Factors affecting haploid production in wheat using the Hordeum bulbosum system. 3. Post-fertilization effects on embryo survival

Summary An investigation was made of factors affecting the frequency of development of fertilized ovules following wheat x Hordeum bulbosum hybridizations. Seed survival, a term used here to represent the relationship between seed set and fertilization, was shown to vary between the wheat genotypes tested, but was unrelated to the crossability genes present within the wheat genotype. A high seed survival was obtained in crosses involving the variety TH3929 and the single chromosome substitution line, Chinese Spring (Hope 5B). Chinese Spring showed a poor seed survival. Some influence of the H. bulbosum genotype on seed survival was detected. The two environmental factors studied, namely the post-pollination application of gibberellic acid and the ambient temperature, were both shown to affect seed survival. The application of gibberellic acid immediately after pollination or 1 and 2 days after pollination improved seed survival, although the extent of the response was dependent upon the number and timing of the application(s). A genotype-dependent response to the ambient temperature was observed, TH3929 showing a slight but insignificant reduction and Chinese Spring a significant improvement in seed survival with an increase in temperature from 20°C to 26°C. The physiological processes which may be influenced by GA and the ambient temperature and may thereby affect seed survival are discussed.     

Factors affecting haploid production in wheat using the Hordeum bulbosum system. 2. The effect of the timing of pollination

Summary The effect of manipulating the timing of pollination on the fertilization frequency in wheat × tetraploid Hordeum bulbosum hybridizations was investigated as a possible means of overcoming varietal incompatibility in this cross. Pre- and post-anthesis pollinations were made on previously identified highly crossable and poorly crossable wheat genotypes and the stage of development of the individual florets at pollination was expressed in days from anthesis units.In both crossable and non-crossable wheat genotypes, a significant negative linear relationship was observed between floret age and fertilization frequency. The magnitude of the response of early pollination was dependent on the number of alleles for non-crossability present at the Kr loci. The non-crossable cultivar Highbury, possessing more than one allele for incompatibility, showed a slight response. The greatest response was shown by the highly crossable cultivar Chinese Spring, possessing alleles for crossability at all Kr loci. The single chromosome substitution line, Chinese Spring (Hope 5B), possessing the most potent allele for non-crossability, Kr ₁, showed an intermediate response. The H. bulbosum genotype had no effect on the magnitude of this response, influencing only the mean fertilization frequency.     

Doubled haploid production in winter wheat and triticale genotypes,using the Hordeum bulbosum system

Summary An attempt was made to produce doubled haploids on 16 winter wheat and six spring and winter triticale genotypes thought to carry genes for interspecific incompatibility. The potential for haploid production was maximized by the use of Hordeum bulbosum genotypes selected for high crossability on crossable wheat genotypes, the use of two post-pollination applications of gibberellic acid and by the pollination of immature florets.A low frequency of seed was set on both the wheat and the triticale genotypes, having mean seed sets of 0.20 per cent and 0.27 per cent respectively. Although the frequency of embryos (seed quality) was high, doubled haploid production was further limited by poor embryo differentiation and regeneration. Haploid plantlets were obtained from the wheat cultivars Moulin and Renard, although successful chromosome doubling and doubled haploid production was achieved in Moulin only.     

The crossing of common wheat (Triticum aestivum L.) with cultivated rye (Secale cereale L.). I. crossability,pollen grain germination and pollen tube growth

Summary The crossing of common wheat (Triticum aestivum L.) with rye (Secale cereale L.), and especially the action of the crossability genes of wheat, was studied using the readily crossable wheat cv Chinese Spring (genotype kr ₁ kr ₁ kr ₂ kr ₂ >), the poorly crossable wheat cv Hope (genotype Kr ₁ Kr ₁ Kr ₂ Kr ₂ ), as well as the disomic substitution line of chromosome 5B of Hope into Chinese Spring (CS/Hope 5B, genotype Kr ₁ Kr ₁ kr ₂ kr ₂ ). By comparing crossability and actual fertilization, the poor crossability with rye of both cv Hope and the CS/Hope 5B substitution line was shown to result from absence of fertilization. Studies of pollen grain germination and pollen tube growth showed that the dominant alleles of the crossability genes manifested themselves through retardation and eventually inhibition of pollen tube growth at the style base and in the ovary wall. In Hope the growth of all pollen tubes was inhibited, whereas in CS/Hope 5B rarely fertilization was achieved. The recessive alleles of the crossability genes do not seem to have an influence on the growth of rye pollen tubes in wheat pistils.     

The site of action of the crossability genes (Kr 1, Kr 2) between Triticum and Secale. I. Pollen germination,pollen tube growth,and number of pollen tubes

Summary Seven genotypes of common wheat (Triticum aestivum L.) were crossed with rye (Secale cereale L.) in order to find the site or sites of action of the crossability genes, Kr ₁ and Kr ₂, of wheat. The data obtained, by fluorescence microscopy, were compared to the controls (wheat x wheat). The results indicate that the crossability genes have little effect on pollen germination and on the time taken for the pollen tubes to reach the microphyle, irrespective of their crossabilities with rye. The number of pollen tubes reaching the microphyle is, however, affected by the Kr-genes, as high crossable genotypes have more pollen tubes than the low crossable ones. There was a high correlation between the mean number of pollen tubes at the micropyle with seed set, which also reflects the crossability. The Kr-genes seem to manifest themselves in the retardation and inhibition of pollen tube growth between the style base and the top of the embryo sac, where the effect is most distinct in the low crossable genotypes.     

Temperature effects on seed germination and expression of seed dormancy in wheat

Because preharvest sprouting decreases quantity and quality of wheat grain, researchers need effective protocols to assess response to preharvest sprouting conditions. The aim of this study was to determine which temperature gives the greatest difference in seed germination and expression of seed dormancy in 10 spring wheat genotypes. The genotypes were grown in the field near Swift Current, Saskatchewan in 2000 in a randomized complete block with four replicates. Seed samples were harvested at approximately 25% moisture content (wet weight basis) and dried to 12% moisture content with minimal after-ripening. Germination was under controlled environment at temperatures of 10, 15, 20 and 30 °C in darkness. A weighted germination index (WGI) was calculated. The analysis of WGI, for each temperature, showed highly significant (p ≤ 0.01) genotype effects on germination. Most genotypes decreased in WGI (increased dormancy) as temperature was increased from 10 to 30 °C. The greatest differences in seed germination tended to be at 15 °C and 20 °C. The level of seed dormancy depended on the genotype and germination temperature. This revised version was published online in August 2006 with corrections to the Cover Date.     

Effects of desiccation and a change in temperature on germination of immature grains of wheat (Triticum aestivum L.)

A study was made of the effects of desiccation and a change in temperature on the germination of wheat grains harvested 20 days after anthesis. When the germination test was performed immediately after harvesting, germination percentages ranged from 5.2% to 10.7%. Germination percentages increased to 48.2% to 90.3% after grain had been desiccated at 20 °C and then hydrated at 10 °C. This increase occurred even if grains had been desiccated in an atmosphere of high relative humidity. The germination percentage of non-desiccated grains depended on the germination temperature. When the pericarp and testa were removed from embryos, the germination percentage of grains incubated at 20 °C and then at 10 °C was higher than that of grains incubated at 10 °C. In general, a low germination temperature is believed to be effective in breaking dormancy of wheat grains. However, a change in temperature stimulated germination to a greater extent than a constant low temperature. The germination percentage of 5-day cycle alternating temperature was greater than that with 1-day, 2-day and 10-day cycles. Although the germination of immature wheat grains was stimulated by both high and low germination temperatures, it is likely that cycles shorter and, also, longer than a critical period induce limited germination. Loss of dormancy commonly occurs when development of wheat grains proceeds at a high temperature, with imbibition at a low temperature. However, germination ability of non-desiccated immature wheat grain was enhanced by a change in temperature during germination. This revised version was published online in August 2006 with corrections to the Cover Date.     

In vitro pollen germination and tube growth of tomato (Lycopersicon esculentum Mill.) and its relation with plant growth

Summary In vitro pollen germination at 10°C, 14°C and 22°C of four groups of two pure line tomato varieties was compared with their plant growth at 19°C D/10°C N under controlled environmental conditions. Generally, pollen germination was slow at 10°C but after 6 h the percentage of germination was similar to that at 22°C. Maximum germination was obtained at 14°C already after 4 h. The longest pollen tubes occurred at 22°C. The two varieties within each group differed significantly from each other for percentage of pollen germination. For one group, this difference was greater at 10°C than at 22°C. The varieties in two groups also differed significantly for pollen tube growth. These differences in pollen tube growth rate were greater at 22°C than at 10°C. There was no clear relationship between pollen germination, pollen tube growth and plant growth in any of the four pairs of varieties. The results are discussed with regard to the possibility of pollen selection at low temperature in order to improve the efficiency of breeding for growth at low temperature.     

Genetic analysis of hexaploid wheat,Triticum aestivum using intervarietal chromosome substitution lines—protein content and grain weight

Summary The 21 intervarietal chromosome substitution lines of the cultivar Hope in Chinese Spring were used to analyse the genetic differences between the two cultivars Hope and Chinese Spring in grain protein content and grain weight.Only one chromosome of Hope, 5D, significantly influenced grain protein content of Chinese Spring. Its influence was of only minor effect and was to decrease protein content expression of Chinese Spring. It has been postulated that the genetic control of protein content, in this instance, is most likely due to many genes each of small effect.Five chromosomes of Hope influenced the 1000 grain weight value of normal Chinese Spring, all increasing its expression. Chromosomes 1A, 4A and 5B were of major effect and 3A and 6A of comparatively minor effect. A minimal estimate of five genes determines the difference in grain size between these cultivars. The possible evolutionary significance of the contribution of the A genome of bread wheat to grain size determination is discussed. On the basis of certain findings of this study, proposals are made for breeding for increased grain size in hexaploid wheat.     

Gamma-irradiation of cacao (Theobroma cacao L.) pollen: Effect on pollen grain viability,germination and mitosis and on fruit set

Summary Theobroma cacao pollen fertilization capability was studied after 0, 50, 100, 200, 500 and 1000 Gy gamma-irradiation. For all irradiation doses, no alteration of pollen grain viability and in vitro germination was observed. In situ, for all doses, pollen tubes penetrated into the styles and reached the ovules 20 hours after pollination. In vitro observations of the pollen grain nuclei after 20 hours incubation showed that pollen irradiation causes inhibition of the division of the generative nucleus. Fruit survival rate 30 days after pollination decreased as irradiation doses increased from 0 to 100 Gy, and over 100 Gy no fruit set was obtained. The possibility of using irradiated pollen as a method for obtaining haploid cacao plantlets is discussed.     

The progamic phase in Cichorium intybus L. Pollen tube growth in the style,incompatibility reaction and gametophytic competition

Summary Pollen germination and tube growth were studied following compatible, incompatible and pseudo-compatible pollinations in chicory. Pollen germination begins 3 minutes after compatible pollinations. The earliest pollen tubes reach the ovary 17 minutes later. Many of the later germinating pollen tubes are arrested and burst at the stigma papillae. In the transmitting tissue inhibitional effects due to negative interactions between pollen tubes are frequently observed. Complete self-incompatibility results in total inhibition of germination. In case of pseudo-self-compatibility, some pollen germinate but germination and stigma penetration are delayed and often result in pollen bursting. There is no self-incompatibility reaction in the transmitting tract but if the pollen tubes reaching this tissue are relatively numerous, negative interactions between them occur as after compatible pollinations. An hypothesis is presented which attributes the negative interactions between pollen tubes to the diffusion of a substance from the growing pollen tubes. This substance would also provoke pollen bursting on the stigma.     

The relation between wheat lines derived from the F2, F3, F4 and F5 generations for grain yield and harvest index

Summary The relationships between the F2, F3, F4 and F5 generations for grain yield were determined using random, pedigreed lines derived from each generation. The lines from two crosses were grown in plots at two sites over two years. In the first year, only F2 and F3 derived lines were available, but in the second year the F2 to F5 were grown.Correlations between lines in one generation and the mean of lines derived from them in a following generation increased as the generations were advanced. Correlations between consecutive generations were higher than those between generations two or three apart. Correlations between F2 and F5 derived lines, which indicate the effectiveness of selecting F2 lines, varied from 0.10 ns to o.49^** when lines from both generations were grown in the same environment. Correlations between years of lines from the same or different generations were low and often non-significant.Harvest index was measured on the F2 and F3 derived lines at the one site in the first year. Selection for improvement of grain yield using harvest index was no more effective than selection for yield directly, when considered across years.It is conclued that, while gains in yield can be achieved by selecting for yield in early generations, a foremost consideration needs to be the influence of different sites and years on the effectiveness of selection.     

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.     

Compatibility and incompatibility in witloof-chicory (Cichorium intybus L.). 1. The influence of temperature and plant age on pollen germination and seed production

Summary For the production of inbred lines and F₁ hybrids in witloof-chicory information is wanted on characteristics such as the incompatibility system. These characteristics can only be studied properly if the influence of temperature and physiological status of the plant on pollen germination and seed production is known. Investigations were carried out with 9 self-incompatible (SI) and 6 self-compatible (SC) clones in glasshouses of the IVT phytotron at constant temperatures of 10, 14, 17, 20, 23 and 26°C. In general, in vivo pollen germination percentages were rather low after self pollination with an optimum for germination around 17–20°C. No seeds were formed at the lowest temperature (10°C) while seed production for SC clones was usually (rather) good at higher temperatures. At 26°C seed production in some clones decreased. Both pollen germination and seed production decreased at the end of the flowering period. There was a rather positive relationship at e.g. 17 and 20°C between pollen germination after selfing and seed production. When no pollen germination was observed, no seed formation occurred. When pollen grains did germinate, seed development would not necessarily occur in all cases. So this relationship only enables negative mass selection for SC.     

The effect of temperature on seed set and embryo development in reciprocal crosses of wheat and barley

Summary The effect of temperature on seed set and embryo development in reciprocal crosses of barley and wheat was assessed in crosses involving two spring barley varieties (Betzes, Martonvásári 50) and one wheat variety (Chinese Spring). Detached tillers placed in nutrient solution were pollinated in controlled environments at constant day-night temperature regimes (12, 15, 18 and 21° C) with a light intensity of 30,000 lux and a relative humidity of 80%. When barley was used as the female, lower temperatures (12 and 15° C) produced the maximum seed set, whereas for the reciprocal cross, the highest temperature (21° C) produced the best seed set in the Chinese Spring × Betzes combination.Low temperature retarded the embryo development. The highest numbers of hybrid plants were produced at 18° C and 21° C in the barley × wheat cross and in the wheat × barley cross, respectively. Embryos of about 1.5 mm length in the barley × wheat cross, and of about 1.0 mm length in the wheat × barley cross germinated successfully. The smallest embryo giving rise to hybrid plants was 0.57 mm in the barley × wheat cross and 0.51 mm in the wheat × barley cross.     

The effect of the homoeologous group 5 chromosomes with different vrn loci on growth phases and quantitative characters of wheat

The differences between effects of homoeologous group 5 chromosomes on growth phases and agronomic characters were studied by using reciprocal substitution lines between a winter wheat cultivar with a high vernalization requirement (Mironovskaya 808) and one with lowvernalization requirement (Bezostaya 1), in which the presence of different recessive vrn alleles is supposed. The two cultivars and the substitution lines Mironovskaya 808 (Bezostaya 1 5A), Mironovskaya 808 (Bezostaya 1 5B), Mironovskaya 808 (Bezostaya 1 5D), Bezostaya1 (Mironovskaya 808 5A), Bezostaya 1 (Mironovskaya 808 5B),Bezostaya 1 (Mironovskaya 808 5D) were grown at 10 different sowing dates. The results showed that differences between the homoeologous group 5 chromosomes of Mironovskaya 808 and those of Bezostaya 1influenced the growth phases in addition to the impact by the genetic background and sowing date. We inferred from the analysis and comparison of their effect on vernalization response that vrn loci on these chromosomes influence growth phases. It is probably due to pleiotropic effects of the loci. The rare occurrence of significant interactions between group 5 chromosomes × sowing dates probably indicates independence of their effect. Agronomic characters were also markedly influenced by sowing date and the difference in backgrounds between Mironovskaya 808 and Bezostaya 1. A significant impact by at least two of the chromosomes on almost all studied characters was detected. The chromosomes affected the combined characters in the order5D>5B>5A and the positive value of the differences suggests that a content of Mironovskaya 808 chromosomes is more advantageous. It was possible to find certain indices in some agronomic traits, supporting the idea that the expression of some characters can also be connected to vernalization requirement and thus to the expression of the vrn loci. This supposition is most probable in the number of tillers and number of spikes. In some traits significant interactions occurred between homoeologous group 5 chromosomes × genetic background. Sporadic and low significance between homoeologous group 5 chromosomes × sowing dates suggest that the genetic effect of these chromosomes is independent of environmental conditions. This revised version was published online in July 2006 with corrections to the Cover Date.     

Interspecific hybridization in Cucumis L. II. The crossability of species,an investigation of in vivo pollen tube growth and seed set

Summary The crossability of 12 Cucumis species of African and Asiatic origin was studied in a diallel cross, in order to find ways to realise the cross between the common cucumber (C. sativus L.) and its wild relatives which carry resistances against diseases and pests.Self-pollinations and cross-pollinations within species gave normal pollen tube growth and seet set. The different accessions within a species, as a rule reacted alike in interspecific crosses. In crosses between African species different crossing patterns were found, viz. bilateral congruity, bilateral incongruity and unilateral incongruity. Within C. sativus all accessions intercrossed freely, except one, which displayed unilateral incongruity.Good seed was harvested from several crosses and in some cases embryo culture was needed for further development of seeds. No good seeds were obtained from any cross between a species of the African group and C. sativus L.     

Effects of homoeologous group 1 and 6 chromosomes of the Cappelle-Desprez (Bezostaya 1) substitution lines on aspects of bread-making quality of wheat

Summary The group 1 and 6 inter-varietal chromosome substitution lines of Cappelle-Desprez (Bezostaya 1) were intercrossed along with the donor and recipient varieties, Cappelle-Desprez and Bezostaya 1, to give 36 genetically different families. The analysis of the means of these families showed that variation in SDS-sedimentation volume fitted a predominantly additive model. There were no significant within or between chromosome interactions among the group 1 and 6 chromosomes. Nor was there any evidence for interactions between these chromosomes and those of the background. Significant dominance/within chromosome interactions amongst the background chromosomes were however detected. Some of the positive effects on SDS-sedimentation were associated with increased grain hardness. Chromosome effects on % grain protein were not correlated with SDS-sedimentation.     

Reaction of triticale,wheat and rye accessions to graminaceous Fusarium spp. infection at the seedling and adult plant growth stages

Summary Pathogenicity of 20 isolates of 12 Fusarium species recovered from triticale seed against seedlings of 14 varieties of winter cereals (triticale, wheat, and rye) was tested. The most pathogenic inoculum was a mixture of isolates (a composite isolate) of all the species. The following species were individually the most pathogenic: F. avenaceum, F. culmorum, F. sambucinum var. coeruleum, and F. graminearum. Winter triticale was more resistant to seedling blight than rye but more susceptible than wheat.Also reactions of 31 winter and 12 spring varieties of cereals to head inoculation with a composite isolate of 4 Fusarium spp. (F. avenaceum, F. culmorum, F. graminearum, and F. sambucinum var. coeruleum) was studied. In comparison to other cereals of similar type winter and spring wheat appeared to be the most susceptible while winter rye reaction was comparable to winter triticale. Spring and winter triticale varieties responded to head infection intermediately.There was no significant correlation between seedling and head reactions to infection with Fusarium spp. for winter rye and triticale. For winter wheat a negative trend was found. The above findings imply that screening of cereals at the seedling stage can not be used to predict the resistance to head blight. Nevertheless, resistance at the stage is highly desirable to prevent excessive damage of the crops due to the seedling blight incited by Fusarium spp..     

The inheritance of host plant effect on latency period of wheat leaf rust in spring wheat. I: Estimation of gene action and number of effective factors in F1, F2 and backcross generations

Summary Crosses were made between the highly susceptible Little Club and the partially resistant cultivars Westphal 12A, Akabozu and BH 1146 to obtain F1, F2 and backcross generations. Latency period (LP) was determined in plants inoculated at the young flag leaf stage with a monospore culture of race Flamingo of wheat leaf rust. Broad sense heritability of LP in the F2 averaged 0.8. The genes showed partial to almost complete recessive inheritance. Scaling tests indicated that additive gene action was the most important factor in the inheritance of partial resistance. The tests showed that there were no indications for additive x additive, additive x dominance or dominance x dominance interactions. The number of effective factors was estimated as one or two for Akabozu, three or more for Westphal 12A, and two or three for BH 1146. BH 1146 also possessed a (semi-)dominant gene for a lower infection type which was temperature sensitive in its expression. The genes of the various parents had unequal effect on LP.