Cytogenetic studies in hexaploid Helianthus species and their F1 hybrids with cultivated sunflower, H. annum

Hexaploid sunflower species provide potential sources of desirable genes in breeding for resistance to pathogens as well as high protein and oleic acid content. The possibility of using them in sunflower breeding was studied by testing the crossability of hexaploid species to cultivated sunflower, by the analysis of meiosis and pollen viability in the F₁ interspecific hybrids. The hybrids were obtained from 16 combinations, with 1–18 F₁ plants per combination. The F₁ hybrids in general exhibited the dominant phenotype of the wild species. Sterility ranged from 0 to 87.5% per F₁ combination, while pollen viability ranged from 43.0 to 70.2%. Meiosis was almost normal in the species analysed but irregular in the F₁ hybrids. Most of the meiocytes showed predominantly bivalents (86.79–90.22%), but univalents (0.00–4.99%) and the multivalents (8.23–10.09%) occurred as well. At anaphase I, chromosome bridges were detected in 6.88–8.48% of meiocytes. Fast chromosomes in meta-phase I and lagging chromosomes in anaphase I and telophase II were observed in many meiocytes.     

Meiosis and pollen grain viability in Helianthus mollis,Helianthus salicifolius,Helianthus maximiliani and their F1 hybrids with cultivated sunflower

Three diploid perennial sunflower species are useful for variety improvement: Helianthus mollis, because of sessile leaves, H. salicifolius, because of a high oil concentration, and H. maximiliani, a potential source of resistance to Sclerotinia sclerotiorum. The crossability of these species to cultivated sunflower was examined.Hybrids were obtained from eight combinations, with 3–15 F₁ plants per combination. The F₁'s exhibited the dominant phenotype of the wild species. Pollen viability varied between 32.1 and 69.9%. Meiosis was irregular in the F₁ hybrids. At diakinesis, bivalents (62.7–97.9% of meiocytes), univalents (0–31.23%), and multivalents (3.84–7.68%) were detected. At anaphase I, chromosome bridges were detected in 6.77 to 11.44% of meiocytes. Fast chromosomes in metaphase I, and lagging chromosomes in anaphase I and telophase II were evidenced in a high percentage of meiocytes.     

Effect of wild Helianthus cytoplasms on agronomic and oil characteristics of cultivated sunflower (Helianthus annuus L.)

Sunflower (Helianthus annuus L.) productions reliance on a single source of cytoplasmic male sterility, PET1, derived from H. petiolaris Nutt., makes the crop genetically vulnerable. Twenty diverse cytoplasmic substitution lines from annual and perennial wild species were compared with the inbred line HA89 over four environments. Pairwise comparisons of alloplasmic male fertile (MF) with euplasmic MF lines suggested that cytoplasm of perennial H. angustifolius increased lodging while perennial cytoplasms of H. mollis, H. grosseserratus, and H. divaricatus reduced head size and consequently reduced yield and will need to be used with caution. Lines having annual species, cytoplasms had no effects on agronomic traits. Comparisons of CMS HA 89 with 10 CMS lines suggested yield‐reducing cytoplasmic effects of perennial H. maximiliani and annual H. annuus PI 413178 and PI 413024. Cytoplasmic effects on oil percentage and fatty acid composition were minimal and should not be of concern for sunflower breeders. In general, most cytoplasms of wild annual Helianthus species accommodate cultivated nuclear genes without drastic adverse interactions, and are potential sources of cytoplasmic diversity for sunflower breeding.     

Diversity among sources of cytoplasmic male sterility in sunflower (Helianthus annuus L.)

Summary Ten cytoplasmic male sterile (CMS) sunflower (Helianthus annuus L.) lines were crossed with nine maintainer or male fertility restorer lines in a diallel crossing scheme. Based on fertility restoration of the F₁ generation, CMS lines were divided into four groups. At least two new sources of CMS, CMS PET2 and CMS GIG1, were found to be potentially useful for commercial production of hybrids. Environment had an influence on fertility restoration of one CMS line, CMS MAX1. Effective restoration of male fertility for CMS RIG1, CMS ANN2, and CMS ANN3 was not found.     

Cytogenetic analysis of F1, F2 and BC1 plants from intergeneric sexual hybridization between Sinapis alba and Brassica oleracea by genomic in situ hybridization

By intergeneric sexual hybridization between Sinapis alba and Brassica oleracea , F₁, F₂ and BC₁ progeny plants were produced. S. alba plants (genome SS, 2n = 24) were pollinated with B. oleracea (genome CC, 2n = 18), and the fertile F₁ plants were pollinated with B. oleracea to obtain BC₁ plants. GISH analysis showed that 10 out of 12 F₁ plants had 12 S. alba chromosomes (one full S chromosome set) and nine B. oleracea chromosomes (one C chromosome sets), representing the expected hybrids. However, two F₁ plants had 12 S chromosomes and 18 C chromosomes (two C chromosome sets), indicating unexpected hybrids. A maximum of three trivalents between C and S chromosomes were identified at metaphase I of semi-fertile F₁ pollen mother cells (PMCs), which indicates homology and chromosome pairing between these two genomes. The C genome had obviously been doubled in two F₂ plants from selfed semi-fertile F₁ plants. BC₁ plants consisted of 18 C chromosomes and different numbers of one, five and six additional S chromosomes, respectively. Monosomic alien addition lines developed in the present study can be used for B. oleracea breeding and Sinapis alba gene mapping.     

Interspecific hybrid of Helianthus annuus × H. simulans: Characterization and utilization in improvement of cultivated sunflower (H. annuus L.)

The first success at interspecific hybridization between cultivated sunflower(H. annuus) and a diploid perennial species, H. simulans is reported. The F₁s from both direct and reciprocal directions exhibited dominance of the wild species phenotype and were pollen sterile. Meiosis was irregular in the F₁ hybrids and both univalents and multivalents were observed. Multiplication of the interspecific hybrids was achieved through in vitro culture of nodal sections on Murashige & Skoog medium supplemented with 0.5 mg l^-1 benzyladenine. Fertility of the interspecific hybrids was improved by subjecting the in vitro proliferating shoots to 0.001% colchicine incorporated in the shoot multiplication medium. The amphiploids serve as fertile bridges and facilitate interspecific gene transfer through conventional breeding. This revised version was published online in July 2006 with corrections to the Cover Date.     

Morphological and cytological study in a new type of cytoplasmic male-sterile line CMS-GIG2 in sunflower (Helianthus annuus)

Cytoplasmic male sterility (CMS) is essential for sunflower hybrid production. CMS-GIG2, a new sunflower CMS type, was further confirmed by crossing with the maintainer and restorer lines for the CMS-PET1, both of which maintain the male sterility of CMS-GIG2. Meiotic division in CMS-GIG2 was observed with 4'6-diamidino-2-phenylindole dihydrochloride staining, indicating that microspore formation was disrupted before the meiotic cytokinesis was completed. Light microscopy observation showed that both middle layer and tapetal cells expand radially rather than degrade over time, followed by failure to form tetrads and normal microspores. This morphological defect leading to male sterility in CMS-GIG2 differs from that observed in the PET1 CMS type. CMS-GIG2 will certainly provide additional genetic diversity for sunflower hybrid breeding programmes.     

Heterosis for yield and other agronomic traits of winter triticale F1 and F2 hybrids

Triticale is generally treated as a self‐pollinating crop and line breeding is practised. Hybrid breeding has been discussed for some time, but there is little information for winter triticale. This study investigated heterosis for eight agronomic traits in F₁ and F₂ hybrids grown together with their parents as drilled plots in three environments. On average, grain yield heterosis was 12.5 dt/ha (a relative 10.5%) compared with the mid‐parent value for F₁ hybrids, and 6.2 dt/ha (5.0%) for F₂ hybrids and withawide range of 4.4–17.1 dt/ha for F₁ hybrids. A positive contribution to the heterosis of yield was made by kernels/spike and 1000‐kernel weight, whereas spikes/m² showed negative heterosis. Hybrid plants in F₁ and F₂ were taller than mid‐parents (8.3 cm and 5.3 cm, respectively), with a tendency to earlier heading. The negative heterosis for falling number in F₁ and F₂ hybrids could be a problem for commercial production of triticale hybrids. By selecting parents for combining ability and the identification of heterotic patterns, grain yield heterosis of 20% appears feasible.     

Meiosis and Pollen Viability in Helianthus tuberosus L. and its Hybrids with Cultivated Sunflower

The hexaploid species of Helianthus tuberosus is a potential source of resistance to several sunflower pathogens. But crossed with cultivated sunflower, it produces F, hybrids which have low values of fertility or even full sterility. Pollen viability and meiotic features were studied in 17 populations of the species H. tuberosus and in F¹ hybrids. Significant differences in pollen viability existed between populations (47.1–98.8%). In the 15 FI hybrids, plant fertility ranged from 0 to 100 per combination, while pollen viability ranged from 12.4 to 57.1. Meiosis was almost normal in the analyzed species, and irregular in the F, hybrids. The highest percentage of meiocytes was with bivalents (85.9), but univalents (0.3) and multivalents (13.8) occurred as well. In metaphase and anaphase, the percentage of meiocytes with fast and lagging chromosomes was high. In anaphase, chromosome bridges were detected in 9.9 of the meiocytes.     

Heterosis performance of yield and fibre quality in F1 and F2 hybrids in upland cotton

Because of the difficulty of producing F₁ hybrid seeds by hand emasculation and pollination, wide use of heterosis in cotton production has been limited in China. The objective of this study was to evaluate the potential of F₂ hybrids for yield and fibre quality. A half diallel involving eight parents and their F₁ and F₂ hybrids was grown in replicated studies at Linqing and Nanjing in 1999 and Nanjing in 2000. Yield and fibre quality was determined for all 64 entries. Fibre quality was also determined for parents and F₁s, but only for Zhongmiansuo 28 (ZMS28), Xiangzamian 2 (XZM2) and Wanmian 13 (WM13) F₂s. These three F₂ hybrids are extensively planted in China and provide experimental controls with which to compare the performance of new hybrids. Average yield heterosis for F₁s and F₂s was 15.9 and 9.2%, respectively. Inbreeding depression for yield varied but some F₂s greatly out‐yielded the best variety. Average F₁ heterosis was 6.7, 6.2 and 2.9%, respectively for number of bolls per unit area, boll weight, and lint percentage. The average F₂ heterosis for the same traits was 4.4, 3.3 and 1.6%, respectively. F₁ heterosis for fibre traits was low. In general, parental average was a good indicator of the yield and fibre quality of F₁ hybrids. These encouraging results suggest there is sufficient heterosis for yield to use F₂s in China.     

Genetic variation and changes with ontogeny of osmotic adjustment in sunflower (Helianthus annuus L.)

Summary Intraspecific variation for osmotic adjustment in sunflower was examined using a collection of 33 genotypes of different origin which were exposed to water stress at the 8-leaf stage. Changes in osmotic adjustment with ontogeny were also evaluated in the pre- and post-anthesis phases using seven genotypes drawn from this collection. Estimates of osmotic adjustment were derived from measurements of leaf relative water content (RWC) and osmotic potential () during a period in which the soil was allowed to dry out gradually. The degree of osmotic adjustment, expressed as the value of RWC for a of –1.7 MPa (RWCe), was derived from the ln RWC/ln relationship. Both monophasic and biphasic ln RWC/ln relationships were found. Irrespective of the form of the relationship, all genotypes at the 8-leaf stage showed some degree of osmotic adjustment. This was also true for the cultivars included in the subset examined in pre- and post-anthesis phases. Significant differences (P=0.05) in RWCe were found between extreme genotypes in all three phases.Significant (P=0.05) linear relationships were found between RWCe measured in the 8-leaf stage and that measured in the pre- and post-anthesis phases, establishing the viability of measurements in the 8-leaf stage as a means of selection for osmotic adjustment in later developmental stages. Genotype rank order was stable (P=0.01) across the three ontogenetic phases examined.Abbreviations ETp potential evapotranspiration - osmotic potential - RWC relative water content - RWCe value of RWC for a of –1.7 MPa     

Yield performance of interspecific F1 hybrids of the greek mountain tea, Sideritis spp. L.

Interspecific F₁ hybrids were produced after artificial intercrossing of six Sideritis species indigenous to Greece, namely: S. athoa, S. euboea, S. syriaca, S. raeseri, S. scardica, S. clandestina and a cultivated clone of S. raeseri. It was found that all species cross with each other, but the level of crossability varied with the cross-combination. In the yield evaluation experiments, the interspecific hybrids with a common female parent were considered as a group. It was observed that yield during the year of establishment was relatively low, but the interspecific hybrids out-yielded the control in both fresh and dry weight. All hybrids produced higher yields in the following years and their productivity started to decline after the fifth year. All hybrids produced higher yields than the control in all six years of the experiment. The highest yield over the years, in fresh and dry weight, was produced by the group Sideritis raeseri cultivated clone (cc) ×Sideritis spp. This yield, however, was not always significantly different from the yield produced by the second in the rank group (S. clandestina×Sideritis spp.). A high correlation (r > 0.99) between fresh and dry weight was observed. The two groups with the highest yield also had the highest essential oil content. In addition, all the interspecific hybrids had a higher essential oil content than the control species. Furthermore, the essential oil content of the selected individual hybrid plants ranged between the midparental value to a value higher than the high-parental species, indicating an additive and dominance gene action. It was also found that the content of the essential oil in the seven most important components (pinene, lemonene, cynene, menthone, copaene, caryophyllene and valerianic ester) varied from hybrid to hybrid. Given that Sideritis reproduces mainly asexually, the direct use of individual interspecific hybrids with higher yield and content in essential oil than the control species is possible in cultivation.     

Cytogenetic Studies of Crosses between Lolium multiflorum Lam. and Festuca arundinacea Schreb. III. The Generations C1, C2 and C3

Meiotic observations of 56-chromosome plants of the C₁, C₂ and C₃ generations of the amphidiploid Lolium multiflorum × Festuca arundinacea showed that most of the chromosomes paired as bivalents. However, uni- and multivalents were frequently observed. No difference in meiotic regularity and stability could be detected between the different generations. The average chromosome number and; the percentage of 56-chromosome plants decreased with increasing generations. The direct use of the 56-chromosome amphidiploids of L. multiflorum and F. arundinacea in a breeding programme appears to be quite difficult, but these plants can be used in a backcross programme with Lolium and Festuca.     

Segregation patterns of AFLP markers in F1 hybrids of a cross between tetraploid and diploid species in the genus Malus

Malus xiaojinensis, one of the most important wild genotypes in the genus Malus, is resistant to a variety of stresses such as Fe deficiency chlorosis, drought and cold. However, lack of knowledge of its genetic background prevents using genetic analysis to study those agronomic traits and corresponding gene functions. Here, as the first step towards construction of the linkage map of M. xiaojinensis, genetic analysis of the F1 triploid hybrids (M. xiaojinensis × M. baccata) was performed with amplified fragment length polymorphism (AFLP) markers. Using 15 EcoRI‐ MseI primer combinations, 1110 AFLPs were identified, with 31.3% of M. xiaojinensis‐, 12.7% of M. baccata‐specific markers, 54.9% of common markers, and 1.2% of non‐parental markers; 93.3% of the AFLP markers exhibit the expected segregation ratio. Thirty‐two M. xiaojinensis‐specific markers and 47 common markers display a 5 : 1 and 11:1 segregation ratios, respectively, suggesting that M. xiaojinensis is an autotetraploid, or at least an isosyndetic allotetraploid.     

Comparison and integration of genetic maps generated from F2 and BC1-type mapping populations in perennial ryegrass

Linkage maps of perennial ryegrass were constructed from F₂ and BC₁‐type populations using, predominantly, restriction fragment length polymorphism data based on heterologous probes used in mapping other grass species. The maps identified seven linkage groups, which covered a total of 515 cM (F₂) and 565 cM (BC₁). They were aligned using 38 loci identified in both populations (common loci) and a possible marker order for all mapped loci in either population was identified in an integrated map. The estimated recombination frequencies and map distances between adjacent common loci were compared between the two data sets and regions of heterogeneity identified. Overall, the common markers identified a map distance of 446 cM in the F₂ population and 327 cM in the BC₁ population, reflecting a higher recombination frequency in the former, although the difference was not evenly spread over the seven linkage groups.     

Relationship of Root and Shoot Characters in Parent, F1 and F2 Populations of Rice

The direct and indirect contributions of root characters — root length, roots/plant, fresh and dry root weight on grain yield/plant were worked out from a 7 × 7 diallel set of rice hybrids. The materials were grown in pots with four replications. Path analysis was done at genotypic level of correlation.
The grain yield/plant showed positive correlation with all the root characters in parent, F₁ and F₂ except with roots/plant in F₁ population. Fresh root weight demonstrated positive direct effect on grain yield/plant in all the three generations. Roots/plant had highly positive direct effect in F₂. Direct effects were negative in respect of root length and dry root weight in F₂ generation.     

The use of AFLP markers for the identification of carrot breeding lines and F1 hybrids

Four inbred lines of carrot (cytoplasmic male‐steriles and corresponding maintainers) and eight of their F₁ hybrids were studied with the amplified fragment length polymorphism (AFLP) technique to examine their genetic relationship and produce markers useful for testing hybrid seed purity. Eighty‐six polymorphic amplicons were identified in bulked DNA samples using eight primer pair combinations. Genetic distance was estimated on the basis of the presence or absence of polymorphic bands. The dendrogram plotted on the basis of the AFLP data closely represented the pedigree relationships of the lines and their hybrids. From one to six amplicons specific for a breeding line were identified. Most of them were also present in the DNA bulks of respective F₁ hybrids. However, screening performed on individual plants of two parental lines and the corresponding hybrid indicated insufficient uniformity of parental lines, limiting the applicability of AFLP markers for testing hybrid seed purity.     

Effect of parental genotypes and colchicine treatment on the androgenic response of wheat F1 hybrids

The effect of the parental genotypes and colchicine treatment on the androgenic response of wheat (Triticum aestivum L.) F1 hybrids was studied. For this, anthers from three F1 hybrids and their parents were cultured on W14 initiation medium and W14 supplemented with 0.03% colchicine. The number of responding anthers, microspore‐derived structures/100 anthers, green plants/embryos cultured, green plants/100 anthers and albino plants/100 anthers were recorded. It was observed that embryo formation and plant regeneration ability were genetically controlled and genotype dependent. In both treatments the variety Kavkaz had a significantly higher percentage of responding anthers, microspore‐derived structures and green plants/100 anthers than the other genotypes. On the other hand, the variety Myconos also demonstrated high microspore‐derived structure production and green plant regeneration when treated with colchicine. The good response observed in these two varieties indicates the importance of colchicine treatment only for certain genotypes. Green plant production capacity of the hybrids was intermediate to that of the parental varieties. As one parent with a high or even an intermediate response to anther culture could lead to the production of sufficient (for breeding purposes) green plants from the F₁ hybrids, it was concluded that screening the inbred lines for the response to anther culture with and without colchicine treatment could contribute to utilization of breeding material with a low response to anther culture via the proper hybrid combinations.     

Anther-culture response in different genotypes and F1 hybrids of pepper (Capsicum annuum L.)

An in vitro anther-culture method has been improved by using young mother plants and by using frequent subcultures, thus increasing the androgenic yield in different Capsicum annuum L. genotypes. An assortment of peppers was used, composed of 15 genotypes (four breeding lines, seven cultivars and four F₁ hybrids). A new system for qualifying the androgenic response was established. For use in practical breeding, a minimum of 5 % of plant regeneration was proposed as the criterion for a fair response. Accordingly, one excellent, one good and eight fair responses were identified among the genotypes investigated. As compared to the standard cultivar. 2 genotypes gave a significantly better response, i.e. ‘Fehérözön’ (75.8%) and ‘Szechuan 90716’ (21.0%). In comparative investigations, F₁ hybrids, produced from crosses between poor/non-responsive and responsive genotypes, showed a fair level of response, even the case of a poor response in donor parent. The ploidy level of the resulting plants was determined by flow-cytometric analysis.     

Evaluation of F2 and F3 plants introgressed with QTLs for clubroot resistance in cabbage developed by using SCAR markers

Clubroot disease caused by Plasmodiophora brassicae is one of the major diseases of Brassica crops, often devastating to the cultivation of cruciferous crops in temperate regions. In a previous study (Moriguchi et al. 1999) identified three major quantitative trait loci (QTLs) for clubroot resistance, each in a separate linkage group, in a population derived from a cross between a clubroot‐susceptible inbred cabbage line, Y2A and a resistant inbred kale line, K269. In this study, the original random amplified polymorphic DNA (RAPD) and restriction fragment length polymorphism (RFLP) markers were converted into sequence‐characterized amplified region (SCAR) markers to facilitate large‐scale marker‐assisted screening of clubroot resistance in cabbage breeding. Of 15 RAPD markers closely linked to the three QTLs, nine SCARs were developed as dominant markers after cloning and sequencing. In addition, two RAPD markers were converted into co‐dominant cleaved amplified polymorphic sequence (CAPS) markers, and one RFLP marker out of three tested was converted to a dominant SCAR marker. The effect of selection for resistance by the improved markers was evaluated in progeny plants in the F2 and F3. A total of 138 F2 plants were genotyped with nine SCARs and 121 well‐distributed makers consisting of 98 RAPD, 19 RFLP, two isozymes, and two morphological markers in order to estimate the level of resistance and the proportion of undesirable alleles from the kale in non‐target areas in each of the F2 populations. An F2 plant, YK118, had kale alleles at QTL1, QTL3 and QTL9. Three F2 plants, namely, YK107, YK25 and YK51 had kale alleles at only QTL1, QTL3 and QTL9, respectively. These F2 plants were selected for their low proportion of alleles derived from kale in non‐target regions. YK118, like the resistant kale parent, expressed very high resistance to three field isolates of Plasmodiophora brassicae, whereas the mean disease index in the F2 and F3 plants carrying only single QTLs was intermediate. The QTLs showed no differential response to the isolates. These plants with improved resistance will be useful as parental inbred lines for F1 hybrids.