Slovene Plant Gene Bank (SPGB)

and Genetic Resources Programme

Second Meeting of the ECPGR Working Group on Leafy Vegetables 8 – 9 October, Ljubljana, Slovenia

Vladimir MEGLIČ, Jelka ŠUŠTAR VOZLIČ

Early projects to collect Slovenian autochthonous populations, ecotypes and landraces of agricultural species were initiated about 60 years ago.

In 1996 the Ministry of Agriculture, Forestry and Food started financing the Slovene Plant Genetic Resources Programme with the goal to maintain, evaluate, regenerate and preserve Slovenian autochthonous species, ecotypes, populations and landraces of agricultural, medicinal and aromatic plants. Slovene Plant Gene Bank (SPGB) – Gene Bank of Agricultural Crop Species was established.

SPGB includes Slovenian varieties, old varieties, landraces, various populations, clones and lines bred from autochthonous plants and ecotypes from the natural habitat important for food and agriculture.

Slovene Plant Gene Bank

ECPGR Leafy Vegetables WG Meeting, 8-9 October 2013, Ljubljana

SLOVENE PLANT

GENE BANK Central Seed Bank

(AIS LJ)

PSARS (MAE) - Central Database

MEDPLANT Database

Faculty of Agriculture and Life Sciences University of Maribor

NGO Urban Furrows, botanical gardens, schools, …

Agricultural Institute of Slovenia, Ljubljana

Slovenian Institute for Hop Research and and Brewing, Žalec

Biotechnical Faculty University of Ljubljana

Slovene Plant Gene Bank

ECPGR Leafy Vegetables WG Meeting, 8-9 October 2013, Ljubljana

Institution No. of accessions

Biotechnical faculty >1200

Agricultural Institute of Slovenia

>3200

Slovenian institute for Hop Res.

93

Faculty of agriculture and Life Sciences

>300

Total >6000

Biotechnical faculty, Agronomy Dept., University of Ljubljana Fagopyrum, Zea mays, fruit trees, forage crops, medicinal and aromatic plants.

Agricultural Institute of Slovenia, Ljubljana Grain legumes, Allium, Solanum, Triticum, Brassica, Lactuca, forage and fodder crops, Rubus, Vitis,….

Slovenian Institute for Hop Research and Brewing Humulus, medicinal and aromatic plants.

Faculty for Agriculture and Life Sciences, University of Maribor Fruit trees, Vitis

Responsability

ECPGR Leafy Vegetables WG Meeting, 8-9 October 2013, Ljubljana

Slovene Plant Gene Bank

• Collecting

• Documentation

• Multiplication

• Conservation

• Storage

ECPGR Leafy Vegetables WG Meeting, 8-9 October 2013, Ljubljana

-Wild relatives of cultivated crops - Landraces and populations - Old cultivars

Collecting

ECPGR Leafy Vegetables WG Meeting, 8-9 October 2013, Ljubljana

- Expeditions throghout Slovenia - Bilateral and multilateral projects - Donations and exchanges - Schools, adds, ….

Collecting

ECPGR Leafy Vegetables WG Meeting, 8-9 October 2013, Ljubljana

Dependant on crop species:

-field,

-screenhouses,

-greenhouses,

-in vitro

Multiplication

ECPGR Leafy Vegetables WG Meeting, 8-9 October 2013, Ljubljana

Characterisation and evaluation: -Chemical analysis of 38 ecotypes of Dactylis glomerata L.

-Characterisation of 21 red clover ecotypes (morpf., quality (NIR)).

-Characterisation of 17 Medicago falcata ecotypes (morpf., quality (NIR)).

70

80

90

100

110

120

130

140

270 290 310 330 350 370 390

vsebnost SVL (g kg-1

SS)

vse

bn

ost

SB

(g

kg

-1S

S)

Characterization and evaluation

ECPGR Leafy Vegetables WG Meeting, 8-9 October 2013, Ljubljana

- Germination - Moisture content - Seed purity

Conservation – seed quality

ECPGR Leafy Vegetables WG Meeting, 8-9 October 2013, Ljubljana

Working collection (mid-term storage): - 500 - 1000 seeds/accession stored at + 4 C.

Basic collection (long-term storage at –20°C): -located at the Agricultural Institute of Slovenia.

Accessions are also kept in ‘in vitro’ conditions and ‘in vivo’ in permanent plantations for hops, fruit, grapevine and some species of medicinal and aromatic plants.

Storage

ECPGR Leafy Vegetables WG Meeting, 8-9 October 2013, Ljubljana

-Research: - Diversity studies, - Genetic erosion, - Population genetics, - Mapping, - Conservation and preservation studies….

-Breeding, -Immediate use on farm…

Use of plant genetic resources

ECPGR Leafy Vegetables WG Meeting, 8-9 October 2013, Ljubljana

Genetic Resources Programme at the Agricultural Institute of Slovenia

Group of species Species Number of accessions

Grain legumes Phaseolus spp. 1125

Vicia faba 37

Skupaj 1162

Fodder crops Trifolium sp. 199

Medicago sp. 39

Vicia sp. 69

Lotus sp. 33

Lolium sp. 30

Festuca sp. 83

Dactylis 110

Phleum 39

Travniške zeli 65

others 202

Skupaj 869

Field crops Zea mays 11

Triticum 76

Secale 12

Papaver 26

Panicum 15

ostale 60

Skupaj 200

Vegetables Lactuca sp. 227

Allium cepa 56

Brassica oleracea 11

Diplotaxis spp. 18

ostale(Cichorium, Valerianella, Solanum Lyc.) 14

Allium sativum 8

Skupaj 334

Small fruites Vaccinium corymbosum, Rubus , Ribes, Fragaria 230

Apple trees M. domestica, M. floribunda 256

Grapewine Vitis 90

Potato Solanum tuberosum L. 34

Other potato species Solanum sp. 18 + 14 diploid populations

Total 642

Total 3207

ECPGR Leafy Vegetables WG Meeting, 8-9 October 2013, Ljubljana

Collection of Phaseolus accessions

1081 accessions (1020 Slovenian, 61 from other GB)

– 841 (Phaseolus vulgaris L. ssp. vulgaris var. vulgaris),

– 187 (Phaseolus vulgaris L. ssp. vulgaris var. nanus Aschers.),

– 53 (Phaseolus coccineus L.)

Multicrop passport podatki – available for all accessions

Data on characterization and evaluation – available for a

limited number of accessions

ECPGR Leafy Vegetables WG Meeting, 8-9 October 2013, Ljubljana

Morphological markers Biochemical markers (phaseolin) Molecular markers (RAPD, AFLP, SSR)

Disease resistance (Anthracnose)

Drought resistance (proteases, gene expression, psyiological analysis, proteomics)

Evaluation of common bean accessions

ECPGR Leafy Vegetables WG Meeting, 8-9 October 2013, Ljubljana

Drought resistance - gene expression

KAVAR T, MARAS M, KIDRIČ M, ŠUŠTAR VOZLIČ J, MEGLIČ V. 2008. Mol. breed. 21: 159-172.

• Changes in gene expression in the leaves at different levels of dehydration were identified by DDRT PCR and confirmed by quantitative real-time PCR.

• The levels of 15 transcripts were changed significantly in all cultivars, 8 being increased and seven decreased. At least six transcripts corresponded to those whose expression is altered under drought in Arabidopsis thaliana. Five of the genes identified have not previously been reported as being influenced by drought. Only minor differences in expression were found between the different cultivars.

ECPGR Leafy Vegetables WG Meeting, 8-9 October 2013, Ljubljana

Drought resistance - proteomic approach

The aim is to identify proteins differentially expressed due to water deficit stress (Starozagorski čern – sensitive, Tiber – tolerant).

Proteins were extracted from stressed and control plants, labelled with different fluorescent dyes and were separated using two-dimensional gel electrophoresis (2-DE). Protein spots that were significantly changed in abundance were excised, trypsin digested and analysed on LC-MS/MS

ECPGR Leafy Vegetables WG Meeting, 8-9 October 2013, Ljubljana

Drought resistance - proteomic approach

Analysis of functional network. Two clusters of highly interacting protein nodes represents proteins involved in photosynthesis and energy metabolism.

2DGelectrophoresis of leaf proteins from Starozagorski (differentially expressed and identified protein spots).

Identified proteins can be divided in different functional categories that include metabolic proteins, cellular defense/stress proteins, proteins involved in reactions of photosynthesis and proteins with unknown function

ECPGR Leafy Vegetables WG Meeting, 8-9 October 2013, Ljubljana

Thank you for your attention!