Breeding tomato for pollen tolerance to low temperatures by gametophytic selection

Haploid selection for traits related to pollen cold tolerance in tomato was performed in segregating populations derived from a Lycopersicon esculentum × L. pennellii hybrid. BC₁ populations were obtained by combining normal and low temperature treatments on two stages of pollen development: pollen formation, and germination and pollen tube growth. F₁ hybrids were cultivated under low and normal temperatures and their pollen was used to pollinate L. esculentum plants at low and normal temperatures. The four BC₁ populations obtained were tested for the quality and quantity of pollen produced at low temperatures. The population obtained by cold treatment at both stages had a significantly improved pollen germination ability at low temperatures. The two other coldselected BC₁ populations showed no differences compared with the unselected BC population. A second cycle of pollen selection, corresponding to BC₂, was applied in order to test its persistence in the subsequent generations and the possibility to further improve the character. This second cycle showed no improvement although some plants retained the high pollen germination ability at low temperatures that was observed in the first cycle. Hence, gametophytic selection of some characters related with tomato pollen performance may be feasible, at least for the first cycle of selection.     

Selection for Oil Content in East Indian Lemongrass

The response of oil content to phenotypic selection was studied in a population of lemongrass raised from seeds of a clonally propagated variety SD-68. During November-December 1982, 400 plants were evaluated for oil content. Five per cent of the evaluated plants were selected for high and low oil content and were separately intermated in isolated polycross (PC) blocks in November 1983-February 1984. The populations derived from high and low oil PC blocks were evaluated, along with the unselected parental population and OD-19, a variety used as a local check, in a randomized complete block design, during October 1984, January and May 1985 for leaf yield, oil content and citral content in oil. The high oil PC population had a significantly higher oil content than the unselected parental population and the local check OD-19, during all the three seasons. The low oil PC population had a significantly lower oil content than the unselected parental population in January and May 1985. Over seasons, the high oil PC population had 0.15 per cent higher oil content than the unselected parent population. Realized heritabilities for high and low oil content were 0.52 and 0.50, respectively. No significant differences were observed between the different populations for leaf yield in all the three seasons and for citral content in October 1984. In January and May 1985, the differences in citral content were not related to the differences in oil content.     

The production of aneudihaploids in Solanum tuberosum L. group Tuberosum (the common potato)

Summary Trisomic cytotypes were produced in dihaploid (diploid) plants of Solanum tuberosum L. Group Tuberosum, the common potato, according to two methods. Firstly, the aberrant types were selected, through chromosome counting, from parthenogenic dihaploid offspring of tetraploid-diploid crosses. In dihaploid populations from twelve tetraploid potato varieties the frequencies of aneuploids ranged from 3.5 to 11.0%. About 95% of these aneuploids had only one, and the others not more than two extra chromosomes. Secondly, the aneuploids were produced from triploid-diploid crosses. Seedset strongly depended on the crossability of the parental plant material, and the best results were obtained when the motherplants were grafted onto tomato. On avarage the three most successful cross combinations resulted in approximately 0.7 berries per pollination and 6 seeds per berry. With regard to seedsize the seed could be divided in two groups, viz, normal and small seeds. Half of the seed did not germinate or produced inviable seedlings, especially among the small seed group. About 93% of the plants was neuploid, with 25, 26 and 27 being the predominant chromosome number. It was concluded that the production of trisomics was the most successful through triploid-diploid crosses. The results were discussed with reference to the chromosomal behaviour in the meiosis in triploid plants (Lange & Wagenvoort, 1973a). It thus was possible to relate the low seedset to the distribution of chromosomes in the meiotic anaphases. On the one side this distribution resulted in a limitation of the avalability of gametes with monohaploid and near-monohaploid chromosome numbers, while on the other side the macrospores with higher chromosome numbers seemed to be inviable. Finally a crossing scheme was presented for transmitting the trisomic condition into a genetic background with better homogeneity and more homozygosity.Extended version of a lecture presented at a Eucapia meeting about Potato Dihaploids, held at Wageningen (Netherlands). See Lange and Wagenvoort, 1973b.     

Evaluation of Self and Polycross Progeny Testing in Birdsfoot Trefoil (Lotus corniculatus L.) for Forage and Seed Yield

Self and cross fertility, forage and seed yield of parents, self (Sl), and polycross (PX) progenies were measured in a selected and an unselected population of birdsfoot trefoil (Lotus corniculatus L.), Self fertility was significantly less than cross fertility in both populations. In a field planting S1 progenies had lower forage yield in 1979 and 1980 and seed yield (relative to the parents) was less in the selected population than the unselected in 1979. All correlation coefficients between years and progenies were non-significant for forage yield bur the parents-poly cross progeny correlation was significant for seed yield in the selected population. Percentage survival of plants after cutting in 1980 was higher in the selected population, and the parents and PX progenies survived better than S1 progenies in both populations. The better performance of the selected population may be due, in pan, to higher frequency of favorable genes resulting from four cycles of recurrent selection, S1 progeny testing does not appear to be worth while in birdsfoot trefoil because of low self fertility and the reduced vigour of the progeny.     

Effect of plant density on selection for seed yield in two population types of Phaseolus vulgaris L.

Summary Pedigree selection for seed yield, using early generation yield tests, was practiced from the F₂ to F₇ in two populations of common bean (Phaseolus vulgaris L.) in three plant densities: 66, 133, and 266 thousand plants ha^-1 at CIAT-Palmira, Colombia. The six highest yielding lines selected from each plant-density, along with 13 parents, were evaluated in a 7×7 lattice design with three replications at the three densities utilized for selection, and at 399,000 plants ha^-1 in 1988 and 1989.Based on the mean performance of selected lines and the mean of the parents, selection for seed yield was effective in all densities in both populations. However, none of the lines selected from the population within the race Mesoamerica (TC 4673) significantly outyielded their best parent under any plant density. The highest yielding lines selected from the interracial population (TR 4635) outyielded their best parent irrespective of the plant-density used for selection. The highest yielding line originated from the highest density used for selection. Low density was neither good for selection nor for evaluation and identification of high-yielding cultivars of common bean. There was no significant difference between the mean yield of lines selected at the intermediate and high population densities. The effects of plant density, year, and their interactions were significant for seed yield.     

The production of yellow-seeded Brassica napus (AACC) through crossing interspecific hybrids of B. campestris (AA) and B. carinata (BBCC) with B. napus

To transfer the genes for yellow seed coat from both genomes A and C to B. napus (AACC), the hexaploid of Brassica (AABBCC) was synthesised from reciprocal interspecific crosses between yellow-seeded B.campestris (AA) and B.carinata (BBCC). The hexaploid with 27 pairs of chromosomes was red-seeded which showed that genic interaction existed in the trigenomic plants for the colour of the seed coat. Hundreds of hybrid seeds were obtained from crosses between the red-seeded hexaploid and partial yellow or brown-seeded varieties of B. napus as pollen donor. The majority of the hybrid plants (AABCC) were self fertile with brown seeds. It appeared that the chromosomes of the B genome were excluded during the meiosis of the pentaploid and a high proportion of the genetically balanced AC gametes could be produced. The fertility of the F2 population was increased and even reached normal levels for some plants. Seventy-three plants with the yellow-seeded character were isolated from 2590 open-pollinated F2 plants, most with increased fertility. After two successive self-pollinations, 18 lines produced yellow seeds and no brown seeds segregated from these populations. The morphology of the novel yellow-seeded plants was basically towards B. napus. Esterase isoenzyme electrophoresis showed that the plants contained some of the genetic background of B. campestris, B. carinata and B. napus. Cytological analysis has shown that at least some yellow-seeded lines have the B.napus AACC genome composition with 38 chromosomes and normal meiotic pairing. This revised version was published online in July 2006 with corrections to the Cover Date.     

Further results on chromosome stability of hexaploid Triticale

Summary Chromosome variation was studied in bulk populations of 27 advanced strains of hexaploid Triticale. Average frequency of euploids (2n=42) in the 27 strains analyzed was 81.5% with a range from 57.2% to 100.0%. Three strains showed particularly high aneuploid frequency in the bulk population; the average frequency for the three strains was 39.0%, while the mean for the other 24 strains was 14.8%. The progenies of 42-chromosome plants derived from the aforementioned three strains had 89.4% euploids on an average. It is assumed that a monosomic plant was selected in a single-plant (or spike) selection during the breeding period. The frequency of aneuploids is very high, even after several generations of breeding. Routine chromosome checking is recommended in breeding Triticale and other cytologically unstable induced amphiploids.Contribution No. 175 of the Department of Plant Science, University of Manitoba, Winnipeg, Canada.     

The stabilization of chromosome numbers and the maintenance of euploidy in Brassicoraphanus

Summary To examine whether chromosome numbers of Brassicoraphanus (amphidiploids between Brassica japonica Sieb. and Raphanus sativus L.) are stable or not, the following four items were studied with some plants during the 2nd–11th generations: (1) chromosome numbers of open-pollinated progenies from eu-, hyper-, and hypoploids, (2) chromosome distribution at metaphase II in these plants, (3) frequency of euploids in relation to flower colour and generation, (4) seed fertility in eu-and aneuploids in relation to flower colour. In early generations, hyper-and hypoploids were frequently produced from euploids. In later generations, however, the chromosome number became less unstable. In euploids (2n=38), chromosome numbers at metaphase II showed some variation, and the mean frequency of the euploid chromosome number (n=19) was approximately 78%. This value was almost the same in white-and yellow-flowered plants through early and late generations. Nevertheless, yellow-flowered plants tended to produce euploids more frequently than did white-flowered ones. It is assumed that the difference in euploid productivity between yellow-and white-flowered plants is due to the difference in seed fertility between them. The progeny of each hypoploid showed higher chromosome numbers than their parents. The progeny of each hyperploid showed lower chromosome numbers than their parents: they were approaching to euploidy. This phenomenon, together with higher fertility of euploids and lower fertility of aneuploids, will favor the maintenance of euploidy of this strain.     

Occurrence of 2n and jumbo pollen in the Brazilian ginseng (Pfaffia glomerata and P. tuberosa)

An analysis of microsporogenesis was carried out in four populations of Pfaffia tuberosa (2n=4x=68) and in ten populations of P. glomerata(2n=2x=34). Monads, dyads and triads were reported at high frequency in all populations of P. tuberosa. In P. glomerata, on the other hand, only four populations presented monads, and at low frequency. Dyads and triads were observed in both species, albeit at a higher frequency in P. tuberosa populations. Certain abnormalities of spindles in the second division, such as sequential and tripolar spindles, led to triad formation by the rejoining of two chromosome sets. The precise causes of monad and dyad formation have not been found during meiosis, although evidence exists that dyads originated by omission of second meiosis and monads by omission of both meiotic divisions. This revised version was published online in July 2006 with corrections to the Cover Date.     

Stabilization of new types of diploids (2n=22, 24) through selfing of aneuploids (2n=21, 22) derived from crossing of sesquidiploids (2n=29, AAC) and Brassica campestris (2n=20 AA)

Summary Aneuploids with 2n=21 and 2n=22 derived from crossing of sesquidiploids (2n=29, AAC) and Brassica campestris (2n=20, AA) were selfed successively in order to follow the changes in chromosome number of the progenies for three consecutive generations. Progenies with 2n=22, 23 and 24 obtained after selfing of S₀ generation and the succeeding S₁, S₂ and S₃ generations were analyzed in terms of pollen stainability, % seed set as well as cytogenetically based on meiotic behaviour with the aim of determining the possibility of addition of one or more alien chromosomes into n=10 species which may lead to differentiation of single or plural disomic addition lines. The generation of aneuploids with 2n=21 progressed in such a way that most plants seem to revert to the 2n=20 chromosome number of B. campestris after selfing. From 2n=22 aneuploids, however, the succeeding progenies showed high frequency of plants with two additional chromosomes which accounted for 50.6% and 52.9% of total S₃ progenies via 2n=22 and 2n=24 S₂ generations, respectively. The meiotic behaviour of these progenies indicated evidence for a rule governing the frequency distribution of chromosome number among these addition lines and high possibility to breed such disomic plants with 2n=22. A method of selecting stable aneuploids was suggested in addition to the possible role of pollination biology at various processes of such breeding program.     

Cytological studies of 2n male gamete formation in sugarcane, Saccharum L.

The germplasm for modern sugarcane cultivars (Saccharum spp. hybrids)has been derived principally from S. officinarum (2n = 80), and S. spontaneum (2n = 40 to 128). Diploid gamete formation has been significant in developing cultivated sugarcane, but the cytological basis for the processes involved is not clearly understood. This research investigated microsporogenesis in nine clones of Saccharum spp. Hybrids and in S. officinarum and S. spontaneum. Diploid gamete formation occurred in all 11 lines, but was least frequent in S. spontaneum and S. officinarum which produced 0.5% and 0.8%2n gametes, respectively. In the hybrid lines, 2n gametes were formed infrequencies ranging from 0.9% to 4.4%. Cytological evidence was obtained for dyad and triad formation during microsporogenesis. Detailed analysis of chromosome behaviour at meiosis indicated that 2n male gamete formation is probably attributable to the absence of cytokinesis rather than a combination of asynchrony and non-disjunction. The clones were ranked on the basis of the frequencies with which they formed 4 × 1n microspores and the data were analysed using χ ² tests for homogeneity. These established that theSaccharum spp. hybrids could be designated as either ‘high’ or ‘low’ frequency haploid gamete producers. Conversely, the latter group, which formed diploid gametes most frequently (2.2%–4.4%), can be described as high frequency diploid gamete producers. The identification of clones most frequently forming diploid gametes may facilitate the more rapid recovery of desirable sugarcane genotypes because such clones could be selected for preferential use in clonal improvement. This revised version was published online in July 2006 with corrections to the Cover Date.     

Response to divergent selection for ease of particle size reduction of dried leaves of smooth bromegrass ( Bromus inermis Leyss) and correlated effects on nutritive value indicators and plant fitness

The rate at which forage can be broken down and cleared from the rumen is a major constraint on voluntary intake by ruminants. Responsiveness to selection has been demonstrated for physical characters possibly related to the ease of breakdown of forage grasses and is a prerequisite for the exploitation of these characters in a breeding program. The objective of this study was to evaluate at two locations the response to one cycle of divergent selection for the ease of breakdown, or particle size reduction index (PSRI), of dried leaf blades in four populations of smooth bromegrass (Bromus inermis Leyss.) grown as spaced plants and the correlated response in neutral detergent fibre (NDF), in vitro dry matter digestibility (IVDMD) and on seedling performance and second year spread as indicators of plant fitness. Divergent selection for PSRI resulted in significant differences between high and low groups in all populations. The contrast for asymmetry of response was not significant, but response tended to be higher in the downwards direction relative to an unselected group. Realised heritabilities ranged from 10–21% when the unselected group was used to represent the parental generation. In the most responsive population, WB19e, seedling growth was poorer at both sites and seedling mortality higher at one site in plants selected for increased PSRI. Selection for PSRI had no effect on IVDMD but there were closely correlated changes in NDF (r = –0.73 across populations, p<0.01). However, comparisons between populations suggested that differences in PSRI independent of NDF also existed. PSRI can be exploited in a breeding program if gains in forage intake by ruminants, independent of changes in NDF, are demonstrated. This revised version was published online in July 2006 with corrections to the Cover Date.     

Increasing 2n gamete production in diploid alfalfa by cycles of phenotypic recurrent selection

Two cycles of selection (3rd and 4th) were carried out in a diploid population of alfalfa to improve the frequency of 2n gametes. All the plants produced unreduced gametes, except one in the 2n egg population. The average seed set, used as a measure of 2n gametes production, increased significantly in the 2n pollen selection. Conversely, the increase was not significant in the case of 2n eggs. The improvement in 2n pollen production was confirmed cytologically. These results, like those of the first two cycles of selection, suggest that 2n pollen production is controlled by few major genes while 2n egg production is controlled by one recessive major gene and influenced by minor genes. The mutants selected have been utilized for producing heterotic tetraploid progenies through Bilateral Sexual Polyploidization (BSP).     

Genetic variation in cocksfoot (Dactylis glomerata L.) populations for mottle virus resistance

Summary Highly significant and continuous variation in the degree of injury due to cocksfoot mottle virus (CfMV) infection was found among clones of 7 cocksfoot (Dactylis glomerata L.) populations, taken at random among surviving plants in 9 year old swards. Broad sense heritabilities for CfMV injury ranged from 0.52 to 0.91 within populations, and genotypic correlations between CfMV injury and total DM-yield ranged from –0.46 to –0.88. The relationship between CfMV injury and DM-yield could be described by a second degree curve. CfMV injury was to some extent genotypically correlated in the positive direction with crude protein content and digestibility, and negatively with crude fiber content, especially in the first cut. Artificial infection of plants from the original populations with CfMV in a simulated sward experiment, did not reveal any significant differences between populations in tolerance towards CfMV infection. Infection caused a 54.5% increase in the number of dead plants per plot, relative to uninfected plots, but only a 4.5% reduction in dry matter yield. There is a clear tendency that synthetic populations selected for high yield are less tolerant towards CfMV infection than unselected or low yielding populations. The resistance found in these cocksfoot populations is a sort of tolerance, and the results indicate that the level of tolerance can be increased by selection.     

Screening <Emphasis Type="Italic">A. ventricosa</Emphasis> populations for 2n gametes

Sexual polyploidization via the formation of 2n gametes has been acknowledged as the most significant evolutionary mode of polyploidization among plant species. The present study was conducted in order to determine whether 2n gametes are present in the C-genome diploid Avena ventricosa Bal. ex Coss., a species that contributed to the evolution of the cultivated hexaploid species (Avena sativa L). Individual plants belonging to four different Cypriot populations, were screened for pollen grain size variation with the aim to distinguish 2n gametes. Avena ventricosa ARI00-845 was identified to produce large pollen grains at a low percentage (1.21%). Subsequent analysis using flow cytometry confirmed the presence of 2n gametes in the pollen. Cytogenetic analyses of pollen mother cells revealed cells with twice the typical chromosome number at metaphase I (i.e., 28 chromosomes). We postulate that irregularities in cell wall formation preceding meiosis could have contributed to the mode of chromosome doubling.     

Investigations on the breeding behaviour of aneuploids of guava

Summary The breeding behaviour of aneuploids of guava (Psidium guajava L.) such as trisomic, tetrasomic, and higher aneuploids has been studied. Reciprocal crosses between aneuploids and diploids indicated less than 100% crossability. The aneuploids when used as male parents crossed less frequently than as female parents and certain aneuploids crossed more readily than others. Differences were observed in fruit size, fruit weight, and seed number in the reciprocal crosses.The extra chromosome was found to be transmitted through both the egg cell and the pollen. However, the frequency of transmission was greater through the egg cell than the pollen. As high as 26% transmission of extra chromosomes were obtained through the egg cell. There was no clear cut difference between trisomics and higher aneuploids with regard to the frequency of transmission of extra chromosomes.     

Occurrence of numerically unreduced (2n) gametes in Alstroemeria interspecific hybrids and their significance for sexual polyploidisation

The F1 hybrids of seven diploid Alstroemeria species (2n=2x=16) were investigated for the production of numerically unreduced (2n) gametes and their mode of origin. Based on a survey of 17 interspecific hybrid combinations,consisting of 119 genotypes, it was found that the F1 hybrids of Chilean-Brazilian species mostly produced first division restitution (FDR) 2n gametes. These F1 hybrids were self-pollinated in order to obtain F2 seeds, which was an indication that the F1 plants also produced 2neggs simultaneously. All the F2 progeny plants were typical allotetraploids, most of which formed 16 bivalents and a small proportion formed multivalents during metaphase I stages of meiosis. Through genomic in situ hybridisation (GISH) it was proved that multivalent formation in F2plants, derived from A. inodora ×A. pelegrina hybrid, was due to homoeologous recombination but not from reciprocal translocations. In order to test the segregation pattern of the recombinant chromosomes, an F3 population from one genotype, P6C49-6, was investigated. The recombinant chromosomes assorted independently from each other supporting the hypothesis that the segregation of chromosomes in ring quadrivalents did not behave like those in translocation heterozygotes. It was concluded that in allopolyploids of Alstroemeria,bilateral sexual polyploidisation could accomplish genetic recombination by both homoeologous crossing-over as well as through the assortment of chromosomes. This revised version was published online in July 2006 with corrections to the Cover Date.     

Comparative efficiency of honeycomb and conventional pedigree methods of selection for yield and fiber quality in cotton (Gossypium spp.)

The effectiveness of honeycomb pedigree selection (HPS) as compared to conventional pedigree selection (CPS) was studied in one intraspecific (G. hirsutum L. × G. hirsutum L.) cross population (population I) and one interspecific (G. hirsutum L. × G. barbadense L.) cross population (population II). Combined selection for yield and lint quality traits was applied for four cycles at two locations for population I and atone location for population II. Finally, the best F₆ lines derived by each method, together with the unselected population derived by single seed descent (SSD) and three check cultivars, were tested in comparative experiments, separately for each cross, at the same locations. In both populations the analysis of variance indicated significant differences among the groups of the material tested for seed cotton yield, mean boll weight, micronaire reading, staple length, and uniformity ratio. No significant differences were found with respect to plant height, lint percentage, and fiber strength in population I and with respect to lint percentage and fiber strength in population II. In population I, on the basis of mean seed cottonyield and number of superior lines derived by each method as compared to the check varieties and the unselected SSD population, HPS-lines were superior to CPS-lines for seedcotton yield, fiber length and boll weight. HPS-lines, however, did not differ significantly in seedcotton yield from the best check cultivar Eva. Finally, the material derived by CPS was earlier in maturity than the material derived by HPS and SSD. Also in population II, on the basis of mean seedcotton yield and number of superior lines derived by each method as compared to the unselected SSD population, HPS-lines were superior to CPS-lines. No significant differences, however, were identified between the material of the two methods for lint quality traits. It was concluded that in both populations HPS was more effective than CPS in identifying lines with high yielding ability and good lint quality. This superiority of HPS is attributed, at least partially, to its effectiveness in early generation selection. This revised version was published online in August 2006 with corrections to the Cover Date.     

Trifolium ambiguum as a source of variation in the breeding of white clover

Summary Hybrids between T. ambiguum and T. repens were successfully established after ovule culture. The hybrids were male sterile but one of the hybrids produced a single seed when backcrossed to T. repens, while the other produced a significantly greater number of seeds. The majority of the backcross hybrids from the latter had 48 chromosomes, being produced from the fertilisation of unreduced gametes of the F1 hybrid by T. repens pollen. Regular meiosis in the BC1 hybrid resulted in the production of balanced gametes containing the full complement of T. repens chromosomes and the polyhaploid set of T. ambiguum chromosomes. There was no difficulty in establishing a substantial population of BC2 hybrids. The meiotic behaviour of the BC1 hybrid indicated that it should be possible to transfer genes from T. ambiguum into white clover T. repens. The significance of the results in terms of the use of alien variation in the improvement of white clover is discussed.     

Genome Size Variation in Maize Populations Selected for Cold Tolerance

Previous studies have indicated a relationship between genome size and cold tolerance in plants. Many species adapted to growth in cool environments have large genome sizes. These studies are based on interspecific DNA content variation. In this study, the nuclear DNA content of eight maize populations was determined. These populations were obtained from the University of Nebraska and represent populations selected for cold tolerance and their respective unselected original populations. Intraspecific DNA content variation was observed between the selected and unselected populations. Upon assessing the data based solely on cold tolerance, no clear relationship between genome size and cold tolerance was apparent. When both freeze tolerance and cold tolerance were considered, populations which were cold tolerant and exhibited a certain degree of freeze tolerance were observed to have significantly larger genomes relative to the unselected populations. Thus, it appears that the relationship between intraspecific genome size variation and cold tolerance is similar to the relationship between interspecific genome size variation and growth at cooler temperatures in plants.