scholarly journals Efficient Genetic Transformation and Regeneration of a Farmer-Preferred Cassava Cultivar From Ghana

2021 ◽  
Vol 12 ◽  
Author(s):  
Wilfred Elegba ◽  
Emily McCallum ◽  
Wilhelm Gruissem ◽  
Hervé Vanderschuren

Cassava is an important staple crop that provides food and income for about 700 million Africans. Cassava productivity in Africa is limited by viral diseases, mainly cassava mosaic disease (CMD) and cassava brown streak disease (CBSD). Genetic barriers such as high heterozygosity, allopolyploidy, poor seed set, and irregular flowering constrain the development of virus-resistant cassava varieties via conventional breeding. Genetic transformation represents a valuable tool to circumvent several challenges associated with the development of virus resistance and other valuable agronomic traits in cassava. The implementation of genetic transformation in many local African cultivars is limited either by the difficulty to produce friable embryogenic callus (FEC), low transformation, and/or regeneration efficiencies. Here, we report the successful induction of organized embryogenic structures (OES) in 11 farmer-preferred cultivars locally grown in Ghana. The production of high quality FEC from one local cultivar, ADI 001, facilitated its genetic transformation with high shoot regeneration and selection efficiency, comparable to the model cassava cultivar 60444. We show that using flow cytometry for analysis of nuclear ploidy in FEC tissues prior to genetic transformation ensures the selection of genetically uniform FEC tissue for transformation. The high percentage of single insertion events in transgenic lines indicates the suitability of the ADI 001 cultivar for the introduction of virus resistance and other useful agronomic traits into the farmer-preferred cassava germplasm in Ghana and Africa.

2018 ◽  
Vol 10 (12) ◽  
pp. 78
Author(s):  
Eric Mpongolo Musungayi ◽  
Kahiu Ngugi ◽  
James Wanjohi Muthomi ◽  
Vincent Woyengo Were ◽  
Florence Mmogi Olubayo ◽  
...  

Cassava mosaic disease (CMD) caused by Bemisia tabaci is among the major contributors to low cassava yield in Africa and therefore requires instituting control measures. Due to genetic diversity in cassava, only clones with superior agronomic traits, disease resistance and high yield are selected and released to farmers or deployed in breeding program. This study was conducted to evaluate the resistance of cassava half-sib families to CMD. Field trials were conducted at Kenya Agricultural and Livestock Research Organization (KALRO), at Kakamega and Alupe research stations in western Kenya from June 2016 to June 2017. Sixty progenies were compared to that of their five parents by planting cuttings in 4 × 2 meters plots. Data were collected on plant height, number of roots per plant, harvest index, root yield, dry matter content, cassava mosaic disease and whiteflies infestation. Twenty three genotypes had a mean score of 1.0 to CMD, implying that they are resistant. Cassava grown at Alupe was observed to have high number of susceptible genotypes compared to cassava grown at Kakamega, indicating the effect of the environment on the genotypes. Parental genotypes, Kaleso and MM96/4271 presented high number of progenies showing CMD resistance. Genotypes, P4G1 and P2G3 with mean root yield of 31.6 t ha-1 and 30.0 t ha-1 were the highest yielding in term of root yield. A number of half-sib families generated from MM96/4271, Kaleso and MM96/0686 performed well with respect to yield recorded on their respective parents. Evaluation of new cassava varieties under local disease conditions would most likely improve the productivity of cassava through selection of resistant clones.


2016 ◽  
Vol 07 (07) ◽  
pp. 1122-1128 ◽  
Author(s):  
Patrick Chiza Chikoti ◽  
Paul Shanahan ◽  
Rob Melis

2021 ◽  
Vol 12 ◽  
Author(s):  
Chukwuka Ugochukwu Ano ◽  
Mildred Ochwo-Ssemakula ◽  
Angele Ibanda ◽  
Alfred Ozimati ◽  
Paul Gibson ◽  
...  

Cassava mosaic geminiviruses (CMGs) and cassava brown streak viruses (CBSVs) cause the highest yield losses in cassava production in Africa. In particular, cassava brown streak disease (CBSD) is and continues to be a significant constraint to optimal cassava production in Eastern and Southern Africa. While CBSD has not been reported in West Africa, its recent rapid spread and damage to cassava productivity in Eastern, and Southern Africa is alarming. The aim of this study was to evaluate Nigerian cassava genotypes in order to determine their responses to CBSD, in the event that it invades Nigeria, the world’s largest cassava producer. The study gathered information on whether useful CBSD resistance alleles are present in the elite Nigerian cassava accessions. A total of 1,980 full-sib cassava seedlings from 106 families were assessed in the field at the seedling stage for a year. A subset of 569 clones were selected and assessed for another year at the clonal stage in Namulonge, central Uganda, a known hotspot for CBSD screening. Results indicated that foliar and root incidences and severities varied significantly (p ≤ 0.01, p ≤ 0.001) except for CBSD foliar incidence at 6 months (CBSD6i). Highest and lowest plot-based heritability estimates for CBSD were registered for CBSD root severity (CBSDrs) (0.71) and CBSD6i (0.5). Positive and highly significant correlations were noted between CBSD root incidence (CBSDri) and CBSDrs (r = 0.90***). Significant positive correlations were also noted between CBSD foliar severity at 3 months (CBSD3s) and CBSD foliar incidence at 6 months (CBSD6i) (r = 0.77***), CBSD3s and CBSDrs (r = 0.35***). Fresh root weight (FreshRW) negatively correlated with CBSDri and CBSDrs, respectively (r = −0.21*** and r = −0.22***). Similarly, CBSD3s correlated negatively with cassava mosaic disease severity at 3 (CMD3s) and 6 months (CMD6s), respectively (r = −0.25*** and r = −0.21***). Fifteen clones were selected using a non-weighted summation selection index for further screening. In conclusion, results revealed that the elite Nigerian accessions exhibited significant susceptibility to CBSD within 2 years of evaluation period. It is expected that this information will aid future breeding decisions for the improvement of CBSD resistance among the Nigerian cassava varieties.


2021 ◽  
Vol 83 (8) ◽  
Author(s):  
F. Al Basir ◽  
Y. N. Kyrychko ◽  
K. B. Blyuss ◽  
S. Ray

AbstractMany plant diseases are caused by plant viruses that are often transmitted to plants by vectors. For instance, the cassava mosaic disease, which is spread by whiteflies, has a significant negative effect on plant growth and development. Since only mature whiteflies can contribute to the spread of the cassava mosaic virus, and the maturation time is non-negligible compared to whitefly lifetime, it is important to consider the effects this maturation time can have on the dynamics. In this paper, we propose a mathematical model for dynamics of cassava mosaic disease that includes immature and mature vectors and explicitly includes a time delay representing vector maturation time. A special feature of our plant epidemic model is that vector recruitment is negatively related to the delayed ratio between vector density and plant density. We identify conditions of biological feasibility and stability of different steady states in terms of system parameters and the time delay. Numerical stability analyses and simulations are performed to explore the role of various parameters, and to illustrate the behaviour of the model in different dynamical regimes. We show that the maturation delay may stabilise epidemiological dynamics that would otherwise be cyclic.


2021 ◽  
Vol 18 (1) ◽  
Author(s):  
Saengsoon Charoenvilaisiri ◽  
Channarong Seepiban ◽  
Mallika Kumpoosiri ◽  
Sombat Rukpratanporn ◽  
Nuchnard Warin ◽  
...  

Abstract Background Cassava mosaic disease (CMD) is one of the most devastating viral diseases for cassava production in Africa and Asia. Accurate yet affordable diagnostics are one of the fundamental tools supporting successful CMD management, especially in developing countries. This study aimed to develop an antibody-based immunoassay for the detection of Sri Lankan cassava mosaic virus (SLCMV), the only cassava mosaic begomovirus currently causing CMD outbreaks in Southeast Asia (SEA). Methods Monoclonal antibodies (MAbs) against the recombinant coat protein of SLCMV were generated using hybridoma technology. MAbs were characterized and used to develop a triple antibody sandwich enzyme-linked immunosorbent assay (TAS-ELISA) for SLCMV detection in cassava leaves and stems. Assay specificity, sensitivity and efficiency for SLCMV detection was investigated and compared to those of a commercial ELISA test kit and PCR, the gold standard. Results A TAS-ELISA for SLCMV detection was successfully developed using the newly established MAb 29B3 and an in-house polyclonal antibody (PAb) against begomoviruses, PAb PK. The assay was able to detect SLCMV in leaves, green bark from cassava stem tips, and young leaf sprouts from stem cuttings of SLCMV-infected cassava plants without cross-reactivity to those derived from healthy cassava controls. Sensitivity comparison using serial dilutions of SLCMV-infected cassava sap extracts revealed that the assay was 256-fold more sensitive than a commercial TAS-ELISA kit and 64-fold less sensitive than PCR using previously published SLCMV-specific primers. In terms of DNA content, our assay demonstrated a limit of detection of 2.21 to 4.08 × 106 virus copies as determined by quantitative real-time PCR (qPCR). When applied to field samples (n = 490), the TAS-ELISA showed high accuracy (99.6%), specificity (100%), and sensitivity (98.2%) relative to the results obtained by the reference PCR. SLCMV infecting chaya (Cnidoscolus aconitifolius) and coral plant (Jatropha multifida) was also reported for the first time in SEA. Conclusions Our findings suggest that the TAS-ELISA for SLCMV detection developed in this study can serve as an attractive tool for efficient, inexpensive and high-throughput detection of SLCMV and can be applied to CMD screening of cassava stem cuttings, large-scale surveillance, and screening for resistance.


1999 ◽  
Vol 354 (1383) ◽  
pp. 521-529 ◽  
Author(s):  
B. D. Harrison ◽  
T. M. A. Wilson

Beijerinck's (1898) recognition that the cause of tobacco mosaic disease was a novel kind of pathogen became the breakthrough which led eventually to the establishment of virology as a science. Research on this agent, tobacco mosaic virus (TMV), has continued to be at the forefront of virology for the past century. After an initial phase, in which numerous biological properties of TMV were discovered, its particles were the first shown to consist of RNA and protein, and X–ray diffraction analysis of their structure was the first of a helical nucleoprotein. In the molecular biological phase of research, TMV RNA was the first plant virus genome to be sequenced completely, its genes were found to be expressed by cotranslational particle disassembly and the use of subgenomic mRNA, and the mechanism of assembly of progeny particles from their separate parts was discovered. Molecular genetical and cell biological techniques were then used to clarify the roles and modes of action of the TMV non–structural proteins: the 126 kDa and 183 kDa replicase components and the 30 kDa cell–to–cell movement protein. Three different TMV genes were found to act as avirulence genes, eliciting hypersensitive responses controlled by specific, but different, plant genes. One of these (the N gene) was the first plant gene controlling virus resistance to be isolated and sequenced. In the biotechnological sphere, TMV has found several applications: as the first source of transgene sequences conferring virus resistance, in vaccines consisting of TMV particles genetically engineered to carry foreign epitopes, and in systems for expressing foreign genes. TMV owes much of its popularity as a research model to the great stability and high yield of its particles. Although modern methods have much decreased the need for such properties, and TMV may have a less dominant role in the future, it continues to occupy a prominent position in both fundamental and applied research.


2004 ◽  
Vol 53 (5) ◽  
pp. 577-584 ◽  
Author(s):  
J. Colvin ◽  
C. A. Omongo ◽  
M. N. Maruthi ◽  
G. W. Otim-Nape ◽  
J. M. Thresh

Viruses ◽  
2021 ◽  
Vol 13 (9) ◽  
pp. 1820
Author(s):  
Warren Freeborough ◽  
Nikki Gentle ◽  
Marie E. C. Rey

Among the numerous biological constraints that hinder cassava (Manihot esculenta Crantz) production, foremost is cassava mosaic disease (CMD) caused by virus members of the family Geminiviridae, genus Begomovirus. The mechanisms of CMD tolerance and susceptibility are not fully understood; however, CMD susceptible T200 and tolerant TME3 cassava landraces have been shown to exhibit different large-scale transcriptional reprogramming in response to South African cassava mosaic virus (SACMV). Recent identification of 85 MeWRKY transcription factors in cassava demonstrated high orthology with those in Arabidopsis, however, little is known about their roles in virus responses in this non-model crop. Significant differences in MeWRKY expression and regulatory networks between the T200 and TME3 landraces were demonstrated. Overall, WRKY expression and associated hormone and enriched biological processes in both landraces reflect oxidative and other biotic stress responses to SACMV. Notably, MeWRKY11 and MeWRKY81 were uniquely up and downregulated at 12 and 67 days post infection (dpi) respectively in TME3, implicating a role in tolerance and symptom recovery. AtWRKY28 and AtWRKY40 homologs of MeWRKY81 and MeWRKY11, respectively, have been shown to be involved in regulation of jasmonic and salicylic acid signaling in Arabidopsis. AtWRKY28 is an interactor in the RPW8-NBS resistance (R) protein network and downregulation of its homolog MeWRKY81 at 67 dpi in TME3 suggests a negative role for this WRKY in SACMV tolerance. In contrast, in T200, nine MeWRKYs were differentially expressed from early (12 dpi), middle (32 dpi) to late (67 dpi) infection. MeWRKY27 (homolog AtWRKY33) and MeWRKY55 (homolog AtWRKY53) were uniquely up-regulated at 12, 32 and 67 dpi in T200. AtWRKY33 and AtWRKY53 are positive regulators of leaf senescence and oxidative stress in Arabidopsis, suggesting MeWRKY55 and 27 contribute to susceptibility in T200.


Plant Disease ◽  
1999 ◽  
Vol 83 (4) ◽  
pp. 398-398 ◽  
Author(s):  
F. O. Ogbe ◽  
G. I. Atiri ◽  
D. Robinson ◽  
S. Winter ◽  
A. G. O. Dixon ◽  
...  

Cassava (Manihot esculenta Crantz) is an important food crop in sub-Saharan Africa. One of the major production constraints is cassava mosaic disease caused by African cassava mosaic (ACMV) and East African cassava mosaic (EACMV) begomoviruses. ACMV is widespread in its distribution, occurring throughout West and Central Africa and in some eastern and southern African countries. In contrast, EACMV has been reported to occur mainly in more easterly areas, particularly in coastal Kenya and Tanzania, Malawi, and Madagascar. In 1997, a survey was conducted in Nigeria to determine the distribution of ACMV and its strains. Samples from 225 cassava plants showing mosaic symptoms were tested with ACMV monoclonal antibodies (MAbs) in triple antibody sandwich enzyme-linked immunosorbent assay (1). Three samples reacted strongly with MAbs that could detect both ACMV and EACMV. One of them did not react with ACMV-specific MAbs while the other two reacted weakly with such MAbs. With polymerase chain reaction (2), the presence of EACMV and a mixture of EACMV and ACMV in the respective samples was confirmed. These samples were collected from two villages: Ogbena in Kwara State and Akamkpa in Cross River State. Co-infection of some cassava varieties with ACMV and EACMV leads to severe symptoms. More importantly, a strain of mosaic geminivirus known as Uganda variant arose from recombination between the two viruses (2). This report provides evidence for the presence of EACMV in West Africa. References: (1) J. E. Thomas et al. J. Gen. Virol. 67:2739, 1986. (2) X. Zhou et al. J. Gen. Virol. 78:2101, 1997.


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