48 research outputs found

    Some physiochemical properties of termite mound soil and its effect on yield and yield components of maize (Zea mays L.) Under Greenhouse condition at Nekemte, western Ethiopia

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    Termite mound soil amends soil fertility and utilized as an alternative to npk fertilizers by smallholder farmers in Africa. Experiment was conducted in western Ethiopia to compare selected physical and chemical properties of mound soil (ms), Adjacent soil (as) and non-mound soil (nms) and the effect of these soils on maize plant growth and yield in the Greenhouse. In Limu district, Fitbako kebele, cultivated land was purposively selected for sampling. In the cultivated land, a plot of 100m x100m was delineated and three mounds within the plot were also purposively selected to collect composite soil sample for soil analysis and Greenhouse pot experiment. From each mound, 10kg soil each from bottom, middle and top (total= 30 kg) were collected and mixed to obtain working sample of 10 kg. About 30kg of adjacent soil 5m away from mound soils and Non-mound soil at the distance of 20 m away from mound soil at the depth of 0-30cm were collected and thoroughly mixed to make working sample of 10kg from each soil type. Three kilograms of mound soil, adjacent soil and non-mound soil each was put in separate plastic bucket for greenhouse pot experiment. About 100 gm of each soil type was used for selected physical and chemical properties analyses. The result obtained demonstrated that termite mound soil was significantly (P<0.05) high in bulk density (bd), moisture contents (mc), porosity (P), Soil pH, percent organic carbon (% oc) and percent organic matter (% om). Total Nitrogen (tn), average (av.) P, av. K, Exch.  Ca and Exch. Mg were also significantly (P<0.05) higher in mound soil in comparison with adjacent soil and non-mound soil. Maize plant growth traits and yield were significantly (P<0.05) high in mound soil. From the current study, it can be concluded that the use of npk fertilizer on plots having termite mound is not recommended. However, further research is needed on how to use mound soil on large plot of lan

    Efficacy of botanical extracts against termites on maize (Zea mays (L.)) under field condition in western Ethiopia

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    Maize (Zea mays L.) is grown popularly in the world. It is severely attacked by termites. Insecticides are widely used to control, but could not minimize termites’ damage mainly because of resistance development and easily break down of the active ingredients. Hence, field experiment was conducted with the aim of evaluating different botanical crude extracts from leaves of Croton macrostachys (Hochst), Jatropha curcas L. and Phytolacca dodecandra L. for the management of termites.  The experiment was conducted at Wollega University (wu) under irrigation and rain fed conditions. Leaves of the botanicals were collected from experimental sites in western Ethiopia and dried under shade. The dried leaves were grounded to a fine powder using a small hand-operated manual grinder and the powder was further sliced with analytical mill and sieved through a 0.25 mm pore size mesh to make uniform fine dust particle. The treatments were C. macrostachys, P. dodecandra, J. curcas, C. macrostachys + P. dodecandra, C. macrostachys +J. curcas, P. dodecandra + J. curcas, C. macrostachys + P. dodecandra + J. curcas. Untreated check was used for comparison. For treatment preparation, 300 g. of each botanical leaf powder was soaked in 1000 ml distilled water for 72 hrs. The mixtures were filtered with clean cheese cloth and stored in a beaker of 250 ml capacity. The experiment was laid-out in a Randomized Complete Block Design (rcbd) in four replications. Shallow holes for maize seed planting were prepared well ahead of planting on the experimental plots. From the stock solution of each botanical, 20 ml was drenched to each planting hole 10 days before planting and continued at all maize growth stages. Treatments were applied using 30 ml capacity Syringe. Data on termite damage symptoms were collected two days before planting and two days after treatment application at every growth stage. The results revealed that mixed botanicals treatments were significantly (p<0.05) superior to non-mixed botanical treatments in the management of termites. The highest number of foraging termites, galleries and mounds were recorded in the untreated check plot, while the lowest was recorded in the mixed botanical treatments. Moreover, the highest number of maize stand count, maize cobs and maize yield were recorded from plots that received mixed botanicals and the lowest were recorded from the untreated plot. In conclusion, mixtures of C. macrostachys, J. curcas and P. dodecandra can be used as part of an integrated termites’ management

    Gene expression and plant performance in oryzacystatin-I expressing transformed tobacco (Nicotiana tabacum L. cv Samsun) plants under abiotic stress

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    Plant cysteine proteinase inhibitors or also called phytocystatins inhibit the action of cysteine proteinases in plants. These proteinases are involved in many developmental processes by degrading proteins. In this study possible effects of an exogenous oryzacystatin-I (OC-I) expressed in transformed tobacco has been investigated. By challenging OC-I expressing and non-expressing tobacco with drought and heat stress, OC-I transcription and translation were not affected in OC-I expressing plants and plant extracts from stressed plants containing the inhibitor inhibited papain activity in vitro. Further, plant growth and photosynthesis was not greatly different under the selected growth conditions in both plant types under stress and non-stress conditions. However, OC-I expressing plants showed slightly lower photosynthetic rate, were shorter and had a higher lower dry mass production under non-stress condition. By applying cDNA Representational Difference Analysis (cDNA-RDA) to detect differentially expressed genes in the two types of plants, a gene coding for the light harvesting chlorophyll a/b binding protein gene (lhcb1) of photosystem II (LHC II) was isolated from non-OCI expressing plants. Northern blot analysis showed lower transcript accumulation of the lhcb gene in OCI-expressing plants both under non-stress and stress conditions, which was accompanied by lower chlorophyll content in OC-I expressing plants. Furthermore, plants benefited from OC-I expression by protection of a variety of expressed proteins against degradation. Identification of possible target cysteine proteinases for OC-I in tobacco resulted in the isolation, cloning and characterization of two new papain-like cysteine proteinases from tobacco designated NtCP1 and NtCP2. NtCP1 was expressed only in senescent leaves and it was not induced in mature green leaves upon exposure to drought or heat stress. NtCP1 has therefore a possible potential as a developmental senescence marker in tobacco. In contrast, NtCP2, which was expressed in mature green leaves, has a high similarity to KDEL-tailed cysteine proteinases that are involved in programmed cell death. Both drought and heat decreased NtCP2 transcript abundance in mature green leaves. Overall, this study has provided evidence that expression of exogenous OC-I does not significantly improve plant performance in tobacco in terms of physiological traits under drought and heat stress but provides protection in terms of stability of protein expression by possibly interacting with endogenous tobacco cysteine proteinases. Further detailed studies are suggested on the interaction of endogenous cysteine proteinases and exogenous phytocystatins to elucidate in more detail the type of interaction. Copyright 2006, University of Pretoria. All rights reserved. The copyright in this work vests in the University of Pretoria. No part of this work may be reproduced or transmitted in any form or by any means, without the prior written permission of the University of Pretoria. Please cite as follows: Beyene, G 2006, Gene expression and plant performance in oryzacystatin-I expressing transformed tobacco (Nicotiana tabacum L. cv Samsun) plants under abiotic stress, PhD thesis, University of Pretoria, Pretoria, viewed yymmdd Thesis (PhD (Botany))--University of Pretoria, 2006.Plant Scienceunrestricte

    From Traditional Breeding to Genome Editing for Boosting Productivity of the Ancient Grain Tef Eragrostis tef (Zucc.) Trotter

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    Tef (Eragrostis tef (Zucc.) Trotter) is a staple food crop for 70 of the Ethiopian population and is currently cultivated in several countries for grain and forage production. It is one of the most nutritious grains, and is also more resilient to marginal soil and climate conditions than major cereals such as maize, wheat and rice. However, tef is an extremely low-yielding crop, mainly due to lodging, which is when stalks fall on the ground irreversibly, and prolonged drought during the growing season. Climate change is triggering several biotic and abiotic stresses which are expected to cause severe food shortages in the foreseeable future. This has necessitated an alternative and robust approach in order to improve resilience to diverse types of stresses and increase crop yields. Traditional breeding has been extensively implemented to develop crop varieties with traits of interest, although the technique has several limitations. Currently, genome editing technologies are receiving increased interest among plant biologists as a means of improving key agronomic traits. In this review, the potential application of clustered regularly interspaced short palindromic repeats (CRISPR) and CRISPR-associated proteins (CRISPR-Cas) technology in improving stress resilience in tef is discussed. Several putative abiotic stress-resilient genes of the related monocot plant species have been discussed and proposed as target genes for editing in tef through the CRISPR-Cas system. This is expected to improve stress resilience and boost productivity, thereby ensuring food and nutrition security in the region where it is needed the most

    Use of transgenic Oryzacystatin-I expressing plants enhances recombinant protein production

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    Plants are an effective and inexpensive host for the production of commercially interesting heterologous recombinant proteins. The Escherichia coli-derived glutathione reductase was transiently expressed as a recombinant model protein in the cytosol of tobacco plants using the technique of leaf agro-infiltration. Proteolytic cysteine protease activity progressively increased over time when glutathione reductase accumulated in leaves. Application of cysteine protease promoter–GUS fusions in transgenic tobacco identified a cysteine protease NtCP2 expressed in mature leaves and being stress responsive to be expressed as a consequence of agro-infiltration. Transgenic tobacco plants constitutively expressing the rice cysteine protease inhibitor oryzacystatin-I had significantly lower cysteine protease activity when compared to non-transgenic tobacco plants. Lower cysteine protease activity in transgenic plants was directly related to higher glutathione reductase activity and also higher glutathione reductase amounts in transgenic plants. Overall, our work has demonstrated as a novel aspect that transgenic tobacco plants constitutively expressing an exogenous cysteine protease inhibitor have the potential for producing more recombinant protein which is very likely due to the reduced activity of endogenous cysteine protease.This work was supported by a grant from the National Research Foundation in South Africa.http://link.springer.com/journal/12010hb201

    Provitamin A biofortification of cassava enhances shelf life but reduces dry matter content of storage roots due to altered carbon partitioning into starch

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    Storage roots of cassava (Manihot esculenta Crantz), a major subsistence crop of sub-Saharan Africa, are calorie rich but deficient in essential micronutrients, including provitamin A β-carotene. In this study, β-carotene concentrations in cassava storage roots were enhanced by coexpression of transgenes for deoxy-d-xylulose-5-phosphate synthase (DXS) and bacterial phytoene synthase (crtB), mediated by the patatin-type 1 promoter. Storage roots harvested from field-grown plants accumulated carotenoids to ≤50 lg/g DW, 15- to 20-fold increases relative to roots from nontransgenic plants. Approximately 85%–90% of these carotenoids accumulated as all-trans-β-carotene, the most nutritionally efficacious carotenoid. β-Carotene-accumulating storage roots displayed delayed onset of postharvest physiological deterioration, a major constraint limiting utilization of cassava products. Large metabolite changes were detected in β-carotene-enhanced storage roots. Most significantly, an inverse correlation was observed between β-carotene and dry matter content, with reductions of 50%–60% of dry matter content in the highest carotenoid-accumulating storage roots of different cultivars. Further analysis confirmed a concomitant reduction in starch content and increased levels of total fatty acids, triacylglycerols, soluble sugars and abscisic acid. Potato engineered to co-express DXS and crtB displayed a similar correlation between β-carotene accumulation, reduced dry matter and starch content and elevated oil and soluble sugars in tubers. Transcriptome analyses revealed a reduced expression of genes involved in starch biosynthesis including ADP-glucose pyrophosphorylase genes in transgenic, carotene-accumulating cassava roots relative to nontransgenic roots. These findings highlight unintended metabolic consequences of provitamin A biofortification of starch-rich organs and point to strategies for redirecting metabolic flux to restore starch production

    Mutations in DNA polymerase δ subunit 1 co-segregate with CMD2-type resistance to Cassava Mosaic Geminiviruses

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    Cassava mosaic disease (CMD) suppresses cassava yields across the tropics. The dominant CMD2 locus confers resistance to cassava mosaic geminiviruses. It has been reported that CMD2-type landraces lose resistance after regeneration through de novo morphogenesis. As full genome bisulfite sequencing failed to uncover an epigenetic mechanism for this loss of resistance, whole genome sequencing and genetic variant analysis was performed and the CMD2 locus was fine-mapped to a 190 kilobase interval. Collectively, these data indicate that CMD2-type resistance is caused by a nonsynonymous, single nucleotide polymorphism in DNA polymerase δ subunit 1 (MePOLD1) located within this region. Virus-induced gene silencing of MePOLD1 in a CMD-susceptible cassava variety produced a recovery phenotype typical of CMD2-type resistance. Analysis of other CMD2-type cassava varieties identified additional candidate resistance alleles within MePOLD1. Genetic variation of MePOLD1, therefore, could represent an important genetic resource for resistance breeding and/or genome editing, and elucidating mechanisms of resistance to geminiviruses

    Enhanced Transgene Expression in Sugarcane by Co-Expression of Virus-Encoded RNA Silencing Suppressors

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    Post-transcriptional gene silencing is commonly observed in polyploid species and often poses a major limitation to plant improvement via biotechnology. Five plant viral suppressors of RNA silencing were evaluated for their ability to counteract gene silencing and enhance the expression of the Enhanced Yellow Fluorescent Protein (EYFP) or the β-glucuronidase (GUS) reporter gene in sugarcane, a major sugar and biomass producing polyploid. Functionality of these suppressors was first verified in Nicotiana benthamiana and onion epidermal cells, and later tested by transient expression in sugarcane young leaf segments and protoplasts. In young leaf segments co-expressing a suppressor, EYFP reached its maximum expression at 48-96 h post-DNA introduction and maintained its peak expression for a longer time compared with that in the absence of a suppressor. Among the five suppressors, Tomato bushy stunt virus-encoded P19 and Barley stripe mosaic virus-encoded γb were the most efficient. Co-expression with P19 and γb enhanced EYFP expression 4.6-fold and 3.6-fold in young leaf segments, and GUS activity 2.3-fold and 2.4-fold in protoplasts compared with those in the absence of a suppressor, respectively. In transgenic sugarcane, co-expression of GUS and P19 suppressor showed the highest accumulation of GUS levels with an average of 2.7-fold more than when GUS was expressed alone, with no detrimental phenotypic effects. The two established transient expression assays, based on young leaf segments and protoplasts, and confirmed by stable transgene expression, offer a rapid versatile system to verify the efficiency of RNA silencing suppressors that proved to be valuable in enhancing and stabilizing transgene expression in sugarcane

    Multiple morphogenic culture systems cause loss of resistance to cassava mosaic disease

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    Abstract Background Morphogenic culture systems are central to crop improvement programs that utilize transgenic and genome editing technologies. We previously reported that CMD2-type cassava (Manihot esculenta) cultivars lose resistance to cassava mosaic disease (CMD) when passed through somatic embryogenesis. As a result, these plants cannot be developed as products for deployment where CMD is endemic such as sub-Saharan Africa or the Indian sub-continent. Result In order to increase understanding of this phenomenon, 21 African cassava cultivars were screened for resistance to CMD after regeneration through somatic embryogenesis. Fifteen cultivars were shown to retain resistance to CMD through somatic embryogenesis, confirming that the existing transformation and gene editing systems can be employed in these genetic backgrounds without compromising resistance to geminivirus infection. CMD2-type cultivars were also subjected to plant regeneration via caulogenesis and meristem tip culture, resulting in 25–36% and 5–10% of regenerated plant lines losing resistance to CMD respectively. Conclusions This study provides clear evidence that multiple morphogenic systems can result in loss of resistance to CMD, and that somatic embryogenesis per se is not the underlying cause of this phenomenon. The information described here is critical for interpreting genomic, transcriptomic and epigenomic datasets aimed at understanding CMD resistance mechanisms in cassava
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