Isolation and identification of rhizospheric pseudomonads with insecticidal effects from various crops in Khuzestan Province, Iran.

Pseudomonas bacteria include a variety of species with distinct characteristics. Some species within this genus are known for their ability to stimulate plant growth. Recently, the potential of these bacteria in controlling insect pests has been documented. In this study, 58 bacterial isolates were purified from rhizospheres of wheat, broad bean and canola that were collected from different fields of Khuzestan province in south-west of Iran. With biochemical tests 19 non plant pathogenic pseudomonads strains were detected and their lethal effects on the eggs and larvae of Ephestia keuhniella as an important pest that infests stored products, were evaluated under laboratory conditions. For the bioassays, two concentrations of each strain were administered, and the 5th instar larvae and eggs of the pest were subjected to treatment. Mortality rates were recorded after 24 h. The results showed that all isolated Pseudomonad strains of this study had insecticidal effects against eggs and larvae of E. keuhniella. The strains AWI1, AWI2, AWI7, ABI12, ABI15 and ABI16 displayed the highest mortality rate (91.1 %, 86.2 %, 82.3 %, 84.2, 90.5 % and 90.5 %, respectively). Molecular identification and phylogeny tree according to 16 s rRNA sequencing clarified that AWI1, AWI2 belong to P. plecoglossicida, AWI5 belongs to P. lini, ABI12, ABI15 and ABI16 belong to P. taiwanensis. Moreover, the bacterial efficacy at a suspension concentration of 0.5 OD (80 %) was significantly greater than that at a concentration of 0.2 OD (63.33 %). No significant difference was detected in the response of the pest larvae or eggs to the different strains. Furthermore, olfactory trials revealed that the female parasitoid wasp Habrabracon hebetor actively avoided the infection of the treated larvae by the strains. These findings have practical implications for the development of microbiological pest control strategies.

Genomic, phylogenetic and catabolic re-assessment of the Pseudomonas putida clade supports the delineation of Pseudomonas alloputida sp. nov., Pseudomonas inefficax sp. nov., Pseudomonas persica sp. nov., and Pseudomonas shirazica sp. nov.

Bacteria of the Pseudomonas putida group are studied for a large panel of properties ranging from plant growth promotion and bioremediation to pathogenicity. To date, most of the classification of individual pseudomonads from this group relies on 16S RNA gene analysis, which is insufficient for accurate taxonomic characterization within bacterial species complexes of the Pseudomonas putida group. Here, a collection of 20 of these bacteria, isolated from various soils, was assessed via multi-locus sequence analysis of rpoD, gyrB and rrs genes. The 20 strains clustered in 7 different clades of the P. putida group. One strain per cluster was sequenced and results were compared to complete genome sequences of type strains of the P. putida group. Phylogenetic analyses, average nucleotide identity data and digital DNA hybridizations, combined to phenotypic characteristics, resulted in the proposition and description of four new species i.e. Pseudomonas alloputida Kh7 T (= LMG 29756 T = CFBP 8484 T) sp. nov., Pseudomonas inefficax JV551A3 T (= DSM108619 T = CFBP 8493 T) sp. nov., Pseudomonas persica RUB6 T (= LMG 29757 T = CFBP 8486 T) sp. nov. and Pseudomonas shirazica VM14 T (= LMG 29953 T = CFBP 8487 T) sp. nov.

Phylogenetic diversity and antagonistic traits of root and rhizosphere pseudomonads of bean from Iran for controlling Rhizoctonia solani.

Fluorescent pseudomonads from bean root and rhizosphere in Iran were investigated for biocontrol of the fungal pathogen Rhizoctonia solani. Phylogenetic analysis of concatenated 16S rRNA, gyrB and rpoD sequences for 33 Pseudomonas isolates showed that 15 belonged to four clusters within the 'P. fluorescens' group, i.e. one corresponding to P. thivervalensis, two others including P. moraviensis or P. baetica, and the last one without closely-related established species. The 18 other isolates belonged to five clusters within the 'P. putida' group, one including P. mosselii and P. entomophila, another including strains currently described as P. putida, and three without closely-related species described. Ten isolates were selected based on in vitro inhibition of R. solani. Cellulase activity was identified in three pseudomonads, chitinase activity in two pseudomonads, extracellular protease activity in nine pseudomonads and hydrogen cyanide production in two pseudomonads. Genes coding for production of phenazine, pyoluteorin, pyrrolnitrin and 2,4-diacetylphloroglucinol were not found, whereas the 1-aminocyclopropane-1-carboxylate deamination gene acdS was present in three pseudomonads. The antagonistic acdS+ strain VKh13 from the 'P. putida' group effectively protected soil-grown bean from R. solani AG 4-HGI. Results show that pseudomonads from uncharacterized taxa were readily obtained from Iranian soils and displayed biocontrol potential against R. solani.

The role of nitric oxide in basal and induced resistance in relation with hydrogen peroxide and antioxidant enzymes.

Nitric oxide (NO) is one of the main signal molecules, which is involved in plant growth and development and can change regular physiological activity in biotic and abiotic stresses. In this study, the role of NO in induced resistance with Pseudomonas fluorescent (CHA0) and basal resistance against Rhizoctonia solani in bean plant was investigated. Our results revealed that P. fluorescent and R. solani can increase NO production at 6h post inoculation (hpi). Also, using the NO donor S-nitroso-N-acetyl D-penicillamine (SNAP) led to increase NO and bean plant resistance against R. solani. Utilizing the NO scavenger, 2-(4-carboxyphenyl)-4,4,5,5-tetramethy-limidazoline-1-oxyl-3-oxide (cPTIO), not only decreased basal resistance but also reduced induced resistance. In continue, the activity of antioxidant enzymes was studied in the former treatments. SNAP, CHA0 and R. solani increased the activity of peroxidase (POX), catalase (CAT) and ascorbate peroxidase (APX) at 6, 12 and 24h post inoculation (hpi). In contrast, using cPTIO and R. solani simultaneously (cPTIO+R) showed reduction in activity of POX and APX at 6 hpi. The cPTIO+R treatment increased POX, APX and CAT activity at 12 and 24 hpi. Hydrogen peroxide (H2O2) monitoring in the leaf discs clarified that SNAP can increase H2O2 production like CHA0 and R. solani. On the other hand, SNAP increased the resistance level of leaf discs against R. solani. Treating the leaf discs with cPTIO led to decrease resistance against the pathogen. These leaf discs showed reduction in H2O2 production at 6 hpi and suddenly enhanced H2O2 generation was observed at 24hpi. This study showed that CHA0 can increase NO level in bean plants. NO induced H2O2 generation and regulated redox state of the host plant. This interaction resulted in significant defense against the pathogen.