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  • 1
    ISSN: 1572-9788
    Keywords: Citrus ; seed ; seedless ; fruit
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract We have constructed chimaeric genes that are expressed in embryo and endosperm compartments of the seed, induce dominant seed lethality and have potential to reduce seed size in 75% of seeds within a fruit such as Citrus [7]. The genes are not entirely seed-specific as a proportion of primary test tobacco transformants containing their gene were fully male-sterile [7]. Here we investigated why a proportion of apparently male-fertile transgenic plants showed segregation distortion from the 75% seed lethality expected for a single dominant gene. Reciprocal crosses were conducted between pollen fertile, primary tobacco transformants containing various copies of the CG1-400-RNase gene [7] and wild-type tobacco plants to examine the transmission of the gene through maternal and paternal gametes and also the effects of gene dosage in embryo and endosperm compartments on seed viability and phenotype. Pollen viability, seed set and seed phenotype were examined over a 16 month period to assess stability of gene expression in primary transformants because woody, fruit crops containing these genes will be vegetatively propagated from primary transformants. In male-fertile transformants, the gene was observed to be expressed to varying degrees post-meiotically in pollen over the time period examined resulting in lethality of transgenic pollen and reduced paternal transmission. A variable, low-level maternal expression component was also detected that resulted in seed lethality and influenced morphological variation in the seed lethal phenotype. The maternal and paternal expression components caused seed lethality to range from 50 to 75%. This study indicates the need to select for transformants with stable pollen transmission and high seed expression and raises questions in relation to possible environmental and epistatic effects on gene expression in primary, hemizygous transformants over long growth periods.
    Type of Medium: Electronic Resource
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  • 2
    ISSN: 1572-9788
    Keywords: Citrus ; seed ; seedless ; fruit ; tobacco ; Arabidopsis
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract Seedlessness is a highly desirable characteristic in fresh fruit. Marketability of a fruit as seedless does not require complete absence of seeds as long as the seed structures are imperceptible during consumption. Chimaeric genes comprised of soybean β-conglycinin seed storage protein gene promoters linked to the bacterial RNase gene, Barnase, were tested for their efficacy to cause seed death and decrease seed size in tobacco and Arabidopsis. These species were used because they undergo two distinct seed developmental pathways and produce albuminous and exalbuminous seeds, respectively. In both species, the death of embryo and endosperm tissues occurred, resulting in a dominant seed lethal phenotype with segregation distortion. Reduction in seed size was only observed in Arabidopsis seeds and the phenotype resembled that of stenospermocarpic seeds in grape. Some transformants of both species were male-sterile and this correlated with the expression of the gene in anthers indicating that expression of the gene is not strictly seed-specific. The promoters also direct expression of a linked GUS gene to Citrus embryos of various developmental stages, and Citrus forms exalbuminous seeds, therefore, the Barnase constructions may be useful in eliciting a reduction in seed size of around 75% of the seeds found in the fruit. This may be sufficient to warrant marketing as ‘less seedy’ if trials in the cultivar of interest indicate that the smaller seeds are less detectable to the consumer. Abbreviations: GUS, β-glucuronidase; PCR, polymerase chain reaction; DMSO, dimethyl sulfoxide; DTA, diphtheria toxin-A chain; CFDA, 5(6)-carboxy-fluorescein di-acetate; CG, β-conglycinin; DAP, days after pollination; FAA, formaldehyde-acetic acid alcohol fixative.
    Type of Medium: Electronic Resource
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