Silas D. Obukosia

Elevated vitamin E content improves all-trans β-carotene accumulation and stability in biofortified sorghum

Significance Studies on the importance of vitamin A for human health continue to draw significant worldwide attention. However, the instability of provitamin A in crops resulted in a significant reduction of the potential nutrition values of these food crops. Our work demonstrates that provitamin A can be stabilized in sorghum by the coexpression of vitamin E through ectopic expression of homogentisate geranylgeranyltransferase (HGGT) and that vitamin E can enhance the stability of provitamin A in planta. This research has the potential to impact directly the lives of the millions of people who suffer from vitamin A deficiency, and we believe that these results will be applicable to enhancing provitamin A stability in many food crops. Micronutrient deficiencies are common in locales where people must rely upon sorghum as their staple diet. Sorghum grain is seriously deficient in provitamin A (β-carotene) and in the bioavailability of iron and zinc. Biofortification is a process to improve crops for one or more micronutrient deficiencies. We have developed sorghum with increased β-carotene accumulation that will alleviate vitamin A deficiency among people who rely on sorghum as their dietary staple. However, subsequent β-carotene instability during storage negatively affects the full utilization of this essential micronutrient. We determined that oxidation is the main factor causing β-carotene degradation under ambient conditions. We further demonstrated that coexpression of homogentisate geranylgeranyl transferase (HGGT), stacked with carotenoid biosynthesis genes, can mitigate β-carotene oxidative degradation, resulting in increased β-carotene accumulation and stability. A kinetic study of β-carotene degradation showed that the half-life of β-carotene is extended from less than 4 wk to 10 wk on average with HGGT coexpression.

EFFECTS OF GROWTH REGULATORS AND GENOTYPES ON PYRETHRUM IN VITRO

SummaryThe effects of growth regulators on culture response of different pyrethrum (Chrysanthemum cinerariaefolium Vis.) genotypes were investigated. In the genotype Sb/66/107, the presence of 2,4-dichorophenoxyacetic acid (2,4-D) at 2 mgl−1 promoted growth of callus, whereas benzyladenine and 1-naphthaleneacetic acid had no effect. Callus growth was also affected by the 2,4-D, ranging from 0.5 mgl−1 for genotype Marwanga to 3.0 mgl−1 for Ks/75/336. Among the genotypes, shoots were regenerated from calluses of Sb/66/107, 4331, Marwanga, and MA/70/1013.

Expression of plastid-encoded photosynthetic genes during chloroplast or chromoplast differentiation in Cucurbitae pepo L. fruits.

The objective of the study was to determine the patterns of expression of two photosynthetic genes rbcL and psbA, during chloroplast and chromoplast differentiation in fruit tissues of three Cucurbitae pepo L. cultivars: Early Prolific, Foodhook Zucchini and Bicolor Gourds. In two Early Prolific isogenic lines, YYBB and YYB+B+, the steady-state amounts of rbcL and psbA transcripts increased with fruit development upto 14 days post-pollination. The YYB+B+ line in which chloroplast differentiates into chromoplast at about pollination, did not show significantly higher amounts of both transcripts compared to YYBB, in which chromoplast develops early prior to pollination. In the Bicolor Gourds, in which the chromoplast and chloroplast containing tissues lie in juxtaposition on the same fruit, showed little differences in rbcL and psbA transcripts between the two tissues, if any the chromoplast containing tissue contained more of both transcripts than the chloroplast containing tissue. In Fordhook Zucchini fruits, where the chloroplast containing tissue developed early prior to pollination and was maintained, the steady-state amounts of rbcL transcripts increased to a maximum at 3 days post-pollination and levelled at 14 and 21 days post-pollination. In contrast, in Fordhook Zucchini fruits, the psbA transcript increased gradually up to 21 days post-pollination. In Fordhook Zucchini, the apparent ratios of psbA transcripts versus rbcL transcripts ranged from 2.5 to 3.9, at day 3 to 21 post-pollination, while in Bicolor Gourds were 2.9 and 4.5 at days 14 and 21 post-pollination. The two photosynthetic genes, psbA and rbcL were developmentally regulated and differentially expressed. However, their expression in chloroplast containing fruit tissues was not higher than in the chromoplast containing fruit tissues.

Is there a place for nutrition-sensitive agriculture?

The focus of the review paper is to discuss how biotechnological innovations are opening new frontiers to mitigate nutrition in key agricultural crops with potential for large-scale health impact to people in Africa. The general objective of the Africa Biofortified Sorghum (ABS) project is to develop and deploy sorghum with enhanced pro-vitamin A to farmers and end-users in Africa to alleviate vitamin A-related micronutrient deficiency diseases. To achieve this objective the project technology development team has developed several promising high pro-vitamin A sorghum events. ABS 203 events are so far the most advanced and well-characterised lead events with about 12 μg β-carotene/g tissue which would supply about 40-50 % of the daily recommended vitamin A at harvest. Through gene expression optimisation other events with higher amounts of pro-vitamin A, including ABS 214, ABS 235, ABS 239 with 25, 30-40, 40-50 μg β-carotene/g tissue, respectively, have been developed. ABS 239 would provide twice recommended pro-vitamin A at harvest, 50-90 % after 3 months storage and 13-45 % after 6 months storage for children. Preliminary results of introgression of ABS pro-vitamin A traits into local sorghum varieties in target countries Nigeria and Kenya show stable introgression of ABS vitamin A into local farmer-preferred sorghums varieties. ABS gene Intellectual Property Rights and Freedom to Operate have been donated for use royalty free for Africa. Prior to the focus on the current target countries, the project was implemented by fourteen institutions in Africa and the USA. For the next 5 years, the project will complete ABS product development, complete regulatory science data package and apply for product deregulation in target African countries.

The population structure of wild sorghum species in agro-ecological zones of Western Kenya

There is need to understand the genetic structure of wild sorghums that grow alongside cultivated traditional sorghum varieties in order to assess the potential effect of crop genes in wild populations. In this study, 175 wild sorghum samples were collected from 13 agroecological zones (AEZs) from three counties in Western Kenya and genotyped using microsattelite markers. Crop alleles were observed in wild sorghum populations. The range of allelic frequencies varied from low (˂0.4), to moderate (0.4-0.7) and to high (0.7) in the AEZs. Wild sorghum populations had moderate to high expected heterozygosity (HE) values of between 0.453 in LM1 to 0.715 in LM2. Differences in the magnitude of diversity was significant in the counties (Busia HE = 0.59 – 0.71; Homabay HE = 0.58-0.68 and Siaya HE = 0.45-0.59) but not distinct among the AEZs. Whole population FIS, FST and FIT values were low at 0.15, 0.16 and 0.29, respectively indicating low level of inbreeding, low genetic differentiation of the population and low to moderate deviation from Hardy–Weinberg (HW) equilibrium respectively. The deviation from HW equilibrium was significant in some wild populations from Siaya and Busia. Intra-population diversity (HS) was larger than inter-population diversity (DST) in 13 populations from the sampled AEZs, indicating the importance of gene flow between populations of wild sorghums. Heterozygosity values under mutation drift equilibrium (HEQ) varied under infinite allele model (IAM), two–phase model (TPM) and the step wise mutation model (SMM). However, significant population bottlenecks were absent in the wild sorghums. Presence of significant geographic county clusters and lack of significance on AEZ clusters indicate that human activities have had more influence on the distribution and diversity of wild sorghums than the prevailing climatic conditions. Efforts towards physical and genetic containment of crops genes need to be enhanced for successful ecologically sensitive confined field trials and future adoption of transgenics in cropping systems. Keywords: Diversity, Sorghum bicolor , Sorghum halepense , Sorghum sudanense , microsatellite loci. Abbreviation: AEZ, Agro-ecological zone; DST, inter-population gene diversity; FIS, fixation index; FIT, index of deviation from HW equilibrium; FST, degree of population differentiation; GST, proportion of inter-population gene diversity; HE, expected heterozygosity; HEQ, heterozygosity values under mutation drift equilibrium; HO, observed heterozygosity; HT, total gene diversity; IAM, infinite allele model; SMM, stepwise mutation model; TPM, two-phase model; LM, lower Midlands; UM, upper midlands; HB, Homabay; SY, Siaya; BU, Busia counties; SSR, simple sequence repeats.

Effects of Genotype and in Vitro Media Composition on Micropropagation of Sugar Cane

The use of plant tissue culture for plant regeneration and propagation on large-scale requires a quantitative approach to optimised conditions associated with vegetative plant regeneration (Lal and Lal, 1990). Numerous factors affect morphogenesis and proliferation rates in micropropagation systems. These include the media composition (Nitsch, 1969; Huang and Murashige, 1977); the nature of donor and explant (Gukasyan et al., 1977; Wang and Ma, 1978); light (Hasegawa et al., 1973); temperature (Carew and Staba, 1965; Seabrook and Cumming, 1978); polarity (Blotch, 1943, Ziv et al., 1970); gas phase (Beasely and Eaks, 1979; Street, 1979), subculture (Hill, 1967; Chen and Galstron, 1967); genotype (Pierik et al., 1974; Pierik and Steegmans, 1976; Seabrook and Cumming, 1978) and season (Robb, 1957; Nitsch and Hughes, 1978). In order to optimise plant regeneration and propagation, a wide range of factors should be investigated.