COLLABORATIVE CROP RESEARCH PROGRAM The McKnight Foundation Collaborative Crop Research Program RESEARCH AND TRAINING PROGRES REPORT: December 1, 2003– November 30, 2004 TITLE OF THE PROJECT: Strengthening the On-farm Conservation and Food Security of Andean Tubers in the Fragile Ecosystems of the Southern Peruvian Highlands PRINCIPAL INVESTIGATORS

Ramiro Ortega, Universidad Nacional de San Antonio Abad del Cusco, Perú Carlos Quiros, University of California, U.S.A. Carlos Arbizu, International Potato Center Co-PI: Willy Vargas, Luis Lizárraga, José Rosas, Lynn Kimsey, Steve Brush,

Harry Kaya, Willy Roca, and Jesús Alcazar. Executive summary Seventy two morphotypes of oca, 68 of mashua, and 29 of ulluco, from the communities, were morphologically homologated with the collection maintained by the International Potato Center (CIP). It has been revealed that only 3 morphotypes of oca, 3 of mashua, and 2 of ulluco have their homologous at the CIP’s collection, suggesting that diversity of oca, ulluco, and mashua from the communities are poorly represented by the CIP’s gene bank. Molecular markers have shown that ocas from Matinga and Poques were different. Diversity of ulluco is not as extensive as in mashua. Mashuas from Picol, Matinga, and Qqueccayoq grouped separated from Poques, Chumpi, and Sayallafaya. Cultivars moved in the seed flow have been documented. Farmers’ strategies to mitigate abiotic factors have also been documented. Traditional storage and processing have been improved. Profitability of potatoes was found to be higher than the ones in oca, ulluco, and mashua. Taxonomic identification of oca weevil has finally been clarified as Ariodistus tuberculatus Voss after remaining obscure for decades. Biology and behavior of oca weevil has been determined. The parasitic nematode Heterorhabditis was found to be highly virulent to oca weevil at larvae and pupa stages. Farmers have been using strategies developed for integrated managements of Andean tubers. A total of 113 children from four communities were trained on integrated crop management. Graduated farmers of the FFS launched the Association of Organic growers of Andean tubers. A radio program has been diffusing our findings. Two B.Sc, students defended successfully their thesis; other 6 B.Sc., and 2 M.Sc. will be graduated in 2005.

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RESEARCH PROGRESS REPORT 1. Dynamics of on-farm conservation Objective To strength the factors that contribute to a sustainable conservation and utilization of oca, ulluco, mashua and Andean potatoes without damaging environment. ACTIVITIES Documenting the cultivars/morphotypes grown by farmers within each rural community and to obtain information on their geographical distribution (activity 1) Homologues morphotypes of Andean tubers (72 of oca, 68 of mashua, and 29 of ulluco), which were visually identified by their tuber and plants characters during the previous year for the six communities (Picol, Matinga, Qqueccayoq, Poques, Chumpi, and Sayllafaya), were further homologueted with the field collection maintained by the International Potato Center (CIP) in Huancayo, Perú. Homologation was carried out in two steps. In the first one, morphotypes of the communities, and those maintained by CIP were visually compared considering both their tuber and plant characters. In the second step, morphotypes of the communities and those of CIP were planted in Huancayo, where morphological data of both tuber and plant characters were recorded according to a standardized descriptor list for oca (IPGRI/CIP, 2001), and ulluco (IPGRI/CIP, 2003), and mashua descriptor list, which are being developed by CIP. Data recorded were followed by R analysis (Ihaka and Gentlemen, 1996) at distance d=0 Only 3 morphotypes of oca from the communities, 3 of mashua, and 2 of ulluco were found to have their homologues in the Peruvian collection maintained by CIP (Table 1). This would suggest that Peruvian collection of oca, mashua, and ulluco is not well represented by the Peruvian collection of these crops maintained by CIP, particularly the material grown in the communities. Although CIP maintains a number of accessions from Cusco and Puno (Southern Peruvian Andes), that material however has not been collected by CIP, but was collected mainly by NGO’s, and National universities and later on donated to CIP, or put in trust.

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Table 1. Morphotypes of oca, mashua, and ulluco identifies both visually and by R analysis, and their homologues of Peruvian material maintained by CIP. Crop Morphs of the communities Homologues at CIP’s collection Oca M 2113

P 1535 Pq 650

ATT 5467, PICA FRU 92 OC AFV 5538, ARV 5343, COC 03 08 009a, COC 013a, COC 125, COC 172, COC 176, COC 540, MH 335, MU 025, MU 026, MU 046, UNC O 338 AAAV 5589

Mashua Q 2501 S 1 S 9

ARB 5581 AVM 5557 ARB 5242 PICA 92 MA

Ulluco M S 6

PICA NAR CUS 92OL , U 096 84 AAM 5028, ARV 5342, CLC 012, CLC 013, CLC 016, CLC 018, CLC 020, CLC 023, CLC 025, CLC 030, CLC 031, CLC 032, CLC 037, CLC 046, CLC 053, CLC 068, CLSP 012, CLSP 014, CLSP 035, CLSP 036, CLSP 042, PCP 121, PCP 126, PICA HUACUS 92C

Determining tuber seed flow throughout the ecoregion (activity 2) Tuber seed flow is a dynamic strategy consciously or unconsciously used by farmers to maintain productivity of their oca, ulluco, mashua and Andean potato crops in traditional farming systems. We surveyed at random 48 growers of oca, ulluco, mashua and Andean potatoes in the villages (12 in Poques, 9 in Chumpe, 9 in Sayllafaya, 8 in Matinga, 6 in Qqueccayoq, and 4 in Picol). Farmers, were asked the following questions: (1) how are the yield of your tuber crops?, (2) What do you do to improve the yield of your tuber crops?, (3) What are the most yielding cultivars?, (4) How long have you been growing them?, (5) What is the performance of them throughout years?, (6) What do you expect for the next harvesting season in terms of yield?, (7) are you considering another strategy to improve the yield of your tuber crops? . Generally speaking, farmer’s responses were: yield is going down as tuber’s sizes are also smaller, some cultivars are tired because degeneration is taking

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place, good quality and high yielding tuber seeds are produced in the upper part of the villages but it depends on availability of appropriate lands, most farmers were considering to renew their tuber seed to counteract low yield of their crops. Farmers, places, and strategies for tuber seed acquisition have already been identified either in neighboring communities or rural fairs, good yield is associated with evenly distribution of rain throughout the cropping season. As previously reported, tuber seed flow is carried out within, and between communities. Within communities, farmers usually use tuber seeds from their own plots. Thus, a random sample of farmers taken in the communities of Picol, Matinga and Qqueccayoq, indicated that a mixture of 5-80 arrobas of Andean potato cultivars (1 arroba=25 pounds=11.5 kilos), 1-6 of oca cultivars, 1-5 of ulluco, and 0.5-2 of mashua were stored just for planting purposes. Whereas our survey in the communities of Poques, Chumpe, and Sayllafaya, showed that 6-90 arrobas of Andean potatoes, 1-30 of ulluco, 1-20 of oca, and 0.5-5 of mashua were stored by farmers exclusively for planting. Farmers of Poques, Chumpe, and Sayllafaya who had not stored their own tuber seeds acquire them by exchange of labor, loans, buying and selling negotiations, and gifts. As far as tuber seed flow between communities is concerned, tuber seeds are moved from community to community due to the lack of fulfilling family seed needs, stock degeneration, and introduction of exotic cultivars. Tuber seed movement, as previously reported, usually takes place at harvesting and planting time. Farmers themselves or buyers transport tuber seeds on carrying animals (llama, horse, mule, donkey). We have determined the incoming tuber seed from neighbor and remote communities, and regional markets into the communities of Picol, Matinga, Qqueccayoq, Poques, Chumpe, and Sayllafaya (Fig. 1a,b). Also, it was determined the outgoing tuber seed flow from project villages to neighboring and remote communities, and regional markets (Fig. 2a,b). The main cultivars moved have been documented (Table 2). The Arc View GIS program was used for exact geographical identification of the villages.

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Table 2. Most common cultivars of Andean potato, oca, ulluco, and mashua grown in the villages, and marketed or change in rural fairs of neighboring communities. Community or rural market

Andean potato Oca Ulluco Mashua

Ccolquepata Ch’aquillo, Ccachunwaccachi, ch’irita, huacca ccallo, huayro, huayruro, k’ello huaccoto, lequecho, oque lomo, paspasunchu, peruanita, puca, q’ompis, t’ica huayro, yurac lomo, yurac q’ompis

K’ello panti, Kishuar, misitu, puca oca, yana oca, yurac, yurac kayto

Kello kaytu, muru lisas, papa lisas, puca lisas

Frutilla añu, jasuti añu, muru añu, sactampilla añu, yana barba, yana añu

Huama Bole, huayro, q’ompis, oquelomo K’ello, panti, puca panti

K’ello chuccha, muro lisas, q’ompis lisas, zanahoria lisas

Huacahuacra, Sasuti añu, puca añu

Lamay Ccachunwaccachi, chimaco, huayro, imilla, llamasenca, mact’illo, oque lomo, orco amachi, puca mact’illo, puca palta, q’ompis, runtus, yana bole, yana olones

K’ello kaytu, k’ello oca, k’ello panti, misitu, puca panti, yurac kishuar

K’ello chuccha, lima lisas, muru lisas, puca lisas, papa lisas, zanahoria lisas

Frutilla añu, jasuti añu, wacawaccra añu, zapallu añu

Lares Berundus, Cachunwaccachi, ccasapuyhuan, churillo, chu’uruspe, cuchi aca, huamanero, huayro, kusi, mact’illo, peruanita, puca bole, q’ompis, runtus, yana bole

K’ello kaytu, kishuar, meztisa, misitu, puca panti

K’ello chuccha, muru lisas, papa lisas, yurac lisas

Wacawaccra, yana añu, yawarwacac, zapallo, añu

Sont’occocha Huayro, luntus, mact’illo, olones, paspa sunchu, peruanita, q’ompis, t’ica bole, yana bole

K’ello kaytu, k’ello oca, misitu, puco chillido, puca oca, puca panti, yana oca

K’ello kaytu, muru lisas, papa lisas, puca lisas

K’ello añu, puca añu, wacawaccra, zapallo añu

Sicuani Amakaya, chaquillo, ch’asca milco, chequefuro, chichiringa, hualt’a, huaynillo, kanchillo, llamac rurun, miskila, pacus, puca unchuna, q’ompis

Misquila, puca oca Jasuti añu, k’ello añu, puca añu, wacawaccra

.

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Documenting farmers’ knowledge on handling the complex of Andean tubers (activity 3) We have continued documenting star, plant, and animal indicators as well as cabañuelas (weather in August) to forecast weather for the next planting season. Our data have shown that the indicators mentioned in our two previous reports have not changed during the last cropping season in the communities. A random survey of 73 farmers (16 of Poques, 13 of Matinga, 12 of Qqueccayoq, 12 of Chumpe, 12 of Sayllafaya, and 8 of Picol) revealed that the use of star, plant, and animal indicators including cabañuelas is a common practice to forecast weather and plan the next cropping season. Furthermore, farmers try to use as many indicators as possible to decide the planting time, the most appropriate crop and cultivar, appropriate place and plots within a muyuy, and the area to be handled by a family. We have documented for instance that if a heavy rainy season is predicted, among others, early planting will be one of the alternatives to mitigate the late blight effect. Also, if a dry season is predicted, among others, the chuqui or pinchi strategy for planting potatoes will be one of the alternatives to mitigate the lack of rain. In the chuqui or pinchi strategy, potatoes are planted by making pits of 15-20 centimeters deep with a foot plow every 35-40 centimeters apart followed by putting 2-3 small tuber seeds per pit to which a handful of either caw or sheep manure is added for fertilizing purposes. Post harvest and storage technology (activity 4) Improving the taq’ues. As indicated in our previous report, taq’ues are traditional strategies used mainly by farmers of Sayllafaya and occasionally by farmers of Qqueccayoq to store Andean tubers either for self-consumption or for planting purposes. It is of easy construction, and cost effective as resources of the community can be used. So far, it has been constructed 20 taq’ues in the village of Sayllafaya (12 in 2003, and 8 in 2004). We also repeated experiments to show the value of the improved taq’ues by storing 50 kilos of oca in each taq’ue and left from June to August. At the end of the experiment, damaged tubers were weighted both in improved taq’ues and the control ones. As in the experiments of the previous year, whereas losses in the improved taq’ues showed to be less than 5%, losses in the traditional taq’ues (control) ranged from 10 to 20 %. As a consequence of the two years results, farmers have shown their willingness to continue the utilization of the improved taq’ues to store their Andean tubers. Also, farmers of neighboring communities have been showing their willingness to use the new version of taq’ues. Improving the cahuitos. This strategy is being used to store Andean tubers either for self- consumption or for planting in the six communities. The main features of cahuitos were pointed out in our previous report. We improved the efficiency of the traditional cahuitos by making 2-3 divisions within a cahuito in order to discriminate Andean tubers for self-consumption or for planting.

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Improving the processing of Andean tubers. When there is overproduction of Andean tubers, farmers usually process their oca into kaya, ulluco into linli, and potatoes into chuño and moraya or tunta. Kaya is usually prepared by soaking oca tubers (including the ones mechanically damaged or damaged by insects) for 2-3 weeks in stream water, after which tubers are exposed to sunny days to dry up. Thus, kaya can be store for years, and is essential for food security when there is shortage of food during critical months than can take place in the villages from October to January. We improved the quality of kaya preparation by improving the wells to soak oca tubers. Thus, stone and some cement were used to construct 7 wells in the community of Sayllafaya, 3 in Picol, and 3 in Qqueccayoq. Furthermore, it was strongly recommended to farmers to use clean water to guaranty quality and hygiene of kaya. At the end the kaya have better presentation for marketing or self-consumption, cleaner and more hygienic than the traditional kaya. We conducted an experiment with 5 farmers of Picol, and 2 of Qqueccayoq. Our results indicated that about 25% of the fresh oca tubers can be converted into kaya of good quality in terms of color and flavor. The traditional way on the other hand yielded less than 25% of processed product and bad quality presentation. Chuño is prepared usually by exposing small potato tubers to frost for 2-3 nights, after which water is squeezed out from tubers and tubers pealed by tramping on them. The next step is to soak potato tubers for 2-4 weeks in stream water, after which tubers are dry out in sunlight. Thus, chuño like kaya can be store for years and used for the same purposes already mentioned. We improved the quality of chuño by using the new wells constructed with stone and cement in the villages already mentioned for routine processing of potato into chuño.

So far, we have not made attempts to improve the processing of ulluco into linli. Identification of barriers to marketing (activity 5) The analysis of market barriers was carried out based upon a sample of 77 household surveys (Table 3), gathered during the second year of the project (2003).

Table 3. Sample of household surveys by community (2003)

Community Families Sample % Picol 26 5 19% Maringa 78 16 21% Qqeccayoc 58 8 14% Chumpi 83 16 19% Poques 150 20 13% Sayllafaylla 53 12 23% Total 448 77 17%

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In the 77 household surveyed we had information about agricultural production of the season 2002-2003. From a total of 646 plots, the sample included 248 plots of potatoes and 146 plots of other Andean tuber (oca, olluca and/or mashua). The overall profitability analysis showed that the average net benefit, both at plot, and family level, was better for potatoes than for oca, olluco and mashua. Larger potato returns were found in the communities of Chumpi and Poques (Lamay), and Qqeccayoc (Taray); lowest potato returns were found in Matinga (Taray). With regard to the other tubers, average returns were found greater in the communities of Qqeccayoc (Taray) and Poques (Lamay), while the lowest average returns were observed in Picol (Taray). However, profits have been very low when families sold their whole production of oca and olluco. Thus, farmers' strategy not to sell their total production is apparently necessary to guarantee a minimum of food basket. If the same analysis was carried out by production zones (high, medium, low), we would find larger profits in the higher production zones in both micro regions (Taray and Lamay). That is not the case for the other tubers. While in Taray is the middle production zone where we observe larger profits, in Lamay is the lower production zone where the returns of olluco, oca and mashua are larger. In addition to the analysis already indicated, the team at CRIBA continued to build a data base of prices of tubers at three different market places: the Ccasccaparo market, in Cusco, the market of Calca, the Province Capital in the valley of the Urubamba river, and at the Sont´ococha market, the most traditional market in the upper highlands at the micro region of the communities of the district of Lamay. A preliminary time series analysis of these prices, assessing how competitive is the price formation of tuber through different levels of price convergence, was presented in the report of the first year of the project, and will be completed during the fourth year. Precise inventory of genetic variability by molecular markers (activity 6) a. Oca. AFLP profiles, similarity and cluster analysis A total of 248 accessions of oca (122 of Matinga and 126 of Poques) were studied using 7 AFLP primer combinations. Our work detected a total of 151 polymorphic markers. The dendrogram based on the neighbor joining method and using Jaccard’s similarity index (Fig. 3), shows two groups of oca: One group represents the majority of accessions grown in the community of Matinga, and in the other one most accessions of the village of Poques are grouped. Thus, there is a separation between the two communities. The mean similarities worked out for Poques and Matinga were 0.89 and 0.88 respectively, indicating a low genetic diversity within each community.

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Analysis of molecular variance (AMOVA) AMOVA was conducted to quantify the genetic relationship among communities by partitioning the variation between, and within communities (Table 4). Whereas variation between the two communities accounted for 10.86% of the total in a significant manner, variation within communities was found to be 89.14% of the total. The large variation within communities can be explained by the wide variation cultivars or morphotypes within each community. Table 4. Analysis of molecular variance for ocas of Matinga and Poques (n=248)

Source of Variation d.f. Sum of squares Variance

components Percentage of

variation

Between communities (Matinga and

Poques)

1 311.030 2.35318 Va 10.86

Within communities 246 4750.792 19.31216 Vb 89.14

Total 247 5061.823 21.66534

b. Ulluco From the primer screening output (150 different primer combination screened) it is evident that the marker variability of this crop is not as extensive as the one observed in mashua. On the basis of polymorphism, we selected 6 primer combinations for the clustering analysis yielding 70 polymorphic markers identified with 20 primer combinations using 5 forward primers and 20 reverse primers (Fig. 4). c. Mashua Primer Screening

We selected on the basis of polymorphism the following 8 primer combinations for the clustering analysis, which yielded 120 polymorphic, easily resolved and reproducible markers (Fig. 5).

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Data Scoring and Clustering Interpretation

The sampling expansion of molecular markers, from two to eight primer combinations essentially yielded similar results to the preliminary screening of the previous year. The clustering analysis based on 120 markers shows that there is a range of similarity from 62 to 100% among the population, which is distributed in six major clusters with a relative consistent bootstrap repetition (Fig. 6, and 7). Not duplicates were found. The majority of accessions from the communities in zone 1(Picol, matinga, and Qqueccayoq) are separated from those of zone 2 (Poques, Chumpe, and Sayllafaya), although those from Sayllafaya tend to cluster together. The cultivated accessions have similar genetic backgrounds as those of the wild accessions collected in Taray and Pisac, not far from the cultivated fields of Zone 1 and Zone 2. These accessions commonly considered wild in spite of their ability to tuberize, are certainly well within the range of genetic variation of the cultivated ones. Therefore, despite of their rustic ecological and isolated geographical conditions they are most likely escapes to cultivation. This is seen more clearly on the Neighbor Joining Tree where the point of origin of its cluster is shared with cultivated accessions from Zone 1 and Zone 2, mostly Sayllafaya (Fig. 8). The discrimination of some cultivars from zones 1 and 2 indicates limited exchange of tubers used for seed between communities located in different geographic zones. The few exceptions found, where cultivars of zones 1 and 2 fell into the same cluster, may be due to seed trading and exchange taking place at the Cusco and Calca fairs and markets. The accessions from Poq'ues and Chumpe clustered together as expected since these are neighboring communities. The accessions of Sayllafaya, also in the same zone, associated to this cluster, indicating extensive germplasm exchange by farmers of these three communities. The widest range of cultivar variability was found in Sayllafaya, which is the most isolated of the communities and major city markets (91.50 kilometers from Cusco). A similar clustering pattern is shown for the accessions from Picol and Matinga,that are probably the most different from the CIP collection. In general the range of variation of the cultivars from the communities and the wild accessions are not represented in the cultivars of the CIP germplasm collection (Fig. 8.). Most of this collection originates from other regions of Peru. No association was found between clustering by molecular markers and glucosinolate content, except by the fact that the highest content of these compounds was observed in the wild accessions. The clustering based on morphological descriptors used by Duran (2004), shows only 38% of similarity. Therefore, the population seems to be more variable phenotypically (visually) than genetically (molecular markers, 62%). Furthermore, the wild accessions are separated from the cultivated mashuas in the first cluster. This might be the result of morphological adaptability (tuber shape, tuber skin color) to adverse conditions, even though genetically they share the same genetic background with their cultivated parents. The second cluster is conformed by the cultivated population from the six villages and 2 wild accessions which means that the degree of morphological differentiation was not too noticeable in these individuals. Also 26 duplicates were found, that according to molecular markers were similar but not identical. Morphological data from the Mashua CIP Collection was not available.

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Effect and Impact. It is clear from these results, as well as from the glucosinolate analysis that mashua is a genetically variable crop. Interestingly, the widest range of variation was found for the most isolated community, Sayllafaya, which indicates that the growers value and maintain the genetic variability of this crop in an effective way. This community could serve as a reservoir of genetic variation by supplying other communities with tuber seed. Efforts for in situ hybridization could be easily channeled there, where there is already a culture for maintaining genetic variation of tuber crops. Another important conclusion was the fact that the wild accessions are likely the result of cultivation escape. Interestingly, the highest glucosinolate content was found among these accessions, which could be explained by the fact that these secondary metabolites probably defend the plants from pest and disease attack. The variation observed in these accessions could be brought back easily to the cultivated accessions by hybridization and selection. Glucosinolate determination in mashua (activity 7) Based on our results of the previous year, 14 accessions of the wild mashua (CAS), and 11 of the cultivated ones (Matinga, Poques, chumpe, Sayllafaya) were taken selectively (Table 5) to study the stability of glucosinolate through different strategies of crop management. Because of their high GSL content, the wild accessions are important genetic sources to increase the content of these compounds in cultivated mashua. Some of these accessions have twice as much content than the cultivated mashuas. Therefore, through breeding it might be possible to develop lines for different purposes. High glucosinolate lines for a possible extraction of GSL derivatives with biological activity or consumption as “functional food”. Low GSL lines with better palatability to increase the popularity of this crop among rural and urban consumers as shown by some cultivated mashuas.

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Table 5. Mashua accessions selected for environmental effect and palatability survey. Concentration in uMol/1 g. of dried tuber

Number Village OHB [ ] B [ ] MOB [ ]

TOTAL GSL

[ ]

CAS-021 Kayra 4.41040 0.00000 49.98701 54.39741

CAS-023 Kayra 0.45730 0.15392 44.56820 45.17942

CAS-019 Kayra 1.87156 0.27626 32.67798 34.82580

CAS-002 Kayra 1.48354 0.00000 32.74286 34.22640

CAS-005 Kayra 0.44852 0.00000 30.59445 31.04297

CAS-007 Kayra 0.88645 0.00000 29.64616 30.53261

CAS-053 Kayra 0.33812 0.00000 29.58377 29.92189

CAS-024 Kayra 1.14005 2.02872 25.30900 28.47777

CAS-050 Kayra 1.10236 0.06793 15.36256 16.53285

2002 Matinga 0.42440 0.23241 15.07891 15.73572

CAS-058 Kayra 0.46995 0.04921 15.10209 15.62124

9 Sayllafaya 0.35094 0.13896 14.80913 15.29903

153 Sayllafaya 0.33408 0.79925 13.53344 14.66677

CAS-018 Kayra 0.05259 13.62875 0.00000 13.68133

1001 Chumpe 0.25084 2.69875 8.14042 11.09001

75 Sayllafaya 0.27204 0.54306 9.68558 10.50067

CAS-030 Kayra 0.68667 1.65815 8.00963 10.35445

583 Poques 0.28285 2.67885 4.70692 7.66862

148 Sayllafaya 0.53266 6.08245 0.07398 6.68908

CAS-059 Kayra 0.80821 4.82183 0.09526 5.72529

CAS-032 Kayra 0.00000 5.72211 0.00000 5.72211

25 Sayllafaya 0.00000 0.00000 0.50607 0.50607

17 Sayllafaya 0.00000 0.00000 0.43351 0.43351

12 Sayllafaya 0.00000 0.00000 0.32593 0.32593

1057 Chumpe 0.00000 0.00000 0.29064 0.29064

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2. Integrated crop management Objective To, develop strategies for integrated management of oca, ulluco, and Andean potato weevils as one system. Weevil taxonomy (activity 8) A. Weevil identifications to date:

Oca weevil.

In collaboration with Charles O’Brien we have finally resolved that the correct genus and species of oca weevil is Adioristidius tuberculatus Voss. This species was identified from 70,000 specimens collected in oca fields in Cuzco by Wille Vargas and in Huancayo by Jesus Alcazar. This is a small distinctive species, with tuberculate wing covers. The generic name of this species has caused considerable confusion, with the names Microtrypes, Adioristus and Macrostyphlus being used. Examination of types in European museums and original descriptions confirm that Adioristidius is the correct name. Morrone elevated Adioristidius to genus in 1994.

Fig. 9. Adult oca weevil Larval and pupal oca weevils (left).

Ulluco weevil.

A species of Cylydrorhinus is clearly the pest attacking ulluco tubers. However, we have not yet found a described species that fits this one. It may well be an undescribed species. To date twenty species of Cylydrorhinus have been borrowed from Kuschel’s collection in New Zealand.

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B. Potential Parasitoid Wasps:

Malaise trapping in the community of Picol revealed a number of parasitoid wasps in the family Mymaridae. These are egg parasites, which may attack the eggs of oca, ulluco and potato weevil. These specimens are being sent to Serguei Triapitsyn at the University of California, Riverside for identification. Discussion Species identifications have to be confirmed by comparisons with museum species, particularly primary types. The dominant weevil species taken in oca fields is Adioristidius tuberculatus. Examination of a newly harvested oca field above the village of Picol in May 2003 revealed 8 genera of weevils, including Adioristidius, Cylydrorhinus, Premnotrypes, Amitris and others as yet unidentified. This field was also infested with larval scarabaeid beetles and cutworms (family Noctuidae). There were also parasitoids of the scarabaeid beetles; these were wasps (probably a new species) in the genus Elaphroptera (family Tiphiidae). Biology and behavior of oca and ulluco weevil (activity 9) The biology and behavior of oca weevil

It has been found that the life cycle of oca weevil lasted 348 days, eggs 22 days, larvae 59 days, pre-pupae 27 days, pupae 33 days, over wintering adult 35 days and adult 158 days (Table 6).

Host range

Our studies have shown that oca weevil can also eat a range of cultivated plants such as ulluco, mashua, potatoes, broad bean, barley and wheat (Table 7). Also, the pest can survive eating a wide range of wild species (Table 8). The female weevil can put their eggs in all the indicated plants with different rates of success. The effect of temperature Studies about the effect of temperature showed high survival rate at 5 oC with 65 % (Table 9.) and the lethal temperature was at 25 oC, in which all were dead. The major number of adults was found at 15oC with 52.5 % (Table 8). Entomopathogens fungus The fungi Beauveria bassiana, B. brongniartii, Verticillium lecani, Paecilomices sp and Beauveria sp were identified as entomopathogens on larvae and adults of oca weevil.

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Table 6. Life cycle of oca weevil Aristidius tuberculatus Voss. Cusco, 2003-2004.

Stage Average Standard desviation Minimun Maximun

Egg 22.47 12.64 8 41

Larvae I 11.7 5.69 4 23

Larvae II 11.95 4.61 5 20

Larvae III 16.49 6.13 8 35

Larvae IV 21.45 7.37 7 39

Pre-pupae 37 10.59 18 56

Pupae 33.18 9.78 14 50

Winter adult 35.5 15.87 17 61

Longevity

adult 158.5 5.46 150 167

Total cycle 348.24

Table 7. Host range evaluation of oca weevil (Aristidius tuberculatus Voss) rearing in cultivated species. Cusco, 2004

Crop Species Leaf area mm2/weevil

Eggs/female % eclosion

Oca Oxalis tubererodum 7,885.6 296 97.22

Ulluco Ullucus tuberosum 2,782.6 90.6 94.26

Mashua Tropaeolum tuberosum 2,070.5 56.2 93.59

Broad bean Faba bean 1,862.2 32.2 99.37

Potato Solanum tuberosum 1,065.3 29.2 92.46

Barley Hordeum vulgare 955.3 4 80.00

Wheat Triticum vulgare 688.5 4.6 78.26

15

Table 8. Host range evaluation of oca weevil (Aristidius tuberculatus Voss) rearing in

wild species. Cusco, 2004

Vernacular name

Species Leaf area mm2

Eggs/ female % eclosion

Salvia Salvia opposiflora 2,849.1 21.6 91.66

Kiko-kiko Bidens sp. 1,523.7 8.8 79.54

Nabo Brassica campestres 946.6 76.4 94.24

Auja-auja Erodium sp. 922.6 42 92.38

Maycha Senecio sp. 694.8 13.4 91.04

Cebadilla Bromas uniloides 638.8 4.6 82.60

Kikuyo Pennisetum clandestinum 346.4 2.6 84.61

Ñuska Astragalus garbancillus 174.3 1.6 100.00

Manca caqui Calceolaria pinnata 173.0 0.6 100.00

Table 9. Effect of temperature on the survival of oca weevil Adioristidius tuberculatus after 90 days. La Molina, 2004.

Temperature

Initial population Larvae IV

Alive larvae %

Dead larvae %

Alive

pupae %

Dead pupae %

Alive

adults %

Dead adults %

Total weevi

ls alive

%

Total weevi

ls dead

% 5 oC 40 52.5 35 12.5 0 0 0 65 35

10 oC 40 2.5 45 27.5 7.5 15 2.5 45 55

15 oC 40 0 35 0 12.5 52.5 0 52.5 47.5

18 oC 40 0 32.5 0 15 50 2.5 50 50

20 oC 40 0 27.5 0 32.5 35 5 35 65

25 oC 40 0 87.5 0 7.5 0 5 0 100

16

Integrated management of weevils (activity 10) In 2003, a Heterorhabditis the matode species referred to as Alcazar-1 was isolated from potato weevil larvae in soil from a potato storage shed in Huasahusai. From mid-June to mid-September 2004, UC Davis graduate student, Mr. Soroush Parsa, conducted research with the nematode at CIP in the laboratory of Mr. Jesus Alcazar. During this time, these accomplishments were made following the work plan established last year. The pathogenicity (i.e., virulence) of Alcazar-1 was tested against the oca weevil. Lethal dose experiments have demonstrated that Alcazar-1 is highly virulent to the oca weevil with 1.6 infective juveniles needed to kill 50% of the larvae at 20 C (lethal concentration that kills 50% = LC50). Comparison assays using a single nematode concentration per insect suggest that there is no significant difference in susceptibility between larvae and pupae. The ability of the nematode to kill weevil larvae, pupae and over wintering adults inside oca tubers has been demonstrated. Field infested ocas were submerged in a nematode suspension, and one week later, the tubers were dissected and checked for oca weevils. At least one infected weevil was found in 54% of the tubers and, on average, nematodes killed 24.7% of the weevils within a tuber. Furthermore, when infested ocas were planted under 10 cm of sand and the nematodes applied on top at a concentration of 25 infective nematodes/cm2, all ocas had infected weevils after two weeks. The average percent mortality was 63.1%. Infective juvenile emergence per oca weevil larvae cadaver was estimated at 11,884 ± 2,530 infective juveniles. This amount corresponds to an average of 619,905 ± 116, 948 infective juveniles produced per gram of oca weevil larva. Accordingly, oca tuber seeds treated with nematodes can be a source of nematode inoculum with the potential to suppress weevils in the field. Further surveys conducted have revealed at least one putative entomopathogenic nematode isolate (Steinernema sp.) from a soil sample from the community of Sayllafaya. This nematode was isolated using the waxworm baiting technique. Progress for integrated management Weevil management strategy is based on the wide knowledge of the biology, identification of infestation sources, and development and validation of new strategies. Thus strategies are oriented to reduce the over-wintering population from infestation sources, and to control the weevil in the field through several control alternatives with emphasis on cultural, mechanic and biological measures such as destruction of crop residues, winter plowing after harvest, use sheets at harvest to prevent larvae from pupae, winter plowing in sorting and storage areas, use chickens as predators, use fungus and repellents plants in storage areas, use healthy seed, use of oca sprouts, eliminate volunteer plants, put ash at plant base, high hilling, and early harvest. Farmers of Picol, Matinga, Qqueccayoc, Ch’umpe, Poques, and Sayllafaya have being using these strategies to

17

prevent their Andean tubers from the pests. Thus, in the communities of Picol, Matinga and Qqueccayoc, 30 farmers and 40 children carried out the plowing of the infestation sources in warehouses and complemented the control by using chickens to predate the weevil larvae. Likewise, in the communities of Ch’umpe, Poques and Sayllafaya, 70 farmers and their children carried out the plowing of the sources of infestation and used chickens to predate the weevil larvae.

3. Diffusion of knowledge and training to promote agro-biodiversity conservation and food security (activity 11)

a. School children. School’s teachers, parents, and project participants agreed to complement education of children of their respective communities. Thus, 113 school children (43 of Poques and Chumpe, 40 of Matinga, and 30 of Sayllafaya) were trained at planting and harvesting time (2 months) on identification of the main pests of theirs Andean tuber crops, biology of the pests, use of manure, identification and utilization of use of repellent plants, and special care for rooting sprouts. We have been feeling the change on children’s attitude towards a systematic resources utilization of their communities.

b. Farmer field Schools (FFS). During different stages of the Andean tuber crop

development, 91 farmers (45 of Matinga, 26 of Qqueccayoq, and 20 of Picol) volunteer to participate in FFS of each village, where 16 meetings were held per village. Topics treated were identification of constraints for integrated crop management, and strategies to tackle the problems. Some farmers could not finish the FFS, but those who were able to attend continuously to the end, were examined about their knowledge acquired in the FFS. Less than 50% of the farmers passed their examinations (18 in Qqueccayoq, 17 in Matinga, and 9 in Picol), and were graduated as alumni of the FFS. Furthermore, All farmers who graduated, and to which other farmers volunteered to join giving as a result 46 farmers (18 of Matinga, 18 of Qqueccayoq, and 12 of Picol) decided to launch the Association of Organic Growers of Andean Tubers. The main objective was to have comparative advantages once farmers market their Andean tubers in the touristic city of Cusco. The project, of course, has supported this initiative by complementing farmers’ initiative.

c. Farmer’s workshop. Three workshops were held in the rural communities (2 in

Matinga, and 1 in Sayllafaya) to strengthen sustainability of our results. The first one was a farmer to farmer workshop, and took place in the village of Matinga on 1-2 October 2004 where 43 farmers of Matinga itself, and those of Picol, Qqueccayoq, Chumpe and Sayllafaya attended the event on integrated management of Andean tuber crops with emphasis on identification of the Andean tuber pests, biology and behavior of the pests, host range, and strategies either o prevent or control the pests.

18

The second was also a farmer to farmer workshop held in the village of Matinga on 11 October 2004. Leading farmers of Matinga interacted with 33 authorities and leaders of neighboring communities such as Llaq'uepata, Rayanniyoc, Ccaccacocllo, and Ch'ita Pampa. The topics treated were the strategies developed by the Andean tuber project for efficient management of tuber crops such as cultivar identification, sprouts handling, use of organic fertilizers, utilization of repellent plants and the entomopathogenic Beauveria bassiana for weevil management.

A third workshop was held in the village of Sayllafaya on 15 october 2004. As in the former case, 101 leading farmers of Sayllafaya and neighboring communities of Chumpe, Huarqui, Huanco Mullmuncos, Huata, Sayhua, Janac Chuquibamba, Juchuy Qosqo, Chuqibamba, Huancco, and Inquilpata participated in the meeting. Although the topic treated were almost the same as in the former workshop, post harvest management and Andean tubers for self-consumption and planting purposes were also treated.

d. Broadcasting. The diffusion of our findings has continued by means of a radio

program broadcasted by INTIRAYME Radio 830 AM every Saturday from .4.00 to .5.00 a.m. since December 2003. The program is in Quechua (Inca languaje) and is being addressed to Andean farmers, particularly to the six villages of Cusco, and their neighbors.

e. Participation in congresses.

Eighteen project participants, technicians, lecturers of Cusco University, and thesis students participated in the XLVI Peruvian Convention of Entomology held in Arequipa, Peru from 7 to 11 November 2004. Three project participants, and three students presented their findings. The others presented posters. The main papers presented were: “Potential of the native entomopathogenic Heterorhabditis sp. for Andean weevil control”, “collection, multiplication and preliminary pathogenic tests of entomopathogenic nematodes on different pests of Cusco”, “natural enemies of Andean weevils, utilization of the fungus Beauveria bassiana in oca stores for control of Microtypes sp.”

f. Seminars.

1. Title: Situación actual de la taxonomia de los gorgojos de tubérculos

andinos (Current status of Andean tuber weevils taxonomy), held at Cusco University on 20 march 2004

Speaker: Dr. Lynn Kimsey

2. Title: Morfotipos de tubérculos andinos mantenidos en seis comunidades

del Cusco (Morphotypes of Andean tubers maintained in six communities of Cusco), held at Cusco University on 30 March, 2004

19

Speaker: Dr. Carlos Arbizu

3. Title: Fortalecimiento de la conservación in situ y la seguridad alimentaria

de los tubérculos andinos en los ecosistemas frágiles de los Andes altos del Sur del Perú (Strengthening the On-farm Conservation and Food Security of Andean Tubers in the Fragile Ecosystems of the Southern Peruvian Highlands), held at Technological University of Los Andes, Abancay, Peru.

Speaker: Ir. Ramiro Ortega

g. Students training in Peru. Two B.Sc. agronomy students defended successfully

their thesis during 2004, six have been conducting their research, and 2 M.Sc. have also being doing their research thesis as following:

g1. B.Sc. Defended their thesis (graduated as agronomists) 1. Name: Rodolfo Viguria (Mr.)

Title of the thesis: Diagnostico de plagas en tubérculos andinos (oca, ulluco, mashua y papas nativas) en las comunidades campesinas de Chumpi y P’oques (Diagnostic of pests on Andean tubers (oca, ulluco, mashua, and Andean Potato) in the peasant communities of Chumpi, and P’oques).

2. Name: Leoncio Cusihuallpa (Mr.)

Title of the thesis: Diagnostico de plagas en tubérculos andinos (oca, ulluco, mashua y papas nativas) en la comunidad campesina de Sayllafaya (Diagnostic of pests on Andean tubers (oca, ulluco, mashua, and Andean Potato) in the peasant community of Sayallafaya).

g2. B.Sc. (Thesis in progress) 1. Estenio Durand (Mr.)

Title of the thesis: Caracterización morfológica de oca, ulluco, mashua y papas cultivadas en seis comunidades del Cusco (Morphological characterization of oca, ulluco, mashua, and Andean potatoes grown in six villages of Cusco)

Expected date of defence: February, 2005

2. Name: Karina Vargas (Miss)

20

Title of the thesis: Colección, aislamiento, identificación y patogenicidad del hongo blanco en el gorgojo de la oca (Collection, isolation, identification, and pathogenicity of the white fungus in oca).

Expected date of defence: March, 2005

3. Name: Paulo Cesar Quispe (Mr.)

Title of the thesis: Plantas cultivadas y silvestres como hospederos del gorgojo de la oca (Wild and cultivated plants as host of the oca weevil)

Expected date of defence: April, 2005

4. Name: Henry Bearnaola (MR.) Title of the thesis: Diagnóstico rural participativo de las comunidades de Chumpe y Sayllafaya del distrito de Lamay, provincia de Calca (Rural participatory diagnostic of the communities of Chumpe and Sayllafaya, Lamay, Calca) Expected date of defence: June 2005

5. Yanina Usucachi (Miss)

Title of the thesis: Evaluación de productos orgánicos en el control del waytu de la papa (Stenoptycha coelodactyla) en la comunidad de Matinga (Evaluation of organic products on the control of waytu of Potato (Stenoptycha coelodactyla) in the community of Matinga

Expected date of defence: August, 2005

6. Name: Hector Ortiz (Mr.)

Title of the thesis: Caracterización morfológica de las papas natives manejadas por los agricultures de las comunidades de Picol, Matinga, Qqueccayoq, Chumpe, Poques, and Sayllafaya (Morphological characterization of Andean potatoes grown by farmers of the communities of Picol, Matinga, Qqueccayoq, Chumpe, Poques, and Sayllafaya. Expected date of defence: September, 2005

g3. M.Sc.

1. Name: Antonio Vargas (Mr.)

21

Title of the thesis: Estudio del ciclo biologico del gorgojo de la oca (Ariodistus tuberculatus Voss Coleoptera: Curculionidae) (Studies on the biology of oca weevil, Ariodistus tuberculatus Voss Coleoptera: Curculionidae)

Expected day of defence: June 2005

2. Name: Oscar Ortega (Mr.)

Title of the thesis: Genetic variability inventory mashua by molecular markers and glucosinolate determination. Expected date of defence: June, 2005

22

References

Durand, E. 2004. Caracterización morfológica de oca, ulluco, mashua y papas cultivadas en seis comunidades del Cusco (Morphological characterization of oca, ulluco, mashua, and Andean potatoes grown in six villages of Cusco) (Thesis in progress) Ihaka, R. and R. Genthemen. 1996. R: a language for data analysis and graphics. Journal of Computational and Graphical Statistics. 5: 299-414 IPGRI/CIP. 2001. Descriptores de oca (Oxalis tuberosa Mol.). Instituto Internacional de Recursos Fitogenéticos, Roma, Italia; Centro internacional de la Papa, Lima, Perú IPGRI/CIP. 2003. Descriptores del ulluco (Ullucus tuberosus). Instituto Internacional de Recursos Fitogenéticos, Roma, Italia; Centro internacional de la Papa, Lima, Perú Morrone, J. J. 1994. Systematics, cladistics and biogeography of the Andean weevil genera Macrostyphlus, Adioristidius, Puranius and Amathynetoides, new genus. American Museum Novitates 3104:1-64.

23

Fig. 1a. Incoming tuber seed of oca (red arrow), ulluco (yellow arrow), and Andean potatoes (green arrow) into the villages of Picol, Matinga, and Qqueccayoq

Communities that provide tuber seed to Picol: Poques, Chumpe, Huaccoto, Cuyo Grande, Chinchero, Andahuaylas.

Communities that provide tuber seed to Matinga: Poques, Chumpe, Cuyo Grande, Sipascancha Alta, Pampallacta, Chinchero, and Andahuaylas

Communities that provide tuber seed to Queccayoq: Huaccoto, Cuyo Grande, Paucartambo, Andahuaylas.

Fig. 1b. Incoming tuber seed of oca (red arrow), ulluco (yellow arrow), mashua (blue arrow), and Andean potatoes (green arrow) into the villages of Poques,

Chumpe, and Sayllafaya

Providers of tuber seed to Poques: Communities of Sayllafaya, Chumpe, Ttio, Huata, Sayhua, Huancco, Huarqui, Tocra, Colquepata, Challabamba, Calca, and Amparaes.

Providers of tuber seed to Chumpe: Communities of Poques, Sayllafaya, Huancco, Ttio, Sayhua, Ccachin, Pampallacta, Huata, Huarqui, Challabamba, Chinchero, Calca, and Cusco market.

Providers of tuber seed to Sayllafaya: Communities of Carniceros, T’ocra, Ttio, Sipascancha Alta, Huata, Chumpe, Poques, and Calca.

Fig. 2a. Outgoing tuber seed of oca (red arrow), and Andean potatoes (green arrow) from the villages of Picol, Matinga, and Qqueccayoq to neighbouring communities, and regional markets

Outgoing tuber seed from the community of Picol to the villages of Matinga and Llaquepata, and the market of Cusco.

Outgoing tuber seed from the community of Qqueccayoq to the villages of Picol,

Matinga, Huata, Huancalle, Llaquepata, and the market of Cusco.

Fig. 2b. Outgoing tuber seed of oca (red arrow), ulluco (yellow arrow), mashua (blue arrow), and Andean potatoes (green arrow) to neighbouring

communities, and regional markets

Outgoing tuber seed from the community of Poques to the villages of Huarq’ui, Huancco, Sayhua, Huata, Huayllabamba, Ttio, Sayllafaya, Chumpe, and Llanchu

Outgoing tuber seed from the community of Chumpe to the villages of Chuquibambilla, Huancco, Sayhua, Huata, Pampallacta, Llanchu, and Chinchero.

Outgoing tuber seed from the community of Sayllafaya to the villages of Cachibamba, Kurpo, Huarqui, and Pampallacta.

Figure 3. Neighbor joining dendrogram of 248 accessiosn of oca based on Jaccard’s similarity index

Fig. 4: Primer screening in 6 accessions of ulluco using 6 different

SRAP primer combinations. In bold, primer pairs selected for the study.

ODD16 SA12 (700)

BG14 EM2 (700)

ODD11 EM1 (700)

CE2 SA12 (700)

ODD20 SA14 (700)

FA1 EM1 (700)

Fig. 5. Primer screening in 12 accessions of mashua using 4 different SRAP primer

combinations. In bold, primer pairs selected for the study.

Figure 6. SRAPs amplified using the ODD9-SA12 (700) primer combination in 46 mashua accessions.

Arrows show 22 polymorphic markers

Figure 7: Tree matrix of nested clusters obtained according to accession similarity

levels based on SRAP markers in Mashua

C3100

C3099

C3098

C3097

C3074

C3101

C3096

C3095

C3065

C3045

C3043

C3042

C3040

C3038

C3039

C3020

C3050

W45

W37

W36

W30

C3018

S94S22S144S19S7 S138C

H1039

S149S148S69S67S68S66S64W

53W

43W

41W

42W

40W

39W

38C

3126C

3022C

3021W

5S145S1 S121S96W

1C

H1019

PQ

502W

60W

59W

58W

57W

54W

63W

50W

49W

20C

3044M

2020P1515M

2004P1511P1509S92S89Q

2501M

2017S81S23M

2016PQ

504W

32W

21W

23W

19W

24W

18W

11W

10PQ

584W

64W

9W

8S83C

H1010

M2013

S85S79S65S62S54S130W

29W

26S2 S153S150S77S135S106S13C

H1052

PQ590

PQ586

W7

CH

1030S80S10C

H1004

PQ547

PQ588

CH

1053Q

2502W

2S4 PQ

534P1508PQ

525S82C

H1057

W61

PQ570

CH

1054C

H1001

M2001

S90S47S34P1517PQ

532C

H1055

CH

1026C

H1017

M2014

M2010

M2009

CH

1020P1506M

2015M

2008PQ

573M

2002S11PQ

529W

22P1504P1502

Huancavelica Pun Cusc

CULTIVATED - WIL

Sayllafay Pisac Cus Saylla

ZONE 2 - WILD - CULTIVATED CIP (CUSCO)

Taray Acos Zone 1 Zone 1 and

Z 2

Wild

WILDSayllafa Sayllafa

MOSTLY SAYLLAFAYA

Zone 2

MOSTLY ZONE Zone 2 Zone

MOSTLY

1 2 3 4 5 6

0.60

0.65 0.70

0.75

0.80

0.85

0.90

0.95

1.00

Coefficient

Figure 8. Un-rooted tree generated by Neighbor Joining Analysis

CULTIVATED SAMPLES - CIP

WILD ACCESSIONS