Final SPH -JHS Coverpage 17-1 Jan 2022 single 131 J. Hortl. Sci. Vol. 17(1) : 131-136, 2022 This is an open access article d istributed under the terms of Creative Commons Attribution-NonCommer cial-ShareAl ike 4.0 International License, which permits unrestricted non-commercial use, d istribution, and reproduction in any med ium, provide d the original author and source are credited. Original Research Paper INTRODUCTION Soil utilitarian bacteria act as a significant contributing factor for improving diminished soil fertility. Soil bacteria increase the crop production through their inherent characteristics of soil structure improvisation, production of hormones and enzymes, controlling pathogen and heavy metal. Nevertheless, the decline of orga nic fer tilizer s usa ge a nd soil fer tility exploita tion thr ough a gr ochemica l a ffects the horticultural crop yield rate (Malhotra et al., 2017). The incorporation of soil utilitarian bacteria as biofertilizers is needed to overcome the aforementioned issues. Statistical infusion in the soil nutrient and bacterial data analysis would bring out a realistic field study. From the extensive works of literature, it is found that various initiatives have been taken to improvise horticultural crops productivity through soil nutrient and bacterial analysis. Besides, beneficiary microbes such as A. chroococcum, A. vinelandii, A. beijerinckii, A. paspali, A. armeniacus, A. nigricans, A. salinestri , Azospirillum, Cyanobacteria, Azolla, Gluconacetobacter diazotrophicus, Bacillus aerius, Bacillus amyloliquefaciens, Bacillus mucilaginous, Bacillus subtilis, Enterobacter contributes for nitrogen fixation in the soil (Dhayalan and Karuppasamy 2021). Pseudomonas chlororaphiswere, P. putida, P. entomophila, P.koreensis, P.luteola, P. simiae, P.stutzeri, Bacillus sp., Achromobacter, Acinetobacter, Aeromonas hydrophila, Arthroderma cuniculi, Aspergillus niger, Bacillus aerius, B. altitudinis, B. thuringenesis, Enterococcus casseliflavus, E. gallinarum, Lecanicillium psalliotae, Paenibacillus taichungensis, Serratia nematodiphila, Sphingomonas paucimobilis, Azotobacter etc. are some of the soil phosphorus solubilizing microbes which are used in different horticultural crops for enhanced productivity (Mussa et al., 2018). Pseudomonas azotoformans, Burkholderia, Bacillus mucilaginosus, B. edaphicus, B. circulans, Pseudomonas, Acidithiobacillus ferrooxidans, Paenibacillus sp., and Enterobacter hormoecheiin are some of the beneficiary bacteria that induces the solubilization of potassium (Prajapati and Modi 2016). Correlation among the soil nutrients and their associated bacterial genera is a substantial research gap. Resultant of correlation leads gateway for precise decision making relevant to bio fertilization for horticultural crops. Macronutrients and their associated bacterial genera in the soils of Anaimalai block in Tamil Nadu for sustainable vegetable crops cultivation Dhayalan V. and Sudalaimuthu K.* Department of Civil Engineering, SRM Institute of Science and Technology, Kattankulathur, Tamilnadu, India *Corresponding author Email: karuppas@srmist.edu.in ABSTRACT Meticulous analysis on intertwined interaction among the soil nutrients and microbial communities brings fruitful outcomes on horticulture. This study focuses on identifying well compatible bacterial genera in enhancing soil primary macro-nutrients for sustainable vegetable crops cultivation. The biochemical tests were executed for identification of bacterial genera. Eventually, mathematical models among available NPK nutrients (Nitrogen, Phosphorus and Potassium) and identified bacterial genera were derived to determine most suited bacterial genus for nutrients inhibition. This Study reveals the present nutrient’s status of soils at Anaimalai block covering 12,424 ha of Coimbatore district in Tamil Nadu. Seven utilitarian bacterial genera were identified which inhibit the plant nutrients. Among them, Azotobacter, Arthrobacter and Achromobacter actively inhibit available NPK in the soil. Present study of correlating soil nutrients with bacterial components enriches successful conservation of biosphere through adopting these innovative technologies in horticulture. Keywords: Correlation, horticultural crops, macronutrients, soil beneficiary microbes, soil fertility, sustainable agriculture and vegetable production. 132 Dhayalan and Sudalaimuthu J. Hortl. Sci. Vol. 17(1) : 131-136, 2022 MATERIALS AND METHODS Sample site description Extensive research work was performed recently in Anaimalai at Pollachi, which lies between 10.662°N 77.00650°E, located 40km to the south of Coimbatore, India. The study area consists of vegetable crops such as tomato, bhendi, brinjal, chilies, caspium, paprika, pumpkin, snake gourd, bitter gourd, cluster bean, potato, cabbage, cauliflower, onion etc., Twenty-seven soil samples were collected from the rhizospheric region of horticultural crops for macronutrients analysis. Macronutrient analysis The basic physical parameters such as pH, Electrical conductivity, and salinity were measured using pH meter (Elico LI 617), conductivity meter (Elico CM 183), and salinity meter respectively. Available nitrogen in the soil sample was analyzed using the Kjeldahl nitrogen analysis method. Total phosphorus was analyzed using the Bray-1 method for acidic soil samples and Olsen method for alkaline soil samples. Available potassium in the soil was estimated using Flame Photometer (Jenway PFP7). Bacterial analysis Enumeration of microorganism Enumeration of bacteria were carried out through serial dilution method to reduce the microbial load and plated by pour plate method. Bacteria being isolated fr om the pla tes a r e incuba ted a t 37°C in a bacteriological incubator. Gram staining Gram staining was done for the isolated bacterial cultures in a slide using Gram’s iodine, decolorizing agent and safronin strain. The slides were observed under the trinocular microscope, the purple colors indicated gram positive bacteria reveals the presence of bacterial genera, Arthrobacter and most of slides obser ved pink color indicating the pr esence of Azotobacter, Enterobacter, Citrobacter, Achromobacter, E. coli and Cornybacterium which are characterized by gram negative. Further analyses were carried out to confirm the presence of initially traced bacterial genera. The cover slip was taken where its edge was coated with vaseline and the test samples were transferred to the cover slip which was placed over the cavity slide. The slide was viewed under 100X ma gnifica tion and the orga nisms’ characteristics being motile or non-motile were noted down. Biochemical test for bacterial genera For the confirmation of bacterial genera certain biochemical tests were carried. Indole test were carried out for the isolated bacteria culture in which a red colour ring or pink colour ring at the top reviling positive reaction, where yellow colour ring at the top indicate negative reaction. Indole test reveals that most of the identified bacterial genera except Arthrobacter, Citrobacter and Enterobacter exhibit the positive reaction. Followed by the indole test, methyl red test were performed, Formations of red colour ring at the top indicate the positive reaction which reveals the ex is t enc e of A z o t o b a c t e r, C i t ro b a c t e r, Achromobacter, E.coli and Cornybacterium and formation of yellow colour ring at top shows the negative reaction which indicate the presence of A r t h ro b a c t e r a nd E n t e ro b a c t e r . F u r t her confir ma tion of bacter ia l gener a was made by Simmons citrate utilization test, so as to determine the ability of the microorganism to use citrate as its sole source of carbon. From the Simmons citrate utilization test it was revealed that the survival of A zo to ba ct er , Ci trob a ct er , E nt ero ba ct e r an d Achromobacter were reverbera ted by means of gr een to blue colour cha nges in the ba cter ia l medium specifying positive reaction. The existence of b a c t e r ia l gener a s u c h a s A r t h ro b a c t e r, Cornybacterium and E.coli were revealed through green to yellow color changes of bacterial medium indicating the negative reaction (Ishiguro et al., 1979). Voges Pr oska uer tests were included to analyze the presence of identified bacterial genera at most precise manner. Voges Proskauer tests are used to demonstrate an organism’s ability to convert pyruvate to acetoin. Formation of red colour ring exhibits the positive reaction which covey the existence of Enterobacter genera and yellow colour ring express the negative reaction exposing the s u r viva l of A z o t o b a c t e r, C i t ro b a c t e r, Achromobac ter, E. coli, Cornyba cterium, a nd Arthrobacter. Bacterial genus interpretation Based on the outcomes of va rious biochemical tests, identifica tion of bacter ia were made with 133 Macronutrients and their associated bacterial genera t h e he lp of B er g ey ’s ma nu a l of s ys t ema t ic ba cteriology. Out of 11,669 different bacterial c o l o n i e s 3 1 % o f A r t h ro b a c t e r , 2 4 % o f Citrobacter, 17% of Escherichia coli, 13% of Azotob acter, 7 % of Enterob acter a nd 5% of A c h ro m o b a c t e r , wa s f ou nd a s c on t r ib u t i ng ba cter ium for pr omoting soil nutr ients. Tota l colonies of bacter ia l genus wer e identified by multiplying the average number of colonies with 105 as dilution factor (Ta ble 1). Table 1. Biochemical characteristic for the identification of bacterial species S.No. S.F. No Latitude Longitude Total BC GS IT MRT SCUT VPT IB CC 1 590 10.588153 76.885931 380 GNR (+) (+) (+) (-) Azotobacter 157 2 592 10.587975 76.885421 200 GNR (-) (-) (+) (+) Enterobacter 125 3 596 10.587718 76.885257 511 GNR (-) (+) (+) (-) Citrobacter 425 4 600 10.588111 76.883713 561 GNR (+) (+) (+) (-) Achromobacter 362 5 601 10.587909 76.884139 356 GNR (+) (+) (-) (-) E.coli 208 6 602 10.587721 76.884218 312 GPR (-) (+) (-) (-) Cornybacterium 215 7 606 10.587537 76.884144 326 GNR (-) (-) (+) (+) Enterobacter 225 8 608 10.587163 76.884039 322 GPR (-) (-) (-) (-) Arthrobacter 253 9 630 10.586454 76.882942 320 GNR (+) (+) (+) (-) Azotobacter 257 10 637 10.587134 76.883235 320 GNR (+) (+) (-) (-) E.coli 218 11 638 10.587643 76.883528 320 GNR (+) (+) (-) (-) E.coli 218 12 641 10.587063 76.88287 312 GNR (+) (+) (-) (-) E.coli 218 13 642 10.587491 76.882627 245 GPR (-) (-) (-) (-) Arthrobacter 253 14 648 10.587277 76.882267 478 GNR (-) (+) (+) (-) Citrobacter 425 15 649 10.587783 76.88216 520 GNR (-) (+) (+) (-) Citrobacter 425 16 657 10.587926 76.882583 956 GNR (+) (+) (+) (-) Azotobacter 557 17 656 10.588503 76.882551 288 GNR (-) (-) (+) (+) Enterobacter 225 18 660 10.588254 76.883272 540 GPR (-) (-) (-) (-) Arthrobacter 253 19 661 10.588261 76.883422 300 GNR (+) (+) (-) (-) E.coli 218 20 702 10.587996 76.87914 96 GNR (+) (+) (-) (-) E.coli 58 21 703 10.588698 76.87921 96 GNR (+) (+) (-) (-) E.coli 68 22 721 10.589401 76.879324 96 GNR (-) (-) (+) (+) Enterobacter 55 23 747 10.588187 76.877757 87 GNR (-) (-) (+) (+) Enterobacter 72 24 750 10.587696 76.877974 956 GPR (-) (-) (-) (-) Arthrobacter 553 25 751 10.587271 76.878479 956 GPR (-) (-) (-) (-) Arthrobacter 651 26 756 10.587306 76.878045 956 GPR (-) (-) (-) (-) Arthrobacter 253 27 758 10.587714 76.877569 859 GNR (-) (+) (+) (-) Citrobacter 425 BC, Bacterial colonies; GS, Gram Straining; GNR, Gram negative- Rod; GPR, Gram positive- Rod; IT, Indole test; MRT, Methyl red test; SCUT, Simmons Citrate Utilization test; VPT, Voges Proskauer Test; IB, Identified Bacteria; CC, Colonies Count; (+), Positive; (-), Negative. J. Hortl. Sci. Vol. 17(1) : 131-136, 2022 134 RESULTS AND DISCUSSION Statistical Analysis The resulted characteristics of soil were compared with TNAU standards. Pearson coefficients (Table 2.) were incorporated which represents the relationship among the nutrients measured and the associated microbes. Statistics which includes bacterial symmetry, bacterial counts, mean value, minimum bacterial count were interpreted by descriptive statistical analysis (Table 3). Physical parameters and primary macronutrients The study area is observed with clay loam, sandy clay loam and sandy loam types of soil. Loam soil is the mixture of sand, silt and clay having the pH value 4.5 to 6.5. Most of samples in the study area are reddish brown in color indicating fine soil texture that greatly helps in the horticulture. Soil pH value at Anaimalai ranges from 6.41 to 8.72. The overall pH result make a strong report that the pH values which falls above 6.5 in some field area is completely due to the other predominant factors such as water and agrochemicals. The bacterium plays a key role in maintaining the soil pH range (Hoorman, 2016). One of the identified genera Citrobacter count holds positive correlation with pH (Table 2). Citrobacter can maintain the soil pH ranges from 3 to 11 (Oliveira et al., 2016). Electrical conductivity finds its value ranges from 0.08 to 0.9 and the highest EC recorded is 0.9 dS/m. Excess salinity causes a huge hindrance for horticulture (Habib et al., 2016). Beneficial bacteria lower the concentration of ethylene that directs to deduce the salinity stresses in horticulture farmlands. Utilitarian bacterial genuses identified were Achromobacter and Azotobacter which are positively correlated (Table 2) with Electrical conductivity. Field available nitrogen ranges from 128 to 265 kg/ha. Inspite of two bacterial Table 2. Pearson correlation among soil available NPK and bacterial species Pearson Correlations Nitrogen Phosphorus Potassium AC AR AZ CI CO E.Coli EN Nitrogen 1 0.291 -0.405* -0.097 -0.366 -0.016 -0.188 0.077 0.150 0.276 Phosphorus 0.291 1 0.322 -0.273 0.175 0.357 -0.239 -0.081 -0.059 0.003 Potassium -0.405* 0.322 1 0.121 -0.170 -0.031 -0.058 0.186 -0.145 0.136 AC -0.987 -0.273 0.121 1 -0.093 -0.061 -0.082 -0.038 -0.105 -0.081 AR -0.366 0.175 -0.175 -0.093 1 -0.147 -0.198 -0.093 -0.255 -0.197 AZ -0.016 0.357 -0.031 -0.061 -0.147 1 -0.129 -0.061 -0.166 -0.128 CI -0.118 -0.239 -0.058 -0.082 -0.198 -0.129 1 -0.082 -0.224 -0.172 CO 0.077 -0.081 0.186 -0.038 -0.093 -0.061 -0.082 1 -0.105 -0.081 E.Coli 0.150 -0.059 -0.145 -0.105 -0.255 -0.166 -0.224 -0.105 1 -0.222 EN 0.276 0.003 0.136 -0.081 -0.197 -0.128 -0.172 -0.081 -0.222 1 *. Correlation is significant at the 0.05 level (2-tailed). AC, Achromobacter; AR, Arthrobacter; AZ, Azotobacter; CI, Citrobacter; CO, Cornybacterium; E.Coli, Escherichia Coli; EN, Enterobacter; N total Mean SD Sum Skewness Kurtosis CV Min Median Max Achromobacter 27 13.40 69.66 362 5.19 27 5.19 0 0 362 Arthrobacter 27 82.07 175.8 2216 2.26 4.64 2.14 0 0 651 Azotobacter 27 35.96 118.6 971 3.80 15.28 3.29 0 0 557 Citrobacter 27 62.96 153.8 1700 2.09 2.59 2.44 0 0 425 Cornybacterium 27 7.96 41.37 215 5.19 27 5.19 0 0 215 E.Coli 27 44.66 84.91 1206 1.58 0.72 1.09 0 0 218 Enterobacter 27 26 64.10 702 2.59 5.94 2.46 0 0 225 SD, Standard Deviation; CV, Coefficient of Variation; Min, Minimum; Max, Maximum; Table 3. Descriptive analysis of bacterial species. Dhayalan and Sudalaimuthu J. Hortl. Sci. Vol. 17(1) : 131-136, 2022 135 genus presences in the soil, soil available nitrogen indicates low status. This is due to the presence of E. coli which is not a nitrogen fixation bacterium and also due to other environmental factors. Nevertheless, the outcomes of the genus action in the soil still contribute 19% of available nitrogen to the plants. Available phosphorus ranges from 15 to 37 kg/ha in the field and fulfills the recommended level which is considered as major growth contributing factor in horticultural crops productivity. Two major bacterial genera Arthrobacter and azotobacter shows high positive corr elation with phosphor us (Ta ble 2).Var ious literatures show the importance of azotobacter and Arthrobacter in solubilizing the soil phosphorus (Banerjee et al., 2010). Potassium founds to be extracted at higher concentration by horticultural crops (Pimentel et al., 2015) which are in the range between 132 to 374 kg/ha. Soil bacteria ensure prolonged crop growth through its production of inorganic acids, acidolysis, polysaccharides and chelation (Archana et al., 2013). Contributing identified bacterial genus Achromobacter and Enterobacter shows positive correlation with available potassium level (Table 2). Identified Bacterial genera Ma ximum of 362 colonies wer e found to be Achromobacter genus, which gives positive correlation with soil Electrical conductivity and Potassium. Achromobacter genus is capable of solubilizing 5.4 µg/ml of potassium in the soil at maximum extend (Gupta et al., 2016). Nevertheless, the genus finds negative correlations with nitrogen and phosphorus. Achromobacter induce ethylene hor mones for contribution of soil available nitrogen (Bangash et al., 2021) and helps to expose to transient water stress in horticultural crops more specifically tomato and pepper. Arthrobacter genus ranges maximum of 651 in terms of colonies count at the selected study boundary. Distinctive nature of Arthrobacter in synthesizing plant hormones alleviates the phosphorus deficiency and stress developed by the salinity in the soil (Etesami and Glick, 2020). The genus finds positive correlation with soil phosphorus whereas shows negative correlation with other soil nutrients. Arthrobacter genus induces active mechanism against salt stress in the Pisum sativum (pea) crops. Maximum of 557 Azotobacter colonies count were identified which implies positive correlations with phosphorus, whereas negative correlation with other soil nutrients. Though azotobacter magnifies the nitrogen fixation in the soil through active production of phytopathogenic inhibitors, mathematical model implies negative correlation because of the presence of toxic inorganic fertilizers. Azotobacter due to its biological activity helps in increasing the phosphorus solubilization. Compared to Arthrobacter, azotobacter implies the indole a cidic a cids which a r e r esponsible for phosphorus solubiliza tion (Aung et al., 2020). Citrobacter (425 colonies count) maintain the soil pH in recommended range because of its incredible biological activity (Oliveira et al., 2016). Maximum of 218 colonies were found to be Escherichia coli genera, which give positive correlation with soil nitrogen and negative correlations with other soil nutrients. E.coli is not a nitrogen fixing bacterium but possibly helps in nitrogen cycle by producing urea when E. coli utilizes ammonium. Potentiality of E. coli in producing iron-chelating compounds helps to promote the growth of potato crops. Enterobacter due to ACC deaminase activity induces nitrogen and potassium in the soil (Guo et al., 2020). Production of indole-3-acetic acids by the Enterobacter helps in phosphorus solubilization thus in turn multiple the yields of tomato, cucumber and pepper crops. Organic fertilizer selection and dosage recommendations Identified bacterial genera needs to be formulated for its survival so as to reach the soil for progress of crop productions. BioAtivo, Azotovit, Rhizotorphin, Azotoba cter in, Ekophit, Mizor in®, Ma mezo, Phylazonit-M, Symbion-N, CALOBIUM, Sardar Biofertilizers and Ferti-Bio are commonly available biofertilizers having identified genera in active state. Biofertilizers are usually bioformulated into two major type viz. liquid and solid with natural carriers. CONCLUSION This study implies the necessity of incorporating mathematical models to bring micro investigation on association of Insitu soil nutrient and its correlated bacterial genera so as to perceive the current nutrient status. Azotobacter, Enterobacter, Citrobacter, Achromobacter, E. coli, and Arthrobacter were some of the identified bacterial species that contribute to the soil primary macronutrients. This can be extempori- zing for organic fertilizer formulation consisting aforementioned identified bacterial genera and it is recommended to utilize those organic biofertlizers to attain massive yield in horticulture. ACKNOWLEDGEMENT We the authors grateful to support in the form of fellowship and encouragement received from SRM Institute of Science and Technology, Kattankulathur. It is pleasure to extend the acknowledgment to Dr. Akila and Vickram Muthu Rathinasabari for their valuable field assistance. Macronutrients and their associated bacterial genera J. Hortl. Sci. Vol. 17(1) : 131-136, 2022 136 REFERENCES Ar cha na , D. , Na ndish, M. , Sa va la gi, V. a nd Alaga wadi, A. 2013. Char acterization of potassium solubilizing bacteria (KSB) from rhizosphere soil. Q. J. Life Sci., 10:248-257. Aung, A., Sev, T.M., Mon, A.A. and Yu, S.S. 2020. Detection of abiotic stress tolerant Azotobacter species for enhancing plant growth promoting activities. 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Vol. 17(1) : 131-136, 2022 (Received: 22.10.2021; Revised: 17.02.2022; Accepted: 19.02.2022) 00 A Final SPH -JHS Coverpage First 2 pages.pdf 00 Content and in this issue.pdf 01 Mohan Kumar G N.pdf 02 Meera Pandey.pdf 03 Biradar C.pdf 04 Varalakshmi B.pdf 05 Vijayakumari N.pdf 06 Barik S.pdf 07 Sajid M B.pdf 08 Ranga D.pdf 09 Usha S.pdf 10 Manisha.pdf 11 Amulya R N.pdf 12 Akshatha H J.pdf 13 Adak T.pdf 14 Sujatha S.pdf 15 Gowda P P.pdf 16 Subba S.pdf 17 Dhayalan V.pdf 19 Ahmed S.pdf 20 Vishwakarma P K.pdf 21 Deep Lata.pdf 22 Udaykumar K P.pdf 23 Nayaka V S K.pdf 24 Sahel N A.pdf 25 Bayogan E R V.pdf 26 Rathinakumari A C.pdf 27 Yella Swami C.pdf 28 Saidulu Y.pdf 29 Sindhu S.pdf 30 Neeraj.pdf 31 Sivaranjani R.pdf 32 Rashied Tetteh.pdf 34 Sangeetha G.pdf 35 Shareefa M.pdf 36 Last Pages.pdf