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Abstract: Fusarium head blight (FHB) caused by Fusarium graminearum is a devastating disease that results in extensive yield losses to wheat and barley. A green fluorescent protein (GFP) expressing plasmid pRP22-GFP was constructed for monitoring the colonization of two biocontrol agents, Brevibacillus brevis ZJY-1 and Bacillus subtilis ZJY-116, on the spikes of barley and their effect on suppression of FHB. survival and colonization of the Brevibacillus brevis ZJY-1 and Bacillus subtilis ZJY-116 strains on spikes of barley were observed by tracking the bacterial transformants with GFP expression. Our field study revealed that plasmid pRP22-GFP was stably maintained in the bacterial strains without selective pressure. The retrieved GFP-tagged strains showed that the bacterial population fluctuation accorded with that of the rain events. Furthermore, both biocontrol strains gave significant protection against FHB on spikes of barley in fields. The greater suppression of barley FHB disease was resulted from the treatment of barley spikes with biocontrol agents before inoculation with F. graminearum.

[1] Araujo, M.A.V., Mendonca-Hagler, L.C., Hagle, A.N., 1994. Survival of genetically modified Pseudomonas fluorescences introduced into subtropical soil microcosms. FEMS Microbiol. Ecol., 13:205-216.

[2] Bennett, A.J., Leifert, C., Whipps, J.M., 2003. Survival of the biological agents Coniothyrium minitans and Bacillus subtilis MBI600 introduced into pasteurized, sterilised and non-sterile soils. Soil Biol. Biochem., 35:1565-1573.

[3] Boelens, J., Woestyne, M.V., Verstraele, W., 1994. Ecological importance of motility for the plant growth-promoting rhizopseudomonas strain ANP15. Soil Biol. Biochem., 26:269-277.

[4] Capettini, F., Rasmusson, D.C., Dill-Macky, R., Schiefelbein, E., Elakkad, A., 2003. Inheritance of resistance to fusarium head blight in four populations of barley. Crop Sci., 43:1960-1966.

[5] Chang, S., Cohen, S.N., 1979. High frequency transformation of Bacillus subtilis protoplasts by plasmid DNA. Molec. Gen. Genet., 168:111-115.

[6] Chao, W.L., Nelson, E.B., Harman, G.E., Hoch, H.C., 1986. Colonization of the rhizosphere by biological control agents applied to seeds. Phytopathology, 76:60-65.

[7] Cho, S.J., Lee, S.K., Cha, B.J., Kim, Y.H., Shin, K.S., 2003. Detection and characterization of the Gloeosporium gloeosporioides growth inhibitory compound iturin A from Bacillus subtilis strain KS03. FEMS Microbiol. Lett., 223:47-51.

[8] Douka, E., Christogianni, A., Koukkou, A.I., Afendra, A.S., Drainas, C., 2001. Use of green fluorescent protein gene as a reporter in Zymomonas mobilis and Halomonas elongata. FEMS Microbiol. Lett., 201:221-227.

[9] Edwards, S.G., Seddon, B., 1992. Bacillus brevis as a Biocontrol Agent against Botrytis cinerea on Protected Chinese Cabbage. In: Recent Advances in Botrytis Research. Pudoc Scientific Publications, The Netherlands, p.267-271.

[10] Edwards, S.G., Seddon, B., 2001. Mode of antagonism of Brevibacillus brevis against Botrytis cinerea in vitro. J. Appl. Microbiol., 91:652-659.

[11] Elizabeth, A.B., Emmert, H.J., 1999. Biocontrol of plant disease: A (Gram-)positive perspective. FEMS Microbiol. Lett., 171:1-9.

[12] Handelsman, J., Stabb, E., 1996. Biocontrol of soilborne plant pathogen. Plant Cell, 8:1855-1869.

[13] Ingalls, J.R., 1996. Influence of deoxynivalenol on feed consumption by dairy cows. Anim. Feed Sci., 60:297-300.

[14] Kim, P., Chung, K.C., 2004. Production of an antifungal protein for control of Colletotrichum lagenarium by Bacillus amyloliquefaciens MET0908. FEMS Microbiol. Lett., 234:177-183.

[15] Klement, Z., Rudolph, K., Sands, D.C., 1990. Methods in Phytobacteriology. Academiai Kiado, Budapest.

[16] Kragelund, L., Christoffersen, B., Nybroe, O., de Bruijn, F.J., 1995. Isolation of lux report gene fusions in Pseudomonas fluorescences DF57 inducible by nitrogen or phosphorus starvation. FEMS Microbiol. Ecol., 17:95-106.

[17] Lewis, P.J., Marston, A.L., 1999. GFP vectors for controlled expression and dual labeling of protein fusions in Bacillus subtilis. Gene, 227:101-109.

[18] Mao, W., Lewis, J.A., Hebbar, P.K., Lumsden, R.D., 1997. Seed treatment with a fungal or a bacterial antagonist for reducing corn damping-off caused by species of Pythium and Fusarium. Plant Dis., 81:450-454.

[19] Mavrodi, O.V., Gardener, B.B.M., Mavrodi, D.V., Bonsall, R.F., Weller, D.M., Thomashow, L.S., 2000. Genetic diversity of phlD from 2,4-diacetylphloroglucinol-producing Fluorescent pseudomonas spp. Phytopathology, 91:35-43.

[20] Miller, J.H., 1972. Determination of Viable Cell Count: Bacterial Growth Curves. In: Experiments in Molecular Genetics. 2nd Ed., Cold Spring Harbour Laboratory, New York, USA, p.31-36.

[21] Nautiyal, C.S., Johri, J.K., Singh, H.B., 2002. Survival of the rhizosphere-competent biocontrol strains Pseudomonas fluorescences NBR12650 in the soil and phytosphere. Can. Microbiol., 48(7):588-601.

[22] O’Callaghan, M., Gerard, E.M., Johnson, V.W., 2001. Effect of soil moisture and temperature on survival of microbial control agents. New Zeal. Plant Protect., 54:128-135.

[23] Olsson, J., Bőrjesson, T., Lundstedt, T., Schnürer, J., 2002. Detection and quantification of ochratoxin A and deoxynivalenol in barley grains by GC-MS and electronic nose. Int. J. Food Microbiol., 72:203-214.

[24] Onji, Y., Aoki, Y., Tani, N., Umebayashi, K., Kitada, Y., Dohi, Y., 1998. Direct analysis of several Fusarium mycotoxins in cereals by capillary gas chromatography-mass spectrometry. J. Chromatogr. A., 815:59-65.

[25] Placinta, C.M., D’Mello, J.P.F., Macdonald, A.M.C., 1999. A view of worldwide contamination of cereal grains and animal feed with Fusarium mycotoxins. Anim. Feed Sci. Tech., 78:21-37.

[26] Ramos, H.J.O., Roncato-Maccari, L.D.B., Souza, E.M., Soares-Ramos, J.R.L., Hungria, M., Pedrosa, F.O., 2002. Monitoring Azospirillum-wheat interaction using the gfp and gusA genes constitutively expressed from a new broad-host range vector. J. Biotechnology, 97:243-252.

[27] Saito, Y., Otani, S., Otani, S., 1970. Biosynthesis of gramicidin S. Adv. Enzymol., 33:337-380.

[28] Scott, K.P., Mercer, D.K., Glover, L.A., Flint, H.J., 1998. The green fluorescent protein as a visible marker for lactic acid bacteria in complex ecosystems. FEMS Microbiol. Ecol., 26:219-230.

[29] Somasegaran, P., Hoben, H.J., 1994. Handbook for Rhizobia: Method in Legume-Rhizobium Technology. Springer Publications, New York, USA.

[30] Thomashow, L.S., Weller, D.M., 1988. Role of a phenazine antibiotic from Pseudomonas fluorescens in biological control of Gaeumannomyces graminis var. tritici. J. Bacteriology, 170:3499-3508.

[31] Urrea, C.A., Horsley, R.D., Steffenson, B.J., Schwarz, P.B., 2002. Heritability of fusarium head blight resistance and deoxynivalenol accumulation from barley accession CIho 4196. Crop Sci., 42:1404-1408.

[32] Wilson, K.J., 1995. Molecular technology for the study of rhizobial ecology in the field. Soil Biol. Biochem., 27:501-514.

[33] Xi, C.W., Lambrecht, M., Vanderleyden, J., Michiels, J., 1999. Bi-functional gfp-and gusA-containing mini-Tn5 transposon derivatives for combined gene expression and bacterial localization studies. J. Microbiol. Meth., 35:85-92.

[34] Yu, G.Y., Sinclair, J.B., Hartman, G.L., Bertagnolli, B.L., 2002. Production of iturin A by Bacillus amyloliquefaciens suppressing Rhizoctoria solani. Soil Biol. Biochem., 34:955-963.