Abstract
The application of chemical fertilizers to enhance crop production is a major concern due to associated environmental pollution and health hazards. Hence, there is an urgent need to develop an eco-friendly solution to improve crop production and promote sustainable agriculture simultaneously. Stevia rebaudiana is an important medicinal crop being substitute for sugar, superior flavor outline, extensive medicinal properties, and also of agronomic interest. In the present study, bacterium STJP isolated from the rhizospheric soil of S. rebaudiana and identified as Bacillus safensis on the basis of 16S rRNA gene sequencing, showed good amount of zinc (4.4 mg/L) and potassium (5.4 mg/L) solubilization. Paneer-whey (a dairy waste) based bioformulation (P-WBF) was developed utilizing isolate B. safensis STJP (accession number NAIMCC TB-2833) and inspected for the quality and ability to enhance the growth, nutrients uptake, and stevioside content in S. rebaudiana. The application of P-WBF displayed a significantly higher concentration (153.12%) of stevioside in S. rebaudiana as compared to control. P-WBF treated Stevia plants showed significantly higher fresh and dry weight as well (as compared to control). Further, enhancement of phosphorous, nitrogen, potassium, and zinc uptake in plant tissue was also recorded by application of P-WBF. This study suggests the use of P-WBF based biofertilizer using B. safensis STJP to increase stevioside content in Stevia plant by a nutrient(s) linked mechanism. This novel approach can also be beneficial for utilization of a dairy waste in preparation of bioformulation and, for enhancement of crop yield by an ecofriendly manner leading to sustainable agriculture.
Graphic abstract
![](http://media.springernature.com/lw685/springer-static/image/art%3A10.1007%2Fs11274-019-2783-x/MediaObjects/11274_2019_2783_Figa_HTML.png)
![](http://media.springernature.com/m312/springer-static/image/art%3A10.1007%2Fs11274-019-2783-x/MediaObjects/11274_2019_2783_Fig1_HTML.png)
![](http://media.springernature.com/m312/springer-static/image/art%3A10.1007%2Fs11274-019-2783-x/MediaObjects/11274_2019_2783_Fig2_HTML.png)
![](http://media.springernature.com/m312/springer-static/image/art%3A10.1007%2Fs11274-019-2783-x/MediaObjects/11274_2019_2783_Fig3_HTML.png)
![](http://media.springernature.com/m312/springer-static/image/art%3A10.1007%2Fs11274-019-2783-x/MediaObjects/11274_2019_2783_Fig4_HTML.png)
Similar content being viewed by others
References
Arora NK, Mishra J (2016) Prospecting the roles of metabolites and additives in future bioformulations for sustainable agriculture. App Soil Ecol 107:405–407. https://doi.org/10.1016/j.apsoil.2016.05.020
Arora NK, Kang SC, Maheshwari DK (2001) Isolation of siderophore-producing strains of Rhizobium meliloti and their biocontrol potential against Macrophomina phaseolina that causes charcoal rot of groundnut. Curr Sci 25:673–687
Arora NK, Khare E, Maheshwari DK (2010) Plant growth promoting rhizobacteria: constraints in bioformulation, commercialization, and future strategies. In: Plant growth and health promoting bacteria. Springer, Berlin. pp 97–116
Arora NK, Fatima T, Mishra I, Verma M, Mishra J, Mishra V (2018) Environmental sustainability: challenges and viable solutions. Environ Sustain 1(4):309–340. https://doi.org/10.1007/s42398-018-00038-w
Assaei R, Mokarram P, Dastghaib S, Darbandi S, Darbandi M, Zal F, Ranjbar Omrani GH (2016) Hypoglycemic effect of aquatic extract of stevia in pancreas of diabetic rats: PPARγ-dependent regulation or antioxidant potential. Avicenna J Med Biotechnol 8:65–74. https://doi.org/10.4172/2329-9029.1000158
Atkins CA, Smith PMC (2007) Translocation in legumes: assimilates, nutrients, and signaling molecules. Plant Physiol 144:550–561. https://doi.org/10.1104/pp.107.098046
Baba WN, Din S, Punoo HA, Wani TA, Ahmad M, Masoodi FA (2016) Comparison of cheese and paneer whey for production of a functional pineapple beverage: nutraceutical properties and Shelf life. J Food Sci Technol 53:2558–2568. https://doi.org/10.1007/s13197-016-2218-8
Berger LR, Stamford NP, Santos CERS, Freitas ADS, Franco LO, Stamford TCM (2013) Plant and soil characteristics affected by biofertilizers from rocks and organic matter inoculated with diazotrophic bacteria and fungi that produce chitosan. J Soil Sci Plant Nutr 13:592–603. https://doi.org/10.4067/s0718-95162013005000047
Biofertilizers and Organic Fertilizers in Fertilizer (Control) Order (1985) National Centre of Organic Farming Department of Agriculture and Cooperation, Ministry of Agriculture, Govt of India, CGO-II, Kamla Nehru Nagar Ghaziabad 201 001 Uttar Pradesh India India. https://ncof.dacnet.nic.in/training_manuals/training_manuals_in_english/bf_and_of_in_fco.pdf
Boonkaewwan C, Burodom A (2013) Anti-inflammatory and immunomodulatory activities of stevioside and steviol on colonic epithelial cells. J Sci Food Agric 93:3820–3825. https://doi.org/10.1002/jsfa.6287
Bovanová L, Brandšteterová E, Baxa S (1998) HPLC determination of stevioside in plant material and food samples. Zeitschrift für Lebensmitteluntersuchung und-Forschung A 207:352–365. https://doi.org/10.1007/s002170050344
Bräsen C, Esser D, Rauch B, Siebers B (2014) Carbohydrate metabolism in Archaea: current insights into unusual enzymes and pathways and their regulation. Microbiol Mol Biol Rev 78:89–175. https://doi.org/10.1128/mmbr.00041-13
Bureau of Indian Standards (2017) BIS act rules regulations https://bis.gov.in/?page_id=2377
Cantabella D, Piqueras A, Acosta-Motos JR, Bernal-Vicente A, Hernández JA, Díaz-Vivancos P (2017) Salt-tolerance mechanisms induced in Stevia rebaudiana Bertoni: Effects on mineral nutrition, antioxidative metabolism and steviol glycoside content. Plant Physiol Biochem 115:484–496. https://doi.org/10.1016/j.plaphy.2017.04.023
Chowdhary P, Yadav A, Singh R, Chandra R, Singh DP, Raj A, Bharagava RN (2018) Stress response of Triticum aestivum L. and Brassica juncea L. against heavy metals growing at distillery and tannery wastewater contaminated site. Chemosphere 206:122–131. https://doi.org/10.1016/j.chemosphere.2018.04.156
Das A, Biswas M, Mandal N (2010) An economic analysis of Stevia (Stevia rebaudiana Bert.) cultivation through stem cutting and tissue culture propagule in India. Trends Agric Econ 3:216–222. https://doi.org/10.3923/tae.2010.216.222
Debnath M, Ashwath N, Hill CB, Callahan DL, Dias DA, Jayasinghe NS, Midmore DJ, Roessner U (2018) Comparative metabolic and ionomic profiling of two cultivars of Stevia rebaudiana Bert. (Bertoni) grown under salinity stress. Plant Physiol Biochem 129:56–70. https://doi.org/10.1016/j.plaphy.2018.05.001
Dinesh R, Anandaraj M, Kumar A, Bini YK, Subila KP, Aravind R (2015) Isolation, characterization, and evaluation of multi-trait plant growth promoting rhizobacteria for their growth promoting and disease suppressing effects on ginger. Microbiol Res 173:34–43. https://doi.org/10.1016/j.micres.2015.01.014
Etesami H, Emami S, Alikhani HA (2017) Potassium solubilizing bacteria (KSB): Mechanisms, promotion of plant growth, and future prospects: a review. J Soil Sci Plant Nutr 17:897–911. https://doi.org/10.4067/s0718-95162017000400005
European Food Safety Authority (2010) Standard sample description for food and feed. EFSA J 8:1457. https://doi.org/10.2903/j.efsa.2010.1457
Fakhrul RH, Norrizah JS, Jaapar SS, Anilizawatima SN (2014) The effect of potassium concentrations on the growth and development of Stevia rebaudiana (Bertoni) and production of stevioside and rebaudioside A. Am-Eur J Sustain Agric 42–52
Food ro. Joint FAO/WHO Expert Committee on Food additives (JECFA) (2016) Eighty-third meeting Rome. 8–17 https://www.fao.org/3/a-bl776e.pdf
Fuwa K, Pulido P, McKay R, Vallee BL (1964) Determination of zinc in biological materials by atomic absorption spectrometry. Anal Chem 36:2407–2411. https://doi.org/10.1021/ac60219a009
Gaweł-Bęben K, Bujak T, Nizioł-Łukaszewska Z, Antosiewicz B, Jakubczyk A, Karaś M, Rybczyńska K (2015) Stevia rebaudiana Bert. leaf extracts as a multifunctional source of natural antioxidants. Molecules 20:5468–5486. https://doi.org/10.3390/molecules20045468
Goljanian-Tabrizi S, Amiri S, Nikaein D, Motesharrei Z (2016) The comparison of five low cost liquid formulations to preserve two phosphate solubilizing bacteria from the genera Pseudomonas and Pantoea. Iranian J Microbiol 8:377
Gomez KA, Gomez AA (1984) Statistical procedures for agricultural research. Wiley, New York
Gupta E, Kaushik S, Purwar S, Sharma R, Balapure AK, Sundaram S (2017) Anticancer potential of steviol in MCF-7 human breast cancer cells. Pharmacogn Magn 13:345. https://doi.org/10.4103/pm.pm_29_17
Hasanuzzaman M, Bhuyan MHMB, Nahar K, Hossain M, Mahmud J, Hossen M, Masud A, Fujita M (2018) Potassium: a vital regulator of plant responses and tolerance to abiotic stresses. Agronomy 8:31. https://doi.org/10.3390/agronomy8030031
Horwitz W, Chichilo P, Reynolds H (1970) Official methods of analysis of the association of official analytical chemists 10.1002/jps.2600600253
http://www.businessworld.in/article/The-Green-Sugar-Substitute/05-03-2018-142354/
http://www.krishisewa.com/articles/organic-agriculture/115-biofertilizers.html
Hu X, Chen J, Guo J (2006) Two phosphate-and potassium-solubilizing bacteria isolated from Tianmu Mountain, Zhejiang, China. World J Microbiol Biotechnol 22:983–990. https://doi.org/10.1007/s11274-006-9144-2
Jackson ML (1967) Soil Chemical Analysis Prentice Hall of India Pvt. Ltd. New Delhi 205. https://doi.org/10.1097/00010694-195806000-00014
Jayasinghe GY (2012) Sugarcane bagasses sewage sludge compost as a plant growth substrate and an option for waste management. Clean Technol Environ Policy 14:625–632. https://doi.org/10.1007/s10098-011-0423-8
Jimenez RR, Ladha JK (1993) Automated Elemental analysis—a rapid and reliable but expensive measurement of total carbon and nitrogen in plant and soil samples. Commun Soil Sci Plant Anal 24:1897–1924. https://doi.org/10.1080/00103629309368926
Jorjani M, Heydari A, Zamanizadeh HR, Rezaee S, Naraghi L (2011) Development of Pseudomonas fluorescens and Bacillus coagulans based bioformulations using organic and inorganic carriers and evaluation of their influence on growth parameters of sugar beet. J Biopest 4:180
Kalita M, Bharadwaz M, Dey T, Gogoi K, Dowarah P, Unni BG, Ozah D, Saikia I (2015) Develo** novel bacterial based bioformulation having PGPR properties for enhanced production of agricultural crops. Indian J Exp Bio 53:56–60
Kavamura VN, Santos SN, da Silva JL, Parma MM, Ávila LA, Visconti A, Zucchi TD, Taketani RG, Andreote FD, de Melo IS (2013) Screening of Brazilian cacti rhizobacteria for plant growth promotion under drought. Microbiol Res 168:183–191. https://doi.org/10.1016/j.micres.2012.12.002
Koenig R, Johnson C (1942) Colorimetric determination of phosphorus in biological materials. Ind Eng Chem Anal 14:155–166. https://doi.org/10.1021/i560102a026
Kregiel D (2015) Health safety of soft drinks: contents, containers, and microorganisms. BioMed Res Int. https://doi.org/10.1155/2015/128697
Kumar A, Singh M, Singh PP, Singh SK, Singh PK, Pandey KD (2016) Isolation of plant growth promoting rhizobacteria and their impact on growth and curcumin content in Curcuma longa L. Biocat Agri Biotechnol 8:1–7. https://doi.org/10.1016/j.bcab.2016.07.002
Lateef A, Adelere IA, Gueguim-Kana EB (2015) The biology and potential biotechnological applications of Bacillus safensis. Biologia 70:411–429. https://doi.org/10.1515/biolog-2015-0062
Latha S, Chaudhary S, Ray RS (2017) Hydroalcoholic extract of Stevia rebaudiana bert. leaves and stevioside ameliorates lipopolysaccharide induced acute liver injury in rats. Biomed Pharmacother 95:1040–1050. https://doi.org/10.1016/j.biopha.2017.08.082
Lemus-Mondaca R, Vega-Gálvez A, Zura-Bravo L, Ah-Hen K (2012) Stevia rebaudiana Bertoni, source of a high-potency natural sweetener: a comprehensive review on the biochemical, nutritional and functional aspects. Food chem 132:1121–1132. https://doi.org/10.1016/j.foodchem.2011.11.140
Liu X, Ren G, Shi Y (2011) The effect of organic manure and chemical fertilizer on growth and development of Stevia rebaudiana Bertoni. Energy Procedia 5:1200–1214. https://doi.org/10.1016/j.egypro.2011.03.210
Mahanty T, Bhattacharjee S, Goswami M, Bhattacharyya P, Das B, Ghosh A, Tribedi P (2017) Biofertilizers: a potential approach for sustainable agriculture development. Environ Sci Pollut Res 24:3315–3335. https://doi.org/10.1007/s11356-016-8104-0
Mamta RP, Pathania V, Gulati A, Singh B, Bhanwra RK, Tewari R (2010) Stimulatory effect of phosphate-solubilizing bacteria on plant growth, stevioside and rebaudioside-A contents of Stevia rebaudiana Bertoni. Appl Soil Ecol 46:222–239. https://doi.org/10.1016/j.apsoil.2010.08.008
Masclaux-Daubresse C, Daniel-Vedele F, Dechorgnat J, Chardon F, Gaufichon L, Suzuki A (2010) Nitrogen uptake, assimilation and remobilization in plants: challenges for sustainable and productive agriculture. Ann Bot 105:1141–1157
Mayer FL, Kronstad JW (2017) Disarming fungal pathogens: Bacillus safensis inhibits virulence factor production and biofilm formation by Cryptococcus neoformans and Candida albicans. MBio Kavamura 8:01537–1617. https://doi.org/10.1128/mbio.01537-17
Megeji NW, Kumar JK, Singh V, Kaul VK, Ahuja PS (2005) Introducing Stevia rebaudiana, a natural zero-calorie sweetener. Curr Sci 10:801–814
Mishra J, Prakash J, Arora NK (2016) Role of beneficial soil microbes in sustainable agriculture and environmental management. Cli Chang Environ Sustain 4:137–149. https://doi.org/10.5958/2320-642x.2016.00015.6
Modi A, Litoriya N, Prajapati V, Rafalia R, Narayanan S (2014) Transcriptional profiling of genes involved in steviol glycoside biosynthesis in Stevia rebaudiana bertoni during plant hardening. Dev Dyn 243:1067–1073. https://doi.org/10.1002/dvdy.24157
Mukherjee P, Roychowdhury R, Roy M (2017) Phytoremediation potential of rhizobacterial isolates from Kans grass (Saccharum spontaneum) of fly ash ponds. Clean Techn Environ Policy 19:1373–1385. https://doi.org/10.1007/s10098-017-1336-y
Mumtaz MZ, Ahmad M, Jamil M, Hussain T (2017) Zinc solubilizing Bacillus spp. potential candidates for biofortification in maize. Microbiol res 202:51–60. https://doi.org/10.1016/j.micres.2017.06.001
Nagachandrabose S (2018) Liquid bioformulations for the management of root-knot nematode, Meloidogyne hapla that infects carrot. Crop Prot 114:155–161. https://doi.org/10.1016/j.cropro.2018.08.022
Nieves-Cordones MA, Shiblawi FR, Sentenac H (2016) Roles and transport of sodium and potassium in plants. InThe Alkali Metal Ions: Their Role for Life. Springer Cham 291–324
Page AL, Miller RH, Keeney DR (1982) Methods of soil analysis; 2. Chemical and microbiological properties, 2. Aufl. 1184 S., American Soc. of Agronomy (Publ.), Madison, Wisconsin, USA, gebunden 36 Dollar. Zeitschrift für Pflanzenernährung und Bodenkunde. 148:363–374. https://doi.org/10.1002/jpln.19851480319
Pal PK, Kumar R, Guleria V, Mahajan M, Prasad R, Pathania V, Gill BS, Singh D, Chand G, Singh B, Singh RD (2015) Crop-ecology and nutritional variability influence growth and secondary metabolites of Stevia rebaudiana Bertoni. BMC Plant Biol 15:67. https://doi.org/10.1186/s12870-015-0457-x
Pandey P, Maheshwari DK (2007) Bioformulation of Burkholderia sp. MSSP with a multispecies consortium for growth promotion of Cajanus cajan. Can J Microbiol 53:213–222. https://doi.org/10.1139/w06-118
Panghal A, Kumar V, Dhull SB, Gat Y, Chhikara N (2017) Utilization of dairy industry waste-whey in formulation of papaya RTS beverage. Curr Res Nutr Food Sci J 20:168–174. https://doi.org/10.12944/crnfsj.5.2.14
Parmar P, Sindhu SS (2013) Potassium solubilization by rhizosphere bacteria: influence of nutritional and environmental conditions. J Microbiol Res 3:25–31
Pirlak L, Turan M, Sahin F, Esitken A (2007) Floral and foliar application of plant growth promoting rhizobacteria (PGPR) to apples increases yield, growth, and nutrient element contents of leaves. J Sustain Agric 30:145–155. https://doi.org/10.1300/j064v30n04_11
Prakash J and Arora NK (2019) Phosphate-solubilizing Bacillus sp. enhances growth, phosphorus uptake and oil yield of Mentha arvensis L. Biotech (2019) 9:126
Rai C, Majumdar GC, De S (2012) Optimization of process parameters for water extraction of stevioside using response surface methodology. Sep Sci Technol 47:1014–1022. https://doi.org/10.1080/01496395.2011.641055
Richman AS, Gijzen M, Starratt AN, Yang Z, Brandle JE (1999) Diterpene synthesis in Stevia rebaudiana: recruitment and up-regulation of key enzymes from the gibberellin biosynthetic pathway. Plant J 19:411–421. https://doi.org/10.1046/j.1365-313x.1999.00531.x
Salazar VAG, Encalada SV, Cruz AC, Campos MRS (2018) Stevia rebaudiana: a sweetener and potential bioactive ingredient in the development of functional cookies. J Funct Foods 44:183–190. https://doi.org/10.1016/j.jff.2018.03.007
Saravanan VS, Subramoniam SR, Raj SA (2004) Assessing in vitro solubilization potential of different zinc solubilizing bacterial (ZSB) isolates. Braz J Microbiol 35:121–135. https://doi.org/10.1590/s1517-83822004000100020
Savci S (2012) Investigation of effect of chemical fertilizers on environment. Apcbee Procedia 1:287–292. https://doi.org/10.1016/j.apcbee.2012.03.047
Shakeel M, Rais A, Hassan MN, Hafeez FY (2015) Root associated Bacillus sp. improves growth, yield and zinc translocation for basmati rice (Oryza sativa) varieties. Front Microbiol 18:1286
Sheirdil RA, Hayat R, Zhang XX, Abbasi NA, Ali S, Ahmed M, Khattak JZ, Ahmad S (2019) Exploring potential soil bacteria for sustainable wheat (Triticum aestivum L.) production. Sustainability 11(12):3361
Šic Žlabur J, Voća S, Dobričević N, Ježek D, Bosiljkov T, Brnčić M (2013) Stevia rebaudiana Bertoni—a review of nutritional and biochemical properties of natural sweetener. Agric Consp Sci 78:25–30
Smith-Hall C, Larsen HO, Pouliot M (2012) People, plants and health: a conceptual framework for assessing changes in medicinal plant consumption. J Ethnobiol Ethnomed 8:43. https://doi.org/10.1186/1746-4269-8-43
Tandel KR (2011) Sugar substitutes: Health controversy over perceived benefits. J Pharmacol Pharmacotherap 2:236. https://doi.org/10.4103/0976-500x.85936
Tanih NF, Sekwadi E, Ndip RN, Bessong PO (2015) Detection of pathogenic Escherichia coli and Staphylococcus aureus from cattle and pigs slaughtered in abattoirs in Vhembe District South Africa. Sci World J 20:15. https://doi.org/10.1155/2015/195972
Teixeira WF, Fagan EB, Soares LH, Umburanas RC, Reichardt K, Neto DD (2017) Foliar and seed application of amino acids affects the antioxidant metabolism of the soybean crop. Front Plant Sci 8:327. https://doi.org/10.3389/fpls.2017.00327
Wang M, Zheng Q, Shen Q, Guo S (2013) The critical role of potassium in plant stress response. Inter J Mol Sci 14:7370–7390. https://doi.org/10.3390/ijms14047370
Whalen JK (2014) Managing soil biota-mediated decomposition and nutrient mineralization in sustainable agroecosystems. Adv Agri. https://doi.org/10.1155/2014/384604
Yadav AK, Singh S, Dhyani D, Ahuja PS (2011) A review on the improvement of stevia [Stevia rebaudiana (Bertoni)]. Can J Plant Sci 91:1–27. https://doi.org/10.4141/cjps10086
Zaman MM, Rahman MA, Chowdhury T, Chowdhury MAH (2018) Effects of combined application of chemical fertilizer and vermicompost on soil fertility, leaf yield and stevioside content of stevia. J Bangl Agric Univ 16:73–81. https://doi.org/10.3329/jbau.v16i1.36484
Acknowledgements
The authors was highly obliged to Vice chancellor of BBA University (A Central University) providing facility for all experimental work. Authors are also grateful to university science instrument centre (USIC) for providing HPLC facility.
Author information
Authors and Affiliations
Corresponding author
Ethics declarations
Conflict of interest
All authors declare that there is no conflict of interest in this original article.
Additional information
Publisher's Note
Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.
Electronic supplementary material
Below is the link to the electronic supplementary material.
Rights and permissions
About this article
Cite this article
Prakash, J., Arora, N.K. Development of Bacillus safensis-based liquid bioformulation to augment growth, stevioside content, and nutrient uptake in Stevia rebaudiana. World J Microbiol Biotechnol 36, 8 (2020). https://doi.org/10.1007/s11274-019-2783-x
Received:
Accepted:
Published:
DOI: https://doi.org/10.1007/s11274-019-2783-x