poster58: Development and implementation of methodologies for the production of clean, root rot tolerant avocado plants

DEVELOPMENT AND IMPLEMENTATION OF METHODOLOGIES FOR THE PRODUCTION OF CLEAN, ROOT ROT TOLERANT AVOCADO PLANTS CLEAN A. Mejía Jiménez1, J.P. Villamizar1, M.L. Orozco1,2, A. Arenas1, E. Alvarez1, M. Rodríguez2, B. J. Orozco2, J.A. Carmona3, D. Rios3, J. Jaramillo2 and A. González1 (1)Centro Internacional de Agricultura Tropical CIAT, A.A 6713, Cali, Colombia; (2)Corporación Colombiana de CIAT A A 6713 Cali Investigación Agropecuaria (CORPOICA), Palmira, Colombia; (3) Profrutales LTDA, Candelaria-Valle, Colombia Introduction I t d ti Outputs of this project: A FIG. 1 Collection of avocado accessions. Phytophthora cinnamomi, the agent cinnamomi, Fig. 1A. Identification of • The nurse seed-etiolation-double-grafting seed-etiolation-double- “escape trees” growing responsible for root rot, is the most rot, methodology (Frohlich and Platt 1971; Brokaw (Frohlich vigorously on plantations limiting disease in avocado (Persea 1976) was adapted and implemented in local devastated by root rot, as a source of tolerant americana) americana) around the world. world. nurseries. i avocado clones; Figs. Colombia’s avocado industry is • 254 accessions of local avocado germplasm 1B, C, E, F, G, H, J, K, and L variability of fruit developing, but faces the challenge of were collected and sub-set tested against local sub- B C D forms and shapes of dealing with under-developed under- strains of P. cinnamomi. cinnamomi. some of the 254 avocado accessions technologies and practices that • Collected germplasm represents a wide range of collected in 16 resulted in thousands of hectares altitudinal conditions (5 to 2183 m.a.s.l) m.a.s.l m a s l) l) E F Colombian departments between, 5 and 2183 affected by the disease. • 30 accessions were collected as escapes from P. H I m.a.s.l.; Figs. 1D tree cinnamomi infested soils. G trunk circumference (3.4 Tolerant Rootstocks must be used to • 21 nurseries trained on methodologies for clonal m) of one of the J collected trees. protect large investments in avocado and pathogen free propagation of rootstocks. L K farms. • A new methodological approach t t t avocado th d l i l h to test d Some facts: germplasm produced reliable information. • Collected material was incorporated into the • P. cinnamomi cause losses of 30-50% (Tamayo 30- Colombian Avocado Germplasm Bank in Fig. 2 Isolation and characterization of the pathogen causing root 2007) in commercial nurseries. Corpoica- Corpoica-Palmira rot of avocado in Colombia. Fig. 2A, root samples were taken from dying root rot suspected trees in 9 Colombian departments; isolation • The most eco-efficient strategy to alleviate eco- • Basic and inexpensive tissue culture techniques and characterization of the pathogen morphologically (Fig. 2B) and the problem is the use of tolerant rootstocks were used to produce clean avocado mother molecularly through ITS amplification and restriction (Drehnt et al. 2006; (Drehnt • Root rot tolerant rootstocks have been plants for clonal propagation FIG 2C); Fig. 2D, study of the pathogenicity of the P. cinnamomi isolates through innoculation of Hass; Fig. 2E, places from where P. developed for temperate but not for tropical • PCR analysis with Phytophthora specific primers cinnamomi isolates were obtained climates (Drenth et al. 2006) showed that disinfested Bolívar Cesar • Root rot to e a t rootstocks must be p opagated oot ot tolerant ootstoc s ust propagated microshoots recovered from escapes growing in Text Carmen de Bolívar Manaur e A Antioquia E vegetatively P. cinnamomi infested soils are Phytophthora free La Ceja Carmen de Santander (Fig. 3J) Viboral Nariño Landazuri San Vicente Fig. 3 Development of methodologies for producing • Inoculation of P. cinnamomi confirmed the El Retiro Caldas pathogen free avocado clones. Figs. 3A, B and C, method for Anserma disinfection of buds (microshoots) recovered from “escape (microshoots) susceptible nature of cultivar Hass (Fig. 2D) and Quindío Pijao Palestina Pensilvania trees” in root rot infested plots, with ethanol and sodium to e a t atu e of G755 and u e tolerant nature o G 55 a d Duke 7. B Pueblo Valle V ll hypochlorite under vacuum ; Fig. 3D, innitial in vitro culture Argelia Nuevo surrounded by an antibiotic and antimycotic solution; Fig. 3E • Out of 55 Phytophthora isolates, 49 were of P. Candelaria El Cerrito Tolima dentification of still contaminated microshoots; Fig. 3G, microshoots; C Alvarado production of new buds 4 weeks after starting in vitro culture; cinnamomi, cinnamomi, and 6 of two unidentified QuickTime™ and a decompressor are needed to see this picture. Palmira Mariquita Figs. 3H and I, plant recovery after grafting of disinfested Phytophthora species (see poster by Alvarez et D Cauca Cajibio microshoots onto seedling rootstocks in the greenhouse; Fig. al.). Isolates were obtained from root samples of Popayán Cauca 3J, PCR analysis with Phytophthora specific primers (Drenth et (Drenth al. al 2006) showing that disinfested microshoots recovered from dying avocado p y g plants Control ANR-017 CAA-103 CAT-049 Popayán Tambo T b “escapes trees” are Phytophthora free. Fig. 4 Adaptation of the nurse seed-etiolation-double-grafting seed-etiolation-double- A B C methodology (Frohlich and Platt 1971; Brokaw 1976). Fig. 4A, buds of the potential clonal rootstocks grafted onto “nurse seeds” Conclusions: (Figs. 4B and C); Fig. 4D “etiolation” culture of the grafted shoots in the darkness for improving rooting capability (Fig. 4J); Fig. I and J • Phytophthora cinnamomi is widely dispersed and plant of a commercial cultivars (Hass) grafted on a cloned root rot represents a serious threat to plantations in Colombia tolerant rootstock (G755) produced and planted at CIAT. • Methodologies developed for disinfection and in vitro A B C culture are useful for establishing clean clonal stocks D E G • Use of clonally propagated, root rot tolerant rootstocks is recommended • G755 and Duke 7, or cloned “escapes” are good candidate rootstocks for Colombia H I D E F G • The adapted methodology for production of clonal rootstocks can be applied in a cost effective way to produce root rot tolerant plants Acknowledgments This project was supported by the Colombian Ministry of Agriculture and Rural Development (MADR) and the Fondo Nacional de Fomento H I J Hortofrutícola (FNFH). P. cinnamomi Negative Bibliography AVOCADO ESCAPES AND CLONES Strains Control Brokaw WH (1976). Method of rootstock propagation J http://www.google.com/patents?vid=USPAT4012866&id=UtErAAAAEBAJ&dq=brokaw+avocado+propagation M 1 2 3 4 5 6 7 8 9 10 11 12 13 14 M Drenth A., G. Wagels, B. Sendall, C.O´Dwyer, G. Irvine and and J.A.G Irwin (2006). Development of a DNA-based method for detection and identification of Phytophthora species. Australian Plant Pathology Society 35,147-159. Durán-Barón R, y E. Daza (2003). Estado actual del sistema de producción aguacate en la costa Caribe colombiana PERSEA FOCUS 2. http://www.colpos.mx/ifit/aguacate2/espanol2/colombiaesp.htm Frolich and Platt (1972) Use of the etiolation technique in rooting avocado cuttings. California Avocado Society 1971-72 Yearbook 55: 97 109 http://www avocadosource com/CAS Yearbooks/CAS 55 1971/CAS 1971 72 PG 097 109 pdf 97-109 http://www.avocadosource.com/CAS_Yearbooks/CAS_55_1971/CAS_1971-72_PG_097-109.pdf Lozano T., J. C. 2004. La muerte precoz de árboles de aguacate en Colombia. Ascolfi Informa Vol. 30, No 5 Septiembre- Octubre. Tamayo, J. M. 2007. Enfermedades del Aguacate. Politécnica 4, 51-7

poster58: Development and implementation of methodologies for the production of clean, root rot tolerant avocado plants

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