Browsing by Author "W. Tushemereirwe"

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Developing a cell suspension system for Musa-AAA-EA cv. ‘Nakyetengu’: a critical step for genetic improvement of Matooke East African Highland bananas
(In Vitro Cellular & Developmental Biology - Plant, 2014-04-26) Priver Namanya; G. Mutumba; S. M. Magambo; W. Tushemereirwe
Embryogenic cell suspensions of triploid East African Highland bananas (Musa AAA-EA) were initiated and generated using cooking cultivar ‘Nakyetengu’ belonging to the Nakabululu clone set. Immature male flowers produced embryogenic calli consisting of embryos and friable tissue after 4 mo culture on a modified MA1 callus induction medium. Friable calli were initiated and maintained in liquid MA2 medium. A cell growth rate of 1.5–2.0 sedimented cell volume (SCV) per month was observed. Embryo development was observed at 2.18 × 103 embryos per mL SCV. Germination of these embryos was observed at 2.8% and 6.2% for two cell suspension lines. Plant regeneration efficiency was 60–100%, all producing normal plants with a shoot and roots at weaning. In the field, somatic cell-derived plants were all normal morphology and comparable to control plants during vegetative and reproductive stages. This study is a breakthrough for recalcitrant East African Highland banana and offers a system that can provide essential raw materials for associated germ- plasm improvement through genetic engineering approaches.
Developing A Cell Suspension System For Musa-Aaa-Ea Cv. ‘Nakyetengu’: A Critical Step For Genetic Improvement Of Matooke East African Highland Bananas
(Vitro Cellular & Developmental Biology - Plant, 2014-04-18) Priver Namanya; G. Mutumba; S. M. Magambo; W. Tushemereirwe
Embryogenic cell suspensions of triploid East African Highland bananas (Musa AAA-EA) were initiated and generated using cooking cultivar ‘Nakyetengu’ belonging to the Nakabululu clone set. Immature male flowers produced embryogenic calli consisting of embryos and friable tissue after 4 mo culture on a modified MA1 callus induction medium. Friable calli were initiated and maintained in liquid MA2 medium. A cell growth rate of 1.5–2.0 sedimented cell volume (SCV) per month was observed. Embryo development was observed at 2.18 × 103 embryos per mL SCV. Germination of these embryos was observed at 2.8% and 6.2% for two cell suspension lines. Plant regeneration efficiency was 60–100%, all producing normal plants with a shoot and roots at weaning. In the field, somatic cell-derived plants were all normal morphology and comparable to control plants during vegetative and reproductive stages. This study is a breakthrough for recalcitrant East African Highland banana and offers a system that can provide essential raw materials for associated germ- plasm improvement through genetic engineering approaches.
Nematode 18S rRNA gene is a reliable tool for environmental biosafety assessment of transgenic banana in confined field trials
(Transgenic Res, 2013-05-10) R. Nakacwa; A. Kiggundu; H. Talwana; J. Namaganda; C. Lilley; W. Tushemereirwe; H. Atkinson
Information on relatedness in nematodes is commonly obtained by DNA sequencing of the ribosomal internal transcribed spacer region. However, the level of diversity at this locus is often insufficient for reliable species differentiation. Recent findings suggest that the sequences of a fragment of the small subunit nuclear ribosomal DNA (18S rRNA or SSU), identify genera of soil nematodes and can also distinguish between species in some cases. A database of soil nematode genera in a Ugandan soil was developed using 18S rRNA sequences of individual nematodes from a GM banana confined field trial site at the National Agricultural Research Laboratories, Kawanda in Uganda. The trial was planted to evaluate transgenic bananas for resistance to black Sigatoka disease. Search for relatedness of the sequences gained with entries in a public genomic database identified a range of 20 different genera and sometimes distinguished species. Molecular markers were designed from the sequence information to underpin nematode faunal analysis. This approach provides bio-indicators for disturbance of the soil environment and the condition of the soil food web. It is being developed to support environmental biosafety analysis by detecting any perturbance by transgenic banana or other GM crops on the soil environment.