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  • 1
    Electronic Resource
    Electronic Resource
    Springer
    Plant cell reports 18 (1998), S. 229-234 
    ISSN: 1432-203X
    Keywords: Key words Citrus ; Washington navel orange ; Agrobacterium tumefaciens ; Genetic transformation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Transgenic Washington navel orange [Citrus sinensis (L.) Osbeck] plants were obtained using Agrobacterium-mediated transformation of seedling epicotyl tissue. An average of 45% (58 out of 128 segments) of the epicotyl segments produced shoots expressing the β-glucuronidase (GUS)-intron reporter gene when using Agrobacterium strain C58 C1, compared to 29% (38 out of 128 segments) for EHA101-5 and 0% for LBA4404. Co-culture of 21-day-old Washington navel epicotyl stem segments gave greater transformation efficiency than co-culture of 35- or 56-day-old stem segments. After 6 weeks, regenerated shoots were micro-grafted in vivo onto seedling rootstocks of Carrizo citrange. Stable integration of the transgene sequence was confirmed by expression of the plant intron-containing GUS gene, PCR and Southern hybridization. The apomictic (non-zygotic) state of the transgenic plants was confirmed by isoenzyme and random amplified polymorphic DNA analyses. More than 50 transgenic plants have been obtained and are growing in the greenhouse.
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  • 2
    Electronic Resource
    Electronic Resource
    Springer
    Theoretical and applied genetics 74 (1987), S. 162-167 
    ISSN: 1432-2242
    Keywords: Gossypium hirsutum L. ; Verticillium dahliae Kleb. ; Generation means analysis ; Minimum chisquare ; Genetic segregation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Three crosses and descendant generations were used in a field study of the inheritance of tolerance to Verticillium wilt, caused by Verticillium dahliae Kleb., in upland cotton (Gossypium hirsutum L.). The tolerant cultivar ‘Acala SJC-1’ was crossed to more susceptible parents, breeding line S5971 and cultivars ‘Acala 4-42’ and ‘Deltapine 70’. Seven generations were evaluated for each cross: the two parents (P1 and P2), F1; F2, F3, and reciprocal backcrosses (B1 and B2). The genetic control of tolerance in these crosses appears to involve more than one gene, based on an unsatisfactory fit to expected phenotypic distributions for the generations under a single-locus model. An analysis of generation means indicated that pooled additive and pooled dominance effects over loci were adequate to explain the variation among generations for crosses of SJC-1 X S5971 and SJC-1 X DPL70. Tolerance in these crosses appeared to be controlled by recessive factors. For the SJC-1 X 4-42 cross, an adequate fit to a digenic epistatic model was not possible, and none of the genetic parameters except the F2 mean were significant. Heritabilities for tolerance to Verticillium wilt, determined from regressions of F3 progeny on F2 parents for the crosses of SJC-1 X S5971 and SJC-1 X DPL70, ranged from 0.12 to 0.28. Therefore, individual plant selection for improved tolerance is expected to be inefficient.
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  • 3
    Electronic Resource
    Electronic Resource
    Springer
    Theoretical and applied genetics 84 (1992), S. 49-56 
    ISSN: 1432-2242
    Keywords: Linkage mapping ; Citrus ; Poncirus ; Molecular markers
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Isozymes and restriction fragment length polymorphisms were used as markers in the construction of a genetic map of the citrus nuclear genome. The map was based on the segregation of 8 isozyme, 1 protein, and 37 RFLP loci in 60 progeny of a cross of two intergeneric hybrids, ‘Sacaton’ citrumelo (Citrus paradisi Macf. x Poncirus trifoliata (L.) Raf.) and ‘Troyer’ citrange (C. sinensis (L.) Osbeck x P. trifoliata), often used as rootstocks. The map contains 38 of 46 studied loci distributed on ten linkage groups. A genome size of 1,700 cM was estimated from partial linkage data. Approximately 35% of the genome should be within 10 cM and 58% within 20 cM of the mapped markers. Eight loci in three linkage groups and 1 unlinked locus deviated significantly from Mendelian segregation.
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  • 4
    Electronic Resource
    Electronic Resource
    Springer
    Theoretical and applied genetics 94 (1997), S. 701-706 
    ISSN: 1432-2242
    Keywords: Key words Citrus ; Microsatellites ; STS markers ; SSLP ; SSR ; Null alleles
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract  We report the successful assignment of the first seven microsatellite markers to the Citrus RFLP and isozyme map. A total of 14 microsatellite primer pairs were developed and tested for amplification and product-length polymorphism within a population of plants previously used for linkage-map construction. In each case, the successfully assigned microsatellite mapped to the termini of a different linkage group indicating a widespread distribution throughout the genome. Analysis of allele segregation revealed that two of nine microsatellites displayed a significant deviation from expected ratios (P〉0.5). This was compared with other marker types within Citrus and a similar proportion of skewed loci was also found to be present. The analysis of two markers was complicated by the non-amplification of an inherited null allele within the mapping population. The successful integration of microsatellites into the genetic map of Citrus demonstrates the utility of this marker type for genetic analysis within wide intergeneric plant crosses.
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  • 5
    ISSN: 1432-2242
    Keywords: Key words Citrus ; RFLP ; RAPD ; Phylogeny ; Taxonomy
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract  Relationships among 88 accessions representing 45 Citrus species, three man-made hybrids, and six related genera were examined for restriction fragment length polymorphisms (RFLP). Thirty-two Citrus and three Microcitrus accessions were also examined by random amplified polymorphic DNA (RAPD) analysis. A measure of relative heterozygosity was estimated based on the mean of the number of fragments per individual per probe-enzyme combination (PEC) divided by total number of fragments per PEC for all non-hybrid Citrus individuals. The presence in a Citrus species of a rare band found also in a related genus was taken as an indication of possible introgression, while the presence of several fragments unique to 1 species was used to indicate non-involvement of that species in hybridization events. Most species that have been described in the literature as hybrids had high heterozygosity indices and no unique fragments. Distance matrices and dendrograms were generated using simple matching coefficient and neighbor-joining cluster analysis. RFLP and RAPD data gave approximately the same results. These data showed C. maxima was affiliated with the papedas C. hongheensis and C. latipes. C. medica clustered with C. indica when only non-hybrid taxa were examined, or among limes, lemons, and relatives when all species were considered. Mandarins did not show strongly supported groupings among themselves, nor with other species. These data showed that several accessions were probably assigned to the wrong species.
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  • 6
    ISSN: 1432-2242
    Keywords: Key words Poncirus trifoliata ; Trifoliate orange ; Isozymes ; RFLPs ; ISSR markers ; Fingerprinting
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract  Trifoliate orange [Poncirus trifoliata (L.) Raf.] is frequently used as a parent in citrus rootstock breeding, but the origin and amount of genetic diversity in germ plasm collections are poorly understood. Most accessions are self-compatible, but produce a mixture of sexual and apomictic seedlings. Variation among 48 vegetatively propagated trifoliate orange accessions was assessed at seven isozyme loci, together with the restriction fragment length polymorphisms (RFLPs) detected by 38 probe-enzyme combinations and the inter-simple sequence repeat (ISSR) markers generated by 11 primers. Isozymes and RFLPs detected few polymorphisms among accessions, although genetic analysis has shown that the common phenotype is heterozygous for four isozyme and at least four RFLP loci. ISSR amplification generated multiple banding profiles with an average of 58 fragments/primer/accession. These fragments were repeatable across DNA samples extracted from different trees of the same accession or extracted at different times, and across separate PCR runs. Seventeen unique marker phenotypes were identified. The 48 trifoliate orange accessions were classified into four major groups based on polymorphic ISSR markers. All large-flowered accessions are in group 4, while small-flowered accessions are in group 3. Many ISSR markers segregated in progeny derived by open-pollination (probably mostly selfing) of a common accession, indicating that these ISSR markers are also heterozygous. Accessions having identical genotypes for a large number of heterozygous markers are unlikely to have diverged by recombination. Thus the limited divergence we detected among most accessions most likely originated by mutation. ‘Monoembryonic’ and ‘Simmons’ differed from other accessions only in the loss of specific markers, indicating that they originated as zygotic seedlings of individuals similar to the common genotype. Three accessions recently introduced from China have relatively different fingerprints with 3–14 unique ISSR markers, and probably represent a much more divergent germ plasm that may be a valuable breeding resource.
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  • 7
    Electronic Resource
    Electronic Resource
    Springer
    Theoretical and applied genetics 95 (1997), S. 408-417 
    ISSN: 1432-2242
    Keywords: Key words Citrus ; Poncirus ; ISSR markers ; RFLPs ; Cultivar identification
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract  Inter-simple sequence repeat (ISSR) markers generated by 22 primers were tested for their ability to distinguish among samples from 94 trees of 68 citrus cultivars. Within each of the six cultivar groups studied, most of these cultivars are so closely related that they are difficult to distinguish by other molecular-marker techniques. ISSR markers involve PCR amplification of DNA using a single primer composed of a microsatellite sequence anchored at the 3′ or 5′ end by 2–4 arbitrary, often degenerate, nucleotides. The amplification products were separated on non-denaturing polyacrylamide gels and detected by silver staining. ISSR banding profiles were very repeatable on duplicate samples. Different citrus species had very different fingerprint patterns. Within Citrus sinensis (L.) Osbeck and C. paradisi Macf., in which all cultivars have originated by the selection of mutants, ISSR markers distinguished 14 of 33 sweet orange and 1 of 7 grapefruit cultivars. Five of six lemon cultivars were discriminated by ISSR markers. Many differences were found among mandarin cultivars; however, all five satsuma cultivars analyzed had identical ISSR fingerprints. Four of five citrange cultivars were distinguishable, but ‘Troyer’ and ‘Carrizo’ had identical ISSR fingerprints. ‘Kuharske Carrizo’ citrange, which has better citrus nematode resistance than other ‘Carrizo’ citrange accessions, had unique ISSR fingerprints. Three ISSR markers that differentiated certain sweet orange cultivars were hybridized to Southern blots of sweet orange DNA digested with different restriction endonucleases. The sweet orange cultivars tested could be distinguished by these ISSR-derived RFLP markers. Moreover, one ISSR marker unique to ‘Ruby’ blood orange was observed in its progeny trees.
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  • 8
    Electronic Resource
    Electronic Resource
    Springer
    Euphytica 92 (1996), S. 121-128 
    ISSN: 1573-5060
    Keywords: Lolium perenne ; perennial ryegrass ; co-adaptation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Summary Co-adaptation between neighbouring genotypes of Lolium perenne was examined in a pot experiment over an 18 month period. Competitive interactions in pure and mixed stands were examined using 9 genotype pairs (established as neighbours from electrophoresis data), 7 from a 10 year-old, and 2 from a 40+ year-old pasture. Combinations of each pair of neighbours grown together, and each grown with 5 randomly chosen alien (non-neighbour) genotypes sampled from the same pastures were also assessed. Seven harvests of green matter produced above 20 mm were made, and 50–135 kg N ha-1 was administered after each harvest. Data for dry weight at each harvest for pure and mixed stands, and Relative Yield Totals (RYT) were obtained for neighbours and aliens in each treatment. In 6 of the 9 pairs of neighbours grown as pure stands, the yields of neighbours did not differ significantly at the final harvest. In 8 of the 9 pairs grown as mixtures of pairs of neighbours, the yield of one of the neighbours was significantly reduced and one neighbour had been eliminated. Over the last 5 harvests, yields from (alien + neighbour) mixtures had the greatest dry matter yields. No suppression of one component by the other was recorded for this (alien + neighbour) combination. Based upon RYT estimates, neighbours did not yield more when grown together than when grown separately with aliens. There was no relationship between original genotype size in their fields of origin and dry matter yield in this experiment. This experiment provides no evidence for he evolution of co-adaptation between surviving neighbours in the two pastures from which these individuals were sampled. The genotypic architecture of pasture would seem to be the product of random and/or competitive exclusion of individual genotypes, rather than the evolution of co-adapted groups of individuals through time. The consequences of extreme depletion of genetic variation in the first two years after establishment, the very low rates of seedling establishment in established pastures, and the impact of the grazing animal would suggest that there is little likelihood of or potential for, co-adaptation evolving between neighbours.
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  • 9
    ISSN: 1573-4927
    Keywords: Tragopogon miscellus ; polyploidy ; alcohol dehydrogenase ; gene regulation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: Abstract This study demonstrates that homoeologous genes in two diploid plant species that specify different amounts of an enzyme maintain the same relative level of expression in an allotetraploid derivative. The three predominant alcohol dehydrogenase (ADH) isozymes (DD, DP, PP) in seeds of the recently evolved allotetraploid plant Tragopogon miscellus (Compositae) are dimers specified by Adh3-D and Adh3-P genes derived from its diploid progenitors T. dubius and T. pratensis. Seeds of T. pratensis contain twice as much ADH activity as those of T. dubius, while T. miscellus is intermediate. The three isozymes were similar in a number of catalytic properties; the densitometric ratio of the isozymes purified from T. miscellus was 1 DD:4DP:4PP for both ADH activity and protein; and dissociation-reassociation of the DP enzyme gave a 1:2:1 ratio of the three isozymes. Therefore, the enzymes were similar in specific activity, but twice as many P as D subunits were present in active enzymes in T. miscellus, precisely the difference in activity between the parents. In T. miscellus, the specific activity of ADH and its activity per mg tissue are intermediate to those of the diploids, because relative expression of the Adh gene in each genome is not influenced by the presence of the other genome.
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  • 10
    Electronic Resource
    Electronic Resource
    Springer
    Biochemical genetics 22 (1984), S. 631-643 
    ISSN: 1573-4927
    Keywords: Stephanomeria exigua ; alcohol dehydrogenase ; gene duplication ; isozymes
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: Abstract The alcohol dehydrogenase (ADH) isozymes induced in flooded roots of the diploid plant Stephanomeria exigua are specified by tightly linked genes comprising a complex locus, Adh1. Individuals homozygous for a complex with two active genes which specify electrophoretically different subunits have three ADH-I isozymes, two intragenic homodimers and an intergenic heterodimer. Individual isozymes were partially purified from plants homozygous for several different Adh1 complexes and apparent K m values for acetaldehyde, ethanol, NAD, and NADH and responses to temperature, pH, and two different alcohols were determined. The two homodimeric enzymes specified by a particular Adh1 complex generally differed in one or more of the properties studied, and in three of four cases, intergenic heterodimers differed significantly from intermediacy, often having lower K m values than either homodimer. None of the isozymes studied could be considered greatly divergent or defective. Constraints on evolution of duplicate genes which form intergenic heterodimers are considered.
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