Endophytic bacteria of desert cactus (Euphorbia trigonas Mill) confer drought tolerance and induce growth promotion in tomato (Solanum lycopersicum L.).

Endophytic bacteria isolated from cactus were characterized and assessed for their capability to induce drought tolerance and growth promotion in tomato. A total of 191-bacteria representing 13-genera and 18-species were isolated from wild cactus, Euphorbia trigonas. Bacillus (58), Lysinibacillus (36), Enterobacter (29), Stenotrophomonas (18), Lelliottia (12) and Pseudomonas (12) were the most represented genera. 16S rDNA sequence (>1400-bp) comparison placed the bacterial isolates with Bacillus xiamenensis; Bacillus megaterium; Bacillus cereus; Bacillus amyloliquefaciens; Bacillus velezensis; Brevibacillus brevis; Lysinibacillus fusiformis; Enterobacter cloacae; Lelliottia nimipressuralis; Proteus penneri; Sphingobacterium multivorum; Klebsiella pneumoniae; Pseudomonas putida; Pseudomonas aeruginosa; Stenotrophomonas maltophilia; Citrobacter freundii; Chryseobacterium indologenes and Paracoccus sp. Bacillus xiamenensis was identified for the first time as plant endophyte. Upon bacterization, the endophytes triggered germination and growth promotion in tomato as indicated by 118 % and 52 % more root-biomass under drought-free and drought-induced conditions, respectively. Bacillus amyloliquefaciens CBa_RA37 and B. megaterium RR10 displayed broad spectrum endophytism in tomato. Bacterization of tomato with cactus endophyte showed altered oxidative status, stomatal and photosystem II functioning, internal leaf temperature and relative water content suggestive of physiological de-stressing from moisture stress. Activity of oxidative stress enzymes such as guaiacol peroxidase and catalase was also indicative of endophyte assisted de-stressing of tomato. Re-irrigation on 20-days of drought infliction showed 86.9% recovery of B. amyloliquefaciens CBa_RA37 primed tomato when non-primed plantlets succumbed. The cactus endophytic bacterial strain B. amyloliquefaciens CBa_RA37 showed promise for low-cost, efficient and environmentally friendly bio-inoculant technology to mitigate drought in arid zones of Asian and African continents.

Eke P ，Kumar A ，Sahu KP ，Wakam LN ，Sheoran N ，Ashajyothi M ，Patel A ，Fekam FB ... -  《-》

Endophytes from Gnetum gnemon L. can protect seedlings against the infection of phytopathogenic bacterium Ralstonia solanacearum as well as promote plant growth in tomato.

Endophytes are beneficial plant microbes which help the plants by producing various plant growth promoting substances and also by acting as biocontrol agents against various plant pathogens. In the present study, evaluation of endophytic bacteria isolated from Gnetum gnemon, an ethnomedicinal plant was carried out for their plant growth promoting (PGP) activity and antagonistic potential against bacterial wilt pathogen Ralstonia solanacearum. Initially a total of 40 endophytic bacteria were isolated which were clustered into 13 groups based on RFLP and BOX-PCR fingerprinting. These 13 representative isolates belonged to different genera of Bacillus (9), Solibacillus (1), Staphylococcus (2) and Caballeronia (1). Among them, six isolates were positive for production of IAA, the value of which ranged from 11.16 to 27.63 μg mL-1. Phosphate solubilisation in the range of 106.4 to -212.7 μg mL-1 was shown by three isolates. Eight isolates producedammonia, the value of which ranged from 1.3 to 6.1 μmol mL-1. All tested isolates were positive for siderophore production. For extracellular enzyme production, 7 isolates were positive for protease, 8 for cellulase and 10 for amylase production. The isolates were also tested for their antagonistic activity against R. solanacearum in vitro and in planta assay using tomato seedlings. Staphylococcus warneri GL1 showed the highest biocontrol efficacy of 77.67 % followed by Bacillus velezensis GL3 i.e. 70.1 %. R. solanacearum antagonistic isolates were analysed for the presence of antimicrobial peptide biosynthesis genes bmyB, srfAA, fenD and ituC. All the antagonistic isolates showed the presence of all four genes, except the isolate Bacillus velezensis GMC2, where the gene for fengycin synthetase (fenD) was absent. Based on in vitro PGP traits, three isolates Bacillus velezensis GL3, Bacillus atrophaeus GMC1 and Bacillus megaterium GS2 were selected, these three endophytic bacteria individually and their consortia were tested for in planta PGP activities in tomato plants. Application of Bacillus velezensis GL3 alone and consortia of three isolates showed significant improvement in growth parameters such as shoot length, fresh weight and dry weight in a pot experiment. Colonization of endosphere of treated tomato seedlings by the endophytic isolate Bacillus velezensis GL3 was confirmed by visualization of colony morphology and BOX-PCR fingerprinting. Our study highlights the potential of endophytes associated with unexplored plants like G. gnemon for development of bioformulation aimed at enhancing plant growth and bacterial wilt disease control.

Agarwal H ，Dowarah B ，Baruah PM ，Bordoloi KS ，Krishnatreya DB ，Agarwala N ... -  《-》

Diversity, antimicrobial and antioxidant activities of culturable bacterial endophyte communities in Aloe vera.

Twenty-nine culturable bacterial endophytes were isolated from surface-sterilized tissues (root, stem and leaf) of Aloe vera and molecularly characterized to 13 genera: Pseudomonas, Bacillus, Enterobacter, Pantoea, Chryseobacterium, Sphingobacterium, Aeromonas, Providencia, Cedecea, Klebsiella, Cronobacter, Macrococcus and Shigella. The dominant genera include Bacillus (20.7%), Pseudomonas (20.7%) and Enterobacter (13.8%). The crude and ethyl acetate fractions of the metabolites of six isolates, species of Pseudomonas, Bacillus, Chryseobacterium and Shigella, have broad spectral antimicrobial activities against pathogenic Pseudomonas aeruginosa, Staphylococcus aureus, Bacillus cereus, Salmonella Typhimurium, Proteus vulgaris, Klebsiella pneumoniae, Escherichia coli, Streptococcus pyogenes and Candida albicans, with inhibition zones ranging from 6.0 ± 0.57 to 16.6 ± 0.57 mm. In addition, 80% of the bacterial endophytes produced 1,1-diphenyl-2-picrylhydrazyl (DPPH) with scavenging properties of over 75% when their crude metabolites were compared with ascorbic acid (92%). In conclusion, this study revealed for the first time the endophytic bacteria communities from A. vera (Pseudomonas hibiscicola, Macrococcus caseolyticus, Enterobacter ludwigii, Bacillus anthracis) that produce bioactive compounds with high DPPH scavenging properties (75-88%) and (Bacillus tequilensis, Pseudomonas entomophila, Chryseobacterium indologenes, Bacillus aerophilus) that produce bioactive compounds with antimicrobial activities against bacterial pathogens. Hence, we suggest further investigation and characterization of their bioactive compounds.

Akinsanya MA ，Goh JK ，Lim SP ，Ting AS ... -  《-》

Phytohormone production endowed with antagonistic potential and plant growth promoting abilities of culturable endophytic bacteria isolated from Clerodendrum colebrookianum Walp.

In this study, culturable endophytic bacterial isolates obtained from an ethnomedicinal plant Clerodendrum colebrookianum Walp., were assessed for their diversity, in vitro screening for their plant growth promoting (PGP) activities and to use them as inoculant for in vivo PGP activities with biocontrol potential. Totally, 73 isolates were recovered from different tissues of C. colebrookianum were identified by 16S rRNA gene sequencing and phylogenetically analyzed by using BOX-PCR fingerprinting. Out of 73 isolates, 52 exhibited varying extents of antagonistic potential were selected for screening for various PGP traits. Concerning the PGP activities, the percentage of isolates positive for P-solubilisation, indolic compounds production, siderophore and ammonia production were 84.6, 92.3, 78.8 and 98.0 respectively. All isolates were positive for the production of hydrocyanic acid (HCN) and 86.5%, 84.6% and 90.3% of isolates showed significant cellulase, amylase and protease production respectively. Further, the top 10 bacterial isolates based on a bonitur scale with multiple PGP activities were screened for root surface colonization and biofilm formation ability. Out of selected 10 isolates, 9 showed significant potential for root surface colonization on tomato roots. Isolate BPSAC6 identified as Bacillus sp. was most efficient in biofilm formation as assessed with respect to the intensity of crystal violet, which further showed their potential to withstand various biotic and abiotic stresses. Furthermore, Bacillus sp. strain BPSAC6 showed a significant increase in shoot and root height as well as fresh weight after 45 and 60 d of inoculation with tomato seedlings. Additionally, biosynthetic potential of antagonistic isolate was detection by using PKSI, PKSII and NRPS biosynthetic genes. Two isolates Pseudomonas psychrotolerans and Labrys wisconsinensis were reported first time as an endophyte. At last, first time an endophytic bacterial strain Bacillus sp. BPSAC6 was reported to produce altogether three phytohormones (IAA, Kinetin and 6-Benzyladenine). This study is the first report that bacteria isolated from C. colebrookianum has biocontrol as well as PGP abilities endowed with phytohormones production and can be used for the preparation of bioinoculant for plant growth promotion.

Passari AK ，Mishra VK ，Leo VV ，Gupta VK ，Singh BP ... -  《-》

The communities of tomato (Solanum lycopersicum L.) leaf endophytic bacteria, analyzed by 16S-ribosomal RNA gene pyrosequencing.

Endophytic bacterial communities of tomato leaves were analyzed by 16S-rRNA gene pyrosequencing and compared to rhizosphere communities. Leaf endophytes mainly comprised five phyla, among which Proteobacteria was the most represented (90%), followed by Actinobacteria (1,5%), Planctomycetes (1,4%), Verrucomicrobia (1,1%), and Acidobacteria (0,5%). Gammaproteobacteria was the most abundant class of Proteobacteria (84%), while Alphaproteobacteria and Betaproteobacteria represented 12% and 4% of this phylum, respectively. Rarefaction curves for endophytic bacteria saturated at 80 OTUs, indicating a lower diversity as compared to rhizosphere samples (> 1700 OTUs). Hierarchical clustering also revealed that leaf endophytic communities strongly differed from rhizospheric ones. Some OTUs assigned to Bacillus, Stenotrophomonas, and Acinetobacter, as well as some unclassified Enterobacteriaceae were specific for the endophytic community, probably representing bacteria specialized in colonizing this niche. On the other hand, some OTUs detected in the leaf endophytic community were also present in the rhizosphere, probably representing soil bacteria that endophytically colonize leaves. As a whole, this study describes the composition of the endophytic bacterial communities of tomato leaves, identifying a variety of genera that could exert multiple effects on growth and health of tomato plants.

Romero FM ，Marina M ，Pieckenstain FL 《-》