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Journal of Agricultural Biotechnology and Sustainable Development Vol. 2(8) pp. xxx-xxx, April, 2010Available online http://www.academicjournals.org/JABSD

2010 Academic Journals

Full Length Research Paper

Biofertilization of banana (Musaspp. L.) with free-livingN2 fixing bacteria and their effect on mycorrhization and

the nematode Radopholus similis

Adriano-Anaya ML1, Gutirrez-Miceli FA2, Dendooven L3, Salvador-Figueroa M1*

1Centro de Biociencias. Universidad Autnoma de Chiapas. Carretera a Puerto Madero Km 2.0 Tapachula, 30700,

Chiapas, Mxico.2Departamento de Ingeniera Qumica y Bioqumica. Instituto Tecnolgico de Tuxtla-Gutirrez, Tuxtla-Gutirrez,

Chiapas, Mxico.3Laboratory of Soil Ecology, GIB, Dept. Biotechnology and Bioengineering, Cinvestav, Mxico.

November 24, 2010

Banana plants (Musaspp. L.) cv 'Great dwarf' were amended with free-living N2 fixing bacteria (FLNFB)and arbuscular mycorrhizal fungi (AMF) and the presence of the burrowing nematode Radopholussimiliswas monitored in the field. Five treatments were applied by inoculating banana roots with fourstrains of FLNFB, that is C1, C2, C3 and C4 isolated from the rhizoplane of the same banana cultivar, orby keeping them uninoculated. The largest number of nematodes was found in the untreated roots andthe lowest in the roots inoculated with C4. The largest percent of mycorrhizal colonization was foundwhen banana roots were inoculated with C1 and the lowest in roots that were not inoculated. Thenumber of R. similis decreased with increased colonization with AMF. The largest percentage ofnecrosis of the banana roots was found when banana roots were not inoculated and the lowest in rootsthat were inoculated with C4. The necrosis of the roots increased with increased number of R. similis,but decreased with increased colonization with AMF. It was found that inoculating the banana cv 'Greatdwarf' with FLNFB increased the root colonization with AML, and reduced the number of nematodesand root damage.

Key words: Arbuscular mycorrhizal fungi, burrowing or banana-root nematode, free-living N2 fixing bacteria,plant development.

INTRODUCTION

Plant development is closely related to the interactionwith microbiota in the phylosphere and rhizosphere(Green et al., 2006). For instance, 80 and 92% of

surveyed plant species and families, respectively, weremycorrhizal with arbuscular mycorrhizal fungi (AMF)being dominant (Wang and Qiu, 2006). They suggestedthat the origin of arbuscular mycorrhiza coincides with theorigin of land plants. In the rhizosphere, AMF andpathogens need to be in equilibrium, so that a normal

*Corresponding author: E-mail: [email protected], Tel. +52(962) 6427972.

plant development can occur (Reyes and Valery, 2007)The increase in pathogens populations leads to diseases(Whipps, 2001) thereby hampering plant growth (Siddiqu

and Akhtar, 2008). The interaction in the rhizospherebetween plant and microorganisms has been studiedintensively (Mathesius, 2009) and the importance ocertain root exudates in the relationship between plantsand microorganisms, beneficial and pathogenic, hasbeen demonstrated (Rojas-Andrade et al., 2003)Radopholus similis, the burrowing or banana-roonematode, is one of the most important root parasites ofbanana plants (Musa spp. L.). Although effectivechemical nematicides exist, but their high cost andtoxicity limit their use. Nematode management for


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Figure 1. The number of nematodes Radopholus similis(per 100 g roots), b) the percentage of arbuscular

mycorrhizal fungi colonization (%) and c) necrosis (%) of the roots of the banana plant (Musaspp. L.) cv'Great dwarf' inoculated with four different free-living N2 fixing bacteria (C1 (), C2 (), C3 (), C4 ()) andleft untreated () grown in the field.

roots. The number of nematodes found on the rootsremained low until day 128 and started to increasethereafter (Figure 1a). At day 240, the number ofnematodes found in the roots was significantly differentbetween the treatments (P


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Figure 2. The correlation a) between the number of nematodes Radopholus similis(per 100 g roots) andthe percentage of arbuscular mycorrhizal fungi colonization (%), b) the number of nematodes Radopholussimilis(per 100 g roots) and necrosis (%) of the roots c) necrosis (%) of the roots and the percentage ofarbuscular mycorrhizal fungi colonization (%) of the roots of the banana plant (Musa spp. L.) cv 'Greatdwarf' inoculated with four different free-living N2 fixing bacteria.

cereals, AMF root colonization is related to thephenological stage of the plant (Gavito and Varela, 1993;Majic et al., 2008). Colonization increased during thevegetative phase and decreased during the reproductivephase. Photosynthetic products are transported to theroots where they are used by the microorganisms in therhizosphere. The microorganisms proliferate and theirmetabolites and/or released nutrients are taken up by theplants. When the plant matures and the reproductivephase starts, photosynthetic products are used forflowering and seed development. The plant metabolites

released in the rhizosphere decreases, microbial growthslows and the AMF loosen their association with corticacells.

The level of colonization with AMF found in thisexperiment for banana plants not inoculated were lessthan those reported. Adriano (2001) found a 58 to 88%colonization with Glomus mosseae for the banana cv'Great dwarf' during the reproductive phase. Jaizme-Vegaand Pinochet (1997) found a 47% colonization of roots othe same banana cultivar with G. mosseae, 34% with Gaggregatumand 33% with G. intraradices.


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Population of Radopholus similis

Jaizme-Vega et al. (1997) reported that the banana cv'Great dwarf' colonized by the AMF G. mosseae wereless susceptible to infections with the root-knot nematodeMeloidogyne incognitafavouring plant growth. Contrarily,Jaizme-Vega and Rodriguez-Romero (2004) reportedthat the same banana plants inoculated with G. mosseae,G. aggregatumand G. intraradiceshad the same numberof Pratylenchus goodeyi and M. incognita than theuntreated plants. However, Elsen et al. (2008) found thatAMF have the ability to induce systemic resistanceagainst plant parasitic nematodes R. similis andPratylenchus coffeae in the root system. AMF reducedboth nematode species by more than 50%, even whenthe AMF and the plant parasitic nematodes were spatiallyseparated. Similar results were obtained with maize (ZeamaysL.), olive (Olea europaeaL.) (Castillo et al., 2006)

and kiwi (Actinidia deliciosa) (Verdejo et al., 1990). Themycorrhiza had neither effect on the number ofpathogenic nematodes nor was the number of AMLaffected by the pathogens.

Root damage

Jaisme-Vega et al. (1997) reported a 36 to 64% reductionin root galling caused by the root-knot nematode M.incognitawhen micropropagated banana plants cv 'Greatdwarf' were inoculated with G. mosseae reaching 58 to88% of AM root colonization. Jaizme-Vega and

Rodriguez-Romero (2004) found that banana plants cv'Great dwarf' inoculated with G. mosseae, G. aggregatumand G. intraradices, and infected with the nematode P.goodeyi, had 4, 15 and 13% lesions, respectively, whilethe plants that were not inoculated had 28%. Thecolonization with G. mosseae was 47%, with G.aggregatum 34% and with G. intraradices 33%. Nodifferences, however, in nematode populations werefound. Elsen et al. (2008) found that the number oflesions in the roots of banana cv Williams amended with1850 mycorrhizal spores and infected with 5000 eggsand juvenile M. javanica one month later, were 29%lower with G. mosseae, 25% with G.macrosporum and16% with G. caledonium compared to plants notinoculated with mycorrhizae. They suggested aninhibitory effect of the micorrhiza on the infection withnematodes through changes in the nutrition of the plant,biochemical changes in the plant tissue, e.g. an increasein aminoacids, peroxidases, quitinases, phytoalexins,increases in lignin, stress reduction, changes in themicrobial populations in the roots and a more dense rootsystem. Vaast et al. (1998) found a decrease in lesionsby P. coffeae in the roots of coffee (Coffea arabica L.)when inoculated with the mycorrhizae Acaulosporamelleaor G. clarum. Castillo et al. (2006) found similar

results when studying the effect of mycorrhiza on thepathogenicity of M. incognita and P. vulnus on roots oolive tree. Castillo et al. (2006) suggested that thereduction in pathogenicity was not related to a reductionin infection rate, but was related to a higher tolerancetowards the nematode.

Our findings conclude that inoculating roots of thebanana plant cv 'Great dwarf' with free-living N2 fixingbacteria isolated from roots of the similar plant resulted ina higher colonization of the roots with AMF, a lowernumber of pathogenic nematodes and healthier roots.

ACKNOWLEDGEMENTS

We thank FOMIX CONACyT-Estado de Chiapas forfunding the research (Grant CHIS-2002-C01-4332).

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