Comparative Performance of the Microbial Supplements Voodoo Juice, Piranha Liquid, and Tarantula Liquid on Vegetative Growth and Flowering Product Efficacy Test Report Conducted by: Valentin Savov Konstantin Chakalov Todorka Popova Conducted at: Sofia University Saint Kliment Ohridski Faculty of Biology Department of Biotechnology 8 Dragan Tsankov Blvd. Sofia, Bulgaria Trial period: June 30 December 22, 2011
Sofia University Saint Kliment Ohridski 1 Introduction Hydroponic gardeners and horticulturalists use microbial supplements to bring the best of the soil food web its bottom tier, which consists of beneficial bacteria and fungi into their hydroponic gardens. In naturally rich soil, myriad beneficial microbes live symbiotically with plants, helping to strengthen their roots and increase their root mass. This relationship increases plants ability to absorb nutrients, thus directly contributing to greater vegetative growth and bigger yields. Beneficial microorganisms also help to keep the rhizosphere healthy by improving the microecological equilibrium, helping to fight off pathogens, and fulfilling many other advantageous functions (van der Heijden et al., 1998, Vessey 2003). However, beneficial microbes are typically absent from hydroponic growing systems and substrates, putting hydroponic root systems at a disadvantage. Therefore, certain hydroponic fertilizer manufacturers have taken to producing microbial supplements containing beneficial rhizobacteria and/ or rhizofungi in order to promote the colonization and inoculation of the rhizosphere by these naturally occurring, root-symbiotic microorganisms. Voodoo Juice, Piranha Liquid (Piranha), and Tarantula Liquid (Tarantula) are three microbial supplements produced by Advanced Nutrients, Ltd. (AN). Voodoo Juice and Tarantula contain beneficial bacterial species, while Piranha contains beneficial fungal and bacterial species. Theoretically, the application of such microbial supplements should result in numerous, significant benefits for plants grown hydroponically, including: Greater availability of nutrients due to microbes breaking them down into more bioavailable forms (Landeweert et al., 2001) and building up more root mass, resulting in higher absorption of nutrients and water (Vessey 2003). Beneficial microbes also secrete growth and bloom cofactors directly into plants, enabling them to increase growth and flowering (Gutiérrez-Mañero et al., 2001; Lebuhn, Heulin, and Hartmann, 1997; Strzelczyk and Pokojska-Burdziej, 1984; Timmusk et al., 1999). With these principles in mind, Voodoo Juice was developed to significantly expand plant root mass and therefore nutrient absorption. Biocontrol by means of mycoparasitism. Trichoderma, a powerful genus of fungi, is especially capable in this regard, devouring pathogenic fungi and bacteria and keeping the balance of microbial life in the rhizosphere in equilibrium. Trichoderma species have been widely used both as antagonistic fungal agents against several pests and as plant growth enhancers. Through mycoparasitism, they compete effectively for space and nutrients and kill pathogens through enzymes, antibiotics, and induced plant defense mechanisms (Verma et al., 2007). That way, no single microbial population is permitted to outnumber any other and create an imbalance. Piranha, which includes Trichoderma and other beneficial fungi, is intended to increase nutrient absorption and destroy harmful fungi and other pathogens, maintaining a healthy ecological balance among both beneficial and harmful microorganisms. A healthier rhizosphere due to the establishment of an ecological balance of rhizospheric microorganisms and better resistance to root diseases can also be achieved by beneficial bacteria. Many root diseases are caused by the decay of old roots and other organic waste matter, which can attract unwanted pathogens to the root zone. However, this waste can also be recycled by microbial enzymes into bioavailable forms. This recycling significantly increases the amount of available nutrients, such as nitrogen and phosphorus (Richardson et al., 2009). Several benefical bacterial species also increase the induced systemic resistance of plants and render them better able to fight off pathogens
Sofia University Saint Kliment Ohridski 2 (Kloepper, Ryu and Zhang, 2004). The species of beneficial bacteria contained in Tarantula are known to perform such functions. They are also intended to inoculate the roots, safeguarding them against diseases by competing with and warding off harmful bacteria and increasing the plant s systemic resistance to other pathogens. To view the species these three supplements contain, see Appendix II: Voodoo Juice, Piranha, and Tarantula labels. Based on our experience and on feedback from AN clients and customers, we have developed a hypothesis that Voodoo Juice, Piranha, and Tarantula are highly suitable for promoting flowering. Abundant anecdotal evidence suggests it is possible to significantly increase flowering using these products, even without the application of standard bloom-boosting supplements (i.e., those containing additional phosphorus and/or potassium augmenting the essential elements provided by the base fertilizer). Materials and methods Procedure The following products were obtained: 40 L bottles of ph P Grow, ph Micro, and ph P Bloom (ph P GMB) 40 L bottles of Voodoo Juice, Piranha, and Tarantula Nutrient solutions (NSes) were prepared using deionized RO water as follows: Background NS: ph P GMB according to the label application rate of 4 ml/l of ph Perfect Grow + 4 ml/l of ph Perfect Micro + 4 ml/l of Perfect Bloom NSes with microbial supplements: background NS + Voodoo Juice, Piranha, or Tarantula at 2 ml (i.e., equal to the label application rate) (Table 1) To test this hypothesis, we selected poinsettia, whose red-colored leaves, or bracts, develop by photoperiodism. Specifically, they require a sustained period (i.e., five or more days) of 12 hours of darkness to change color as well as abundant light during the day to achieve their richest coloration (Phipps, accessed in 2013). Therefore, the photoperiodic response of poinsettia resembles that of certain high-value crops frequently grown hydroponically. Variant Control Recommended dose of Voodoo Juice Recommended dose of Piranha Recommended dose of Tarantula NS ph P GMB: background Background + 2 ml/l of Voodoo Juice Background + 2 ml/l of Piranha Background + 2 ml/l of Tarantula The aim of this experiment was to determine the influence of Voodoo Juice, Piranha, and Tarantula when applied on top of base nutrients ph Perfect Grow (ph P Grow), ph Perfect Micro (ph P Micro), and ph Perfect Bloom (ph P Bloom) on the vegetative growth and flowering (i.e., red leaf development) of poinsettia, a short-day plant. Table 1. Test design for weeks 1 13. Well-developed Euphorbia pulcherrima (poinsettia cultivar Premium Red) cuttings, rooted and established quickly, were transplanted into 0.5 L pots containing the substrate Sunshine Mix #4 TM. Four plants were provided for each variant.
Sofia University Saint Kliment Ohridski 3 The irrigation was designed so that the solution could flow out. During daylight hours, one or two irrigations (using 0.07 0.08 L solution per plant for each irrigation) were done. The plants were grown in a phytochamber at 25 26 C during the daylight period (18 hours for the first four weeks (vegetative stage) and 10 hours for the next nine weeks the so-called bloom phase ) and at 18 20 C during the nighttime period. They were illuminated with 600 W/m 2 lamps. plants were irrigated with the same NS. Before each irrigation, the NSes were stirred vigorously so as to absorb gases from the atmosphere. Types of analysis Biometrical measurement of the weight of the fresh shoots and the dry shoots, the red leaf area, and the percentage of red leaf area to total leaf area. All NSes were prepared fresh every week. They were kept in bottles throughout the week and the same
Sofia University Saint Kliment Ohridski 4 Results and discussion The data represented in following charts show the results of the biometrical measurements. (To view the tables on which the charts are based, see Appendix I: Tables.) Total fresh weight of poinsettia shoots Percent vs. control; 2 ml 100.00 120.40 114.58 104.89 Control Voodoo Juice Piranha Tarantula Figure 1. Voodoo Juice, Piranha, and Tarantula increased fresh vegetative biomass by 20%, 15%, and 5%, respectively. Total dry weight of poinsettia shoots Percent vs. control; 2 ml 100.00 118.49 114.42 111.06 Control Voodoo Juice Piranha Tarantula Figure 2. Voodoo Juice, Piranha, and Tarantula increased dry vegetative biomass by 18%, 14%, and 11%, respectively.
Sofia University Saint Kliment Ohridski 5 Total red leaf area of poinsettias Percent vs. control; 2 ml 218.26 168.20 100.00 112.09 Control Voodoo Juice Piranha Tarantula Figure 3. Voodoo Juice, Piranha, and Tarantula increased yields (i.e., red leaf area) by 118%, 68%, and 12%, respectively. Total red leaf area of poinsettias Relative percent of red leaf area to total leaf area vs. control; 2 ml 19.79 39.82 32.30 25.77 Control Voodoo Juice Piranha Tarantula Figure 4. Voodoo Juice, Piranha, and Tarantula increased the percentage of red leaf area to total leaf area over the control by 40%, 32%, and 26%, respectively.
Sofia University Saint Kliment Ohridski 6 Vegetative growth All three supplements Voodoo Juice, Piranha, and Tarantula significantly influenced the formation of fresh shoots, increasing them by 20%, 15%, and 5%, respectively (Figure 1). Moreover, Voodoo Juice, Piranha, and Tarantula had a significant effect on dry-shoot weight, with Voodoo Juice increasing it by 18%, Piranha by 14%, and Tarantula by 11% (Figure 2). These positive results on vegetative growth were echoed in an experiment run with Piranha, Tarantula, and two competing microbial supplements on basil. Flowering As we had hypothesized, Voodoo Juice, Piranha, and Tarantula increased flowering (i.e., red leaf area) even without the application of standard bloom-boosting supplements. Although none of these supplements contain phosphorus, potassium, or other essential elements, all of them including Tarantula, which is intended to serve more as a biocontrol agent than as a nutrient-absorption enhancer had a direct positive impact on yields. as possible hormone-like activity of one or more species of beneficial bacteria. We expect Piranha s influence on flowering to be the result of increased root mass and more efficient nutrient absorption, as well as the beneficially antagonistic fungal activity of its fungal species which, like those in Tarantula, should serve as rhizospheric inoculants. We suspect Tarantula s more modest yet still significant increase in flowering was an indirect result of its intended specialization as a rhizospheric inoculant. Its Bacillus bacterial species are known to out-compete other pathogenic bacteria, ensuring a healthier, more ecologically balanced rhizosphere. This study demonstrated greater positive influence of Voodoo Juice, Piranha, and Tarantula on flowering than the control. These positive results on flowering were echoed in an experiment run with Voodoo Juice and two competing microbial supplements on chrysanthemum. Voodoo Juice significantly increased the percentage of red leaf area to the control indeed, by 118% while Piranha and Tarantula followed suit with a 68% and 12% yield increase, respectively (Figure 3). Voodoo Juice also significantly increased the ratio of red leaf to the total leaf area by 40%, Piranha by 32%, and Tarantula by 26% (Figure 4). In colloquial terms, this means all three microbial supplements produced much redder plants that is, plants characterized by more visible red leaf color than the control plants: We speculate that the impact of Voodoo Juice on flowering is the result of increased nutrient absorption due to a larger root system, as well Figure 5. Flowering poinsettias during the efficacy testing.
Sofia University Saint Kliment Ohridski 7 A few additional, interrelated comments are in order: 1. Taken together, these products contain numerous strains of bacteria and fungi that are often incompatible in concentrated solution and therefore need to be bottled separately to sustain viable populations. Once released in diluted form into the soil or into a nutrient reservoir of a hydroponic system, incompatibility should no longer pose a challenge. 2. Indeed, given that the action of these three products should theoretically complement each other and this has been substantiated by ample anecdotal evidence we postulate that excellent synergies should occur when Voodoo Juice, Piranha, and Tarantula are applied together. 3. Therefore, in order to confirm and determine the precise extent of these synergies, we plan to perform efficacy tests in which we will apply all three of these microbial supplements together. Since poinsettia (Figure 5) is a short-day plant, these findings could be of interest to gardeners and horticulturalists who are seeking to increase the flowering of other high-value short-day crops. * * * To learn more about the scientifically proven efficacy of Advanced Nutrients products and the science of hydroponics, download more white papers, efficacy reports, and special reports from the Hydroponics Research website at www.hydroponicsresearch.eu. Share this special report now with friends, coworkers, and family.
Sofia University Saint Kliment Ohridski 8 References Ames, R., Reid, C., Porter, L., and Cambardella, C., 1983. Hyphal uptake and transport of nitrogen from two 15 N-labelled sources by Glomus mosseae, a vesicular arbuscular mycorrhizal fungus. New Phytologist, 95 (3), pp. 381 396. Brownlee, C., Duddridge, J.A., Malibari, A., and Read, D.J., 1983. The structure and function of mycelial systems of ectomycorrhizal roots with special reference to their role in forming inter-plant connections and providing pathways for assimilate and water transport. Plant and Soil, 71, pp. 433 443. Gutiérrez-Mañero, F.J., et al., 2001. The plant-growth-promoting rhizobacteria Bacillus pumilus and Bacillus licheniformis produce high amounts of physiologically active gibberellins. Physiologia Plantarum, 111 (2), pp. 206 211. Kloepper, J.W., Ryu, C.M., and Zhang, S., 2004. Induced systemic resistance and promotion of plant growth by Bacillus spp. Phytopathology, 94 (11), pp. 1259 1266. Landeweert, R., et al., 2001. Linking plants to rocks: ectomycorrhizal fungi mobilize nutrients from minerals. Trends in Ecology & Evolution, 16 (5), pp. 248 254. Lebuhn, M., Heulin T., and Hartmann, A., 1997. Production of auxin and other indolic and phenolic compounds by Paenibacillus polymyxa strains isolated from different proximity to plant roots. FEMS Microbiology Ecology, 22 (4), pp. 325 334. Phipps, Nikki. How to make poinsettia turn red make a poinsettia rebloom, Gardening Know How, [online] Available at:<http://www.gardeningknowhow.com/ornamental/flowers/poinsettia/how-to-makepoinsettia-turn-red-make-a-poinsettia-rebloom.htm> [Accessed 5 August 2013]. Strzelczyk, E. and Pokojska-Burdziej, A., 1984. Production of auxins and gibberellin-like substances by mycorrhizal fungi, bacteria and actinomycetes isolated from soil and the mycorrhizosphere of pine (Pinus silvestris L.). Plant and Soil, 81 (2), pp. 185 194. Timmusk, S., Nicander, B., Granhall, U., and Tillberg, E., 1999. Cytokinin production by Paenibacillus polymyxa. Soil Biology & Biochemistry, 31 (13), pp. 1847 1852. van der Heijden, M.G., et al., 1998. Mycorrhizal fungal diversity determines plant biodiversity, ecosystem variability and productivity. Nature, 396, pp. 69 72. Verma, M., et al., 2007. Antagonistic fungi, Trichoderma spp: panoply of biological control. Biochemical Engineering Journal, 37 (1), pp. 1 20. Vessey, J.K. 2003. Plant growth promoting rhizobacteria as biofertilizers. Plant and Soil. 255, pp. 571 586.
Sofia University Saint Kliment Ohridski 9 Appendix I: Tables Voodoo Juice Control ph P GMB Recommended dose (2 ml/l) Fresh shoots, ave. g/plant 1st series 47.03 68.94 2nd series 100.31 108.46 Average 73.67 88.70 Dry shoots, ave. g/plant 1st series 14.30 15.37 2nd series 33.41 41.15 Average 23.85 28.26 Total green leaf area, cm 2 1st series 2,382.43 1,934.13 2nd series 2,023.83 1,650.67 Average 2,203.13 1,792.40 Total red leaf area, cm 2 1st series 261.53 1,180.51 2nd series 825.68 1,192.47 Average 543.61 1,186.49 % of red leaf area to total leaf area 1st series 9.89 37.90 2nd series 29.23 42.18 Average 19.56 40.04 Table 2. Biometric data of the poinsettia plants tested with Voodoo Juice.
Sofia University Saint Kliment Ohridski 10 Piranha Control ph P GMB Recommended dose (2 ml/l) Fresh shoots, ave. g/plant 1st series 47.03 63.74 2nd series 100.31 105.09 Average 73.67 84.41 Dry shoots, ave. g/plant 1st series 7.15 8.32 2nd series 16.70 18.98 Average 11.93 13.65 Total green leaf area, cm 2 1st series 2,382.43 1,745.33 2nd series 2,023.83 2,088.56 Average 2,203.13 1,916.95 Total red leaf area, cm 2 1st series 261.53 726.28 2nd series 825.68 1,102.38 Average 543.61 914.33 % of red leaf area to total leaf area 1st series 9.89 29.38 2nd series 29.23 34.77 Average 19.56 32.08 Table 3. Biometric data of the poinsettia plants tested with Piranha.
Sofia University Saint Kliment Ohridski 11 Tarantula Control ph P GMB Recommended dose (2 ml/l) Fresh shoots, ave. g/plant 1st series 47.03 56.59 2nd series 100.31 97.96 Average 73.67 77.27 Dry shoots, ave. g/plant 1st series 7.15 7.94 2nd series 16.70 18.55 Average 11.93 13.25 Total green leaf area, cm 2 1st series 2,382.43 1,770.53 2nd series 2,023.83 1,739.78 Average 2,203.13 1,755.15 Total red leaf area, cm 2 1st series 261.53 428.10 2nd series 825.68 790.54 Average 543.61 609.32 % of red leaf area to total leaf area 1st series 9.89 19.47 2nd series 29.23 31.12 Average 19.56 25.29 Table 4. Biometric data of the poinsettia plants tested with Tarantula.
Sofia University Saint Kliment Ohridski 12 Appendix: Voodoo Juice, Piranha, and Tarantula labels * 8 45268 00179 2 * The labels featured here were current at the time this efficacy report was published. Since that time, the labels may have been updated or amended. Therefore, the labels you see here may be obsolete. Do not print, copy, or distribute these labels.
Sofia University Saint Kliment Ohridski 13 8 45268 00033 7