Root damage to apple plants by cockchafer larvae induces a change in volatile signals below‐ and above‐ground

Volatile organic compounds (VOCs) mediate communication between plants and insects. Plants under insect herbivore attack release VOCs either at the site of attack or systemically, indicating within‐plant communication. Some of these VOCs, which may be induced only upon herbivore attack, recruit parasitoids and predatory insects to feed on the attacking insects. Moreover, some plants are able to ‘eavesdrop’ on herbivore‐induced plant volatiles (HIPVs) to prime themselves against impending attack; such eavesdropping exemplifies plant–plant communication. In apple orchards, the beetle Melolontha melolontha L. (Coleoptera: Scarabaeidae) is an important insect pest whose larvae live and feed on roots for about 4 years. In this study, we investigated whether the feeding activity of M. melolontha larvae (1) alters the volatile profile of apple roots, (2) induces the release of HIPVs systemically in the leaves, and (3) whether infested plants communicate to neighbouring non‐infested conspecifics through HIPVs. To answer these questions, we collected constitutive VOCs from intact M9 roots as well as M. melolontha larvae‐damaged roots using a newly designed ‘rhizobox’, to collect root‐released volatiles in situ, without damaging the plant root system. We also collected VOCs from the leaf‐bearing shoots of M9 whose roots were under attack by M. melolontha larvae and from shoots of neighbouring non‐infested conspecifics. Gas chromatography‐mass spectrometry analysis showed that feeding activity of M. melolontha larvae induces the release of specific HIPVs; for instance, camphor was found in the roots only after larvae caused root damage. Melolontha melolontha also induced the systemic release of methyl salicylate and (E,E)‐α‐farnesene from the leaf‐bearing shoots. Methyl salicylate and (E,E)‐α‐farnesene were also released by the shoots of non‐infested neighbouring conspecifics. These phenomena indicate the induction of specific VOCs below‐ and above‐ground upon M. melolontha larvae feeding on apple roots as well as plant–plant communication in apple plants.     

Treatment of millet crop plant (Sorghum bicolor) with the entomopathogenic fungus (Beauveria bassiana) to combat infestation by the stem borer,Chilo partellus Swinhoe (Lepidoptera: Pyralidae)

Experiments were done to test if Beauveria bassiana can become an endophyte in sorghum and confer protection from stem borer. Four-week-old sorghum seedlings were treated with B. bassiana. The plants were examined for endophytic presence of B. bassiana, 30 and 60 days after treatment. Stem cultures from treated plants showed growth of B. bassiana. PCR amplification using fungal specific primers for a conserved region of β tubulin gene yielded identical 360 bp products from both B. bassiana and treated sorghum plants. In a subsequent experiment, B. bassiana treated and untreated (control) sorghum plants were artificially infested with stem borer (Chilo partellus) larvae 15 days post treatment and the extent of damage was compared. About 40% of the control plants developed dead heart while no plant in the B. bassiana treated plot did. In the surviving control plants, stem tunneling by shoot borer was significantly higher compared to B. bassiana treated sorghum plants.     

Host‐plant associated genetic divergence of two Diatraea spp. (Lepidoptera: Crambidae) stemborers on novel crop plants

Diatraea lineolata and Diatraea saccharalis (Lepidoptera: Crambidae) are moths with stemboring larvae that feed and develop on economically important grasses. This study investigated whether these moths have diverged from a native host plant, corn, onto introduced crop plants including sorghum, sugarcane, and rice. Diatraea larvae were collected from these four host plants throughout the year in El Salvador and were reared on artificial diet until moths or parasitoids emerged. Adult moths were subsequently identified to species. Amplified fragment length polymorphisms (AFLPs) and mitochondrial DNA cytochrome oxidase I (COI) were used to examine whether or not there was genetic divergence of D. lineolata or D. saccharalis populations on the four host plants. Percent parasitism was also determined for each moth on its host plants. D. lineolata was collected from corn in the rainy season and sorghum in the dry season. D. saccharalis was most abundant on sugarcane in the rainy season and sorghum in the dry season. The AFLP analysis found two genetically divergent populations of both D. lineolata and D. saccharalis. Both moths had high levels of parasitism on their dominant host plant in the rainy season, yet had low levels of parasitism on sorghum in the dry season. The presence of two genotypes of both Diatraea spp. on sorghum suggest that host‐associated differentiation is occurring on this novel introduced crop plant.     

Genetic population structure of sugarcane aphid,Melanaphis sacchari,in sorghum,sugarcane, and Johnsongrass in the continental USA

In 2013, an outbreak of Melanaphis sacchari Zehntner (Hemiptera: Aphididae) was reported in sorghum in Texas, USA. Although this aphid has been reported in the continental USA for nearly a century, its occurrence was limited to Florida and Louisiana sugarcane. After 2013 and within just 3 years M. sacchari was reported in almost all sorghum growing regions from south central to southeastern states in the USA. Sorghum fields in affected areas have sustained considerable losses. This aphid has also been reported on Johnsongrass and other feral grasses. The speed at which this aphid has spread raises serious concerns about future infestations. Many aphid species present genetically distinct populations when feeding on different host plants. Thus, it was hypothesized that the recent outbreak in sorghum could be explained by a recent introduction of a sorghum‐specialized genotype. In this study, we genetically characterized M. sacchari in three of its most common host plants – sorghum, sugarcane, and Johnsongrass – across its geographic distribution in the continental USA. Although M. sacchari specimens were grouped within three genetically distinct clusters, we did not find evidence of host plant or geographic population structure. Our characterization of the genetic structure of this pest provides baseline data aimed to help explain its recent outbreak in sorghum, as well as information that may aid in the design of sustainable control strategies.     

Fusarium mirum sp. nov,intertwining Fusarium madaense and Fusarium andiyazi,pathogens of tropical grasses

Many species in the Fusarium fujikuroi Species Complex (FFSC) have an affinity for grass species, with whom they live in an endophytic association or cause disease. We recovered isolates of Fusarium from agriculturally important grasses in Africa and Brazil, and characterized them with morphological markers, mating type, and Amplified Fragment Length Polymorphisms (AFLPs). We also conducted multi-locus phylogenetic analyses based on partial DNA sequences of translation elongation factor-1α (TEF1), β-tubulin (TUB), and the second largest subunit of RNA polymerase (RPB2) gene regions. Sexual cross fertility was used to test the biological species concept and the sexual stage of F. madaense is described. A novel species within the FFSC, Fusarium mirum, that is different from the other known species in the complex, was formally described. Fusarium mirum, F. madaense, and Fusarium andiyazi are a tightly intertwined species trio that are morphologically identical, but phylogenetically distinguishable, and amongst whom interspecific genetic exchange may still occur. These three species are so close that they cannot be reliably distinguished if only sequences of the TEF1 gene are used. In pathogenicity tests, all tested isolates of F. madaense from sugarcane, sorghum, maize, millet and Brachiaria could induce stalk rot in sorghum, maize and millet, and pokkah boeng in sugarcane. This study increases our understanding of the diversity of species within the FFSC that cause disease in tropical grasses or act as endophytes, and their geographic distributions. The genetically close relationship between F. mirum, F. madaense, and F. andiyazi provides an opportunity to study and identify factors underlying their limited inter-specific cross-fertility and sympatric speciation.     

Ethanol production potential of sweet sorghum assessed using forage fiber analysis procedures

Sweet sorghum (Sorghum bicolor (L.) Moench) is widely recognized as a highly promising biomass energy crop with particular potential to complement sugarcane production in diversified cropping systems. Agronomic assessments have led to identification of four cultivars well suited for such sugarcane‐based production systems in southern Louisiana. Sweet sorghum biofuel production systems are currently being developed, and research producing large sample numbers requiring ethanol yield assessment is anticipated. Fiber analysis approaches developed for forage evaluation appear to be useful for screening such large numbers of samples for relative ethanol yield. Chemical composition, forage fiber characteristics, digestibility, and ethanol production of sweet sorghum bagasse from the four cultivars were assessed. Measures of detergent fiber, lignin, and digestibility were highly correlated with ethanol production (P < 0.01). The best linear regression models accounted for about 80% of the variation among cultivars in ethanol production. Bagasse from the cultivar Dale produced more ethanol per gram of material than any of the other cultivars. This superior ethanol production was apparently associated with less lignin in stems of Dale. Forage evaluation measures including detergent fiber analyses, in vitro digestibility, and an in vitro gas production technique successfully identified the cultivar superior in ethanol yield indicating their usefulness for screening sweet sorghum samples for potential ethanol production in research programs generating large sample numbers from evaluations of germ plasm or agronomic treatments. These screening procedures reduce time and expense of alternatives such as hexose sugar assessment for calculating theoretical ethanol yield.     

Expression of soybean proteinase inhibitors in transgenic sugarcane plants: effects on natural defense against Diatraea saccharalis

The introduction and expression of proteinase inhibitor encoding genes into sugarcane (Saccharum officinarum L.) genome is an interesting strategy for conferring partial resistance to the sugarcane borer Diatraea saccharalis (Lepidoptera: Crambidae), the major insect pest of sugarcane in Brazil. To investigate the role of soybean (Glycine max L.) Kunitz trypsin inhibitor (SKTI) and soybean Bowman–Birk inhibitor (SBBI) in the control of D. saccharalis, the cDNAs encoding these proteinase inhibitors were placed under the control of the maize ubiquitin promoter (Ubi-1), and introduced into sugarcane callus using particle bombardment. Putative transgenic plants were initially identified after regeneration from callus growing in the presence of 30 mg l^–1 geneticin, while molecular characterization of transgenic plants revealed that both genes were incorporated into the sugarcane genome and expressed. We also carried out insect feeding trials using D. saccharalis neonate larvae and leaf tissue excised from propagated transgenic and untransformed plants, and found that the growth of larvae feeding on leaf tissue from transgenic plants containing BBI and Kunitz inhibitors was significantly retarded as compared to larvae fed on leaf tissue from untransformed plants. In greenhouse trials with transgenic sugarcane plants infested with D. saccharalis neonates, we found that these plants still presented the ‘dead heart’ symptom typically observed in susceptible plants in the field, suggesting that the retardation of the growth of D. saccharalis observed in the laboratory-based feeding trials was not sufficient to prevent this type of damage.     

Comparison of volatile organic compounds from uninfested and Monochamus alternatus Hope infested Pinus massoniana Lamb.

Monochamus alternatus is a destructive stem‐boring herbivore of Pinus massoniana, and the principal vector of pine wood nematode. To investigate the impacts of boring by M. alternatus larvae on the emission of volatile organic compounds (VOCs) from their host trees, the VOCs from uninfested and M. alternatus larvae infested P. massoniana trees were observed using a gas chromatograph–mass spectrometer. We detected 12, 9, 18 and 14 volatile organic compounds from infested xylem, infested phloem, uninfested xylem and uninfested phloem, respectively. In P. massoniana xylem, the boring of M. alternatus larvae induced cyclosativene, and inhibited 4‐carene, humulene, styrene, α‐phellandrene, β‐myrcene, β‐phellandrene and γ‐terpinene. The relative amounts of camphene, copacamphene, longicyclene, longifolene, tricyclene and α‐longipinene were significantly increased, and the relative amounts of α‐pinene and β‐pinene were significantly decreased by the boring behaviors of M. alternatus larvae. In P. massoniana phloem, the boring of M. alternatus larvae induced 2‐bornanone, copacamphene, longicyclene and α‐longipinene, and inhibited 2‐carene, 4‐carene, styrene, α‐phellandrene, β‐myrcene, β‐phellandrene, β‐pinene, γ‐terpinene and ο‐cymene. The relative amounts of camphene, caryophyllene and longifolene were significantly increased by the boring behaviors of M. alternatus larvae. The results indicate that the boring behaviors of M. alternatus larvae changed both the sorts and contents of the VOCs from P. massoniana trees.     

Genetic and morphological comparisons ofGlomerella (Colletotrichum) isolates from maize and from sorghum

Various morphological and genetic characteristics were compared among six isolates ofColletotrichum from maize and six from sorghum. For the first time, a teleomorph was induced in sorghum isolates by pairing them on autoclaved sorghum leaves in a humidity chamber. The sorghum teleomorph was morphologically similar toGlomerella graminicola andGlomerella tucumanensis, the teleomorphs ofColletotrichum isolates from maize and from sugarcane, respectively. Mating tests demonstrated thatGlomerella isolates from maize and sorghum were not interfertile. Several small but consistent differences in the morphologies of the isolates from maize and from sorghum were observed which agreed with earlier reports. DNA fingerprints detected as restriction fragment length polymorphisms of mitochondrial DNA and random polymorphic DNA (RAPD) produced from nuclear DNA by the polymerase chain reaction could be used to reliably and unambiguously distinguish members of the two groups of isolates. Results of a statistical analysis of similarity of the RAPD fingerprints suggested that maize and sorghum isolates ofColletotrichum are only about 45% similar (±10%) and represent two distinct and separate genetic lineages. We conclude that isolates ofColletotrichum from maize and sorghum are sibling species since they are morphologically very similar but reproductively completely isolated.     

Analysis of volatile organic compounds in rats with dopaminergic lesion: Possible application for early detection of Parkinson’s disease

Parkinson’s disease (PD) is characterized by dopaminergic (DA) neuron depletion. Early detection of PD may help in selecting the appropriate treatment. Biomarkers of PD have been suggested, however none of these is currently in clinical use. The aim of this study was to identify volatile organic compounds (VOCs) as early biomarkers of PD. Our hypothesis was that during PD progression, specific VOCs are generated that are linked to the biochemical pathways characterizing PD. These VOCs can be detected by GC–MS combined with solid-phase microextraction (SPME) technique. Three groups of rats were studied: DA-lesioned rats injected with 6-hydroxydopamine (HDA; 250 μg/rat n = 11); control rats injected with saline (n = 9), and control rats injected with DSP-4 (n = 8), a specific noradrenergic neuron toxin. Blood and striatal tissue homogenate were analyzed. In the blood, 1-octen-3-ol and 2-ethylhexanol were found at significantly higher concentrations in HDA versus sham rats. In the striatal homogenate 1-octen-3-ol and other four compounds were found at significantly lower concentrations in HDA versus sham rats. 1-Octen-3-ol is a cytotoxic compound. These results may lead to the development of an early diagnostic test for PD based on profiling of VOCs in body fluids.     

Construction of a composite sorghum genome map and comparison with sugarcane, a related complex polyploid

 A sorghum composite linkage map was constructed with two recombinant inbred line populations using heterologous probes already mapped on maize and sugarcane. This map includes 199 loci revealed by 188 probes and distributed on 13 linkage groups. A comparison based on 84 common probes was performed between the sorghum composite map and a map of a sugarcane (Saccharum spp.) cultivar being developed and presently comprising 10 tentative linkage groups. A straight synteny was observed for 2 pairs of linkage groups; in two cases, 1 sorghum linkage group corresponded to 2 or 3 sugarcane linkage groups, respectively; in two cases 1 sugarcane link- age group corresponded to 2 separate sorghum linkage groups; for 2 sorghum linkage groups, no complete correspondance was found in the sugarcane genome. In most cases loci appeared to be colinear between homoeologous chromosomal segments in sorghum and sugarcane. These results are discussed in relation to published data on sorghum genomic maps, with specific reference to the genetic organization of sugarcane cultivars, and they, illustrate how investigations on relatively simple diploid genomes as sorghum will facilitate the mapping of related polyploid species such as sugarcane. Received: 12 August 1996 / Accepted: 30 August 1996     

On-line detection of root-induced volatiles in Brassica nigra plants infested with Delia radicum L. root fly larvae

Plants emit various volatile organic compounds (VOCs) upon herbivore attack. These VOC emissions often show temporal dynamics which may influence the behavior of natural enemies using these volatiles as cues. This study analyzes on-line VOC emissions by roots of Brassica nigra plants under attack by cabbage root fly larvae, Delia radicum. Root emitted VOCs were detected using Proton-Transfer-Reaction Mass Spectrometry (PTR-MS) and Gas Chromatography–Mass Spectrometry (GC–MS). These analyses showed that several sulfur containing compounds, such as methanethiol, dimethyl sulfide (DMS), dimethyl disulfide (DMDS), dimethyl trisulfide (DMTS) and glucosinolate breakdown products, such as thiocyanates (TC) and isothiocyanates (ITC), were emitted by the roots in response to infestation. The emissions were subdivided into early responses, emerging within 1–6 h after infestation, and late responses, evolving only after 6–12 h. The marker for rapid responses was detected at m/z 60. The ion detected at m/z 60 was identified as thiocyanic acid, which is also a prominent fragment in some TC or ITC spectra. The emission of m/z 60 stopped when the larvae had pupated, which makes it an excellent indicator for actively feeding larvae. Methanethiol, DMS and DMDS levels increased much later in infested roots, indicating that activation of enzymes or genes involved in the production of these compounds may be required. Earlier studies have shown that both early and late responses can play a role in tritrophic interactions associated with Brassica species. Moreover, the identification of these root induced responses will help to design non-invasive analytical procedures to assess root infestations.     

Arabidopsis thaliana as a model system for testing the effect of Trichoderma volatile organic compounds

In ecosystems, plant and bacterial volatile organic compounds (VOCs) are known to influence plant growth but less is known about the physiological effects of fungal VOCs. We have used Arabidopsis thaliana as a model to test the effects of VOCs from the soil fungus Trichoderma viride. Mature colonies of T. viride cultured on Petri plates were placed in a growth chamber in a shared atmosphere with A. thaliana without direct physical contact. Compared to controls, plants grown in the presence of T. viride volatiles were taller, bigger, flowered earlier, and had more lateral roots. They also had increased total biomass (45 %) and chlorophyll concentration (58 %). GC–MS analysis of T. viride VOCs revealed 51 compounds of which isobutyl alcohol, isopentyl alcohol, and 3-methylbutanal were most abundant. We conclude that VOCs emitted by T. viride have growth promoting effects on A. thaliana in the absence of direct physical contact.     

Production of volatile organic compounds by Aureobasidium pullulans as a potential mechanism of action against postharvest fruit pathogens

Two Aureobasidium pullulans strains (L1 and L8), effective against some fruit postharvest pathogens were evaluated for VOCs production as a part of their modes of action towards five pathogens (Botrytis cinerea, Colletotrichum acutatum, Penicillium expansum, Penicillium digitatum and Penicillium italicum). The VOCs were assayed with a double petri dish assay against conidia germination of target pathogens. Results obtained showed that the VOCs generated by the antagonists inhibited significantly the conidia germination of all pathogens compared to the control. In particular, the conidia germination of all Penicillium was completely inhibited by VOCs produced by L1 and L8. In in vivo tests, apples and oranges were artificially inoculated with pathogen conidia and then biofumigated with VOCs emitted by both antagonists. The antagonistic treatment controlled significantly pathogen infection, confirming the results obtained in vitro tests. The best L1 and L8 VOCs activity was observed on apple inoculated with B. cinerea where the lesion diameter reduction observed was greater than the 88%. The compounds emitted by L1 and L8 strains were identified with the solid-phase microextraction (SPME)–gas chromatographic technique. Compounds as 2-phenyl, 1-butanol-3-methyl, 1-butanol-2-methyl and 1-propanol-2-methyl belonging to the group of alcohols were mainly produced for both strains, in the first 96 h of growth. These compounds were confirmed by comparison with standards. The pure compounds of VOCs cited above were used to determine the EC₅₀ values for conidia germination of pathogens. The 1-propanol-2-methyl was the VOC least active against all tested fungi, with the EC₅₀ values over 0.8 μl ml⁻¹, while the 2-phenethyl alcohol was the most active with EC₅₀ values lower than 0.8 μl ml⁻¹, except for the C. acutatum (1.97 μl ml⁻¹). The present study demonstrated, for the first time, that the production of VOCs could play an essential role in the antagonistic activity of two A. pullulans strains against five fruit postharvest pathogens.     

A new species and two new Chinese records of Ochroconis from sugarcane and banana rhizosphere in Guangxi,China

Ochroconis guangxiensis isolated from sugarcane and banana rhizosphere, was described as a new species based on morphological characteristics and phylogenetic analysis using sequence data of the nuclear small subunit rRNA gene (SSU), internal transcribed spacer (ITS) region and large subunit (LSU) rRNA gene. Taxonomic and phylogenetic remarks are also provided for O. minima and O. ramosa. The latter two species are newly recorded for China. These three Ochroconis species, as dark septate endophytes, inhabit rhizosphere and can form a symbiosis with sugarcane and banana.     

Aphid antixenosis in cotton is activated by the natural plant defence elicitor cis-jasmone

Upon insect herbivory, plants can release blends of volatile organic compounds (VOCs) that modify herbivore and natural enemy behaviour. We have shown recently that cotton, Gossypium hirsutum, emits a blend of defence VOCs that repels the cotton aphid, Aphis gossypii, upon herbivory by this notorious crop pest, including (Z)-3-hexenyl acetate, (E)-4,8-dimethyl-1,3,7-nonatriene (DMNT), methyl salicylate and (E,E)-4,8,12-trimethyl-1,3,7,11-tridecatetraene (TMTT). In this study, we investigated changes in the defence VOC profile of G. hirsutum induced by the naturally-occurring plant elicitor cis-jasmone (CJ) and whether these changes modify the behaviour of A. gossypii. In four-arm olfactometer assays, VOCs from untreated plants were significantly attractive (P < 0.05), whilst VOCs from CJ-treated plants were significantly repellent (P < 0.05). The VOCs induced by CJ appeared to comprise (Z)-3-hexenyl acetate, DMNT, methyl salicylate and TMTT. In quantitative VOC collection studies, sustained release of DMNT and TMTT was observed in CJ-treated plants over a period of five days, with levels becoming statistically significantly higher than for control treated plants on the fifth day in most cases. Despite earlier indications, no statistically significant differences were observed in levels of (Z)-3-hexenyl acetate or methyl salicylate between CJ and control treatments on any day. Furthermore, DMNT and TMTT emissions from CJ-treated plants were further enhanced by subsequent addition of A. gossypii. CJ treatment induced statistically significantly higher DMNT and TMTT expression levels as early as day three, when A. gossypii was present. The results in this study show that CJ can induce the production of A. gossypii-induced VOCs from G. hirsutum, with potential for deployment in novel crop protection strategies.     

Enhancement of exopolysaccharide production from Ganoderma lucidum using a novel submerged volatile co-culture system

Based on the impact of volatile organic compounds (VOCs) on secondary metabolite pathways, a novel submerged volatile co-culture system was constructed, and the effects of thirteen fungal and bacterial VOCs were investigated on Ganoderma lucidum exopolysaccharides production. The results demonstrated at least a 2.2-fold increase in exopolysaccharide (EPS) specific production yield in 6 days submerged volatile co-culture of G. lucidum with Pleurotus ostreatus. Therefore, P. ostreatus was selected as a variable culture, and the effects of agitation speed, inoculum size, initial pH, and co-culture volume on EPSs production were investigated using a Taguchi L₉ orthogonal array. Finally, the highest concentration of EPSs (3.35 ± 0.22 g L^?1) was obtained under optimized conditions; initial pH 5.0, inoculum size 10%, 150 rpm, and 3:1 volume ratio of variable culture to main culture.     

Pseudomonas spp. Mediate defense response in sugarcane through differential exudation of root phenolics

Pseudomonas spp., a ubiquitous biocontrol agent, protects the plants from phytopathogens by suppressing them directly by reinforcing the plant’s intrinsic defense mechanism. Root exudated phenolics play an important role in establishing the rhizobacteria population and cross the host boundaries in beneficial plant–microbe interaction. In this study, Pseudomonas spp. HU-8 & HU-9 antagonized the sugarcane red rot pathogen (C. falcatum) and showed a positive chemotactic response against different concentrations (10–30 µM) of synthetic phenolic acids like p-coumaric, vanillic, and 3,4 di-hydroxybenzoic acid. In a pot experiment, they effectively colonized the sugarcane rhizosphere and mediated defense response in sugarcane plants challenged with red rot pathogen C. falcatum by regulating the exudation of root phenolics under hydroponic conditions. They significantly induced the activity of the antioxidant enzymes CAT (1.24–1.64 fold), PO (0.78–1.61 fold), PAL (0.77–0.97 fold), and PPO (3.67–3.73 fold) over untreated plants in sugarcane. They also induced the total phenolic contents (TPC) in sugarcane in the presence (6.56–10.29 mg/g GAE) and absence (2.89–4.16 mg/g GAE) of the pathogen quantified through the Folin-Ciocalteu (FC) method. However, their effect was lower than that of the pathogen (4.34–8 mg/g GAE). The Pseudomonas spp. significantly colonized the sugarcane rhizosphere by maintaining a cell population of (1.0E + 07–1.3E + 08 CFU/mL). A significant positive Pearson’s correlation was observed between the root exudated total phenolic contents, antioxidant enzymatic activities, and rhizospheric population of inoculated bacteria. The 16S rRNA and rpoD gene analysis showed sequence conservation (C: 0.707), average number of nucleotide differences (k: 199.816), nucleotide diversity, (Pi): 0.09819), average number of informative nucleotide sites per site (Psi: 0.01275), GC content (0.57), and polymorphic sites (n = 656). These diverse Pseudomonas spp. could be an ideal bio-inoculants for a broad range of hosts especially graminaceous crops.