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Background: Brucellosis is an economically important zoonotic disease caused by the gram negative bacteria belonging to the genus Brucella. Medicinal plants are well known for a wide variety of potential antimicrobial agents that can be used as anti-microbial drugs. Method: In the present study, crude ethanol and methanol extracts of local plants (Berberies lyceum and Fagonia cretica) were tested in vitro against Brucella melitensis via well diffusion method for their antibacterial activity. In in-silico study, phytochemicals previously identified in the selected plants were docked with homology model of the cytotoxic factor malate synthase G (MSG) highly conserved among Brucella spp., in Molecular Operating Environment (MOE) to predict a potential drug against B. melitensis. Molecular dynamic simulation was performed to predict the stability of MSG through MOE. Result: Ethanolic crude extracts of B. lyceum showed maximum zone of inhibition (32.5 mm) followed by methanolic extracts (30 mm), while ethanolic extracts of F. cretica showed zone of inhibition (29 mm) followed by methanolic extracts (27.5 mm). In silico screening predicted phytic acid as the most potent inhibitor followed by jehlumine, barbamine, oxyberberine and sindamine. Conclusion: The synergistic utilization of phytochemicals derived from B. lyceum may potentially provide protection against B. melitensis.

Chemical crosstalk between plant roots and associated microbes is imperative for the initiation of symbiosis. Phenolics, flavonoids and indole-3-acetic acid (IAA) are acclaimed as signaling molecules in developing plant-microbe symbiosis. The role of IAA and flavonoids in establishing a symbiotic relationship between Aspergillus nomius wlg2 and maize roots have also been observed during the present study. The isolate A. nomius wlg2 and maize roots have secreted flavonoids and IAA that helped in A. nomius-maize association. However, the suppression of either flavonoids or IAA secretion reduced the endophyte colonization of the maize roots by 77% and 67%, respectively. The main flavonoids secreted by maize were identified as luteolin, calycosin, phenolics cis-caftaric acid, and caffeoyl-d-glucose. The identified flavonoids were most likely to be involved in the root-endophyte association. To establish a plant-microbe interaction, the complex chemical dialogue is necessary for which IAA and flavonoids play a central role.

Different physical and chemical techniques are used for the decontamination of Cr+6 contaminated sites. The techniques are expensive, laborious, and time-consuming. However, remediation of Cr+6 by microbes is viable, efficient, and cost-effective. In this context, plant growth-promoting rhizobacteria Acinetobacter bouvetii P1 isolated from the industrial zone was tested for its role in relieving Cr+6 induced oxidative stress in sunflower. At the elevated Cr+6 levels and in the absence of P1, the growth of the sunflower plants was inhibited. In contrast, the selected strain P1 restored the sunflower growth under Cr+6 through plant growth–promoting interactions. Specifically, P1 biotransformed the Cr+6 into a stable and less toxic Cr+3 form, thus avoiding the possibility of phytotoxicity. On the one hand, the P1 strengthened the host antioxidant system by triggering higher production of enzymatic antioxidants, including catalases, ascorbate peroxidase, superoxide dismutase, and peroxidase. Similarly, P1 also promoted higher production of nonenzymatic antioxidants, such as flavonoids, phenolics, proline, and glutathione. Apart from the bioremediation, P1 solubilized phosphate and produced indole acetic acid, gibberellic acid, and salicylic acid. The production of phytohormones not only helped the host plant growth but also mitigated the harsh condition posed by the elevated levels of Cr+6. The findings mentioned above suggest that P1 may serve as an excellent phyto-stimulant and bio-remediator in a heavy metal-contaminated environment.

Plants and trees make our cities more attractive and provide many ecosystem services but some of these also cost society harm in the form of pollen allergy. According to an estimate, 60-70 percent of the trees in Islamabad are of Paper mulberry. Thirty percent of the people are affected each year from pollen allergy in Islamabad. This study evaluated age and gender-wise effects of Pollen Allergy on health followed by proposing options for Paper Mulberry as an alternative. The economic and environmental costs and benefits of Paper mulberry and its alternative plants were also quantified. The annual benefits and costs of Paper Mulberry and its alternative plants were estimated through “The National Tree Benefits Calculator”. Descriptive statistics along with expert opinion method was used for the analysis of the data. The pollen allergy affects children more as compared to other age groups and more on female as compared to male. Urban respondents were affected more by pollen allergy as compared to rural respondents. Most of the respondents were found affected in the month of March. The average monthly health expenditures of the respondents from pollen as Rs. 879. Pine Rexburg and Kachnar are viable alternatives for Paper Mulberry from both economic and environmental viewpoints. Based on the findings it is concluded that Pine Rexburg and Kachnar are the best alternative for Paper Mulberry in Islamabad. The Capital Development Authority should replace Paper Mulberry with Pine Rexburg and Kachnar in the city. Besides, Government should arrange awareness program especially in the pollen month (March) and should provide free of cost treatment and preventives for the pollen patients.

Plants can be severely affected by heat stress due to rapid environmental changes. Use of endophytic fungi is a new tool to protect crops from the environmental stresses. Here we claim a potent endophyte isolated from Sonchus asper L. The aim was to explore the stress adaptive mechanism of sunflower and soybean mediated by Aspergillus niger (SonchL-7) under high temperature. Inoculation with A. niger boosted plant height, biomass and chlorophyl contents, while significantly curtailed the concentration of lipid peroxidation and reactive oxygen species (ROS) under thermal stress at 40°C. Moreover, the ROS-scavenging activities, like ascorbic acid oxidase (AAO), catalase (CAT), glutathione reductase (GR), superoxide dismutase (SOD) and peroxidase (POD) were augmented. Also, proline and phenolics were enhanced in the tested crops, while ABA concentration was significantly reduced. These positive results suggested that A. niger can be used as a heat-stress ameliorative tool for crops in the future.

The growth promoting activities of the isolated endophyte Aspergillus terreus from Aloe barbendsis was studied in the salt stressed Pennisetum glaucum (pearl millet). A significant (P = 0.05) increase in the root-shoot lengths, fresh and dry weights and chlorophyll content of pearl millet seedlings was noticed after colonization by A. terreus under normal conditions. At 100 mM NaCl stress and A. terreus inoculation, the growth rate of pearl millet seedlings were significantly (P = 0.05) inhibited. Furthermore, the IAA production, relative water content (RWC), chlorophyll, soluble sugar, phenol and flavonoid contents were significantly decreased, whereas proline content and lipid peroxidation were increased. On the contrary, pearl millet seedlings inoculated with A. terreus retained significantly (P = 0.05) higher amounts of RWC, chlorophyll, soluble sugar, phenol and flavonoid contents under 100 mM salt stress. The higher IAA production in A. terreus associated seedlings rescued the plant growth and development under salt stress. Moreover, the LC MS/MS analysis of A. terreus cultural filtrate revealed the presence of quinic acid, ellagic acid, calycosin, wogonin, feruloylquinic acid, caffeic acid phenylethyl ester, D-glucoside, myricetin, propoxyphene and aminoflunitrazepam. The results of the study conclude that innoculation of A. terreus improves the NaCl tolerance in pearl millet by ameliorating the physicochemical attributes of the host plants.

Salinity stress can severely affect the growth and production of the crop plants. Cheap and reliable actions are needed to enable the crop plants to grow normal under saline conditions. Modification at the molecular level to produce resistant cultivars is one of the promising, yet highly expensive techniques, whereas application of endophytes on the other hand are very cheap. In this regard, the role of Cochliobolus sp. in alleviating NaCl-induced stress in okra has been investigated. The growth and biomass yield, relative water content, chlorophyll content and IAA were decreased, whereas malondialdehyde (MDA) and proline content were increased in okra plants treated with 100 mM NaCl. On the contrary, okra plants inoculated with C. lunatus had higher shoot length, root length, plant dry weight, chlorophyll, carotenoids, xanthophyll, phenolicss, flavonoids, IAA, total soluble sugar and relative water content, while lower MDA. LC-MS/MS analysis of the Cochliobolus sp. extract revealed the presence of pinocembrin, chlorogenic acids, wogonin, calycosin and diadzein as a salinity stress reliever. From the results, it can be concluded that colonization of Cochliobolus sp. improves the NaCl tolerance by ameliorating the physicochemical attributes of the host plants.

Auxin is the reciprocal signaling molecule, which interferes with other phyto-hormonal and physiological processes during plant–microbes interaction. In this regard, Bipolaris spp., a growth-promoting endophytic fungus was used to inoculate pre-stressed Zea mays seedlings with yucasin (IAA inhibitor). The IAA-deficient host was heavily colonized by the endophyte that subsequently promoted the host growth and elevated the IAA levels with a peak value at 72 h. However, the seedling growth was inhibited later (i.e., at 120 h) due to the high levels of IAA that interfered with the activity of phytoalexins and brassinosteroids. Such interference also modulated the endophytic fungus from symbiotic to biotrophic pathogen that left the host plants defenseless.

Plants are susceptible to various environmental constrains, including heat stress due to their sessile nature. Endophytic fungi can be used as a novel technique to protect crop plants against the injurious effects of thermal stress. Endophytic fungi were isolated from Adiantum capillus-veneris L. and tested against heat stress in Glycine max L. and Helianthus annuus L. The results exhibited increased levels of the plant’s chlorophyll, height and biomass in Aspergillus foetidus (AdR-13) inoculated host crop species. Conversely, a significant decrease in lipid peroxidation and reactive oxygen species (ROS) production was noted in A. foetidus-associated host crop species. Likewise, the amounts of ROS-degrading antioxidants (glutathione reductase (GR), peroxidase (POD), ascorbic acid oxidase (AAO), superoxide dismutase (SOD), catalase (CAT)) as well as phenolics were increased, while the amounts of proline and abscisic acid (ABA) were decreased in fungal-associated test crops. Total lipids, proteins and sugars were noted to be high in A. foetidus-associated test crops. From the results, we concluded that A. foetidus have a role in heat stress mitigation that might help to sustain the production of important crops in the future.

The recent work aims at the use of Pantoea conspicua (MT5) and Aspergillus niger (CRS3) to assess their bioremediation potential and growth restoration of Helianthus annuus L. under chromate (Cr+6) stress. The growth of the P. conspicua and A. niger was tested in Cr+6 supplemented media. The strains can withstand up to 1200 and 900 ppm respectively in the media and effectively bio-transform it to nontoxic form. Supplemented metal’s levels significantly decreased the growth attribute of H. annuus (p

Background In the list of abiotic stresses, salt stress is a main growth retarding factor which affects 7% of rain-fed while 30% worldwide irrigated agriculture. However, various strategies are assumed to manage this problem, but the use of endophytes is cheap and eco-friendly. The goal of this study was to evaluate the behavior of endophytic Aspergillus awamori (EWF) in creating salt tolerance in mung bean in terms of its seedling growth, biochemical indices, antioxidant enzymes, endogenous IAA, and ionic status of the plant. Results The results revealed that the 150 mM of NaCl reduced seedling growth (seedlings’ weight and length; leaves number), chlorophyll contents, and IAA. On the other hand, proline, polyphenols, flavonoids, tannin, lipid peroxidation, catalase, and ascorbate peroxidase were increased. Inoculation of EWF had promoted the mung bean growth under all tested conditions. EWF enhanced the biomass and IAA contents of the mung bean plants under salt stress. Moreover, EWF-associated mung bean seedlings exhibited low accumulation of stress markers, and Cl, Na, Na/K, and Ca/K ratio, whereas higher concentrations of Ca, Mg, K, N, and P in mung bean seedlings. Conclusion The results provided a sustainable approach in using endophytic EWF under salt stress, thus concluded that this fungus can be very handy in mung bean as well as other important crop production in saline areas.

The use of plant endophytes to release salt stress is cheap and quick. In this respect, we have isolated an endophytic fungus (Trichoderma reesei) from Solanum surattense that promoted the wheat growth under salt stress. The results showed that the fungal inoculated wheat plants had higher chlorophyll a and b, carotenoids, reduced glutathione (GSH), ascorbate, peroxidase, catalase, weight of plant, flavonoids, total soluble protein, stomatal conductance, transpiration rate, and relative water contents. Also, T. reesei treated plants showed higher IAA, GA, Ca and K, while lower ABA, H2O2, phenol, sugar, proline, electrical conductivity, malondialdehyde (MDA), Na+ and Cl−. The results concluded that the integrative use of T. reesei might help the wheat plants to stand salt stress.

Indole-3-acetic acid (IAA) plays an important role in plant microbe interactions. Current study explored the role of indole-3-acetic acid (IAA) as a signalling molecule for chemical dialogue between endophytic fungus and host plant roots. The endophytic fungus was isolated from the leaves of drought stressed Withenia somnifera and was identified as Aspergillus awamori Wl1 through ITS region of 18 S rDNA sequence. The isolated Wl1 strain was capable of producing important secondary metabolites, including IAA, phenols and sugars. Culture filtrate of the strain contained 24.2, 275.4 and 127.4 μg/mL of IAA, phenols and sugars, respectively. The strain has efficiently colonized the maize roots and enhanced the growth of host plant. In order to determine the role of IAA in root colonization by endophyte, we inhibited the biosynthesis of IAA. Inhibition of IAA production by foliar application of yucasin effectively reduced the colonization of endophyte in maize roots by 52%, whereas root application of yucasin has decreased the colonization frequency by 66%. However, the application of IAA restored the ability of Aspergillus awamori to colonize maize roots and significantly improved various growth parameters. From these observations, it is concluded that IAA plays a vital role in initiating a crosstalk between the two partners.

One of the main problems of the industrialized world is the accumulation of chromium (Cr) in soil, which is a serious threat to the crops. Complete removal of Cr from the contaminated soils poses a great challenge. However, this issue can be minimised by using plant growth promoting microbes as a bioremediation tool. In the present study, healthy plants established near the University campus in Mardan were selected for the isolation of Cr resistant endophytes. From the designated plants, 114 species of endophytic fungi were isolated. Among the 114 isolated strains, 4 strains have induced resistance in L. sativa against Cr. The strains were identified as Aspergillus fumigatus, Rhizopus sp., Penicillium radicum and Fusarium proliferatum based on ITS region (18 S rDNA) homology. The isolates have removed Cr from soil and culture media as well as bio-transformed it from highly toxic hexavalent to least toxic trivalent form, thus helped the Cr stressed L. sativa to restore its normal growth. The Rhizopus Sp. CUC23 has mainly accrued Cr and detoxified intracellularly, whereas A. fumigatus ML43 and P. radicum PL17 has detoxified up to 95% of Cr extracellularly. From the results, it is concluded that the selected endophytic strains might be used as biofertilizer for healthy and safe crop production in Cr contaminated soils.

Endophytes can serve as plant growth promoters as they secret a vast array of phytohormones to support host plants. Keeping the growth promoting activity of the endophytes in view, two endophytic fungi, Asprgillus fumigatus TS1 and Fusarium proliferatum BRL1 have been isolated from the roots of Oxalis corniculata. The isolates have been screened initially for growth promoting activities, including siderophores activity, phosphate solubilization, and secreation of indole acetic acid and gibberellins. Further, the isolates have assayed for the ability to promote the growth of mutant rice Waito-C. The plants associated with TS1 and BRL1 have shown higher chlorophyll content, root-shoot length, and biomass production. The growth promoting activity of the endophytes can be attributed to the various types of GAs and IAA that have been observed in the culture filtrates of the endophytes by the Gas chromatography/mass spectrometry (GC/MS). The GC/MS analysis revealed the presence of different gibberellins concentrations (ng/ml) in TS1 and BRL1 culture filtrate, i.e. GA1 (0.091 ± 0.009, 0.392 ± 0.007), GA3 (0.324 ± 0.077, 0.089 ± 0.0007) and GA7 (0.023 ± 0.002, 0.492 ± 0.005), respectively. Besides, a significant up regulation of plant endogenous GA1 (12.443 ± 0.454 and 15.434 ± 0.245) has been obsereved in TS1 and BRL1 associated plants compared to the control. Moreover, semi quantitative RT-PCR has confirmed the presence/invovment of GA pathways genes (P50–1, P450–3, P450–4, ggs2, and des). The results conclude that the endophytes isolated in this study can ably synthesize bioactive compounds, which play an important role in plant growth promotion.

The antidiabetic and antioxidant potential of ethanolic extract ofCassia nemophilapod (EECNP) was evaluated by three in vitro assays, including yeast glucose uptake assay, glucose adsorption assay, and DPPH radical scavenging activity. The result revealed that the extracts have enhanced the uptake of glucose through the plasma membrane of yeast cells. A linear increase in glucose uptake by yeast cells was noticed with gradual increase in the concentration of the test samples. Moreover, the adsorption capacity of the EECNP was directly proportional to the molar concentration of glucose. Also, the DPPH radical scavenging capacity of the extract was increased to a maximum value of 43.3% at 80 μg/ml, which was then decreased to 41.9% at 100 μg/ml. From the results, it was concluded that EECNP possess good antidiabetic and antioxidant properties as shown by in vitro assays.

The association between microbes and plant roots involves a complex chemical dialogue featured by IAA and flavonoids in addition to other signals. Current study is focused on the role of IAA and flavonoids as essential elements of the chemical dialogue between an endophytic fungus and maize roots. The strain was isolated from the leaves of Withania somnifera and was identified as Aspergillus fumigatus greenish by 18S rDNA sequence. Culture filtrate of the strain contained phyotstimulant (IAA, ammonia and ACC) and signaling compounds including IAA, flavonoids and flavonols. The strain effectively colonized the maize roots and enhanced its growth. In order, to determine the effect of IAA and flavonoids on the ability of the endophyte to colonize maize roots, we inhibited the release of IAA and flavonoids in maize separately which reduced the colonization of endophyte in maize root to 66% and 55% of the untreated control. Similarly, application of IAA in combination with endophyte enhanced root colonization by 90% in zone of cell division, 66% in the zone of elongation and 30% in the zone of maturation. These findings determined that IAA and flavonoids are among the key players of the complex chemical dialogue between plant root and fungal endophyte.

In this study, an endophytic fungus, Aspergillus niger CSR3, was isolated from Cannabis sativa. The culture filtrate (CF) was initially screened for growth-promoting activities such as the presence of siderophores, phosphate solubilization, and the production of indole acetic acid (IAA) and gibberellins and was further assayed for its ability to promote the growth of mutant waito-C rice. Nearly all plant growth attributes examined (root–shoot length, biomass, and chlorophyll content) were significantly enhanced by treatment with CSR3. This growth promotion action was due to the presence of various types of gibberellins (GAs) and IAA in the endophyte CF. Moreover, the presence of GA pathway genes (P50-1, P450-3, P450-4, ggs2, and des) was confirmed by means of semi-quantitative RT-PCR. Finally, the application of CSR3 spore suspension with uniconazole and yucasin on maize seedlings revealed that, similar to exogenous IAA and GA3, CSR3 has the potential to alleviate the inhibitory effect of these inhibitors.

Background Long-term administration of metformin in Type 2 Diabetes mellitus (T2DM), interferes with the absorption of vitamin B12 resulting in deficiency. Low levels of vitamin B12 are associated with cardiovascular morbidity and mortality in T2DM patients. Several genetic variants are associated with vitamin B12 deficiency; one among them is Methylene Tetrahydrofolate reductase (MTHFR). We analyzed MTHFR (rs180133) polymorphism and its association with metformin induced vitamin B12 deficiency in T2DM patients. Methods Three hundred T2DM patients were included. For the study and they were screened for vitamin B12 deficiency markers – Methyl malonic acid (MMA), Homocysteine (Hcy) and high sensitive C reactive protein (hsCRP). Genetic variant was analyzed by ARMS-PCR method. Data was analyzed with various statistical tools like ROC, Odds ratio and Likelihood ratio. Results There is significant reduction in folic acid and vitamin B12 in metformin users. HsCRP, Hcy and MMA are significantly increased with P Conclusion We found an association between MTHFR rs180133, 677C > T and vitamin B12 status in T2DM patients on metformin therapy. Folic acid, MMA and Hcy were found to have high specificity in concordance with MTHFR which showed to be a good predictor for vitamin B12 deficiency in T2DM patients on metformin therapy.

Heavy metal (HM) pollution is of great concern in countries like Pakistan where a huge proportion of human population is exposed to it. These toxic metals are making their way from water bodies to soil where it not only interferes with plant growth and development but also initiates serious health issues in human consuming the produce of such soils. Bioremediation is one of the most viable and efficient solution for the problem. Purpose of the current study was to isolate endophytic fungi from plants grown on HM contaminated soil and screen them for their ability to tolerate multiple HM including chromium (Cr6+), manganese (Mn2+), cobalt (Co2+), copper (Cu2+) and zinc (Zn2+). Out of 27 isolated endophytes, only one strain (MHR-7) was selected for multiple heavy metals tolerance. The strain was identified as Mucor sp. by 18 S and 28 S ribosomal RNA internal transcribed spacer (ITS) 1 and 4 sequence homology. The strain effectively tolerated up to 900 µgmL−1 of these heavy metals showing no remarkable effect on its growth. The adverse effect of the heavy metals, measured as reduction of the fungal growth increased with increasing concentration of the metals. The strain was able to remove 60–87% of heavy metals from broth culture when supplied with 300 µg mL−1 of these metals. A trend of decline in bioremediation potential of the strain was observed with increasing amount of metals. The strain removed metals by biotransformation and/or accumulation of heavy metal in its hyphae. Application of Mucor sp. MHR-7 locked down HM in tis mycelium thereby making them less available to plant root reducing HM uptake and toxicity in mustard. Besides its bioremediation potential, the strain was also able to produce IAA, ACC deaminase and solubilize phosphate making it excellent phytostimulant fungus. It is concluded that MHR-7 is an excellent candidate for use as biofertilizer in fields affected with heavy metals.

Rhizobacteria are known to produce a variety of signal molecules which may modify plant growth by interfering with phytohormone balance. Among the microbial signals are phytohormones, known to contribute to plant endogenous pool of phytohormones. The current chapter describes different methods to study the regulation of gene expression in root apical meristem in response to rhizobacterial inoculation. We describe protocol for the detection of in planta modulation of CKs and IAA by rhizobacteria and their impact on root growth, dissecting the underlying plant signaling pathway by RNA sequencing.

Plants are multicellular organism composed of different types of cells. These all kinds of cells are formed from pluripotent stem cells present at different positions in plant called stem cell niches. All these stem cell niches and their boundaries are maintained by complex regulatory mechanism at molecular level involving different genes, cofactors, and phytohormones. In this chapter, we discussed the regulatory mechanism and models of stem cell maintenance, specifying their boundaries at different stem cell niches.

Besides acting as growth inducing molecule, Gibberellin (GA3) also confers the compatibility of microbial interactions with host. We inoculated 11 days oldZ. maysseedlings grown under hydroponic conditions and high GA3levels withBipolaris sorokiniana(BIPOL) at the spore density (SD) of OD0.6. The high level of GA3negatively affected the growth of the seedlings, accompanied by the high level of stress deducing secondary metabolites (proline, total flavanoids, phenylpropanoids, and glucosinolides). Moreover, high level of GA3produced a hypersensitive response (HR) in the seedlings. The HR developed cross talks with IAA and trans-zeatins and triggered higher production of hypersensitive inducing biomolecules. The other HR co-related biological processes were demonstrated by high phytoalexins level and high protease activities. Such activities ultimately inhibited the colonization of BIPOL on the roots of maize seedlings. The products of the genes expressed at high GA3also conferred the deterrence of BIPOL colonization at SD = OD0.6. Intriguingly, when we inhibited GA3biosynthesis in the seedlings with aerially sprayed uniconizole, prior to BIPOL treatment, the BIPOL colonized and subsequently promoted the seedling growth. This low level of GA3after BIPOL treatment checked the high level of secondary metabolites and hypersensitivity inducing molecules. The results, thus suggested that the aforementioned processes only happened in the BIPOL at SD (OD0.6), whereas the SD at lower levels (OD0.2or OD0.4) neither promoted the growth of uniconizole pre-treated seedlings nor produced HR in control seedlings of maize plant.

Fungal endophytes are known to secrete a large array of secondary metabolites (phenols, flavonoids, indole acetic acid (IAA) etc.) that facilitate crops under stress conditions. Considering this, a potent plant growth promoting endophyte (SXSp1) from the spines and leaves of Solanum xanthocarpum L. has been isolated. The isolated strain ably secreted high quantities of indole-3-acetic acid, phenols and flavonoids. Also, it exhibited phosphate solubilization, siderophore and had 2,2 diphenyl-1-picrylhydrazyl (DPPH) scavenging activity. The SXSp1 also resisted the salinity stress up to 150 mM. LC/MS analysis of SXSp1 culture filtrate (CF) revealed the presence of p-hydroxyl benzoic acid, diadzein, genistien, myricetin and caffeoyl-d-glucose. Moreover, the inoculation of maize plants with SXSp1 significantly (P=0.05) promoted the chlorophyll and carotenoid contents, root and shoot lengths, plant fresh and dry weights, catalase and peroxidase activities, proline, phenolics, flavonoids and relative water contents (RWCs) under salinity. More interestingly, SXSp1-associated plants showed lower endogenous abscisic acid (ABA) and higher endogenous IAA contents that helped the plants to resist salinity stress up to 100 mM. After sequencing, internal transcribed spacer (ITS) regions (ITS1 and ITS4) and phylogenetic analysis, the SXSp1 was identified as Meyerozyma caribbica.

The current climate changes in the form of global warming are one of the leading threats to agricultural crops (including soybean and sunflower). To enable the crops to cope with the heat stress, innovative steps are needed to be taken as soon as possible. Fungal endophytes are known to secrete secondary metabolites that promote the growth of host plants under stress conditions. Therefore, we have isolated endophytic fungus fromEuphorbia indica(a wild desert plant) and tested it for plant growth promoting activities. The culture filtrate of the fungal strains exhibited the presence of secondary metabolites. Higher amounts of indole acetic acid (IAA), salicylic acid (SA), flavonoids, and phenolics have been found in the culture filtrate. The 18S rDNA sequence homology and phylogenetic analysis revealed that the isolate isAspergillus flavus. Soybean and sunflower seedlings were inoculated with the identifiedA. flavus. TheA. flavus-associated seedlings along with the control (withoutA. flavus) were monitored for thermal stress resistance in a growth chamber, operated at 25°C and 40°C. Control seedlings exposed to high temperature stress had higher levels of abscisic acid (ABA), proline, and lower levels of phenols, flavonoids, catalase, and ascorbic acid oxidase. Similarly, a higher reduction in chlorophyll, root-shoot length, and dry weight has been noticed in the control seedlings. The results suggested the usefulness ofA.flavusin host plant growth promotion under heat stress conditions.

Endophytic fungi have been considered as strong plant growth promoters due to phytohormones production. The current study reports the isolation of endophytic fungi from bio-prospective medicinal plant cannabis sativa. Endophytic fungus Bipolaris sp. CSL-1 from the leaves of C. sativa was isolated. Culture filtrate (CF) was primarily investigated for indole-3-acetic acid (IAA) and gibberellins (GAs) and was further evaluated for its capability to enhance mutant Waito-C rice growth attributes. A variety of plant growth characteristics, including seedling length, seedling biomass, and chlorophyll content, were significantly promoted by the CF, and the growth-promoting effect was due to IAA and various GAs in the CF. Gas chromatography/mass spectrometry analysis revealed the quantities (ng/mL) of various GAs, including GA1 (0.758 ± 0.005), GA3 (0.00015 ± 0.005), GA4 (0.945 ± 0.081), GA7 (0.6382 ± 0.012), GA9 (0.0125 ± 0.0002), and GA24 (0.0139 ± 0.0013). Similarly, endogenous GA4 (33.243 ± 4.36), GA24 (29.64 ± 2.68), GA7 (22.5 ± 1.3), and GA12 (25.21 ± 2.8) were significantly upregulated in rice mutant after CF application. Furthermore, RT-PCR indicated that IAA and GA pathway genes (des, ggs2, P50-1, P450-4, and iaaH) were expressed in CSL-1 and the combined application of CSL-1 spore suspension with yucasin and uniconazole to maize seedlings revealed that CSL-1 such as exogenous GA3 and IAA alleviated the negative effect of uniconazole and yucasin and promoted maize-seedling growth. These findings suggest that the endophytic fungus CSL-1, which produces IAA and GAs, can play a vital role in promoting plant growth and that CSL-1 could be used to enhance crop growth and mitigate plant stress under a variety of environmental conditions.

Urinary tract infection (UTI) is the most common nosocomial infection among hospitalized patients. Area-specific monitoring studies aimed to gain knowledge about the type of pathogens responsible for UTIs and resistance pattern of the causative agents may help clinicians to choose correct treatment regimen. So, the present study was aimed to investigate the antibiotic resistance of urinary pathogens isolated at Chittagong Medical College Hospital (CMCH) during January to June, 2007. Out of 360 clinical samples of urine collected, 175 (48.61%) showed significant bacterial growth. The most common pathogens isolated were Escherichia coli (66, 37.71%), Klebsiella species (60, 34.29%), Proteus species (17, 9.71%) and Pseudomonas species (16, 9.14%). Members of the Enterobacteriaceae were 80%-100% sensitive to Imipenem while they were found variably sensitive to other commonly used antibiotics. Pseudomonas species were 100% sensitive to Imipenem, 75% to Amikacin, and 50% to Ceftazidime. Staphylococcus aureus was found 100% sensitive to Vancomycin, 80% to Gentamicin and Amikacin each, and 55% to Oxacillin. Enterococcus species were 80% sensitive to Imipenem, 60% to Ciprofloxacin, 50% to Vancomycin and Amikacin each. The clinicians should use Imipenem selectively in cases of un-responsiveness to commonly used antibiotics. Bangladesh J Med Microbiol 2008; 02 (02): 17-21

Background To combat salinity, plants need easily accessible, safe and sustainable mechanisms for optimum growth. Recently, endophytes proved to be the promising candidates that helped the host plant to thrive under stress conditions. Therefore, the aim was to discover endophytic strain(s) and their mechanism of action to alleviate salt stress in maize. Results Keeping the diverse role of endophytes in view, 9 endophytic fungi from the spines of Euphorbia milli L. were isolated. Among the isolated fungal isolates, isolate FH1 was selected for further study on the basis of high antioxidant activity and capability to produce high indole-3-acetic acid (IAA), indole-3-acetamide (IAM), phenol and flavonoid contents. The 18S rDNA sequence homology and phylogenetic analysis of the fungal isolate FH1 revealed to be Yarrowia lipolytica. Furthermore, the inoculation of Y. lipolytica FH1 had significantly promoted plant growth attributes in treated maize as compared to positive (salt stress) and negative (salt stress free) controls. Likewise, differences in chlorophyll, carotenes, electrolyte leakage, leaf relative water, peroxidase, catalase, ABA, IAA and proline contents were observed between treated maize and controls. Interestingly, Y. lipolytica FH1 inoculated plants showed lower endogenous ABA and higher endogenous IAA contents. Conclusion From the results, we have concluded that Y. lipolytica inoculation has promoted the growth of maize plants through controlled metabolism and hormonal secretions (ABA and IAA) under salinity stress. Because of the fact, Y. lipolytica can be tried as an eco-friendly bio-fertilizer to achieve optimum crop productivity under saline conditions.

Cinnamic acid (CA) is an allelochemical that inhibits the growth of root promoting soil microorganisms. To prevent the growth of soil microbes, CA modulates several metabolic pathways in host plants and soil microbes. The aim of the current study was to investigate the effect of CA on maize root growth, exudation of secondary metabolites and its interaction with beneficial endophyte Pz11. The endophyte Pz11 was isolated from the roots of drought stressed Asphodelus tenuifolius (wild onion). The Pz11 strain was identified as Fusarium culmorum by homology of the internal transcribed spacer (ITS) region of 18 S rDNA sequence. The F. culmorum Pz11 produced phytostimulants and signaling compounds, such as indole-3-acetic acid (IAA), flavonoids and sugars. Moreover, the strain have effectively colonized the roots of maize and subsequently enhanced the growth of its host plants. On the contrary, application of CA has reduced root growth in maize seedlings as well as root colonization ability of F. culmorum Pz11. Also, maize seedlings exposed to CA exude low quantities of flavonoids and polyphenols. In conclusion, CA reduces the maize root growth and exudation of secondary metabolites, which may affects its ability to attract plant growth promoting endophytic fungi.

Background Medicinal plants have been founded as traditional herbal medicine worldwide. Most of the plant’s therapeutic properties are due to the presence of secondary metabolites such as alkaloids, glycosides, tannins and volatile oil. Methods The present investigation analyzed the High-Pressure Liquid Chromatography (HPLC) fractions of Glycyrrhiza glabra (Aqueous, Chloroform, Ethanol and Hexane) against multidrug resistant human bacterial pathogens (Escherichia coli, Acinetobacter baumannii, Staphylococcus aureus and Pseudomonas aeruginosa). All the fractions showed antibacterial activity, were subjected to LC MS/MS analysis for identification of bioactive compounds. Results Among total HPLC fractions of G. glabra (n = 20), three HPLC fractions showed potential activity against multidrug resistant (MDR) bacterial isolates. Fraction 1 (F1) of aqueous extracts, showed activity against A. baumannii (15 ± 0.5 mm). F4 from hexane extract of G. glabra showed activity against S. aureus (10 ± 0.2 mm). However, F2 from ethanol extract exhibited activity against S. aureus (10 ± 0.3 mm). These active fractions were further processed by LC MS/MS analysis for the identification of compounds. Ellagic acid was identified in the F1 of aqueous extract while 6-aldehydo-isoophiopogonone was present in F4 of hexane extract. Similarly, Liquirtigenin was identified in F2 of ethanol. Conclusions Glycyrrhiza glabra extracts HPLC fractions showed anti-MDR activity. Three bioactive compounds were identified in the study. 6-aldehydo-isoophiopogonone and Liquirtigenin were for the first time reported in G. glabra. Further characterization of the identified compounds will be helpful for possible therapeutic uses against infectious diseases caused by multidrug resistant bacteria.

We have isolated an endophytic fungus with heat stress alleviation potential from wild plant Euphorbia indica L. The phylogenetic analysis and 18S rDNA sequence homology revealed that the designated isolate was Aspergillus japonicus EuR-26. Analysis of A. japonicus culture filtrate displayed higher concentrations of salicylic acid (SA), indoleacetic acid (IAA), flavonoids, and phenolics. Furthermore, A. japonicus association with soybean and sunflower had improved plant biomass and other growth features under high temperature stress (40°C) in comparison to endophyte-free plants. In fact, endophytic association mitigated heat stress by negotiating the activities of abscisic acid, catalase, and ascorbic acid oxidase in both soybean and sunflower. The nutritional quality (phenolic, flavonoids, soluble sugars, proteins, and lipids) of the A. japonicus-associated seedlings has also improved under heat stress in comparison to endophyte-free plants. From the results, it is concluded that A. japonicus can modulate host plants growth under heat stress and can be used as thermal stress alleviator in arid and semiarid regions of the globe (where mean summer temperature exceeds 40°C) to sustain agriculture.

Indole-3-acetic acid (IAA) is known for their role in plant root interactions with microbial partners. Current study was focused on role of IAA as signal for colonization between endopytic fungus Fusariun oxysporum wlw and maize roots. Culture filtrate of te strain contained 31 of IAA μg/mL of IAA. Addition of tryptophan concentration ranging from 500 to 1000 μg/mL in culture medium significantly enhanced production of IAA by Fusariun oxysporum wlw. The strain effectively colonized the roots of maize and subsequently enhanced the growth and proliferation of host plant. In order to demined the role of IAA in root colonization by F. oxysporum we inhibited the biosynthesis of IAA by using IAA biosynthesis inhibitor wice efficiently reduced te colonization of F. oxysporum in maize roots by 46% (foliar application) and 62% (root application) of the seedlings without yucasin (IAA biosynthesis inhibitor) treatment suggesting an IAA crosstalk between the two partners. Exogenous application of IAA restored the ability of endophyte F. oxysporum to colonize maize roots and significantly improved different growth parameters of maize seedlings. It is concluded that a molecular crosstalk of maize roots and endophytic F. oxysporum wlw is necessary for subsequent endophytic association between them.

Plant growth and gibberellins (GAs) biosynthesis are two separate but linked processes, involving many genes but fewer have been reported for their role in plant growth and development. Due to little information on the genes involved in such processes, the known plant growth promoting and GAs producing fungal endophyte Penicillium citrinum KACC43900 was used as potential tool to obtain a blueprint of the putative growth promoting and GAs synthesizing proteins. For proteomic analysis, the seedlings of Glehnia littoralis were treated with culture filtrate of P. citrinum KACC43900, which revealed significant differences between 2 dimensional gel electrophoresis profile of the crude protein extracts of the treated and control samples. Matrix-assisted laser desorption/ionization analysis of the 56 selected spots led to the identification of 41 proteins. A significant number (31.5%) of these highly expressed proteins were associated with plant growth regulation, including beta-expansin EXPB4, ent-kaur 16-ene synthase, gibberellin 3-oxidase, and cytochrome P450 family proteins. Proteins involved in regulating energy metabolism and intracellular redox conditions, such as glyceraldehyde-3-phosphate dehydrogenase and ribulose-1,5-bisphospahate carboxylase/oxygenase, were also expressed. It was concluded that culture filtrate of P. citrinum KACC43900 altered the gene expression pattern of host G. littoralis. Current study highlighted the importance of proteomics as a starting tool for any post-genomic research.

Ethnomedicinal knowledge of plant-derived bioactives could help us in discovering new therapeutic compounds of great potential. Certainly, dandelion has been used in traditional ethno-medicinal systems (i.e., Chinese, Arabian, Indian, and Native American) to treat different types of cancer. Though, dandelion is highly vigorous, but the potential mode of action is still unclear. In the current study, the antiproliferative activity of methanolic extracts of dandelion root (MEDr) on cell viability of HepG2, MCF7, HCT116, and normal Hs27 was investigated. It was observed that MEDr (500 μg/mL) drastically decreased the growth of HepG2 cell line, while the effect on MCF7 and HCT116 cell lines was less pronounced and no effect has been observed in Hs27 cell lines. The MEDr also enhanced the phosphorylation level of AMPK of HepG2 cells, which considered crucial in cancer treatment and other metabolic diseases. The AMPK activation by MEDr noticed in the current study has never been reported previously. The results regarding the number of apoptotic cells (HepG2 cells) were in line with the cell viability test. The current observations clearly demonstrated the potency of MEDr against liver cancer with validation that dandelion could control AMPK and thus cancer in the treated cell lines.

Abiotic stress resistance strategies are powerful approaches to sustainable agriculture because they reduce chemical input and enhance plant productivity. In current study, an endophytic fungus, Aspergillus flavus CHS1 was isolated from Chenopodium album Roots. CHS1 was initially screened for growth promoting activities like siderphore, phosphate solubilization, and the production of indole acetic acid and gibberellins and were further assayed for its ability to promote the growth of mutant Waito-C rice. The results revealed that different plant growth characteristic such as chlorophyll content, root-shoot length, and biomass production were significantly promoted during CHS1 treatment. This growth promotion action was due to the presence of various types of GAs and IAA in the endophyte culture filtrate. Significant up regulation with respect to levels in the control was observed in all endogenous plant GAs, after treatment with CHS1. Furthermore, to evaluate the potential of CHS1 against NaCl stress up to 400 mM, it was tested for its ability to improve soybean plant growth under NaCl stress. In endophyte-soybean interaction, CHS1 association significantly increased plant growth and attenuated the NaCl stress by down regulating ABA and JA synthesis. Similarly, it significantly elevated antioxidant activities of enzymes catalase, polyphenoloxidase, superoxide dismutase and peroxidase as compared to non-inoculated salt stress plants. Thus, CHS1 ameliorated the adverse effect of high NaCl stress and rescued soybean plant growth by regulating the endogenous plant hormones and antioxidative system. We conclude that CHS1 isolate could be exploited to increase salt resistant and yield in crop plants.

Lotus japonicus possesses major advantages as a model legume for the study of plant-microbe interactions. The relative absence of genetic information on its normal microbial partner (i.e., Mesorhizobium loti) could limit its utility in research. Here we show for the first time that the broad host range Rhizobium strain NGR234 nodulates and fixes nitrogen in symbiosis with Lotus japonicus ecotypes "Gifu" and "Funakura". We demonstrate that bacterial mutants deficient in nodulation or nitrogen fixation possess the expected phenotype with L. japonicus. Nodulation of L. japonicus was sensitive to nitrate. Vermiculite was an efficient synthetic growth substrate, allowing axenic growth in Magenta jars. The genetic analysis of the Lotus japonicus-Mesorhizobium interaction should be accelerated through the use of this well-defined microsymbiont.

In the pursuit of sustainable agriculture through environment and human health friendly practices, we evaluated the potential of a novel gibberellins (GAs) producing basidiomycetous endophytic fungus Porostereum spadiceum AGH786, for alleviating salt stress and promoting health benefits of soybean. Soybean seedlings exposed to different levels of NaCl stress (70 and 140 mM) under greenhouse conditions, were inoculated with the AGH786 strain. Levels of phytohormones including GAs, JA and ABA, and isoflavones were compared in control and the inoculated seedlings to understand the mechanism through which the stress is alleviated. Gibberellins producing endophytic fungi have been vital for promoting plant growth under normal and stress conditions. We report P. spadiceum AGH786 as the ever first GAs producing basidiomycetous fungus capable of producing six types of GAs. In comparison to the so for most efficient GAs producing Gibberella fujikuroi, AGH786 produced significantly higher amount of the bioactive GA3. Salt-stressed phenotype of soybean seedlings was characterized by low content of GAs and high amount of ABA and JA with reduced shoot length, biomass, leaf area, chlorophyll contents, and rate of photosynthesis. Mitigation of salt stress by AGH786 was always accompanied by high GAs, and low ABA and JA, suggesting that this endophytic fungus reduces the effect of salinity by modulating endogenous phytohormones of the seedlings. Additionally, this strain also enhanced the endogenous level of two isoflavones including daidzen and genistein in soybean seedlings under normal as well as salt stress conditions as compared to their respective controls. P. spadiceum AGH786 boosted the NaCl stress tolerance and growth in soybean, by modulating seedlings endogenous phytohormones and isoflavones suggesting a valuable contribution of this potent fungal biofertilizer in sustainable agriculture in salt affected soils.

The NAC domain genes encode a large family of plant-specific transcription factors that play diverse roles in plant development and stress regulation. In this study, a total of 60 full-length putative NAC genes were isolated from Gossypium hirsutum L. Based on their phylogeny, all GhNAC genes were clustered into seven distinct subfamilies, which exhibit functional similarity. Similarly, a phylogenetic tree for GhNAC genes and their motif showed close resemblance among the subfamilies. The isolated 60 full-length GhNAC genes were located on 13 different chromosomes of D sub-genome. Majority of the NACs showed specific temporal and spatial expression patterns in tissue-specific (fibers, cotyledon leaf, mature leaf, stem, root and flower) studies based on qRT-PCR analyses. Furthermore, the roles of GhNAC genes were monitored using qRT-PCR during leaf senescence and following treatment with ethylene, abscisic acid, gibberellic acid and drought or salinity. This first comprehensive study of GhNAC family elucidates the essential role of these genes in cotton development and in response to various stresses. This study lays fundamental foundations for future research and development in cotton genome.

Sun drying of apricot fruit (Prunus armeniaca L.) is the oldest form of preservation technique known to the people of Ladakh and is still practiced on a large scale. This is because of the abundance of available sunshine. Export of fresh fruits of apricot and apple outside the region is restricted through quarantine regulations to check the entry of a noxious pest, codling moth to other temperate fruit belts (J&K and HP) of the country. Henceforth, drying is the only possible venture in vogue to receive income from apricot cultivation. Traditional practices followed commonly render the final produce to be unhygienic thereby fetching low returns. The introduction of different types of solar dryers along with other novel drying processes has improved this age-old preservation technique. A perusal of various drying practices followed and the advantages cum utilisation pattern of various driers are discussed in the paper.

Objective Biofilms are complex, multi-species bacterial communities that colonize the oral cavity in the form of plaque and are known to cause dental caries and periodontal disease. Present study demonstrated the potential of three selected medicinal plants against isolated and identified dental biofilm forming strains. Methods Pathogenic bacteria from dental biofilms were isolated, cultured, identified by phylogenetic analysis using PCR-based 16S ribosomal RNA (or 16S rRNA) nucleotide sequences and were analyzed for their biofilm forming capability. The antimicrobial activity of the three important medicinal plant extracts (Acacia arabica, Tamarix aphylla L. and Melia azadirachta L.) was determined against the highest biofilm forming bacteria. Results Phylogenetic analysis revealed that the 19 strains belonged to Proteobacteria, Firmicutes and Actinobacteria. Among the 19 isolates, eleven strains were found to possess high biofilm formation capability comparatively and antimicrobial activity assay showed that the selected plants considerably inhibited their growth. Extract from A. arabica stem had strong effect on the ability of bacteria isolated from dental carries as evident by up to 73% reduction in biofilm formation on surface pre-treated with extract from this plant. The same extract also showed tremendous biofilm cleaning potential up to 87% of the biofilm. Conclusion The results suggested that the extracts of selected medicinal plants could be used for protection against pathogenic dental biofilm causing bacteria and attempts should be taken by pharmaceutical industries to utilize it in dental caring products.

Microbial associations with plants are crucial for the survival of both the partners. Beside other ways of establishing such associations, phytohormones enjoy a key role in plant-microbe interactions from initial dialog between the two to the establishment of a viable partnership. Cytokinins (CKs) and IAA are among the five classical groups of phytohormones implicated in plant immune response, early signaling, and deciding the fate of interactions between plant and microbes. Here we describe a method to study modulation of Cks and IAA in plant under the influence of a pathogenic bacterium, Pseudomonas syringae tomato DC3000. A method for inoculating bacteria on host plant and subsequent determination of Cks and IAA through HPLC-ESI-MS/MS is described.

Green vegetable crops irrigated with wastewater are highly contaminated with heavy metals and are the main source of human exposure to the contaminants. In this study accumulation of eight heavy metals (Cu, Ni, Zn, Cr, Fe, Mn, Co and Pb) in green vegetables like Allium cepa, Allium sativum, Solanum lycopersicum and Solanum melongena, irrigated with wastewater in Mardan are studied using Atomic Absorption spectrophotometer. The studied metals in vegetable grown on wastewater irrigated soil were significantly higher than those of tube well water irrigated soil and WHO/FAO permissible limits (P

Phytohormones signal and combine to maintain the physiological equilibrium in the plant. Pathogens enhance host susceptibility by modulating the hormonal balance of the plant cell. Unlike other plant hormones, the detailed role of cytokinin in plant immunity remains to be fully elucidated. Here, extensive data mining, including of pathogenicity factors, host regulatory proteins, enzymes of hormone biosynthesis, and signaling components, established an integrated signaling network of 105 nodes and 163 edges. Dynamic modeling and system analysis identified multiple cytokinin-mediated regulatory interactions in plant disease networks. This includes specific synergism between cytokinin and salicylic acid pathways and previously undiscovered aspects of antagonism between cytokinin and auxin in plant immunity. Predicted interactions and hormonal effects on plant immunity are confirmed in subsequent experiments with Pseudomonas syringae pv tomato DC3000 and Arabidopsis thaliana. Our dynamic simulation is instrumental in predicting system effects of individual components in complex hormone disease networks and synergism or antagonism between pathways.

Efficient callus and explant regeneration medium, using microbial extract (SPE purified) or supernatant has been formulated for Brassica oleracea L. var. capitata. Two cyanobacterial strains (Anabaena sp. Ck1 and Chroococcidiopsis sp. Ck4) and two bacterial strains, (Pseudomonas spp. Am3 and Am4) known to produce a number of cytokinins, tZ, cZ, ZR, DHZR and IAA were selected for the media formulation. Supernatant from strains with high cytokinin to IAA ratio, including Pseudomonas aeruginosa Am3 (2.08) and Chroococcidiopsis sp. Ck4 (0.8) efficiently induced compact calli which were turned green upon exposure to light. The strains producing lower cytokinins to IAA ratio (0.11-0.13) on the other hand induced friable callus which were unable to regenerate on the selected media combinations. Leaf, stem and root explants of Brassica oleracea L. regenerated on MS medium supplemented with phytohormones from microbial origin with efficiency comparable to standard cytokinins and IAA. Supplements from cyanobacterial origin proved to be the best for induction of adventitious roots and shoots on internodal and petiolar segments. Hypocotyl explants however, responded well on MS supplemented with bacterial metabolites. Induction of adventitious shoots on root explants was supported by phytohormones from both origin equally well. Callus induction on the seeds and regeneration of shoots on calli was also observed. Cyanobacteria based media were more efficient to induce calli capable of regeneration upon exposure to light. Internodal explants were highly amenable to regenerate shoot and roots simultaneously. Root explants were the less successful to regenerate shoots.

Phytohormones from rhizobacterial origin have been linked to their phytostimulation potential. However, while studying the efficacy of plant growth promoting bacteria, focus has always been on a single hormone. The role of plant hormones often overlay and they mutually modulate their effect. In current study focus was on the role of two hormones (cytokinins and indole acetic acid) in phytostimulation by rhizobacteria. Endogenous rhizosphere bacteria were isolated and screened for the presence of phytohormones. Bacterial strains from three different genera (Pseudomonas, Bacillus and Azospirillum) were screened positive for cytokinins and IAA. Phytohormones were simultaneously determined in SPE purified bacterial extract by ultra performance liquid chromatography (UPLC) coupled to a tandem mass spectrometer through electrospray interface. Cytokinins and IAA were determined in positive and negative mode, respectively with MRM scan. Zeatin, zeatin riboside and dihydrozeatin riboside were detected and quantified in the selected strains. Significant positive correlation between cytokinins and IAA in bacterial culture and plant endogenous hormones (r = 0.933 and r = 0.983; P = 0.01, respectively) was observed. However, strains with high IAA to cytokinins ratio could hardly enhance in-planta cytokinins, indicating antagonistic relation between the two hormones. Significant correlation of cytokinin with shoot length (r = 0.797; P = 0.01), fresh weight (r = 0.685; P = 0.01) and dry weight (r = 0.704; P = 0.01) was reported under axenic conditions. Bacterial IAA was correlated negatively to root length (r = 0.853; P = 0.01) and positively correlated to the number of roots (r = 0.964; P = 0.01). In natural conditions maximum increase in spike length (33%), number of tillers (71%) and weight of seeds (39%) was documented at final harvest in bacterially inoculated plants.