Project ERA 226 “Capsicum Balkan Biodiversity”

Welcome to ERA 226 project

Capsicum Balkan biodiversity

Exploration of Balkan biodiversity of Capsicum spp. to extract
biotic stress resistant germplasm

Pepper (Capsicum annuum var. annuum L.) is one of the most important and wide spread vegetables in the Balkan region. A great number of local populations with specific characteristics for shape, colour, taste, biological value and type of use are grown in different Balkan countries together with the main commercial cultivars. Partially the old native forms are not within attention of the sort maintenance selection, but they are preserved in limited area and are valuable genetic resources. In 2010 ERA 226 project started in the framework of the international program SEE-ERA.NET PLUS setting pepper as target species, with two main tasks:

1) to collect, describe and explore the existing Capsicum biodiversity in the partner countries (Albania, Bulgaria, FYR Macedonia, Greece and Serbia) related to valuable traits;

2) to update knowledge about the most economically important and emerging pathogens on Capsicum in the Balkan region and to form pathogen collection.

PROJECT PARTNERS

Sub-project Institution Team member
ERA 226/01
Project coordinator
Institute of Plant Physiology and Genetics – Bulgarian Academy of Sciences, Acad. G. Bonchev Str., Bl. 21, 1113 Sofia, Bulgaria Dr. R. Rodeva
Dr. R. Pandeva
MSc. Z. Stoyanova
MSc. M. Petkova
ERA 226/02 Agricultural University of Tirana – Tirana, Albania Prof. J. Merkuri
Dr. M. Cara
Dr. E. Tome
MSc. V. Vladi
ERA 226/03 Institute of Plant Genetics Resources – Sadovo, Bulgaria Prof. L. Krasteva
Dr. S. Neykov
Dr. K. Uzundzhalieva
Dr. P. Chavdarov
Assist. N. Velcheva
ERA 226/04 “Maritsa” Vegetable Crops Research Institute – Plovdiv, Bulgaria Dr. D. Kostova
Assist. G. Pasev
Dr. V. Todorova
ERA 226/05 Goce Delchev University – Stip, FYR Macedonia Prof. I. Karov
Prov. S. Mitrev
Prof. V. Ilieva
Assist. B. Kovacevik
ERA 226/06 Plant Protection Institute of Patras – NAGREF, Greece Dr. I. Manoussopoulos
MSc. I. Lagogiannis
MSc. S. Goudoudaki
ERA 226/06 Institute for Vegetable Crops – Smederevska Palanka, Serbia Dr. Dejan Cvikic
Dr. M. Mijatovic

WORKSHOPS

The kick-off meeting (28.11-03.12.2010) organized by the Institute of Plant Genetic Resources was carried out in Plovdiv and in Sadovo, Bulgaria. During the meeting the basic methods for expedition organization, trip itinerary and duration and also for the description, maintenance and reproduction of the collected material were defined. It was combined with two joint collecting tours in Southeastern and Central Bulgaria.
The workshop in Macedonia (23-27.08.2011) took place in the Goce Delcev University – Shtip, combined with three collection trips.
The workshop in Serbia (11-15.09.2011) was carried out in the Institute for Vegetables Crops – Smederevska Palanka, and included extensive expedition program with visits of pepper producers.
The workshop in Albania (13-16.10.2011) was combined with participation in 5th Balkan Symposium of Vegetables and Potatoes (09-12.10.2011) in Tirana, where ERA 226 project was presented with two reports. The main program was accomplished in the Agricultural University of Tirana, the Laboratory of Plant Protection in Durres and the Agricultural Centre for Technology Transfer in Lushnja. The collecting missions included visits of open and protected fields in different parts of the country.
The workshop in Greece (25-29.06.2012) was also combined with expedition trips to pepper producers.
Final meeting of ERA 226 was carried out in Sofia (12-15.09.2012). Participants of every partner institution presented the main results obtained in the framework of the project. International pepper collection was organized as a part of the scientific program of the meeting. It was arranged at the experimental field of IPPG in 2012 as a comparative demonstrative trial, in which most of pepper specimens, collected from the partner countries, were grown under uniform conditions.

MAIN RESULTS

Task 1. Collection and description of local Capsicum specimens (domestic accessions, varieties, breeding lines) typical for Balkan region. Pepper collection created during the implementation of ERA 226 project included 235 local forms, situated in Albania (17), Bulgaria (134), Greece (34), Macedonia (11) and Serbia (39). Morphological and phenological observations as well as biometric measurements on the basis of 67 features were conducted according to Descriptors for Capsicum with some additions. Significant diversity of local pepper forms was established in the collected material. Fruit shape was used as a major trait for its arrangement, with 5 variants: elongated, almost round, triangular, campanulate and blocky. Two additional features, fruit length and pungency were also applied for more precise grouping of this material. Peppers with elongated fruits composed the most variable group including different types known in Bulgaria as ‘shipka’, ‘ribki’ (small fishes), ‘roga’ (horns) and ‘kapiya’. Most of them are also popular in Macedonia, Serbia and Albania. Second group was more uniform, consisting of two main types, small-fruited pungent almost round ‘cherries’ and large-fruited sweet flat-round ‘ratund’ (kambi). The group of triangular peppers was the most numerous in our study. It consists of predominantly sweet, conical or two-side pressed large-fruited forms named ‘kapiya’, very popular оn the Balkans. They were well presented in all collections. Small-fruited hot representatives of this group are also known as ‘shipka’. Campanulate-fruit type was very feebly overlapped with two Serbian accessions. The participation of blocky type peppers, named ‘dolma’ was relatively small. They were uniformly situated in all collections except Macedonian. All ERA 226 local representatives belong to C. annuum and are annual in their life cycle. Plant, stem and leaf characteristics of the main accession groups showed relatively stable uniformity in significant part of the studied traits or fluctuations between consecutive grades. More variable were characters as plant habit, tillering and leaf density. Flowers with all types of positions were presented in ERA 226 pepper collections. The main colour of unripe fruits was green. Yellow fruits had some Albanian, Bulgarian and Serbian accessions. The mature fruits were predominantly red.

Task 2. Survey of naturally occurring diseases affecting pepper in participating countries. During the field trips in different countries plant parts with disease symptoms were collected and preserved. Virus detection was based on serological methods using virus specific antibodies. The standard method was ELISA although dot-blot ELISA was also used in some cases. The attention was focused on those species that infected C. annuum naturally, which have been reported to occur in the Balkan region in high frequency. There were seven such viruses, namely: Pepper mild mottle virus (PMMV), Tobacco mosaic virus (TMV), Tomato mosaic virus (ToMV), Potato virus Y (PVY), Cucumber mosaic virus (CMV), Alfalfa mosaic virus (AMV), and Tomato spotted wilt virus (TSWV). Specific genus antibodies for two most numerous genera, namely, Potyvirus and Tobamovirus covering a total of 13 viruses (9 Potyvirus and 4 Tobamovirus) were used where possible. The most important viruses, according to these data were CMV and PVY although with different importance among different countries. Indeed, CMV seemed to be more frequent in pepper crops in the northern countries (Bulgaria, FYROM, Serbia and Albania) than in south-western Greece. On the other hand, PVY was more frequent in Greek crops and absent in all northern countries except Serbia. TMV and ToMV were sporadically found in crops in Albania. Very important observation of this work was the occurrence of a high number of plants with virus-like symptoms free of any of the tested viruses. This could be due to environmental stress or other circumstances (irrigation, pesticide usage etc.) that affects plants, making them to show virus-like symptoms. At the same time, it is quite plausible that another important virus may be present, having not been detected by our approaches.

The investigation of the bacterial disease showed the presence of three main plant pathogenic bacteria: Erwinia carotovora ssp. carotovora, Pseudomonas syringae pv. syringae and the most spread one Xanthomonas campestris pv. vesicatoria causing bacterial soft rot, bacterial leaf spot and bacterial spot, respectively. Bacteral diseases were present but did not cause significant yield loss except in the nursery production. The total number of the examined isolates was estimated at around 444 from which 214 were identified as pathogenic.

Fungal diseases affecting root/stem base were the most frequently encountered problem for pepper crop damaging the plants at any stage. Among them the most prevalent were Rhizoctonia solani, Fusarium solani, F. oxysporum f.sp. capsici and Verticillium dahliae. Lately, Macrophomina phaseolina and Colletotrichum coccodes were isolated from the roots of wilted plants with increasing frequency separately or in mixture with other soil fungi. Phytophthora capsici was very important pathogen in the past but during the study it was isolated mostly from seedlings in the nursery. In 2012, heavily infected by Phytophthora spp. plants were observed in the demonstrative field experiment in Sofia. It was established that the isolates from diseased pepper plants and from the soil belong to P. capsici and to P. cryptogea, respectively. Alternaria alternata, A. solani, Fusarium spp. and Botrytis cinerea were commonly isolated from pepper fruits. Sclerotinia sclerotiorum was mainly recorded in protected fields. Powdery mildew incited by Leveillula taurica was often found when greenhouse growing was combined with high relative humidity. In the open fields this disease was established mainly in the end of growing season. As an exception it was found the most spread fungal pathogen in Albania and Macedonia during the studied period. Special emphasis deserved new and emerging fungal pathogens. Three Colletotrichum species (C. coccodes, C. acutatum and C. gloeosporioides) were identified in Bulgaria. C. coccodes was the most prevalent in Bulgaria but in Macedonia was found for the first time. A comparative morphological, cultural and molecular characterization was made of C. coccodes isolates obtained from pepper fruits and roots in both countries. Two PCR primer sets were used for sequencing the ribosomal internal transcribed spacer (ITS1 and ITS2) regions. Single products of ~450 bp and ~350bp were amplified by the genus-specific (Cc1F1/Cc2R1) and the species-specific primers (Cc1NF1/Cc2NR1), respectively. One unusual Colletotrichum sp. was found on naturally infected fruits of pepper in Bulgaria and Serbia. A single band of about 470 bp in length was detected when C. gloeosporioides-specific primer set CgInt/Its4 was used suggesting that the unusual species might belong to the C. gloeosporioides group. Another fruit disease caused by Phomopsis capsici was recently reported in Bulgaria. In 2011 the fungus was established in Macedonia for the first time. Several new causal agents were found as Phoma sp. on pepper fruits (Albania, Macedonia) and Ascochyta sp. on leaves (Bulgaria, Macedonia), which remained unidentified to the species level and their taxonomical status will be delimitated in the near future.

Task 3. Project management. The current project was a good opportunity to activate the mutual contacts and to start the first step for collaborative research programs – joint expeditions, meetings to discuss problems, taking decisions for future, etc. This regional collaboration contributed for dissemination of good practice across partner institutes/universities and knowledge transfer. Without diminishing the freedom of individual scientists, efforts were made to create synergism through positive interaction, resulting in considerable added value in scientific terms. The enhancement that participants achieved (to knowledge, skills abilities and other attributes) as a result of their experience during the project implementation was also a substantial added value. The involvement of young scientists in the action plan contributed for improvement of their expertise and capability to manage the assessment of genetic resources and pest identification and characterization. Two of them are in process of PhD thesis preparation.

DISSEMINATION OF KNOWLEDGE TO THE KEY AUDIENCES

Participation in scientific events:

Dissemination of knowledge to the key audiences and publications:

  1. Krasteva, L., R. Pandeva, R. Rodeva, V. Todorova, S. Neykov, K. Uzundzhalieva, N. Velcheva, D. Cvikic, E. Tome, V. Ilieva. 2012. Pepper as a target object of SEE-ERA.NET project. Acta Horticulturae (ISHS) 960: 151-158.
  2. Rodeva, R., D. Kostova, P. Chavdarov, M. Mijatovic, J. Merkuri, M. Cara, G. Pasev, Z. Stoyanova, I. Karov, S. Mitrev, B. Kovacevik, S. Goudoudaki, I. Manoussopoulos. 2012. Pepper diseases in Balkan region. Acta Horticulturae (ISHS) 960: 365-370.
  3. Rodeva, R., Z. Stoyanova, E. Surviliene, P. Chavdarov. Fungi associated with seed and plant infection of pepper. 18th Symposium of the Baltic Mycologists and Lichenologists, Dubingiai, Lithuania, 19-22.09.2011.
  4. Rodeva, R., I. Karov, Z. Stoyanova, B. Kovacevik, V. Manova, R. Georgieva. 2012. Phomopsis capsici and Colletotrichum coccodes infecting pepper in Macedonia. International Symposium: Current Trends in Plant Protection. Proceedings. Institute for Plant Protection and Environment, Belgrade, Serbia, 257-263.
  5. Stoyanova, Z., R. Rodeva, V. Manova, L. Stoilov, M. Mijatovic. 2012. Unusual Colletotrichum sp. associated with pepper fruit anthracnose in Bulgaria and Serbia – Preliminary results. International Symposium: Current Trends in Plant Protection. Proceedings. Institute for Plant Protection and Environment, Belgrade, Serbia, 299-306.
  6. Cvikic, D., L. Krasteva, R. Pandeva, R. Rodeva, V. Todorova, S. Neykov, N. Velcheva. 2012. Genetic resources of pepper in Serbia – SEE ERA NET project. Book of Abstracts. International Scientific Conference “130 Years Agricultural Science in Sadovo”, Sadovo, Bulgaria, 5-6 June 2012.
  7. Karov, I., B. Kovacevic, S. Mitrev, R. Rodeva, Z. Stoyanova, P. Chavdarov. Fungal diseases of pepper in Eastern Macedonia in 2011-2012. 37th International Symposium for Plant Protection in Ohrid, Macedonia, 20-21.12.2012. Plant Protection (Skopje) (in press).
  8. Stoyanova, Z., R. Rodeva, I. Karov, B. Kovacevik, V. Manova, R. Georgieva. Morphological and molecular characterization of Colletotrichum coccodes isolated from pepper cultivated in Bulgaria and Macedonia. J. Nat. Sci., Matica Srpska Novi Sad, 124, 249-262.

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