Int J Recycl Org Waste Agricult (2015) 4:279–289 DOI 10.1007/s40093-015-0107-1 ORIGINAL RESEARCH Cultivable bacterial diversity and early plant growth promotion by the traditional organic formulations prepared using organic waste materials 1,2 1 2 Rangasamy Anandham • Nagaiah Premalatha • Hyeong Jin Jee • 3 3 1 Hang Yeon Weon • Soon Wo Kwon • Ramasamy Krishnamoorthy • 4 2 1 Pandiyan Indira Gandhi • Yong Ki Kim • Nellaiappan Olaganathan Gopal Received: 2 June 2015 / Accepted: 5 October 2015 / Published online: 9 November 2015 Ó The Author(s) 2015. This article is published with open access at Springerlink.com Abstract Results Bacterial 16SrDNA analysis revealed that the Purpose Traditional organic formulations are widely isolates fell into forty-three different genera, which can be used as a plant growth promoters; however, the knowledge grouped into seven different classes, such as Alphapro- on the microbial aspect of traditional organic formulations teobacteria, Actinobacteria, Bacilli (or Firmibacteria), is still limited. The aim of this study was to illustrate the Betaproteobacteria, Cytophagia, Flavobacteriia and cultivable bacterial diversity of various traditional organic Gammaproteobacteria. Higher bacterial diversity was formulations and their potential for early plant growth observed in cow dung followed by 33 and 100 % pan- promotion. chagavya. Radish and Chinese cabbage seed germination Methods Five different traditional organic formulations and growth were significantly improved by traditional such as 100 % panchagavya,33%panchagavya, plant organic formulations compared to control. extract with native microorganisms, commercial organic Conclusion The results of this study showed that the fertilizer extract with two percent leaf soil extract and bacterial diversity changes depend on the type and con- commercial organic fertilizer extract with 2 % yogurt were centration of ingredients used in traditional organic for- prepared and used in this study. The liquid fraction of these mulations. Substantial increase in plant growth by the traditional organic formulations were used to analyze the traditional organic formulations indicates the suitability of beneficial effect on plant growth by seed treatment and using these organic preparations in eco-friendly foliar applications. agriculture. Keywords Eubacterial diversity Á Organic farming Á Electronic supplementary material The online version of this article (doi:10.1007/s40093-015-0107-1) contains supplementary Organic formulations Á Panchagavya Á Plant growth- material, which is available to authorized users. promoting bacteria & Rangasamy Anandham [email protected] of SID Introduction 1 Department of Agricultural Microbiology, Agricultural College and Research Institute, Tamil Nadu Agricultural Intensive agriculture depends on expensive chemical inputs University, Madurai 625 104, Tamil Nadu, India from off farm, which generates lot of pollutants and affects 2 Organic Agriculture Division, National Academy of environment (Horrigan et al. 2002). Due to the negative Agricultural Science, Rural Development Administration, impact of chemical inputs, organic farming gained more Suwon 441-707, Republic of Korea 3 attention from last few decades (Badgley et al. 2007). The Korean Agricultural Culture Collection, National Agro Food and Agricultural Organization of the United Nations Biodiversity Center, National Academy of Agricultural Science, Rural Development Administration, (FAO) estimated that the area under organic agriculture Suwon 441-707, Republic of Korea was 37.2 million ha in 2011, which was three times higher 4 Regional Research Station, Tamil Nadu Agricultural than that of 1999 (FAO 2013). In organic farming, syn- University, Vridhachalam 606 001, Tamil Nadu, India thetic pesticides, mineral fertilizers, genetically modified www.SID.ir123 280 Int J Recycl Org Waste Agricult (2015) 4:279–289 organisms and sewage sludge applications are strictly applications such as developments of antibiotics or excluded. Nonetheless, biofertilizers or traditional organic biofertilizers. Only a few reports elucidate the microbial formulations prepared by using organic materials/wastes population in different organic formulations prepared by are encouraged to use as a plant nutrition (Badgley et al. farmers (Radha and Rao 2014; Amalraj et al. 2013; Girija 2007). Numerous studies stated that the yield and quality of et al. 2013). Recently, higher number of cultivable bacte- plant products produced by organic farming are consider- rial genera was obtained from the organic formulation ably better than that of conventional farming (Chemura prepared using fermented cow manure (Giannattasio et al. 2014; Oliveira et al. 2013; Luthria et al. 2010). Organic 2013). In addition, few novel and plant growth-promoting farming not only improves the quality of the food products, bacteria such as Larkinella bovis sp. nov. and Microbac- but it also improves the soil fertility. In a long-term study terium suwonense sp. nov. were isolated from traditional in Switzerland, organic farming with composted animal organic formulations and tested for their plant growth manure improved soil quality significantly as compared to promotion (Anandham et al. 2011a, b; Sreenivasa et al. conventional farming with mineral fertilizer (Maeder et al. 2009). 2002). In this study, culture-dependent method was used to Traditional organic formulations may contain numerous illustrate the bacterial diversity in various traditional plant growth-promoting bacteria (PGPB), which may organic formulations made from organic materials and enhance plant growth by nitrogen fixation, growth hormone evaluated the effect of organic formulations on early production and control phytopathogens (Amalraj et al. growth of radish and Chinese cabbage. Successfully culti- 2013; Naik and Sreenivasa 2009). In many Asian countries, vated bacterial isolates may be used to develop a microbial farmers formulate their own organic formulations by consortium to support sustainable agricultural practice in combining different organic materials and treating them by future. fermentation or composting. For example, in India, pan- chagavya (PG) is one of the widely used traditional organic formulations, which is mostly prepared by farmers them- Materials and methods selves. Panchagavya is a fermented product made from five ingredients obtained from cow, such as milk, urine, Traditional organic formulations preparations dung, curd and clarified butter (Amalraj et al. 2013). South Korean farmers use wild weed wineberry (Rubus phoeni- Organic formulations such as 100 % PG, 33 % PG, plant colasius) extract with native microorganisms, which is extract with native microorganism (PNM), commercial prepared by mixing wild weed extract, salt, molasses and organic fertilizer extract with 2 % leaf soil extract (OLE) decomposed leaf soil extract. Thailand farmers use fer- and commercial organic fertilizer extract with 2 % yogurt mented plant extracts or fermented plant juices, which can (OY) were prepared and used in this study. Ingredients and promote plant growth and act as a biocontrol agent (Kan- their concentration of different traditional organic formu- tachote et al. 2009). Bokashi is a Japanese traditional lations are given in Table 1. organic formulation preparation which contains effective Commercially available pelleted organic fertilizer (OF) microorganisms (EM). Japanese first developed the EM extract was used to prepare OLE and OY. Pelleted OF concept during 1980s, and this preparation contains various contains 55 % caster bean waste, 40 % rice bran and 5 % PGPB and yeast (Yamada and Xu 2001). These traditional palm waste with the nutrient content of 45 % N, 1.5 % P organic formulations have not been well characterized and 1.0 % K. For preparing an organic fertilizer extract, microbiologically. forty gram OF was boiled in 400 ml of water (sterile dis- In recent past, severalArchive culture-independent methods tilled of water) for 30SID min and then the filtrate was collected have been developed to illustrate higher percent of bacte- using the filter paper (Whatman No. 2–90 mm). Leaf soil rial population than usually found with cultural-dependent extract was prepared by mixing 20 g of soil [containing methods (Videira et al. 2013). These culture-independent decomposed leaf, collected from the hilly region Gangwon methods may give in-depth knowledge about bacterial Province of South Korea (37°49022.6400N; diversity, but it is impossible to use this information to 128°09023.0000E)] with 120 ml of water (sterile distilled identify the physiology of particular bacterial strain, water) and shaken for 30 min, then filtered through filter interaction between microorganisms and response of bac- paper (Whatman No. 2–90 mm). The ingredients of the terial strains to different environmental conditions (Stewart each traditional organic formulation were mixed in a flask 2012). However, the culture-dependent method can be and allowed for fermentation for 5 days in shaking con- more useful in studying bacterial physiology and their role dition (120 rpm) at 30 °C. During fermentation, microor- in plant growth promotion (Palleroni 1997). In addition, a ganisms can degrade the organic materials and enhance the cultural-dependent method may be helpful in industrial production of organic formulations. All five organic 123 www.SID.ir Int J Recycl Org Waste Agricult (2015) 4:279–289 281 Table 1 Ingredients and their concentrations of different organic formulations 100 % PG 33