Chickpea Isolates Research Articles

Effect of cellulose nanofibrils on stability and digestive properties of legume protein-based emulsions

The global sustainability and health consciousness awakening have contributed to the rise of legume proteins, but the poor emulsion stability of legume proteins limits their application. Herein, cellulose nanofibrils (CNFs) isolated from citrus peels were used to improve the stability of pea isolate protein, chickpea isolate protein, and soybean isolate protein emulsions. Results showed that CNFs extracted from citrus peels could improve the interfacial properties of legume proteins via non-covalent interaction. The addition of CNFs could increase interfacial tension and contact angle compared to the legume protein alone. CNFs resulted in the formation of homogeneous emulsions with smaller particle sizes as well as an increase in the viscosity of the emulsions. Meanwhile, the stability of legume protein/CNFs complex emulsions showed significant advantages over single legume protein emulsions in terms of long-term storage, centrifugation, salt ions, and high temperature. Furthermore, legume protein/CNFs emulsions have a lower rate of lipid digestion. These results revealed that CNFs have promising potential for improving legume protein emulsion stability and retarding lipolysis, which provides a theoretical basis for advancing the use of legume proteins.

Levels and types of microbial contaminants in different plant-based ingredients used in dairy alternatives

In this study levels and types of microbial contaminants were investigated in 88 different plant-based ingredients including many that are used to manufacture dairy alternatives. Studied ingredients encompassed samples of pulses (pea, faba bean, chickpea, and mung bean), cereals/pseudocereals (oat, rice, amaranth and quinoa) and drupes (coconut, almond and cashew). The microbial analysis included: i) total viable count (TVC), ii) total aerobic mesophilic spore count (TMS), iii) heat resistant aerobic thermophilic spore count (HRTS), iv) anaerobic sulfite reducing Clostridium spore count (SRCS), and v) Bacillus cereus spore count (BCES). Microorganisms isolated from the counting plates with the highest sample dilutions were identified using 16S rRNA and MALDI-TOF MS analyses.Many of the investigated ingredients showed a high proportion of spores as part of their total aerobic mesophilic counts. In 63 % of the samples, the difference between TVC and TMS counts was 1 Log10 unit or less. This was particularly the case for the majority of pea isolates and concentrates, faba bean isolates, oat kernels and flakes, and for single samples of chickpea isolate, almond, amaranth, rice, quinoa, and coconut flours. Concentrations of TVC ranged between <1.0 and 5.3 Log10 CFU/g in different samples, and TMS varied between <1.0 and 4.1 Log10 CFU/g. Levels of HTRS, BCES and SRCS were generally low, typically around or below the LOD of 1.0 Log10 CFU/g.In total, 845 individual bacterial colonies were isolated belonging to 33 different genera. Bacillus licheniformis and B. cereus group strains were most frequently detected among Bacillus isolates, and these species originated primarily from pea and oat samples. Geobacillus stearothermophilus was the main species encountered as part of the HRTS. Among the Clostridium isolates, Clostridum sporogenes/tepidum were predominant species, which were mostly found in pea and almond samples. Strains with potential to cause foodborne infection or intoxication were typed using the PCR-based method for toxin genes detection. In the B. cereus group, 9 % of isolates contained the ces gene, 28 % contained hbl, 42 % cytK, and 69 % were positive for the nhe gene. Absence of the boNT-A and -B genes was confirmed for all isolated C. sporogenes/tepidum strains. Nearly all (98 %) B. licheniformis isolates were positive for the lchAA gene.Insight into the occurrence of microbial contaminants in plant-based ingredients, combined with knowledge of their key inactivation and growth characteristics, can be used for the microbial risk assessment and effective design of plant-based food processing conditions and formulations to ensure food safety and prevent spoilage.

Utilizing Chickpea Isolates as a Fortificant to Develop Protein-Dense Yogurt: A Review

Pulses and dairy products are recognized for their nutritional and functional benefits and are consumed in various forms. Yogurt is considered a source of good quality protein with anti-carcinogenic, hypocholesterolemic properties, and palliating effects on lactose intolerance. Similarly, chickpea is known for its high protein content, low glycemic index, and hypoglycaemic effects. These food ingredients cater to numerous advantages for human health and can address public health issues related to malnutrition or other nutritional deficiencies. With this background, the manuscript explores the possibility of employing chickpea isolates to fortify yogurt to improvise protein content along with sensory and physicochemical properties. So far, the literature has shown that protein extracts, when added to yogurt, result in gaining protein content and overall product quality. The yogurt market is growing, and consumers from different countries have expressed their willingness to purchase fortified yogurts to achieve optimum health. Therefore, developing a new combination of yogurt and chickpea isolates can provide a therapeutic alternative to enhance the nutritional status of the vulnerable population, viz. children, pregnant, lactating mothers, elderly, sportsperson, etc., when a judicious food intake is a must.

Effect of low-frequency high-intensity ultrasound (HIU) on the physicochemical properties of chickpea protein

The aim of this study was to improve the functional properties of chickpea protein for its potential application in the food industry. The effects of low frequency high intensity ultrasound (HIU) at different power (0–300 W) and time (15–30 min) on the rheological properties, gelation, thermal stability, solubility and microstructure of chickpea protein were tested and analyzed. Based on the analysis, it was found that HIU caused the disruption of non-covalent bonds between protein chains leading to the unfolding of chickpea. The HIU-treated chickpea isolate protein aggregates were smaller and more uniformly dispersed, with increased orderly structure, thermal stability, and exposure of hydrophobic and charged groups originally buried in the interior. The experimental results also showed that the effect of HIU did not become more pronounced with increasing power and time, as the power exceeding 150 W for 30 min led to the formation of new polymers by the interactions between the exposed non-covalent groups, which were more ordered and homogeneous than those without HIU.

Morphological, molecular and pathogenic characterization of Rhizoctonia bataticola isolates causing dry root rot of chickpea in Turkey

Nineteen chickpea isolates of R. bataticola were obtained from different locations of Turkey in 2016–2017. The isolates were examined for their morphology, genetic and virulence characteristics. The colony color of 10 isolates was grayish black and rest of them was black. Eleven isolates had appressed colony texture, while 3 isolates had velvety and 5 isolates had fluffy. Eight isolates produced aerial mycelium and the other did not. Eight isolates were categorized as fast growing, 8 isolates were categorized as medium growing and 3 isolates were categorized as slow growing. The mean sclerotial sizes of the isolates varied between 65.6 and 127.4 µm. The sclerotia of the 11 isolates were round, 3 isolates were ovoid and 5 isolates were irregular shaped. The size of the PCR products varied from 435 to 518bp and the isolates showed between 99.61 and 100% identity with the reference. The isolates were highly variable in their morphological, molecular and virulence characteristics.

Determination of the Antagonistic Effects of Some Rhizospheric Bacteria against Macrophomina phaseolina under In Vitro Conditions

Macrophomina phaseolina (Tassi) Goid. is a fungal pathogen causes charcoal rot disease (Sin: Rhizoctonia bataticola) and is responsible for significant yield losses in many plants. In our study, we aimed to evaluate the antagonistic ability of 39 different bacteria, isolated from the fields of sugar beet in 2019, against the pathogen Macrophomina phaseolina isolated from sugar beet, beans and chickpeas. Approximately 31% of the bacteria showed antibiosis effect against the pathogen. It was determined that the effectiveness level of Lelliottia amnigena, Bacillus atrophaeus, B.pumilus and B. cereus (7 isolates) was moderate to high against Macrophomina phaseolina. Bacillus atrophaeus (PTo15-1a) showed the highest efficacy of 80%, 72.94% and 82.35% against Macrophomina phaseolina of chickpea, bean and sugar beet respectively. Lelliottia amnigena (Pto 14-1b) was moderately effective (57.78%) against the chickpea isolate of the pathogen. It was observed that of the seven Bacillus cereus isolates used in the experiment, three isolates (Pto14-1a, Pto12-1b, Pto17-1b) were highly effective against the chickpea pathogen, two (Pto12-1b, Pto14-2b) against bean pathogen, and one (Pto15-1b) against sugar beet isolate. Results have shown varied level of antagonism by different test bacterial against different Macrophomina phaseolina isolates, while the highest level of antibiosis shown by Bacillus atrophaeus against all pathogenic isolates indicated that it can be a potential future bioagent in managing the disease.

The full-length genome characterization and diversity of faba bean necrotic stunt virus in Iran

Faba bean necrotic stunt virus (FBNSV) (genus Nanovirus; family Nanoviridae) is a multipartite virus comprising eight circular single-stranded DNA components, each about 1 kb in length. Also, two alphasatellites have often been reported to be associated with FBNSV infection. The only full-length genome sequence of an Iranian FBNSV isolate, FBNSV-[IR;Ka:FB], so far reported is one infecting faba bean (Vicia faba L.). To obtain more information about Iranian isolates of FBNSV (FBNSV-IRs) and their diversity, the full-length genome of four FBNSV isolates and associated alphasatellites obtained from faba bean (two isolates), common bean (Phaseolus vulgaris L., one isolate) and chickpea (Cicer arietinum L., one isolate) were characterized. All genomic components except U4 had the expected size in accordance with the FBNSV genomic sequences published in the GenBank. The component U4 had an additional stretch of 16 nucleotides (nts 765–780) in its non-coding region (NCR). Alphasatellites of the aforementioned isolates showed 98–100% sequence identity. Intra-component sequence analyses of FBNSV-IRs indicated a range of 98 to 100% nt identity between each single FBNSV-IRs component or between their associated alphasatellites. DNA-M and DNA-U2 exhibited the highest value of diversity, sharing 98–99% identity. Phylogenetic analyses indicated that FBNSV-IRs were clustered with a Moroccan isolate and some Azerbaijani isolates of FBNSV forming a clade distinct from Ethiopian isolates. Inter-component sequence analyses of NCRs revealed the DNA-S and DNA-U4 subgrouped together in all FBNSV-IRs isolates except the chickpea isolate in which NCRs of DNA-S and DNA–U4 formed separate subgroups.

Viromes of Ten Alfalfa Plants in Australia Reveal Diverse Known Viruses and a Novel RNA Virus.

Alfalfa plants in the field can display a range of virus-like symptoms, especially when grown over many years for seed production. Most known alfalfa viruses have RNA genomes, some of which can be detected using diagnostic assays, but many viruses of alfalfa are not well characterized. This study aims to identify the RNA and DNA virus complexes associated with alfalfa plants in Australia. To maximize the detection of RNA viruses, we purified double-stranded RNA (dsRNA) for high throughput sequencing and characterized the viromes of ten alfalfa samples that showed diverse virus-like symptoms. Using Illumina sequencing of tagged cDNA libraries from immune-captured dsRNA, we identified sequences of the single-stranded RNA viruses, alfalfa mosaic virus (AMV), bean leafroll virus, a new emaravirus tentatively named alfalfa ringspot-associated virus, and persistent dsRNA viruses belonging to the families Amalgaviridae and Partitiviridae. Furthermore, rolling circle amplification and restriction enzyme digestion revealed the complete genome of chickpea chlorosis Australia virus, a mastrevirus (family Geminiviridae) previously reported only from chickpea and French bean that was 97% identical to the chickpea isolate. The sequence data also enabled the assembly of the first complete genome (RNAs 1–3) of an Australian AMV isolate from alfalfa.

Impact of hydrolysis on functional properties, antioxidant, ACE-I inhibitory and antiproliferative activity of Cicer arietinum and Cicer reticulatum hydrolysates

Chickpea is an important food legume of the world offering valuable nutrients. Employing the degree of hydrolysis (DH) by the enzyme alcalase and flavourzyme, the functional properties of two chickpea species i.e. Cicer arietinum and Cicer reticulatum was investigated. This includes antioxidant, physicochemical properties and angiotensin-converting enzyme (ACE-I) inhibitory activities. Hydrolysis reaction was performed in different time intervals including 0, 20, 40, 60, 80, and 100 min by adding 2% (w/v) alcalase (pH 8) and flavourzyme (pH 7) at 50 °C temperature and pH was kept constant during hydrolysis time. DH improved through solubility of chickpea isolate in the pH range of 5–10. Alcalase produced a maximum degree of hydrolysis at 60 min for both the Cicer species. Flavourzyme demonstrated the highest degree hydrolysis for C. arietinum at 80 min and C. reticulatum at 100 min. The Cicer protein hydrolysates showed the highest antioxidant, ACE-I inhibitory and antiproliferative activity. As a prerequisite to food industry, present study revealed that chickpea protein hydrolysate (CPH) can be appraised as a source of functional food.

English

Author(s): Zafar, M; Ahmed, N; Mustafa, G; Zahir, ZA; Simms, EL | Abstract: There are various mechanisms through which plant’s growth and yield can be promoted by plant growth promoting rhizobacteria (PGPR) that reside inside the soil. The use of pesticides, chemical fertilizers and other supplements could be replaced by PGPR. There are various substances produced by the rhizosphere microorganisms that are important for growth promotion as they affect morphology of plants indirectly. Their nodulation activity, biofilm formation, root colonization were observed and characterized for solubilization of phosphate, production of indole acetic acid and fixation of nitrogen. It was found that ten out of seventeen isolates from chickpea had positive nitrogen fixing activity but at varying levels and the range of fixed nitrogen was 136.666 to 303.666 nmoles C2H2 reduced/h/mg protein. Bacterial isolate CM1 showed maximum nitrogen fixation ability. Thirteen isolates produced IAA in the range of 3.833-18.100 µg/mL. Bacterial isolate, CM1 produced maximum indole acetic acid. Among chickpea bacterial isolates, sixteen were positive in phosphate solubilizing ability while only one was negative. Range of phosphate solubilization among chickpea isolates was 4.533-12.333µg/mL. Maximum value for phosphate solubilization was observed in CM4. Potential root nodulating bacteria were identified using 16S rRNA sequence analysis. CM1, CM2, CM3, CM5, CM6, CM11, CM15, CM24 and CM25 were identified as Mesorhizobium ciceri while CM4 as Mesorhizobium mediterraneum.

Red clover vein mosaic virus-A Novel Virus to New Zealand that is Widespread in Legumes.

Red clover vein mosaic virus (RCVMV) is an important virus of leguminous crops that can cause devastating losses. During a routine survey of legumes conducted on the South Island of New Zealand, RCVMV was found in mixed infections in clover plants with Alfalfa mosaic virus and White clover mosaic virus. The full-length sequence of the New Zealand isolate RCVMV-NZ from clover shared 96% nucleotide sequence identity with a chickpea isolate previously described from Washington (United States). Targeted surveys of pea, faba bean, and pasture crops showed that RCVMV-NZ is widespread on the South Island in New Zealand. This isolate is causing mild if any symptoms on experimental hosts and naturally infected plants.

English

Wet root rot caused by Rhizoctonia solani is one of the important diseases in chickpea worldwide. In the present study, 10 random amplified polymorphic DNA (RAPD) primers were used to assess the molecular diversity of 50 chickpea isolates of R. solani. There was a great diversity among the isolates studied and was in the range of 52 to 93%. The isolates were highly variable in aggressiveness and caused up to 100% wet root rot incidence in chickpea. Accurate detection and identification of plant pathogens are fundamental to plant pathogen diagnostics and management. Therefore, a polymerase chain reaction (PCR) assay was developed for accurate and sensitive detection of R. solani from mycelial DNA and infected chickpea plants. RAPD primer OPA 11 consistently amplified &asymp;1700 base pairs (bp) product in PCR only from the DNA of R. solani isolated from chickpea. The common DNA fragment was sequenced and used to design a pair of oligonucleotide primers amplifying 285 bp sequence characterized amplified region (SCAR). The specificity of the SCAR primers was evaluated. The detection sensitivity of R. solani was 0.5 ng for the genomic DNA and 5 ng for the DNA extracted from infected chickpea root samples. Also, SCAR primer was validated with Q-PCR to detect and quantify R. solani upto 1 pg from infected chickpea root samples. These new SCAR marker are useful for early detection and quantification of wet root rot pathogen in chickpea. Key words: &nbsp;Anastomosis grouping, chickpea, wet root rot, quantitative-PCR (Q-PCR), random amplified polymorphic DNA (RAPD), sequences characterized amplified region (SCAR).

English

Rhizoctonia solani is considered as one of the most destructive soil-and-seed borne plant pathogens infecting various agricultural crops including chickpea. The 50 chickpea isolates of R. solani representing 10 different states of India were variable in hyphal anastomosis reactions and they were grouped into seven AGs as AG1, AG2-2, AG2-2LP, AG2-3, AG3, AG4 and AG5. Genetic diversity of the pathogen was determined by using molecular markers namely, simple sequence repeats (SSR) and rDNA internal transcribed spacer (ITS). A neighbor-joining tree constructed based on the profiles generated by SSR markers grouped the isolates into eight categories. This revealed 90% of genetic similarity among the isolates and partial correlation with reference to their geographical origin and AGs. The isolates were amplified with a set of primers ITS 1 and ITS 4 and they produced a specific band &asymp;650 bp. Low level of (7%) variability was observed in the nucleotide sequences of the ITS regions of these isolates. The phylogenic tree generated from bootstrap neighboring joint analysis grouped the Indian populations of R. solani into two categories. &nbsp; Key words: Chickpea, wet root rot, genetic diversity, simple sequence repeats (SSR), internal transcribed spacer (ITS).

Development of the teleomorph of Ascochyta rabiei on culture media

Ascochyta blight caused by Didymella rabiei (anamorph: Ascochyta rabiei) is an important foliar disease of chickpea in many countries. The fungus is heterothallic and requires the pairing of two compatible mating types for the teleomorph to develop. In nature, the teleomorph only develops on chickpea debris that overwinters on the soil surface in the presence of both mating types. When natural and synthetic agar media were seeded with conidial suspensions of compatible isolates of D. rabiei from Spain and the United States and incubated under favourable conditions for teleomorph development, the teleomorph only developed on 2 % water agar amended with powdered chickpea stems or hot water extracts of chickpea stems, but not on 14 other natural or synthetic media. Ascospore isolates of D. rabiei from pseudothecia that developed on agar media were indistinguishable in cultural and morphological characteristics from isolates of the fungus from chickpea. Production of pseudothecia and ascospores on the best culture medium was always lower than on stem pieces of chickpea straw used as a control treatment. Ascospores discharged from pseudothecia that developed on powdered chickpea stem media onto chickpea seedlings were pathogenic, inducing symptoms identical to those caused by ascospores from chickpea stem pieces or conidia from a chickpea isolate of the fungus. This is the first report of the teleomorph of D. rabiei developing on culture media.

Ascospore dimorphism-associated mating types of Sclerotinia trifoliorum equally capable of inducing mycelial infection on chickpea plants

In many heterothallic fungal pathogens, mating types are found to be associated with variation in virulence and some other ecological traits. Sclerotinia trifoliorum is unique from other Sclerotinia species in that it is heterothallic with two mating types. The mating type gene has pleotropic effect on ascospore size; large ascospore isolates are phenotypically homothallic (L-type), and small ascospore isolates are heterothallic (S-type). The possible association of variation in virulence with the two mating types and hence the ascospore size in S. trifoliorum is investigated using isolates collected from naturally infected chickpea plants and isolates generated from controlled crosses. Chi-square tests showed that 57 field isolates collected from crown lesions (infection initiated by mycelium) had a 1:1 distribution of L-type (29 isolates) and S-type (28 isolates), whereas 14 isolates from stem lesions (infection initiated by ascospores) had a distribution of 10 L-type isolates and 4 S-type isolates not significantly different from 1:1. Greenhouse tests using mycelial plugs as inoculum of field and laboratory-derived isolates did not show significant difference between the two mating types in causing stem rot of chickpea. The sample size of ascospore-initiated infection was small, and the controlled pathogenicity assays in the greenhouse only tested mycelial infection. Thus, whether the two types of ascospores have equal capability of infecting chickpea remains to be further investigated. Strong evidence of both field and greenhouse data showed that mycelia of both mating types of S. trifoliorum were equally capable of infecting chickpea.

Chickpea protein isolates obtained by wet extraction as emulsifying agents

Wet extraction of protein from defatted chickpea (variety Thiva (T), Greece) flour, at alkaline or slightly acidic pH, followed by isoelectric precipitation (pI) or ultrafiltration (UF) to recover the protein, was employed to obtain a number of chickpea protein isolates, enriched either in protein constituents belonging to the globulin (TpI, TUF, TUFG) or to the albumin fraction (TUFA). The interfacial activity and film-forming ability of the isolate protein constituents as well as their emulsifying properties were evaluated. The method applied for chickpea protein isolate preparation influenced to an appreciable extent their composition, adsorption behaviour to oil-water interfaces and emulsion formation and stabilization characteristics, especially with respect to oil droplet flocculation and coalescence. The isolates also differed in their ability to stabilize emulsions subjected to thermal processing or following storage under freezing conditions. The results are discussed in terms of compositional and, possibly, structural differences existing between the protein constituents of the chickpea isolates that may influence their functional behaviour in emulsion systems. The method applied for isolate preparation influenced to an appreciable extent the ability of proteins to adsorb to the oil-water interface and stabilize emulsions during long-time ageing or following heat treatment or freezing.

Determination of genetic diversity among Indian isolates of Rhizoctonia bataticola causing dry root rot of chickpea

Genetic diversity among 27 isolates (23 from chickpea and 4 from other host crops) of Rhizoctonia bataticola representing 11 different states of India was determined by random amplified polymorphic DNA (RAPD), internal transcribed spacer restriction fragment length polymorphism (ITSRFLP) and ITS sequencing. The isolates showed variability in virulence test. Unweighted paired group method with arithmetic average cluster analysis was used to group the isolates into distinct clusters. The clusters generated by RAPD grouped all the isolates into six categories at 40% genetic similarity. High level of diversity was observed among the isolates of different as well as same state. Some of the RAPD (OPN 4, OPN 12, and OPN 20) markers clearly distinguished majority of the isolates into the area specific groups. The ITS I, 5.8 rDNA and ITS II regions of 11 isolates representing different RAPD groups were amplified with primers ITS 1 and ITS 4 and digested with seven restriction enzymes. The restriction enzymes DraI, MboI, RsaI, and AluI were found to be suitable for differentiating the isolates into five categories by showing isolate specific ITS-RFLP patterns. The isolates were variable in their nucleotide sequences of the ITS regions. This is the first study on genetic diversity among chickpea isolates of R. bataticola.

Inefficient nodulation of chickpea (Cicer arietinum L.) in the arid and Saharan climates in Tunisia bySinorhizobium meliloti biovarmedicaginis

The aim of this work is to study the genetic diversity and the symbiotic effectiveness of the natural populations of rhizobia nodulating chickpea (Cicer arietinum L.) in six locations of South Tunisia, where chickpea had never been cultivated. Nodules were observed only in the two soil samples from Gafsa (0.8 nodules per plant) and Tataouine (2 nodules per plant). PCR-RFLP typing of 165 rRNA genes of 42 isolates indicated that all analysed strains showed the same ribotype as the reference strainSinorhizobium meliloti RCR2011. These isolates induced ineffective nodules on chickpea andMedicago sativa; however nodules onMedicago laciniata were effective. Analysis of the symbiotic diversity by PCR-RFLP, of thenifDK spacer suggested that all chickpea isolates from the South belong to the biovarmedicaginis ofS. meliloti. The present paper is, to our knowledge, the first report showing that chickpea is selectively nodulated under soil conditions by a specific biovar ofS. meliloti showing specificity toM. laciniata. The specificity of this interaction as well the impact of this inefficient nodulation on chickpea cultivation needs to be investigated further.

Strains of Mesorhizobium amorphae and Mesorhizobium tianshanense, carrying symbiotic genes of common chickpea endosymbiotic species, constitute a novel biovar (ciceri) capable of nodulating Cicer arietinum

To identify several strains of Mesorhizobium amorphae and Mesorhizobium tianshanense nodulating Cicer arietinum in Spain and Portugal, and to study the symbiotic genes carried by these strains. The sequences of 16S-23S intergenic spacer (ITS), 16S rRNA gene and symbiotic genes nodC and nifH were analysed. According to their 16S rRNA gene and ITS sequences, the strains from this study were identified as M. amorphae and M. tianshanense. The type strains of these species were isolated in China from Glycyrrhiza pallidiflora and Amorpha fruticosa nodules, respectively, and are not capable of nodulating chickpea. These strains carry symbiotic genes, phylogenetically divergent from those of the chickpea isolates, whose nodC and nifH genes showed more than 99% similarity with respect to those from Mesorhizobium ciceri and Mesorhizobium mediterraneum, the two common chickpea nodulating species in Spain and Portugal. The results from this study showed that different symbiotic genes have been acquired by strains from the same species during their coevolution with different legumes in distinct geographical locations. A new infrasubspecific division named biovar ciceri is proposed within M. amorphae and M. tianshanense to include the strains able to effectively nodulate Cicer arietinum.

An Improved Antiserum for Sensitive Serologic Detection of Chickpea chlorotic dwarf virus

AbstractA Syrian chickpea isolate of Chickpea chlorotic dwarf virus (CpCDV; genus Mastrevirus, family Geminiviridae) was purified and yielded 0.6–0.8 mg of purified virus per kg of infected chickpea tissue. The purified preparations were injected into a rabbit and an antiserum of good quality was obtained and used to evaluate different serological tests for the detection of CpCDV in infected chickpea leaf tissue and extracts. CpCDV was detected in sap dilutions of 1/640 by double‐antibody sandwich enzyme‐linked immunosorbent assay (DAS‐ELISA) and dot‐blot ELISA, and in sap dilutions of 1/1280 by direct antigen‐coating (DAC)‐ELISA using CpCDV immunoglobulin G (IgG) at 0.5 μg/ml. The antiserum was also able to detect the capsid protein of CpCDV by Western blot using raw antiserum at a dilution of 1/2000. The CpCDV raw antiserum (third bleeding) produced had a titre of 1/320 000 when determined by tissue‐blot immunoassay (TBIA); whereas, coating ELISA plates with CpCDV IgG at a concentration of 0.004 μg/ml was enough to detect the virus by DAS‐ELISA in a sap dilution of 1/20 using an enzyme conjugate at a dilution of 1/2000.