation were tested for their anther culture ability. A total of 48058 anthers was cultured on Potato-2 medium. When averaged over all genotypes and the two experimental years frequencies of embryoid formation of 5.4—6.8 per 100 anthers were observed. Plant regeneration efficiency from embryoids ranged from 5.3—9.1 % or a mean of 4—5 green plants per 1000 anthers plated. The results confirmed the preferential regeneration frequency of gametes with the 1BL—1RS chromosome compared to the gametes with the 1BL—IBS chromosome. Multiple peroxidase were used as marker. The effect of cold pretreatment or of media on the androgenetic response and productivity was not important. On the contrary the variability between the anther response from single ears of the same genotype was noticeable. Examples are presented for the transferability of the androgenetic ability to breeding material. Most green plants obtained were haploid or spontaneous doubled haploid. By cloning it was guaranteed, that progenies were obtained from most of the haploids after colchicine treatment.     

Evidence for genetic control and media effect on haploid regeneration in the anther culture of hexaploid wheat (Triticum aestivum L.)

Crosses were made between seven hexaploid wheat genotypes. Twenty-one F₁ hybrids and their parents were grown in a greenhouse with 16 h day/8 h night at 25°C and 15°C, respectively. The experiment was a complete randomized block design with three replications. Each replication consisted of one pot with three plants. Anther culture was performed in two different induction media (CHB and W14) and androgenetic traits were studied. Statistical analysis was carried out separately for each induction medium. Genetic variation was highly significant for androgenetic traits and the best parent (IBPT 19) produced 68 embryos and 9.3 green plants per 100 anthers in CHB medium. Genetic components were affected by induction media and some components were significant in one medium and non significant in the other. General combining ability (GCA) was significant for all androgenetic traits, except for albino plant regeneration in both media and total plant regeneration in CHB medium, whereas specific combining ability (SCA) was not significant for the traits studied. Narrow sense heritability was high for embryo induction frequency and green and total plant regeneration. All our results indicate that androgenetic parameters can be improved in hexaploid wheat by genetic means.     

Anther callus induction and green plant regeneration at high frequencies from an interspecific rice hybrid Oryza sativa Linn. x O. rufipogon Griff

Summary Callus induction and green plant regeneration at high frequencies from an interspefic hybrid, Oryza sativa L. x O. rufipogon Griff. has been achieved by simply coordinating the growth regulators in the induction medium. The study was conducted with two different basal media (Potato-2 and N₆) and seven different combinations of growth regulators 2,4-D, NAA and kinetin. Synergistic effects of the two auxins in enhanced anther response to callus induction and subsequent green plant regeneration were observed in both media. The highest frequency of callus induction was obtained on Potato-2 medium supplemented with 1 mg/12,4-D, 2 mg/l NAA and 1 mg/l kinetin. The same combination of growth regulators which yielded higher frequencies of callus induction also induced higher mean number of calli per anther. Although the calli formed on N₆ medium showed high regenerability, there was a concomitant increase in the number of albinos among the regenerants. The auxins in the induction media had considerable influence on the regeneration capacity of the calli. The regeneration frequencies were higher from calli formed in the presence of both auxins in the induction media. The levels of growth regulator combinations seem to influence the green plant regeneration especially for calli induced on Potato-2 medium. Among the pollen grain derived plants the majority were either haploids or double haploids and very few were chromosomal variants.     

Characterization of donor genome contents of backcross progenies detected by SSR markers in rice

Backcrossing is a trait introgression method of renewed importance in crops. The evolution of introgressed or substituted segments through backcross generations has been analyzed theoretically using simulations. In this study, the content of donor genomes, including donor segment number (DSN), donor segment length (DSL), and donor genome size (DGS), were directly analyzed in six crosses over three successive backcrosses using a set of single sequence repeat (SSR) markers covering the entire rice genome. The results of this analysis demonstrated that the average DSN in each genome was 8.39 in BC₂F₁, 4.13 in BC₃F₁, and 2.41 in BC₄F₁, decreasing nearly by half with each backcrossing. The average DSL was 33.43 cM (centiMorgans) in BC₂F₁, 29.04 cM in BC₃F₁, and 25.07 cM in BC₄F₁, display a progressive decrease slightly greater than 10% in each additional backcross generation. Meanwhile the average DGS was 280.51 cM in BC₂F₁, 119.97 cM in BC₃F₁, and 60.53 cM in BC₄F₁, decreasing 57.2% from BC₂F₁ to BC₃F₁ and 50.4% from BC₃F₁ to BC₄F₁. This revealed that the reduction in DGS was approximately 50% with each backcrossing. These results provide a guide for introgression or substitution of target chromosome segments from donors into recipients in backcross programs. Zhang-Ying Xi and Feng-Hua He contributed equally to this work.     

Resistance to the cabbage root fly, Delia radicum, within Brassica fruticulosa

Two transgenic Bt rice lines, KMD1 and KMD2, both containing a synthetic cry1Ab gene from Bt, were crossed with conventional rice varieties. The inheritance of resistance to SSB of KMD1 and KMD2was investigated through LSB and field examination of their progenies, e.g. F₁, BC₁ and F₂ populations. In LSBs, 100.0% of newly hatched SSB larvae died on the second day after feeding on leaf tissues of F₁ and GUS positive BC₁ plants, of which the area of leaf tissues consumed by SSB is also similar to that of transgenic parents. These results imply that the resistance of Bt rice to SSB is dominantly controlled and could be easily exploited in hybrid rice production. Field evaluation showed that segregation ratios for SSB resistance to susceptibility in BC₁ populations fit the ratio of 1:1, which was also confirmed by LSBs. However, in F₂ populations, the ratio was significantly smaller than 3:1 for resistant to susceptible plants in all 6 indica × japonica (KMD1 and KMD2) crosses, though it fitted 3:1 in all 4 japonica × japonica crosses. The results implied that the resistance of Bt rice to SSB was controlled by a dominant gene which was present in a homozygous condition in both KMD1 and KMD2, but the inheritance could be affected by other factors. Assays for Cry1Ab protein showed that, in most crosses, the content of Cry1Ab is significantly higher in leaves of GUS positive F₁, BC₁ and F₂ plants than that in transgenic Bt parent plants, which accounts for the high resistance observed in these plants to SSB. This revised version was published online in July 2006 with corrections to the Cover Date.     

Interspecific hybrids between helianthus PetiolarisNutt. and H. Annuus L.: Effect of backcrossing on meiosis

Summary Interspecific substitutions of the nucleus of Helianthus annuus (2n=34) into the cytoplasm of H. petiolaris (2n=34) were obtained by successive backcrossing using cultivated sunflower, H. annuus, as the recurrent pollen parent.Meiosis in the F₁ was characterized by multivalents, suggesting that 10 of the 34 chromosomes were heterozygous for chromosomal interchanges. An additional pair of chromosomes also contained a paracentric inversion. Continued backcrossing resulted in rapid elimination of the meiotic aberrations evident in the F₁. In the BC₁, 1 of 11 plants had normal meiosis and by the BC₂, only 13 of 54 plants had meiotic aberrations similar to those of the F₁. However, trisomic progeny (2n=35) were found in 3 of the 11 BC₁ plants and 20 of the 54 BC₂ plants. No meiotic aberrations were observed in BC₃ or BC₄ plants. Plants with indehiscent anthers, and considered to be male sterile (M.S.), first occurred in the BC₁ and, by the BC₂, 51 of 54 plants were M.S. All 19 BC₃ and 16 BC₄ plants were M.S. Preliminary investigations suggest that the pollen from such plants is sterile and that the sterility is cytoplasmic rather than genetic.Disc-flower measurements were a useful technique for selecting samples at the correct stage of microsporogenesis, but could not be used to distinguish between successive backcrosses.     

Rhizobium specificity of Trifolium ambiguum M. Bieb × T. repens L. hybrid clovers

Growth of full-sib families of an F₁ interspecific hybrid between Trifolium ambiguum M.Bieb and T. repens L, and two generations of backcross hybrids (BC₁F₂ and BC₂F₁) with T. repens as the recurrent parent, were compared to their parental species. Plants were grown in a N-free medium and inoculated with Rhizobium leguminosarum biovar trifolii rhizobia effective on T. ambiguum or T. repens. Hybridisation produced progeny that nodulated with rhizobia from either T. ambiguum or T. repens, but plant growth varied. Mean weights of T. repens and hybrids, particularly F₁ and BC₁F₂, were higher when inoculated with a mixture of strains isolated from field grown T. repens than with the New Zealand inoculant strain for T. repens. When inoculated with the mix of rhizobia from T. repens, mean weights were 242, 189, 132, 125, and 100 mg/plant for T. repens, BC₂F₁, BC₁F₂, F₁ and T. ambiguum, respectively. However, although the mean weight of BC₂F₁ lines was significantly less than T. repens, there was considerable variation in individual full-sib families indicating the potential to select within BC₂F₁ hybrids for high plant growth/symbiotic nitrogen fixation. The weight of T. ambiguumplants inoculated with the New Zealand inoculant strain for hexaploid T. ambiguum was similar to T. repens inoculated with the mix of rhizobia from T. repens (253 and 242 mg/plant, respectively). Mean fresh weights of F₁ hybrid plants were similar when inoculated with rhizobia for T. ambiguum or T. repens (125 and 130 mg/plant, respectively). However, weight of T. repens, BC₁F₂ and BC₂F₁ hybrids inoculated with rhizobia for T. ambiguum were all less than 90 mg/plant. There was a significant relationship between plant fresh weight and ethylene production. The results indicate that measuring weights of inoculated plants growing in N-free media is a rapid initial method of screening a range of plant germplasm for plant growth/symbiotic nitrogen fixation rates. This revised version was published online in August 2006 with corrections to the Cover Date.     

Development of cytoplasmic-genic male sterility in safflower

An interspecific cross was made between Carthamaus oxyacantha and the cultivated species C. tinctorius to develop a cytoplasmic‐genic male sterility (CMS) system in safflower. C. oxyacantha was the donor of sterile cytoplasm. The 3: 1 segregation pattern observed in BC₁F₂ suggested single gene control with dominance of male‐fertility over male‐sterility. The information obtained from crossing male sterile X male fertile plants in BC₁F₃ and BC₁F₄ generations showed statistically significant single gene (1: 1) segregation for male sterility vs. male fertility. The results demonstrated that C. tinctorius possesses a nuclear fertility restorer gene and that a single dominant allele restored fertility (Rf) in progeny carrying CMS cytoplasm of C. oxyacantha. Male sterility occurred with the homozygous recessive condition (rfrf) in a sterile C. oxyacantha cytoplasm background and not in the normal cytoplasm of C. tinctorius. The genetic background of different restorer lines of C. tinctorius having normal cytoplasm did not effect fertility restoration. The absence of male sterile plants in C. tinctorius populations ruled out the possibility of genetic male sterility. Normal meiosis in F₁ and BC₁F₂ ruled out a cytogenetic basis for the occurrence of male sterility.     

Construction and characterization of 3-S Lines, an alternative population for mapping studies in rice (Oryza sativa L.)

Most traits of agronomic importance in rice are quantitative in nature and are controlled by polygenes, called quantitative trait loci (QTL). Understanding the nature and effect of QTLs are important for rice breeding to achieve higher yield and stability. Single segment substitution lines (SSSLs or 3-S Lines) were developed through simple sequence repeats (SSR) marker-facilitated backcrossing methods for Hua-Jing-Xian 74 (HJX74) with the donor segment from six elite germplasm and was characterized. Complete genome survey was carried out with 258 polymorphic SSR markers. Polymorphism of the donors with the recurrent parent varied between 32.98 and 60.73% with an average of 47.81%. Japonica donors were more polymorphic than indica donors. Number of substitution segments per plant decreased with the advancement of backcross generations. In BC₂F_1, BC₃F₁, BC₃F₂ and BC₃F₃ the average number of substitution segment per plant were 12.5, 5.98, 1.69 and 1.46, respectively. Average size of substitution segments also decreased with the number of times plants were backcrossed and selfed. In BC₂F₁, BC₃F₁, BC₃F₂ and BC₃F_3, average size of the segments was 25.43, 22.38, 20.78 and 18.15 cM, respectively. The rate of reduction of segment size was more in backcross (11.99%) than selfing (7.15%) generations. Percent recovery of recurrent parent genome in BC₂F₁, BC₃F₁, BC₃F₂ and BC₃F₃ was 82.24, 92.55, 98.04 and 98.52%, respectively. A total of 111 SSSLs comprising of 43 unique types were developed in BC₃F₂ and BC₃F₃. The estimated length of the segments in SSSLs ranged from 2.00 to 64.80 cM with an average of 21.75 cM, and 6.05 to 48.90 cM with an average of 20.95 cM in BC₃F₂ and BC₃F₃, respectively. Total length of all substitution segments was 2367.5 cM that covered 704.50 cM (39.25%) of the entire rice genome. Effective development and successful utilization of 3-S Lines for analysis of QTLs and mapping of genes established the suitability of the SSR marker facilitated backcross breeding approach for 3-S Lines development and its utilization.     

Cytogenetic study of Helianthus laevigatus and its F1 and BC1F1 hybrids with cultivated sunflower, Helianthus annuus

Although the wild sunflower species Helianthus laevigatus has not been extensively studied it may be considered for sunflower breeding as a potential source of desirable genes for Sclerotinia stalk rot resistance and high contents of proteins and linoleic acid in the seed. A set of six H. laevigatus populations was crossed to cultivated sun~ower lines and produced nine F₁ (2-14 plants) and 66 BC₁F₁ hybrid combinations (1-13 plants). Male sterility occurred in F₁ and BC₁F₁ hybrid combinations and pollen viability was lower in the progenies than in the parents (51.6-77.2%in F₁ and in F₁ and 4.8-34.0% in BC₁F₁). Meiosis was normal in the H. laevigatus populations It was found that this tetraploid species also occurred in a hexaploid form Numerous irregularities were observed in the meiosis of the F₁ interspecific hybrids During diakinesis, quadrivalents and hexavalents were recorded in addition to bivalents Dislocated chromosomes and chromosome bridges were present in the other phases The chromosome number in F₁ was 68 (tetraploid). Irregularities in chromosome pairing were observed in the interspecific hybrids at BC₁F₁. There were many univalents, and trivalents quadrivalents and hexavalents were also present The chromosome number in the BC₁F₁ generation ranged from 34 to 60. The occurrence of meiotic irregularities in the F₁ and BC₁F₁ interspecific hybrids indicates that H. laevigatus and the cultivated sunflower differ in genome constitution.     

Examination and genetic analysis of a yellow-green leaf mutant a269 of sweetpotato

The genetic behaviour of a yellow-green leaf sweetpotato mutant, a269, was analysed to prepare for genetic mapping and cloning of this mutant and to elucidate the regulation of its molecular mechanisms. The phenotypic stability of the mutant was observed under natural conditions, and photosynthetic pigments in mature leaves were measured. GN1323 and a269 were crossed and backcrossed. Genetic behaviour was assessed using phenotypic expression and segregation in F₁, BC₁ and BC₂. The a269 leaves were yellow-green at all stages after germination. The thylakoid lamellae of a269 were disorganized, had incomplete grana and were fewer compared with GN1323. The direct and reciprocal F₁ populations had normal leaf colour, whereas the BC₂ populations had yellow-green leaves. Segregation of plants with normal- and yellow-green leaf colour in the four BC₁ populations satisfied the backcross segregation ratio of 11 : 1 for auto-allohexaploids. The a269 mutant is stable, and its trait is controlled by one recessive nuclear locus.     

Large-scale production of wheat and triticale double haploids through the use of a single-anther culture method

A total of 257 parental wheat and 38 triticale lines were used for anther culture. On average, 2.1 green wheat haploids were obtained per spike. This response occurred irrespective of the origin of the material (Germany, France, Sweden or UK) and 5 years of testing. Triticale responded with 5.3 green haploids per spike. Using the criterion that one parental line should give at least one green haploid per spike in the screening experiment, green haploids were produced from 88 out of 91 F₁ wheat breeding combinations and from each of 21 F₁ and F₂ triticale breeding combinations. An average of 4.7 green plants were obtained per spike from the wheat production programme, while the triticale programme gave an average of 6.2 green plants per spike. A single medium supplemented with different hormones for anthers and embryos was used for culture of both crops.     

Parent genome contribution and its relationship with grain yield in backcross-derived lines of soybean (Glycine max)

Introgression of yellow mosaic disease (YMD) resistance and effect of recurrent parent genome (RPG) on grain yield was studied in 84 soybean genotypes from four populations namely, F_2:7, BC₁F₆, BC₂F₅ and BC₃F₄ derived from cross JS335 x SL525. It was observed that in F_2:7, BC₁F₆, BC₂F₅ and BC₃F₄ derived lines, RPG contribution was 42.5%, 54.9%, 66.4% and 77.6%, respectively, which is significantly less than expected values. Linkage drag from donor parent with YMD resistance gene may be a possible reason for such deviations. Average grain yield per plant in F_2:7, BC₁F₆, BC₂F₅ and BC₃F₄ generations was observed as 13.0, 14.3, 14.9 and 16.1 g, respectively. It was observed that genotypes with more than 80% RPG observed to have both YMD resistance and good yield potential. Graphical genotyping (GGT) analysis revealed that maximum RPG was recovered in chromosomes 8 and 10 and maximum introgression occurred in chromosomes 6 and 19. Our results demonstrated that RPG was positively associated with yield as evident from yield increase with increase in RPG.     

Intergeneric crosses for the transfer of resistance to the beet cyst nematode from Raphanus sativus to Brassica napus

Summary The possibilities to transfer important traits and in particular resistance to the beet cyst nematode (Heterodera schachtii, abbrev. BCN) from Raphanus sativus to Brassica napus were investigated. For these studies B. napus, R. sativus, the bridging hybrid ×Brassicoraphanus (Raparadish) as well as offspring of the cross ×Brassicoraphanus (Raparadish) ×B. napus were used. Reciprocal crosses between B. napus and R. sativus were unsuccessful, also with the use of embryo rescue. Crosses between ×Brassicoraphanus as female parent and B. napus resulted in a large number of F₁ hybrids, whereas the reciprocal cross yielded mainly matromorphic plants. BC₁, BC₂ and BC₃ plants were obtained from backcrosses with B. napus, which was used as the male parent. F₁ hybrids and BC plants showed a large variation for morphology and male and female fertility. Cuttings of some F₁ and BC₁ plants, obtained from crosses involving resistant plants of ×Brassicoraphanus, were found to possess a level of resistance similar to that of the resistant parent. These results and indications for meiotic pairing between chromosomes of genome R with those of the genomes A and/or C suggest that introgression of the BCN-resistance of Raphanus into B. napus may be achieved.     

Production of intergeneric hybrids between Brassica juncea and Diplotaxis virgata through ovary culture, and the cytology and crossability of their progenies

The cytogenetic study was investigated in the intergeneric F₁ hybrid, F₂and backcross progenies (BC₁). The plants used were Brassica juncea(2n=36) and Diplotaxis virgata(2n=18). Three intergeneric F₁ hybrids between two species were produced through ovary culture. They showed 36 chromosomes. It might consist one genome of B. juncea and two genomes of D. virgata. The morphology of the leaves resembled that of B. juncea. The color of the petals was yellow that was like in D. virgata. The size of the petal was similar to that of B. juncea. The mean pollen fertility was15.3% and the chromosome associations in the first meiotic division were(0–1)_IV+(0–2)_III+(8–12)_II+(12–20)_I. Many F₂ and BC₁seeds were harvested after open pollination and backcross of the F₁ hybrids withB. juncea, respectively. The F₂seedlings showed different chromosome constitutions and the range was from 28 to54 chromosomes. Most seedlings had 38chromosomes followed by 36, 40 and 54. The BC₁ seedlings also showed different chromosome constitutions and the range was from 29 to 62. Most seedlings had both 40and 54 chromosomes followed by 36, 46 and52. In the first meiotic division of F₂ and BC₁ plants, a high frequency of bivalent associations was observed in all the various kinds of somatic chromosomes. Many F₃ and BC₂ seeds were obtained by self-pollination and open pollination of both F₂ and BC₁ plants, and by backcrossing both F₂ and BC₁plants with B. juncea, respectively,especially, three type progeny with 36, 40or 54 chromosomes. The somatic chromosomes of the F₃ and BC₂ plants were further investigated. The bridge plants between B. juncea and D. virgata with 36 chromosomes may be utilized for breeding of other Brassica crops as well as B. juncea. This revised version was published online in July 2006 with corrections to the Cover Date.     

Inheritance of black leaf mold resistance in tomato

Summary Inheritance of black leaf mold (BLM) (caused by Pseudocercospora fuligena) resistance was studied in four crosses involving two resistant Lycopersicon accessions (PI134417, L. hirsutum and PI254655, L. esculentum) and four susceptible Asian Vegetable Research and Development Center tomato lines (CLN657BC₁F₂-267-0-3-12-7, CL143-0-10-3-0-1-10, CLN698BC₁F₂-358-4-13 and CL5915-93D₄-1-0-3). For each cross, six generations, i.e. P₁, P₂, F₁, F₂, BC₁F₁ and BC₁F₂ were evaluated following inoculations with isolate Pf-2 of P. fuligena. Chi-square analyses of the data based on the ratio of resistant to susceptible plants in the F₂ in three of four crosses gave a good fit to a segregation ratio of 1 R : 15 S, and BC₁F₂ data in three of four crosses gave an acceptable fit to the segregation ratio of 1 R : 63 S. The results indicate that resistance to BLM may be conditioned by two recessive genes acting epistatically in both PI134417 and PI254655.     

Genetics of colour traits in common vetch (Vicia sativa L.)

Five parents of common vetch (Vicia sativa L.) having orange/beige cotyledon colour, brown/white testa colour, purple/green seedling colour and purple/white flower colour were crossed as a full diallele set. The inheritance patterns of cotyledon, testa or seed coat colour, flower and seedling colour, were studied by analyzing their F₁, F₂, BC₁ and BC₂ generations. The segregation pattern in F₂, BC₁ and BC₂, showed that cotyledon colour was governed by a single gene with incomplete dominance and it is proposed that cotyledon colour is controlled by two allelic genes, which have been designated Ct₁ and Ct₂. Testa colour was governed by a single gene with the brown allele dominant and the recessive allele white. This gene has been given the symbol H. Two complementary genes governed both flower and seedling colours. These flower and seedling colour genes are pleiotropic and the two genes have been given the symbols S and F.     

Die Nutzung der Antherenkulturmethode im Zuchtprozeß von Winterweizen

The use of anther culture in breeding winter wheat. II. Production of new doubled haploid lines of winter wheat with 1AL—1RS-translocation A total of 4472 anhers was cultured from 8 F¹ populations of winter wheat with the 1AL—1RS wheat-rye translocation cultivar ‘Amigo’ as one of the crossing arents. When averaged over all populations a frequency of embruoid formation of 10% and of regeneration efficiency of green plants of 1% were observed. In addition to the 45 green regenerated plants, 93 albinos were obtained. 44% of the green plantlets had 21 chromosome in root tips and 29% were spontaneous diploids. Multiple peroxidases were used a biochemical markers in the subsequent characterization of the homogeneous breeding material. The electrophoretic patterns showed that 16 doubled haploids without rye chromosome segments were produced. In addition the features of dh₀- plants showed, that several of the new 1AL—1RS-translocation lines were awnless.