Download GS13048_2015_Report_Potato2015

Adapting Agriculture to Climate Change: Collecting, Protecting, and Preparing
Crop Wild Relatives (CWR Project)
2015 Technical and Financial Progress Report
Project title: Sustainable use of potato crop wild relatives (CWR) and
development of a pre-breeding core collection with key climate
change-related traits
Trust grant no: GS13048
Project reference no:
Project starting date: 1 December
2013
Report due date: 31 January 2016
Project end date: 31 December 2016
Period covered by this report: 1 December 2013 - 31 December 2015
Implementing Institution:
Principal investigator:
Position:
Address:
International Potato Center
Merideth Bonierbale
Senior Potato Breeder
Av. La Molina 1895, Lima 12, Peru
Telephone:
Email:
511-3496017
[email protected]
This report is approved for submission to the Crop Trust by:
_______________________
Signature
_______________________
Name
_______________________
Position
_______________________
Date
A. Technical Report
1. Narrative summary of progress
Describe progress made in implementing the project from commencement to
present. Please include brief descriptions of the approach taken and outcomes
achieved and how other project partners were engaged. (Progress against
workplan should be noted in the Table 1. below).
More systematic use of potato crop wild relatives (CWR) was supported by the
compilation of historical information on wild species and accessions with
resistance or tolerance to biotic or abiotic stresses and the development of a
database of 2n pollen producing 2x cultivated landraces and hybrids with
valuable attributes for use as bridge material for introgression programs. The
resulting databases will be useful for potato breeding aimed at broadening the
genetic base of potato with novel variation conferring adaptation to climate
change. Novel 4x stocks combining disease resistance from 2x CWR, and
adaptation to abiotic constraints from elite 4x breeding lines, have been
selected for agronomical and quality attributes and adaptation to warm climate
and long photoperiod. The most promising pre-bred genotypes will be cleaned
and introduced in vitro for ready incorporation of novel variation into advanced
breeding populations. New CWR-derived true seed (TS) hybrids have been
generated by intercrossing or backcrossing of 4x stocks for subsequent
screening against local pathogen variants and clonal selection for agronomical,
quality, and adaptation traits. For this purpose, a protocol for generation of tuber
families from TS has been documented and uploaded to the project’s website.
A Peruvian student has been trained during 2014 on sample preparation and
identification of meiotic phases, assessment of male fertility and 2n-gamete
production, and the diagnosis of interspecific reproductive barriers.
Report 2015
The consolidation of a potato pre-breeding core collection comprising true seed
(TS) progenies and pre-breeding lines has been accomplished by the
development of 4x TS hybrid stocks from more than 50 crosses between 4x
hybrids derived from 2x sources of late blight resistance from CWR Solanum
Series Piurana, Tuberosa, and Megistracroloba with 4x advanced breeding
lines with virus resistance, earliness and heat tolerance.
Nineteen selected late blight resistant 4x pre-breeding lines, most of which
contributed to the generation of the aforementioned TS stock, were selected for
their good tuber appearance and yield and are currently under in-vitro
introduction for cleaning.
Seven 4x pre-breeding lines developed from bacterial wilt resistance sources
of S. commersonii by INIA Uruguay are in process of introduction to the CIP
Genebank for inclusion in the MLS.
TS progenies from BC3 and BC4 crosses of F1 hybrids developed from sources
of bacterial wilt (BW) resistance from S. commersonii and ancient CIP breeding
lines were dispatched from INIA–Uruguay to EMBRAPA and CIP-Ethiopia to
evaluate stability of resistance against Races 1 and 3 of Ralstonia
solanacearum in endemic areas of the pathogen.
A protocol for evaluation of BW resistance in greenhouse and field conditions
was contributed by our partner from EMBRAPA where it is currently being
applied in screening of TS progenies provided by INIA-Uruguay.
As use of wild species in breeding may result in germplasm with undesirable
high glycoalkaloid contents which can be toxic and impart bitter flavour, hybrid
selections are assessed for total glycoalkaloid concentrations (TGA). We found
less than 9 mg/100g TGA in the 19 selected CWR derived late blight resistant
4x pre-breeding lines introduced in-vitro, among which those involving S.
cajamarquense showed the lowest contents (0.4-5.0 ppm). Concentrations
above 20 ppm are considered unsafe for human consumption.
Twenty four self-compatible (SC) hybrids produced from introgressing the Slocus inhibitor (Sli) gene from S. chacoense to native diploids of Groups Phureja
and Stenotomum with high Fe, Zinc and vitamin C contents were selected for
their tuber appearance, high fertility and ability to produce 2n gametes. These
SC 2x hybrids represent a valuable resource for capturing traits from 2x selfincompatible wild species and developing RILs for genetic mapping and
molecular characterization of novel genes controlling climate change
adaptation traits.
2n-gamete production is a valuable trait of potato genotypes that allows the
transmission of allelic diversity and useful genes from wild species to cultivated
gene pools through sexual polyploidization crossing schemes (4x-2x and 2x4x)). As a first step toward molecular characterization and search of candidate
genes for 2n pollen production, QTL mapping for this trait was performed in the
DMDD segregating population derived from the doubled monoploid DM 1-3
used to develop the Potato Genome Sequence. Consistent QTL across two
and three environments were identified on chromosome 3 of the female parent
DMD and on chromosome 2 of the male parent D, respectively associated with
an increase of 1% of 2n pollen production.
Participatory selection trials have been initiated to evaluate 16 selected late
blight resistant 4x hybrids derived from resistance sources from S.
cajamarquense in two farmer communities in the highlands of Peru by the NGO
Yanapai. Two trials have been set up in the locations of Colpar (Huancayo,3500
MASL) and Pomavilca (Huancavelica, 3870 MASL) using the Mother-Baby trial
design and three local cultivars as control. Traits for which farmers’ opinion will
be relevant are plant vigor, habit and appearance at flowering time, and tuber
appearance, taste and texture at harvest. By the time of this report, the crop is
in flowering stage and the first evaluation underway (February 2016).
The first draft of the 2nd edition of CIP’s Handbook of Techniques and
Procedures in Cytogenetics has been developed in Spanish language and will
be a valuable tool for students, pre-breeders and breeders interested in using
CWR for the introgression of novel genes and broadening the genetic base of
potato populations.
2. Deviations from the project workplan
Note any activities that have had to differ from the original workplan explaining
the consequence of deviations and what was done to alleviate them. Note any
changes to project approach, management, collaborators and risks. Include
any technical issues that have arisen and any constraints to implementation of
activities. Implications to expenditure against the budget should be documented
in Section B of this report.
Report 2014
Modifications to planned procedures for disease resistance screening (bacterial
wilt [BW]) were made during the project inception meeting in Brazil (Nov. 2014,
CIP, INIA, EMBRAPA). This was done because the current protocol developed
by Brazilian partners takes nearly two years, considering that screening is first
performed for race 3 and then asymptomatic plants are allowed to tuberize to
grow plants for assessment of resistance to race 1 under field conditions. We
agreed to maintain a clean copy of each genotype, take stem cuttings from
plants selected as resistant to race 3, and then perform the screening for
resistance to race 1 under greenhouse conditions. Taking into account timing
of quarantine procedures, tuber family development, production of additional
BC3 true seed, and greenhouse screening, we estimate that activity “2.1
Greenhouse screening for BW resistance of BC3 4x progenies derived from S.
commersonii and S. chacoense,” initially planned for this year, will be
completed by June 2016.
Report 2015
Activity 4.2 Semi- controlled greenhouse trials to test temperature effect on 2ngamete production could not be accomplished due to unexpected flaws in the
phytotron growth chamber (Conviron CMP5000 series controllers) where the
experiment was installed. We are looking for a cubicle with semi-controlled
conditions to run the experiment again but new tuber seed will only be available
by July 2016 and the trial will start on September 2016. It is likely that results
won’t be available for the final report.
Participatory variety selection trials have been initiated to evaluate 16 selected
late blight resistance 4x hybrids derived from resistance of S. cajamarquense
in two farmer communities, under the collaborative agreement of CIP and
Yanapai. Two locations were selected for field trials and tuber seed provided
from multiplication plots in Huancani.
Once established and by flowering time several plants in each location were
found to be affected with symptoms of infection with Pectobacterium spp. The
presence of the bacteria was confirmed by CIP bacteriology laboratory in plants
samples taken from the field at Quilcas. The origin of infection seemed to be
the place in the highlands where tuber seed was propagated. Symptomatic
plants will be discarded and hopefully both trials will progress successfully to
harvest. However, it is likely that tubers may be infected and the tuber quality
evaluation that include cooking and organoleptic assessment could not be
performed. We will initiate propagation of tuber seeds of this material in
greenhouse during 2016 to establish a new trial in farmers’ communities by
November 2016 which is the start of growing season in the highlands. We are
hopeful that activity can be completed successfully by April 2017.
CIP’s SSA Potato Breeder based in Ethiopia departed from CIP in mid 2015,
and the position is vacant until a replacement can be found. Likewise, his
research assistant in Ethiopia, where evaluation of families for bacterial wilt
resistance is planned, and CIP-Peru’s prebreeding assistant have both
accepted opportunities for graduate studies in projects led by or associate with
CIP. TS is already planted in Ethiopia and a new assistant has been hired there.
An assistant will be hired in Peru (CIP-HQ) to pick up the project activities
assigned to the person who left.
Despite institutional efforts on both sides, it has still not been possible to
transfer funds from CIP to EMBRAPA for its role in the project.
3. Capacity building
Please comment on progress of any capacity building under the project
Report 2014
An undergraduate student from the Biology Faculty of Ricardo Palma
University (Peru) joined the project as a practitioner in April 2014. She is
acquiring skills in hybridization and the characterization of fertility and
reproductive barriers, including assessment of pollen viability, pollen tube
growth, 2n-pollen, and the preparation of samples to study meiosis. Her
practical period will culminate in March 2015 with a report to the university
and CIP.
Report 2015
The project’s first undergraduate student, Milagros Teruya Kamiyama, from
the Biology Faculty of Ricardo Palma University (Lima-Peru), completed
her one year of practice in Cytogenetics techniques in March 2015 and a
report was presented to CIP and University She gained skills in crossing,
evaluation of 2n-pollen, preparing samples to study meiosis, pollen tube
staining to determine levels of cross compatibility. A new undergraduate
student, Eunice Pena, from the Faculty of Agriculture of Universidad
(Nacional del Centro Huancayo-Peru), joined the project as a trainee on
October 16th 2015. She has already acquired skills on evaluating pollen
viability, 2n-pollen and chloroplast counting for ploidy determination.
Eunice will be involved in the evaluation of water stress tolerance in plants
from putatively resistant wild species that will take place during 2016 under
greenhouse conditions. Her practice will culminate on October 16th, 2016
with a report to the university and CIP.
Our partners from NGO Yanapai have considered their participation in this
project as an opportunity for strengthening capacities. Yanapai has
included a student from the Faculty of Agricultural Science of the Public
University of Huancavelica to be responsible for the coordination of
evaluations with farmers of the field trial at Pomavilca in Huancavelica
along with the group’s regular staff.
4. Information products
List any data and information products arising from this work and describe
where they can be found. We also welcome receiving any photographs of
your work that you may be willing to share.
Database of project material, available in project database, option News
Feed:
https://research.cip.cgiar.org/confluence/display/PotatoCWRprebreeding/
Home
Contents:
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Table 1. CWR derived selected 4x hybrid parents for incorporation of novel
genes for late
blight (LB) resistance in 4x breeding populations (Replaced in 2015)
Table 2. Wild potato species putatively tolerant to abiotic/biotic stresses
Table 3. Wild potato accessions with resistance to LB
Table 4. Wild potato accessions with resistance to BW
Table 5. Putative drought-tolerant accessions from 11 CWR species under
propagation from TS
Table 6. 2n-gamete producing landraces and hybrids
Report 2015
Table 1 Late blight resistant HER clones (Excel File)
Table 7. Amount of seeds and berries in HER X 4x breeding lines &
HERxHER crosses (Excel file)
Table 8. QTL mapping of 2n pollen production in DMDD population (Excel
file)
Table 9. Selected self- compatible BSLI hybrids (Excel file)
Table 10.Total number of CWR genotypes harvested for drought tolerance
evaluation trials CWR_2015” (Word document)
Table 11. CWR_genotypes_under_drought tolerance evaluation trial.
(Word document)
Tuber family generation from 4x BC3 progenies derived from S.
commersonii in Ethiopia (Report from CIP Regional office in Ethiopia, pdf
file)
M_Teruya_R. Palma University Student-Practice Report (Word document)
Handbook Guía de Procedimientos en Citogenética (A Spanish version of
“Manual and Guides in Cytogenetics Procedures”, Word document)
Figures
Figure 1. Potato seedlings transplanted to plastic cups, held in plastic
crates ready for greenhouse inoculation with Ralstonia solanacearum
R3Bv2
Figure 2. CWR derived late blight 4x resistant potato hybrids growing in
fields of farmers’community in Quilcas (Participatory selection trial)
Figure 3. Dr Carlos Lópes from EMBRAPA (Brazil) showing a row of
bacterial wilt susceptible potato clones close to resistant ones in a field
testing trial
5. Publications
List the citations of any scientific papers arising from this work submitted
during the reporting period and planned for the next year
A poster entitled “Broadening the genetic base of resistance to potato late
blight with novel sources from distant wild relatives” was presented at the
11th Solanaceae Meeting held in Bahia, Brazil, on 2–6 November. The
poster has been uploaded to the project’s website at
https://research.cip.cgiar.org/confluence/display/PotatoCWRprebreeding/
Home, option News Feed. This poster describes the generation of novel 4x
stocks with resistance to late blight (Phytophthora infestans) derived from
wild potato species from 2x Piurana Series that have been evaluated for
disease resistance (2013–14) tuber quality traits in 2014 and will be
intercrossed in 2015 to generate TS families for distribution among project
partners and other collaborators.
Report 2015
Planned for 2016:
 A second edition of the Electronic Handbook of Techniques and
Procedures in Cytogenetics will be issued by CIP CPAD service of
CIP (Spanish version)
 A protocol for selection of potato clones for resistance to bacterial wilt
by artificial infection and natural field infection contributed by our
partner Carlos López from EMBRAPA will be translated from
Portuguese to English, edited and issued by CIP CPAD service of
CIP.
6.
Lessons learned
Summarise any lessons arising from the project activities and how these
lessons could/will be used to improve project performance in the future.
The writing of progress reports should begin well before the due date to
allow for timely submission. Partners’ reports will be requested pro-actively
one month before due date and in-house (CIP) collaborators will prepare
regular interim reports to raise awareness of progress and any delays in
expected results.
7.
Complementary/linked activities
Describe any relevant activities underway or planned that link or are
complementary to the CWR Project in general. Give details of the
collaborating institutions.
Development of a Project Website. The aim of this website is to provide
background information on the project and its ongoing activities to the
scientific community. Currently available among project partners, it will be
in the public domain by the end of 2015. The site will provide databases on
genetic material of potato CWR documented in internal institutional reports
and scientific publications from CIP and project partners, successful bridge
species accessions prebreeding material generated previously and under
this project documented for reproductive features, resistance and tolerance,
quality, and nutritional and user preference traits. Protocols for handling
prebreeding material developed as TS, for assessing selected traits for
adaptation to climate change, and a manual describing methodologies and
techniques employed to effectively use potato CWR for trait introgression
to the cultivated gene pool will be accessible to potato breeders and
scientist
through this website.
A multi-institutional research proposal, “Linking genetic resources,
genomes and phenotypes of Solanaceous crops,” was presented to the EU
Horizon 2020 call under the leadership of ENEA (Italian National Agency
for New Technologies, Energy and Sustainable Development) with CIP’s
participation.
Report 2015
The multi-institutional research proposal “Linking genetic resources,
genomes, and phenotypes of Solanaceous crop” presented to the EU
Horizon 2020 was funded and CIP will have an active role in the
prebreeding objective contributing novel pre-bred stocks representing
recent transfer of traits across gene pools and respective phenotypic data
for genotyping along with respective donor accessions
CIP has received funds from USAID to support its breeding and prebreeding activities under a Project entitled: “Advancing Achievements in
Breeding for Early, Resilient and Nutritious Potato and Sweetpotato”. This
project includes among objectives strategies and tools to enhance
selection and trait transfer in and among gene pools. Here, structured
diploid pre-breeding populations will be developed for genetic and genomic
studies. The funding is not new, but represents a modification in modality
of contribution from W1/W2 to W3 funds targeted to breeding.
8.
Progress against Specific Outputs (to be reported in the table
overleaf)
a) For each Specific Output in the project workplan overleaf, describe
progress to date in column E. For those activities/tasks that were due within
the current reporting period, please indicate if these were met and when by
completing column D.
b) If any activities/tasks that were due within the current reporting period were
not met according to the proposed plan, state the reasons why they were
not completed and propose corrective actions in column E and indicate the
revised due date in column F.
Table 1. Project Workplan
Reporting Period: 01/12/13 – 31/12/15
A
B
C
Output
Activities/Tasks
Due Date
(Please do
not modify)
1. Genetic
base of
breeding
population
s
broadened
with
selected
CWR
donor
materials
(Please do not
(Please do
modify)
not modify)
1.1 Generation of
December
BC3 true seed from
2014
BC2 selected S.
commersonii and S.
chacoense derived 4x
hybrids with
resistance to bacterial
wilt (BW), heat
tolerance (HT) and
long photoperiod (LD)
adaptation.
1.2 Generation of TS
from intercrossing of
selected F1 Piurana
derived S. tuberosum
hybrids with late
blight resistance (LB)
and heat tolerance
(HT)
December
2015
D
Actual
Completi
on Date
E
Commentary on Progress to Date
Decemb
er 2014
Eleven second backcross (BC2) 4x hybrids derived from
wild species S. commersonii and some ancient complex
CWR derived hybrid lines bred for BW resistance at CIP
during the 80s and 90s were backcrossed to BW-resistant
4x breeding lines and commercial varieties to develop 30
BC3 TS progenies improved for tuber quality and traits for
adaptation to climate change. In addition, 24 F1, 19 BC1,
and 1 BC2 progenies derived from BW-resistant S.
commersonii and S. chacoense accessions produced by
INIA Uruguay in 2011 are under in-vitro germination for
conserving healthy copies before micro propagated plants
are produced and acclimated for BW screening. Advance
of this germplasm to BC2 will be conditioned upon BW
resistance screening results.
Selected 4x hybrids obtained from crossing LB-resistance
sources from CWR Series Piurana, Tuberosa, and
Megistracroloba to 4x advanced breeding lines with
earliness, potato virus Y (PVY) extreme resistance and
heat tolerance were characterized for LB resistance,
reproductive, and agronomical attributes during 2014 and
compiled in an Excel database (Table 1 at project website,
option News Feed). These hybrids have been designated
as "HER" for “Hybrids derived from Embryo Rescue of
CWR x 2x native cultivars”.
Report 2015 Intercrossing of HER were unsuccessful,
yielding a range of 1 to no more than 15 TS per family.
Therefore, 4x TS families for pre-breeding core collection
stock and distribution to collaborators have been
Decemb
er 2015
F
Revised Due
Date
(if necessary)
A
Output
B
Activities/Tasks
C
Due Date
(Please do
not modify)
(Please do not
modify)
(Please do
not modify)
1.3 Tuber family
generation and family
testing of 4x TS
generated (TS with
BW resistance, HT
and LD; TS with LB
and HT) by CIP
Regional Program in
Ethiopia
December
2016
D
Actual
Completi
on Date
E
Commentary on Progress to Date
F
Revised Due
Date
(if necessary)
generated from crossings of HER back to advanced 4x
breeding lines from CIP’s breeding program. A range from
50 to 2460 TS per family were generated from 82 crosses,
out of which 37 had more than 300 TS/family. By the time
of reporting, berries from 350 new crosses were in
progress of being harvested, as well as additional berries
from 49 crosses, most of which corresponded to crosses
that produced less than 50 seeds previously. Table of TS
and Berry amounts per cross is available in an excel file
named “Table 7. Seed and Berry amounts of HER x 4x
breeding lines & HER x HER” in News Feed of the Project
Website .
https://research.cip.cgiar.org/confluence/display/PotatoCW
Rprebreeding/Newsfeed
A batch of 15 true seed (TS) 4x BC3 and BC4 families
developed by INIA Uruguay’s partners from bacterial wilt
resistance wild accessions from S. commersonii and ancient
CIP breeding lines were dispatched to regional program in
Ethiopia for tuber family generation and field testing for
bacterial wilt resistance assessment and adaptation to local
conditions.` Plants from these families are growing at
greenhouse in order to generate tuber families for field
resistance assessment to bacterial wilt (BW) and clonal
selection. Harvesting of a 1427 successful grown plants from
2750 TS sown are planned by the end of April 2016. A
detailed and illustrated report from our collaborators in
Ethiopia is available in a pdf file in New Feed browse of the
project web page.
A
Output
B
Activities/Tasks
C
Due Date
(Please do
not modify)
2.
Evaluate
and seek
to
compleme
nt new
prebreeding
pools for
key
pathogen
resistance
s, traits
relative to
climate
change,
end-user
and
market
preference
s
(Please do not
modify)
2.1 Greenhouse
screening for BW
resistance of BC3 4x
progenies derived
from S. commersonii
and S. chacoense.
(Please do
not modify)
December
2014
D
Actual
Completi
on Date
E
Commentary on Progress to Date
Report 2014
BC3 TS families developed from selected BC2 4x hybrids
derived from S. commersonii and ancient CIP breeding
lines with BW resistance were field planted for clonal
selection under warm and long photoperiod conditions of
the spring season in Las Brujas (Uruguay), resulting in the
selection of two promising advanced BC3 clones. These
clones have been introduced in vitro. Following the
selection and in-vitro introduction of other BC3 clones,
selected BC3 clones will be assessed for resistance to BW
race 3. Remnant TS from BC3 will be dispatched to
EMBRAPA (Brazil) for screening against BW races 1 and
3 under greenhouse conditions and to CIP’s regional
program in Ethiopia for tuber family generation and field
testing for adaptation to local conditions. If insufficient
seed is available for distribution to both places, priority will
be given to Brazil, and the cross will be repeated to
generate additional seed for Ethiopia and for seed stock.
Taking into account timing of quarantine procedures, tuber
family development, production of additional BC3 TS, and
greenhouse screening, we estimate completion of this
activity by June 2016.
Report 2015
Two batches of TS families from BC3 and BC4 crosses
derived from S. commersonii and ancient CIP breeding
lines with BW resistance were received by Carlos Lopes at
EMBRAPA from Francisco Vilaró (INIA, Uruguay). The first
batch of 13 families, was sown in trays with sterile soil and
the seedlings transferred to 1.5 L pots with sterile
substrate in a screenhouse. Two to five the tubers from
F
Revised Due
Date
(if necessary)
June 2016
A
Output
B
Activities/Tasks
C
Due Date
(Please do
not modify)
(Please do not
modify)
(Please do
not modify)
2.2 Field assessment
of selected BW
resistant BC3
December
2015
D
Actual
Completi
on Date
E
Commentary on Progress to Date
each clone were harvested 100 days after transplant.
Approximately 100 clones were obtained from each of the
13 families. These clones are in the cold room and will be
planted in a field naturally infested with R1B1 of Ralstonia
solanacearum in 2016. The second batch of seeds
consisted of 12 families. They were sown in trays, like
before, and were transplanted to 250 mL plastic glasses,
which were inoculated with a suspension of R3B2 of R.
solanacearum. Plants were maintained in a greenhouse
heated at night in order to guarantee plant infection, thus
lessening the chances of escapes. The surviving
seedlings from approximately 100 seedlings inoculated in
each family were transplanted on Dec 10, 2015 to 1.5 L
pots filled with sterile soil mixture. They will be kept in a
screenhouse to produce tubers, which will be challenged
with the R1B1 of the pathogen in March/April 2016. A table
with the percentage of infection surviving seedlings from
each family is available in EMBRAPA report 2015Screening of 4x BC3 progenies for resistance to BW
uploaded in News Feed
https://research.cip.cgiar.org/confluence/display/PotatoCW
Rprebreeding/Newsfeed of the project Website
Percentage of infection surviving seedlings among families
ranged from 4 to 19%. The greatest value corresponding
to a BC4 family derived from combining BW resistance
from S. commersonii and ancient resistant bred lines from
CIP.
Twenty nine BC3 clones representing 9 families were
selected out of 82 during Fall crop season at INIA Las
Brujas (Uruguay). Seven BC3 clones representing 4
F
Revised Due
Date
(if necessary)
A
Output
B
Activities/Tasks
C
Due Date
(Please do
not modify)
(Please do not
modify)
genotypes for
agronomical
attributes, heat
tolerance and long
photoperiod
adaptation
2.3 Field assessment
of selected LB
resistant 4x Piurana
derived hybrid lines
for agronomical
attributes and heat
tolerance.
(Please do
not modify)
2.4 Total
glycoalkaloid content
analysis of tubers
from selected CWR
D
Actual
Completi
on Date
E
Commentary on Progress to Date
families were further selected under heat and long day
length conditions of spring (October) in the same location.
December
2015
December
2016
A total of 163 selected late blight resistant 4x hybrids
derived from resistance sources from CWR Series
Piurana, Tuberosa, and Megistracroloba (HER clones, see
Activity 1.2) underwent field selection for agronomical
attributes in La Molina (Lima, Peru). Dry matter and pollen
fertility were also determined. Nineteen hybrids showed
moderate to high yields (≥0.7 K/plt) and dry matter content
(>20%). Most of these selections come from late blight
resistance sources from S. cajamarquense
(SeriesTuberosa) (67%), whereas the remaining ones
come from S. chiquidenum (21%) and S. paucissectum
(11%), both from Piurana Series. The selected group
showed to be highly fertile (pollen viability >80%), only 9
hybrids showed moderate fertility (>50 - <80%). A table
with information from the 163 late blight resistant HER
clones is available in an excel file “HER clones_2015”
(Table 1) in News Feed of the Project Website
https://research.cip.cgiar.org/confluence/display/PotatoCW
Rprebreeding/Newsfeed
The use of some wild species may potentially cause a useful
increase of the dry matter content of tubers and an
undesirable increase of glycoalkaloids (GA). Although the
upper limit allowed for a new cultivar release is 20 mg/100g
fresh weight, bitterness can be tasted in tubers with
F
Revised Due
Date
(if necessary)
A
Output
(Please do
not modify)
B
Activities/Tasks
(Please do not
modify)
derived 4x selected
clones
2.5 On farm trials to
assess farmer’s
preference of
selected hybrid
clones
C
Due Date
(Please do
not modify)
December
2016
D
Actual
Completi
on Date
E
Commentary on Progress to Date
F
Revised Due
Date
(if necessary)
glycoalkaloid levels higher than 14mg/100g (Sinden et al.
1976).We analysed GA in 40 out of the 163 selected late
blight resistance HER clones (see Activity 2.3). GA content
ranged from 2.3 to 56.0 mg/100g in samples of unpeeled
tubers and from 0.4 to 36.5 mg/100g in samples of peeled
tubers. The 19 HER clones with good yields, dry matter
content and fertility reported in Activity 2.3 reached GA
contents less than 14 and 9 mg/100g in unpeeled and peeled
tuber samples, respectively. The lowest contents were found
in selections from S. cajamarquense (range= 0.4-5.0) making
them useful for future breeding efforts. The selected group of
19 HER clones have been introduced in-vitro to initiate their
cleaning for future availability and distribution.
Participatory selection trials have been initiated to evaluate
16 selected HER clones in two farmer communities in the
highlands of Peru, i.e., Quilcas (Region Junin; between
3400-3800 MASL) and Acobamba (Region Huancavelica,
3867) under the collaborative agreement of CIP and Yanapai
as part the accomplishment of CWR project activities. Two
trials have been set up respectively, in the locations of
Colpar (Huancayo,3500 MASL) and Pomavilca
(Huancavelica, 3870 MASL) using a Mother-Baby design
and three local cultivars as control. The two trials were
planted in November 2015, and will be harvested by the end
of April 2016. Traits for which farmers’ opinion will be
relevant are plant vigor, habit and appearance at flowering
time, and tuber appearance, taste and texture at harvest. By
the time of the report, the crop was by its flowering time and
several plants were found visually affected with symptoms of
infection with Pectobacterium spp. The presence of the
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3. A CWR
derived
prebreeding
core
collection
for traits
related to
climate
change
adaptation
, i.e., heat
and
drought
tolerance
and key
pathogen
resistance
s, i.e., to
Late blight
(LB) and
3.1 Compile
information of CWR
accessions
documented with
resistance to BW, LB
and/or tolerance to
heat and drought
December
2014
D
Actual
Completi
on Date
Decemb
er 2014
E
Commentary on Progress to Date
bacteria were confirmed by CIP bacteriology laboratory in
plants samples taken from the field at Quilcas. Symptomatic
plants will be discarded and hopefully plants will reach a
satisfactory harvesting. Yanapai has considered this
collaboration as an opportunity for strengthening capacities
by involving a student Faculty of Agricultural Science of the
Public University of Huancavelica to be responsible of the
field conduction and coordination of evaluations with farmers
of the experimental trial at Pomavilca in Huancavelica.
Historical information on CWR species and specific
accessions with BW or LB resistance, or putative tolerance
to drought ,was compiled from internal institutional reports,
thesis research, scientific publications, or consultants’
contribution from CIP, gathered into Excel databases and
uploaded to the project ‘s website (Tables 2–4, option
News Feed). Project partners from INIA-Uruguay and
EMBRAPA contributed information on 4x advanced hybrid
lines developed by their corresponding breeding programs
using BW resistance from S. commersonii and S.
chacoense, or from introductions of CIP breeding
germplasm during the 90s. Ten 4x advanced hybrid lines
are anticipated to be delivered to CIP during this and
coming project years to shape the prebreeding core
collection. Databases will contribute valuable information
for potato breeding aimed at broadening the genetic base
with novel variation for traits associated with adaptation to
climate change. Some CWR species have already been
used to generate 2x pre-breeding material with resistance
to LB and BW, and have been in turn deployed in elite 4x
genetic background under this project. In contrast, no
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Bacterial
wilt (BW),
of potato
developed
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3.2 Vegetative
propagation of
accessions from invitro plants
December
2014
D
Actual
Completi
on Date
E
Commentary on Progress to Date
previous evaluation has been done to identify droughttolerant accessions among putative sources suggested by
former CIP scientists and compiled here. Therefore,
drought tolerance will be tested and confirmed in a sample
of accessions of these species before trait transfer is
attempted into cultivated backgrounds aimed at
contributing to the enrichment of pre-breeding core
collection.
Of 38 species suggested by CIP’s former scientists as
likely sources of drought tolerance, 21 were selected
based on endosperm balance number or effective ploidy to
ensure success in crosses with cultivated species. TS from
each of 3 accessions of 19 species was available and
provided by CIP’s Genebank, out of which TS from 1 to 3
accessions of only 11 species germinated successfully
(Table 5 in project’s website, option News Feed) and are
currently growing under greenhouse conditions in CIP’s
highland station in Huancayo (3,249 masl) for tuber seed
production.
Report 2015
Twenty three putative drought tolerant accessions
identified from 11 wild potato species during 2014 were
sown as TS in greenhouse conditions in 2015 to get
tubers for drought tolerance trials . Tubers were harvested
from 719 genotypes. (A table of total number of harvested
genotypes is available in a word document named “Table
10.Total number of CWR genotypes harvested for drought
tolerance evaluation trials CWR_2015” in News Feed of
the Project webpage
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https://research.cip.cgiar.org/confluence/display/PotatoCW
Rprebreeding/Newsfeed)
3.3 Greenhouse trials
for resistance or
tolerance
assessments, as
appropriate
December
2015
Report 2015
Of 719 genotypes harvested from aforementioned
vegetative propagation activity (Activity 3.2), a sample of
471 that yielded 10 or more tubers were selected. After 8
weeks, tubers from only 10% of genotypes broke dormancy
and therefore a chemical product had to be applied to
accelerate sprouting. By October 2015, 400 genotypes
sprouted evenly. Ten tubers per genotype were planted in
trays until plants reached 10 cm tall. A first experimental
trial for water stress response evaluation has been set using
280 genotypes and 5 potato cultivars with different levels of
response to water stress as controls, Desirée (CIP 800048,
90 day maturity) regarded as tolerant compared with other
European cultivars; Sarnav (CIP 3970077.16, 120 day
maturity) regarded as a high yielding and drought tolerant
cultivar; Tomasa Condemayta (CIP 720072, 160 daymaturity) regarded as a drought sensitive cultivar; Qonpis
(CIP 700921, 180 day maturity) as moderately drought
tolerant cultivar from Group Andigenum, and one wild
potato (Solanum commersonii, CIP 761102) reported
previously as drought tolerant .
The greenhouse pot experimental trial for drought tolerance
evaluation was set up at CIP Experimental Station in
Huancayo (3259 MASL). A table with the list of testing
genotypes is available in a word file entitled “Table 11.
CWR_genotypes_under_drought tolerance evaluation trial”
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word file in News Feed of the Project webpage
https://research.cip.cgiar.org/confluence/display/PotatoCW
Rprebreeding/Newsfeed). Four plants from each of the 280
wild potato genotypes and controls were planted in pots
containing 5 kg of wet sand. Two plants of each genotype
were used for the water stress treatment and the remaining
two for the well water treatment. Pots were fertilized with
250 ml of Peter 7-7-7 (3 gr/l) one week after planting and
with 250 ml of “Grow More 20-20-20” (3g/l) weekly. Damage
caused by the fungus Rhizoctonia solani were observed in
several genotypes of Solanum bukasovii, S. lignicuaule and
S. infudibuliforme. So far, since the installation of the
experiment (December 2015) temperatures at greenhouse
in Huancayo reached 30 °C to more centigrades and
therefore, watering has to be performed twice a week.
Trait evaluations will take place after hilling, at flowering and
at harvest. So far, evaluations of number of surviving plants,
plant vigor, plant height, stem number, number of lateral
branches, leaf number, chlorophyll content (SPAD), and
plant health were already recorded by the time of reporting.
Plant wilting will be evaluated 20 and 40 days after drought
stress. Tuber appearance, uniformity and physiological
disorders, harvest index, as well as root length and structure
will be evaluated at harvest.
Water-stress treatment will start after flowering initiation.
Water stress will be performed by watering plants with only
half of the volume used for watering plants not subjected to
water stress (Intermittent Drought Irrigation). In addition, the
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3.4 Assessment of
pollen fertility, 2n
gamete production
and self-compatibility
of selected CWR
accessions
3.5 Crossing Block
for trait introgression
to 2n- gamete
producing landraces
or directly to 4x
breeding lines for 2ngemete producing
CWR
4. Abiotic 4.1 Compile
factors
phenotypic data for
stimulating disease resistances,
2n-gamete stress tolerance,
production post-harvest and
identified
nutritional traits of 2nand
gamete producing
applied in CWR and landraces
prebreeding
D
Actual
Completi
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parenchyma type, xylem diameter and pilosity or hairiness
will be determined.
Pollen has already been collected from flowers of the 280
testing genotypes from Activity 3.3. Pollen fertility and 2n
gamete production will be assessed during February 2016.
Self-pollinations are underway gradually as genotypes
flower.
December
2015
December
2016
December
2014
Decemb
er 2014
2n-gametes have been used by breeders to access wild
species germplasm and transfer genetic diversity and
valuable traits from diploid hybrids to the tetraploid level.
Genotypes that produce 2.5% or more 2n-pollen can be
regarded as good 2n gamete producers and hence
suitable for breeding by sexual polyploidization. We have
compiled information on 2n-pollen producer 2x cultivated
landraces from Phureja and Stenotomum groups and
derivative hybrids from crosses with wild species from
Piurana, Tuberosa, Conicibaccata, and Megistracroloba
series. This list (Table 6 in project’s website, option News
Feed) also provides information on tuber characteristics,
resistance, and quality attributes for use in selecting
desired 2n gamete-producing genotypes for pre-breeding
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4.2 Semi- controlled
greenhouse trials to
test temperature
effect on 2n-gamete
production
December
2015
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(if necessary)
purposes.
December 2n-gametes have proven essential in the genetic
2016
improvement of cultivated genotypes, allowing the
transmission of allelic diversity and of useful genes from wild
species to cultivated gene pools through sexual
polyploidization crossing schemes. This study aims at
investigate temperature effects and photoperiod on male
unreduced gamete production in 2x landraces of Groups
Phureja and Stenotomum. A sample of 8 landraces with a
wide range of 2n-gamete production i.e., low= 0-.0.4%,
moderate= 0.8-2.0% and high ≥2.5% were selected. A
factorial design with three factors, cultivars (8 levels),
temperatures (four levels: cold (10 °C), cool (18°C/10°C
day/night), warm (21°C/15°C) and hot (25°C/18°C)), and
photoperiod (two levels: 12 h and 16 h) was planned in a
completely randomized design (CRD) with 40 experimental
units (plants) and 5 observations (plants) per treatment.
Plants were grown in pots under 20 h photoperiod for 30
days before moving to a phytotron growth chamber
(Conviron CMP5000 series controllers) under light intensity
from 11’700 to 18’300 lux. Because temperatures in the unit
unexpectedly fluctuated 5 degrees above or below the
planned temperatures the experiment could not be continued
due to flaws in the appliance. Only the cold treatment at 12 h
photoperiod was performed. However, results cannot be
considered as most plants did not flower. Cultivars CIP703762 and CIP- 704040 known to produce a moderate
frequency of 2n-gametes i.e, 1.4 and 1.5% respectively, at
greenhouse conditions in Huancayo, produced 2n gametes
at a frequency slightly lower than usual, i.e, 0.5 and 1.2%,
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4.3 Mapping of QTL
for 2n gamete
production in the
DMDD map derived
from the doubled
monoploid DM 1-3
used to develop the
Potato Genome
Sequence
December
2015
D
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(if necessary)
respectively. On the other hand, pollen mother cells were
observed in stained anthers of flower buds collected from
high frequency 2n-gamete producing cvs. CIP-703506 and
CIP-703559 (>5%) that aborted their flowers in this trial
conditions. The observation of dyads and triads in these
cultivars suggests potentially 2n- gamete formation. These
abnormal cells have been associated with 2n- gamete
formation previously (Camadro, et al. 2008). The experiment
will be repeated next year in a cubicle with temperature
control.
2n pollen production was evaluated in the DMDD
segregating population generated from the doubled
monoploid DM 1-3 used to develop the Potato Genome
Sequence and the heterozygous diploid clonal accession
(CIP 703825, referred to as D) belonging to the
Solanum tuberosum diploid Group Goniocalyx. DMDD is a
backcross population that used the D clonal accession as
recurrent parent. Evaluations were performed in plants grown
in the greenhouse and field conditions of the highlands in
Huancayo and Paucartambo and lowlands subtropics in Lima
(Peru) from 2009 to 2012. 2n gamete production was
determined as the percentage of 2n pollen in a sample of
200 stained pollen (viable pollen) QTL mapping was
performed on the hybrid female DM/D and D maps
independently using the software JoimMap® 4. Frequency
distribution of DMDD for 2n gamete was skewed to the right
due to a high frequency of genotypes with lack or very low
percentage of unreduced gametes. Therefore QTL analysis
used the Kruskall Wallis statistics. QTL were identified on
chromosomes 1,2,5, 6 and 7 in the male parent and on
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5. A prebreeding
strategy to
facilitate
the
characteri
5.1 Introgression of
Sli gene originating
from S. chacoense
into 2n-gamete
producing diploid
landraces and
December
2014
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(if necessary)
chromosomes 3, 5, 6, 7 and 9 in the female parent DMD.
The proportion of trait phenotypic variance explained (R2) by
this QTL varied from 6% for QTL detected in Acomayo 2013
to 14% for QTL detected in la Molina 2011 in the female
parent DMD. QTL on chr 2 of the male parent DI was the
most consistent across environments followed by that on
chromosome 3 of the female parent DMD. Several of the
QTL detected by means of Kruskall Wallis were also
detected by Interval Mapping when data was transformed to
arc sine. Most of QTL identified were still significant when
data was averaged over environments. An excel file named
“Table 8 QTL mapping of 2n pollen production in DMDD”
showing a table of significant QTL and a chart of the
frequency distribution of genotypes for this trait can be find in
News Feed of the Project website
https://research.cip.cgiar.org/confluence/display/PotatoCWR
prebreeding/Newsfeed QTL mapping is the first step
toward fine mapping and identification of markers for
assisted selection. The Potato Genome Sequence is a
valuable tool to search for candidate genes and new
approaches, such as genome editing, can provide the final
proof of the importance for these candidate genes by
elucidating their function and their actual association with the
phenotype
Dec. 2015 Report 2014
To develop self-compatible (SC) diploids from landraces,
the S-locus inhibitor (Sli) gene known to alter selfincompatible (SI) to SC plant was introgressed from two
SC accessions of S. chacoense into 17 2x landrace
cultivars from the SI Phureja and Stenotomum groups.
A
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zation and
incorporati
on of
genes
controlling
target
traits, and
exploitatio
n of
transgress
ive
segregatio
n or
heterosis
outlined
B
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selected selfincompatible CWR
derived diploid
hybrids
C
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These landraces were selected for moderate-to-high
content of vitamin C, iron, and/or zinc. The percentage of
2n-pollen in 17 diploid landraces ranged from 2.4 to 3.6%
(mean = 2.9%). The unreduced gametes are important for
transferring novel genes to the 4x cultivated genetic pool
by sexual polyploidization. Hybrid TS was successfully
obtained for 26 out of the 36 crosses performed. To date,
Sli introgression has been confirmed in 5 out of the 26
successful crosses. Three out of the five progenies
segregated more than 89% SC plants, while the remaining
two segregated 53% and 72% SC plants, respectively.
Seedling tubers will be recovered at harvest from SC
hybrids from the five progenies and from SC plants of TS
progenies from the remaining crosses. Sli bearing welladapted hybrids derived from 2x landraces will be used as
sources for capturing traits from 2x wild SI species and
development of novel SC hybrids for developing RILs as a
valuable public resource for the mapping of novel genes
controlling climate change adaptation traits and for
exploiting heterosis and transgressive segregation.
Report 2015
Self-compatible (SC) 2x hybrids obtained from
introgression of the S-locus inhibitor (Sli) gene from S.
chacoense to native diploids were grown till tuberization.
Tubers were obtained from a total of 136 SC hybrids under
greenhouse conditions, out of which 24 were selected
based on their high pollen viability, number of tubers and
ability to form 2n pollen. These selected Sli bearing SC
hybrids have been designated as BSLI. A table of these
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6.
Capacities
for prebreeding
developed
5.2 Tuber generation
from self- compatible
plants of CWR- and
landrace- diploid
derived F1 hybrid
progenies
December
2015
5.3 Outline a nested
mating design and
propose a prebreeding strategy
6.1 Capacity in prebreeding techniques
December
2016
December
2016
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BSLI is available in News Feed (Table 9 Selected selfcompatible BSLI hybrids) BSLI of the Project website
https://research.cip.cgiar.org/confluence/display/PotatoCW
Rprebreeding/Newsfeed
December Two selected self-compatible BSLI hybrids from activity 5.1
2016
were used to develop a nested mating design using sources
of late blight resistance from S. circaeifolium var.
capsicibaccatum, S. commersonii, S. microdontum, S.
sogarandinum and S. tarijense. BSLI hybrids were used as
male parents to pollinate 88 late blight resistance genotypes
from 8 different accessions from the aforementioned
species. By the time of this report, 200 to 1900 seeds/cross
were obtained in crosses with 8 late blight resistant
genotypes from two accessions, one from S. microdontum
and the other from S. tarijense. Crosses will continue during
2016 to generate seeds or otherwise increase the number of
seeds in crosses with other accessions and species.
Generation of tuber families from this crossing block will be
postponed until succeeding on a Nested Mating Design
whose progenies represent as most accessions and species
as possible depending on cross compatibility.
Report 2014
An undergraduate student, Milagros Teruya Kamiyama,
from the
Biology Faculty of Ricardo Palma University (Lima-Peru),
joined the project as a practitioner on 1 April 2014. She is
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and new
plant
genetic
resources
available
in public
domain
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E
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gaining skills in diagnostic techniques of fertility and
prezygotic barriers such as pollen viability and pollen tube
growth. Milagros has also been trained in crossing,
evaluation of 2n-pollen, and in preparing samples to study
meiosis. Her practice will culminate on March 20th 2015,
with a report to the university and CIP.
Report 2015 Milagros completed her one year practice
gaining skills on procedures in Cytogenetics and a report
was presented to CIP. This has been uploaded to the
Project web page as “M_Teruya_R. Palma University
Student-Practice Report”. A new undergraduate student,
Eunice Pena, from the Faculty of Agriculture of
Universidad Nacional del Centro (Huancayo-Peru), joined
the project as a trainee on 16 October 2015. She has
already acquired skills on evaluating pollen viability, 2npollen and chloroplast counting for ploidy determination.
Eunice will be involved in the evaluation of water stress
tolerance in plants from putatively resistant wild species
that will take place during 2016 under greenhouse
conditions. Her practice will culminate on October 16th,
2016 with a report to the university and CIP.
6.2 Develop an
updated protocol for
generation of tuber
families from TS
December
2014
December The project aims to develop a prebreeding core collection
2014
from introgression and incorporation of traits from CWR
novel sources in the form of TS. A training manual that
describes each step of how to handle TS material for
producing healthy tuber families has been developed for
partners, collaborators and scientists on how to test
families for resistance or tolerance to biotic and abiotic
stresses related to climate change. A first version is
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publicly available in CIP’s Global Trial Data Management
System at
https://research.cip.cgiar.org/confluence/display/GDET4RT
/Protocols.
6.3 Develop an
updated protocol for
drought tolerance
assessment
December
2015
6.4 Develop a
protocol for bacterial
wilt resistance,
implementing a test
sensitive method for
detection of R.
solanacearum (e/g/
RT-PCR) in latentlyinfected tubers
6.5 Manual of
crossing procedures
updated with mentor
pollination and
embryo rescue
methodology, flow
cytometry analysis,
procedures for EBN
determination and
December
2015
December
2015
Report 2015
A practical guide for screening potatoes for drought tolerance
will be updated with experiences gained from our ongoing
greenhouse pot experimental trial (Activity 3.3) for assessing
wild potato plants for resistance to water stress. This will
contribute guidelines for preparing material, planting,
growing and caring wild plants to be subjected to water
stress, as well as, relevant measuring traits
April 2016 Our partner from EMBRAPA (Brasil) contributed a protocol
for selection of potato clones for resistance to bacterial wilt
by artificial infection and natural field infection. In addition a
protocol for preparing tubers and stem cutting samples to
determine latent infection by PCR were also provided. The
protocol is in Portuguese and will be translated in English
and edited by April 2016.
April 2016 A second edition of the “Handbook of Techniques and
Procedures in Cytogenetics” has been contributed by CIP
scientists Biol. Ms. Sc. Benny Ordoñez and Dr. Merideth
Bonierbale . The handbook will be a valuable tool for student
and plant breeders. Its content comprises topics on the
reproductive biology of potatoes and cytology techniques and
procedures to determine pollen viability and fertility, ploidy
determination, 2n gamete production, pollen tube growth in
pollinated pistils, embryo rescue and culture. The Handbook
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methods for
determining
pollination behavior
through examination
of pollen-pistil
interactions
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will be issued in Spanish, we expect to have an English
version by end of 2016. The first draft is available in News
Feed of the Project Website under the name of “Manual y
Guía de Procedimientos en la Citogenética”, we recommend
to use the correct citation when referred to a technique used
from this Manual and to respect the rights of authorship
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References
Sinden SL, Deahl KL, Aulenbach BB. 1976. Effect of glycoalkaloids and phenolics on
potato flavor. Food Sci 41(3):520-3.
Camadro, E.L., Saffarano, S.K., Espinillo, J.C., Castro, M., Simon, P.W. (2008).
Cytological mechanisms of 2n pollen formation in the wild potato Solanum okadae and
pollen-pistil relations with the cultivated potato, Solanum tuberosum. Genetic
resources and crop evolution. 55(3):471–477.
B. 2015 Financial Progress Report - International Potato Center (CIP)
Category
(Please do not modify)
1.Personnel
2.Supplies
3.Services
4.Equipment
5.Travel
6. Admin (20%)
7. Consortium fee
Total CIP
Total EMBRAPA
Total INIA
Total Yanapai NGO
Grand Total
Total
Cumulative
Budget
Expenditure
(Please
2013 do not
2015
modify)
134,236
61,662
36,103
13,388
147,007
87,274
0,0
0,0
46,971
24,588
72,884
35,972.83
10,718
5,049
447,899
227,933
43,200
0,0
43,200
20,273
12,301
1,405
546,600
249,611
Budget
2015
Actual
(Please Expenditures
do not
2015
modify)
49,218
28,142.10
13,603
8,213.60
57,300
50,476
0,0
0,0
15,996
6,995.55
27,224
17,326
3,960
2,657
167,301
113,810
14,250
0,0
14,250
20,273.00
6,150
1.445
201,951
135,488
Provide justification for any variance in use of funds relative to original budget line items
greater than 10%:
Notes to the Financial Report
Please provide detail on actual expenditures below.
1. Detail personnel expenditure incurred and the amount of time spent on the project.
2. Provide a list of supplies purchased (items may be grouped where appropriate) and
services received.
3. Provide details, cost and purpose of each trip taken.
4. Provide detail on administrative costs.
5. Provide detail on consortium fee
In kind Contribution (USD) - International Potato Center (CIP)
Category
(Please do not modify)
2. Supplies
3. Services
6. Admin (20%)
TOTAL
Total Budget
(Please do not modify)
6,583
76,467
16,610
99,659
2013 - 2015 Actual
Expenditures
4,782
68,049
19,416
92,247
2015 Financial Progress Report (USD) - Empresa Brasileira de Pesquisa Agropecuária
(EMBRAPA)
Category
Total Budget
Budget
Cumulative
(Please do not modify)
(Please do not
2015
Actual
Expenditure
modify)
(Please Expenditures
2013 do not
2015
2015
modify)
2.Supplies
12,400
4,000
3.Services
30,800
10,250
TOTAL
43,200
14,250
EMBRAPA
2015 Financial Progress Report (USD) - Instituto Nacional de Investigación Agropecuaria
(INIA), Uruguay
Category
Total Budget
Budget
Cumulative
(Please do not modify)
(Please do not
2015
Actual
Expenditure
modify)
(Please Expenditures
2013 do not
2015
2015
modify)
2.Supplies
12,400
11,487
4,000
8,974
3.Services
30,800
8,786
10,250
5,543
TOTAL
43,200
20,273
14,250
14,517
2015 Financial Progress Report (USD) - Yanapai NGO
Category
Total Budget
Cumulative
(Please do not modify)
(Please do not
Expenditure
modify)
2013 2015
2.Supplies
3.Services
TOTAL
7,150
5,151
12,301
735.98
668.90
1,440.88
Budget
2015
Actual
(Please Expenditures
do not
2015
modify)
4,150
735.98
2,000
668.90
6,150
1,440.88
This financial report is
Prepared by:
Certified by:
__________________
Name
Position
_____________________
Name
Position