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.     

Ploidy Manipulation and Introgression of Resistance to Alternaria Helianthi from Wild Hexaploid Helianthus Species to Cultivated Sunflower (H. annuus L.) Aided by Anther Culture

The present investigation discusses the scope for transferring of resistance to leaf spot disease incited by Alternaria helianthi from two hexaploid wild species (H. tuberosus and H. resinosus) to diploid cultivated sunflower. Interspecific hybrids produced between sunflower and these two hexaploid species were partially fertile with tetraploid chromosome status. Backcrosses of these interspecific hybrids with cultivated sunflower resulted in the formation of sterile triploid plants. To overcome the problem of sterility and facilitate backcrosses with cultivated sunflower, anther culture of the tetraploid interspecific hybrids was carried out to bring down their chromosome number to diploid status. Anthers from both interspecific hybrids were cultured on basal Murashige and Skoog media supplemented with varying concentrations of organics and the growth regulators benzyladenine and naphthaleneacetic acid. Anthers of interspecific hybrids involving H. resinosus responded well and regenerated through an embryogenic route at a frequency of 98.7%. But in interspecific hybrids with H. tuberosus, anthers formed callus and subsequently regenerated shoots through an organogenic pathway. DNA ploidy analysis of anther culture plants of interspecific hybrids derived from H. tuberosus crosses was carried out to identify plants with desired diploid status. In vitro screening of parents, interspecific hybrids and anther culture plantlets against A. helianthi showed resistance in 68.5% of the anther culture plants of interspecific hybrids from H. tuberosus and in 24.3% of the plants derived from interspecific hybrids involving H. resinosus.     

Reproductive behaviour and broomrape resistance in interspecific hybrids of sunflower

Interspecific hybrids and backcross generations between the wild perennial species Helianthus resinosus, Helianthus paucifiorus, Helianthus laevigatus, Helianthus nuttallii ssp. nuttallii T. & G. and Helianthus giganteus, resistant to broomrape (Orobanche cernua) and susceptible inbred lines were obtained to study crossability to cultivated sunflower and the transmission and expression of resistance to this parasitic weed. Conventional crosses with all the species tested were successful except for the crosses with diploid H. giganteus, for which embryo rescue techniques were needed to overcome hybrid incompatibility. Pollen viability and seed set were highest for F₁ hybrids with hexaploid species and lowest for those with the diploid H. giganteus. We evaluated F₁, BC₁F₁, some BC₂F₁ plants and the wild and cultivated parents. The wild species and interspecific hybrids were resistant to broomrape infection except for H. nuttallii, which showed segregation, indicating that the resistance is dominant. The crossability and resistance of F₁, and back-cross generations of species with different ploidy levels indicate that the transfer of broomrape resistance to cultivated sunflower is feasible.     

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.     

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.     

Introgression of Cajanus platycarpus genome into cultivated pigeonpea, C. cajan

Summary Cajanus platycarpus, an incompatible wild species from the tertiary gene pool of pigeonpea (C. cajan (L.) Millspaugh), has many desirable characteristics for the improvement of cultivated varieties. To necessitate such transfers, embryo rescue techniques were used to obtain F₁ hybrids. The F₁ hybrids were treated with colchicine to obtain tetraploid hybrids, that were selfed to obtain F₂, F₃ and F₄ progenies. All of the hybrids and subsequent progenies had an intermediate morphology between the two parents. Backcrossing of the tetraploid hybrids with cultivated pigeonpea was not possible given embryo abortion, with smaller aborted embryos than those obtained in the F₀ parental cross.As a route of introgression, diploid F₁ hybrids were backcrossed with cultivated pigeonpea and BC₁ progeny obtained by in vitro culture of aborting embryos. BC₂ plants were obtained by normal, mature seed germination. Although embryo rescue techniques had to be used to obtain F₁ and BC₁ plants, it was possible to produce BC₂ and subsequent generations through direct mature seed. Every backcross to cultivated pigeonpea increased pollen fertility and the formation of mature seeds.Special project assistant till December, 2003.     

Introgression of resistance to Columbia and Northern root-knot nematodes fromSolanum bulbocastanum into cultivated potato

Summary Resistance toMeliodogyne chitwoodi races 1 (MC1) and 2 (MC2) andM. hapla (MH) derived fromSolanum bulbocastanum was introduced into the cultivated potato gene pool through somatic fusion. The initial F₁ hybrids showed resistance to the three nematodes. Resistance to reproduction on roots by MC1 was accompanied by resistance to tuber damage in F₁ clones. Tuber damage sometimes occurred, however, in hybrids of BC₁ progeny resistant to reproduction on roots when MC2 and MH were the challenging nematodes. Resistance to reproduction was transferred into BC₁ individuals, but a greater proportion of BC₁ progeny was resistant to MC1 than to MC2 or MH. Resistance to MC1 appears to be dominant and discretely inherited. F₁ and BC₁ progeny were pollen sterile, but seed were produced from crosses using cultivated tetraploid pollen sources. Approximately 11 and 33 per cent of pollinations produced berries on F₁ and BC₁ pistillate parents, respectively. Seed yield increased fourfold overall in crosses with F₁ compared to BC₁ individuals.Abbreviations MC1 Meloidogyne chitwoodi race 1 - MC2 Meloidogyne chitwoodi race 2 - MH Meloidogyne hapla - Rf Reproductive factor     

Rapid Incorporation of D-genome Chromosomes into A and/or B Genomes of Hexaploid Triticale

Two series of progenies were developed from hybrids between octoploid (AABBDDRR) and tetraploid triticale ((AB)(AB)RR). One arose from the successive selfing of the F₁s, while the second was established after one backcross of the F₁ hybrids with the respective 8 × triticale parent. Altogether, 250 F₃ and BC₁F₂ lines were developed, of which 112 were karyotyped in the F₄/F₅ or in BC₁F₃/BC₁F₄ generations using C-banding and SDS-PAGE. The 112 lines represented 61 different karyotypes, of which 39 appeared to be stabilized, having pairs of homologous wheat chromosomes only, while 22 karyotypes exhibited 1—3 heterologous pairs. The frequency of karyotypically stabilized lines originating from the series with one backcrossing was much higher (79.5 %) than those derived from the successive selfing of the F₁ hybrids (51.7%). Six lines had the pure hexaploid triticale chromosome constitution. The frequency of disomic substitutions of D genome chromosomes for their homoeologous A and/or B genome chromosomes ranged from one to six per line with an average of 1.7. Except for 3B and 6B all possible D(A/B) substitutions were obtained. Chromosomes ID and 3D substituted for their homoeologues with the highest frequency, while the substitution of chromosome 4D for 4A or for 4B was the least frequent. D(R) substitutions were found in eight lines only. A complete set of 6x triticale lines was established in which chromosome ID was present in all possible combinations, i.e. single 1D(1A/1B/1R) disomic substitutions as well as disomic ID addition.     

Performance Studies in F6 Lines of Autotetraploid Fenugreek

Hybridization followed by pedigree selection was carried out between two autotetraploid strains of fenugreek (Trigonella foenum-graecum L.). Morphological and cytological investigations were carried out in two autotetraploid parents, their F₁ hybrids and F₆ lines derived from F₁ after selfing. Cytomorphological data of F₆ lines were compared with those of F₁, hybrids and of their parents. The hybrid plants were intermediate with respect to different vegetative traits, however, they showed increased seed set as compared to their autotetraploid parents. The mean pod number per plant in F₆ was one and half times as high as one of the high yielding tetraploid parents and approximately twice as high as that of F₁ hybrid. Meiosis in F₆ was more regular than in F, and the parents. Thus, hybridization followed by pedigree selection is helpful not only in regularizing the meiotic behavior, but vigour and seed fertility have also improved in autotetraploid fenugreek.     

Variation for aphid resistance and insecticidal acyl sugar expression among and within Lycopersicon pennellii-derived inbred backcross lines of tomato and their F1 progeny

Five inbred backcross lines (IBL) were selected for higher relative expression of insecticidal acyl sugars (rank average) from an inbred backcross population derived from the cross Lycopersicon esculentum cultivar ‘Peto 84’×Lycopersicon pennellii accession LA716. These five BC₂S₅ IBLs were crossed in a partial diallel design (Method II), and their self and F₁ progeny and three control cultivars were tested at two California field locations in 1996. Counts of potato aphids, Macrosiphum euphorbiae Thomas, on leaves, as a measure of plant infestation, revealed significant general combining ability (GCA) for lower aphid numbers with IBL44 and IBL59; the F₁ hybrid IBL44 × IBL59 had significantly fewer aphids per leaflet than the susceptible cultivar ‘Alta’. GCA for acyl sugars was associated with IBL59 only. Of all the IBL and IBL × IBL F₁ hybrids, only IBL59 produced significant levels of acyl sugars. Significant within IBL59 variation for acyl sugars was observed, but not for aphid resistance. Our results suggest that factors other than acyl sugars contributed to L. pennellii-derived aphid resistance in IBL × IBL F₁ hybrids and IBL that do not produce significant amounts of acyl sugars. IBL59 and IBL44 may be useful for breeding for aphid resistance in cultivated tomato.     

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.     

The transfer of triazine resistance from Brassica napus L. to B. oleracea L. III. First backcross to parental species

Summary Atrazine resistant Brassica napus × B. oleracea F₁ hybrids were backcrossed to both parental species. The backcrosses to B. napus produced seeds in both directions but results were much better when the F₁ hybrid was the pollen parent. Backcrosses to B. oleracea failed completely but BC₁s were rescued by embryo culture both from a tetraploid hybrid (2n = 4x = 37; A₁C₁CC) and sesquidiploid hybrids (2n = 3x = 8; A₁C₁C). Progeny of crosses between the tetraploid hybrid and B. oleracea had between 25 and 28 chromosomes. That of crosses between the sesquidiploid hybrid and B. oleracea had between 21 and 27. A few plants that had chromosome counts outside the expected range may have originated from either diploid parthenogenesis, unreduced gametes or spontaneous chromosome doubling during in vitro culture. Pollen stainability of the BC₁s ranged from 0% to 91.5%. All the BC₁s to B. oleracea were resistant to atrazine.     

Transfer of architectural traits from perennial Helianthus mollis Lam. to sunflower (H. annuus L.) and localisation of introgression

Common sunflower (i.e., Helianthus annuus) is occasionally crossed with other Helianthus species to gain favourable agronomic traits. An interspecific cross between H. annuus and Helianthus mollis was used to examine events that occurred during the five first generations. First generation hybrid plants were verified using GISH, and F₁ hybrid (#17) was back-crossed to common sunflower to obtain BC₁ (#4) and BC₂ (#21). Some BC₂ progeny possessed new traits, i.e., compact (Compact plant architecture is due to short inter-node length, short petiole length and short leaf lamina length) architecture without and with corrugated leaves, which were not observed in the parents. These traits were selected for and fixed after the BC₂-S₃ (#18) to produce a H. mollis introgression line (namely HM374). Although the size of the population was small, we tentatively construct a genetic map using data from the BC₂ generation. Two linkage groups were identified carrying H. mollis specific AFLP markers that were associated with compact plant architecture, and one unlinked marker was shown to explain variation for the corrugated leaves. Introgression line HM374 was used to test the inheritance of compact plant architecture in a cross with an elite high oleic line, OL75HO. Microsatellite (SSR) markers were used to genotype F₁ and F₂ offspring’s. Our data suggested that the line HM374 was still heterozygous for introgression after eight generations. HM374 displays new architecture traits (i.e. compact architecture with or without corrugated leaves) and carries H. mollis fragments anchored on LG1 and LG11 of sunflower public map.     

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.     

Tolerance to low temperatures and tuber soft rot in hybrids between Solanum commersonii and Solanum tuberosum obtained through manipulation of ploidy and endosperm balance number (EBN)

The objectives of this study were to evaluate the tolerance to low temperatures and tuber soft rot in hybrids between Solanum commersonii and Solanum tuberosum. The experimental materials consisted of F₁ triploid, BC₁ pentaploid‐near pentaploid and BC₂ tetraploid–near tetraploid hybrids. The F₁ triploids had a freezing tolerance and acclimatization capacity closest to S. commersonii. This indicated that the endosperm barriers which prevent the introgression of 1EBN S. commersonii into 4EBN S. tuberosum had been overcome. Indeed, the triploids produced 2n eggs, thus giving a compatible maternal to paternal EBN ratio in the hybrid endosperm generated by the 3x(2EBN) × 4x(3EBN) crosses. The tolerance to low temperatures of BC₁ and BC₂ hybrids was lower than that of the F₁. However, a number of genotypes were identified which were able to withstand temperatures down to ‐5°C. Some BC₂ hybrids were also tested for their tolerance to tuber soft rot, and some resistant hybrids were detected. A number of them combined the capacity for cold acclimatization with tolerance to tuber soft rot. These hybrids have an EBN of 4; they are fertile and have been used in backcrosses with 4EBN S. tuberosum.     

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.     

Comparing mating designs to restore seed production of interspecific hybrids between Trifolium repens (white clover) and Trifolium uniflorum

Interspecific hybrids between Trifolium uniflorum and cultivated white clover (Trifolium repens) have highly useful characteristics for temperate pastoral systems derived from both parent species. However, the early hybrids (F₁ and BC₁) also have unacceptably poor seed production for commerce. This study analysed the basis for the poor seed production and investigated breeding strategies for overcoming the problem. The BC₁F₁ generation produced lower‐than‐expected numbers of heads per plant and seeds per floret. Backcrossing of selected hybrids to white clover corrected these deficiencies and created new variation. Seed numbers were also returned to near target levels by recurrent selection within the BC₁ generation. Thus, it was possible to retain a theoretical average of 25% of T. uniflorum genome and still achieve high seed production per plant. The BC₁F₂ and BC₂F₁ generations produced high seed numbers per plant, along with reasonable variation. Both of these second‐generation hybrid forms have high reproductive potential and should be the focus for the selection of the desired combinations of agronomic and seed production traits.     

Introgression from wild Cicer reticulatum to cultivated chickpea for productivity and disease resistance

Interspecific hybridization is known to improve productivity and resistance to diseases in many crops. Therefore, an attempt was made to introgress productivity and disease resistance into chickpea from wild Cicer species. The true F₁ hybrids of cultivated chickpea genotypes ‘L550’ and ‘FGK45’ with C. reticulatum were backcrossed twice to their cultivated female parents to minimize the linkage drag of undesirable wild traits. The pedigree method was followed to advance the segregating populations from straight crosses (without backcross) and BC₁/BC₂ generations to F₅–F₇. The interspecific derivatives recorded up to a 16.9% increase over the check cultivars and a 25.2% increase over the female parent in a preliminary yield evaluation trial. Of the 22 interspecific derivatives thus derived, four desi and two kabuli lines were further evaluated for seed yield in replicated trials at three diverse locations. These lines possess a high degree of resistance to wilt, foot rot and root rot diseases, and recorded a 6.1–17.0% seed yield increase over the best check cultivars.     

Intergeneric hybridization of Diplotaxis erucoides with Brassica napus. I. cytogenetic analysis of F1 and BC1 progeny

Summary Intergeneric hybrids (F₁) Diplotaxis erucoides (DeDe) x Brassica napus (AACC) and the first backcross to B. napus (BC₁) have been obtained through in vitro culture of excised ovaries. The chromosome numbers of F₁ and BC₁ plants proved the occurrence of unreduced gametes. The study of metaphase I chromosome pairing showed that autosyndesis in De genome and allosyndesis between De and A/C genomes might exist. The male fertility of the F₁ plants was low. Some male-sterile plants were found in F₁ and BC₁ progeny. The possibilities of creating addition lines B. napus-D. erucoides and of obtaining a new cytoplasmic male sterility in B. napus are discussed.     

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.