Protocols for the Detection and Proteome Analysis of the Yellow Mosaic Virus Infected Soyabean Leaves

Soybean (Glycine max) is one of the legumes, susceptible to yellow mosaic disease caused by Mungbean yellow mosaic India virus (MYMIV) and Mungbean yellow mosaic virus (MYMV) infection. The quantitative proteomic analysis allows achieving deeper knowledge about the viral infection. For quantitative proteomic analysis, two-dimensional gel electrophoresis (2D-PAGE) is the common method of choice. Optimization is required even for the published protocols based on the type of sample to be analyzed and for the proteins of interest. We compared four different published protocols with some modifications and selected the one which is more effective in terms of resolution and reproducibility of 2D-PAGE. Here we present our simple and cost-effective procedure for the detection of viral infection and proteomic analysis of YMV infected soybean leaves without compromising the resolution and reproducibility of 2D-PAGE.

Agronomy ◽  
2019 ◽  
Vol 9 (10) ◽  
pp. 622 ◽  
Author(s):  
Chandra Mohan Singh ◽  
Poornima Singh ◽  
Aditya Pratap ◽  
Rakesh Pandey ◽  
Shalini Purwar ◽  
...  

Yellow mosaic disease (YMD) affects several types of leguminous crops, including the Vigna species, which comprises a number of commercially important pulse crops. YMD is characterized by the formation of a bright yellow mosaic pattern on the leaves; in severe forms, this pattern can also be seen on stems and pods. This disease leads to tremendous yield losses, even up to 100%, in addition to deterioration in seed quality. Symptoms of this disease are similar among affected plants; YMD is not limited to mungbean (Vigna radiata L. Wilczek) and also affects other collateral and alternate hosts. In the last decade, rapid advancements in molecular detection techniques have been made, leading to an improved understanding of YMD-causing viruses. Three distinct bipartite begomoviruses, namely, Mungbean Yellow Mosaic India Virus (MYMIV), Mungbean Yellow Mosaic Virus (MYMV), and Horsegram Yellow Mosaic Virus (HgYMV), are known to cause YMD in Vigna spp. Vigna crops serve as an excellent protein source for vegetarians worldwide; moreover, they aid in improving soil health by fixing atmospheric nitrogen through a symbiotic association with Rhizobium bacteria. The loss in the yield of these short-duration crops due to YMD, thus, needs to be checked. This review highlights the discoveries that have been made regarding various aspects of YMD affecting mungbean, including the determination of YMD-causing viruses and strategies used to develop high-yielding YMD-resistant mungbean varieties that harness the potential of related Vigna species through the use of different omics approaches.


Author(s):  
Sudeep Pandey ◽  
T.R. Girish ◽  
S. Basavaraj ◽  
A.S. Padmaja ◽  
N. Nagaraju

Background: Yellow mosaic disease (YMD) caused by begomoviruses transmitted through the insect vector Bemisia tabaci poses a serious threat to the production of legume crops. Methods: Season-long surveys were carried out for YMD occurrence in six different legume crops and associated natural weeds both symptomatic and asymptomatic across the districts of southern Karnataka, India. The samples were analyzed through RCA PCR using specific primer pairs. Result: Up to 94.1 per cent YMD incidence was recorded and nine weed species were commonly found associated with legume crops. The weeds viz., Ageratum conyzoides, Alternanthera sessilis, Commelina benghalensis and Euphorbia geniculata were abundantly found in the surveyed regions. The weeds were both symptomatic and asymptomatic. Rolling circle amplification coupled polymerase chain reaction method was employed to detect yellow mosaic virus in asymptomatic weeds. Phylogenetic analysis based on the sequences of PCR amplified products of weeds and symptomatic legumes revealed a close clustering of the weed samples with horsegram yellow mosaic virus, legume yellow mosaic virus and mungbean yellow mosaic virus. Overall, our data suggests the role of weed species associated with legume crops as alternative/collateral hosts of begomoviruses and their role in the epidemiology of yellow mosaic disease.


Author(s):  
Hélène Pidon ◽  
Neele Wendler ◽  
Antje Habekuβ ◽  
Anja Maasberg ◽  
Brigitte Ruge-Wehling ◽  
...  

Abstract Key message We mapped the Rym14Hb resistance locus to barley yellow mosaic disease in a 2Mbp interval. The co-segregating markers will be instrumental for marker-assisted selection in barley breeding. Abstract Barley yellow mosaic disease is caused by Barley yellow mosaic virus and Barley mild mosaic virus and leads to severe yield losses in barley (Hordeum vulgare) in Central Europe and East-Asia. Several resistance loci are used in barley breeding. However, cases of resistance-breaking viral strains are known, raising concerns about the durability of those genes. Rym14Hb is a dominant major resistance gene on chromosome 6HS, originating from barley’s secondary genepool wild relative Hordeum bulbosum. As such, the resistance mechanism may represent a case of non-host resistance, which could enhance its durability. A susceptible barley variety and a resistant H. bulbosum introgression line were crossed to produce a large F2 mapping population (n = 7500), to compensate for a ten-fold reduction in recombination rate compared to intraspecific barley crosses. After high-throughput genotyping, the Rym14Hb locus was assigned to a 2Mbp telomeric interval on chromosome 6HS. The co-segregating markers developed in this study can be used for marker-assisted introgression of this locus into barley elite germplasm with a minimum of linkage drag.


2018 ◽  
Vol 1 (2) ◽  
pp. p100
Author(s):  
Md. S. Islam ◽  
Md. B. Hossain ◽  
Saleh A. Shahriar ◽  
Fatema Begum ◽  
Md. N. H. Sani

The prime aim of the study was to manage of Yellow mosaic disease of mungbean against Mungbean yellow mosaic virus (MYMV) by using one newly release botanical nutrient and through three selected insecticides. BARI (Bangladesh Agricultural Research Institute) released variety BARI mung-5, three insecticides (Imidacloprid, Acmix and Sobicron) and one botanical nutrient PPN (Peak performance nutrients) were used in the experiment. The plants were grown for pulse production and natural inoculums were relied upon for the infection of MYMV. Growth parameters, yield attributes and physiological features were significantly influenced by the application of selected insecticides and PPN combinations. Disease incidence and disease severity of MYMV were significantly varied among the treatments. Application of Imidacloprid with PPN combination gave the lowest disease incidence (3.13, 5.24 and 6.24% per plot and 14.33, 15.49 and 21.87% per plant) at 30, 40 and 50 DAS, respectively while the highest disease incidence (7.77, 13.70 and 19.24% per plot and 39.33, 48.20 and 56.63% per plant) were found in control at 30, 40 and 50 DAS, respectively. Application of Imidacloprid with PPN also gave the lowest disease severity (5.00, 6.00 and 13.33% at 30, 40 and 50 DAS, respectively while the highest disease severity (27.33, 35.00 and 45.00%) at 30, 40 and 50 DAS, respectively were measured in control treatment when no insecticides and PPN was used. If the disease is established once in the field then it is difficult to manage. As the disease is transmitted by vector (whitefly), the growers are suggested to control the vector populations before reaching economic damage and severe disease infection.


Author(s):  
K. Kamesh Krishnamoorthy ◽  
V. G. Malathi ◽  
P. Renukadevi ◽  
S. Mohan Kumar ◽  
M. Raveendran ◽  
...  

The yellow mosaic disease of blackgram caused by Mungbean yellow mosaic virus has emerged as a serious threat to pulses production especially in the South Eastern Asia.  Seed borne nature of MYMV in blackgram seeds was determined using the seeds harvested from a MYMV resistant (either VBN-6 or VBN-8) and susceptible blackgram (CO-5) varieties grown in three different agroclimatic zones of Tamil Nadu in India for three consecutive cropping seasons namely, Rabi 2018 (October- December), Summer 2019 (March-May) and Kharif 2019 (June- August) at three different time intervals viz., 20, 40 and 60 days after sowing (DAS). Seed borne nature of MYMV was observed only in the susceptible variety CO-5 and was absent in the   resistant varieties. Transmission of MYMV from infected plant to seeds was observed in all the three parts of the seeds viz., seed coat, cotyledon and embryo. Seeds from infected plants also showed abnormalities like shrinking, discolouration, ill filling inside pods and misshapen appearance.


2019 ◽  
Vol 17 (04) ◽  
pp. 362-370
Author(s):  
Nagaraj ◽  
S Basavaraj ◽  
A.S. Padmaja ◽  
N Nagaraju ◽  
S Ramesh

AbstractYellow mosaic disease (YMD) caused by mungbean yellow mosaic virus (MYMV) is one of the most destructive biotic production constraints in mungbean. Development and introduction of resistant cultivars are considered as the most economical and eco-friendly option to manage YMD, for which availability of stable sources of resistance is a pre-requisite. A set of 14 mungbean genotypes including a susceptible check were evaluated for responses to YMD under natural infection across three seasons and under challenged inoculation in glasshouse for one season. None of the genotypes were immune to YMD and produced different degrees of response to MYMV in terms of yellow mosaic symptoms (YMS). Based on the delayed appearance of initial YMS, and lower estimates of per cent disease index and area under disease progressive curve (AUDPC) in response to natural infection and challenged inoculation, five genotypes namely AVMU 1698, AVMU 1699, AVMU 16100, AVMU 16101 and KPS 2 were identified as resistant to YMD. Failure of detection of MYMV through polymerase chain reaction (PCR) using MYMV coat protein gene-specific primer and successful detection of the same through rolling circle amplification-PCR suggested latent infection of MYMV in resistant genotypes. The resistance response of the five genotypes could be attributed to enhanced activities of enzymes such as peroxidase, polyphenol oxidase and phenylalanine ammonia lyase and increased concentration of total phenols. These results are discussed in relation to strategies to breed mungbean for resistance to YMD.


Plant Disease ◽  
2011 ◽  
Vol 95 (8) ◽  
pp. 901-906 ◽  
Author(s):  
J. Karkashian ◽  
E. D. Ramos-Reynoso ◽  
D. P. Maxwell ◽  
P. Ramírez

Begomovirus spp. cause substantial losses in bean crops in tropical and subtropical regions of the Americas. The predominant Begomovirus sp. in Central America associated with golden mosaic symptoms in bean is Bean golden yellow mosaic virus (BGYMV). However, Calopogonium golden mosaic virus was previously found to infect bean crops in the northern region of Costa Rica. The objective of this research was to identify Begomovirus spp. that infect bean plants in different geographical regions of Nicaragua. In all, 126 samples of young bean leaves with symptoms of golden mosaic were collected from eight different regions of Nicaragua. Using DNA hybridization with specific probes, 120 samples tested positive for BGYMV, 14 samples tested positive for Squash yellow mild mottle virus, and 7 samples tested positive for Calopogonium golden mosaic virus. Sequence analysis of polymerase chain reaction-amplified products from three samples (MA-9 Managua, BE-8 Rivas, and SO-9 Granada) also indicated that the symptoms of golden mosaic in bean are associated with viral sequences from three different Begomovirus spp. Management of bean golden mosaic disease must take into account that BGYMV is the predominant virus (95% of the samples) and that 12% of the samples exhibited possible mixed infections or recombination events in the south and central geographical regions of Nicaragua.


2003 ◽  
Vol 77 (17) ◽  
pp. 9124-9135 ◽  
Author(s):  
Delphine Prod'homme ◽  
Anna Jakubiec ◽  
Vincent Tournier ◽  
Gabrièle Drugeon ◽  
Isabelle Jupin

ABSTRACT Turnip yellow mosaic virus (TYMV), a positive-strand RNA virus in the alphavirus-like superfamily, encodes two replication proteins, 140K and 66K, both being required for its RNA genome replication. The 140K protein contains domains indicative of methyltransferase, proteinase, and NTPase/helicase, and the 66K protein encompasses the RNA-dependent RNA polymerase domain. During viral infection, the 66K protein localizes to virus-induced chloroplastic membrane vesicles, which are closely associated with TYMV RNA replication. To investigate the determinants of its subcellular localization, the 66K protein was expressed in plant protoplasts from separate plasmids. Green fluorescent protein (GFP) fusion and immunofluorescence experiments demonstrated that the 66K protein displayed a cytoplasmic distribution when expressed individually but that it was relocated to the chloroplast periphery under conditions in which viral replication occurred. The 66K protein produced from an expression vector was functional in viral replication since it could transcomplement a defective replication template. Targeting of the 66K protein to the chloroplast envelope in the course of the viral infection appeared to be solely dependent on the expression of the 140K protein. Analysis of the subcellular localization of the 140K protein fused to GFP demonstrated that it is targeted to the chloroplast envelope in the absence of other viral factors and that it induces the clumping of the chloroplasts, one of the typical cytological effects of TYMV infection. These results suggests that the 140K protein is a key organizer of the assembly of the TYMV replication complexes and a major determinant for their chloroplastic localization and retention.


Sign in / Sign up

Export Citation Format

Share Document