Nordic/ECPGR Joint Workshop, 19-21 September 2016, Vilnius, Lithuania

CWR CONSERVATION STRATEGIES Combining in situ and ex situ CWR conservation

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Miguel Ângelo Almeida Pinheiro de Carvalho miguel.carvalho@staff.uma.pt

http://www.uma.pt/isoplexis

Plant genetic resources for food security and ecosystem services

Planning and implementing national and

regional conservation strategies

2 http://www.uma.pt/isoplexis

Geographical Madeira Region, is an archipelago localized in Atlantic Ocean

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Combining active in Situ and ex Situ CWR Conservation Main outcome of one of AEGRO project workpackages has the study of Beta patula distribution and population baseline and the design of a plan to setup a genetic reserve that could define the most appropriate site for conservation of plant genetic diversity. The plan of genetic reserve has proposed to Madeiran IFCN and submitted as proposal to the Life program for funding. And, as a result project LIFE12.NAT.PT.000195 has approved and is currently underway with goals to: a) restore the native ecosystems in the protected areas of Ponta de São Lourenço and

Desertas Islands. b) And, implement the genetic reserve for Beta patula and other beet CWRs.

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Beta patula Aiton

Copyright: University of Madeira Copyright: University of Madeira

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Project LIFE12.NAT.PT.000195 Actions to be undertaken to implement the genetic reserve and achieve the in situ conservation of Beta patula. The main objectives of these actions are: a) Understand the ecogeographic and ecological conditions of Beta patula occurrence; a) Establish the Beta patula population sizes and its dynamics; b) Create ex situ collections of Beta patula, that could be used as populations backup or to

supply germplasm for interested parties; c) Analyze the species genetic diversity and its spatial display; d) Delimit the genetic reserve boundaries; e) Evaluate the in situ conservation actions that need to be implemented; f) Establish a middle term management protocol for Beta patula and other CWRs. Project have an execution period of 5 years and Beta population will be monitored in an yearly bases

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Ponta de São Lourenço and Desertas Islands Status protection: Natura 2000 Network sites ID (Partial Protected Area) IC (Full Protected Area)

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Introductory site description Beta patula occurs in Ilhéu Desembarcadouro, Ponta de Lourenço and Ilhéu Chão, Desertas. But for implementation of the genetic reserve for its in situ conservation has proposed Desembarcadouro, Ponta de São Lourenço, based on data generated by AEGRO project. In this site Beta patula showed a wider distribution and highest population sizes, with better chances to ensuring species viable population sizes. Other factors Ponta de São Lourenço shows also high plant diversity occurring 176 plant species: 35 endemism's; 110 native species; 31 introduced species Among them 58 are CWRs.

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Understanding the ecogeographic and ecological conditions of Beta patula occurrence; Ecogeographic data Plant ecogeographic distribution with georefenciation. Habitat description Altitude Soil type Climate and metrological data Map shows the pluviometry data (series of 20 years), showing that species preferences are dry conditions.

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Understanding the ecogeographic and ecological conditions of Beta patula occurrence Ecogeographic data Plant ecogeographic distribution with georefenciation. Habitat description Altitude Soil type Climate and metrological data Map shows the temperature data (series of 20 years), showing that species preferences are relative high temperature and radiation conditions.

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Understanding the ecogeographic and ecological conditions of Beta patula occurrence Ecogeographic data Plant ecogeographic distribution with georefenciation. Habitat description Altitude Soil type Climate and metrological data. Map shows the soil type and CWR distribution, showing that species colonize different soil Types (3 at least).

Quadrado 1

Q 1 32 44 095 016 40 583 19 m 1 2 3 4 Espécie N8 7 6 5 Beta patula 5 40

9 10 11 12 Suaeda vera 1 20

16 15 14 13 Mesembriathemum crystalinum 1 12

Senecio incrassatus 1 3

Sonchus oleraceus 3 3

78 %

Quadrado 5

Espécie NBeta patula 4 32

Suaeda vera 2 16

Mesembriathemum crystalinum 1 2

Crepis divaricata 3 3

53 %

Quadrado 10

Espécie NBeta patula 2 20

Suaeda vera 2 20

Mesembriathemum crystalinum 1 1

Phalaris maderensis 1 1

Sonchus oleraceus 3 2

Medicago serrata 5 6

50 %

Quadrado 13

Espécie NBeta patula 4 20 *

Suaeda vera 1 8

Mesembriathemum crystalinum 4 8 *

Senecio incrassatus 2 2 *

38 %

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Establishing the Beta patula population sizes and dynamics. Beta patula population census methodology Twelve quadrants with 16m2 divided in 1m2 squares, displayed along 3 parallel transepts defined in Desembarcadouro. In each quadrant the species occurrence and abundance has assessed, and de number of Beta patula specimens accounted in 4 randomly selected square meters inside each quadrant. Right quadrants give an idea of species frequencies and quadrants coverage in the field.

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Establishing the Beta patula population sizes and dynamics Population distribution Netto area – 105,000 m2

Real area – < 80,000 m2*

* - under validation.

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Establishing the Beta patula population sizes and dynamics. This methodology allows us to determine the species richness and diversity in the sites/patches of occurrence of Beta patula.

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Establishing the Beta patula population sizes and dynamics. In these figures are summarized data of species occurrence and richness in 2 years population census. Besides 12 other CWRs occurs and share the space with Beta patula.

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diversity indices Q1 Q2 Q3 Q4 Q5 Q6 Q7 Q8

H' 2.689 2.108 2.588 2.362 2.412 0.355 1.907 1.485

E' 0.841 0.690 0.829 0.802 0.742 0.150 0.795 0.473

D 0.184 0.289 0.189 0.242 0.221 0.889 0.286 0.493

diversity indices Q9 Q10 Q11 Q12 Q13 Q14 Q15 -

H' 1.727 2.943 2.258 2.499 0.972 1.914 1.954 -

E' 0.708 0.861 0.598 0.751 0.383 0.590 0.825 -

D 0.350 0.144 0.278 0.214 0.684 0.346 0.315 -

H' = Shannon-Wienner diversity index (ecosystems’ condition); E' = Corrected Eveness; D = Simpson Index (Inverted).

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Establishing the Beta patula population sizes and dynamics

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Establishing the Beta patula population sizes and dynamics. Field population data Beta patula census show the following evolution of knowledge about the plant population sizes:

Beta patula, census Estimative Nº specimens Observations

Early data < 50 Directive of Habitats

Preliminary data 5,000 AEGRO Project

Current census 5,000 to 50,000 Under validation

Our results shows that in actual conditions Beta patula effectives and population sizes are apparently quite consistent and stable. Despite the existence of identified potential threats.

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Establishing the Beta patula population sizes and dynamics. Assessing the role of soil seeds bank in population effectives replacement The size of soil seed bank has determined by sampling soil in 15 square meters established in the neighbors of main quadrants. Soil seed samples has obtained by suction, cleaning and seeds account. Ex situ and in situ germination test has been Realized to determine the seeds viability.

data

altitude,

m

nº

seeds

weight, 100

seeds g

weight, 1,000

seeds g

average 51 2,272 1.98 16.41

max 93 5,073 4.31 31.43

min 19 351 1.19 11.30

Germination

viability, % 10 to 15 (0.8%)

500 seeds

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Creating ex situ collections of Beta patula. These collections have two purposes: a) Create a backup for species conservation purposes that can be use in case of plant reintroduction in nature. b) Obtain germplasm that can be supplied for interested parties and stakeholders. Beta patula populations has been partially sampled along its gradient distribution generating 19 germplasm accessions 15 seed samples was collected in the Desembarcadouro. 4 seed samples was collected in the Ilhéu Chão. Beta patula accessions has been integrated in ISOPlexis Genebank collection (duplicates will be stored in BGM and JKI)

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Managing of the Beta patula germplasm Accession documentation consist on: i) Passport data (accession number, collection number, vernacular name, taxonomic

identification, collector, data of storage); ii) Harvest record (collection site, georeferenciation, site description, soil type, habitat

description, species or samples specific traits, value, threats, data of sampling); iii) Additional accession data (sample size, number of seeds, 100 or 1000 seeds weight,

sanitation and moisture %, viability data); iv) Characterization and evaluation (morph-agronomic, morphometric, phenological,

biochemical or molecular). In populations’ or accessions’ characterization usually IPGRI or species adapted descriptor are used. This information is registered and stored in mechanographical, computer files or in databases (ISOPex and Grin Global).

22 Proceeds certified under the standards 9001:2000 e 14001:2004

ISOPlexis Genebank (Banco de Germoplasma ISOPlexis)

The genebank maintain ex situ and in situ collections of crop (mainly) and priority CWR genetic

resources occurring in Madeira.

Documentation and Information System

Main Germplasm Collections

• Base collection for long time conservation.

• Active collection for research and farmers support.

• Breeding collection with improved varieties and lines.

• Field collection of tropical roots corps (sweet potato, taro, vegetative propagated species, etc.).

Genebank database (ISOPex)

Includes records of 4,308 (2,889) accessions (access 4 of April 2016).

http://www.uma.pt/isoplexis

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Active ex situ Conservation of CWRs BG ISOPlexis (ISOPlexis Genebank) holds 445 accessions of 82 CWR species. The most representative CWR species are: • Vicia sp. (7 species, 114 accessions); • Lathyrus sp. (5 species, 45 accessions); • Sinapidendron sp. (7 species, 34); • Trifolium sp. (9 species, 34); • Hordeum murinum (22); • Beta patula (19); • Medicago sp. (4 species, 18); • Patellifolia procumbens (15).

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Processing Beta patula accessions This process for Beta patula takes in average 88 days and includes: i) Accession starting documentation (attribution of accession number (ISOP)); ii) Sample observation and sanitation (temperature pre-treatment, and separation of

damaged or contaminated seeds); iii) Sample cleaning (separation immature seeds, debris and other foreign material, etc); iv) Sample weighting (up to 3 times) and issuing record samples; v) Sample controlled dehydration in atmosphere moisture controlled conditions); vi) Sample measurement of seed moisture; vii) Germination tests (optinal); viii) Seeds counts and samples selling under vacum conditions; ix) Storage.

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Beta patula germination tests The CWR species can be stored in germplasm collections when produce orthodox seeds. Nevertheless the germination and conservation of theses accessions can face several difficulties, because: i) Germination can be affected by dormancy or other seed protection mechanisms; ii) Storage conditions (namely the moisture conditions) can vary according to species and

be unknown; iii) Germination protocols and conditions can be unknown. The Beta patula accessions has been submitted to 3 different germination tests: a) Soil seed bank samples, in field conditions – 0,8 to 1% viability; b) Soil seed bank samples, ex situ conditions – 10 to 15% viability; c) Species accessions included in the collection, in laboratory conditions – 60% viability Germination protocols used observe ISTA standards and has used between paper or sand techniques.

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Analyzing the species population genetic diversity and its spatial display to delimit the genetic reserve boundaries This study is underway and is based in the use of polymorphic SSRs. 32 SSRs has been identified 14 polymorphic SSRs has been selected 7 SSRs have identified linkage with traits (phenology, behaviour, tolerances) 4 SSRs has already be applied to a sampling of Beta patula populations composed by 150 specimens samples.

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Managment plan of Beta patula reserve The plan will be setup at end and and predicts periodical population monitoring and sampling each 5 years.

PhDs

• Miguel Ângelo Carvalho

(Coordenador)

• Jan Slaski

• Mahnaz Khadem

• Susana Fontinha

• Graça Costa

PhD Students

• José Filipe Ganança (AVRG)

• Teresa Maria dos Santos

• Carla Gouveia (ANT)

• Nuno Nunes (UBQ)

Masters

• Cristina Oliveira

• Marta Rodrigues

Technicians

• Humberto Nóbrega (C + SDI)

• Emanuel Silva (CBM)

• Gregório Freitas (SEAC)

• Sónia Ferraz (ABRG)

Master Students

• Abel Rodrigues

• Magda Santos

34 http://www.uma.pt/isoplexis