Abstract
Crown blight, caused by Phytophthora capsici, is the most important disease of pepper (Capsicum annuum) in the world and causes great economic losses in Costa Rica. Alternatives to chemical control against this disease are crucial to prevent damage to human health and the environment. The antagonism of Plant-Growth-Promoting Rhizobacteria (PGPR) on P. capsici, and its ability to reduce wilt in pepper plants were evaluated. PGPR strains previously isolated from sugarcane and rice were identified, using 16S RNA gene sequence, as Pseudomonas fluorescens PC4, Stenotrophomonas rhizophila PC9, Pseudomonas fragi PC11 and Azospirillum lipoferum PCJ2. The inhibition of P. capcisi growth was evaluated in vitro, in the presence of the PGPR. The effect of the four bacterial strains on pepper plants inoculated with P. capsici (100 zoospores.plant-1) was evaluated in the greenhouse. P. fluorescens PC4, S. rhizophila PC9 and A. lipoferum PCJ2, inhibited in vitro growth of the oomycete by 54%, 30% and 50 % respectively, while S. rhizophila PC9 increased by 14% shoot fresh weight of pepper plants at the greenhouse. Furthermore, PCJ2 and PC9 strains reduced the shoot and root severity of the disease, and PCJ2-inoculated plants showed no symptoms at all, indicating that PC9 and PCJ2 are promising rizobacteria for the control of crown blight in pepper.References
ALTSCHUL S.F., MADDEN T.L., SCHAFFER A.A., ZHANG J., ZHANG Z., MILLER W., LIPMAN D.J. 1997. Gapped BLAST and PSI- BLAST: a new generation of protein database search programs. Nucleic Acid Research 25(17):3389-3402.
AMORIM E.D.R., MELO I.D. 2002. Ação antagônica de rizobactérias contra Phytophthora parasitica e P. citrophthora e seu efeito no desenvolvimento de plântulas de citros. Revista Brasileira de Friticultura 24(2):565-568.
BASHAN Y., de-BASHAN L.E. 2010. Chapter 2. How the plant growth-promoting bacteria Azospirillum promotes plant growth-A critical assessment. Advances in Agronomy 108:77-136.
BASHAN Y., de-BASHAN L.E., PRABHU S.R., HERNANDEZ J.P. 2014. Advances in plant growth-promoting bacterial inoculant technology: Formulations and practical perspectives (1998– 2013). Plant and Soil 378(1-2):1-33.
BASHIRZADEH Z., SHARIFI R.S., SEDGHI M., KHAVAZI K. 2012. Evaluation of rate and seedfilling period of barley to co-inoculation with plant growth promoting bacteria (PGPR) and nitrogen fertilizer. Research Journal of Agricultural Science 44(3):3-8.
BHATTACHARYYA P.N., JHA D.K. 2012. Plant growthpromoting rhizobacteria (PGPR): emergence in agriculture. World Journal of Microbiology and Biotechnology 28(4):1327-1350.
BRUNEL B., GIVAUDAN A., LANOIS A., AKHURST R.J., BOEMARE N. 1996. Fast and accurate identification of Xenorhabdus and Photorhabdus species by restriction analysis of PCR-amplified 16S rRNA genes. Applied Environmental Microbiology 63(2):574-580.
CORRALES O., VARGAS E., MOREIRA M.A. 1990. Efecto de la materia orgánica en el combate de la pudrición basal del chile dulce (Capsicum annuum) causada por Phytophthora capcisi. Agronomía Costarricense 14(1):9-14.
EZZIYYANI M., PÉREZ C., REQUENA M.E., AHMED A.S., CASTILLO M.E.C. 2004. Evaluación del biocontrol de Phytophthora capsici en pimiento (Capsicum annuum L.) por tratamiento con Burkholderia cepacia. Anales de Biología 26:61-68.
FONTECHA G. 2003. Análisis comparativo de las comunidades de procariotas intestinales de Rothschildia lebeau (Lepidoptera) mediante los polimorfismos en los fragmentos terminales de restricción (T-RFLP) del gen ARNr 16S. Tesis de maestría, Universidad de Costa Rica, San José, Costa Rica. 139 p.
GUO J.H., QI H.Y., GUO Y.H., GE H.L., GONG L.Y., ZHANG L.X., SUN P.H. 2004. Biocontrol of tomato wilt by plant growth-promoting rhizobacteria. Biological Control 29(1):66-72.
HALL T. 1999. BioEdit: A user- friendly biological sequence alignment editor and analysis program for Windows 95/98/NT. Nucleic acids symposium series 41:95-98.
HAUSBECK M.K., LAMOUR K.H. 2004. Phytophthora capsici on vegetable crops: Research progress and management challenges. Plant Disease 88(12):1292- 1303.
HAYWARD A.C., FEGAN N., FEGAN M., STIRLING G.R. 2010. Stenotrophomonas and Lysobacter: ubiquitous plant-associated gamma-proteobacteria of developing significance in applied microbiology. Journal of Applied Microbiology 108(3):756-770.
HERNÁNDEZ A., RIVES N., CABALLERO A., HERNÁNDEZ A.N., HEYDRICH M. 2004. Caracterización de rizobacterias asociadas al cultivo de maíz en la producción de metabolitos del tipo AIA, sideróforos y ácido salicílico. Revista Colombiana de Biotecnología 6(1):6-13.
HOWELL C.R., STIPANOVIC R.D. 1980. Suppression of Pythium ultimum induced damping-off of cotton seedlings by Pseudomonas fluorescens and its antibiotic, pyoluteorin. Phytopathology 70(8):712- 715.
HUNGRIA M., CAMPO R.J., SOUZA E.M., PEDROSA F.O. 2010. Inoculation with selected strains of Azospirillum brasilense and A. lipoferum improves yields of maize and wheat in Brazil. Plant and Soil 331(1-2):413-425.
KAYMAK H.C. 2011. Potential of PGPR in agricultural innovations, pp. 45-79. In: D.K. Maheshwari (ed). Plant growth and health promoting bacteria. Microbial Monographs vol 18.
KHALID A., ARSHAD M., ZAHIR Z.A. 2004. Screening plant growth-promoting rhizobacteria for improving growth and yield of wheat. Journal of Applied Microbiology 96(3):473-480.
KIM H.S., SANG M.K., JEUN Y.C., HWANG B.K., KIM K.D. 2008. Sequential selection and efficacy of antagonistic rhizobacteria for controlling Phytophthora blight of pepper. Crop Protection 27(3-5):436-443.
KIM K.D., NEMEC S., MUSSON G. 1997. Control of Phytophthora root and crown rot of bell pepper with composts and soil amendments in the greenhouse. Applied. Soil Ecology 5(2):169-179.
KIM S.G., KHAN Z., JEON Y.H., KIM Y.H. 2009. Inhibitory effect of Paenibacillus polymyxa GBR- 462 on Phytophthora capsici causing Phytophthora blight in chili pepper. Journal of Phytopathology 157(6):329-337.
KLOEPPER J.W., RYU C.M., ZHANG S. 2004. Induced systemic resistance and promotion of plant growth by Bacillus spp. Phytopathology 94(11):1259-1266.
KO H.S., JIN R.D., KRISHNAN H.B., LEE S.B., KIM K.Y. 2009. Biocontrol ability of Lysobacter antibioticus HS124 against Phytophthora blight is mediated by the production of 4-Hydroxyphenylacetic acid and several lytic enzymes. Current Microbiology 59(6):608-615.
LEE E.T., LIM S.K., NAM D.H., KHANG Y.H., KIM S.D. 2003. Pyoverdin 2112 of Pseudomonas fluorescens 2112 inhibits Phytophthora capsici, a red-pepper blight-causing fungus. Journal of Microbiology and Biotechnology 13(3):415-421.
MEHNAZ S., LAZAROVITS G. 2006. Inoculation effects of Pseudomonas putida, Gluconacetobacter azotocaptans, and Azospirillum lipoferum on corn plant growth under greenhouse conditions. Microbial Ecology 51(3):326-335.
MENDOZA C., PINTO B. 1985. Principios de fitopatología y enfermedades causadas por hongos. Universidad Autónoma Chapingo, México. 311 p.
OH B.T., HUR H., LEE K.J., SHANTHI K., SOH B.Y., LEE W.J., MYUNG H., KAMALA S. 2011. Suppression of Phytophthora blight on pepper (Capsicum annuum L.) by bacilli isolated from brackish environment.Biocontrol Science and Technology 21(11):1297-1311.
OLIVEIRA R.G.B., DROZDOWICZ A. 1987. Inhibition of producing strains of Azospirillum lipoferum by their own bacteriocin. Zentralblatt für Mikrobiologie 142(5):387-391.
PARK J.W., BALARAJU K., KIM J.W., LEE S.W., PARK. K. 2013. Systemic resistance and growth promotion of chili pepper induced by an antibiotic producing Bacillus vallismortis strain BS07. Biological Control 65(2):246-257.
PAUL D., SARMA Y.R. 2006. Antagonistic effects of metabolites of Pseudomonas fluorescens strains on the different growth phases of Phytophthora capsici, foot rot pathogen of black pepper (Piper nigrum L.). Archives of Phytopathology and Plant Protection 39(2):113-118.
PERVEEN R., FANI I., CHOHAN S., HAIDER S., NAZ S. 2010. Effect of nitrogen and carbon from different organic suplements on pathogenic potencial of Phytophthora capsici in the collar rot of chillies. European Journal of Scientific Research 43(1):107- 112.
PODILE A.R., KISHORE G.K. 2006. Plant growthpromoting rhizobacteria, pp.195-230. In: S.S. Gnanamanickam (ed.). Plant-Associate bacterias. Springer Netherlands.
RAJKUMAR M., LEE W.H., LEE K.J. 2005. Screening of bacterial antagonists for biological control of Phytophthora blight of pepper. Journal of Basic Microbiology 45(1):55-63.
REYES I., ALVAREZ L., EL-AYOUBI H., VALERY A. 2008. Selección y evaluación de Rizobacterias Promotoras del Crecimiento en pimentón y maíz. Bioagro 20(1):37-48.
SANG M.K., CHUN S., KIM K.D. 2008. Biological control of Phytophthora blight of pepper by antagonistic rhizobacteria selected from a sequential screening procedure. Biological Control 46(3):424-433.
SANG M.K., KIM K.D. 2012. Plant growth-promoting rhizobacteria suppressive to Phytophthora blight affect microbial activities and communities in the rhizosphere of pepper (Capsicum annuum L.) in the field. Applied Soil Ecology 62:88-97.
SCHMIDT C.S., ALAVI M., CARDINALE M., MÜLLER H., BERG G. 2012. Stenotrophomonas rhizophila DSM14405T promotes plant growth probably by altering fungal communities in the rhizosphere. Biology and Fertility of Soils 48(8):947-960.
SELVAKUMAR G., JOSHI P., NAZIM S., MISHRA P.K., BISHT J.K., GUPTA H. S. 2009. Phosphate solubilization and growth promotion by Pseudomonas fragi CS11RH1 (MTCC 8984), a psychrotolerant bacterium isolated from a high altitude Himalayan rhizosphere. Biologia 64(2):239-245.
SHAH S., KARKHANIS V., DESAI A. 1992. Isolation and characterization of siderophore, with antimicrobial activity from Azospirillum lipoferum M. Current Microbiology 25(6):347-351.
SHANAHAN P., O´SULLIVAN D.J., SIMPSON P., GLENNON J.D., O´GARA F. 1992. Isolation of 2, 4-diacetylphloroglucinol from a fluorescent pseudomonad and investigation of physiological parameters influencing its production. Applied and Environmental Microbiology 58(1):353-358.
SHARMA A., PATHAK A., SAHGAL M., MEYER J.M., WRAY V., JOHRI B.N. 2007. Molecular characterization of plant growth promoting rhizobacteria that enhance peroxidase and phenylalanine ammonia-lyase activities in chile (Capsicum annuum L.) and tomato (Lycopersicon esculentum Mill.). Archives of Microbiology 188(5):483-494.
SILVAR C., DUNCAN J.M., COOKE D.E.L., WILLIAMS N.A., DÍAZ J., MERINO F. 2005. Development of specific PCR primers for identification and detection of Phytophthora capsici Leon. European Journal of Plant Pathology 112(1):43-52.
SINGH J.S., SINGH D.P. 2013. Plant Growth Promoting Rhizobacteria (PGPR): Microbes in Sustainable Agriculture, pp. 361-385. In: A. Malik, E. Grohmann and M. Alves (eds.). Management of Microbial Resources in the Environment. Springer Netherlands.
SOLÍS V.C. 1989. Índice de enfermedades de los cultivos agrícolas de Costa Rica. Ministerio de Agricultura y Ganadería, San José. 112 p.
TAMURA K., STECHER G., PETERSON D., FILIPSKI A., KUMAR S. 2013. MEGA6: Molecular Evolutionary Genetics Analysis Version 6.0. Molecular Biology and Evolution 30(12):2725-2729.
THOMPSON J.D., GIBSON T.J., PLEWNIAK F., JEANMOUGIN F., HIGGINS D.G. 1997. The Clustal_X windows interface: flexible strategies for multiple sequence alignment aided by quality analysis tools. Nucleic Acids Research 25(24):4876- 4882.
TUSET J.J., COTS F., HINAREJOS C., MIRA J.L. 2001. Suspensión de zoosporas de Phytophthora cinnamomi que causan la “seca” en cinco especies de Quercus mediterráneos. Boletín de Sanidad Vegetal Plagas 27(1):103-115.
URIBE L., CASTRO L., ARAUZ L.F., HENRÍQUEZ C., BLANCO M. 2014. Pudrición basal causada por Phytophthora capcisi en plantas de chile tratadas con vermicompost. Agronomía Mesoamericana 25(2):243-253.
VERESOGLOU S.D., MENEXES G. 2010. Impact of inoculation with Azospirillum spp., on growth properties and seed yield of wheat: A meta-analysis of studies in the ISI Web of Science from 1981 to 2008. Plant and Soil 337(1-2):469-480.
YANG M.M., XU L.P., XUE Q.Y., YANG J.H., XU Q., LIU H.X., GUO J.H. 2012. Screening potential bacterial biocontrol agents towards Phytophthora capsici in pepper. European Journal of Plant Pathology 134(4):811-820.
ZHANG S., WHITE T.L., MARTINEZ M.C., McINROY J.A., KLOEPPER J.W., KLASSEN W. 2010. Evaluation of plant growth-promoting rhizobacteria for control of Phytophthora blight on squash under greenhouse conditions. Biological Control 53(1):129-135.