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    Transformation and Regeneration Protocol for Two Farmer Preferred Open Pollinated Tropical Maize (Zea Mays) Varieties

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    U16ArtNeondoTransformationNothomDev.pdf (3.373Mb)
    Date
    2016-08-03
    Author
    Neondo, J.O.
    Alakonya, A.E.
    Matheka, J.
    Masanga, J.O.
    Kasili, R.W.
    Type
    Journal Article
    Target Audience
    Scientists
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    Abstract/Description
    Abstract: In vitro regeneration of open pollinated varieties (OPVs) Kakamega Striga Tolerant Population 94 (KSTP’94) and ‘Namba Nane’ alongside a tropical inbred line (CML144) was evaluated using immature zygotic embryos as explants. Four callus induction media (CIM) regimes; Murashige and Skoog (MS), Linsmaier and Skoog (LS), Chu (N6) and N6*(N6 medium fortified with 0.35 gL-1 L-proline and 0.8 mgL-1 AgNO3) were evaluated for their potential to induce callus in the three genotypes. All the media were supplemented with sucrose and five levels of 2, 4-Dichlorophenoxyacetic acid (2, 4-D) (0.5, 1.0, 1.5, 2.0 and 2.5 mgL-1). Resulting calli were matured on MS and N6 basal media supplemented with 60 g/L sucrose and similar concentration levels (0.5, 1.0, 1.5, 2.0 and 2.5 mgL-1) of 2, 4-D while the subsequent embryogenic calli were regenerated on hormone-free media. Transformability of these varieties was assessed via histochemical analysis of β-glucuronidase (GUS) reporter gene following Agrobacterium-mediated transformation. Statistical analyses were done using Statistical Analysis Software (SAS) and Graphpad Prism softwares with mean separations achieved at 95% confidence intervals. Of the 2 OPVs, KSTP’94 recorded the highest callus induction frequency (84.4%) while Namba Nane (45.6%) had the lowest. Similarly, KSTP, 94 had the highest mean of mature somatic embryos (59.7%) while Namba Nane recorded the lowest (16.4%). Assessment of regeneration frequencies from embryogenic calli revealed no significant differences among the 3 lines although CML 144 had the highest mean number of juvenile plantlets (36.7%). Analysis of transformation frequency (upon selection of calli on media with basta) showed that Namba Nane recorded the lowest transformation frequency (average 13.5%) some words missing. Transformation frequency (based on GUS positive calli) of these varieties ranged from 0.8 to 2.1%. This work therefore provides an empirical platform for potential introduction of useful genes into these varieties.
    https://dx.doi.org/10.11648/j.jps.20160404.14
    Multi standard citation
    Permanent link to this item
    https://hdl.handle.net/20.500.12478/1248
    Digital Object Identifier (DOI)
    https://dx.doi.org/10.11648/j.jps.20160404.14
    IITA Subjects
    Maize
    Agrovoc Terms
    Embryogenesis; Maize; Regeneration; Open Pollination; Callus
    Regions
    Africa; East Africa
    Countries
    Kenya
    Journals
    Journal of Plant Sciences
    Collections
    • Journal and Journal Articles4835
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