Relative efficiency of maize and Imperata cylindrica for haploid induction in Triticum durum following chromosome elimination‐mediated approach of doubled haploid breeding

Intergeneric hybridization in seven diverse durum wheat genotypes was carried out using two composite varieties of Himalayan maize, viz., Bajaura Makka and Early Composite, and a wild grass, Imperata cylindrica, as pollen sources. Observations related to various haploid induction parameters put forth I. cylindrica as significantly better pollen source for haploid induction in durum wheat over maize in terms of pseudoseed formation (46.93%), embryo formation (38.06%), haploid regeneration (40.42%) and haploid formation efficiency (7.44%). The line x tester analysis revealed that both male and female genotypes had significant effects on all haploid induction parameters except haploid formation frequency in later. Among the pollen sources, I. cylindrica emerged as best combiner based on GCA values when compared with the two Himalayan maize composites. Durum wheat genotype, A‐9‐30‐1 was recognized as the best general combiner followed by PDW 314. The present investigation proposed durum wheat × I. cylindrica as a superior technique over maize‐mediated system, and its large‐scale use can open a new horizon in the sphere of durum wheat doubled haploidy breeding programme.     

Influence of winter and spring wheat genetic backgrounds on haploid induction parameters and trait correlations in the wheat × maize system

The interactive influence of winter and/or spring wheat genetic background on haploid induction parameters and trait correlation was studied by hybridizing five elite and diverse genotypes each of winter and spring wheat and their F₁s (winter × winter, spring × spring, and winter × spring, generated in a diallel design excluding reciprocals) with a single genotype of maize. Data were recorded with respect to per cent seed formation, embryo formation, and regeneration. High genetic variability was present among the wheat genotypes (parents + F₁s) for the three haploid induction parameters. Significant differences were obtained within and between different groups viz., spring wheats, winter wheats, spring × spring wheats, winter × winter wheats, and winter × spring wheats with respect to the three haploid induction parameters based on ANOVA. The winter genotypes (winter parents and winter × winter wheat hybrids) responded better than the spring groups (spring wheat parents, spring × spring and winter × spring wheat hybrids) with respect to embryo formation and winter × spring wheat hybrids yielded significantly the highest numbers of regenerants. Correlation studies amongst the haploid induction parameters indicated that the genes controlling seed formation and haploid plantlet regeneration are negatively correlated when the genetic backgrounds of both ecotypes are combined in winter × spring hybrids. Haploid embryo formation had no association with seed formation and regeneration in all genetic backgrounds, suggesting independent inheritance.     

High Frequencies of Fertilization and Haploid Seedling Production in Crosses Between Commercial Hexaploid Wheat Varieties and Maize

Nineteen commercial hexaploid wheat varieties were crossed with the maize F₁ hybrid ‘Seneca 60’. Fertilization frequencies ranged from 32.1 % to 47.5 % of pollinated florets (mean 39.5 %) in the 14 winter wheat varieties and from 40.7 % to 51.4 % (mean 47.8 %) in the five spring wheat varieties. In some cases only an endosperm was formed and the frequencies of embryo formation were therefore slightly lower, being 28.2 % to 45.9 % (mean 36.4 %) for winter wheats and 39.8 % to 48.6 % (mean 45.1 %) for spring wheats. Mean values were significantly higher in the spring wheats but no significant variation was found between varieties within the spring or winter categories. In the five spring wheats the mean yield of embryos, and hence the potential yield of haploid plants, was 3.4-fold higher than with the tetraploid Hordeum bulbosum clone PB179. For the 14 winter wheats the figure was 10.9-fold higher. These differences were highly significant (p < 0.001) in all varieties. A single 2,4-D treatment given to spikes one day after pollination with maize enabled embryos to be recovered from all 19 varieties. A total of 311 embryos were recovered from 950 florets (an average of 7.3 embryos per spike) of which 191 germinated, giving an average yield of one haploid plant for every 5.0 florets pollinated (4.4 haploid plants per spike).     

Sexual hybridization between Capsella bursa‐pastoris (L.) Medik (♀) and Camelina sativa (L.) Crantz (♂) (Brassicaceae)

The development of transgenic oilseed Camelina sativa (2n = 40) and the potential for hybridization with its weedy relative Capsella bursa‐pastoris (2n = 36) necessitates a careful evaluation of the reproductive compatibility between the species. Here, we conducted over 1800 crosses (emasculation and manual pollination) to examine the ability of 10 Canadian C. bursa‐pastoris (♀) accessions to hybridize with five accessions of C. sativa (♂). Seven hybrids were confirmed among 586 putative hybrids screened with species‐specific markers, indicating a hybridization rate of 1.5 hybrids per 10 000 ovules pollinated. All seven hybrids had intermediate DNA content compared to their parents, were morphologically distinct, had low (1.9%) pollen fertility and failed to produce selfed or backcrossed seed. Given the abundance of C. bursa‐pastoris along field margins, hybrids will likely be generated in the wild, but they will be unable to establish lineages unless fertility is restored. The large number of crosses and the diversity captured by the use of multiple accessions resulted in strong statistical power and a high degree of confidence in the estimated hybridization rate.     

Identification and characterization of sources for photo‐ and thermo‐insensitivity in Vigna species

Cultivation of the same varieties of mungbean and blackgram across different seasons and locations is constrained by their photo‐ and thermo‐sensitive behaviour. Developing insensitive genotypes, which can fit well across all seasons, requires robust donors which would provide genes imparting this trait. This study was undertaken to identify such donors in the Vigna species. Forty‐eight accessions belonging to 13 Vigna species and eight released cultivars were evaluated under natural field conditions. Among these, two accessions, viz. V. umbellata (IC251442) and V. glabrescens (IC251372) were found photo‐ and thermo‐insensitive as these were able to flower and set pods at temperatures as high as 43.9°C and as low as 2.7°C. Pollen viability studies indicated viable pollen (>75% at 2.7°C and >85% at 41.9°C) and normal pollen tube growth at both the extremes of temperature. The identified V. glabrescens accession has long, constricted pods and dark green, mottled seeds while V. umbellata has smooth, curved pods and shining, oval, large seeds. Both these accessions can be utilized in developing photo–thermo insensitive genotypes in cultivated Vigna species.     

Improved Techniques for Haploid Production in Wheat using Chromosome Elimination

Wheat × maize and wheat × pearl millet crosses have proved efficient for haploid production using various genotypes of wheat; 22 and 27 % of florets produced embryos. In favourable conditions 6—9 haploid plants per spike were produced. The following simplifications or improvements in technique are recommended: 1. Only a single treatment with an aqueous solution of dicamba or 2,4-D (50–100 ppm) for embryo stimulation in vivo; 2. Application by spraying or dipping the spikes; 3. Application time two to four days after pollination; 4. Embryo rescue 15 to 18 days after pollination; 5. Crosses without emasculation are possible if pollination occurs 1–2 days before anthesis. More than 450 haploids and some doubled haploid (DH) lines (after colchicine treatment in vitro) were produced using these methods. No hybrid plants, chromosome additions or substitutions were found.     

Genetic variation for resistance to Fusarium head blight in bread wheat

Summary During a four year period, a total of 258 winter and spring wheat genotypes were evaluated for resistance to head blight after inoculation with Fusarium culmorum strain IPO 39-01. It was concluded that genetic variation for resistance is very large. Spring wheat genotypes which had been reported to be resistant to head blight caused by Fusarium graminearum were also resistant to F. culmorum. The resistant germplasm was divided into three gene pools: winter wheats from Eastern Europe, spring wheats from China/Japan and spring wheats from Brazil. In 32 winter wheat genotypes in 1987, and 54 winter wheat genotypes in 1989, the percentage yield reduction depended on the square root of percentage head blight with an average regression coefficient of 6.6. Heritability estimates indicated that for selection for Fusarium head blight resistance, visually assessed head blight was a better selection criterion than yield reduction.     

Winter × spring wheat hybridization - a promising avenue for yield enhancement

Introgression of the winter gene pool into spring wheat is being considered as one of the strategies to break through the yield plateau. However, little information is available on the combining ability of these two important but distinct groups of wheats in Indian conditions. Therefore, the present study was undertaken to determine the combining ability and gene action of yield and yield attributes in winter × spring wheat crosses. Seventy F₁ progenies developed by 14 winter and five spring wheat lines using a line × tester design were evaluated, along with their parents, for yield and yield attributes in a randomized complete block design under field conditions. The mean squares for all the characters studied showed highly significant differences. The mean squares due to female × male interactions were significant for all the characters studied except for grains per ear and grain weight per ear. Additive genetic effects were found to play a key role in controlling the expression of days to heading, plant height and spikelets per ear.‘MV 19’ and ‘Stepniak’/‘Karvuna’ among winter and ‘PBW 65’ among spring wheats were good general combiners for most of the yield attributes studied. The estimates for specific combining ability effects suggested that, although general combining ability (GCA) effects of most winter wheats are either average or poor, their combination can give desirable genotypes with spring wheat parents possessing a high GCA.     

Iron and zinc grain density in common wheat grown in Central Asia

Sixty-six spring and winter common wheat genotypes from Central Asian breeding programs were evaluated for grain concentrations of iron (Fe) and zinc (Zn). Iron showed large variation among genotypes, ranging from 25 mg kg⁻¹ to 56 mg kg⁻¹ (mean 38 mg kg⁻¹). Similarly, Zn concentration varied among genotypes, ranging between 20 mg kg⁻¹ and 39 mg kg⁻¹ (mean 28 mg kg⁻¹). Spring wheat cultivars possessed higher Fe-grain concentrations than winter wheats. By contrast, winter wheats showed higher Zn-grain concentrations than spring genotypes. Within spring wheat, a strongly significant positive correlation was found between Fe and Zn. Grain protein content was also significantly (P < 0.001) correlated with grain Zn and Fe content. There were strong significantly negative correlations between Fe and plant height, and Fe and glutenin content. Similar correlation coefficients were found for Zn. In winter wheat, significant positive correlations were found between Fe and Zn, and between Zn and sulfur (S). Manganese (Mn) and phosphorus (P) were negatively correlated with both Fe and Zn. The additive main effects and multiplicative interactions (AMMI) analysis of genotype × environment interactions for grain Fe and Zn concentrations showed that genotype effects largely controlled Fe concentration, whereas Zn concentration was almost totally dependent on location effects. Spring wheat genotypes Lutescens 574, and Eritrospermum 78; and winter wheat genotypes Navruz, NA160/HEINEVII/BUC/3/F59.71//GHK, Tacika, DUCULA//VEE/MYNA, and JUP/4/CLLF/3/II14.53/ODIN//CI13431/WA00477, are promising materials for increasing Fe and Zn concentrations in the grain, as well as enhancing the concentration of promoters of Zn bioavailability, such as S-containing amino acids.     

Interspecific hybridization in Gossypium L.: characterization of progenies with different ploidy‐confirmed multigenomic backgrounds

Cytological and molecular investigations were undertaken for parent and progeny derived from a trispecific line [2(Gossypium arboreum × G. anomalum) × G. hirsutum var. BWR], which was crossed with G. hirsutum var. JLH168. Cytomorphological analysis of the F₁ (G. arboreum × G. anomalum), its amphidiploid and progeny from trispecies hybrid showed distorted ploidy segregation with monovalents to hexavalents and high intergenomic (small A₂ and large B₁) allosynthetic chromosome pairing. Microsatellite analysis identified three fragments associated with G. arboreum and G. anomalum and six fragments associated with G. hirsutum in derivates of the trispecies line × G. hirsutum var. JLH168. Inter‐Retrotransposon Amplified Polymorphism (IRAP) analysis revealed fragments of G. arboreum and G. anomalum, only in F₁ and amphidiploid. Chromosomal association and microsatellite analysis of three progeny genotypes (i.e. haploid, hexaploid and tetraploid no. 1) confirmed that they share multigenomic background from the three cotton species (A₂, A_hD_h and B₁ genome). The interspecific hybrid cotton genotypes studied are likely to be useful for the introgression of genes from diploid species to commercial upland cultivars.     

The site of action of crossability genes (Kr 1, Kr 2) between Triticum and Secale. II. Proportion of pistils containing pollen tubes and effects of alternate pollination on seed set

Summary Different wheat genotypes (T. aestivum) were crossed with rye to ascertain the site or sites of manifestation of the crossability genes, Kr ₁ and Kr ₂. By using fluorescence microscopy, it was found that the order of increasing proportion of wheat micropyles containing pollen tubes is strongly correlated with the levels of crossability with rye. High crossable genotypes have more micropyles containing pollen tubes than those of the low crossable ones. Most of the inhibition or retardation of pollen tubes occurred between the style base and top of the embryo sac, expecially with the low crossable genotypes where both Kr-genes are present. The results also indicate that Kr ₁ is a greater inhibitor than Kr ₂. Seed set is also highly correlated with the number or proportion of micropyles having pollen tubes. Alternate pollinations seem to support the view that rye pollen tubes do not reach the micropyles of the low crossable genotypes, and hence when repollinated with wheat selfed seeds are produced.     

Distribution and allelic expressivity of genes for hybrid necrosis in some elite winter and spring wheat ecotypes

The distribution and allelic expressivity of hybrid necrosis genes (Ne ₁ and Ne ₂) were studied in 21 winter (mostly exotic) and 43 spring type elite wheat genotypes, by crossing them with two known testers, C 306 (Ne ₁-carrier) and HD 2380 (Ne ₂-carrier).Ne ₁ gene was present in one north-west Himalayan winter wheat landrace, Shoure Local, but absent in the other winter as well as spring wheats. Ne ₂ gene was prevalent to a much lower extent in the exotic winter wheat germplasm (31.57%) as compared to the recently developed Indian and Mexican spring wheat semidwarfs (69.80%). This may suggest that breeders have tried to preclude hybrid necrosis by selecting for non-carrier genotypes in the development of exotic winter wheats in contrast to the situation in spring wheats. Based on the degree of expression of hybrid necrosis genes in the F₁ hybrids, the carrier genotypes were characterized with respect to the allelic strength of the hybrid necrosis genes. The 27 non-carrier genotypes of the two ecotypes identified in the present study have a greater potential use in future hybridization programmes so as to overcome the problem of hybrid necrosis. This revised version was published online in July 2006 with corrections to the Cover Date.     

Inheritance of resistance to wheat midge, Sitodiplosis mosellana, in spring wheat

Inheritance of resistance to a wheat midge, Sitodiplosis mosellana (Géhin), was investigated in spring wheats derived from nine resistant winter wheat cultivars. F₁ hybrids were obtained from crosses between resistant winter wheats and susceptible spring wheats, and used to generate doubled haploid populations. These populations segregated in a ratio of 1:1 resistant to susceptible, indicating that a single gene confers the resistance. The F₂ progeny from an intercross among spring wheats derived from the nine resistance sources did not segregate for resistance. Therefore, the same gene confers resistance in all nine sources of resistance, although other genes probably affect expression because the level of resistance varied among lines. Heterozygous plants from five crosses between diverse susceptible and resistant spring wheat parents all showed intermediate levels of response, indicating that resistance is partly dominant. Susceptible plants were reliably discriminated from heterozygous or homozygous resistant ones in laboratory tests, based on the survival and development of wheat midge larvae on one or two spikes. This powerful resistance gene, designated Sm1, is simply inherited and can be incorporated readily into breeding programmes for spring or winter wheat. However, the use of this gene by itself may lead to the evolution of a virulent population, once a resistant cultivar is widely grown.     

Crossability and embryo rescue enhancement in wide crosses between wheat and three Agropyron species

Summary Soft winter wheat lines were crossed with Agropyron intermedium, A. elongatum and A. trichophorum using pollen from single plants of Agropyron spp. to pollinate wheat spikes. Not only species but also individual plants within varieties of Agropyron species differed in percent seed set with a wheat genotype. In two arrays of crosses between two phenotypically different plants of A. elongatum and nine wheat lines, one Agropyron plant gave higher seed set (overall=27.1%) than the other (overall=3.7%). The differences were significant in seven of the nine cross combinations. Results are consistent with the hypothesis that these two plants differ in their crossability as pollen parents with wheat, and suggest the possibility of occurrence of crossability genes in wheatgrasses. The success rate of hybrid embryo rescue was higher (87.5%) with cold treatment (4°C) than without cold treatment (75.0%) of excised embryos on culture media. Results underscore the significance of genotype of the alien species for crossing with low crossable wheats, and of the physical factors for improving embryo rescue in wide crosses.Contribution No. 11,825, Purdue Agric. Exp. Stn., West Lafayette, Indiana 47907, USA. The research was supported in part by Public Varieties of Indiana.     

Influence of flower and flowering characteristics on seed yield in diploid and tetraploid red clover

Tetraploid red clover (Trifolium pratense L.) often produces insufficient seed to be economically interesting. Numerous studies have identified poor pollination due to long corolla tubes as its cause, but other studies disagree. Therefore, we investigated seed yield, corolla tube length and flowering traits in 244 plants from five diploid and five tetraploid cultivars during 2 years. Tetraploids produced fewer seeds, fewer flower heads and fewer seeds per head, but an equal number of flowers per head when compared to diploids. Although corolla tubes were longer in tetraploid than in diploid red clover (P < 0.001), no correlation between corolla tube length and seed yield was detected. Therefore, the corolla tube length is likely neither the main nor only cause of low seed yield in tetraploid red clover. Instead, three putative causes for low seed yield were identified: tetraploids produced less inflorescences per plant, possessed a lower degree of determinacy and had markedly less seeds per head when compared to diploids. Possible causes for such differences include lower pollen viability and higher rates of embryo abortion.     

Reproductive compatibility studies between wild and cultivated strawberries (Fragaria × ananassa) to obtain “bridge species” for breeding programmes

The Fragaria genus has become relevant due to worldwide economic importance of the octoploid hybrid F. × ananassa. Interspecific hybridizations are widely employed in breeding programmes to introduce useful characteristics from wild species to cultivated varieties, although many reproductive barriers prevent gene flow between species. Hybrid genotypes that can act as a bridge between related wild species and cultivated strawberry could be useful to overcome those barriers. The aim of this work was to obtain interspecific hybrids of Fragaria and to analyse possible reproductive barriers. In the Banco de Germoplasma de Frutilla‐UNT, crossings were carried out among wild species of Fragaria with different ploidy levels and cultivated strawberry. To confirm hybrid condition of genotypes obtained, histological techniques and microsatellite markers were used. When wild species were crossed with cultivated strawberry in both directions, there was low production of achenes, while in crosses between wild species of Fragaria genus, achene production was very high. The percentage of germination of achenes was high when crossed species had the same ploidy level, and very low or null when they presented different ploidy levels. Although pre‐zygotic incompatibility associated to the degree of domestication and postzygotic barriers related to different ploidy levels of the progenitors were detected, new hybrid genotypes of Fragaria were obtained. These new hybrids could be used as “bridge species” in breeding programmes, since preliminary results showed no incompatibility barriers when they were crossed with Duchesnea indica.     

Chromosome pairing and fertility of interspecific hybrids between Trifolium repens L. and T. occidentale Coombe

Trifolium occidentale is a diploid wild relative with the potential to improve the drought and salt tolerances of white clover (Trifolium repens). Previous work has shown that it is possible to efficiently produce large T. repens × T. occidentale breeding populations using colchicine‐doubled (4×) T. occidentale. For effective introgression (backcross) breeding, it is also essential that interspecific chromosome pairing and recombination occur. In this study, it was apparent that chromosome pairing was occurring not only between T. occidentale and T. repens subgenomes, but also between the ancestral subgenomes of T. repens. Thus, interspecific hybridization has the potential for major genome recombination and opens the way for introgression of traits from T. occidentale into white clover.     

Molecular cytogenetic characterization of two partial wheat Elytrigia elongata amphiploids resistant to powdery mildew

Two new partial amphiploids SN20 and SN122, derived from crosses between Elytrigia elongata and common wheat Yannong15 and Shannongfu63, were found resistant to powdery mildew, a serious fungal wheat disease. Cytogenetic observations showed that each SN20 and SN122 plant had 56 chromosomes. The chromosomes in most pollen mother cells formed approximately 28 bivalents, thereby showing a high degree of cytogenetic stability. Genomic in situ hybridization using St‐genomic DNA from Pseudoroegneria strigosa as a probe and ABD‐genomic DNA from Chinese Spring wheat as a blocker demonstrated that the genomic formulas for the alien chromosomes in SN20 and SN122 were 2 St + 10 J^S + 2 J and 2 St + 8 J^S + 4 J, respectively. These wheat E. elongata partial amphiploids represent a potential novel source of resistance to powdery mildew for wheat breeding.     

Genetic Variability for Haploid Production in Crosses Between Tetraploid and Hexaploid Wheats with Maize

Crosses were made between 14 wheat genotypes (11 tetraploid, 3 hexaploid) and a single Fl hybrid of maize that was used as the male parent. The experimental design consisted of randomized blocks with three replications. Plants were grown under controlled greenhouse conditions (day length 16 h and temperature 25 °C/15 °C, day/night). To enhance embryo survival, 2, 4-D treatment (10 mg/1) was applied to spikes 24 h after pollination with maize. Embryos were recovered from all tetraploid and hexaploid wheats at a rate of 2.09 to 26.76 per 100 pollinated florets. Haploid and doubled haploid plants were obtained from all hexaploid genotypes (T. aestivum) and from 5 of 11 tetraploid genotypes (T. turgidum var.). The most important point of these experiments was the ability to produce haploid plants from tetraploid wheat for two reasons: firstly, anther culture cannot be applied in tetraploid wheat (T. turgidum var.) due to the inefficiency of embryo formation and the high proportion of albino plants. Secondly, to date, crosses between tetraploid wheat and maize have resulted in embryo formation, but not in haploid plants.     

Hybridization between Camelina sativa (L.) Crantz (false flax) and North American Camelina species

The potential for gene flow between Camelina sativa, a promising edible and industrial oil crop in Canada, and its wild North American relatives C. alyssum, C. microcarpa and C. rumelica subsp. rumelica, was investigated. Sequence analysis of the nuclear ribosomal internal transcribed spacer region (ITS) was used to differentiate Camelina species and/or accession‐specific nucleotide markers and identify interspecific F₁ hybrids. ITS analysis identified hybrids in progeny of C. alyssum × C. sativa, C. microcarpa accession 36010 × C. sativa and C. sativa × C. rumelica subsp. rumelica. Seed set on C. alyssum × C. sativa F₁ and F₂ progeny was similar to the parents; few seeds were produced on hybrid progeny of C. microcarpa accession 36010 × C. sativa and C. sativa × C. rumelica subsp. rumelica. The study provided evidence that should the species have sympatric distributions and overlapping flowering periods, gene flow between C. sativa and its wild North American relatives is possible and that it would most likely occur with C. alyssum given the high fertility of the F₁ hybrids recovered.