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1.
Phytopathology ; 114(2): 484-495, 2024 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-38408034

RESUMO

Maize lethal necrosis (MLN) is a viral disease caused by host co-infection by maize chlorotic mottle virus (MCMV) and a potyvirus, such as sugarcane mosaic virus (SCMV). The disease is most effectively managed by growing MLN-resistant varieties. However, the relative importance of MCMV and potyvirus resistance in managing this synergistic disease is poorly characterized. In this study, we evaluated the effects of SCMV and/or MCMV resistance on disease, virus titers, and synergism and explored expression patterns of known potyvirus resistance genes TrxH and ABP1. MLN disease was significantly lower in both the MCMV-resistant and SCMV-resistant inbred lines compared with the susceptible control Oh28. Prior to 14 days postinoculation (dpi), MCMV titers in resistant lines N211 and KS23-6 were more than 100,000-fold lower than found in the susceptible Oh28. However, despite no visible symptoms, titer differences between MCMV-resistant and -susceptible lines were negligible by 14 dpi. In contrast, systemic SCMV titers in the potyvirus-resistant line, Pa405, ranged from 130,000-fold to 2 million-fold lower than susceptible Oh28 as disease progressed. Initial TrxH expression was up to 49,000-fold lower in Oh28 compared with other genotypes, whereas expression of ABP1 was up to 4.5-fold lower. Measures of virus synergy indicate that whereas MCMV resistance is effective in early infection, strong potyvirus resistance is critical for reducing synergist effects of co-infection on MCMV titer. These results emphasize the importance of both potyvirus resistance and MCMV resistance in an effective breeding program for MLN management.


Assuntos
Coinfecção , Potyvirus , Tombusviridae , Doenças das Plantas , Necrose
2.
Phytopathology ; 113(9): 1622-1629, 2023 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-37311729

RESUMO

Plant viruses pose a continuous and serious threat to crop production worldwide, and globalization and climate change are exacerbating the establishment and rapid spread of new viruses. Simultaneously, developments in genome sequencing technology, nucleic acid amplification methods, and epidemiological modeling are providing plant health specialists with unprecedented opportunities to confront these major threats to the food security and livelihoods of millions of resource-constrained smallholders. In this perspective, we have used recent examples of integrated application of these technologies to enhance understanding of the emergence of plant viral diseases of key food security crops in low- and middle-income countries. We highlight how international funding and collaboration have enabled high-throughput sequencing-based surveillance approaches, targeted field and lab-based diagnostic tools, and modeling approaches that can be effectively used to support surveillance and preparedness against existing and emerging plant viral threats. The importance of national and international collaboration and the future role of CGIAR in further supporting these efforts, including building capabilities to make optimal use of these technologies in low- and middle-income countries, are discussed. [Formula: see text] Copyright © 2023 The Author(s). This is an open access article distributed under the CC BY 4.0 International license.


Assuntos
Vírus de Plantas , Viroses , Doenças das Plantas , Produtos Agrícolas , Segurança Alimentar
3.
Int J Mol Sci ; 24(9)2023 Apr 28.
Artigo em Inglês | MEDLINE | ID: mdl-37175719

RESUMO

Maize lethal necrosis (MLN), one of the most important maize viral diseases, is caused by maize chlorotic mottle virus (MCMV) infection in combination with a potyvirid, such as sugarcane mosaic virus (SCMV). However, the resistance mechanism of maize to MLN remains largely unknown. In this study, we obtained isoform expression profiles of maize after SCMV and MCMV single and synergistic infection (S + M) via comparative analysis of SMRT- and Illumina-based RNA sequencing. A total of 15,508, 7567, and 2378 differentially expressed isoforms (DEIs) were identified in S + M, MCMV, and SCMV libraries, which were primarily involved in photosynthesis, reactive oxygen species (ROS) scavenging, and some pathways related to disease resistance. The results of virus-induced gene silencing (VIGS) assays revealed that silencing of a vitamin C biosynthesis-related gene, ZmGalDH or ZmAPX1, promoted viral infections, while silencing ZmTAT or ZmNQO1, the gene involved in vitamin E or K biosynthesis, inhibited MCMV and S + M infections, likely by regulating the expressions of pathogenesis-related (PR) genes. Moreover, the relationship between viral infections and expression of the above four genes in ten maize inbred lines was determined. We further demonstrated that the exogenous application of vitamin C could effectively suppress viral infections, while vitamins E and K promoted MCMV infection. These findings provide novel insights into the gene regulatory networks of maize in response to MLN, and the roles of vitamins C, E, and K in conditioning viral infections in maize.


Assuntos
Ácido Ascórbico , Potyvirus , Transcriptoma , Potyvirus/fisiologia , Vitaminas , Zea mays/genética , Doenças das Plantas/genética
4.
Plant Dis ; 105(6): 1596-1601, 2021 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-33320046

RESUMO

Maize chlorotic mottle virus (MCMV) has driven the emergence of maize lethal necrosis worldwide, where it threatens maize production in areas of East Africa, South America, and Asia. It is thought that MCMV transmission through seed may be important for introduction of the virus in new regions. Identification of infested seed lots is critical for preventing the spread of MCMV through seed. Although methods for detecting MCMV in leaf tissue are available, diagnostic methods for its detection in seed lots are lacking. In this study, ELISA, RT-PCR, and RT-qPCR were adapted for detection of MCMV in maize seed. Purified virions of MCMV isolates from Kansas, Mexico, and Kenya were then used to determine the virus detection thresholds for each diagnostic assay. No substantial differences in response were detected among the isolates in any of the three assays. The RT-PCR and a SYBR Green-based RT-qPCR assays were >3,000 times more sensitive than commercial ELISA for MCMV detection. For ELISA using seed extracts, selection of positive and negative controls was critical, most likely because of relatively high backgrounds. Use of seed soak solutions in ELISA detected MCMV with similar sensitivity to seed extracts, produced minimal background, and required substantially less labor. ELISA and RT-PCR were both effective for detecting MCMV in seed lots from Hawaii and Kenya, with ELISA providing a reliable and inexpensive diagnostic assay that could be implemented routinely in seed testing facilities.[Formula: see text] Copyright © 2021 The Author(s). This is an open access article distributed under the CC BY 4.0 International license.


Assuntos
Doenças das Plantas , Tombusviridae , Quênia , Sementes
5.
Plant Dis ; 105(5): 1259-1271, 2021 May.
Artigo em Inglês | MEDLINE | ID: mdl-33289406

RESUMO

Maize lethal necrosis (MLN) disease appeared in Kenya in 2011, causing major damage. In a first survey of 121 communities in 2013, participants estimated the proportion of households affected and the yield loss in affected areas; from this survey, the overall loss was estimated at 22%, concentrated in western Kenya (94%). Efforts to combat the disease included planting resistant varieties, creating awareness of MLN management, and producing pathogen-free seed. In 2018, the same communities were revisited and asked the same questions, establishing a panel community survey. The results showed that incidents of MLN had greatly decreased, and the number of communities that had observed it had reduced from 76% in 2013 to 26% by the long rains of 2018; while still common in western Kenya (60%), MLN had greatly reduced elsewhere (to 10%). In 2013, 40% of farmers were affected, yield loss among affected farmers was estimated at 44%, and total yield loss was estimated at 22% (a production loss of 0.5 million metric tons/year), valued at US$187 million. By the long rains of 2018, 23% of farmers were affected, with a loss among affected farmers of 36%; overall annual loss was estimated at 8.5% or 0.37 million metric tons, valued at US$109 million, concentrated in western Kenya (79%). Of the recommended control measures, only the removal of diseased plants was commonly used (by 62% of affected communities), but not the use of agronomic practices (11%) or resistant varieties (9.5%). The reasons for the reduction in MLN are not well understood; external factors such as spraying insecticide against fall armyworm and unfavorable weather likely played a role, as did using disease-free seed, but not the use of resistant varieties or appropriate management practices. Still, as the pathogen remains in the fields, it is important to keep disseminating these control methods, particularly resistant varieties.[Formula: see text] Copyright © 2021 The Author(s). This is an open access article distributed under the CC BY 4.0 International license.


Assuntos
Sementes , Zea mays , Quênia , Necrose , Inquéritos e Questionários
6.
Virus Genes ; 55(3): 429-432, 2019 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-30790190

RESUMO

Typically associated with fungal species, members of the viral family Totiviridae have recently been shown to be associated with plants, including important crop species, such as Carica papaya (papaya) and Zea mays (maize). Maize-associated totivirus (MATV) was first described in China and more recently in Ecuador, where it has been found to co-occur with other viruses known to elicit maize lethal necrosis disease (MLND). In a survey for maize-associated viruses, 35 samples were selected for Illumina HiSeq sequencing, from the Tanzanian maize producing regions of Mara, Arusha, Manyara, Kilimanjaro, Morogoro and Pwani. Libraries were prepared using an RNA-tag-seq methodology. Taxonomic classification of the resulting datasets showed that 6 of the 35 samples from the regions of Arusha, Kilimanjaro, Morogoro and Mara, contained reads that were assigned to MATV reference sequences. This was confirmed with PCR and Sanger sequencing. Read assembly of the six MATV-associated datasets yielded partial MATV genomes, two of which were selected for further characterization, using RACE. This yielded two full-length MATV genomes, one of which is divergent from other available MATV genomes.


Assuntos
Variação Genética , Doenças das Plantas/virologia , Totivirus/genética , Zea mays/virologia , China , Genoma Viral/genética , Filogenia , Doenças das Plantas/genética , Totivirus/patogenicidade , Zea mays/genética
7.
Virol J ; 15(1): 90, 2018 05 23.
Artigo em Inglês | MEDLINE | ID: mdl-29792207

RESUMO

BACKGROUND: Maize lethal necrosis is caused by a synergistic co-infection of Maize chlorotic mottle virus (MCMV) and a specific member of the Potyviridae, such as Sugarcane mosaic virus (SCMV), Wheat streak mosaic virus (WSMV) or Johnson grass mosaic virus (JGMV). Typical maize lethal necrosis symptoms include severe yellowing and leaf drying from the edges. In Kenya, we detected plants showing typical and atypical symptoms. Both groups of plants often tested negative for SCMV by ELISA. METHODS: We used next-generation sequencing to identify viruses associated to maize lethal necrosis in Kenya through a metagenomics analysis. Symptomatic and asymptomatic leaf samples were collected from maize and sorghum representing sixteen counties. RESULTS: Complete and partial genomes were assembled for MCMV, SCMV, Maize streak virus (MSV) and Maize yellow dwarf virus-RMV (MYDV-RMV). These four viruses (MCMV, SCMV, MSV and MYDV-RMV) were found together in 30 of 68 samples. A geographic analysis showed that these viruses are widely distributed in Kenya. Phylogenetic analyses of nucleotide sequences showed that MCMV, MYDV-RMV and MSV are similar to isolates from East Africa and other parts of the world. Single nucleotide polymorphism, nucleotide and polyprotein sequence alignments identified three genetically distinct groups of SCMV in Kenya. Variation mapped to sequences at the border of NIb and the coat protein. Partial genome sequences were obtained for other four potyviruses and one polerovirus. CONCLUSION: Our results uncover the complexity of the maize lethal necrosis epidemic in Kenya. MCMV, SCMV, MSV and MYDV-RMV are widely distributed and infect both maize and sorghum. SCMV population in Kenya is diverse and consists of numerous strains that are genetically different to isolates from other parts of the world. Several potyviruses, and possibly poleroviruses, are also involved.


Assuntos
Gammaherpesvirinae/genética , Genoma Viral , Luteoviridae/genética , Potyviridae/genética , Potyvirus/genética , Zea mays/virologia , Sequência de Aminoácidos , Proteínas do Capsídeo/genética , Mapeamento Cromossômico , Gammaherpesvirinae/classificação , Gammaherpesvirinae/isolamento & purificação , Gammaherpesvirinae/patogenicidade , Sequenciamento de Nucleotídeos em Larga Escala , Quênia , Luteoviridae/classificação , Luteoviridae/isolamento & purificação , Luteoviridae/patogenicidade , Metagenômica/métodos , Filogenia , Doenças das Plantas/virologia , Folhas de Planta/virologia , Polimorfismo Genético , Potyviridae/classificação , Potyviridae/isolamento & purificação , Potyviridae/patogenicidade , Potyvirus/classificação , Potyvirus/isolamento & purificação , Potyvirus/patogenicidade , Alinhamento de Sequência , Homologia de Sequência de Aminoácidos , Sorghum/virologia
8.
Front Artif Intell ; 7: 1384709, 2024.
Artigo em Inglês | MEDLINE | ID: mdl-39219699

RESUMO

Agriculture is considered the backbone of Tanzania's economy, with more than 60% of the residents depending on it for survival. Maize is the country's dominant and primary food crop, accounting for 45% of all farmland production. However, its productivity is challenged by the limitation to detect maize diseases early enough. Maize streak virus (MSV) and maize lethal necrosis virus (MLN) are common diseases often detected too late by farmers. This has led to the need to develop a method for the early detection of these diseases so that they can be treated on time. This study investigated the potential of developing deep-learning models for the early detection of maize diseases in Tanzania. The regions where data was collected are Arusha, Kilimanjaro, and Manyara. Data was collected through observation by a plant. The study proposed convolutional neural network (CNN) and vision transformer (ViT) models. Four classes of imagery data were used to train both models: MLN, Healthy, MSV, and WRONG. The results revealed that the ViT model surpassed the CNN model, with 93.1 and 90.96% accuracies, respectively. Further studies should focus on mobile app development and deployment of the model with greater precision for early detection of the diseases mentioned above in real life.

9.
Data Brief ; 48: 109108, 2023 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-37095756

RESUMO

Maize is one of the most important staple food and cash crops that are largely produced by majority of smallholder farmers throughout the humid and sub-humid tropic of Africa. Despite its significance in the household food security and income, diseases, especially Maize Lethal Necrosis and Maize Streak, have been significantly affecting production of this crop. This paper offers a dataset of well curated images of maize crop for both healthy and diseased leaves captured using smartphone camera in Tanzania. The dataset is the largest publicly accessible dataset for maize leaves with a total of 18,148 images, which can be used to develop machine learning models for the early detection of diseases affecting maize. Moreover, the dataset can be used to support computer vision applications such as image segmentation, object detection and classification. The goal of generating this dataset is to assist the development of comprehensive tools that will help farmers in the diagnosis of diseases and the enhancement of maize yields thus eradicating the problem of fod security in Tanzania and other parts in Africa.

10.
PeerJ ; 11: e15685, 2023.
Artigo em Inglês | MEDLINE | ID: mdl-38050609

RESUMO

Maize (Zea mays L.) is a staple food for many households in sub-Saharan Africa (SSA) and also contributes to the gross domestic product (GDP). However, the maize yields reported in most SSA countries are very low and this is mainly attributed to biotic and abiotic stresses. These stresses have been exacerbated by climate change which has led to long periods of drought or heavy flooding and the emergence of new biotic stresses. Few reports exist which compile the biotic stresses affecting maize production in SSA. Here, five major biotic stresses of maize in Kenya are presented which are attributed to high yield losses. They include Maize lethal necrosis, fall armyworm, gray leaf spot, turcicum leaf blight and desert locusts. Maize lethal necrosis and fall armyworm are new biotic stresses to the Kenyan maize farmer while gray leaf spot, and turcicum leaf blight are endemic to the region. The invasion by the desert locusts is speculated to be caused by climate change. The biotic stresses cause a reduction in maize yield of 30-100% threatening food security. Therefore, this review focuses on the cause, control measures employed to control these diseases and future prospective. There should be deliberate efforts from the government and researchers to control biotic stresses affecting maize yields as the effect of these stresses is being exacerbated by the changing climate.


Assuntos
Doenças das Plantas , Zea mays , Quênia , Estresse Fisiológico , Segurança Alimentar , Necrose
11.
Viruses ; 14(12)2022 12 12.
Artigo em Inglês | MEDLINE | ID: mdl-36560769

RESUMO

Maize Lethal Necrosis (MLN) disease, caused by a synergistic co-infection of maize chlorotic mottle virus (MCMV) and any member of the Potyviridae family, was first reported in EasternAfrica (EA) a decade ago. It is one of the most devastating threats to maize production in these regions since it can lead up to 100% crop loss. Conventional counter-measures have yielded some success; however, they are becoming less effective in controlling MLN. In EA, the focus has been on the screening and identification of resistant germplasm, dissecting genetic and the molecular basis of the disease resistance, as well as employing modern breeding technologies to develop novel varieties with improved resistance. CIMMYT and scientists from NARS partner organizations have made tremendous progresses in the screening and identification of the MLN-resistant germplasm. Quantitative trait loci mapping and genome-wide association studies using diverse, yet large, populations and lines were conducted. These remarkable efforts have yielded notable outcomes, such as the successful identification of elite resistant donor lines KS23-5 and KS23-6 and their use in breeding, as well as the identification of multiple MLN-tolerance promising loci clustering on Chr 3 and Chr 6. Furthermore, with marker-assisted selection and genomic selection, the above-identified germplasms and loci have been incorporated into elite maize lines in a maize breeding program, thus generating novel varieties with improved MLN resistance levels. However, the underlying molecular mechanisms for MLN resistance require further elucidation. Due to third generation sequencing technologies as well functional genomics tools such as genome-editing and DH technology, it is expected that the breeding time for MLN resistance in farmer-preferred maize varieties in EA will be efficient and shortened.


Assuntos
Estudo de Associação Genômica Ampla , Potyviridae , Resistência à Doença/genética , Doenças das Plantas/genética , Locos de Características Quantitativas , Potyviridae/genética , Zea mays/genética , Fenótipo
12.
Genes (Basel) ; 13(2)2022 02 15.
Artigo em Inglês | MEDLINE | ID: mdl-35205395

RESUMO

Breeding maize lines with the improved level of desired agronomic traits under optimum and drought conditions as well as increased levels of resistance to several diseases such as maize lethal necrosis (MLN) is one of the most sustainable approaches for the sub-Saharan African region. In this study, 879 doubled haploid (DH) lines derived from 26 biparental populations were evaluated under artificial inoculation of MLN, as well as under well-watered (WW) and water-stressed (WS) conditions for grain yield and other agronomic traits. All DH lines were used for analyses of genotypic variability, association studies, and genomic predictions for the grain yield and other yield-related traits. Genome-wide association study (GWAS) using a mixed linear FarmCPU model identified SNPs associated with the studied traits i.e., about seven and eight SNPs for the grain yield; 16 and 12 for anthesis date; seven and eight for anthesis silking interval; 14 and 5 for both ear and plant height; and 15 and 5 for moisture under both WW and WS environments, respectively. Similarly, about 13 and 11 SNPs associated with gray leaf spot and turcicum leaf blight were identified. Eleven SNPs associated with senescence under WS management that had depicted drought-stress-tolerant QTLs were identified. Under MLN artificial inoculation, a total of 12 and 10 SNPs associated with MLN disease severity and AUDPC traits, respectively, were identified. Genomic prediction under WW, WS, and MLN disease artificial inoculation revealed moderate-to-high prediction accuracy. The findings of this study provide useful information on understanding the genetic basis for the MLN resistance, grain yield, and other agronomic traits under MLN artificial inoculation, WW, and WS conditions. Therefore, the obtained information can be used for further validation and developing functional molecular markers for marker-assisted selection and for implementing genomic prediction to develop superior elite lines.


Assuntos
Resistência à Doença , Estudo de Associação Genômica Ampla , Resistência à Doença/genética , Grão Comestível/genética , Haploidia , Fenótipo , Melhoramento Vegetal , Zea mays/genética
13.
Front Plant Sci ; 13: 889995, 2022.
Artigo em Inglês | MEDLINE | ID: mdl-35712601

RESUMO

Recent advances in biotechnology have helped increase tissue transformation efficiency and the frequency and specificity of gene editing to an extent that introducing allelic variants directly in elite varieties has become possible. In comparison to the conventional approach of crossing an elite recipient line with an exotic donor parent to introduce the trait of interest followed by repeated backcrossing, direct introduction of major-effect allelic variants into elite varieties saves time and resources, and eliminates yield drag resulting from the residual donor genes at the end of backcrossing.

14.
Front Plant Sci ; 12: 649308, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-34040620

RESUMO

Maize lethal necrosis (MLN), resulting from co-infection by maize chlorotic mottle virus (MCMV) and sugarcane mosaic virus (SCMV) can cause up to 100% yield losses in maize in Africa under serious disease conditions. Maize improvement through conventional backcross (BC) takes many generations but can significantly be shortened when molecular tools are utilized in the breeding process. We used a donor parent (KS23-6) to transfer quantitative trait loci (QTL) for resistance to MLN into nine adapted but MLN susceptible lines. Nurseries were established in Kiboko, Kenya during 2015-2017 seasons and BC3F2 progeny were developed using marker assisted backcrossing (MABC) approach. Six single nucleotide polymorphism (SNP) markers linked to QTL for resistance to MLN were used to genotype 2,400 BC3F2 lines using Kompetitive Allele Specific PCR (KASP) platform. We detected that two of the six QTL had major effects for resistance to MLN under artificial inoculation field conditions in 56 candidate BC3F2 lines. To confirm whether these two QTL are reproducible under different field conditions, the 56 BC3F2 lines including their parents were evaluated in replicated trials for two seasons under artificial MLN inoculations in Naivasha, Kenya in 2018. Strong association of genotype with phenotype was detected. Consequently, 19 superior BC3F2 lines with favorable alleles and showing improved levels of resistance to MLN under artificial field inoculation were identified. These elite lines represent superior genetic resources for improvement of maize hybrids for resistance to MLN. However, 20 BC3F2 lines were fixed for both KASP markers but were susceptible to MLN under field conditions, which could suggest weak linkage between the KASP markers and target genes. The validated two major QTL can be utilized to speed up the breeding process but additional loci need to be identified between the KASP markers and the resistance genes to strengthen the linkage.

15.
Front Genet ; 12: 767883, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-34868253

RESUMO

Maize lethal necrosis (MLN) is a viral disease with a devastating effect on maize production. Developing and deploying improved varieties with resistance to the disease is important to effectively control MLN; however, little is known about the causal genes and molecular mechanism(s) underlying MLN resistance. Screening thousands of maize inbred lines revealed KS23-5 and KS23-6 as two of the most promising donors of MLN resistance alleles. KS23-5 and KS23-6 lines were earlier developed at the University of Hawaii, United States, on the basis of a source population constituted using germplasm from Kasetsart University, Thailand. Both linkage mapping and association mapping approaches were used to discover and validate genomic regions associated with MLN resistance. Selective genotyping of resistant and susceptible individuals within large F2 populations coupled with genome-wide association study identified a major-effect QTL (qMLN06_157) on chromosome 6 for MLN disease severity score and area under the disease progress curve values in all three F2 populations involving one of the KS23 lines as a parent. The major-effect QTL (qMLN06_157) is recessively inherited and explained 55%-70% of the phenotypic variation with an approximately 6 Mb confidence interval. Linkage mapping in three F3 populations and three F2 populations involving KS23-5 or KS23-6 as one of the parents confirmed the presence of this major-effect QTL on chromosome 6, demonstrating the efficacy of the KS23 allele at qMLN06.157 in varying populations. This QTL could not be identified in population that was not derived using KS23 lines. Validation of this QTL in six F2 populations with 20 SNPs closely linked with qMLN06.157 was further confirmed its consistent expression across populations and its recessive nature of inheritance. On the basis of the consistent and effective resistance afforded by the KS23 allele at qMLN06.157, the QTL can be used in both marker-assisted forward breeding and marker-assisted backcrossing schemes to improve MLN resistance of breeding populations and key lines for eastern Africa.

16.
J Virol Methods ; 298: 114286, 2021 12.
Artigo em Inglês | MEDLINE | ID: mdl-34520808

RESUMO

Maize lethal necrosis disease (MLND) is a serious disease of worldwide importance. It is caused by the co-infection of maize with maize chlorotic mottle virus (MCMV) and a potyvirus, such as sugarcane mosaic virus (SCMV), that acts synergistically to produce more severe symptoms and production losses. More recently, maize yellow mosaic virus (MaYMV) and maize-associated totivirus (MATV) were found to co-infect with MCMV and SCMV in maize plants. To facilitate the detection of these viruses in co-infected maize, a multiplex RT-PCR assay was developed in this study. The assay used five specific primer pairs and simultaneously amplified these four viruses as well as the elongation factor 1α (EF 1α) gene use as internal control in one tube. The concentration of the primers, annealing temperature, annealing time, extension time and amplification cycles were optimized for the multiplex RT-PCR. The detection limit of the assay was up to 100 pg of total cDNA template. This multiplex RT-PCR assay was shown to be a sensitive and effective tool for the screening of field samples for the presence of these viruses in co-infected maize.


Assuntos
Luteoviridae , Doenças das Plantas , Humanos , Luteoviridae/genética , Necrose , Reação em Cadeia da Polimerase Via Transcriptase Reversa
17.
Plants (Basel) ; 9(4)2020 Apr 08.
Artigo em Inglês | MEDLINE | ID: mdl-32276322

RESUMO

Prior knowledge on heterosis and quantitative genetic parameters on maize lethal necrosis (MLN) can help the breeders to develop numerous resistant or tolerant hybrids with optimum resources. Our objectives were to (1) estimate the quantitative genetic parameters for MLN disease severity, (2) investigate the efficiency of the prediction of hybrid performance based on parental per se and general combining ability (GCA) effects, and (3) examine the potential of hybrid prediction for MLN resistance or tolerance based on markers. Fifty elite maize inbred lines were selected based on their response to MLN under artificial inoculation. Crosses were made in a half diallel mating design to produce 307 F1 hybrids. All hybrids were evaluated in MLN quarantine facility in Naivasha, Kenya for two seasons under artificial inoculation. All 50 inbreds were genotyped with genotyping-by-sequencing (GBS) SNPs. The phenotypic variation was significant for all traits and the heritability was moderate to high. We observed that hybrids were superior to the mean performance of the parents for disease severity (-14.57%) and area under disease progress curve (AUDPC) (14.9%). Correlations were significant and moderate between line per se and GCA; and mean of parental value with hybrid performance for both disease severity and AUDPC value. Very low and negative correlation was observed between parental lines marker based genetic distance and heterosis. Nevertheless, the correlation of GCA effects was very high with hybrid performance which can suggests as a good predictor of MLN resistance. Genomic prediction of hybrid performance for MLN is high for both traits. We therefore conclude that there is potential for prediction of hybrid performance for MLN. Overall, the estimated quantitative genetic parameters suggest that through targeted approach, it is possible to develop outstanding lines and hybrids for MLN resistance.

18.
Plant Pathol ; 69(3): 585-597, 2020 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-35874461

RESUMO

Maize lethal necrosis (MLN) is emergent in East Africa, first reported in 2011 in Kenya, and is devastating to maize production in the region. MLN is caused by coinfection of maize with the emergent maize chlorotic mottle virus (MCMV) and any of several maize-infecting potyviruses endemic in East Africa and worldwide. Here, we examined the distribution of MCMV and sugarcane mosaic virus (SCMV), the major viruses contributing to MLN in Rwanda. These and other viruses in maize across Rwanda were further characterized by deep sequencing. When identified, MCMV had high titres and minimal sequence variability, whereas SCMV showed moderate titres and high sequence variability. Deep sequencing also identified maize streak virus and other maize-associated viruses, including a previously described polerovirus, maize yellow mosaic virus, and barley yellow dwarf virus, diverse maize-associated totiviruses, maize-associated pteridovirus, Zea mays chrysovirus 1, and a maize-associated betaflexivirus. Detection of each virus was confirmed in maize samples by reverse transcription polymerase chain reaction.

19.
Plants (Basel) ; 9(4)2020 Mar 28.
Artigo em Inglês | MEDLINE | ID: mdl-32231080

RESUMO

In eastern Africa, Maize lethal necrosis (MLN) is caused by the co-infection of maize plants with Maize chlorotic mottle virus (MCMV) (Tombusviridae: Machlomovirus) and Sugarcane mosaic virus (SCMV) (Potyviridae: Potyvirus). With the disease being new to Africa, minimal effective management strategies exist against it. This study examined the potential of 10 fungal isolates to colonize maize plants and induce resistance against MCMV and SCMV. Maize seeds were soaked in fungal inoculum, sown and evaluated for endophytic colonization. Fungus-treated plants were challenge-inoculated with SCMV and/or MCMV to assess the effects of fungal isolates on the viruses in terms of incidence, severity and virus titers over time. Isolates of Trichoderma harzianum, Trichoderma atroviride and Hypocrea lixii colonized different plant sections. All plants singly or dually-inoculated with SCMV and MCMV tested positive for the viruses by reverse transcription-polymerase chain reaction (RT-PCR). Maize plants inoculated by T. harzianum and Metarhizium. anisopliae resulted in up to 1.4 and 2.7-fold reduced SCMV severity and titer levels, respectively, over the controls but had no significant effect on MCMV. The results show that both T. harzianum and M. anisopliae are potential candidates for inducing resistance against SCMV and can be used for the integrated management of MLN.

20.
Virus Res ; 282: 197943, 2020 06.
Artigo em Inglês | MEDLINE | ID: mdl-32205142

RESUMO

Maize lethal necrosis (MLN), a complex viral disease, emerged as a serious threat to maize production and the livelihoods of smallholders in eastern Africa since 2011, primarily due to the introduction of maize chlorotic mottle virus (MCMV). The International Maize and Wheat Improvement Center (CIMMYT), in close partnership with national and international partners, implemented a multi-disciplinary and multi-institutional strategy to curb the spread of MLN in sub-Saharan Africa, and mitigate the impact of the disease. The strategy revolved around a) intensive germplasm screening and fast-tracked development and deployment of MLN-tolerant/resistant maize hybrids in Africa-adapted genetic backgrounds; b) optimizing the diagnostic protocols for MLN-causing viruses, especially MCMV, and capacity building of relevant public and private sector institutions on MLN diagnostics and management; c) MLN monitoring and surveillance across sub-Saharan Africa in collaboration with national plant protection organizations (NPPOs); d) partnership with the private seed sector for production and exchange of MLN pathogen-free commercial maize seed; and e) awareness creation among relevant stakeholders about MLN management, including engagement with policy makers. The review concludes by highlighting the need to keep continuous vigil against MLN-causing viruses, and preventing any further spread of the disease to the major maize-growing countries that have not yet reported MLN in sub-Saharan Africa.


Assuntos
Doenças das Plantas/prevenção & controle , Doenças das Plantas/virologia , Tombusviridae/patogenicidade , Zea mays/virologia , África Subsaariana , Necrose , Sementes/virologia
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