ACCENT JOURNAL OF ECONOMICS ECOLOGY & ENGINEERING Available Online: www.ajeee.co.in Vol. 01, Issue 05, September 2016, ISSN -2456-1037 (INTERNATIONAL JOURNAL)
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AN APPLICATION OF PHOSPHATE SOLUBILIZATION BY RHIZOBIUM STRAINS: A STUDY
Dr. Bharat Jinturkar,
Head Department of Botany and Principal, Late K.G. Kataria College Daund, Pune, Maharashtra, India
Abstract:- Forty-six Rhizobium isolates from legume root and stem nodules were examined for their phosphate-solubilizing ability on Pikovskaya’s agar medium. Rhizobium isolates from root nodules of Cassia absus, Vigna trilobata and three lines from Sesbania sesban confirmed quarter of tricalcium phosphate (TCP) solubilization. The isolate from C. Absus confirmed maximum solubilization (620μg/ml) after 12d of incubation, whilst the Rhizobium sp. Pressure 26 (from S. Sesban) confirmed the least amount (150μg/ml) of phosphate solubilization. Some of the carbon resources tested for his or her potential to solubilize TCP, maximum solubilization (620μg/ml) was found in glucose by way of Rhizobium isolate from C. Absus. Phosphate solubilization multiplied with boom in glucose attention steeply up to 2% and slowly above this awareness in four isolates. A number of the nitrogen resources tested, maximum solubilization (620μg/ml) became observed in ammonium sulphate by way of Rhizobium isolate from C. Absus.
Keywords:- Phosphate solubilization, Rhizobium, Tricalcium phosphate.
1. INTRODUCTION
Availability of phosphate in soil is substantially greater through microbial manufacturing of metabolites main to lowering of pH and release of phosphate from natural and inorganic complexes.
Phosphorus (P) deficiency in soil can significantly restriction plant boom productiveness, mainly in legumes, wherein each the vegetation and their symbiotic micro organism are affected, and this could have a deleterious impact on nodule formation, improvement and characteristic.
Except, symbiotic nitrogen fixation, a few strains or species of Rhizobium are concerned in phosphate solubilization additionally. However studies on phosphate solubilizing potential of Rhizobium strains are very restrained. The main advantage of using rhizobia as a phosphate-solubilizing microorganism will be their beneficial nutritional impact resulting each from phosphate mobilization and nitrogen fixation. In the gift have a look at, 46 rhizobia, remoted from root and stem nodules of 20 different legume hosts, have been examined for their capacity to solubilize tricalcium phosphate (TCP).
Of the forty six isolates, 26 are from Sesbania sesban and the last 20 isolates consist of one each from Abrus precatoreus, Arachis hypogaea, Cajanus cajan, Cassia absus, Clitorea ternatea, Dolichos lablab, Indigofera hirsuta, I.
Linnaei, I. Trita, Leucaena leucocephala, Macrotyloma unifl orum, Pisum sativum,
Rhynchosia minima, Sesbania grandifl ora, S. Procumbens (root), S. Procumbens (stem), Trigonella foenum-graecum, Vigna mungo, V. Radiata and V. Trilobata.
The selective medium yeast extract mannitol agar (YEMA) with congo red became used for isolation of rhizobia and a pure culture of each isolate became prepared after sub culturing on the equal medium. Natural cultures were authenticated as rhizobia thru biochemical exams and nodulating potential became tested on homologous hosts with the aid of plant contamination checks. A representative isolate from S.
Sesban changed into identified as Rhizobium radio bacter MTCC 8917 (=Agrobacterium radiobacter).
On account that Agrobacterium and Rhizobium are still treated as separate genera in Bergey’s guide of Systematic Bacteriology, we used the term Rhizobium sp. With stress numbers as 1–
26. The alternative 20 root nodule isolates were special with the aid of the not unusual time period rhizobia with host call in parenthesis. The phosphatesolubilizing potential of the isolates changed into tested on Pikovskaya’s agar medium containing TCP as insoluble phosphate source. The solubilization efficiency (SE) on agar medium becomes expressed in phrases of SE (%).
The isolates, which showed quarter of solubilization on agar medium changed into further examined in fl asks
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2 containing a hundred ml of Pikovskaya’s broth having initial pH 7.Zero. The broth become inoculated with 1 ml of the inoculums and incubated at 28 ± 2°C on rotary shaker (200 rpm) for 12 d. 10 ml aliquots of each tradition have been aseptically taken from every fl ask after three d intervals up to 12 d, and the supernatant was separated from bacterial cells via centrifugation (3000 g). The final pH of the supernatant was measured and the liberated P2 O5 became estimated.
A few of the 46 Rhizobium isolates, simplest five isolates, one each from C.
Absus and V. Trilobata and 3 lines from S. Sesban (Rhizobium sp. Strain 17, 19 and 26) produced clean zone around the colonies after three d of incubation, and it step by step accelerated as much as nine d, at the same time as the colony diameter is nearly similar at some stage in the incubation duration. Solubilization efficiency (SE) of Rhizobium isolates on stable media ranged between 33% and 150%).
Most performance turned into observed in Rhizobium isolate from C.
Absus (150%) observed via S. Sesban Rhizobium sp. Pressure 17 (125%), Rhizobium sp. Stress 19 (120%), Rhizobium sp. Strain 26 (100%) and Rhizobium isolate from V. Trilobata (60%).
In liquid medium maximum solubilization become recorded with Rhizobium isolate from C. Absus (620 μg/ml) accompanied with the aid of Rhizobium sp. Strain 19 (391 μg/ml), and least in Rhizobium sp.
Stress 26 (156 μg/ml) from S. Sesban.
It became also determined that there is no correlation among phosphate solubilization performance on strong and liquid medium as additionally noticed in advance. A fall in pH followed phosphate solubilization, because of manufacturing of organic acids become determined up to 9 d, however there has been sudden boom in pH after 9 d. This may be due to utilization of natural acids by the strains as turned into stated in Pseudomonas.
The information had been statistically analyzed the use of correlation coefficient and it turned into located that there is tremendous correlation between area of solubilization on Pikovskaya’s agar medium and liberated P2 O5 in broth, and terrible correlation among liberated P2 O5 and final pH of the medium in broth. Effect of different carbon resources (1%) on
phosphate solubilizing interest become studied via replacing the glucose with 14 carbon compounds sterilized one at a time and delivered aseptically to the fl asks containing 100 ml of the Pikovskaya’s medium earlier than inoculation.
The medium was incubated on a rotary shaker (2 hundred rpm) at room temperature (28 ± 2°C) for 72 h. Most of the 14 carbon resources examined, glucose supported maximum TCP solubilization in Rhizobium isolate from C. Absus (620 μg/ml), accompanied by way of Rhizobium sp. Pressure 19 (390 μg/ml), Rhizobium sp. Strain 17 (381 μg/ml) and Rhizobium sp. Strain 26 (155 μg/ml). While the isolate from V. Trilobata showed most solubilization (466 μg/ml) in fructose.
That glucose becomes the pleasant carbon source for phosphate solubilization changed into additionally said in advance in Bradyrhizobium species isolated from Cicer arietinum. The most decrease in pH was recorded in glucose-containing medium. In different carbon sources little lower in pH and no correlation between acidic pH and amount of P2 O5 liberated had been determined. Those consequences indicated that mannitol was no longer a very good carbon supply for phosphate solubilization research in rhizobia, and therefore all assessments had been achieved with Pikovskaya’s medium containing glucose as carbon supply.
The relative performance of Rhizobium isolates on different carbon sources could be because of the organic acids secreted by the isolates instead of the overall acidity. The statistics on impact of carbon resources on TCP solubilization via Rhizobium lines had been statistically analyzed the use of evaluation of variance (-way classification method) and it turned into located that versions due to both carbon sources and Rhizobium strains have been located to be signifi cant.
Because the glucose at 1%
awareness (as in Pikovskaya’s medium) supported most solubilization of TCP, effect of various concentrations of glucose (1, 1.5, 2, 2. 5, 3 %) turned into studied.
Phosphate solubilization expanded with increase in glucose attention in Rhizobium isolates from C. Absus, V.
Trilobata and Rhizobium sp. Strain 17
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3 and 19 from S. Sesban. It became also discovered that boom in glucose awareness from 2% to 3% did not show plenty change in TCP solubilization.
Inside the case of Rhizobium sp.
Pressure 26, maximum solubilization turned into discovered at 1.5% glucose concentration and above this concentration TCP solubilization decreased. This may be because of the auto consumption of soluble phosphate by the growing bacterial populace as pronounced in Azospirillum brasilense.
The version within the effect of different concentrations of glucose is discovered to be statistically sizeable inside the absence of NH4 + additionally influences phosphate solubilization effectively.
Of all of the isolates, Rhizobium isolate from C. Abuses showed maximum solubilization (620μg/ml) in ammonium sulphate containing medium, at the same time as least solubilization (12μg/ml) turned into discovered in urea with the aid of Rhizobium sp. Stress 26. Further it was discovered that inorganic nitrogen assets supported better solubilization of TCP than natural nitrogen resources. This will be due to the production of inorganic acids with the aid of proton alternate mechanism inside the presence of NH4 + motive expanded phosphate solubilization.
Statistical evaluation confirmed that the impact of different nitrogen sources on TCP solubilization changed into also vast. From this take a look at it is clear that phosphate solubilization isn't always a substantial individual among rhizobia, and not all the Rhizobium traces, even from a single host (S.
Sesban), showcase phosphate solubilization. For this reason, it's far important to look at the phosphate- solubilizing pastime of a big range of Rhizobium traces from an unmarried host.
Furthermore, the Rhizobium strains displaying phosphate solubilization may be used for information the mechanism of phosphate acquisition in host flowers from which they were remote.
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