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Revolutionize in Protein Patterns of Different Vicia faba L. Cultivars Infected with Broad bean true mosaic virus under Salicylic Acid Treatments



Essam K. Elbeshehy, Ahmed M. Elazzazy and Omar A. Almaghrabi
 
ABSTRACT

A variety of faba bean cultivars infected with BBTMV showed symptoms as severe mosaic, mottling, crinkling, size decline and malformation. Salicylic Acid (SA) application before inoculation specially 100 μM concentrate was reduced the virus symptoms. In Infected plants under all SA treatments were decreased the viral concentration, percentage of infection and severity of diseases compared with decreasing under BBTMV infection without SA treatments. Increase the level of SA play role important for resistance against BBTMV infection in all faba bean cultivars. Possessions of viral disease with SA application were tested after 21 days from inoculation measured up to healthy control on some protein characters include the quantity of protein bands, molecular weight of band and percentage of the protein band in all diverse faba bean cultivars within this investigation. The results showed that all SA treatments increased all properties of protein patterns. The highest increase was observed when seedling treated by 100 μM (SA and BBTMV) compared with uninfected control. The highest increase of protein contents was observed in infected cv. Sakha1 and lower increase was noticed in cv. Tribe White. In the largest parts of SA applications, the all protein characters were mentioned above an increased includes protein amounts and numbers. Total soluble proteins were accumulated when healthy plants were treated by SA. Stimulation of broad bean cultivars to BBTMV infection under SA application were observed in found many modifications in protein patterns. Probably, the polypeptides accumulation is depending on the SA level. The main purpose of this research was to invistegate the effects of BBTMV and SA treatments on symptoms development, virus concentrations, percentage of infections, disease severity, protein patterns in different broad bean leave cultivars, protein bands numbers, molecular weight bands and percentage of amounts in susceptible and tolerance broad bean cultivars.

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Essam K. Elbeshehy, Ahmed M. Elazzazy and Omar A. Almaghrabi, 2014. Revolutionize in Protein Patterns of Different Vicia faba L. Cultivars Infected with Broad bean true mosaic virus under Salicylic Acid Treatments. Plant Pathology Journal, 13: 71-86.

DOI: 10.3923/ppj.2014.71.86

URL: https://scialert.net/abstract/?doi=ppj.2014.71.86
 
Received: December 09, 2013; Accepted: March 07, 2014; Published: April 19, 2014

INTRODUCTION

Faba bean, Vicia faba L. (Fabaceae), is considered as one of the most important food legumes in Egypt. The faba bean is a major staple food crop in Africa. Farmers plant around four million hectares of beans which correspond to 20% of total crop area planted (Bond, 1987). Broad beans can get better soil productiveness and decline the incident of infections, under certain environmental conditions (Mwanamwenge et al., 1998). A number of viruses may cause severe diseases in faba bean crops. There are 15 different viruses in the genus Comovirus of the family Comoviridae, such as BBTMV. Vicia faba L. one of the most important host plant to BBTMV, including other legume plants like peas, (Quantz, 1953). Symptoms of BBTMV infection can be severe in cool includes mild mosaic, malformation and necrosis (Paul and Quantz, 1959). Salicylic Acid (SA) is a natural signaling molecule involved in plant defense response against pathogen infection. Its function in plant disease resistance is well recognized for plants, where it is required for basal resistance against pathogens as well as for the inducible defense mechanism, Systemic Acquired Resistance (SAR) which confers resistance against a broad-spectrum of pathogens. (Chaturvedi and Shah, 2007). Our study appraised the special effects of exogenous treatment SA on the disease severity, BBTMV infection percentage, protein patterns in cultivars, protein bands numerals, molecular weight for each band and percentage of bands in susceptible and tolerance broad bean cultivars.

MATERIAL AND METHODS

The DAS-ELISA technique was applied as described by Clark and Adams (1977) to measure BBTMV concentrations in the infected and (SA+V)-treated leaves using three antibodies BBTMV, BBSMV and BYMV were obtained from International center for Agricultural Research in the Dry Areas (ICARDA), Plant virology, P.O. Box 5466, Aleppo, Syria.

Four cultivars of faba bean, cv. Tribe White, Giza 3, Giza 717 and Sakha l) were planting in natural and favorable conditions suitable for broad bean growth. BBTMV used in these experiments was prepared from fresh severely infected leaves of faba bean cv. Giza 3. After 21 days of growth, plants with similar size were selected and divided into five groups. Each group consists of four replicates (a replicate is one pot containing three healthy plants). The namely of the groups was as follows, group 1: Control (sprayed with water), group 2: Inoculated with virus at the same time with the other groups, group 3: 50 μM (SA and BBTMV), group 4: 100 μM (SA and BBTMV), sprayed by SA and inoculated with virus 3 days later (SA and BBTMV) and group 5: 100 μM SA, sprayed without followed virus inoculation.

Every part of plant leaves were sprayed by SA treated. All leaves were mechanically inoculated by BBTMV inoculation after three days from SA spraying (Radwan et al., 2008). Disease severity and percentage of infection were calculated after 21 days from seedling inoculation and were recorded according to the subsequent balance (0) No external symptoms, (1) Light mottling and crinkling, (2) Mottling and crinkling, (3) Strict mosaic, size diminution and (4) Deformation. Disease Severity (DS) values were calculated using the following formula according to Yang et al. (1996):

The youngest developed leaves from both control and treated plants from four cultivars broad bean were collected after three weeks from inoculation for analysis of changes, 50 mg dry tissues of each cultivar from four broad bean leave cultivars infected and uninfected by BBTMV were treated with different salicylic acid concentrations and compared with control. the leaves cultivars were crocheted and extracted with SDS reducing buffer to determine the total soluble protein contents.

SDS-PAGE procedure was portrayed by Laemmli (1970). Gels were photographed and scanned by Gel Doc Bio-Rad System (Gel-Pro analyzer V3) t determine the percentage of similarity of four broad bean leaves cultivars of each treatment group and electrophoretic profile of total soluble protein extracted from treated and untreated four broad bean leaved cultivars.

ANOVA type one-way were used to calculated the noteworthy diversity in the averages of the experimental treatments. A probability at level of 0.05 or less was measured considerable.

RESULTS AND DISUSSION

The leaf cultivars of broad bean (Tribe White, Giza 3, Giza 717 and Sakha l) were exhibited systemic rigorous symptom under BBTMV inoculation compared with non-inoculated. Novel symptoms appeared including mosaic and crinkling, size diminution and malformation in susceptible cultivars cv. Giza 3 and Tribe White (Fig. 1), other than, mild mosaic, size reduction and light mottling in tolerant cultivars CV. Giza 717 and Sakha l (Fig. 2). On the other hand, SA treatments diminished the exterior of damaging symptoms reasoned by virus progress, principally when the leaves were treated with 100 μM SA under BBTMV infection while 100 μM (SA+V) treated leaves had no celebrate symptoms. On the other hand, mild BBTMV symptoms were observed by 50 μM (SA+V) application (Radwan et al., 2007;Radwan et al., 2008). This type of the systemic resistance might be initiated by SA is spraying in broad bean plants against BYMV Elbadry et al. (2006).

The products of several disobedience genes were triggered appearance and connected with plant tolerance, possibly will playing significant responsibility in the constraint of pathogen development and dispersal. Typical characteristic indicator genes of Systemic Acquired Resistance (SAR) are for instance encoding pathogenesis-related protein PR1 (Van Loon and Van Kammen, 1970; Fraser, 1982; Van Loon et al., 1994; Van Loon and Van Strien, 1999).

Results showing in Fig. 3a-c, revealed that BBTMV infection inoculated plants have higher percentage of infection, severity of disease and concentration of virus while lower in case of SA treated plants and control. This regular lessen of infection percentage, severity of disease and concentration of virus with an increase level of SA play role important for resistance against BBTMV infection in all cultivars of faba bean compared with untreated one. Radwan et al. (2008) also recorded that SA treatments reduced the concentration of BYMV, infection percentage and severity of disease.

The results tabulated in (Table1and 2) and illustrated in (Fig. 4) showed that the susceptible cultivars cv. Giza 3 and Tribe White were recorded a low number of protein bands in all treated groups (6, 4; 6, 6; 5, 6; 7, 10 and 6, 8 PBs), respectively, when inoculated by BBTMV under SA treatments compared with control.

Fig. 1(a-b): Stimulation of susceptible faba bean leaf cultivars cv. Giza 3 and Tribe White to BBTMV infection showing mosaic and crinkling, size diminution and malformation compared with control (a) Susceptible cultivars cv. Giza 3 and (b) Tribe white

Their Molecular Weights (MW) were arranged between 84 to 28 KDa in cv. Giza 3 and arranged between 162 to 29 KDa in cv. Tribe white. While, The results tabulated in (Table 3 and 4) and illustrated in (Fig. 5) showed that the tolerance cultivars cv. Giza 717 and Sakha l were recorded high number of protein bands (7, 11; 8, 12; 9, 12; 10, 14 and 8, 11 PBs) respectively, in all treated groups when inoculated by BBTMV under SA treatments compared with control and (MW) were arranged between 265 to 28 KDa in cv. Giza 717 and arranged between 157 to 33 KDa in cv. Sakha1 and these results are in agreement with (Radwan et al., 2007).

Stimulation of infected broad bean under SA application on percentage of protein band amount (% of Amo.) in difference faba bean leave cultivars were presented in (Table 1-4) and illustrated in (Fig. 4 and 5). In susceptible Giza 3 and Trible white cultivars (% of Amo.) arranged between 1.758-18.257 and 3.245-16.349% in treated group1; 6.032 to 17.825 and 1.996-14.289% in treated group 2; 6.658-15.024 and 1.987-17.289% in treated group 3; 7.008-21.929 and 1.720 to 21.389% in treated group 4 plus 6.203-18.937 and 1.542-19.242% in treated group 5, respectively while, in tolerance Sakha1 and Giza 717 cultivars (% of Amo.) arranged between 2.417-21.133 and 5.853-16.239%; 3.634-23.695 and 5.397-21.341%; 3.663-26.038 and 5.702-23.750%; 3.484-35.996 and 5.990 to 35.451% also 3.581-30.193 and 3.512-32.747%, respectively. Therefore, the percentage of protein amount bands in infected lanes were less than that of the control in cv. Tribe white and Giza 3 but, more than in cv. Sakha 1 and Giza 717.

Fig. 2(a-b): Stimulation of tolerance cultivars cv. Giza 717 and Sakha l to BYMV infection showing mild mosaic, size reduction and light mottling compared with control (a) Tolerance cultivars cv. Giza 717 and (b) cv. Sakha 1

On the other hand, it can be distinguishable that sole bands include a high percentage of the protein band amount were presented in infected cultivars under all treatments by salicylic acid and absented in control healthy such as band number 9 (MW. 52 and RF. 0.699) of cv. Giza 3 absent in treated group 1(control healthy) and was recorded 12.321, 13.538, 15.822 and 13.367% in treated groups (2-5), correspondingly. Alternatively, band number 7 (MW. 55 and RF. 0.659) present in healthy control group 1 was recorded 12.862% and absent in treated group (2-5).

The bands number 16 (MW. 50 and RF. 0.737) and 19 (MW. 42 and RF. 0.803) of cv. Tribe white were absent in treated group1 and were recorded (14.591, 13.137, 17.439 and 13.367%) and (13.755, 17.289, 21.389 and 19. 242%) in treated group (2-5), in that order. Conversely, bands number 17 (MW. 47 and RF. 0.757) and 20 (MW. 41 and RF. 0.817) present in healthy control group 1 were recorded 19.296% and 16.349, respectively and absent in treated group (2-5).

In addition to, the bands number 24 (MW. 115 and RF. 0.366), 39 (MW. 54 and RF. 0.685) and 44 (MW. 33 and RF. 0.953 of cv. Sakha1 were absent in treating group1 and were recorded (17.987, 26.038, 26.049 and 24.095%), (23.695, 24.461, 26.921 and 25.947%) and (16.779, 19.631, 29.321 and 19. 956%) in treated group (2-5), correspondingly. On the other hand, bands number 27 (MW. 102 and RF. 0.413) and 32 (MW. 76 and RF. 0.525) present in healthy control group 1 were recorded 16.071% and 21.133, respectively and absent in treated group (2-5).

Finally, the bands number 10 (MW. 83 and RF. 0.548), 14 (MW. 65 and RF. 0.651) and 15 (MW. 58 and RF. 0. 698 of cv. Giza 717 were absent in treated group1 and were recorded (21.341, 21.680, 28.538 and 27.998%), (13.234, 19.896, 33.047 and 21.063%) and (17.974, 17.721, 30.570 and 26.080%) in treated group (2-5), respectively. Then again, bands number 9 (MW. 85 and RF. 0.536) and 20 (MW. 49 and RF. 0.772) present in healthy control group 1 were recorded 14.265% and 15.065, respectively and absent in treated group (2-5).

Orientation of lofty quantity of proteins was attended to cooperate a significant position in resistance reply in opposition to viruses. Van Loon (1985) confirmed that in more than a few plants groups infected by many viruses the expansion of symptoms is go together with manifestation of novel proteins, whose incidence from cooperation between specific pathogen and host origin.

Table 1: Effect of salicylic acid treatments on protein bands in infected faba bean Giza 3 cultivar after 21 days from inoculation by BBTMV

Table 2: Effect of salicylic acid treatments on protein bands in infected faba bean Tribe white cultivar after 21 days from inoculation by BBTMV

Table 3: Effect of salicylic acid treatments on protein bands in infected faba bean Sakha1 cultivar after 21 days from inoculation by BBTMV

Table 4: Effect of salicylic acid treatments on protein bands in infected faba bean Giza717 cultivar after 21 days from inoculation by BBTMV

Fig. 3(a-c): Virus concentration, percentage of infection and diseases severity between faba bean cultivars, BBTMV and SA treatments calculated of (a) Virus concentration between faba bean cultivars, BBTMV and SA treatments, (b) Diseases severity between faba bean cultivars, BBTMV and SA treatments and (c) Percentage of infection between faba bean cultivars BBTMV and SA treatments




Fig. 4(a-f): Electrophoretic profile of total soluble protein extracted from treated and untreated four susceptible cultivars cv. Giza 3 and Tribe White under five treatment groups. (M) Protein marker



Fig. 5(a-f): Electrophoretic profile of total soluble protein extracted from treated and untreated four tolerance cultivars cv. Giza 717 and Sakha l under five treatment groups. (M) Protein marker

Bean plants infected with BYMV were recorded higher percentage of protein contents compared with healthy plants. As a general rule of SA application were observed an increase in protein amounts (Radwan et al., 2010). PVY infection was recorded higher whole comfortable of PR proteins compared with control (Sapotsky et al., 2005).

Katoch (2007) at the PR proteins participate a significant responsibility in plant tolerance aligned with pathogens.

The bands presented in 100 μM (SA+BBTMV) and 100 μM SA treatments were greatly accumulated in treatments of tolerance cultivars. The protein subunits were less or more build up depending on SA application.

Due to BBTMV infection, level of 50 and 100 μM (SA+BBTMV) and 100 μM (SA) the protein outline was confirmed novel bands of weights compared to the control in all broad bean leave cultivars.

In the main, the concentration of protein bands of infected lanes was less than that of the control in susceptible cultivars but, more than resistance. SA treatments have a very important role in the accumulation of many PR-proteins responsible for induction of resistance and the presence of a line of defense in plants against virus infection. Loake and Grant (2007) and Radwan et al. (2010) mentioned that the high increase from several novel protein subunit were detected. Infected \been leaves by BYMV observed novel manufactured polypeptide called PRPs in rejoinder to SA application under BYMV infection. In infected tobacco leaves by TMV PRPs were accumulated under SA treatments (White, 1979). Newly, pea leaves samples were collected after treatment with 5 mM salicylic acid (SA) resulted in the appearance of a new protein band of 96.7 kDa 48 h after treatment (Katoch, 2007).

In conclusion, the outcome of the their exploration proposed that the protective action of SA treatments may be associated with a reduction in concentration of virus, infection percentage and severity of BBTMV disease with virus infection. On the other hand, Overall, the BBTMV infection under SA applications caused accumulation of protein contents and founded novel bands of polypeptides in response to infection of BBTMV under SA application in Vicia faba L. plants leaves.

REFERENCES
Bond, D.A., 1987. Recent developments in breeding field beans (Vicia faba L.). Plant Breeding, 99: 1-26.
CrossRef  |  

Chaturvedi, R. and J. Shah, 2007. Salicylic Acid in Plant Disease Resistance. In: Salicylic Acid: A Plant Hormone, Hayat, S. and A. Ahmad (Eds.). Springer, Netherlands, pp: 335-370.

Clark, M.F. and A.N. Adams, 1977. Characteristics of the microplate method of enzyme-linked immunosorbent assay for the detection of plant viruses. J. Gen. Virol., 34: 475-483.
CrossRef  |  PubMed  |  Direct Link  |  

Elbadry, M., R.M. Taha, K.A. Eldougdoug and H. Gamal-Eldin, 2006. Induction of systemic resistance in faba bean (Vicia faba L.) to bean yellow mosaic potyvirus (BYMV) via seed bacterization with plant growth promoting rhizobacteria. J. Plant Dis. Prot., 113: 247-251.
Direct Link  |  

Fraser, R.S.S., 1982. Are pathogenesis-related proteins involved in acquired systemic resistance of tobacco plants to tobacco mosaic virus. J. Gen. Virol., 58: 305-313.
Direct Link  |  

Katoch, R., 2007. Induction of a pathogenesis-related protein in pea after treatment with inducers or inoculation with Erysiphe polygoni. J. Vegetable Sci., 12: 15-25.
CrossRef  |  

Laemmli, U.K., 1970. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature, 227: 680-685.
Direct Link  |  

Loake, G. and M. Grant, 2007. Salicylic acid in plant defence: The players and protagonists. Curr. Opin. Plant Biol., 10: 466-472.
CrossRef  |  

Mwanamwenge, J., S.P. Loss, K.H.M. Siddique and P.S. Cocks, 1998. Growth, seed yield and water use of faba bean (Vicia faba L.) in a short-season Mediterranean-type environment. Anim. Prod. Sci., 38: 171-180.
CrossRef  |  Direct Link  |  

Paul, H.L. and L. Quantz, 1959. Uber den wechsel der konzentration des echten ackerbohnenmosaik-virus in ackerbohnen [About the change of concentration of rights faba bean mosaic virus in field beans]. Archiv fur Mikrobiologie, 32: 312-318.
CrossRef  |  

Quantz, L., 1953. Investigations of a seed via portable mosaic virus of broad bean (Vicia faba). Phytopathol Zeitsch, 20: 421-447.

Radwan, D.E.M., G. Lu, K.A. Fayez and S.Y. Mahmoud, 2008. Protective action of salicylic acid against bean yellow mosaic virus infection in Vicia faba leaves. J. Plant Physiol., 165: 845-857.
CrossRef  |  

Radwan, D.E.M., K.A. Fayez, S. Younis Mahmoud, A. Hamad and G. Lu, 2007. Physiological and metabolic changes of Cucurbita pepo leaves in response to zucchini yellow mosaic virus (ZYMV) infection and salicylic acid treatments. Plant Physiol. Biochem., 45: 480-489.
CrossRef  |  Direct Link  |  

Radwan, D.E.M., K.A. Fayez, S.Y. Mahmoud and G. Lu, 2010. Modifications of antioxidant activity and protein composition of bean leaf due to Bean yellow mosaic virus infection and salicylic acid treatments. Acta Physiologiae Plantarum, 32: 891-904.
CrossRef  |  Direct Link  |  

Sapotsky, M.V., S.A. Romanova, A.M. Polyakova and V.I. Malinovsky, 2005. The correlation between severity of disease symptoms and the accumulation of viral antigen and acidic pathogenesis‐related proteins in the leaves of thorn-apple plants infected with different isolates of potato virus X. J. Phytopathol., 153: 440-444.
CrossRef  |  

Van Loon, L.C. and A. Van Kammen, 1970. Polyacrylamide disk electrophoresis of the soluble leaf proteins from Nicotiana tabacum var. Samsun and Samsun NN III. Influence of temperature and virus strain on changes induced by tobacco mosaic virus. Virology, 40: 199-211.

Van Loon, L.C. and E.A. Van Strien, 1999. The families of pathogenesis-related proteins, their activities and comparative analysis of PR-1 type proteins. Physiol. Mol. Plant Pathol., 55: 85-97.

Van Loon, L.C., 1985. Pathogenesis-related proteins. Plant Mol. Biol., 4: 111-116.
CrossRef  |  Direct Link  |  

Van Loon, L.C., W.S. Pierpoint, T. Boller and V. Conejero, 1994. Recommendations for naming plant pathogenesis-related proteins. Plant Mol. Biol. Rep., 12: 245-264.
CrossRef  |  Direct Link  |  

White, R.F., 1979. Acetylsalicylic acid (aspirin) induces resistance to tobacco mosaic virus in tobacco. Virology, 99: 410-412.
CrossRef  |  PubMed  |  

Yang, X., L. Kang and P. Tien, 1996. Resistance of tomato infected with Cucumber mosaic virus satellite RNA to potato spindle tuber viroid. Ann. Applied Biol., 129: 543-551.
CrossRef  |  Direct Link  |  

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