Causal relationships between inbreeding, seed characteristics and plant performance in leek (Allium porrum L.)

Leek (Allium porrum L.) is an important outdoor vegetable in West Europe, where it is cultivated on about 30,000 ha. Most commercial cultivars of leek are open pollinated. One of the major problems with the crop is poor uniformity. Leek is an outbreeding species with up to 20% self-fertilisation. Much of the variation of open pollinated cultivars is explained by the strong sensitivity to in breeding depression after selfing. In this study we attempt to analyse in a systematic way the relationship between degree of selfing and agricultural performance in leek. The results obtained confirm a negative correlation between selfing and somea gricultural aspects important for vigour such as seedling emergence, plant growth and plant fresh yield. Also negative correlations between degree of selfing and seed individual weight and between selfing and seed individual size were found. The negative relationship between inbreeding and plant weight at harvest is further confirmed in an experiment in which the origin (cross-pollination orself-pollination) of each individual offspring plant analysed was determined using AFLP-markers. Finally, the correlation between chlorophyll deficiency genes and the loss of vigour that follows selfing was investigated. The results obtained demonstrate a significant decrease of pigment content in two generations of selfing. This revised version was published online in July 2006 with corrections to the Cover Date.     

The causes and consequences of meiotic irregularity in the leek (Allium ampeloprasum spp. porrum); implications for fertility, quality and uniformity

We have previously confirmed that Allium ampeloprasum spp. porrum, the leek, shows extensive and frequent quadrivalent formation during prophase I of meiosis, and that despite the resolution of most of these into bivalents some quadrivalents persist to metaphase I; in addition, a substantial number of univalents are found at this stage. In this paper we examine the later meiotic and post meiotic consequences of these irregularities in four leek cultivars and discuss their implications. Irregularities of chromosome segregation are observed at anaphase of both meiotic divisions, giving rise to micronuclei in dyad and tetrad stage pollen mother cells. It is shown that micronucleus frequency in tetrads is correlated with univalent frequency at metaphase I, suggesting that micronuclei originate largely from missegregation of univalents. Seedling populations of the same four cultivars were screened cytologically and shown to contain aneuploids at frequencies ranging between 4.3% and 8.4%. It is proposed that the majority of aneuploid seedlings originate from meiotic irregularity, especially univalent missegregation. It is very likely that the presence of this level of aneuploidy in the seedling populations contributes to the problems of phenotypic non-uniformity experienced by the leek crop, although its relative importance is difficult to assess. Further studies are required to establish the genetical and environmental contributions to meiotic irregularity and to assess the potential for selection for greater meiotic regularity in this material. This revised version was published online in July 2006 with corrections to the Cover Date.     

Breeding potential of Belgian landraces of leek ( Allium ampeloprasum L. var. porrum)

Western Europe is the world's greatest leek producing and consuming region. Belgium is situated in the centre of Western Europe and can be considered as a genepool of landraces of leek. Although the commercial varieties nowadays take over the successes of Belgian landraces, this paper deals with the breeding potential of the Belgian landraces. Typing the Belgian landraces was done in an observation trial together with 50 cultivars. Clustering analysis revealed that 4 of the Belgian landraces were autumn types, 2 landraces were of the early winter type and another 12 landraces were ranged with 6 commercial varieties to form the most winterhardy group of leek. For the important characteristics leaf colour and senescence resistance, the Belgian landraces were better than most of the commercial varieties. Although, for economically more important characteristics shaft length and some disease resistance they scored low. Finally, in this trial, the Belgian landraces were high yielding, especially in the late season. But also in the early season (harvest in September and November) some Belgian landraces were in the top 10 for yield. At harvest in February 9 of the 10 best yielding winterleeks were Belgian landraces. This proves that the Belgian landraces were well adapted to the soil and climate of this region. This revised version was published online in July 2006 with corrections to the Cover Date.     

Inbreeding and selection in swedes (Brassica napus ssp rapifera)

Summary The effect of inbreeding and selection within the old swede cultivar Scotia was examined. A decline in yield was shown by lines produced by selfing from the highest yielding progenies selected after open pollination. It is assumed that these progenies were from heterozygous plants. The yield of two groups of lines did not decline and from these, after a second generation of selfing, eight lines were selected with a mean dry matter yield 10 per cent higher than Scotia.Now part of Scottish Crop Research Institute, Dundee, Scotland.     

Plant regeneration via organogenesis and somatic embryogenesis in callus cultures derived from mature zygotic embryos of leek (Allium ampeloprasum L.)

Summary A high frequency plant regeneration system via organogenesis and somatic embryogenesis was established with callus cultures derived from mature zygotic embryos of different leek genotypes (Allium ampeloprasum L.). Four different callus types with varying morphogenetic potential were obtained. Relatively high concentrations of the auxin 2,4-dichlorophenoxy-acetic acid reduced callus weight and subsequent shoot regeneration and primordia formation of the callus. Shoot regeneration and primordia formation of the callus decreased after prolonged subculture on media containing 2,4-dichlorophenoxy acetic acid. A callus growth period of six weeks on Murashige and Skoog medium with 0.25–0.5 mg l^-1 2,4-dichlorophenoxy acetic acid showed the highest rate of shoot regeneration after transfer of callus to regeneration medium with 1 mg l^-1 kinetin.Differences between leek genotypes in callus type, callus weight, shoot regeneration and primordia formation were observed. Histological observations showed that plant regeneration took place, both via the pathway of somatic embryogenesis and organogenesis.Abbreviation 2,4-D 2,4-dichlorophenoxy acetic acid - MS Murashige and Skoog (1962) medium     

Resistance toPhytophthora porri in leek and some of its wild relatives

Summary Winter leek and related plant material was evaluated for resistance to white tip disease (Phytophthora porri) in the Netherlands. Significant differences in partial resistance were found between winter leek accessions in field tests with zoospore and natural inoculation. These differences were consistent over three years, and indicate that resistance can be improved through mass selection in winter leek cultivars. Some accessions of wild or cultivated relatives of winter leek with higher levels of resistance were found in a glasshouse test with zoospore inoculation. These sources of resistance were also highly resistant in field experiments over a two year period.     

The somatic karyotype of cabbage (Brassica oleracea ssp. capitata)

Summary The individual chromosomes of cabbage are identifiable in nucellar cells found in the ovaries of immature pistils by the combination of their relative length, arm ratios, secondary constrictions, chromatic and achromatic regions and the presence of satellites.     

Geographical distribution of allozyme patterns in shallot (Allium cepa var. ascalonicum Backer) and wakegi onion (A. × wakegi Araki)

Summary Shallot and wakegi were collected in the main islands of Indonesia, and in Japan, Korea, Taiwan, Malaysia, Thailand and Bangladesh. Five isozyme resolutions, phosphoglucomutase (PGM), glutamate oxaloacetate (GOT), glutamate dehydrogenase (GDH), esterase (EST) and peroxidase (POX) were employed for demonstrating inter-and intraspecific differences. A dendrogram separated 189 collected accessions into 25 types of wakegi onion and 18 types of shallot. All accessions of Japan, Korea and Taiwan were determined to be wakegi onion, whereas those of Bangladesh, Malaysia and Thailand were shallot. Twenty-six out of 165 Indonesian accessions indicated wakegi onion distribution in Sumatra, West Java province and in Sulawesi Island. This confirmed that there is mixed-cultivation of the two Allium species with no distinction made between them. Japan and Indonesia had respectively 12 and eight unique types of wakegi onion, while Korea had only one type. West Java showed the most various type of wakegi onion, whereas East Java had many types of shallot. Shallots collected from Bangladesh were distinetly different from those of South East Asian types.     

Female fertility of potato (Solanum tuberosum ssp. tuberosum) dihaploids

Summary Results of testcrosses and seed set data of the dihaploid breeding program at the Potato Research Institute in Gross Lüsewitz, Germany, indicate that the low level of female fertility could hinder success in crossing work. With the objective to reveal major components of female fertility, the behavior of 47 outstanding potato dihaploids (S. tuberosum ssp. tuberosum, 2n=2x=24) was assessed in testcrosses over a period of 3 years. Analysis of variance was carried out for the traits: number of seed per berry (S/B), number of intact seed per berry (IS/B), and number of non-germinable seed per berry (NGS/B). Significant sources of variation included cross combinations, years, and cross combination-year interactions. More than 20 S/B were obtained in 25.4% of all cross combinations performed over 3 years using up to three pollinators. Berry set was not correlated to seed set, but was in general determined by the pollinator used. The number of seedless berries (SLB/F) was related to neither total berry set nor seed production. The trait S/B remained stable over different years, and thus it deemed to be most useful as a measure of the level of female fertility in practical breeding work. Selection for female fertility in early generations of a breeding program is recommended to avoid limitations in the production of variability via sexual recombination.     

RAPD characterization of Brazilian barley (Hordeum vulgare ssp. vulgare) varieties

Thirteen varieties of Brazilian barley selected for malting qualities were analysed by RAPD (Random Amplified Polymorphic DNA). Amplification with 18 random primers generated 221 reproducible bands, of which 206 bands were polymorphic (93%):of this number, 137 fragments (62%) detected diversity among varieties and 56 bands (25.34%) allowed the distinction of varieties or groups of them. Variation was detected in all Brazilian varieties studied. Within-variety similarities estimated by Jaccard Similarity Coefficient ranged from 0.28 to 0.94, with averages ranging from 0.57 to 0.83, and an overall average of 0.72. Nevertheless, in the cluster analysis representatives of the same variety always fell into the same group and only later joined the other varieties. The average intervarietal similarities estimated by Jaccard Similarity Coefficient ranged from 0.45 to 0.62, with an overall average of 0.52. Many bands or combinations of bands which were responsible for the differentiation of all varieties were detected. Nevertheless, the majority of these bands cannot be considered as diagnostic markers because a great number of them did not occur in many representatives of the variety or had low intensity or even because they were not easily identified in the total pattern of bands.     

Antioxidant changes during postharvest processing and storage of leek (Allium ampeloprasum var. porrum)

The evaluation of the impact of postharvest processing and storage on the health benefits of vegetables is of great practical importance. Reports on their effect on the antioxidant capacity, polyphenol and S-alk(en)yl-l-cysteine sulfoxide (ACSO) content, i.e. isoalliin and methiin of the white shaft of leek (Allium ampeloprasum var. porrum), however, are limited. This study determined the levels of the antioxidant properties of leek from postharvest processing at the farm until refrigerated storage of 13 days at the consumer. Two cases were investigated, (1) leek sold as an entire plant and (2) leek with a large part of the green leaves removed, where the shafts are sold in a plastic package. The antioxidant capacity and the total content of phenolic compounds in the white shaft of the entire and packaged leek was stable during 13 days of refrigerated storage. A significant increase in the concentration of isoalliin was observed. Comparing the entire and processed/packaged leek, significant differences could be observed in antioxidant properties. The ACSO content in the white shaft of packaged leek was significant lower than the content in the white part of the entire leek. Therefore, the minimal processing step of cutting the green leaves and roots had an influence on the levels of antioxidant properties.     

Inbreeding depression in garden and glasshouse chrysanthemums: germination and survivorship

Summary Sixty-six chrysanthemum (Dendranthema grandiflora) inbred selections, noninbred cultivars and hybrids, and D. makinoi were the base populations from which up to three generations of inbreds were obtained using multiple plant descent. Each parent possessed pseudo-self compatibility (PSC), which allowed seed set following self pollination. Rapid-generation cycling (laboratory seed ripening to heart stage and subsequent embryo rescue) reduced generation time and minimized confounding maternal with zygotic inbreeding depression during post heart-stage seed development. Selection criteria were male/female fertility and PSC. Two stages of the life cycle were chosen to evaluate inbreeding depression: germination (seed development to germination) and survivorship (fertile individuals at anthesis). PSC was environmentally interactive and genotype dependent, causing variable levels of self seed set between and within populations and generations. By the end of the second generation, families from all noninbred cultivars, D. makinoi, and one F₁ hybrid were eliminated from the experiment due to self incompatibility and/or inbreeding depression. Post-germination inbreeding depression was severe in several advanced inbreds. Inbred progeny of most F₁ or F₂ hybrids expressed less or equal amounts of inbreeding depression compared to advanced inbreds. Linear regression coefficients for either germination or survivorship on percent homozygosity were negative. Correlation coefficients between percent germination and survivorship (as a percent of seed set) were highly significant for Minnesota inbreds (r=0.67, P0.002) and hybrids (r=0.67, P0.006). The correlation coefficient was higher when percent germination and survivorship (as a percent of germinated seedlings) were compared (r=0.95, P0.001).Abbreviations F Fisher's Coefficient of Inbreeding - IBD Inbreeding Depression - PSC Pseudo-self Compatibility - RGC Rapid Generation Cycle - SI Self Incompatibility     

Breeding for resistance to neck rot (Botrytis allii) in onions (Allium cepa)

Summary For 10 years the method developed by Van der Meer et al, (1970) to test neck rot resistance in onions has been applied in a breeding programme. From experiments with market varieties, inbred lines and half-sib families it is concluded that neck rot resistance as determined by this method is a quantitative character, which shows continuous variation and is in part fixable by selection. The practical importance of this type of neck rot resistance is not yet clear.Report no. 68     

Heterotic patterns in hybrids involving cultivar-groups of summer squash, Cucurbita pepo L.

Heterotic response for total yield among five cultivar-groups of summer squash, Cocozelle, Vegetable Marrow, Zucchini, Crookneck and Straightneck, was studied in two sets of diallel crosses evaluated in two harvest seasons. Each diallel cross included two representatives of each the five cultivar-groups. Griffing and diallel GGE biplot (genotype main effect and interaction) analyses were carried out. The relative importance of the specific combining ability variance on total variance was greater than that reported earlier for intragroup crosses. Highly significant heterosis was manifested only in combinations of Crooknecks or Straightnecks crossed to Cocozelles, Vegetable Marrows or Zucchinis. GGE biplots showed a consistent arrangement where testers of ssp. texana interacted with cultivars of ssp. pepo and vice versa. We propose that the heterotic pattern of texana-pepo crosses could be potentially used to achieve increased yields in hybrids by introgressing one group into another in a manner that would maintain fruit attributes of the cultivar-group. This revised version was published online in July 2006 with corrections to the Cover Date.     

Genetic evaluation of cultivated garlic germplasm (Allium sativum L. and A. ampeloprasum L.)

Garlic cultivated in rural farms of South Italy is often a heterogeneous clone population, which can comprise different cytotypes. A collection of cultivated garlic from the University of Basilicata gene-bank, was evaluated for ploidy level, and 16 different morphological traits. Out of50 accessions, 7 were hexaploids (Allium ampeloprasum L.) and43 diploids (Allium sativum L.). Significant differences in yield were observed within and between ploidy levels. The heritable traits most correlated with yield were plant height (h² = 0.62) within diploid types and neck diameter (h² = 0.75) within hexaploids. Discriminant analysis did show that four characters (leaf basal width, total n^° of leaves, clove diameter and neck height) were able to correctly discriminate all germ plasm accessions between the two species. This revised version was published online in August 2006 with corrections to the Cover Date.     

Chromosome and cytoplasm analyses of somatic hybrids between onion (Allium cepa L.) and garlic (A. sativum L.)

Chromosomes and cytoplasms were analyzed in two lines of a somatic hybrid between onion (Allium cepa L.) and garlic (A. sativum L.). One line of the somatic hybrid had 40 chromosomes and the other 41chromosomes. Genomic in situhybridization successfully revealed the chromosome constitution of the two lines. One line had 20 chromosomes from onion and17 chromosomes from garlic, and the other had 21 chromosomes from onion and 17chromosomes from garlic. Interestingly, both lines had three chimeric chromosomes. PCR-RFLP analyses of chloroplast and mitochondrial DNAs of both lines showed that these were identical to the onion parent. This revised version was published online in July 2006 with corrections to the Cover Date.     

Transfer of resistance to race 2 of Plasmodiophora brassicae from Brassica napus to cabbage (B. oleracea ssp. capitata). III. First backcross and F2 progenies from interspecific hybrids between B. napus and B. oleracea ssp. capitata

Summary The first backcross and F₂ progenies from triploid F₁ and tetraploid F₁ hybrids between B. napus and 2x and 4x B. oleracea ssp. capitata (cabbage) were studied for their general morphology, resistance to race 2 of the clubroot pathogen, chromosome number and meiotic chromosome behavior. No linkage was apparent between resistance and the major morphological characters. Unreduced gametes played a large part in the successful formation of seed of the B₁ and F₂ progeny. B₁ plants with low chromosome numbers were selected for use in recurrent backcrosses. The potential use of anther culture to extract gametic progenies from resistant B₁ and F₂ plants with higher chromosome numbers was suggested. The presence of homoeologous pairing observed in all the plants is considered advantageous for selecting suitable progeny in later generations.     

Use of the gene pools of Triticum turgidum ssp. dicoccoides and Aegilops squarrosa for the breeding of common wheat (T. aestivum), through chromosome-doubled hybrids

Summary Triticum turgidum ssp. dicoccoides (wild emmer wheat, AABB, 2n=28) and Aegilops squarrosa (goat grass, DD, 2n=14) comprise a rich reservoir of valuable genetic material, which could be useful for the breeding of common wheat (T. aestivum, AABBDD, 2n=42). Many accessions of both wild species, most of them selected for resistance to stripe rust, were used to make amphiploids. Two strategies were applied: (1) the production of autopolyploid cytotypes of the wild species, followed by hybridisation, and (2) the production of allotriploid interspecific hybrids, followed by doubling of the number of chromosomes. The first route was unsuccessful because of failure of the crosses between the autopolyploid cytotypes, possibly due to incongruity between the two species and to reduced fertility in the autopolyploid cytotypes. The second route yielded the desired synthetic hexaploids. However, the rate of success of the crosses was low and there were great differences between years, and within years between crosses. Embryo rescue was applied to obtain the primary hybrids (2n=21), which were highly sterile and had on average 0.3 bivalents and 20.4 univalents per pollen mother cell. Various abnormalities were recorded. Doubling of the number of chromosomes sometimes occurred spontaneously or was brought about by colchicine treatment. The large scale of the interspecific hybridisation programme ensured that one-third of the female and one-sixth of the male accessions were represented in the synthetic hexaploids.     

Self-pollination in swedes (Brassica napus ssp Rapifera) and its implications for cultivar production

Summary The degree of selfing on open pollination was examined in four swede cultivars. Three cultivars had over 80 per cent selfing, whilst the fourth had less than 55 per cent. The inbreeding coefficient with these levels of selfing, assuming the cultivars were at equilibrium, would be 0.69 and 0.37 respectively. The cultivar with the high level of outcrossing was found to possess a degree of self-incompatibility. The structure of such populations is demonstrated, and the implications for cultivar production are discussed.     

Somatic hybrids between Gossypium hirsutum L. (4×) and G. davidsonii Kellog (2×) produced by protoplast fusion

Somatic hybrids were obtained from electrofused protoplasts derived from embryogenic suspension cultures of tetraploid cotton (G. hirsutum L. cv. Coker 201) and embryogenic callus of diploid wild cotton G. davidsonii. The regenerants were initially identified as hybrids by RAPD (random amplified polymorphic DNA) analysis. Subsequently, observation on chromosome counting, morphology and SSR (simple sequence repeat) confirmed the hybrid status. Cytological investigation of the metaphase root-tip cells of the regenerated plants revealed there were 74 to 84 chromosomes in the plants, close to the expected 78 chromosomes. SSR analysis revealed the regenerated plants contained specific genomic fragments from both fusion partners, further confirmed their hybridity. The morphology of the plants was intermediate between the two fusion partners. The regenerants were difficult to develop into mature plants because their roots browned and they wilted from the stem apex before forming 3 to 5 true leaves. The hybrid plants were transferred to soil by grafting in vitro onto rootstocks.