I. CONTACT INFORMATION
Organization: INIA
Address: Andes 1365, p 12. Montevideo
Telephone: Fax: 00598-2-9020550
Web page: www.inia.org.uy
http://www.planttreaty.org
Benefit-sharing Fund of the International Treaty
Call for Proposals 2008
PROJECT PROPOSAL FORM
Contact Person
Mr
Name_Francisco L. Last name Vilaró
Position: Director Vegetable Crops National Program
Address: Ruta 48, klm 10, Las Piedras-Canelones
P.O. Box 11100
Country: Uruguay
Telephone: 00598-2-3677641
Fax: 00598-2-3677609
E-mail: fvilaro@inia.org.uy
II. INFORMATION ON THE ORGANIZATION
A. BACKGROUND Maximum 500 words
Outline any previous work done by the organization and its links to this application.
Please provide some information about the rationale behind the project proposal.
Bacterial wilt caused by Ralstonia solanacearum produces many losses in warm potato
climate and more recently it has spread to most temperate regions (CIP 2007). It is
considered the second most important disease for potatoes worldwide, limiting crop
adoption in various tropical and subtropical regions. It causes severe crop losses,
affecting soil for various years and compromising harvested potatoes for seed use
(Hayward, 1991). During 2001, it was found in Uruguay main potato production areas
affecting 39 % of sampled farms (DIEA-MGAP 2003). The only solution has been prevention
measures and there are no resistant varieties available today. Disease resistance
incorporated from various related species was unstable to varying disease and or
environmental conditions (French et al, 1998).
Uruguay is the center of Solanum commersonii, diversity (Correl, 1962). This species
belongs to the wild species pool related with cultivated potatoes (S. tuberosum)
germplasm. This tuberiferous diploid species has been rated as possessing various
valuable biotic and abiotic resistances (Hanneman and Bamberg, 1986). Among these,
high level of resistance to Ralstonia solanacerum, and other pathogens (Erwinia,
Verticillium, Alternaria) affecting the crop (Laferriere et al, 1999). Additionally, it has
been described as a possible source for improving adaptation to cold and drought
environmental conditions (Chen et al, 1999). This species however, possesses some
http://www.planttreaty.org
barriers to its utilization, such as ploidy level, post cigotic incompatibility barriers
explained by the EBN factor (Hanneman, 1994) and its high glicoalcaloid content.
Besides, its level of expression and the genetic basis of resistance to this pathogen is not
known.
INIA, along with National University Faculties (Agronomy and Chemistry) have
undertaken preliminary research on this local solanaceous species. These activities
included genetic and morphological studies along with glicoalcaloid content (Vazquez et al,
1997). Some variability through this species was found, including supposed interspecific
natural hybrids with S. chacoense, poorly represented in our country (Siri et al, 2009,
2005 a and b, Villanueva et al, 2004). True seed from open pollinated collections and
controlled crosses is being kept for short and long term conservation. Additionally,
several clonally propagated introductions are maintained in vitro (Galván et al, 2007).
Exploratory research allowed the collection and characterization of R. solanacerum
isolates, resistance screening methodology and identification of resistant genotypes
(Galván et al, 2007, Villanueva et al, 2004, Dalla Rizza et al 2008) with low glicoalcaloid
content. Also, a segregating population was developed for further genetic and molecular
studies.
Preliminary efforts were initiated for introgression of valuable factors from this species
into the cultivated pool. Controlled crosses with bridge species (S. phureja (2n=2x;
2EBN) succeeded to circumvent cross incompatibility factors associated with this species
(2n=2x; 1EBN), previous to crossing with S. tuberosum (2n=4x; 4EBN). Success rate of
this two crossing phases has been quite low, depending on occurrence frequency of
unreduced gametes (natural polyploidization). (Dalla Rizza et al 2008, Gonzalez et al
2008, Carputo et al 1997). A reasonable number of interspecific hybrids should be obtained to
allow for agronomic traits selection along with high level of resistance to bacterial disease. For
these, polyploidization efficiency (sexual and asexual) should be considered.
Additionally, the development of functional markers could facilitate disease resistance
genes introgression into the cultivated potatoes, along with other adaptation valuable
factors. For this approach, alelic sequence polymorphisms for trait analyzed are needed
(Andersen et al., 2003; Gebhardt et al., 2007). There are various references relative to gene
expression studies based on DNAc analysis for developing useful trait associated
markers. (López et. al., 2006, Restrepo et. al., 2005). A POCI microarray is available to conduct
this research (Kloosterman et al., 2008). The development of SNPs chips would allow the
massive genotyping of resistant individuals.
B. ORGANIZATION
i) Please select from the following options to show the nature of the organization
Government X
A farmer, farming community or farmers' association
Non Governmental Organization
Gene bank or ex situ collection of PGRFA
Science, research & academic sector
Private Sector
A regional or international organization
ii) Maximum 500 words Describe the organization’s mandate, organisational structure,
funding, and capacity to administer small scale projects (organizational and financial
documents may be provided as attachments)
INIA is the main agricultural research institution in Uruguay (www.inia.org.uy). Its mandate is to
promote technological innovation for increasing country competitiveness through
environmentally and socially sustainable production systems. Funding is provided through
agricultural produce taxes, matched by government funds. Several research projects from national
and international sources are executed.
Its organizational structure covers main agricultural products and production systems. A
Directory Board represented by government and grower organization delegates addresses its
research policy, along with several administrative areas. Five experimental stations distributed
along the country, execute research projects belonging to each of 11 National Programs, in
collaboration with 5 Technical Units.
iii) Indicate if a Board or a specific steering committee has been appointed to oversee the
development of this project. No
III. PROJECT DESCRIPTION
A. PROJECT TITLE
Broadening of potato (Solanum tuberosum) genetic basis through introgression of local
wild species, Solanum commersonii .
B. DURATION OF THE PROJECT 24 meses
Start of the project (month/year) June 2009
End of the project (month/year) June 2011
C. SUMMARY OF THE PROJECT Maximum 500 words
Please provide a brief summery of the project identifying the objectives and activities.
The overall objective is to increase genetic variability of potato germplasm. This would
contribute to broadening crop adaptation, through incorporation of disease resistance to R.
solanacearum and other valuable traits from a local wild solanaceous species (Solanum
commersonii).
It is proposed to complete the collection and characterization of Solanum commersonii, including
unreduced gamete production and glicoalcaloid content. In addition the resistance genetic basis to
Bacterial wilt caused by Ralstonia solanacearum will be studied. A segregating S. commersonii
population was developed previously for this purpose, to develop genetic and molecular studies.
Through controlled inoculation with appropriate Ralstonia solanacearum isolates, its disease
reaction will be characterized, on foliage and tubers of S.commersonii and interspecific hybrids
individuals. RT-PCR will be used to quantify more precisely the resistance levels in this
germplasm
Incompatibility crossing barriers will be circumvented through two successive phases of sexual
and or asexual polyploidization and the inclusion of a bridge species (Solanum phureja).
Besides, it will be attempted to introgress this resistance into cultivated potato (Solanum
tuberosum) germplasm assisted by molecular markers to be developed. For the first time in this
species the use of functional markers, based in diferentially expressed genes is proposed. This
approach would be developed on identified resistant individuals, through controlled inoculation
with the pathogen.
Candidate or functional markers are genes suspected to play a functional role in desired
phenotype, such as quantitative pathogen resistance. Allelic variations of these genes could be
responsible for natural variation observed on a given trait. Gene polymorphisms localized within
or physically near could be associated with trait variation.
Finally, agronomic performance of interspecific hybrids will be evaluated for selecting highly
adapted germplasm for further breeding. It is expected that additional environment adaptation
factors from this species would be transferred for promoting more sustainable potato crops. This
should have valuable implications facilitating its development, contributing to increase
availability of this nutritious food worldwide.
D. PROJECT OBJECTIVE(S), ACTIVITIES AND OUTPUTS
Complete the following table:
-
Objective 1: Complete collection and characterization of S.commersonii at national
level
Activities

Project outputs
S. commersonii collected and
characterized for unreduced gamete
production and glicoalcaloid
content
Milestones
and
indicators
True seeds
and in-vitro
clones
described
and
maintained
Due date
12th month

Complete and
maintain
national
S.
commersonii
collection (true
seed and invitro clones)
Characterize
this collection,
including
unreduced
gamete
production and
glicoalcaloid
content
-
Objective 2: Improve R. solanacearum resistance characterization in foliage and tubers
of S. commersonii and interspecific hybrids.
Activities

Project outputs
Characterize
Accessions characterized
resistance level
of
S.
commersonii
and interspecific
hybrids
to
Milestones
and
indicators
Technical
Report and
publication
Due date
12th month


Bacterial wilt.
Quantify by RTPCR presence
of bacterial cells
for
assessing
resistance
levels.
Determine
genetic base of
resistance
to
Bacterial wilt in
S. commersonii.
Highly resistant genotypes
selected
Genetic basis of resistance
determined
Objective 3: Developing functional markers to facilitate disease resistance genes introgression
from S. commersonii species.
Activities
Project outputs
Milestones and
indicators
Plant
inoculation,
plant
sampling
Gene expression determination time.
Micropropagated 1-5th month
plants, disease
inoculation,
RNA
purification
Candidate genes involved determined.
Bioinformatic
data analysis,
primer design
and report.
POCI arrays
analysis
Response
Response analysis functional markers
genes
through RT-PCR in reference
conformation population.
Resecuenciación
de genes de
respuesta
Due date
5-10th month
10-18th
month
Functional
markers
development
SNP search and identification. Chip
design (functional markers <100SNP) Funtional
markers chip
Functional
markers
validated
Functional markers utilization in
germplasm individuals (mapping
populatio, interspecific crossings)
Phenotypic association studies.
Technical
Report and
publication
16-24th
month
Gene introgression from S.
commersonii through functional
markers (chips)
Technical
Report
16-24th
month
Assisted
marker
introgression
Objective 4: Incorporate resistance and general adaptation factors from S.commersonii inpotato
improved germplasm.
Activities
Project outputs
Milestones
and
Due date
indicators
Incorporate
resistance and
general
adaptation
factors from
S.commersonii
in potato
improved
germplasm.
Breaking incompatibility barriers for Interspecific
incorporating
wild
germplasm
into crosses
cultivated one.
obtained
12th month
Potato
Broadening of potato genetic basis through
development of improved population with population
available for
adaptation and resistance factors desired.
24th month
developing
improved
varieties
E. MONITORING Maximum 500 words
Identify who will be the person responsible for the reporting of the project as well as for
monitoring the project progress and its impact assessment according to concrete
milestones and indicators.
Francisco Vilaro will be responsible for monitoring and reporting project progress. He is
Director of INIAs Vegetable Crops National Program. Because of this position, he is
responsible of five research projects through institutional funds, coordinating several
researchers in various disciplines. He has collaborated in several externally funded
projects, national and international, mainly related to potato crop breeding. At present, he
is Principal Investigator of a CGIAR-GCP Project for developing potato cultivars in SE
Africa, involving CIP and various other Program countries.
F. TEAM COMPOSITION AND CAPACITY Maximum 500 words
Give information regarding the team composition. Please specify briefly their discipline
and background and identify if the project foresees the use of available local expertise.
FranciscoVilaró, PhD, plant breeder, cv included
Matías Gonzalez, prebreeding, MSc Student,
Marcos Dalla Rizza, PhD, molecular biologist, cv included
Collaborators
INIA
Diego Maeso, MSc, plant virology
Alicia Castillo, PhD student, tissue culture
University
Guillermo Galván, cv included
María Inés Siri, PhD, biochemistry, cv included
Instituto Pasteur
Pilar Zorrilla, molecular biologist, cv included
INIA´s potato breeding program is being executed for over 25 years. Overall adaptation
to specific cropping system and disease resistance to main pathogens is one of the major
breeding objectives (Vilaró et al 2005, 2006). Major progress has been achieved for virus
resistance and blights (early and late) tolerance. Molecular marker methodology for main
potato virus (PVY) immunity conferring gene (R yadg) is routinely used for progenitor
screening (Dalla Rizza and Vilaró, 2006). Some local cultivars have been developed and
are being adopted.
Through interdisciplinary work approach, our group obtained financial support from
various government and institutional funds for valorization and utilization of this species,
through introgression on the cultivated potato germplasm. These follow:




Control de Ralstonia solanacearum mediante diagnóstico molecular en suelos y
semillas de papa y mejoramiento por resistencia. CSIC Sector Productivo,
Universidad de la República (2003-05).
Creación de un banco caracterizado de germoplasma para mejoramiento de papa e
identificación de factores de resistencia. INIA-LIA BID (2000-2003).
Factores de resistencia en Solanum commersonii para el mejoramiento de papa por
resistencia a Ralstonia solanacearum. CSIC 2005-06.
Estudio multifactorial de la biodiversidad de Solanum commersonii como fuente de
resistencia natural para el mejoramiento de la papa. Conycit-PDT 2005-07.
Recently a collaborative network has been established, funded by CYTED-Spain for
developing strategies for integrated control of Ralstonia solanacearum, causing Bacterial
wilt of potatoes. Our research group is collaborating, along with other countries groups.
Other
collaborative
research
currently
underway
includes:
Papasalud
www.neiker.net/neiker/papasalud/ financed by CYTED. This three year project promotes
potato germplasm characterization and enhancement. Besides, we participate from last
year in a collaborative network (Latinpapa) involving most Latin American countries and
Spain
coordinated
through
CIP
(International
Potato
Center)
(www.cipotato.org/redlatinpapa). This is a valuable platform for adoption of improved
potato germplasm.
G. CONTRIBUTION TOWARDS DEVELOPMENT Maximum 500 words
i) Describe the potential contribution of the project to economic development
In Uruguay is the main horticultural crop comprising around 9.000 has/year. Potato is the first
horticultural crop in Uruguay with an annual consumption over 40 kgs/person. Over U$ 2
million are spent annually for importing potato seed. Risk of Bacterial wilt infection and
poor quality local seed explain this. Improved resistant varieties would largely reduce
imported seed. Besides, during some years considerable crop losses are experienced. For
example, in 2003 these were estimated for over U$ 4 million (DIEA-MGAP).
Additionally it is expected that broad adaptation traits would be contributed from local
wild species, improving crop sustainability and possibly reducing agrochemical use.
ii) Specify with an X if the project contributes to:
* eradicate extreme poverty and hunger
* ensure environmental sustainability
X
iii) Indicate if the project contributes to other Millennium Development Goals1 and how.
Potato is the world´s third most important food crop with an annual production over 300
million tons. Developing countries contribute about 50% of that production (Hijmans,
2001) and will continue to play a key role in feeding billions of people in the coming
years because of its high productivity and nutritional value. Since the early 1960s, it has
outstripped all other food crops in the developing countries in terms of growth in
production area, and this trend is expected to continue. The potato is grown in over 130
countries. Potato is one of the highest value crops in Latin America, whereas Southern
Cone region contributes around 50% to this production (FAO Stat).
The potato yields more nutritious food more quickly on less land and in harsher climates
than any other major crop. The potato is the most important root and tuber crop in the
world. The crop is fundamental in the diets of populations in countries in South America,
Africa, Central Asia and Asia. It is a major carbohydrate in the diet of hundreds of
millions of people in the developing countries. Besides, potatoes contribute to health by
providing valuable nutrients: proteins, vitamins and minerals to the diet. Potatoes are rich
in protein, calcium and vitamin C and have an especially good amino acid balance
(Woolfe 1987).
H. RELEVANCE TO NATIONAL OR REGIONAL PLANS AND
PROGRAMMES Maximum 500 words
Explain the relevance of the project to the country’s or region’s priorities in its plans and
programs for Plant Genetic Resources for Food and Agriculture.
Most regional potato Programs conduct breeding projects to develop improved varieties.
Developed germplasm with broad genetic basis is needed for increasing potato
sustainability production through new adaptative traits.
I. CONTRIBUTION TO CONSERVATION AND SUSTAINABLE
DEVELOPMENT Maximum 500 words
Please identify how the project contributes to the conservation and sustainable use, either
of a specific crop or generally to plant genetic resources for food and agriculture
Worldwide, economic losses from pests and diseases of about €3 billion/year are
estimated for potato (Potato Genome Sequencing Consortium, 2005). The main biotic
constraints are late blight, viruses (PVY and PLRV) and insects. In Brazil, insects alone
(e.g. Diabrotica and potato tuber moth) cause losses of 33% and farmers may spray up to
30 times to control pests and diseases (Franca, 1999). Viruses are especially critical to
small farmers who do not have sufficient resources to renovate their seed frequently.
Potato is also one of the most sensitive crops to water stress (Van Loon,.1981), which
may lower yields by up to 50% (Lahlou et al 2003, Bezzerra et al 1998) especially where
low-input agricultural systems predominate. Potato varieties are not all affected to the
same extent by water shortages due to differences in morphological structure,
1
Find the Millennium Development Goals at: http://www.fao.org/mdg/
physiological behavior and rooting systems (Van Loon,.1981, Lahlou et al 2003, Levy
1986).
Despite the efforts made to date, less than 1% of the variability for disease and pests
resistance present in Solanum species have been used in the present varieties (Potato
Genome Sequencing Consortium, 2005). It is a major challenge to develop potato
cultivars comprising the large array of traits needed in present-day varieties, while
maintaining sufficient variability to meet future scenarios including climate change.
Meeting this challenge would make the crop a more sustainable one for resource-poor
farmers in our region, and in others to which diverse, improved material from our
programs could be adapted.
Over the last 15 years, several such genes have been located on potato linkage maps
using DNA markers, either as major genes or as quantitative trait loci (e.g. Gebhardt and
Valkonen 2001, Gebhardt et al 2006, Feingold et al 2005) which facilitate the
combination and tracing of resistant factors previously introgressed into cultivated
potatoes and also help increase their frequency (e.g. select for cuadruplex) with simple
methods (TaqMan assay, De Jong et al 2003 a, b).
This crop is quite susceptible to various abiotic and biotic pressures, partially attributed to the
narrow genetic basis being used. Drought and temperature extremes comprise major production
crop constraints (Potato Genome Sequencing Consortium, 2005, Franca, 1999). Adaptative
traits of newer national varieties would enable more sustainable production in developing
countries to meet pressing needs for food security and improved incomes with reduced
risk to the environment and human health
Bacterial wilt caused by Ralstonia solanacearum could produce major losses in potato crops,
both in warm and temperate climates. It is considered the second most important crop disease.
Developing bacterial wilt resistant potato cultivars could improve this crop sustainability, mainly
in warm climate regions, reducing direct and indirect losses. Additionally agrochemical use
reduction is expected through more adapted germplasm developed tolerant to various other biotic
and abiotic stresses.
J. PROJECT BUDGET
Benefit-sharing Fund budget:
i) Indicate the funds requested in USD from the Benefit-sharing Fund for each year of the
duration of the project, providing details in the budget notes below. Overhead costs are
excluded.
YEAR 1
1. Staff
2. Supplies
3. Services
4. Local travel
5. Equipment
6. External
Consultant
Total
YEAR 2
TOTAL
8
5
4
3
3
4
8
7
2
1
16
12
6
4
3
4
27
18
45
Budget notes: Please provide details on each of the budget items (i.e. the list of
equipment and supplies, number and type of staff required, purpose and number of trips,
etc)
Staff, 1 research assistant U$ 650/month
Supplies, molecular biology reactants
Local travel, U$ 500/trip for germplasm collections
Equipment, greenhouse irrigation and benches
Services, Ralstonia solanacearum screening, glicoalcaloid determination
Other contributions: Indicate the funds provided by the organization in USD for each
year of the duration of the project, providing details in the budget notes below
YEAR 1
1. Personnel
2. Equipment
3. supplies
4. services
5. communications
Total
14
14
2
3
1.5
YEAR 2
14
14
2
3
1.5
TOTAL
28
14
4
6
3
55
Budget notes: Provide detail on each budget item.
Technicians salaries
Supplies potting, lab reactives
Services, administration, ghouse heating
ii) Indicate if you have applied for co-funding from other sources or submitted this
proposal to other funding sources? No
K. BENEFICIARIES
i) Please select who is (are) the immediate beneficiary (ies) of the project
Government
Farmer, Farming Community, Farmer’s association X
Non Governmental Organization
Gene bank or ex situ collection of PGRFA X
Science, research & academic sector X
Private sector
A regional or international organization
ii) Maximum 500 words. Indicate how the results of the project will directly or indirectly
reach the beneficiaries
Improved germplasm obtained through this project would allow the development of
better adapted potato varieties.
L. COLLABORATION
i) Describe and provide contact information of any collaboration promoted through the
project and explain how it contributes to the effectiveness and efficiency of the project.
Please list collaborators in the table below.
Collaborating Institution
Contact Person
Position
Address
Telephone / Fax
E-mail
Udelar Facultad de Agronomía
Guilermo Galván
Profesor Adjunto de Mejoramiento
Genético Hortalizas (Grado 3), 40 horas,
Departamento de Producción Vegetal de la
Facultad de Agronomía, Universidad de la
República.
Avda. Gral. Garzón 780. CC 12900.
Montevideo. Uruguay
00598-2-359-7191
ggalvan@adinet.com.uy
Collaborating Institution
Contact Person
Position
Address
Udelar Facultad de Química
María I. Siri
Telephone / Fax
E-mail
00598-2-9241881
msiri@fq.edu.uy
Collaborating Institution
Contact Person
Position
Instituto Pasteur, Montevideo
Pilar Zorrilla
Technical Assistant Molecular Biology
Platform, Pasteur Institute
Mataojo 2020, Montevideo. Uruguay
598-2-5220910
zorrilla@pasteur.edu.uy
Address
Telephone / Fax
E-mail
General Flores 2124. CC1157. Montevideo.
Uruguay.
ii) Maximum 500 words. Describe the roles and responsibilities, both technical and
managerial of the institutions and key staff involved in the project.
Francisco Vilaró PhD, project responsable, breeding activities.
Matías Gonzalez M Sc student, screening germplasm for Bacterial wilt disease, breeding
activities.
Alicia Castillo, PhD student , in-vitro propagation and germplasm conservation
Marcos Dalla Rizza PhD, molecular biology activities
Guillermo Galvan PhD student, collection and characterization of wild germplasm
María Inés Siri, PhD, Collection of Ralstonia solanacearum isolates and inoculum
production for screening.
Pilar Zorrilla, M Sc student, microarray methodology.
M. GEOGRAPHIC EXTENSION Maximum 500 words
Please explain how widely applicable (geographic extension) would the results of the
project be and identify the specific countries that would benefit from the project.
Southern Cone South America and most temperate subtropical potato producing
countries worldwide. Many developing countries possess this type of climate and suffer
from Bacterial wilt crop losses. Improved germplasm from this project would be
adaptable to most of those countries.
N. SUSTAINABILITY Maximum 500 words
Please indicate how the project would ensure sustainable activities and beneficial
changes during and after its duration.
Improved germplasm obtained would allow the development of more sustainable
potato varieties.
O. ADDITIONAL INFORMATION Maximum 500 words
Provide any additional information that you may think will be useful in assessing this
Project Proposal.
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ANNEX
IV. DISBURSEMENT
Please provide the following bank account information where payments must be made.
This information must be provided in a separate page and become an Appendix of the
Project Proposal.
CODIGO SWIFT: BSCHUYMM
Account number: CAJA DE AHORRO US$ Nro. 1516540
Beneficiary: INIA LAS BRUJAS
Bank: BANCO SANTANDER
Branch: AGUADA
Address: Av. San Martín 2108
City and country: MONTEVIDEO, URUGUAY