aerial parts are used in the treatment of kidney stones , hypertension , fever and cough ( http://www.forestry.gov.my/ ) . the earlier studies have shown that endophytic microorganisms isolated from medicinal plants produce the same metabolites as their hosts . therefore , there is a great potential in exploring endophytes as a source of therapeutic natural products . however , despite several traditional medicinal applications of nepenthes spp . the objective of this study was to isolate and to identify the endophytic bacteria ( eb ) from nepenthes spp . namely , nepenthes ampullaria , n. gracilis , n. macfarlanei , n. mirabilis , n. rafflesiana and n. sanguinea were collected from frim , selangor , malaysia . however , leaves and twigs from one to three individual plants of nepenthes spp . , namely , n. alba , n. albomarginata , n. gracillima and n. sanguinea were collected from the gunung jerai ( gj ) , gurun , kedah , malaysia . leaves with petioles were thoroughly washed under running tap - water and the surface - sterilization of plant material samples was carried out as reported elsewhere . the stem pieces were soaked in 70% ethanol and flamed to make their surface sterile . aseptically , the leaf and stem tissue pieces were inoculated in the petri plates containing luria - bertani ( lb ) agar medium . the plates were incubated in an incubator at 37c ( 3c ) for 18 - 20 h in the dark . the isolation , cultivation of endophytes , amplification of 16s rdna , sequencing of 16s rdna , identification of endophytes and rooted phylogenetic tree construction was carried out as reported by bhore et al . incubation of the inoculated leaf discs and stems pieces on lb agar medium enabled cultivable eb to grow , and the colonies of grown eb were visible on the margins of the leaf and stem tissues . all 96 isolates were identified based on 16s rdna sequence blast ( megablast ) hits analysis . the annotated 16s rrna gene fragment ( 16s rdna ) nucleotide sequences of all isolates have been submitted to the international dna database ( genbank / ddbj / embl ) under accession numbers : jf819686-jf819713 and jf938974-jf939041 . were from the bacilli ( 59.4% ) class , followed by gammaproteobacteria ( 35.4% ) and betaproteobacteria ( 5.2% ) [ table 1 ] . the genera of endophytic bacterial isolates ( ebis ) isolated from 9 nepenthes plant species the 16s rdna multiple sequence alignment output from clustalw was used in the construction of a rooted dendrogram . rooted dendrogram showing clustering of 44 diverse types of endophytic bacteria ( eb ) isolated from nine nepenthes spp . the accession number of 16s rdna ( rrna gene ) sequence of the respective isolate is given in front of the species name in this short and snappy study , we isolated and identified 96 isolates from nine nepenthes spp . eb have been reported from several medicinal plants , for instance glycyrrhiza spp . however , to the best of our knowledge , our study is the first to illustrate diversity and types of eb in nine stated nepenthes spp . comparison between the annotated 16s rdna fragment sequences from isolates and the sequences from genbank / ddbj / embl database using the blastn program revealed the identity of the respective isolates . the rooted dendrogram clearly showed the clustering of the species from the bacilli , betaproteobacteria and gammaproteobacteria groups . endophytes are found abundantly in various plant species studied to date , and soil bacteria such as bacillus spp . , pseudomonas spp . and azospirillum spp . however , we did not find any pseudomonas spp . and azospirillum spp . in our isolates . during isolation of eb , the growth medium used might be directly affecting the number and type of endophytic microorganisms that can be isolated from the plant tissues . the tissue samples used in the isolation of eb were from a single or few plants of each nepenthes spp . the location and the conditions in which plant species are grown also determine the types of endophytes in it . we have used plant samples from nepenthes spp . that were collected from their wild habitat in gj and diverse collection available at frim . this could be the reason for the wide diversity of eb in the studied nepenthes spp . it is important to note that from 96 isolates , 22 isolates ( representing 15 species ) belonged to the enterobacteriaceae family , which contains human enteric pathogens . a number of species from the enterobacteriaceae family have been reported as endophytes , viz entrobacter cloacae and klebsiella pneumonia in maize , entrobacter asburiae in cotton , and klebsiella spp . and entrobacter cloacae in banana . of the 44 species of eb isolated from nepenthes spp . however , we have not found any published record that reported acinetobacter soli , bacillus cibi , b. horneckiae , b. indicus , b. koreensis , b. stratosphericus , citrobacter gillenii , c. youngae , kluyvera ascorbata , providencia alcalifaciens and serratia liquefaciens as endophytes . perhaps , this is the first study that reports these bacterial species as endophytes . however , the benefits derived by nepenthes spp . from these bacterial endophytes and its quantum are not clearly understood yet . from this study contains diverse types of cultivable eb , and that the majority of bacterial endophytes ( 59.4% ) were from the bacilli class . nonetheless , these research findings could serve as a foundation in further research on the therapeutic properties of nepenthes spp . in correlation with their bacterial endophytes . we hypothesize that in nepenthes spp . , these eb might be involved in producing bioactive compounds of pharmaceutical importance , and further research is required to ascertain the same .