Highest Indole Acetic Acid Production Research Articles

Co‐inoculation with Rhizobium, Azospirillum, and microalgae increases common bean yield and profitability

AbstractWidely used as inoculants in agriculture, nitrogen‐fixing bacteria can be associated with other plant growth‐promoting microorganisms (PGPMs) to increase crop grain yield. This study aimed to evaluate whether co‐inoculation with Rhizobium tropici and Azospirillum brasilense associated with microalgae or cyanobacteria enhances common bean (Phaseolus vulgaris) grain yield and its economic gains in contrasting sandy and clayey soils. Water‐soluble protein content and indole‐3‐acetic acid (IAA) production of two cyanobacteria (Anabaena cylindrica and Calotrix brevissima) and six Chlorophyta microalgae (Nannochloropsis oculata, Haematococcus pluvialis, Muriellopsis sphaerica, Chlorella protothecoides, Chlorella vulgaris, and Botryococcus braunii) were determined. Anabaena cylindrica and C. brevissima had the highest IAA production, 336.7 ± 44.7 and 94.1 ± 11.8 µg IAA mg−1 of protein, respectively. Two field experiments were carried out to evaluate the triple inoculation of these Chlorophyta microalgae and cyanobacteria associated with R. tropici plus A. brasilense on the agronomic efficiency and profitability of common bean. Co‐inoculation of C. vulgaris plus R. tropici and A. brasilense in common bean had grain yield of 1277.5 kg ha−1 in clayey soil and 2960.3 kg ha−1 in sandy soil, increasing by 219.7 and 656.0 kg ha−1, respectively, in relation to double inoculation of R. tropici + A. brasilense. In the sandy soil, common bean with triple co‐inoculation had the highest profit ($956 ha−1), which was 25.6% higher than the N fertilized plants. Co‐inoculation with PGPMs generated the highest economic gains, and in addition, it is an eco‐friendly agronomic practice for sustainable food production.

Elimination of ineffective inorganic salt component in medium for indole‐3‐acetic acid synthesis by Serratia plymuthica UBCF_13 and its effect on the growth of chili seedlings

Indole‐3‐acetic acid (IAA) is an essential phytohormone that controls a variety of plant growth mechanisms. Bacteria can produce IAA to stimulate plant growth, with its production influenced by the culture conditions. Serratia plymuthica UBCF_13 is recognized as an IAA‐producing bacterium, exhibiting maximum IAA production in a yeast medium comprising yeast extract, sucrose, K2HPO4, MgSO4, NaCl, and CaCO3. However, prior studies optimizing individual inorganic salt components indicated minimal impact on IAA synthesis within this medium. This study aimed to eliminate the unnecessary inorganic salt components and the medium was then applied to investigate the IAA biosynthesis pathway and the plant growth‐promoting assay. The elimination assay consisted of yeast sucrose medium devoid of K2HPO4, MgSO4, NaCl, or CaCO3, and yeast sucrose medium containing only MgSO4 and CaCO3. Various indole compounds were then added to the revised medium composition to investigate the IAA biosynthesis pathway of UBCF_13 using high‐performance liquid chromatography (HPLC). Furthermore, the effect of UBCF_13 culture supernatant, cultivated in the new medium, on chili plant growth was evaluated. The highest IAA production (138.8 µg/mL) was observed in the yeast sucrose with CaCO3 and MgSO4 (elimination of K2HPO4 and NaCl). The presence of indole‐3‐acetamide (IAM) compound from the medium extracts, supplemented with multiple indole compounds, revealed that UBCF_13 may use the IAM pathway. The application of UBCF_13 supernatant enhanced the shoot, root length, fresh weight, and germination time of chili seeds by 37.7%, 49.3%, 204.3%, and 38.6%, respectively. This study demonstrated that eliminating K2HPO4 and NaCl provided a new culture medium composition conducive to IAA production by UBCF_13. Moreover, the UBCF_13 extract has the potential to promote plant growth.

Comparative study between Salkowski reagent and chromatographic method for auxins quantification from bacterial production.

The Salkowski reagent method is a colorimetric technique used to determine auxin production, specifically as indole-3-acetic acid (IAA). It was developed to determine indoles rapidly; however, it does not follow Beer's law at high concentrations of IAA. Thus, there could be an overestimation of IAA with the Salkowski technique due to the detection of other indole compounds. This study aims to compare the Salkowski colorimetric method versus a chromatographic method to evidence the imprecision or overestimation obtained when auxins, such as indole-acetic acid (IAA), are determined as traits from promoting growth plant bacteria (PGPB), using ten different strains from three different isolation sources. The analysis used the same bacterial culture to compare the Salkowski colorimetric and chromatographic results. Each bacterium was cultivated in the modified TSA without or with tryptophan for 96 h. The same supernatant culture was used in both methods: Salkowski reagent and ultra-performance liquid chromatography coupled with a Mass Spectrometer (LC-MS/MS). The first method indicated 5.4 to 27.4 mg L-1 without tryptophan in ten evaluated strains. When tryptophan was used as an inductor of auxin production, an increase was observed with an interval from 4.4 to 160 mg L-1. The principal auxin produced by all strains was IAA from that evaluated by the LC-MS/MS method, with significantly higher concentration with tryptophan addition than without. Strains belonging to the Kocuria genus were highlighted by high IAA production. The indole-3-propionic acid (IPA) was detected in all the bacterial cultures without tryptophan and only in K. turfanensis As05 with tryptophan, while it was not detected in other strains. In addition, indole-3-butyric acid (IBA) was detected at trace levels (13-16 µg L-1). The Salkowski reagent overestimates the IAA concentration with an interval of 41-1042 folds without tryptophan and 7-16330 folds with tryptophan as inductor. In future works, it will be necessary to determine IAA or other auxins using more suitable sensitive techniques and methodologies.

Indole-3-Acetic Acid Producing Yeasts in the Phyllosphere of Legumes: Benefits for Chili Growth

Plant growth-promoting yeast (PGPY), which is associated with plants, has demonstrated the ability to enhance plant growth and crop yield and provide an effective alternative strategy for reducing the extensive use of chemical fertilizers. However, plant development and yield effects of the phyllosphere yeast have received comparatively little research. Therefore, 95 phylloplane yeast were isolated from the leaves of plants in Leguminosae, and their capability to produce indole-3-acetic acid (IAA) in yeast extract peptone dextrose (YPD) broth supplemented with L-tryptophan was evaluated in this research. Forty-two isolates were selected, due to their ability to produce IAA. Among these isolates, 7 showed high IAA production of more than 40 mg/g DW in YPD with tryptophan as the precursor. These strains were identified as Candida tropicalis, Pichia kudriavzevii and Tortispora caseinolytica based on morphological and D1/D2 domain of LSU rDNA sequence analysis. Other plant growth promotion traits, including the solubilization of phosphate, production of ammonia and siderophores and ACC deaminase activity, were also investigated. The results revealed that Candida tropicalis KPS2219 exhibited maximum IAA production of 54.10 mg/g DW, high ammonia production at 1.16 mg/mL and siderophore production. In a greenhouse experiment, the ability of C. tropicalis KPS2219 to enhance the growth of chili seedlings was investigated. The results revealed that seed priming followed by foliar spraying with C. tropicalis KPS2219 significantly increased the root length, shoot length, root dry weight and stem dry weight of the seedlings by 17.72, 29.15, 60 and 46.15 %, respectively, in comparison to those of the uninoculated control. These findings indicate the possibility of C. tropicalis KPS2219 as a bioinoculant to promote plant development and the effectiveness of foliar application. The efficacy of employing yeast consortiums to enhance growth will be investigated for further study. HIGHLIGHTS Phylloplane yeasts with remarkably high IAA production and multiple traits that promote the growth of plants were isolated from the new habitat, legumes. Spraying the yeast cells on the leaves of chili plants accelerated the growth of the plant. GRAPHICAL ABSTRACT

Potential of Bacillus sp. from Kebun Raya Liwa as a Producer of Indole Acetic Acid (IAA) Hormone

Kebun Raya Liwa’s soil has the potential to allow for a population of microorganisms such as the Bacillus sp. Bacillus sp. Kebun Raya Liwa are known to produce phytohormones that have the potential to help plant growth and development, one of which is the hormone Indole Acetic Acid (IAA). This study aims to determine the potential of Bacillus sp. from the soil of Kebun Rawa Liwa in producing IAA hormones. The research method included taking soil samples, preparing Nutrient Agar (NA) medium, measuring soil pH samples, isolating soil bacteria, purifying and characterizing Bacillus sp., preparing IAA standard curves, and testing the potency of Bacillus sp. IAA production was carried out in two ways, namely measuring the content of IAA with the addition of tryptophan and without the addition of tryptophan. IAA concentration was measured using a spectrophotometer with a wavelength of 520 nm. The data obtained is presented in tabular form. The isolation results obtained 10 isolates that were able to produce IAA hormone with and without tryptophan. The highest IAA production without tryptophan was isolate BP 5 with a concentration of 1.18 ppm, and the isolate that was able to produce the highest IAA with tryptophan was isolate BP 13 with a concentration of 3.90 ppm. This indicates that the isolate Bacillus sp. from the soil rhizosphere Kebun Raya Liwa can produce IAA hormones.

Development of Halotolerant Microbial Consortia for Salt Stress Mitigation and Sustainable Tomato Production in Sodic Soils: An Enzyme Mechanism Approach

Salt stress caused by sodic soils is an important constraint that impacts the production of crucial solanaceous vegetable crops globally. Halotolerant poly-extremophiles rhizobacteria can inhabit hostile environments like salinity, drought, etc. The present study was aimed to design a halotolerant micro-formulation using highly salt-tolerant bacterial strains previously isolated from salt-tolerant rice and wheat rhizosphere in sodic soil. Nine halotolerant isolates were examined for plant growth-promoting traits and biomass production in pot studies with sodic soil of pH 9.23 in tomato. Compatible, efficient isolates were aimed to be formulated into different consortia like PGPR-C1, PGPR-C2 and, PGPR-C3 for field evaluation in sodic soils of pH 9.14. Halotolerant rhizobacterial consortia (PGPR-C3) comprising Lysinibacillus spp. and Bacillus spp. were found to produce extracellular enzymes like amylase, protease, cellulase, and lipase, showing significantly enhanced vegetative parameters, yield and lycopene content of tomato hybrid NS585 under salt-stressed sodic soils. PGPR-C3 consortia also showed enhanced plant growth-promoting activities and halo tolerance like high Indole acetic acid production, 1-aminocyclopropane-1-carboxylic acid deaminase, and antioxidative enzyme activity over the uninoculated control. Further, inoculation with PGPR-C3 consortia resulted in the efficient exclusion of Na+ ions from the rhizosphere through increased absorption of K+. Results of the study reveal that inoculation with PGPR-C3 consortia could alleviate the salt stress and promotes the successful cultivation of tomato crop in sodic soils. It can be considered the best option for eco-friendly, sustainable cultivation of vegetables like a tomato in sodic soils with a high pH range of up to 9.14.

Waste valorization as low-cost media engineering for auxin production from the newly isolated Streptomyces rubrogriseus AW22: Model development

Indole-3-acetic acid (IAA) represents a crucial phytohormone regulating specific tropic responses in plants and functions as a chemical signal between plant hosts and their symbionts. The Actinobacteria strain of AW22 with high IAA production ability was isolated in Algeria for the first time and was characterized as Streptomyces rubrogriseus through chemotaxonomic analysis and 16 S rDNA sequence alignment. The suitable medium for a maximum IAA yield was engineered in vitro and in silico using machine learning-assisted modeling. The primary low-cost feedstocks comprised various concentrations of spent coffee grounds (SCGs) and carob bean grounds (CBGs) extracts. Further, we combined the Box-Behnken design from response surface methodology (BBD-RSM) with artificial neural networks (ANNs) coupled with the genetic algorithm (GA). The critical process parameters screened via Plackett-Burman design (PBD) served as BBD and ANN-GA inputs, with IAA yield as the output variable. Analysis of the putative IAA using thin-layer chromatography (TLC) and (HPLC) revealed Rf values equal to 0.69 and a retention time of 3.711 min, equivalent to the authentic IAA. AW 22 achieved a maximum IAA yield of 188.290 ± 0.38 μg/mL using the process parameters generated by the ANN-GA model, consisting of L-Trp, 0.6%; SCG, 30%; T°, 25.8 °C; and pH 9, after eight days of incubation. An R2 of 99.98%, adding to an MSE of 1.86 × 10−5 at 129 epochs, postulated higher reliability of ANN-GA-approach in predicting responses, compared with BBD-RSM modeling exhibiting an R2 of 76.28%. The validation experiments resulted in a 4.55-fold and 4.46-fold increase in IAA secretion, corresponding to ANN-GA and BBD-RSM models, respectively, confirming the validity of both models.

Diversity of Endophytic Yeasts from Agricultural Fruits Positive for Phytohormone IAA Production.

This study reports the diversity of cultivable endophytic yeasts from agricultural fruits that respond positively to the plant-promoting property of indole-3-acetic acid (IAA) production. The IAA synthesis by the strains was quantified with an Agilent 1100 series liquid chromatography system. IAA was present in the culture liquid of 72% of all 97 strains examined after three days of cultivation. The most active endophytic yeast strains in this study belonged to the species Aureobasidium pullulans, Candida zeylanoides, Hanseniaspora uvarum, Metschnikowia pulcherrima, Meyerozyma caribbica, Rhodotorula mucilaginosa, and Yarrowia galli. The highest IAA production was observed in the endophytic strain of A. pullulans (9109.19 ± 146.02 μg/g). No significant differences were found between IAA production in strains from agricultural products of different countries. However, the level of IAA production was strictly strain-specific. Our results suggest that the internal tissues of fruits may be a promising source for the isolation of plant-beneficial yeasts that can be used to promote plant growth.

In vitro Screening of Sunflower Associated Endophytic Bacteria With Plant Growth-Promoting Traits

Harnessing endophytic microbes as bioinoculants promises to solve agricultural problems and improve crop yield. Out of fifty endophytic bacteria of sunflowers, 20 were selected based on plant growth-promoting. These plant growth-promoting bacteria were identified as Bacillus, Pseudomonas, and Stenotrophomonas. The qualitative screening showed bacterial ability to produce hydrogen cyanide, ammonia, siderophore, indole-3-acetic acid (IAA), exopolysaccharide, and solubilize phosphate. The high quantity of siderophore produced by B. cereus T4S was 87.73%. No significant difference was observed in the Bacillus sp. CAL14 (33.83%), S. indicatrix BOVIS40 (32.81%), S. maltophilia JVB5 (32.20%), S. maltophilia PK60 (33.48%), B. subtilis VS52 (33.43%), and P. saponiphilia J4R (33.24%), exhibiting high phosphate-solubilizing potential. S. indicatrix BOVIS40, B. thuringiensis SFL02, B. cereus SFR35, B. cereus BLBS20, and B. albus TSN29 showed high potential for the screened enzymes. Varied IAA production was recorded under optimized conditions. The medium amended with yeast extract yielded high IAA production of 46.43 μg/ml by S. indicatrix BOVIS40. Optimum IAA production of 23.36 and 20.72 μg/ml at 5% sucrose and 3% glucose by S. maltophilia JVB5 and B. cereus T4S were recorded. At pH 7, maximum IAA production of 25.36 μg/ml was obtained by S. indicatrix BOVIS40. All the isolates exhibited high IAA production at temperatures 25, 30, and 37°C. The in vitro seed inoculation enhanced sunflower seedlings compared to the control. Therefore, exploration of copious endophytic bacteria as bioinoculants can best be promising to boost sunflower cultivation.

Morphological and molecular characterization of plant growth promoting salt-tolerant bacteria associated with halophytes in the Southeast seaside of Vietnam

Halophytes are found in high-salt environments naturally, and their roots may be associated with promising microbial candidates for promoting crop growth and salt tolerance. In this study, halotolerant bacteria were isolated from soil and root samples of Rhizophora apiculate (R. apiculate), Avicennia ofcinalis (A. ofcinalis), Thespesia populnea (T. populnea), Acanthus ilicifolius (A. ilicifolius) and Trichophorum cespitosum (T. cespitosum), five native halophytes of southeast seaside of Vietnam. Isolates were tested for maximum salt tolerant and screened for the ability of phosphate solubilization and indole acetic acid (IAA) production. Colony morphology, pigmentation, and Gram staining of each IAA production halotolerant isolate were determined. The bacterial isolates showed the highest salt tolerance and IAA production were identifed by sequencing the 16S rRNA gene. A total of 54 isolates which were able to grow in the presence of up to NaCl 3M were isolated. Twenty-three halotolerant bacterial isolates had the capacity of IAA production, 60.9% from which were Gram positive with a cocci shape, colony in opaque/transparent yellow or opaque/off white, 1 - 2 mm or 2 - 3 mm in diameter with the convex surface. Three isolates VTDD1, VTTD2, and KGOR1 were able to solubilize insoluble phosphorus. The highest IAA production was observed in VTDR1 (93.77 µg/mL) followed by VTMR1 (75.23 µg/mL) and VTDR2 (60.00 µg/mL), while the smallest IAA production was observed in CGOD1 (0.50 µg/mL). The isolates VTDR1 and VTDR2 were identifed as Salinicola tamaricis (99.58% and 99.67% identity respectively), while VTMR1 was found to be Salinicola peritrichatus (98.37% identity).

Isolation and characterization of KDML105 aromatic rice rhizobacteria producing indole-3-acetic acid: impact of organic and conventional paddy rice practices.

Indole-3-acetic acid (IAA) synthesis is a major property of rhizosphere bacteria. The IAA-producing ability of rhizobacteria may be influenced by agricultural management. We therefore evaluated the IAA-producing potential of rhizobacteria isolated during organic rice farming (ORF) and conventional rice farming (CRF) in Thung Kula Rong Hai areas of Thailand. The results indicated that ORF gave a significantly higher percentage of IAA producers (95·8%) than CRF (69·9%). The average IAA values of the ORF isolates were around two times higher than those of the CRF isolates both in the absence (12·8 and 5·8 μg IAA ml-1 , respectively) and presence of L-tryptophan (L-Trp) (35·2 and 17·2 μg IAA ml-1 , respectively). The 16S rRNA gene sequence analysis indicated that the 23 selected isolates belonged to 8 different genera-Sinomonas sp., Micrococcus sp., Microbacterium sp., Fictibacillus sp., Bacillus sp., Burkholderia sp., Leclercia sp. and Enterobacter sp. Interestingly, only three ORF isolates, i.e. ORF15-20 (Micrococcus sp.), ORF15-21 (Sinomonas sp.) and ORF15-23 (Sinomonas sp.), exhibited high IAA production ability without L-Trp (128·5, 160·8 and 174·7 μg IAA ml-1 , respectively). Meanwhile, a slight decrease in IAA production with L-Trp was noticed, suggesting that the L-Trp was not used for the IAA synthesis of these isolates. Biopriming with rhizobacterial isolates significantly enhanced the rate of germination of KDML 105 rice seeds compared to the control.

Indole-3-acetic acid production by rhizobacteria Bacillus spp. to various abiotic stress factors

Indole-3-acetic acid (IAA) phytohormone plays an essential role in forming and initiating main, lateral, and adventitious roots in vegetative propagation. Plants are receiving IAA naturally from a diverse group of soil-plant associated rhizobacteria. However, IAA synthesis by rhizobacteria is influenced by abiotic growth conditions. Three indigenous Bacillus isolates were subject to in vitro assay for the effects of abiotic factors (temperature, salinity and pH) on growth and IAA production. All isolates grew well between 25 - 40°C, and only B. megaterium UPMLH3 was capable of synthesising IAA (21.18 µg/ml) at 40°C. All three bacterial growth under saline stress were slightly dropped over control (0% NaCl), but still producing IAA up to 1% NaCl condition. B. cereus UPMLH24 revealed high resistance to salinity up to 5% NaCl. The optimum growth of all three Bacillus spp. was at pH 7. B. cereus UPMLH1 and UPMLH24 discovered higher IAA production in slightly alkaline conditions (pH 8). Each rhizobacterium shows different physiology trait against each abiotic factor. However, the multiple tolerance ability of PGPR against abiotic factors is an indication that its ability to survive under harsh soil and plant environments while delivering benefits to the plant. Thus, B. cereus UPMLH1, B. megaterium UPMLH3 and B. cereus UPMLH24 might serve as potential biofertiliser, enhancing the growth performance of test plants at various environmental conditions.

Isolation, Morphological and Biochemical Characterization of Rhizobacteria from Arsenic Contaminated Paddy Soils in Bangladesh: An In vitro Study

Soil-plant–microbes relations within the plant rhizosphere are the determinants of plant and soil health, which is important for soil ecological environment for plant-microbe interactions. Plant growth-promoting rhizobacteria (PGPR) are considered to encourage plant growth and development directly or indirectly in soil. PGPR can demonstrate a diversity of characteristics responsible .for influencing plant growth and development. During this study, Twenty four different bacterial isolates were isolated, and detailed morphological, biochemical, and physiological characterizations of those isolates were accomplished. This experiment was performed with the 24 bacterial isolates to see their gram stain test, KOH test, catalase activity, cellulose degradation capability, in dole acetic acid (IAA) production, and phosphate solubilization activities, and also tested for growth within the different arsenic and salt stress conditions and 37°C temperature. Results revealed that among the rhizobacterial isolates, fifteen bacterial isolates were negative and nine was positive in gram reaction, while some were showed high IAA production ability, phosphate solubility capability, and cellulose degradation capacity within the culture media. The isolates were isolated from paddy soils and a few were characterized by a yellow color, flat elevation, and gram-positive, while some were characterized because of the yellowish color with round colony shape, raised elevation, gram-negative, and every one the isolates were positive in catalase production capacity and phosphate solubilization activity which is able to increase the available phosphorus within the soil for plants and also produced indole acetic acid that may use as a hormone to be used in growth enhancement of plants. Hence, these isolates need to be tested further for their effect on arsenic dynamics at the plant rhizosphere, selection of suitable plant species for the bacterial association, bacterial effect on arsenic uptake by plants, and potentials for field applications for sustainable agriculture.

Amelioration of growth attributes of Bambusa nutans subsp. cupulata Stapleton by indole-3-acetic acid extracted from newly isolated Bacillus mesonae MN511751 from rhizosphere of Bambusa tulda Roxburgh

The use of rhizobacteria in sustainable agriculture as biofertilizers and growth promoters has potential to mitigate hazards caused by excessive chemical application. To elucidate the effect of bacterial indole-3-acetic acid (IAA) on growth promotion of bamboo, soil samples were collected from the rhizosphere of Bambusa tulda. Of the 30 bacterial isolates screened by the serial dilution method, 15 isolates were found capable of producing IAA. One of the isolates with high IAA production capacity, phosphate solubilization index, 1-aminocyclopropane-1-carboxylic acid (ACC) deaminase activity was subjected to phylogenetic analysis of 16 S rRNA gene sequence, and was identified as Bacillus mesonae, showing 99% nucleotide similarity to that of B. mesonae FJAT-13985T. IAA extracted from B. mesonae was quantified by high performance liquid chromatography (HPLC) and eluted with the same retention time as standard IAA. A linear relationship between the concentration of standard IAA and the peak area of B. mesonae IAA was observed (r = 0.98), indicating that IAA is present in crude extracts of B. mesonae. The bioactivities of the extracted bacterial IAA and its growth promoting efficiency in seedlings of B. nutans subsp. cupulata were compared with uninoculated control plants, and showed significant promotion of seed germination along with root and shoot development. Therefore, B. mesonae could be a prospective candidate to be employed for the enhancement of growth and development of other bamboo species having high economic and agricultural priority.

Rhizobacteria and Arbuscular Mycorrhizal Fungi of Oil Crops (Physic Nut and Sacha Inchi): A Cultivable-Based Assessment for Abundance, Diversity, and Plant Growth-Promoting Potentials

Nowadays, oil crops are very attractive both for human consumption and biodiesel production; however, little is known about their commensal rhizosphere microbes. In this study, rhizosphere samples were collected from physic nut and sacha inchi plants grown in several areas of Thailand. Rhizobacteria, cultivable in nitrogen-free media, and arbuscular mycorrhizal (AM) fungi were isolated and examined for abundance, diversity, and plant growth-promoting activities (indole-3-acetic acid (IAA) and siderophore production, nitrogen fixation, and phosphate solubilization). Results showed that only the AM spore amount was affected by plant species and soil features. Considering rhizobacterial diversity, two classes—Alphaproteobacteria (Ensifer sp. and Agrobacterium sp.) and Gammaproteobacteria (Raoultella sp. and Pseudomonas spp.)—were identified in physic nut rhizosphere, and three classes; Actinobacteria (Microbacterium sp.), Betaproteobacteria (Burkholderia sp.) and Gammaproteobacteria (Pantoea sp.) were identified in the sacha inchi rhizosphere. Considering AM fungal diversity, four genera were identified (Acaulospora, Claroideoglomus, Glomus, and Funneliformis) in sacha inchi rhizospheres and two genera (Acaulospora and Glomus) in physic nut rhizospheres. The rhizobacteria with the highest IAA production and AM spores with the highest root-colonizing ability were identified, and the best ones (Ensifer sp. CM1-RB003 and Acaulospora sp. CM2-AMA3 for physic nut, and Pantoea sp. CR1-RB056 and Funneliformis sp. CR2-AMF1 for sacha inchi) were evaluated in pot experiments alone and in a consortium in comparison with a non-inoculated control. The microbial treatments increased the length and the diameter of stems and the chlorophyll content in both the crops. CM1-RB003 and CR1-RB056 also increased the number of leaves in sacha inchi. Interestingly, in physic nut, the consortium increased AM fungal root colonization and the numbers of offspring AM spores in comparison with those observed in sacha inchi. Our findings proved that AM fungal abundance and diversity likely rely on plant species and soil features. In addition, pot experiments showed that rhizosphere microorganisms were the key players in the development and growth of physic nut and sacha inchi.

Deciphering operation of tryptophan-independent pathway in high indole-3-acetic acid (IAA) producing Micrococcus aloeverae DCB-20.

Genus Micrococcus is considered a high IAA producer. However, interestingly, there is no report on the tryptophan- independent pathway operation in this genus. Consequently, the present study was undertaken to evaluate high IAA production by Micrococcus aloeverae DCB-20 and generate reasonable evidence for the occurrence of the tryptophan-independent pathway. Strain DCB-20 produced a high quantity of 880.51 µM or 154.3µg/mL IAA in LB broth supplemented with L-tryptophan. The tryptophan-independent pathway operation was supported by IAA production in Tris-minimal broth (TM broth) medium supplemented with acid hydrolyzed casein hydrolysate (casein acid hydolysate), which lacks tryptophan. The HPLC analysis showed the absence of tryptophan either from exogenous or endogenous sources in TM broth in the presence of casein acid hydrolysate inoculated with M. aloeverae DCB-20. The absence of tryptophan was further confirmed by the appearance of non-pigmented colonies of Chromobacterium violaceum strain TRFM-24 on Tris-minimal agar (TM agar) containing acid-hydrolyzed casein. This is probably the first report on IAA biosynthesis by M. aloeverae DCB-20 employing tryptophan-independent pathway. This simple technique can also be adapted to detect operation of the tryptophan-independent pathway in other bacteria.

Extraction of Indole-3-acetic Acid from Plant Growth Promoting Rhizobacteria of Bamboo Rhizosphere and Its Effect on Biosynthesis of Chlorophyll in Bamboo Seedlings

Background: Indole-3-acetic acid (IAA), a principal phytohormone, controls several crucial physiological processes of plants. It ameliorates plant growth by stimulating cell elongation, root initiation, seed germination and seedling growth. Alteration of IAA level by plant growth promoting rhizobacteria leads to varied impacts on plant growth and development. Methods: Soil samples were collected from bamboo (Bambusa tulda, B. nutans subsp. cupulata, B. balcooa and Dendrocalamus strictus) rhizosphere. Altogether five bacterial isolates were screened by serial dilution method and subjected to biochemical analysis. The isolate BUX1 with high IAA production capacity was optimized for IAA production. IAA was partially purified and quantified from the bacterial extract by thin layer chromatography (TLC). The influence of extracted bacterial IAA on chlorophyll biosynthesis in bamboo seedlings of B. tulda was compared with uninoculated control plants. Results: Biochemical analysis revealed that all the isolates belonged to genus Bacillus which were found capable of producing IAA. During optimization, BUX1 isolate produced 99.13 µg ml-1 of IAA at 37°C, pH 7, 3 mg l-1 concentration of L-tryptophan and 150 rpm agitation rate after 192 hour of incubation. The Rf value of the bacterial IAA during TLC was identical to that of standard IAA (0.425) indicating that IAA was present in crude extract of Bacillus (BUX1). The influence of bacterial IAA on chlorophyll biosynthesis in bamboo seedlings was significant in comparison to uninoculated plants. Therefore, this isolate could be a prospective candidate to be employed as biofertilizer.

Bioremediation potential of new cadmium, chromium, and nickel-resistant bacteria isolated from tropical agricultural soil

Soil management using fertilizers can modify soil chemical, biochemical and biological properties, including the concentration of trace-elements as cadmium (Cd), chromium (Cd) and nickel (Ni). Bacterial isolates from Cd, Cr, and Ni-contaminated soil were evaluated for some characteristics for their use in bioremediation. Isolates (592) were obtained from soil samples (19) of three areas used in three maize cultivation systems: no-tillage and conventional tillage with the application of mineral fertilizers; minimum tillage with the application of sewage sludge. Four isolates were resistant to Cr3+ (3.06 mmol dm−3) and Cd2+ (2.92 mmol dm−3). One isolate was resistant to the three metals at 0.95 mmol dm−3. All isolates developed in a medium of Cd2+, Cr3+ and Ni2+ at 0.5 mmol dm−3, and removed Cd2+ (17–33%) and Cr6+ (60–70%). They were identified by sequencing of the gene 16S rRNA, as bacteria of the genera Paenibacillus, Burkholderia, Ensifer, and two Cupriavidus. One of the Cupriavidus isolate was able to remove 60% of Cr6+ from the culture medium and showed high indole acetic acid production capacity. We evaluated it in a microbe-plant system that could potentially be deployed in bioremediation by removing toxic metals from contaminated soil.

IAA production and phosphate solubilization performed by native rhizobacteria in western Paraná

In search for a more sustainable agriculture, the use of microorganisms as a technology is increasingly being used by agriculture throughout the world. This is due to the fact that it minimizes the use of agricultural supplies reducing environmental costs and impacts, based on the beneficial and natural relationships between edaphic organisms and cultivated plants. The rhizobacteria habitat in the soil establishes biochemical relationships with the plants acting as plant growth promoters (PGPR). Many of these bacteria are producers of phytohormones and enzymatic compounds with the capacity to provide important nutrients for plants. In this context, the present work aimed to quantify the potential of indole-3-acetic acid (IAA) production and the phosphate solubilization of rhizobacteria from Western Paraná. Isolates grown in DYGS medium plus tryptophan were quantified by colorimetry for the production of IAA. Iron phosphate solubilization was carried out by inoculation in modified Pikovskaya medium (PKV) and quantified by colorimetry. The results were evaluated by the Scott-Knott test at 5% using the SASM-Agri program. The highest IAA production was observed with the addition of tryptophan to Erwinia (219); Enterobacter (302) and Salmonella (57). Isolates Falsibacillus (438) and 505 showed higher efficiency in the iron phosphate solubilization. Isolates Enterobacter (130), 438 and Enterobacter (151) were highlighted in both tests, being characterized as a great potential for use in biotechnological products.

PHYSIOLOGICAL CHARACTERIZATION OF DIAZOTROPHIC BACTERIA ISOLATED FROM Brachiaria brizantha RHIZOSPHERE

ABSTRACT The objective of this study was to evaluate the ability of diazotrophic bacteria isolated from Brachiaria brizantha rhizosphere to grow at different pH values and salt concentrations, to produce indoleacetic acid (IAA), and to solubilize phosphate. Both acidity and salinity tolerance tests were performed on modified solid FAM medium. Acidity resistance tests were performed at pH values of 4.0, 5.0, 6.0, and 7.0, while, salt resistance was evaluated at concentrations of 0.01, 5.02, 9.99, 14.96, 19.98, 24.95, 29.98, 34.94, 39.97, 44.94, and 49.96 gL-1 NaCl. For the IAA production assay, bacterial strains were cultivated in liquid DYGS medium both in the absence and presence of tryptophan (Trp). In addition, phosphorus solubilization assay was performed in GL liquid medium. All strains grew at every pH value tested, and a high diversity was observed after salt resistance, IAA production, and phosphate solubilization testing. Strains UNIFENAS 100-51, UNIFENAS 100-52, UNIFENAS 100-60, UNIFENAS 100-63, and UNIFENAS 100-65 were those with the best growth at the highest salt concentrations. Furthermore, in the presence of Trp, strains UNIFENAS 100-63 and UNIFENAS 100-69 were the ones with the highest IAA production. Strain UNIFENAS 100-52 showed the best response to the in vitro phosphate solubilization assay. Based on these results, it can be seen that studies related to the physiological and metabolic characteristics of diazotrophic bacterial strains are important to ensure greater success in the field.