Serological ELISA Test of Some Black Currant and Red Currant Cultivars

Volume 17(4), 36- 38, 2013 JOURNAL of Horticulture, Forestry and Biotechnology www.journal-hfb.usab-tm.ro

Serological ELISA test of some Black currant and Red currant cultivars

Diaconescu M1*.,Hoza D1., Ion Ligia1

1University of Agriculture Science and Veterinary Medicine Bucharest, Faculty of Horticulture

Corresponding author. Email: [email protected]

Abstract Red and black currants are, for all practical purposes, the same Key words fruit, differing only in color. Currants are a popular fruit among northern Europeans. The fruit is little known in Romania because the plant was blamed anticorps, redcurrant, black for spreading the disease, so for a lot of time was restricted in the culture. currant, Elisa, resistance, Red and white currants are, in fact, relatively resistant to the disease. The virus new researchs in this field, is now renewed interest in Romania in these fruits. The main objectifs for this work is to test some of the bleak and red currant cultivars (Deea, Roxia, Elita 124, Abanos, Triton, Rolan, , JH Van tets, Det van, Tatran, Kzvana) for the two currant’s viruses Blackcurrant reversion virus (BCRV) and Rasberry leaf curl luteovirus (RLCV) using serological Elisa test . After preliminary tests two romanian cultivars Roxia and Elita 124 was foundes negatifs in Elisa test ant coul be an imporatnt promise for the next molecular determination.This paperpresents data from PhD thesis part of the project POSDRU/107/1.5/S/76888, funded by European Social Fundthroughthe Sectoral Operational ProgrammeHuman Resources Development 2007-2013.

The plant genus Ribes includes many types of fruits, production levels. The cultivated blackcurrant (R. including the blackcurrant and red currant. The nigrum L.) is grown throughout temperate areas of blackcurrant plant is a flowering shrub that thrives in Europe and New Zealand. These fruits are mainly used temperate regions and is suitable for cultivation on by the processing industry for concentrate and juice small farms. Romania, blackcurrant are widely production as well as for deep freezing as individually produced because production costs are low, the plants quick frozen (IQF) fruit. They have high nutritional are hardy and easily managed, and little pest control is and health values for people because of the high level required [1]; [2]. However, several pests can affect of ascorbic acid (vitamin C) and anthocyanins [4]; [5]; blackcurrant production in this region [16] [3]. Recently, blackcurrants have also appeared on the EulechriopsrubiHespenheide, 2005 (Coleoptera: fresh market as dessertfruit due to the perceived health Curculionidae), Anastrephafraterculus[15] (Diptera: benefits associated with high antioxidant capacity and Tephritidae), leaf beetles (Coleoptera: polyphenolic content [8], [13] ,[11]; [14]. Chrysomelidae), leafcutter ants (Hymenoptera: Formicidae), and mites (Acari). In addition, the leaf- Materials and Methods eating larvae of Herpetogramma bipunctalis [17] (Lepidoptera: Crambidae), commonly known in our Plant material evaluated were included, 10 red and country. In 2010, the population size of H.bipunctalis bleak currant cultivars Deea, Roxia, Elita 124, Abanos, in the state was the largest in March and April, and Triton, Rolan, , JH Van tets, Det van, Tatran, Kzvana. smallest in November [6]. Larvae of H.bipunctalis For large-scale indexing of viruses , yet most widely cause damage by feeding on blackcurrant leaves, which used with good results is DAS - ELISA standard - curl as a form of protection, resulting in early Double Antibody Sandwich double- antibody sandwich senescence (leaf drop) (Nava, DE. Personal . (Fig 1 and 2) communication). [12]. The main production The principle of the method is that the antibody problem,mostly in blackcurrants, are connected with sandwich component - antigen - antibody conjugate - the spread of the gall mite and blackcurrant reversion specific enzyme hydrolyzes the substrate and thus its virus (BRV) due to lack of effective chemical or color changes ( antigen is bound to the antibody set and culturalcontrols for this pest. Recently, several Coated polystyrene plates ) highlighting realizing cultivars resistant to the pest and BRV have been itself. The color reaction can be detected by means of a released from breeding programs in different countries. photometer , the intensity of which is proportional to Among these crops the blackcurrants are the most virus concentration . The sample in question can be important due to large areas in cultivation and high considered infected if the value exceeds the average

36 extinction healthy controls twice. For serological different viruses or virus strains ). Variant TAS / DASI diagnosis can be used both polyclonal antisera and - ELISA enables serological differentiation of PPV monoclonal universal . To identify viral strainsusing isolates using monoclonal antisera specific targeting specific monoclonal antisera . DAS - ELISA is different parts of the capsid protein (specific each sometimes used with smallchanges in the enzymes to strain . The method allows the testing of large-scale make the test more economical. SometimesDAS high ELISA to detect viruses fruit trees which are available specificity - the direct ELISA problematic and is polyclonal antisera and / or monoclonal antibodies therefore preferred indirect ELISA . This technique is However, the method has limitations such as the fact also used to determine the serological relationships that some viruses may exist in very low concentrations between viruses (same conjugate can be used for in trees or may have irregular distribution.

Fig 1 and 2. Aspects from Elisa tests.

Results and Discussions serologic testing currant varieties under study in virus reversiunii currant (BCRV) and leaf curling virus DAS-ELISA test is most often used in the diagnosis of (RLCV). This demonstrates that not all varieties but viruses. However, serological techniques that can field experience symptoms when tested serologically detect low levels of virus, are suitable for the detection viral infection is emphasized, as is the case of varieties of viruses symptoms. Table 1 presented results on Roxie, Triton and Rolan (red currant).

Table 1 Results concerning the serological tests Elisa fromBlackcurrant reversion virus (BCRV) and Rasberry leaf curl luteovirus( RLCV) BCRV RLCV DAS-ELISA DAS-ELISA Variety Number of probs Intensitaty of Intensity of (DO= 405nm) (DO= 405nm) simptoms simptoms Deea 20 - - - - Roxia 20 - + + + Elita 124 20 - - - - Abanos 20 + ++ + + Triton 20 - ++ - + Rolan CR 20 - + + + JH van Tets 20 ++ ++ + + Det Van 20 + + + + Tatran 20 - +++ + + Kzvana 20 - + + +

Interestingly, among the 10 varieties studied this aspect The assumptions for resistance to viral genetic control of resistance to viruses have revealed two elite varieties currant the species referred to by different authors [7], Deea and Elita124 were not detected positive Elisa [9], [10] consider that it is the dominant resistance (Figures 3 and 4) Which would could open up new allele. avenues of research and molecular analyzes.

37 Infection process was different for each individual, with different degrees of infection.

Fig.3 and 4. Results of Elisa test reaction

Conclusions References

This shows that not all varieties show the symptoms in 1.Antunes, L., 2002. Amora-preta: Nova opção de the field condicions after when was tested cultivo no Brasil. Ciência Rural, vol. 32, no. 1, p. 151- serologically and viral infection is emphasized, as is 158. the case of varieties Roxie, Triton and Rolan (red 2. Antunes, L., Gonçalves, Ed. and Trevisan, R., 2010. currant). Fenologia e produção de cultivares de amoreira-preta Among the 10 varieties studied this aspect of resistance em sistema agroecológico. Ciência Rural, vol. 40, no. to viruses have revealed two elite varieties Deea and 9, p. 1929-1923. Elita 124 were not detected positive Elisa (Figures 3 3. Brennan, R.M., Jorgensen, L., Hackett, C., and 4) Which could open new research perspectives Woodhead, M., Gordon, S. L. and Russell, J. 2008b. and molecular analyzes The development of a genetic linkage map of Gene hypothesis for the control of viral resistance to blackcurrant (Ribes nigrum L.) and the identification of the species to which the currant different authors [7], regions associated with key fruit quality and agronomic [9], [10] consider that it is the dominant resistance traits. Euphytica 161:19-34. allele. 4. Banaszczyk, J. and Pluta, S. 1997. Quality characteristics of fruits of new blackcurrant selections Acknowledgement released at Skierniewice. Journal of Fruit and Ornamental Plant Research 5:13-17. This paper presents data from Phd thesis part of the 5. Brennan, R.M. 2008. Currants and gooseberries. project POSDRU/107/1.5/S/76888, funded by p.177-196. In: J.F. Hancock (ed.), Temperate fruit crop European Social Fund through the Sectoral Operational breeding. Springer, Berlin. Programme Human Resources Development 2007- 6. Crozier, A. 2002. Small fruit as sources of dietary 2013. antioxidants. 2002. Atca Hort.585:459-465.