EUROPEAN COMMISSIONDIRECTORATE-GENERAL FOR AGRICULTURE AND RURAL DEVELOPMENT Directorate H. Sustainability and quality of agriculture and rural developmentH.1. Environment , GMO and genetic resources

Brussels, 12.04.2010 – ARES: 187187C40 001/2/3/2009-202446- need for rev. of TR 2010H.1/od – Save .10.12-D-202446

Subject: Grant 001 AGRI GEN RES 870/2004 "Leafy vegetables germplasm, stimulating use" - Request for a revised third annual technical report

Ref.: Your letter dated 17 March 2010 received on 18 March 2010 by the Commission (Ref: ARES 145501) Our letter dated 17 March 2010 (Ref: ARES: 148179)

Dear M. Dijkhuizen,

We acknowledge the receipt of technical and financial reports of the second year for the Grant 001 AGRI GEN RES 870/2004 "Leafy vegetables germplasm, stimulating use" received on 17 March 2010 by the Commission.

In my letter on 19 March 2010, we have asked for the missing excel table breaking down the manpower by partner, work package, and staff categories. Following the assment by an independent expert, we would like you to deal with the omissions and shortcomings listed below in a revised report.

WP1- task 1: clarify how many data points were added in 2009 in the four data bases. Provide more information on the passport data. Provide a guide on how to use the "project data files". Clarify the link between characterisation data and the database. Clarify the use of the log files.

WP1- task 4: clarify whether problems of quarantine pests and diseases have been encountered, and if so, how those problems were overcome. Provide more information on the safety duplication.

WP1- task 5: clarify how the gap analyses will be carried out. WP2 – task 4: clarify why all regenerated accessions exceed one hundred

percent and explain the difference between task 3 and task 4. Clarify why only around 1000 accessions were chosen for regeneration and the methodology used for the regeneration.

WP2 – task 5: clarify the methodology used for task 5.

WP3 – task 2: provide more details on task 2.

REGISTEREDStichting Dienst Landbouwkundig Onderzoek – DLOAtt. M. DijkhuizenCosterweg, 50NL 6701 BH WageningenTHE NETHERLANDS

Commission européenne, B-1049 Bruxelles / Europese Commissie, B-1049 Brussel - Belgium. Telephone: (32-2) 299 11 11.Office: Loi 130 10/189. Telephone: direct line (32-2) 2961040. Fax: (32-2) 2966255.

E-mail:Olivier.Diana@ec.europa.eu

WP4 – task 3: provide details on the trial carried out by P07. Clarify whether the partners are working for the various WPs according to a collegiate approach across the entire project. Clarify the consequences of the heterogeneity of many accessions for the plant morphology. Clarify the methodology used for organising the public tasting days in organic food stores.

WP5 – task 1: provide the complete minutes of the third WP meeting and of the third annual project meeting as annexes.

WP5 – task 2: provide details concerning the progress of task 2. WP5 – tasks 3 and 4: review the text concerning tasks 3 and 4. WP5 – task 4: clarify the methodology used for task 4. Assessment of methodologies of WPs: provide details of the methodologies of

the WPs and tasks. Progress in the implementation: provide details for milestones 09, 10, 19, 30,

38, 50 and 51. Problem(s) encountered: clarify some of the minor problems encountered. Dissemination of the interim results: clarify the missing references to the

Leafy Veg action 001 and GENRES Community Programme for the web publications. Clarify the coordination of the dissemination of interim results.

In addition to these points, the annex includes also recommendations are for the next reporting period.

Based on the above-mentioned observations and the attached recommendations, we are looking forward to receiving a revised technical report. Please, submit this revised technical report as soon as possible, but not later than 12 May 2010.

In summary, the Commission has two consecutive periods of 45 calendar days for assessing the reports and executing the payment. Please note that Article II.15.3 stipulates that, if additional information is requested, the time limit for the scrutiny shall be extended by the time it takes to obtain this information.

We would appreciate to receive an electronic version of the revised technical report in advance.

Yours sincerely,

Martin SCHEELEHead of Unit

Attachment: extracts of the independent expert's evaluation

O. DianaH.1

D. De BruyneH.1

P. HirschfeldtH.1

A. MorealeH.1

M. ScheeleH.1

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Attachment

Below are listed the observations from the expert's evaluation report and the resulting recommendations.

General comment

"This report uses much the same format, and often much the same words, as previous reports; a simple analysis reveals that 63% of the text of this report is identical to that of the first annual report. It seems that to make this third year’s report, the coordinator has simply taken his the preceding year’s report, changed some wording, added other wording, and updated the figures…

Much fuller details will be required in the revised report and also in the final report for the next reporting period…

The coordinator should receive a stern warning that in the final report his minimalist reporting style must change".

Recommendation: The annual and final reports shall be considered as stand alone documents. The revised technical report shall be significantly improved. The weakest point of the report is the missing descriptions on methodologies used for WPs and tasks. The minutes of the project meetings shall be entirely provided as annexes of the revised report. For the minutes of the meetings, the mention to the link of the coordinator's web site is not sufficient. The use of cut and paste for the content of key tasks (e.g. for WP5-tasks 3 and 4) is not acceptable.It is the second year that a revised report is requested following the observations of two different independent experts. The coordinator shall consider the above-mentioned remarks and the additional recommendations requested below for the revised report.

Concerning the comments of the reviewer:

That 63% of the wording is identical to the previous year is not surprising as one should realize that tasks have already been finalised earlier in the project, so it would be a bit strange to chance the texts of these tasks. Furthermore also for ongoing tasks this reasoning applies to a considerable extent.

Fuller details are provided in the revised report for the various WPs.

A minimalistic reporting style: it is our intention to give as much detail as we think is necessary.

Concerning the recommendations:

The third annual technical report has been updated now; missing descriptions of methodologies, not earlier presented in previous annual technical reports, have been added.

Minutes of the annual and WP coordinator meetings have been added (see annex 4).

The text on WP5 tasks 3 and 4 has been updated.

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We have taken the remarks and additional recommendations into account.

1. WP1 – Documentation

Task 1: Collecting of passport data

"The Report tells us that these four databases now contain passport data of a total of 17,530 accessions. The Report does not tell us how many data points were added this year".

Recommendation: the coordinator shall clarify in the revised report how many data points were added in 2009 in the four data bases.

This info is now included in the report.

"It is a pity that the coordinator does not tell us any more; where did the passport data come from? How were the data verified? Who did the work?".

Recommendation: some explanation was provided in the 2nd technical report. However, the coordinator shall provide more information on the passport data for the revised report.

The collection of passport data already started in the first project year, and is a continuous activity towards the end of the project. Procedures how to collect passport data were outlined in the technical report of 2007. In the present report we included a reference to the methods described earlier. Contributors of the data are included as a separate web page in each of the four databases together with their contact details. We do not see the relevance of copying this info into the annual report. Considering the magnitude of the databases, verification of each provided data point is virtually impossible. As in all central crop databases, it is considered the responsibility of the data provider to submit reliable data. Evidently, the database work was carried out by the respective database managers, i.e. GEVES, IPK and CGN as mentioned in the project description. We did not consider it necessary to repeat the task description in the annual report.

"The“project data files” that are available on the project website appear to contain the results of trials done during the Leafy Veg project to assess winter hardiness, Bremia resistance, etc…It would have been helpful if the report had included a guide to these “project data files”. It would be even better if it was explained how to use these files: how do I find out what is known about the lettuce varieties ‘Dubacek’, ‘Flekica’, and ‘Gelb Roter aus Zagreb’? The data files would also be improved if they included some elements of statistical analysis: what are the confidence limits for a difference between cultivars?".

Recommendation: the coordinator shall consider these remarks for the revised report.

Concerning the project data files we included information where the files can be found. In the beginning of 2010 we included a new functionality in the databases in order to have easier access to the data. Since this was done recently, this was

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not included in the 2009 report. Although this activity was carried out in 2010, we added info about this new functionality in the 2009 report. Because the primary purpose of the databases is to make the basic data available to potential users, statistical treatment of the data goes beyond the main goal of the databases.

Task 3: Uptake of generated data

"The report says that characterization data (from WP2) have been linked to the database. It would have been helpful if the report would have provided a bit more about this because I do not know what “link” means. Probably it means that the characterisation data are in the database and can be retrieved from it by an SQL query. But nowhere is this said. This sort of operational should be obligatory in the final report to the Commission - as it would be in any scientific publication…

In order that all the information produced in the project is easily available, the information in the “downloadable project data files” should be incorporated in the databases and retrievable from there by SQL query. (I do not know if this is already the case or not; the Report contains no explanations.)…

There does not appear to be any analysis of the log files (who visited, when, what were they looking for, etc…). The coordinator should put this in hand now, so that he can tell us in the final report how the leafy veg databases are being used. (Who is searching them? Where do they come from?? Does their search result in a request for material?)"

Recommendation: the coordinator shall clarify the link between characterisation data and the database in the revised and the final reports. He shall clarify the use of the log files in the revised report.

As mentioned in the response to the previous review remark, the link between passport data and data from the characterization, evaluation and utilization workpackages has been established in the databases and a description of this new functionality has now been described in the 2009 report.

Task 4: Arrangement of safety duplicates

"The work on safety duplication of collections appears to be good, but I do not know the details of how it has been carried out. I am thinking of other projects, on other species, where the need to eliminate all quarantine pests and diseases before shipment has effectively blocked exchanges of material. It would have been interesting to know whether Leafy Veg partners have met such problems, and if so, how those problems were overcome. (Were quarantine organisms such as Lettuce mosaic virus and Lettuce infectious yellows virus (EPPO A1 list, EU Annex 1/A1) a hindrance to exchange of material?, etc…)…

Recommendation: the coordinator shall clarify in the revised report whether problems of quarantine pests and diseases have been encountered, and if so, how those problems were overcome.

In the beginning of the project a workplan was developed for the arrangement of safety duplicates (see technical report 2007). Part of this workplan was to make an inventory among the project partners of bottlenecks in the arrangement of

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safety duplicates. No quarantine issues whatsoever were reported by the partners. Main bottlenecks reported were the lack of sufficient seed quantities and/or the workload involved, as already indicated in the 2007 report. We included a brief statement in the 2009 report in order to accommodate the reviewer’s concern.

"It is not possible to check the work done against the contract, for the relevant milestone M09 is so vague as to be useless (it reads “Safety duplication of part of the collections realized”)…

Collections that were unique, and therefore at risk, have now been duplicated (I am not sure whether in part, or in total). Partners have agreed how the final gap analysis will be performed. This is all good. I just regret that more details are not provided".

Recommendation: the coordinator shall provide in the revised report more information on the safety duplication.

We are not sure what additional information about safety duplication is actually requested. In the 2009 report we mentioned which partners introduced a safety duplication arrangement, and we provided an overview table indicating the current status (hosting institutes, storage conditions, potential hosts, what part of each collection has been safety duplicated).

Task 5: Identification of collection gaps

"A quick check (I looked at the Spinach database) reveals that data on latitude and longitude of collecting site are missing for more than 80 percent of the accessions (1666 blank entries, 2013 entries total). Moreover, where the data are available, they are not always to the accuracy (one second of Latitude/Longitude) required by the group’s own protocols. It is therefore not clear to me how the gap analyses will be carried through".

Recommendation: the coordinator shall clarify in the revised report how the gap analyses will be carried out.

Also here we do not understand what additional information is actually requested since the workplan was included in the 2009 report. Our feeling is that the remark of the reviewer is for some reason based on the assumption that latitude and longitude data are absolutely necessary to carry out a gap analysis. Such data mostly apply to collected landraces and crop wild relatives but not to cultivars that usually are the most frequent in crop databases. Moreover, in cases where exact geographical data are missing, it may still be clear from which country, region or place the material originated. The case of spinach mentioned by the reviewer is an example of a crop collection in which wild relatives are severely underrepresented. This observation by itself already contributes to the gap analysis. Evidently, this will be quantified in more detail in the intended publication. As outlined in the presented workplan, structuring of the crop gene pool, identifying main origin areas for cultivated material and main distribution areas for wild crop relatives and subsequent examination how these are represented in the databases will allow a gap analysis.

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2. WP2 – Regeneration and characterisation

Task 4: Regeneration and Characterisation for all crops started

"The coordinator reports that 108% of all accessions involved in the project have been regenerated (91%) and/or characterized (124%) within the three years of the project. He does not comment on why the achievements exceed one hundred percent (perhaps he calculates that two partners regenerating the same accession count as a 200% regeneration)…

I’m not clear how Task 4 differs from Task 3…

Since there are now more than 17000 accessions in the databases, it would have been interesting to learn why “only” a thousand or so were chosen for regeneration. It would also have been interesting to learn how regeneration was done (how were plants isolated? how were diseases controlled?? how was diseases harvested, conditioned and packeted?)".

Recommendation: the coordinator shall clarify in the revised report why all regenerated accessions exceed one hundred percent and explain the difference between task 3 and task 4. He shall also clarify why only around 1000 accessions were chosen for regeneration and the methodology used for the regeneration.

Around 1000 accessions were chosen for regeneration because this is a number which can be handled by the staff during four years. A second limitation factor is the field or greenhouse area because enough space must be available and some of the material must be isolated (see multiplication protocols). Despite these points it was possible to regenerate more material than planned because the staff and the place especially in the Dutch genebank were available. This means more than 100% were fulfilled. Each partner has regenerated their own material, this means accessions were not regenerated twice or double.

In the description of Task 3 was a mistake in the report, this was corrected (see below).

For Task 4 the number of regenerated accessions during 2009 which was done by the involved partners was included in the report (see below). In addition, the multiplication protocols for all crops were described (see below).

Task 3:

In 2009 the partners started with regeneration of the following material pending the growing season:

P0: 16 accessions of spinach were sown for regeneration P1: 20 accessions of lettuce and 6 accessions of chicory were sown in

January, 5 accessions of spinach were sown in march P2: sowing of 17 accessions of chicory for regeneration P3: Corn salad, chicory and 103 accessions of lettuce were sown in

march and 6 accessions of rucola were sown in April P4: no regeneration is required

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P5: sowing of 3 lettuce accessions and 1 spinach accession for regeneration

P6: lettuce accessions for regeneration were sown in February P7: regeneration task completed in 2008 P8: sowing of 17 lettuce accessions for regeneration P9: no regeneration required P10: no material was sown P11: no regeneration is required (As agreed between P3 and P11 P3 will

regenerate 100 chicory instead of P11.)

Task 4:

In 2009 the partners regenerated the following material:

Regeneration 2009

Partner   Lettuce Spinach Chicory Valerianella Rucola

P0 CGN 0 16 - - 0

P1 HRI 20 5 6 - 0

P2 GEVES 0 - 17 0 -

P3 IPK 103 - 39 11 6

P4 Palacky - - - - -

P5 NGB 3 1 - 0 0

P6 INRA 74 - - - -

P7 Arche Noah 0 0 0 0 -

P8 HDRA 17 - - - -

P9 UNIBO - - - - -

P10 KIS 0 - 09 (from

USOFA) -

P11 ProspecieRara - - 0 - -

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Regeneration was done recording to the following protocols:

Multiplication of lettuce ( Lactuca sativa L.)

Isolationo All self-pollinating species, e.g. Lactuca sativa L. are multiplied in field. o Only for wild cross-pollinating species like Lactuca perennis, Lactuca

tatarica, Lactuca siberica and some Lactuca viminea accessions isolation is necessary. Different methods for isolating plants are possible, for example distance isolation, time isolation, caging, bagging and hand pollination.

o Other wild species and accessions are multiplied separately in greenhouses.

Sowingo Usually sowing is performed in spring every year. Seeds are sown into

pots in the greenhouse and later transferred to trays. Plants that lag considerably behind in growth may be left out because their poor performance could mean that these plants eventually do not contribute to seed multiplication of the accession.

o Two month after sowing plants are transferred to field. Cultivation

o If necessary plants are irrigated and treated with fertiliser for optimal growth.

o At commercial maturity accessions are characterized on the basis of descriptor lists (see Task 5).

o Weed control is necessary to limit competition and reduce weed-borne pathogens and insects. Phytosanitary methods are practices like monitoring insect populations, applying biological controls, using clean cultural practices and applying pesticides at appropriate times. These practises are used in both the field and the greenhouse to decrease pesticide use.

Pollinationo Cross-pollinating species like Lactuca perennis, Lactuca tatarica, Lactuca

sibirica and some Lactuca viminea accessions are pollinated by insects (depending on isolation method).

Harvesto Plants are harvested when harvest maturity is achieved, mostly in late

autumn. o All plants of each accession are cut and put into bags. After drying in a

drying chamber, threshing and sieving all accessions are cleaned by hand.

Multiplication of spinach ( Spinacia oleracea L.)

Isolationo Spinach is a cross-pollinating species, consequently isolation is necessary.

Different methods for isolating plants are possible, for example distance isolation, time isolation, caging, bagging and hand pollination.

Sowingo Usually sowing is performed directly onto field in spring in Europe every

year. o

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Cultivationo If necessary plants are irrigated and treated with fertiliser for optimal

growth. o At commercial maturity accessions are characterized on the basis of

descriptor lists (see Task 5). o Weed control is necessary to limit competition and reduce weed-borne

pathogens and insects. Phytosanitary methods are practices like monitoring insect populations, applying biological controls, using clean cultural practices and applying pesticides at appropriate times.

o After commercial maturity spinach accessions need a bird protection. Pollination

o During flowering time plants are pollinated by wind or insects (depending on isolation method).

Harvesto Plants are harvested when harvest maturity is achieved.o All plants of each accession are cut and put into bags. After drying in a

drying chamber, threshing and sieving all accessions are cleaned by hand.

Multiplication of Leaf chicory ( Cichorium intybus L.)

Isolationo Leaf chicory is a cross-pollinating species, consequently isolation is

necessary. Different methods for isolating plants are possible, for example distance isolation, time isolation, caging, bagging and hand pollination.

Sowingo Usually sowing is performed in autumn in Europe every year. Seeds are

sown into pots in the greenhouse. Plants that lag considerably behind in growth may be left out because their poor performance could mean that these plants eventually do not contribute to seed multiplication of the accession.

o All accessions overwinter in pots in the greenhouse. Cultivation

o In next spring plants of each accession are isolated.o If necessary plants are irrigated and treated with fertiliser for optimal

growth. o At commercial maturity accessions are characterized on the basis of

descriptor lists (see Task 5). o Phytosanitary methods are practices like monitoring insect populations,

applying biological controls, using clean cultural practices and applying pesticides at appropriate times.

Pollination o During flowering time plants are pollinated by bees (depending on

isolation method). Harvest

o Plants are harvested when harvest maturity is achieved. o All plants of each accession are cut and put into bags. After drying in a

drying chamber, threshing and sieving all accessions are cleaned by hand.

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Multiplication of Endive ( Cichorium endivia L.)

Isolationo Leaf chicory is a cross-pollinating species, consequently isolation is

necessary. Different methods for isolating plants are possible, for example distance isolation, time isolation, caging, bagging and hand pollination.

Sowingo Usually sowing is performed in spring in Europe every year. Seeds are

sown into pots in the greenhouse and later transferred to trays. Plants that lag considerably behind in growth may be left out because their poor performance could mean that these plants eventually do not contribute to seed multiplication of the accession.

Cultivationo Later in spring plants of each accession are transferred to field. If

necessary plants are irrigated and treated with fertiliser for optimal growth.

o At commercial maturity accessions are characterized on the basis of descriptor lists (see Task 5).

o Phytosanitary methods are practices like monitoring insect populations, applying biological controls, using clean cultural practices and applying pesticides at appropriate times.

Pollination o During flowering time plants are pollinated by insects (depending on

isolation method). Harvest

o Plants are harvested when harvest maturity is achieved. o All plants of each accession are cut and put into bags. After drying in a

drying chamber, threshing and sieving all accessions are cleaned by hand.

Multiplication of Rocket (Eruca spp.)

Isolationo Rucola is a cross-pollinating species, consequently isolation is necessary.

Different methods for isolating plants are possible, for example distance isolation, time isolation, caging, bagging and hand pollination.

Sowingo Usually sowing is performed directly onto field in spring in Europe every

year. Cultivation

o If necessary plants are irrigated and treated with fertiliser for optimal growth.

o At commercial maturity accessions are characterized on the basis of descriptor lists (see Task 5).

o Weed control is necessary to limit competition and reduce weed-borne pathogens and insects. Phytosanitary methods are practices like monitoring insect populations, applying biological controls, using clean cultural practices and applying pesticides at appropriate times.

Pollinationo Flowering time starts two month after sowing and plants are pollinated by

insects (depending on isolation method). Harvest

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o Plants are harvested when harvest maturity is achieved. All plants of each accession are cut and put into bags. After drying in a drying chamber, threshing and sieving all accessions are cleaned by hand.

Multiplication of Corn salad (Valerianella spp.)

Isolation o Corn salad is both a self-pollinating and cross-pollinating species,

consequently isolation is necessary. Different methods for isolating plants are possible, for example distance isolation, time isolation, caging, bagging and hand pollination.

Sowingo Usually sowing is performed directly onto field in early autumn in Europe

every year. Cultivation

o If necessary plants are irrigated and treated with fertiliser for optimal growth.

o At commercial maturity accessions are characterized on the basis of descriptor lists (see Task 5).

o Weed control is necessary to limit competition and reduce weed-borne pathogens and insects. Phytosanitary methods are practices like monitoring insect populations, applying biological controls, using clean cultural practices and applying pesticides at appropriate times.

o Plants overwinter in field with a layer of fir branches. o In the next spring plants start bolting and later flowering.

Pollinationo Corn Salad is a self-pollinator but insects often visit the flowers, therefore

an isolation of accessions is compulsory (depending on isolation method). Harvest

o Plants are harvested when harvest maturity is achieved.All plants of each accession are cut and put into bags. After drying in a drying chamber, threshing and sieving all accessions are cleaned by hand.

In 2009 the partners characterised the following material:

Partner   Lettuce Spinach Chicory Valerianella Rucola P0 CGN 0 - - - 17P1 HRI 11 10 - - -P2 GEVES - - 31 - -P3 IPK 148 - - 10 35P4 Palacky 0 - - - -P5 NGB - 10 - 19 ATRI 0P6 INRA - - - - -P7 Arche Noah 0 12 - - -P8 HDRA 17 - - - -P9 UNIBO - - - - -

P10 KIS 0 - 09 (from

USOFA) -P11 ProspecieRara - - 94 - -

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Task 5: Collecting characterization data of all partners

"After collection, the data were entered into the database. I would have been interested to know how this was done. (Pencil, paper and type by hand? Or electronic data capture and automatic transfer to the database? What sort of sanity checks to detect errors and omissions?) So many questions occur to the reader of the report, and so few answers. I repeat my reminder that such detail will be required in the final report. This work is all worth publishing, so if the partners include all this missing detail in the final report the work will be half done".

Recommendation: the coordinator shall clarify in the revised report the methodology used for task 5. He shall also consider these remarks for the final report.

Methodology for regenerationDuring commercial maturity the partners characterise/describe the material on the basis of the descriptor lists (see above). They go to the field or in the greenhouse and write down their observations in specific lists. After characterisation is finished the documented data are entered in an excel sheet with the agreed format. Each crop has a special format available on the leafy vegetables website. All partners send their duly completed characterisation data to work package leader who is responsible for visual checking partner’s data of accuracy and completeness. If necessary the respective partners will consulted. All checked characterisation data are sent to the coordinator who is responsible for transferring the characterisation data into the database via electronic data capture.

3. WP3 – Evaluation

"With regard to methodologies used in WP3…It turns out that the missing information was already provided by the partners in their Excel files, which are available on the project website. The coordinator has simply failed to copy that information into his report. I can find no reason, nor any excuse, for this failure…

Recommendation: the coordinator shall provide in the revised report more details on methodologies in WP3.

Seven studies have been carried out in 2009 within WP3 on different species and on different characteristics. Therefore, there were seven completely different methodologies used by the different partners. As scheduled and explained in the past annual reports and the Grant Agreement AGRI GEN RES 001, some of these methodologies are the same for studies scheduled on the two or three years program and some other methodologies have been applied in a single year. Nevertheless, as already exposed in past Annual reports, all the used methodologies are included in the respective Excel files (leaflet “Experiment description”) giving, for each study, the aim, the methodology and, of course, the results. By the way, the methodologies are included in the POWERPOINT presentations shown during the Annual meetings (see at http://documents.plant.wur.nl/cgn/pgr/leafyveg/members%20only/cesenameeting.htm; username: leafy and password: popeye ).

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a) For the evaluation of winter hardiness of 6 lettuce accessions (Austrian lettuces), P7 used the following methodology. During the 2008/2009 winter, the accessions were sown in two different locations in Austria (with different climatic conditions); the plants have been put in open field and covered by a fleece as well in a plastic tunnel. Observations and records have been noticed on the ability of these accessions to overpass the Austrian winter (number of survived plants, development of plant disease, harvest maturity period).

b) For the evaluation for Bremia lactucae resistance of 30 accessions (Czech lettuces), P4 used in 2009 the following methodology which is the same than the ones used in 2008. Eleven B.l. races and one isolate have been tested in 2009 (vs three B.l. races in 2008) on a classic protocol: inoculation in growth chamber by each of the races/isolate on two replications of 25 seedlings at fully expanded cotyledon leaves. Two varieties standards have been used as susceptible controls in order to check the test. For each accession, a sporulation intensity has been calculated (see the Excel files at http://documents.plant.wur.nl/cgn/pgr/ildb/download.htm )

c) For the evaluation for Perenospora farinosa resistance of 263 spinach accessions, P0 used in 2009 the same methodology to test such accessions for 2 P.f. races than the one used in 2008 to test the first 4 P.f. races on the same 263 accessions. In glasshouse, 30 seedlings per accession were inoculated two times (at cotyledon stage and when 70-80% of the accessions had one well developed leaf). The protocol of Jim Corell (2003) was used for scoring. In general, 30-48 plants were scored per test. All tests were carried out in duplo. Scores of the duplos were averaged, unless the difference in resistance scoring was more than five classes. (see the Excel files at http://documents.plant.wur.nl/cgn/pgr/spinach/download.htm )

d) For the chemical evaluation in 2009 of 2 new spinach accessions and 16 orache, P5 used the same methodology to test such spinach and Atriplex accessions than the one used in 2008 to test Eruca and spinach accessions. Plants have been grown in glasshouse and 3-5 plants per 10-15 plants have been picked to be tested for dry matter, nitrate content and oxalic acid contents. Dry matter is determined in leafy vegetables gravimetrically after drying at 105°C overnight. For nitrate content, leaves without stalks are chopped, frozen, defrosted and homogenized in two parallel samples analysed; each sample is cooked, Carrez I and II solutions are added, after that the solutions are diluted and filtered and nitrate content is determined by HPLC. For the oxalic acid content, samples are analyzed in duplicates, fresh or frozen; then different chemical products are used in order to get this content by HPLC. Methodology and results are available at:

http://documents.plant.wur.nl/cgn/pgr/spinach/download.htm http://documents.plant.wur.nl/cgn/pgr/minorlv/download.htm

e) For the evaluation of 23 chicory accessions for Alternaria cichorii resistance, P2 used the same methodology than the one used in 2008 on the first 22 accessions. Methodology, accessions and results are available at http://documents.plant.wur.nl/cgn/pgr/chicory/download.htm). About the methodology, plants grown in field after a growing period of the seedlings in greenhouse were inoculated two times by Alternaria cichorii spore suspensione (one just after the plantation, the second three weeks after). A

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susceptible accession has been planted too. Observations has been conducted two weeks after second inoculation and based on a scale of notation (0 to 9).

f) For the evaluation in 2009/2010 of 17 accessions for Sclerotinia sclerotiorum resistance (results finalised in February 2010), the same methodology than the one used in 2008/2009 on 14 accessions. Plants have been sown in field by the P2 partner (FNPE: French Federation of Witloof Producers) during spring period. Roots have been uprooted in November and then forced in specific growth chamber with expression of hot and wet conditions. Their inoculation has been done with an infected piece with Sclerotinia sclerotiorum incolum during the forcing period in this growth chamber. A scale of notation has been used (0 to 5) and a disease index calculated for each accession.

g) For the evaluation of 85 accessions for Thielaviopsis basicola resistance, the whole program has been began in 2009 and finalised in 2010 (two waves of accessions). These accessions of which two standards (one resistant and one susceptible) have been sown in pots and artificial contamination of the soil by this black root rot conducted two days after. A scale of notation has been used: 0 (no symptom) to 4 (decrease of the leaf growth & roots totally necrotic or dead plant). A Necrotic Resistance Index has been calculated. All the results will be shown with the results of the second waves of accessions finalised in 2010.

Task 2: Evaluation of the leafy vegetables material selected

The coordinator should give much more details about the work. (For example he mentions new Bremia lactucae resistance. Readers will want to know if this is a new single gene resistance to a new pathovar? or a new source of durable resistance to all pathovars?)".

Recommendation: the coordinator shall provide in the revised report more details on task 2.

As previously explained, all the detailed information about the studies are included in the Excel files which are useful for scientists and other users of the data in order to do in their conditions such experiences. In order to answer questions of the reviewer: below more information on the experiences conducted in 2009.

a) For the evaluation in 2009 for Bremia lactucae resistance of 30 accessions, twelve other races and isolates have been tested : eleven races (B.l. : 1, 12, 14, 15, 18 , 20 to 25) and one isolate DEG 2, highly virulent originated from Germany but not included in the official set of B.l.. In 2008, three B.l races have been tested (B.l. 5, 16-17).

b) For the evaluation for Perenospora farinosa resistance of 263 spinach accessions, P0 tested in 2009 two races of P.f.s. (i.e. Pfs 1 and Pfs 4). As exposed in the past annual report, P0 tested in 2008 the first two races as scheduled: Pfs 5 and Pfs 7. In 2010, three other races are tested: Pfs 3, Pfs 6 and Pfs 10. Theses seven Pfs races represent the most relevant races in the commercial field production and for which spinach breeders must have to find and develop resistant varieties.

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"In this presentation, species, partners and evaluation variables are all mixed up. I get the impression that each partner is doing their own thing, and that there are few or no interchanges of techniques or material or information across the partnership. I trust that this is not the case, and that everything is coordinated. Perhaps the coordinator could rethink his presentation for his final report; his overall plan would become apparent if the information were presented as a table (rows = species, columns = tests)…

Recommendation: the coordinator shall consider these remarks for the final report.

The whole complete program of the Leafy Veg is included in annex 1 of the Grant Agreement AGRI GEN RES 001 with an overview of the detailed work per partner. For all the work already done, please find below a synthesis per leafy vegetable and per Leafy Veg partner (named P0 to P10). Subjects have been described above.

Some further clarification:

- for lettuce, “104 % (109/105)” means that 104 % of the scheduled has been realised during the three years of the four years program and that 109 accessions on a total of 105 initially scheduled have been tested ;

- for lettuce, for P4, “30 acc. (12/20 B.l. races)” means that P4 has tested all the scheduled 30 accessions until 2009 but only on 12 B.l. races and isolate on a total of 20 Bremia lactucae. races and isolate ;

- for spinach, for P0, “263/250 acc. (4/7 P.f.)” means that P0 has tested 263 accessions (more than the 250 initially scheduled program) by the end of 2009 but only on 4 Perenospora farinosa races ;

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- for chicory, P9 has begun in 2009 sesquiterpene lactone content in 100 accessions but the method to recover and quantify lactones on such chicory has been assessed by the end of 2009.

4. WP4 – Utilisation and marketing

Task 3: Activities on utilisation and marketing

"Partner 7 did a trial with 15 accessions of lettuce, grown on 4 farms...Details of trial size, statistical treatments of results, etc, are not given".

Recommendation: the coordinator shall provide in the revised report details on the trial carried out by P07.

In 2009 P07 organised a field evaluation for the 15 lettuce varieties at the experimental station of Höhrere Lehr und Versuchsanstalt A-2344 Schönbrunn. This was carried out in 3 cultivation periods, summer, spring and autumn. Yield data of spring and summer cultivation could not be taken because of early boltig in spring and flood water at all fields of the experimental station in summer destroying all seedlings of summer cultivation. Experimental design: two replications with 66 plants per accession and plot. Sowing date: 30.07.2007. Planting date: 18.08.2009. Silty soil, pH-value 7,0. Fertilisation: 50 kg Agrobiosol for 448 m².

"For Partner 10...the results are presented in full, as tables, complete …this detail is welcome. It would have been nice if the coordinator had presented results of other work in other WP in similar detail".

Recommendation: the coordinator shall consider this remark for the final report.

This remark will be considered for the final report.

"Partner 11 did one field trial that involved 52 accessions of chicory, many provided by partners 2 and 11. This is good – I would like to see a similar collegiate approach across the whole project. Partners should not be working on their own".

Recommendation: the coordinator shall clarify whether the partners are working according to a collegiate approach across the entire project.

The collegiate question is across the whole project - i.e the gene banks are providing seed for P11 (ProSpecieRara) for WP3 as well as WP4, also P8 (HDRA) had seed of lambs lettuce and rocket accessions from P3 (IPK) for WP3 as well as WP4. Similarly ProSpecieRara (P11) has received seed from genebanks to carry out characterization studies on chicory accessions in WP2. The approach by partners has been collegiate when ever the opportunity has arisen, but for WP 4 it was specifically decided by partners that they primarily wished to mainly evaluate accessions from their own collections as the objective of the WP was to evaluate for local markets.

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"On page 29, the report states that: “Many accessions were heterogeneous for plant morphology”. The coordinator really ought to discuss the implications of this. If accessions are heterogeneous, how are they to be described, recorded, conserved, and regenerated? What does the database say about heterogeneous accessions?".

Recommendation: the coordinator shall clarify in the revised report the consequences of the heterogeneity of many accessions for the plant morphology.

Partners assessing marketability of accessions for this task were also recording accessions for morphology and other traits of interest to growers (e.g. P07- winter hardiness). This information is reported under WP3 for the purposes of this report, however, when uploaded to the relevant database all data on an accession are provided in one file so that users can access all relevant data for an accession easily. Some accessions displayed significant heterogeneity and uniformity was assessed as a ‘criterion’ for marketability. However, heterogeneity was not a reason for rejection per se and if a morphotype within a heterogeneous accession was thought to have potential it was assessed.

"The trials of chicory by P 11, and the trial of Lambs Lettuce by P 10, are reported as using “one replicate block”. The terminology is wrong. The correct term is “one unreplicated trial”. (If there was only one block, there were no replicates.)".

Recommendation: the coordinator shall use the correct term in the revised report

The reviewer is correct: the text has been revised accordingly.

"The report states (WP5 task 4 Dissemination) that all partners have tried to show the progress of the project to the general and professional audience. Once again, the methodology is not described. (Did they have put posters about the project around the tasting venue? Have they got photos of the event? If so, why are these not on the web site? Have they got videos of the event?? If so, why are these not on You Tube http://www.youtube.com/? And if they have not got either videos or photos of the event, then why not?)".

Recommendation: the coordinator shall clarify in the revised report the methodology used for organising the public tasting days in organic food stores.

We thank the reviewer for the suggestion of putting photos of the tasting events on the website and we will do this. In the meantime a single photo of the event at P08 (HDRA) has been put in to the revised report. To illustrate the tasting day and results have been put in to show the commenst from the farmers working with P07 (Arche Noah).

5. WP5 – Effective management of the project

Task 1: Organisation of project and WP meetings

"In the two last annual reports, it is stated that:"- …meetings were organized . . .

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- Both meetings were attended by most partners and constructive discussions took place. - Also the atmosphere at the meetings was good."

These few words are all that are provided on all the meetings in two different years. Providing quasi-identical reports of meetings that occurred at different stages of the project, with different agendas, different objectives, and different problems to discuss perhaps even different audiences, is simply not acceptable

(Were the observers from Plantum spinach, Enza Zaden, Nickerson-Zwaan, Syngenta, and Elsoms Seeds present at either meeting? What did they say?? And who was there from ECPGR?? What did they contribute???)".

Recommendation: the coordinator shall provide the complete minutes of the third WP meeting and of the third annual project meeting as annexes of the revised report. The mention to the link to the coordinator's web site is not sufficient.

The complete minutes of the WP and third annual meeting have been attached to the revised third annual technical report as annex 4. Indeed both meetings were attended by most partners, constructive discussions took place and the atmosphere was good. As can been observed from the minutes of the annual meeting in Cesena; all action points of the previous annual (held in Alnarp) and WP (held in Schiltern) meetings were implemented, which indicates that partners are doing their job. An observer from Elsoms Seeds was present at the meeting and contributed to the discussions. People from ECPGR were present at the meetings as most/all genebankers present are members of ECPGR. For more detailed information one can consult annex 4.

Task 2: Development and implementation website

"Regarding the website, last year’s Evaluation said . . . However, there is not an external site open to the general public . . . .This criticism appears to have been acted upon in the year under review. Why are we not told? Instead the coordinator just provides 14 formulaic words (The Leafy Veg website was updated regularly and functions as a platform for the consortium.). Actually, I think quite a lot of work has been done behind the scenes, and the site is rather good. This makes the coordinator’s reticence even more difficult to understand".

Recommendation: the coordinator shall provide details concerning the progress of task 2.

We thank the reviewer for the positive words concerning the Leafy Vegetable website. To the text already present in the technical report a new text has been added. ‘In this respect the section on project highlights has been updated with a. highlights of the second project year and b. new publications. But most importantly a section on databases has been added, which represents a major step forward in the genebanking activities with respect to these crops. It should be noted that the same lay-out is used in all databases which makes a search for germplasm easier.’

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Task 3: Promote information flow between partners and contacts with the Commission

Task 4: Promote dissemination of project results

Recommendation: the coordinator shall review the text concerning tasks 3 and 4. Compared to the 2nd annual report, the third report is using the same text by using "cut and paste", just slightly modified, which is not acceptable.

Basically the text as it is, is exactly what is happening. What has been added to the original text is more detailed information on the frequency of e-reporting and contacts with the Commission. Furthermore information is given concerning the annual and WP coordinator meetings and in this context the reader is also referred to annex 4 where the minutes of both meetings are presented.

"Concerning task 4, the reports mention that: "All partners have tried to show the progress of the project to the general and professional audience".

It would have been helpful to provide a table, showing what each partner had done".

Recommendation: the coordinator shall clarify in the revised report the methodology used for task 4 (see the above comments concerning the dissemination activities for WP4-task 3).

A table has been added to the report which shows the various dissemination activities per partner in 2009.

6. Summary assessment of the work packages of the action - Overall methodology(ies)

"Because I cannot find a description of the methodologies, I am unable to comment on them".

Recommendation: the coordinator shall provide in the revised report details of the methodologies.

In the revised report the methologies used have been outlined more clearly: either by writing down the methologies used if this concerns a not yet presented methology or by referring to previous annual technical reports wwhere these methologies are described.

7. Progress in the implementation – Overview of milestones (p 36-39 of the report)

"M09: Safety duplication of part of the collections realized. It would be helpful if the coordinator marked in his table what has been achieved this year.

The numbers which have been safety duplicated by the partners, 174 accessions, have been included in the report.

M10: Start made identifying gaps in the collections. The table states that M10 is already achieved I disagree. I do not think that this has started yet.

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We do not understand where this comment of the reviewer is based on. A start of the gap analysis was made already last year and the first preliminary results for lettuce and minor leafy vegetables were presented during the annual project meeting last year (see presentations of P00 and P03 that are available on the project website). We also reported the start of this activity in the workpackage description.

M19: 500 accessions of lettuce characterized. The table states that M19 is already achieved. It would be helpful if the coordinator marked what has been achieved this year.

For Task 4 a table was included which shows the number of characterised accessions in 2009 (see above).

M30: 210 accessions of spinach evaluated. The table states that M30 is already achieved. It would be helpful if the coordinator marked what has been achieved this year.

Actually, the correct figure is 263 accessions which are ongoing, i.e. all of those accessions have already tested but only on 4 Perenospora farinosa races on a total of 7 Pfs races.

M38: 40 accessions of lettuce evaluated for utilisation and marketing. The table states that M38 already started but not yet finished; I agree. It would be helpful if the coordinator marked what has been achieved this year.

In WP4 task 3 the achievements concerning the evaluation and marketing of lettuce has been described. In total 15 acccessions of lettuce were evaluated this year by P07 Arche Noah.

M50 and 51: the table refers to" the overall progress of the project (mid-term review)" considered as achieved. I disagree. Where is the mid-term review?

In the annual meeting in Cesena a discussion took place on this subject. The general outcome of this discussion was that the project is on track and runs smoothly because all milestones and deliverables have been reached in time or will be reached in time (based on the current progress). Moreover all the progress in the last years has been scrupulously reviewed by external referees. Therefore it was thought that the writing of a mid-term review would not really add to the evaluation of the project. Of course this would have been different if the project faced severe delays and bottlenecks.

Recommendation: the coordinator shall provide in the revised report details for milestones 09, 10, 19, 30, 38, 50 and 51.

"Concerning the progress of WP4 is fulfilling its milestones, but there is not much evidence that WP4 is meeting its objectives (To evaluate material for utilization and marketing purposes resulting in specialized and regional products, increasing the diversity of products for the consumer. This includes the evaluation for organic farming systems, which will result in products to fulfil consumer demands in this area.)... Was it perhaps unrealistic to hope for increased diversity of products for the consumer? – a proper discussion of the matter must be provided in the final report".

Recommendation: the coordinator shall consider this remark for the final report.

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Material has certainly been evaluated for utilization and marketing purposes as can been red in task 3 of WP4 of the annual technical report. Most certainly we will discuss the matter in the final report.

8. Problem(s) encountered

"If there have been no major problems, there must have been minor problems. I have already drawn attention to a couple of these minor problems. I would be interested to know how they were overcome. (How did the partners overcome problems of quarantine organism blocking exchanges of material? How are they handling non uniform, heterogeneous, accessions?? How will the partners face up to the problem of finding accessions which have significant enough unique selling point for use in marketing activities?)".

Recommendation: the coordinator shall clarify in the revised report some of the minor problems encountered.

The answers to the questions raised by the reviewer have been answered already by us when we responded to earlier questions by the reviewer. The situation in this project is simply that things are running smoothly until present and that we do not expect that we will face difficulties which will block the progress of the project..

8. Dissemination of the interim results

Web publications

"I have downloaded and read all the publications listed on p 47 of the report. I did not find, in any of the above web pages, any mention of the Leafy Veg project, nor of the Gen Res programme, nor of the European Union. I regard these omissions as disgraceful…

And it seems that each partner has done their own dissemination; I do not think that any of this work has been coordinated (centralized press releases? project photographs with texts translated into all local languages?? coordinated use of new media such as blogs, You Tube, Twitter???).

Recommendation: the coordinator shall clarify in the revised report the missing references to the Leafy Veg action 001 and GENRES Community Programme on the web publications. He shall clarify the coordination for the dissemination of interim results.

Indeed the reviewer is correct when he/she states that in the web publications, there is no mention of the GENRES-EU programme. Our fault!; the partner involved (there was only one partner who was active in this area) has been informed of this omission. However it is a pity that the reviewer did not check the scientific publication output as in all three scientific publications the EU GENRES programme has been mentioned. Concerning the coordination of the dissemination activities: on the leafy veg website the highlights of a past project year are presented. Furthermore the coordinator stimulates all partners during the annual and WP meetings to disseminate their project output.

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"Something more must be done in the final year. Nowadays all universities have a press officer, and so do most research laboratories. I suggest that the coordinator, and the work package leaders, and the task leaders, should each take contact with their local press officer to see what they suggest. The partners should then meet together (either as a web-meeting, or face-to-face) to share what they have learned and to formulate a plan for the coordinated dissemination of results during the final year".

Recommendation: the coordinator shall consider this remark for the final report.

We will consider this remark for the final year.

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