Regeneration of Sudanese maize inbred lines and open pollinated varieties

Omer, Rasha Adam; Ali, Asif; Matheka, Jonathan; Machuka, Jesse

doi:10.5897/ajb08.856

Cited by 11 publications

(3 citation statements)

References 43 publications

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“…The latter PGR caused precocious germination and produced nonembryogenic calli. Previous studies have also implicated 2,4-D in negative somaclonal variations in regenerants (Akoyi et al, 2013;Omer et al, 2008).…”

Section: Discussionmentioning

confidence: 87%

“…Efficient use of genetic engineering techniques in the improvement of maize is hampered by the in vitro recalcitrance frequently seen in most cereals making it difficult to induce embryogenic callus and eventual regeneration of maize especially in elite varieties (Ge et al, 2016). Despite this challenge, transformation and regeneration of tropical maize lines and other genotypes including open pollinated varieties and hybrids (Ombori et al, 2008(Ombori et al, , 2013 has been achieved, albeit with some difficulty (Muoma et al, 2008;Oduor et al, 2006;Ombori et al, 2008;Omer et al, 2008). Successful transformation utilizes immature zygotic embryos as explants for indirect transgene delivery often mediated by Agrobacterium tumefaciens (Akoyi et al, 2013;Anami et al, 2010;Mgutu et al, 2011;Ombori et al, 2008).…”

Section: Introductionmentioning

confidence: 99%

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An optimized protocol for in vitro regeneration of tropical maize inbred lines through cell suspension and semi-protoplast cultures

Billiah,

Joel,

Steven

2023

Afr. J. Biotechnol.

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Maize being an important staple food crop is widely consumed in Kenya but its production remains low due to biotic and abiotic challenges that have not been addressed sufficiently through conventional breeding. This study sought to optimize the establishment of cell suspension and protoplast cultures for 10 tropical inbred maize genotypes in order to identify genotypes that can readily establish in liquid cultures and those whose cell suspensions and protoplasts are capable of in vitro regeneration. Callus were induced using immature zygotic embryos on Murashige and Skoog (MS) medium supplemented with 3 mg/L 2, 4-D, 5 mg/L dicamba or 5 mg/L picloram. Dicamba and picloram induced friable calli that readily dispersed into cell suspensions in liquid MS medium supplemented with 0.1 g/L asparagine and either 0.4 or 0.8 g/L proline. Cell growth was determined by packed cell volume (PCV) every seven days. The highest PCV (240 µl/ml) was recorded in genotype EO4 followed by CML 216 (188 µ/ml). Optimal growth was observed in cells maintained in MS Amended with 0.4 or 0.8 g/L of proline in combination with 0.1 g/L asparagine compared to medium without proline. It was also observed that cells cultured in media with reduced Ammonium Nitrate (12 fold reduction) recorded higher PCV values than controls. Protoplasts were generated from the resulting cells using 2% cellulase and 0.5% pectolyase in an enzyme digestion cocktail containing Mannitol and Calcium Chloride (MaCa) and washed in MS with vitamins containing Mannitol (MSMa). Only cell clusters of genotype EO4 gave rise to plants with a regeneration frequency of 42.51%. In conclusion, success was achieved in callus initiation, formation of cell suspension cultures and their eventual regeneration into whole plants for selected tropical maize genotypes.

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Section: Discussionmentioning

confidence: 87%

Section: Introductionmentioning

confidence: 99%

An optimized protocol for in vitro regeneration of tropical maize inbred lines through cell suspension and semi-protoplast cultures

Billiah,

Joel,

Steven

2023

Afr. J. Biotechnol.

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“…DNA isolated by the Dolye method (Ba et al, 2004;Tosti et al, 2005;Sarutayophat et al, 2007), the Dellaporta method (Fatokun et al, 1992;Myers et al, 1996), the Rogers method (Zannou et al, 2008;Malviya et al, 2012), the Saghai-Maroof method (Tantssawat et al, 2010) and the commercial kit (Ghalmi et al, 2010;Fang et al, 2007) have been already used in the genetic analysis of cowpea. However, there is a lack of comparative research of these DNA extraction methods.…”

Section: Introductionmentioning

confidence: 99%

Comparative Analysis of Six DNA Extraction Methods in Cowpea (Vigna unguiculata L.Walp)

Tan¹,

Huang²,

Tie³

et al. 2013

JAS

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High quality DNA extractions are a prerequisite for genetic studies of a variety of plants including cowpea (Vigna unguiculata). Nowadays, there are a great number of plant DNA extraction methods, and commercially available extraction kits are also becoming more and more popular. It appears that different procedures work best for different plant groups. Thus in the genetic studies of cowpea, which DNA extraction method to choose becomes a concern. To solve this problem, five classic plant DNA isolation methods, including three CTAB methods and two SDS methods, were compared and evaluated while isolation using a commercial kit was also undertaken. The DNA extracted by these six methods from two-week-old cowpea seedlings were analyzed according to their cost and time, yield, purity, integrity, and functionality in restriction endonuclease digestion and PCR (polymerase chain reaction) based downstream analysis. After the evaluation, one most suitable method, described by Dellaporta et al. (1983) was selected and chosen for isolating DNA from young leaves of cowpea seedlings. The cost and time required in this method was relatively low. In addition, the quantity and the quality of the DNA extracted by this method were high enough to perform hundreds of PCR-based reactions.

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