Durum wheat × maize crosses for haploid wheat production: Influence of parental genotypes and various experimental factors

To improve haploid plant production in durum wheat, the haplomethod involving intergeneric crossing with maize followed by embryo rescue was used. The influence of parental genotypes and various experimental factors were studied. Ten cultivars of Triticum turgidum ssp. durum (female parent) were crossed with eight genotypes of Zea mays. After pollination, plant stems were either maintained in situ or cut near the base and kept in a 2,4‐dichlorophenoxyacetic acid (2,4‐D)‐sucrose solution. Ten to 18 days after pollination, embryos were excised from developed ovaries and cultured on one of MS, MS/2, or B5 media. Haploid embryos and plants were obtained (78 green haploid plants regenerated in 0 year). The wheat genotype was significant for ovary development, embryo and plant formation, whereas the maize genotype was significant only for embryo formation. Detailed results of all crosses showed the best crossing partner for each wheat genotype. Cutting the plant stems after pollination gave better results than maintaining them in situ. The optimal stage for embryo rescue was 14 days and B5 and MS/2 media were more efficient than MS for embryo culture.     

Influence of selected wheat and rye genotypes on the direct synthesis of hexaploid triticale

Summary In the synthesis of primary hexaploid triticale, a cross-incompatibility barrier exists when tetraploid wheat (4X) is crossed with diploid (2X) rye. Fertilization may occur, however, abnormal endosperm development usually leads to premature embryo death. Four selected tetraploid wheat lines were crossed as females with seven open-pollinated rye lines and the resulting embryos were rescued in vitro 13–16 days after pollination. The wheat genotypes showed a major influence on crossability (seed set), embryo development and plant recovery. The highest efficiency of amphihaploid plant recovery (18.3 plants per 100 pollinated florets) was obtained from one 4X wheat line originally selected from the cross T. carthlicum × T. dicoccoides. Some of the 3X amphihaploid plants (ABR) derived from two wheat lines showed relatively high level of partial fertility presumably as a result of meiotic restitution. Correlation analysis showed that crossability (seed set), normal hybrid embryo development in vivo and embryo culturability were independent of each other.     

Production of haploids in bread wheat, durum wheat and hexaploid triticale crossed with pearl millet

Pearl millet is an efficient alternative to maize as a pollen source for haploid production in bread wheat. To compare haploid production frequencies in other Triticeae species, the crossabilities of two genotypes each of bread wheat, durum wheat and hexaploid triticale with four pearl millet genotypes and a maize control were examined. Embryos were obtained from crosses of all three species with both pearl millet and maize. However, significant differences in crossability were found among the three species (10.5–79.8% seed development and 1.4–15.8% embryo formation), as well as among genotypes of durum wheat (7.2–23.7% and 2.1–6.4%) and hexaploid triticale (0.3–20.6% and 0.1–2.7%). Crossability of bread wheat with pearl millet was relatively high. Haploid plants were regenerated from crosses of all three species with pearl millet. As in the case of maize crosses, low crossabilities of durum wheat and hexaploid triticale with pearl millet can be attributed to the absence of D-genome chromosomes.     

Parental effects on crossability,embryo differentiation and plantlet recovery in wheat x rye hybrids

Summary One hexaploid wheat cultivar (Triticum aestivem) and two tetraploid wheat lines (T. durum) were crossed with seventeen inbred lines of rye (Secale cereale). Seed set, degree of hybrid embryo differentiation at the time of excision for in vitro culture and recovery of amphihaploid plantlets from various embryo categories were studied. Degree of embryo differentiation was predominantly determined by maternal wheats, paternal rye genotypes appearing to be of minor importance. T. aestivum x rye hybrid embryos were superior to those produced from T. durum for degree of differentiation. The proportion of plantlets developing from differentiated embryos was high for all wheat parents, whereas undifferentiated embryos were mostly unsuitable for plantlet production. The results revealed that cross-incompatibility in hexaploid wheat x rye crosses was due to failure of fertilization, while in tetraploid wheat x rye crosses it was caused by lack of embryo differentiation. Correlation analyses showed that seed set provided a criterion to predict the amphihaploid plantlets to be expected from a particular wheat x rye combination.     

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).     

Endosperm peroxidase electrophoresis patterns to distinguish tetraploid from hexaploid wheats

Summary A simple method is proposed to distinguish hexaploid (Triticum aestivum L.) from tetraploid (Triticum turgidum L., durum wheat) cultivated wheats on the basis of peroxidase isozymes coded by genome D. It can also be used as a first step to detect possible contamination by tetraploid genotype mixtures. The peroxidase patterns of endosperm and of embryo plus scutellum found among 349 entries of a durum wheat world basis collection are shown.     

Resistance to stripe rust in Triticum turgidum,T. tauschii and their synthetic hexaploids

Resistance to stripe rust (caused by Puccinia striiformis Westend.) of 34 Triticum turgidum L. var.durum, 278 T. tauschii, and 267 synthetic hexaploid wheats (T. turgidum x T. tauschii) was evaluated at the seedling stage in the greenhouse and at the adult-plant stage at two field locations. Mexican pathotype 14E14 was used in all studies. Seedling resistance, expressed as low infection type, was present in all three species. One hundred and twenty-eight (46%) accessions of T. tauschii, 8 (23%) of T. turgidum and 31 (12%) of synthetic hexaploid wheats were highly resistant as seedlings. In the field tests, resistance was evaluated by estimating area under disease progress curve (AUDPC). Synthetic hexaploid wheats showed a wide range of variability for disease responses in both greenhouse and field tests, indicating the presence of a number of genes for resistance. In general, genotypes with seedling resistance were also found to be resistant as adult plants. Genotypes, which were susceptible or intermediate as seedlings but resistant as adult plants, were present in both T. turgidum and the synthetic hexaploids. Resistances from either T. turgidum or T. tauschii or both were identified in the synthetic hexaploids in this study. These new sources of resistance could be incorporated into cultivated hexaploid wheats to increase the existing gene pool of resistance to stripe rust.     

Influence of genotype and culture conditions on the production of embryos from anthers of tetraploid wheat (Triticum turgidum)

Summary Anther culture of 10 tetraploid wheat (Triticum turgidum) genotypes and two backcross lines representing a wide range of genetic variation was studied in a randomized block design with three replications. Each replication consisted of 2 pots with 3 plants. The day length was 16h and temperature 25° C/15°C for day/night in a controlled greenhouse where the anther donor plants were grown. Two different treatments were used for anther culture. The first one was potato 2 medium (Chuang et al., 1978) modified by adding 0.5 mg/l glutamine and solidified by gelrite (4g/l) (Henry & De Buyser, 1981). Cultures were incubated in light (15 E m^–2 S^–1) at 26°C at 16h day length. The second medium was described by Fadel & Wenzel (1990), differing from the first by the nature of the sugar (maltose) and consistency of the medium (semiliquid by ficoll). Anther cultures were incubated in the dark at 28°C. The study of about 1300 anthers per genotype and treatment showed that both genotype and treatment affected embryo formation of tetraploid wheat. The backcross lines exhibited significant differences for androgenic abilities when compared to their common parent. Most of the genotypes were medium dependent for androgenesis and revealed significant interactions with the two treatments. Five green plantlets were regenerated and fertile doubled haploid plants were obtained from three out of the 12 studied genotypes.     

The production and behaviour of hexaploid hybrids between Hordeum vulgare and H. Bulbosum

Summary To produce hexaploid (or other polyploid) hybrids, diploid or tetraploid Hordeum vulgare was crossed with hexaploid or octoploid H. bulbosum, and perennial triploid hybrids between the two species were treated with colchicine. The crosses did not yield viable plants: seedset was low, the seed aborted and embryo culture was unsuccessful. The colchicine treatments geve rise to plants in which hexaploid chromosome numbers were observed. At the hexaploid level chromosomal instability occurred, resulting in chromosome elimination.The colchicine-treated triploid hybrids showed in the first years after the treatment better fertility after open flowering than untreated plants, but the level of fertility remained very low. The offspring consisted of haploid, diploid and approximately triploid plants like H. vulgare, tetraploid and approximately tetraploid plants like H. bulbosum, and plants with hybrid morphology and unstable chromosome number, which were highly sterile. Thus the crossing barrier between H. vulgare and H. bulbosum could not be broken down at higher ploidy level.     

The effects of parthenogenesis on wheat embryo formation and haploid production with and without maize pollination

Doubled haploid (DH) lines are important in wheat (Triticum aestivum L.) breeding, and haploids produced via maize pollination precede DH line development. Although maize pollination has proven reliable and broadly applicable to wheat, its success is determined by the wheat and maize genotypes employed. A wheat genotype consisting of nuclear and cytoplasm components predisposing it to parthenogenesis was compared with three other genotypes, each possessing only one or neither component necessary for parthenogenesis. In a glasshouse experiment, each genotype was pollinated with maize and subsequently treated with a2,4-Dichlorophenoxyacetic Acid (2,4-D) solution to determine if parthenogenesis affected embryo formation frequency (EFF)and haploid formation efficiency (HFE). Wheat genotypes were also treated with the2,4-D solution alone to determine if embryos and haploid plants could be produced in vivo without maize pollination. ‘Salmon(K)’, a parthenogenetic genotype consisting of a Salmon 1BL.1RS nucleus in a Ae. kotschyii cytoplasm, had a mean EFF of 32%; whereas, the non-parthenogenetic genotypes had mean EFF calculations ranging from 7 to 21%. Mean HFE for Salmon(K) was not significantly different than the mean HFE for non-parthenogenetic Salmon; however, EFF and HFE calculations for Salmon(K) and Salmon, each with a 1BL.1RS translocation, were generally higher than calculations for genotypes without the translocation. Salmon(K) was the only genotype to produce a 3% or higher EFF and HFE after treatment with 2,4-D alone. Parthenogenesis significantly affected the frequency at which embryos were produced after pollination with maize and the frequency at which embryos and haploid plants were produced after treatment with 2,4-D alone. This revised version was published online in July 2006 with corrections to the Cover Date.     

Crossability of tetraploid and hexaploid wheats with ryes for primary triticale production

Summary Crossability of wheat and rye was investigated during thirteen crop cycles in two contrasting locations to 1) evaluate tetraploid and hexaploid wheat parents in crosses with rye, 2) identify genotypes with high crossability and 3) assess the impact of environment on seed development. The majority of the tetraploid wheats crossed with rye had seed set around 20%, but very low embryo viability. Several wheat genotypes with seed set above 50% were identified. The hexaploid wheats crossed with rye showed poor seed set, but plant recovery was relatively high. The majority of the hexaploid wheats with highest seed set (20–30%) were from China. The results suggest differences in crossability between the rye populations, and wheat species by rye interactions. The crossability of the tetraploid and hexaploid wheats was affected by climate in the two locations.     

Transfer of disease resistance genes from Triticum araraticum to common wheat

The wild tetraploid wheat species Tr$$ (Zhuk) Zhuk Var. araratieum is a source of pest resistance genes for T$$ aesti$$ L. Our objectives were to describe the breeding behaviour of T.arartuititm when backcrossed to common wheat and transfer resistance to leaf rust (caused by Pu$$) and powdery mildew (caused by Blumeria $$wheat. Crosses were made between five wheat genotypes and $$ accessions. Fertifity and chromosome numbers of BC$$_; plants were determined. Resistance to leaf rust was transferred toBC₂ -derived families from 10 different T’ararati$$an accessions. Leaf rust resistance genes in nine T. araratieum accessions can be assigned to at least four loci. Leaf rust resistance transferred from three accessions was inherited in the hexaploid derivatives as a single. $$ gene in each case. Resistance to powdery mildew was also detected in the T. araratie$$ backcross derivatives. Fertile hexaploid derivatives expressing T’araratieum-derived resistance genes can be recovered after two backcrosses to wheat cultivars.     

Wheat embryogenesis and haploid production in wheat × maize hybrids

Summary Embryogenesis was analyzed in wheat × maize hybrids using paraffin sectioning. Embryogenesis in wheat × maize hybrids is different from that in self-pollinated wheat plants. Development of the embryo is not accompanied by the formation of an endosperm. The endosperm nuclei remain free in the cytoplasm, fail to advance into the cellular stage, and degenerate at a later time. The antipodal cells quickly degenerate in the fertilized ovaries of wheat × maize hybrids similar to self-pollinated ovaries. The antipodal cells remain normal in unpollinated ovaries. The pre-embryo will abort if it is allowed to develop on the plant, because of a nutritional shortage in the absence of an endosperm. Therefore, embryo rescue is necessary for haploid production from a wheat × maize hybrids. Haploid polyembryos were obtained from spikelet culture of wheat × maize hybrids. The formation of polyembryos is due to the cleavage of the pre-embryo and the effect of 2,4-D. The frequency of haploid embryo production and plant regeneration is affected significantly by maize genotypes, but not by wheat genotypes. The concentration of 2,4-D affects only the size of the embryo.     

Suppression/expression of resistance to stripe rust in synthetic hexaploid wheat (Triticum turgidum×T. tauschii)

Seventy-four hexaploid wheats, synthesized by either crossing resistantTriticum turgidum L. var.durum with susceptible/intermediateT. tauschii or susceptible/intermediateT. turgidum with resistantT. tauschii, and their parents were evaluated as seedlings in the greenhouse and as adult-plants at two field locations in Mexico for resistance to pathotype 14E14 of stripe (or yellow) rust (caused byPuccinia striiformis Westend). The seedlings of different synthetic hexaploids showed high phenotypic diversity for resistance. However, the resistance level of only 15 of the 74 synthetic hexaploid wheats were similar to the low infection types of the respective donor parents. The remaining synthetic wheats displayed either intermediate or high infection types. A similar result was also obtained in field tests, where only 18 synthetic hexaploids were resistant as adult-plants. In general, genotypes with seedling resistance were also resistant as adult-plants. A few synthetic hexaploids, which displayed intermediate or susceptible infection types as seedlings were resistant as adult-plants, indicating that additional genes for adult-plant resistance were also present. The fact that resistance of some donor parents was not expressed, or only partially expressed, in a synthetic hexaploid background suggests the presence of suppressor genes in the both the A or B, and D genomes ofT. turgidum andT. tauschii, respectively. The resistance of a donor parent was expressed in a synthetic hexaploid only if the corresponding suppressor was absent in the second parent. Moreover, the suppressors appeared to be resistance gene specific.     

Variation of high-molecular-weight glutenin subunits amongst cultivars of Triticum turgidum L. from Portugal

Summary Variation in high-molecular-weight (HMW) glutenin subunit composition amongst 63 varieties of Triticum turgidum L. from Portugal was investigated using SDS-PAGE. A total of thirteen Glu-A1 and Glu-B1 alleles were identified, and three of them were found to be different from those previously described in the literature. A number of the tetraploid wheats examined contained subunits known to have a beneficial effect of the bread-making properties of T.aestivum. Camara, a tetraploid wheat cultivar carrying a 1D/1B chromosomal substitution, is proposed as a bridge for the transfer of Glu-D1 alleles and of other protein fractions controlled by the 1D chromosomes (Gli-D1 locus) from hexaploid to tetraploid wheat.     

Growth and senescence characteristics associated with tolerance of wheat-alien amphiploids to high temperature under controlled conditions

Tolerance of wheat (Triticum aestivumL.) to high temperature might be improved by introducing alien genes from amphiploids. Our objectives were to determine responses of synthetic hexaploid and octaploid amphiploid wheats to high temperature and evaluate their potential usefulness for developing improved cultivars. Thirty synthetic hexaploids from durum wheat (T. turgidum L.) × Aegilops tauschii Cos. Accessions and four octaploid amphiploids from Chinese Spring wheat × different grasses were grown at 20/15 and 30/25 ^°C day/night during maturation. Tolerance was ascertained by two measures of senescence, leaf chlorophyll content and grain filling duration, plus grain yield and its components. Leaf chlorophyll was measured after 10 and 15 days of treatment, and grain yield was determined at maturity to calculate the heat susceptibility index(HSI), a gauge of the reduction in yield at high temperature of each line relative to all other lines. Chlorophyll content, grain filling duration, yield, and kernel weight were highly negatively correlated with HIS of the hexaploid amphiploids at30/25 ^°C, but grain yield was positively correlated with HSI at20/15 ^°C. The hexaploid lines might be useful for improving wheat for regions where stress from high temperature occurs frequently. Chlorophyll content and grain filling duration also were highly negatively correlated with HSI of the octaploid lines, but they would be less directly useful for improving wheat because the kernel number was reduced greatly due to unbalanced meiotic chromosomal segregation. This revised version was published online in August 2006 with corrections to the Cover Date.     

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.     

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.     

Variation of B subunits of glutenin in durum,wild and less-widely cultivated tetraploid wheats*

Three major types of B subunits of glutenin patterns were detected among 240 durum wheat lines collected from eleven countries by the one-step one-dimensional SDS-PAGE procedure. Most commercial durum lines had the LMW-2 type while extensive variation of other banding patterns was found for lines particularly from North African and Mediterranean region. A total of 281 wild and less-cultivated tetraploid wheat lines (var. dicoczoides, Triticum dicoccum and T. polonicum) were also studied for the variation of low molecular weight glutenin subunits and extensive variation was found. The complexity of banding patterns observed among the T. turgidum species indicate a rich source of glutenin variation with a potential value to be used for quality improvement of cultivated wheat when their functional properties have been tested.     

The Grain Quality of Landraces of Wheat as Compared with Modern Cultivars

Landraces of tetraploid and hexaploid wheat from the Northern Negev in Israel were evaluated over two years for their grain quality attributes. Twenty-one populations of tetraploid wheat (Triticum durum, represented by 56 accessions) and 8 populations of hexaploid wheat (T. aestivum and T. compactum, represented by 13 accessions) were compared with 3 and 4 modern improved Israeli cultivars of tetraploid and hexaploid reheat, respectively. This comparison allowed to estimate the progress made in the improvement of gram quality m present-day cultivars. Except for grain protein content in the hexaploids, significant variation was revealed among landraces in kernel weight, protein content (tetraploids), sedimentation, mixograph score and carotin content. The best modern cultivars were comparable to the best landraces in kernel weight and carotin content (tetraploids), indicating that modern wheat breeding maximized kernel weight and carotin content, as compared with the tested landraces. Gram protein content and/or quality was not maximized in modern cultivars and its improvement was deemed possible by introgression from the best landraces in this respect.