title: Endophytic bacterial communities associated with two explant sources of Eucalyptus benthamii Maiden & Cambage.
abstract: Micropropagation has been applied in the recovery and rejuvenation of adult trees, which is achieved by various subcultures in the multiplication phase. This strategy has brought questions about the endophytic microbiota associated with these plants along its manipulation. Therefore, the aim of this study was to evaluate the composition of the endophytic bacterial communities associated with two explants sources [the canopy branches (CB) and the trunk base of the tree (TB)] under prolonged in vitro cultivation. In addition we analyzed the bacterial community dynamic along the subcultures in different micropropagation phases. Bacterial DNA was extracted from samples of mini-stumps (in vivo) from CB and TB and in micro-stumps produced by in vitro cultivations of these explants sources--both originated from one single matrix plant of Eucalyptus benthamii. In vitro establishment occurred in two dates and the evaluation of endophytic bacterial communities was made in vivo and in vitro samples (on 10th, 13th and 16th subcultures), when elongated shoots and roots were analyzed. Analysis was performed by PCR-DGGE based on the V6 region of ribosomal gene 16S rDNA. Bands profiles showed differences in communities between in vivo and in vitro samples, and also distinctions of communities assessed in the subcultures, elongated and rooted samples. Distinctions in the composition of endophytic bacterial communities were greater in CB micro-stumps. These results indicate a differential colonization of explants by endophytic bacteria, with predominance of common (ever-present) endophytes in TB samples and casual, here named opportunistic, in CB samples.
authors.affiliation: Biological Science Department, "Luiz de Queiroz" Superior College of Agriculture, University of São Paulo (ESALQ/USP), Pádua Dias Avenue, 11, P.O. Box 9, Piracicaba, SP, 13418-900, Brazil. esposito.polesi@gmail.com.
authors.forename: Natalia Pimentel
authors.initials: NP
authors.lastname: Esposito-Polesi
authors.affiliation: Soil Science Department, ''Luiz de Queiroz'' Superior College of Agriculture, University de São Paulo (ESALQ/USP), Pádua Dias Avenue, 11, P.O. Box 9, Piracicaba, SP, 13418-900, Brazil.
authors.forename: Pedro Avelino Maia
authors.initials: PA
authors.lastname: de Andrade
authors.affiliation: In Vitro Palm Consultoria, Estudo e Desenvolvimento Biológico Ltda, Rua Itajobi, 421, Piracicaba, SP, 13432-000, Brazil.
authors.forename: Cristina Vieira
authors.initials: CV
authors.lastname: de Almeida
authors.affiliation: Soil Science Department, ''Luiz de Queiroz'' Superior College of Agriculture, University de São Paulo (ESALQ/USP), Pádua Dias Avenue, 11, P.O. Box 9, Piracicaba, SP, 13418-900, Brazil.
authors.forename: Fernando Dini
authors.initials: FD
authors.lastname: Andreote
authors.affiliation: Biological Science Department, "Luiz de Queiroz" Superior College of Agriculture, University of São Paulo (ESALQ/USP), Pádua Dias Avenue, 11, P.O. Box 9, Piracicaba, SP, 13418-900, Brazil.
authors.forename: Marcílio
authors.initials: M
authors.lastname: de Almeida
mesh_terms: D001419:Bacteria / Q000145:classification* / Q000235:genetics; D004269:DNA, Bacterial / Q000235:genetics; D004275:DNA, Ribosomal / Q000235:genetics; D060026:Endophytes / Q000145:classification* / Q000235:genetics; D005052:Eucalyptus / Q000382:microbiology*; D066298:In Vitro Techniques; D010802:Phylogeny; D018517:Plant Roots / Q000382:microbiology; D018520:Plant Shoots / Q000382:microbiology; D012336:RNA, Ribosomal, 16S / Q000235:genetics; D017422:Sequence Analysis, DNA / Q000379:methods
chemical_list: D004269:DNA, Bacterial; D004275:DNA, Ribosomal; D012336:RNA, Ribosomal, 16S
keywords: Eucalyptus; Explants; Micropropagation; Successive subcultures