Immunological detection of bean common mosaic virus in French bean (<Emphasis Type="Italic">Phaseolus vulgaris</Emphasis> L.) leaves

Bean common mosaic potyvirus (BCMV) is an important seed borne pathogen of French bean. Differential inoculation with bean common mosaic virus at cotylodonary trifoliate leaf stage and pre-flowering stage of crop growth revealed that cotyledonary leaf infection favored maximum disease expression. Under immunosorbent electron microscopy (ISEM) the virus particles of filamentous structure having a diameter of 750 nm (l) and 15 nm (w) were observed. These particles gave positive precipitin tests with polyclonal antiserum of Potato virus Y.     

SSR analysis of genetic diversity and structure of the germplasm of faba bean (Vicia faba L.)

Assessing the diversity and genetic structure of faba bean (Vicia faba L.) germplasm is essential to improve the quality and yield of this economically important crop. In this study, simple sequence repeats (SSRs) were utilized to evaluate the diversity and structure of 35 faba bean genotypes originating from three different geographical regions (Northern Africa, Eastern Africa, and Near East). All 15 SSR loci generated a total of 100 alleles. The allele number per locus varied from 4 to 11, with a mean of 6.67. The expected heterozygosity (H_e) of SSR loci ranged between 0.51 and 0.81, with a mean of 0.63. The PIC value also varied from 0.44 to 0.78, with an average of 0.58. The expected heterozygosity of 22 faba bean genotypes was higher than the observed one. Interestingly, AMOVA analysis showed that much of variability resided within accessions (79.2%). A highly significant difference among regions was also evidenced, and represented 5.3% of the total variation. Moreover, cluster analysis divided the 35 faba bean genotypes into two main clusters. The first main cluster comprised all faba bean genotypes originating from the Near East region, whereas the second main cluster comprised all the genotypes originating from the Northern and Eastern Africa regions, indicating that the Northern and Eastern African faba bean genotypes were more closely related to each other than to the Near East genotypes. Structure analysis also revealed that the 35 faba bean genotypes might be assigned to two populations, in complete accordance with cluster analysis data. In conclusion, this study showed high levels of diversity in the analysed genotypes of faba bean, and could be utilized in future breeding programmes to develop new cultivars of high yield.     

Properties of a host range and coat protein variant of broad bean true mosaic comovirus from Vicia faba

Unlike other described isolates of broad bean true mosaic comovirus (BBTMV), a variant, code name SB, infected some non-leguminous plant species and, in N. benthamiana, induced systemic mottling and puckering of the leaves. However, like other described BBTMV isolates, purified SB particle preparations contained isometric particles c. 28 nm in diameter that sedimented as two nucleoprotein components with S_{20, w} values of 90S and 109S; some preparations occasionally contained a component of c. 50S. Virus particles contained two ssRNA species which, when denatured in glyoxal, had estimated M_T values of 2.1 × 10⁶ and 1.3 × 10⁶ and co-electrophoresed with cowpea mosaic virus RNA-1 and RNA-2 respectively. Isolate SB was serologically indistinguishable from British and German isolates of BBTMV. However, SB virus particles contained a major polypeptide (L) of M_r between c. 31 000 and up to three minor ones (S) or M_r between c. 20 000 and 24 000. This contrasts with protein preparations from other BBTMV isolates that typically contain only two polypeptides of M_r c. 37 000 (L) and 21 000 (S). Following isopycnic centrifugation in CsCl, SB particles purified from pea separated into two major components with densities of 1.39 and 1.44 g cm^-3 and a minor component of estimated density 1.43 g cm^-3. In Cs₂SO₄, virus preparations separated into three major components with densities of 1.30, 1.32 and 1.36 g cm^-3 and a minor one of density 1.27 g cm^-3. In CsCl isopycnic gradients, SB particles purified from TV. benthamiana separated into two components with densities of 1.38 and 1.43 g cm^-3. During immuno-electrophoresis in agarose gels, freshly prepared virus and preparations stored for up to 4 days at 4°C contained a single component that migrated rapidly to the anode, whereas similar preparations of an English isolate of BBTMV migrated as a single component that moved only slowly toward the anode but which, within 48 h, contained an additional component with a migration rate similar to that of isolate SB. Isolate SB is therefore a host range variant of BBTMV which, in comparison with previously described isolates of BBTMV, has an increased negative charge of its particles prior to any appreciable degradation of its S protein, and S protein that is degraded less rapidly. These features probably account for the anomalies observed in isopycnic centrifugation.     

Nutritional and antinutritional attributes of faba bean (Vicia faba L.) germplasms growing in Bihar,India

Eleven germplasms of faba bean seeds from four agroclimatic regions of Bihar, India, have been investigated to estimate their nutritional (soluble protein, free amino acids, starch, reducing and non reducing sugar, total soluble sugar) and antinutritional (total extractable phenol and condensed tannin/proanthocyanidin) parameters. These parameters were found in varying concentration in all genotypes studied. The highest concentration of total extractable phenol and proanthocyanidin (condensed tannin) (2.56 and 1.59 % leucocyanidin equivalents respectively on dry matter basis) were found in Samastipur while the lowest from Patna (0.95 and 0.426 % leucocyanidin equivalent on dry matter basis). The different nutritional parameters were also found to be in variable concentration among different germplasms viz. total soluble protein ≈ 20–32 %, free amino acids ≈ 188–348 mg/100 g, starch ≈ 27–33 %, reducing sugars ≈ 85–188 mg/100 g, non reducing sugars ≈ 0.7–1.7 % and total soluble sugars ≈ 0.8–1.9 %.     

Phylogenetic multilocus sequence analysis of native rhizobia nodulating faba bean (Vicia faba L.) in Egypt

The taxonomic diversity of forty-two Rhizobium strains, isolated from nodules of faba bean grown in Egypt, was studied using 16S rRNA sequencing, multilocus sequence analyses (MLSA) of three chromosomal housekeeping loci and one nodulation gene (nodA). Based on the 16S rRNA gene sequences, most of the strains were related to Rhizobium leguminosarum, Rhizobium etli, and Rhizobium radiobacter (syn. Agrobacterium tumefaciens). A maximum likelihood (ML) tree built from the concatenated sequences of housekeeping proteins encoded by glnA, gyrB and recA, revealed the existence of three distinct genospecies (I, II and III) affiliated to the defined species within the genus Rhizobium/Agrobacterium. Seventeen strains in genospecies I could be classified as R. leguminosarum sv. viciae. Whereas, a single strain of genospecies II was linked to R. etli. Interestingly, twenty-four strains of genospecies III were identified as A. tumefaciens. Strains of R. etli and A. tumefaciens have been shown to harbor the nodA gene and formed effective symbioses with faba bean plants in Leonard jar assemblies. In the nodA tree, strains belonging to the putative genospecies were closely related to each other and were clustered tightly to R. leguminosarum sv. viciae, supporting the hypothesis that symbiotic and core genome of the species have different evolutionary histories and indicative of horizontal gene transfer among these rhizobia.     

BCMV-ukr: isolate of bean common mosaic virus revealed in Ukraine

SUMMARY

Bean common mosaic virus (BCMV) is distributed worldwide and causes a serious disease in bean reducing growth and crops yield. The aim of this study was to investigate the occurrence of BCMV and Bean common mosaic necrotic virus (BCMNV) in Ukraine, to characterise host range and reactions of indicator plants to mechanical inoculation with the isolate and to differentiate it by using Drijfhout’s differentials. Leaf samples were positive for BCMV infection in RT-PCR assay employing specific primers with amplification of a 340-bp product. Based on a biological test on bean differentials, the isolate was assigned to pathogroup VII despite the fact that strain differed markedly from the standard strains in symptoms producing on differential groups IV and V. Partial sequence data of the coat protein region show 100% identity with BCMV 125 sequences tested. To our knowledge, this is the first attempt to characterise the BCMV circulating in Ukraine.     

Management of jute yellow mosaic virus disease through cultural practices

A field experiment was conducted to investigate the effect of some management practices to minimise jute yellow mosaic virus disease. The management practices were employed at natural condition and placed randomly with four replications. The treatments were spraying malathion 57 EC, rouging and field sanitation, top dressing of nitrogenous fertiliser, mulching with straw and untreated control. The highest percentage of mosaic incidence was recorded in control and the lowest incidence was recorded in top dressing of nitrogenous fertiliser. Among the treatments, top dressing of nitrogenous fertiliser showed the best performance in terms of increasing yield (3.05?t/ha). The second highest was obtained in rouging and field sanitation which was statistically similar to spraying malathion 57 EC. The best gross margin ($379.02/ha) and increase of gross margin (63.00%) compared to control were achieved in top dressing of nitrogenous fertiliser with the highest benefit-cost ratio (4.84). However, the treatments were found significantly profitable compared to the control indicating the usefulness of the cultural practices in integrated disease management programme for healthy and profitable jute cultivation.     

Cytopathology of soybean tissue culture cells infected with southern bean mosaic virus

Summary Virus distribution patterns and ultrastructural changes in soybean callus cells after infection with the type, or bean strain, of southern bean mosaic virus (SBMV) were observed. Calli grown in liquid Linsmaier and Skoog (LS) medium were inoculated with SBMV and incubated in fresh LS medium. Calli were sampled at 5, 10, 15, and 20 d after inoculation and examined by transmission electron microscopy. Five days after inoculation, viruslike particles (VLP) measuring 22 to 27 nm in d were observed in the cytoplasm. The particles formed loose aggregates with some tendency to associate in regular patterns. By the 10th d after infection, particles were observed in the vacuoles in similar loose arrangements. Viruslike particles were readily identified in vacuoles because of the absence of ribosomes. Crystalline aggregations of VLP were found from Day 10 to Day 20 in the cytoplasm only. Five days after inoculation particles similar to the VLP observed in the cytoplasm also were present in nuclei. Other cytopathic effects were noted, particularly several types of inclusion bodies. These observations differ considerably from reports of the type strain in intact bean plant tissues in the frequent occurrence of VLP in vacuoles and virus crystals in cytoplasm of soybean callus infected with the same strain of virus. Michigan Agricultural Experiment Station Journal Article Number 10020. We thank Dr. Karen Baker for useful suggestions.     

Production of Bean yellow mosaic virus resistant subterranean clover (Trifolium subterraneum) plants by transformation with the virus coat protein gene

Transgenic lines of subterranean clover were constructed that contained three different Bean yellow mosaic virus (BYMV) coat protein (CP) gene constructs; full-length CP, the core region of the CP, and full-length CP plus the 3′ untranslated region of the viral genome. Transgenic plants containing the full-length and core CP gene constructs showed high and moderate levels of BYMV resistance. Resistance was measured as a lack or amelioration of viral disease symptoms, which was correlated with a reduction in virus levels and yield loss. A range of different resistance phenotypes was observed. They included reduced infection rates, delay and reduction in local lesion development, and delay and reduction in severity of systemic symptom development. Resistance levels were not correlated with transgene mRNA levels and no transgene-encoded protein was detected in any of the transgenic lines. This is the first example of genetically engineered virus resistance in a clover.     

Analysis of nucleotide sequence of wheat yellow mosaic virus genomic RNAs

Wheat yellow mosaic virus (WYMV) isolate HC was used for viral cDNA synthesis and sequencing. The results show that the viral RNA1 is 7629 nueleotides encoding a polyprotein with 2407 amino acids, from which seven putative proteins may be produced by an autolytie cleavage processing besides the viral coat protein. The RNA2 is 3639 nueleotides and codes for a polypretein of 903 amino acids, which may contain two putative non-structural proteins. Although WYMV shares a similarity in genetic organization to wheat spindle streak mosaic virus (WSSMV), the identities in their nucleotide sequences or deduced amino acid sequences are as low as 70% and 75 % respectively. Based on this result, it is confirmed that WYMV and WSSMV are different species within Bymovirus.     

Molecular Analysis of Zucchini yellow mosaic virus Isolates from Hangzhou, China

Isolates of Zucchini yellow mosaic virus were obtained from different cucurbit crops in Hangzhou city, China. The complete nucleotide sequences of four isolates and the 3′‐terminal sequences, including the coat protein coding region, of four others were determined and then compared with other available sequences. Phylogenetic analysis of the coat protein nucleotide sequences showed that these isolates fell into three significant groups, one of which (designated group III) consisted exclusively of Chinese isolates and is reported for the first time. Comparisons over the completely sequenced genomes showed that, typically for potyviruses, the 5′‐end of the genome was usually the most variable but that the group III isolate differed from the others most significantly in the N‐terminal part of the coat protein. Partially sequenced group III isolates also varied from other isolates in this region. Group III isolates appear to differ biologically from the other isolates because they do not cause symptoms in watermelon fruit but induce more severe symptoms on the watermelon leaves.     

Propyl gallate,a free radical scavenger,counteracts the benefits of exogenously applied salicylic acid and aggravates the deleterious effects of the southern bean mosaic virus in Rhizobium-nodulated Phaseolus vulgaris plants

Infection of Rhizobium-nodulated Phaseolus vulgaris by the southern bean mosaic virus (SBMV) markedly decreased the growth and nodulation of plants. Exogenous applications of salicylic acid (SA) at concentrations ≥10 μM further decreased growth and nodulation of virus-infected (V) plants. Only SA concentration of 5 μM in the solution improved the growth, nodulation, chlorophyll concentration and the catabolism of ureide in leaves of V plants. The spray of leaves with 2 mM propyl gallate (+Pg) decreased growth, nodulation as well as the chlorophyll and leaf ureide concentrations in V plants, regardless of the concentration of SA at which plants were grown. Ultrastructural damages in leaf cells caused by SBMV were also enhanced in V+Pg plants. The massive proliferation of virus particles and the presence of virus crystalline arrays within symbiosomes of nodules in V+Pg plants were attributed to proliferation of branched plasmodesmata in leaf vascular-parenchyma cell walls that facilitated virus movement. Virus particles were never observed in leaf and nodule tissues of V plants not sprayed with Pg. Exogenous applications of SA hindered while Pg increased the symbiotic performance of H plants, pointing out the complexity to be addressed in breeding for virus resistance in Rhizobium-nodulated beans.     

Morphological and physiological characterization of different genotypes of faba bean under heat stress

Heat stress (HS) is the major constraint to crop productivity worldwide. The objective of the present experiment was to select the tolerant and sensitive genotype(s) on the basis of morpho-physiological and biochemical characteristics of ten Vicia faba genotypes. These genotypes were as follows: Zafar 1, Zafar 2, Shebam 1, Makamora, Espan, Giza Blanka, Giza 3, C4, C5 and G853. The experimental work was undertaken to study the effects of different levels of temperature (control, mild, and modest) on plant height (PH) plant⁻¹, fresh weight (FW) and dry weight (DW) plant⁻¹, area leaf⁻¹, content of leaf relative water (RWC), proline content (Pro) and total chlorophyll (Total Chl), electrolyte leakage (EL), malondialdehyde level (MDA), hydrogen peroxide (H₂O₂), and activities of catalase (CAT), peroxidase (POD) and superoxide dismutase (SOD) enzymes. HS significantly affected growth performance of all genotypes. However, the magnitude of reduction in genotypes ‘C5’ was relatively low, possibly due to its better antioxidant activities (CAT, POD and SOD), and accumulation of Pro and Total Chl, and leaf RWC. In the study, ‘C5’ was noted to be the most HS tolerant and ‘Espan’ most HS sensitive genotypes. It was concluded that the heat-tolerant genotypes may have better osmotic adjustment and protection from free radicals by increasing the accumulation of Pro content with increased activities of antioxidant enzyme.     

Enhancement of resistance by poultry manure and plant hormones (salicylic acid & citric acid) against tobacco mosaic virus

Virus is the most menacing factor for plant, which causes enormous economic losses in agriculture worldwide. Tobacco mosaic virus is most hazardous virus among the plants that can spread through biological and non-biological sources. TMV is ancient virus that causes huge economic losses to pepper cucumber ornamental crops and tobacco. It can be controlled by reducing the population of vector through pesticide application. However, the rapid usage of synthetic chemicals causes environmental pollution and destroys our ecosystem. Consequently, different approaches just like natural derivatives should be adopted for the environmental friendly management for TMV. This in vitro study demonstrated the potential role of natural metabolites such as poultry manure and plant extracts such as salicylic acid and citric acid for the control of TMV. Two different concentrations of poultry manure 60G and 30G were used. Poultry manure was mixed with the soil at the time of sowing. Disease severity was minimum at maximum concentration as compared to the control. Meanwhile, two different concentrations of salicylic acid and citric acid 60% and 90% were applied by foliar sprayer after three-leaf stages. Disease severity was observed after 5, 10, 15, 20, 25, and 30 days after disease inoculation. Here also maximum concentration showed the minimum disease severity and higher concentration of both animal and plants extracts were used for following experiment. Quantitative real-time PCR (RT-qPCR) results demonstrated that different plant defense-related genes such as PR1a, PAL, PR5, NPR1, PRIb, and PDF1.2 were up-regulated. Furthermore, applications of each treatment-induced systemic resistance against a wide range of pathogen including TMV and fungal pathogen Botrytis cinerea.     

Completion of the Nucleotide Sequence of a Brazilian Isolate of Southern bean mosaic virus

The sequence of the central part (ORF2) of a Brazilian isolate of Southern bean mosaic virus (SBMV^SP) is described. This ORF is 2888 nt long and together with the previously-sequenced 5' and 3' ends provides the complete nucleotide sequence of this virus isolate. The SBMV^SP RNA encodes four overlapping open reading frames (ORF1, ORF2a, ORF2b, ORF4) and has a genome organization similar to that of the Cocksfoot mottle sobemovirus .     

An inhibitor in faba bean sap suppressing infection of Chenopodium quinoa by bean yellow mosaic virus

An inhibitor in faba bean sap decreased the infectivity of bean yellow mosaic virus in undiluted sap to 0.2% of its potential infectivity calculated from diluted preparations.