THE PROOF FILES: VOODOO JUICE RESEARCH DOSSIER
© 2008 Advanced Nutrients Research
International Hydroponics Research Team
Voodoo Juice beneficial bacteria creates root mass growth:
The bacteria penetrate the cortex, induce root nodules, multiply and subsequently differentiate into bacteroids, which produce the nitrogenase enzyme complex. Within the root nodules, the plant creates a low oxygen concentration, which allows bacterial nitrogenase to convert atmospheric nitrogen into ammonia. In return, the plant supplies the bacteria with a carbon source.
“Molecular basis of plant growth promotion and biocontrol by rhizobacteria” in Current Opinion in Plant Biology. 2001. 4(4):343 – 350.
- Penetration results in nodules, nodules result in increased root surface area bacteria at the root tip are metabolically most active
“Molecular basis of plant growth promotion and biocontrol by rhizobacteria” in Current Opinion in Plant Biology. 2001. 4(4):343 – 350.
- Lots of growth going on stimulates growth of bacteria, which in turn stimulate further plant growth.
HPLC analysis confirmed the presence of indole acetic acid (IAA) and indole acetamide (IAM) as the major auxins in the culture filtrates of these rhizobacteria. A series of laboratory experiments conducted on two cv. of wheat under gnotobiotic (axenic) conditions demonstrated increases in root elongation (up to 17•3%), root dry weight (up to 13•5%), shoot elongation (up to 37•7%) and shoot dry weight (up to 36•3%) of inoculated wheat seedlings.
“Screening plant growth-promoting rhizobacteria for improving growth and yield of wheat” in Journal of Applied Microbiology. 2004. 96 (3): 473–480.
There was significant linear correlation between auxins produced by rhizobacteria in vitro and growth of wheat seedlings (particularly root and shoot weights) under gnotobiotic conditions. This may imply that auxins produced by PGPR isolates caused improvement in root system, resulting in more biomass production.
“Screening plant growth-promoting rhizobacteria for improving growth and yield of wheat” in Journal of Applied Microbiology. 2004. 96 (3): 473–480.
inoculated plants resulted in better germination, early development and flowering and an increase in dry weight of both the root system and the upper plant parts. There was a positive correlation between the increase in yield and the improvement of root development. Similarly, promotion in plant height, number of tillers, plant dry weight and grain yields of various crop plants in response to inoculation with PGPR were reported by other workers.
"Screening plant growth-promoting rhizobacteria for improving growth and yield of wheat" in Journal of Applied Microbiology. 2004. 96 (3): 473–480.
By far the most evidence for the positive effects of biofertilizing-PGPR points to bacteria-mediated changes in root growth and morphology. Bacterial mediated increases in root weight are commonly reported responses to PGPR inoculations (e.g., Bashan and Dubrovsky, 1996; Bertrand et al., 2001; Frommel et al., 1991; Vessey and Buss, 2002). More importantly, increases in root length and root surface area are sometimes reported.
“Plant growth promoting rhizobacteria as biofertilizers” in Plant and Soil. 2003. 255: 571–586.
The reporting of root length and root surface area are important because increase in these parameters are more reflective of an increase the volume of soil explored, than that which would be indicated by just increases in root weight.
“Plant growth promoting rhizobacteria as biofertilizers” in Plant and Soil. 2003. 255: 571–586.
Indole-3-acetic acid is a phytohormone that is known to be involved in root initiation, cell division, and cell enlargement (Salisbury, 1994). This hormone is very commonly produced by PGPR (Barazani and Friedman, 1999).
“Plant growth promoting rhizobacteria as biofertilizers” in Plant and Soil. 2003. 255: 571–586.
Most commonly, IAA-producing PGPR are believed to increase root growth and root length, resulting in greater root surface area that enables the plant to access more nutrients from soil.
“Plant growth promoting rhizobacteria as biofertilizers” in Plant and Soil. 2003. 255: 571–586.
PGPR are beneficial for plant growth and also referred as yield increasing bacteria (YIB). They can affect plant growth and yield in a number of ways and enhancement of vegetative and reproductive growth is documented in a range of crops like cereals, pulses, ornamentals, vegetables, plantation crops and some trees. Treatments with PGPR increase germination percentage, seedling vigor, emergence, plant stand, root and shoot growth, total biomass of the plants, seed weight, early flowering, grains, fodder and fruit yields etc., (van Loon et al., 1998; Ramamoorthy et al., 2001).
Siddiqui, Z.A. (ed.). PGPR: Biocontrol and Biofertilization. Springer. 2005. p197-216
The application of five commercial chitosan-based formulations of carefully chosen PGPR developed at Auburn University, USA has previously shown demonstrable increase in the growth of nursery-raised plants such as cucumber, pepper and tomato among others. Later, seedlings of three indica rice cultivars, IR24, IP50 and Jyothi raised in rice field soil amended with each of the formulations in a 1:40 (formulation: soil) ratio have shown significant two-fold increase in root and shoot length, and grain yield. The observations do suggest that application of such commercial bacterial formulations can serve as microbial inoculants for the improvement of rice growth (Vasudevan et al., 2002).
Siddiqui, Z.A. (ed.). PGPR: Biocontrol and Biofertilization. Springer. 2005. p197-216
The overproduction of ethylene in response to abiotic and biotic stresses leads to inhibition of root growth and consequently growth of the plant as a whole. Ethylene synthesis is stimulated by a variety of environmental factors/stresses, which hamper plant growth [1]. As described earlier, PGPR containing ACC deaminase regulate and lower the levels of ethylene by metabolizing ACC; a precursor of plant produced ethylene. These ACC deaminase PGPR boost plant growth particularly under stressed conditions by the regulation of accelerated ethylene production in response to a multitude of abiotic and biotic stresses like salinity, drought, waterlogging, temperature, pathogenicity and contaminants.
“Perspective of plant growth promoting rhizobacteria (PGPR) containing ACC deaminase in stress agriculture.” Journal of Industrial Microbiology and Biotechnology. 2007. 34:635–648
Inoculation of European alder seedlings with bacterial strain B. licheniformis grown in Soil A resulted in a significant increase in root and leaf surface area over non-inoculated controls. Differences in root surface area and nodulation between treated and non-treated seedlings were observed at 6 wai and were twice that of non-inoculated seedlings by 8 wai. However, shoot and root lengths were unaffected by bacteria at the end of the experiment.
“Alterations in the rhizobacterial community associated with European alder growth when inoculated with PGPR strain Bacillus licheniformis” Environmental and Experimental Botany. 2003. 49(1): 61-68
When alder seedlings grown in Soil B were inoculated, no differences were observed in the above ground portion although B. licheniformis significantly increased both root length and surface area. Similarly to seedlings from Soil A, root surface was significantly affected and inoculated seedlings showed 2-fold increases in root surface compared with controls. No nodules were detected in this soil along the experiment.
“Alterations in the rhizobacterial community associated with European alder growth when inoculated with PGPR strain Bacillus licheniformis” Environmental and Experimental Botany. 2003. 49(1): 61-68
Production of auxin-like compounds could explain two things: (i) differences in the nodulation process in which auxins have been involved ( Frankenberger and Arshad, 1995), and (ii) increases detected in root parametres ( Selvadurai et al., 1991), since auxins affect root growth as well as root growth pattern, increasing root-soil surface, therefore, improving nutrient and water absorption potential ( Germida and Walley, 1996).
“Alterations in the rhizobacterial community associated with European alder growth when inoculated with PGPR strain Bacillus licheniformis” Environmental and Experimental Botany. 2003. 49(1): 61-68
- How PGPR make root masses larger, heavier, or more extensive is not well known. It is thought partly to be due to hormones produced by the bacteria or simply the colonization process itself, as these bacteria live INSIDE and around the rest of the ball, producing bumpy outgrowths (nodes) that increase root mass, volume, and surface area
Voodoo Juice Promotes Plant Growth
Bacillus spp.
In vivo, eight PGPR strains (Bacillus subtilis GB03, B. amyloliquefaciens IN937a, B. pumilus SE-34, B. pumilus T4, B. pasteurii C9, Paenibacillus polymyxa E681, Pseudomonas fluorescens 89B-61, and Serratia marcescens 90-166) promoted foliar fresh weight under greenhouse conditions 4 weeks after sowing… Elicitation of growth promotion by PGPR strains in vitro involved signaling of brassinosteroid, IAA, salicylic acid, and gibberellins.
Study of mechanisms for plant growth promotion elicited by rhizobacteria in Arabidopsis thaliana. 2005. Plant and Soil. 268(1): 285-292.
Bacillus spp
The demonstration that PGPR strains release different volatile blends and that plant growth is stimulated by differences in these volatile blends establishes an additional function for volatile organic compounds as signaling molecules mediating plant–microbe interactions.
Bacterial volatiles promote growth in Arabidopsis. 2003. PNAS 100 (8): 4927–4932.
Bacillus spp
The production of phytohormones, namely auxin, cytokinin and gibberellin are the most commonly invoked mechanisms of plant growth promotion by PGPR.
Intracellular and extracellular PGPR: commonalities and distinctions in the plant–bacterium signaling processes. 2005. Soil Biology and Biochemistry. 37(3): 395-412.
Bacillus spp.
[Bacillus spp. produce] high levels of gibberellins: A1, A3, A4 and A20, inducing positive Bacillus spp effects on stem and shoot elongation.
Intracellular and extracellular PGPR: commonalities and distinctions in the plant–bacterium signaling processes. 2005. Soil Biology and Biochemistry. 37(3): 395-412.
Bacillus spp.
In most cases, Bacillus spp. that elicit ISR also elicit plant growth promotion.
“Induced Systemic Resistance and Promotion of Plant Growth by Bacillus spp.” In Phytopathology. 2004. 94(11): 1259-1266.
Bacillus spp.
The mixture of strains IN937a and IN937b also resulted in significant yield increases of all crops during the rainy season and generally in significant promotion of midseason plant growth.
“Induced Systemic Resistance and Promotion of Plant Growth by Bacillus spp.” In Phytopathology. 2004. 94(11): 1259-1266.
Bacillus spp.
VOCs from two different Bacillus spp., which are known elicitors of ISR, elicit plant growth promotion and ISR
“Induced Systemic Resistance and Promotion of Plant Growth by Bacillus spp.” In Phytopathology. 2004. 94(11): 1259-1266.
- VOC = volatile organic compound
Bacillus spp.
Numerous species of soil bacteria which flourish in the rhizosphere of plants, but which may grow in, on, or around plant tissues, stimulate plant growth by a plethora of mechanisms. These bacteria are collectively known as PGPR (plant growth promoting rhizobacteria).
“Plant growth promoting rhizobacteria as biofertilizers” in Plant and Soil. 2003. 255(2): 1573-5036.
Bacillus spp.
The means by which PGPR enhance the nutrient status of host plants can be categorized into five areas: (1) Biological N2 fixation, (2) increasing the availability of nutrients in the rhizosphere, (3) inducing increases in root surface area, (4) enhancing other beneficial symbioses of the host, and (5) combination of modes of action.
“Plant growth promoting rhizobacteria as biofertilizers” in Plant and Soil. 2003. 255(2): 1573-5036.
Protects against plant pathogens and disease
Bacillus licheniformis
Bacillus licheniformis producing an antifungal principle active against a wide variety of plant pathogenic fungi has been isolated…The antifungal principle is fungistatic against several diverse species of fungi, and shows particularly strong activity against filamentous fungi, many of which are serious plant pathogens.
Bacillus licheniformis producing antifungal agents and uses thereof for control of phytopathogenic fungi. United States Patent 5665354.
Bacillus licheniformis
Bacillus licheniformis is widely known to produce antibiotics such as bacillomycin, bacitracin, licheniformin and proticin (See Katz & Demain (1977) , Bacteriol. Rev. 41: 449-474).
Bacillus Licheniformis Producing Antifungal Agents and Uses thereof for Control of Phytopathogenic Fungi. US Patent PCT/US1995/008536.
Bacillus licheniformis
Bacillus licheniformis is antagonistic to a variety of plant pathogenic fungi, including several common pathogens of turf grass and other graminaceous crops, such as corn (Zea mays L.) …Bacillus licheniformis appears to exert its antagonistic effect by producing one or more substances having antifungal activity.
Bacillus Licheniformis Producing Antifungal Agents and Uses thereof for Control of Phytopathogenic Fungi. US Pantent PCT/US1995/008536.
Bacillus spp.
Published results are summarized showing that specific strains of the species B. amyloliquefaciens, B. subtilis, B. pasteurii, B. cereus, B. pumilus, B. mycoides, and B. sphaericus elicit significant reductions in the incidence or severity of various diseases on a diversity of hosts.
“Induced Systemic Resistance and Promotion of Plant Growth by Bacillus spp.” In Phytopathology. 2004. 94(11): 1259-1266.
Bacillus spp.
Protection resulting from [induced systematic resistance] elicited by Bacillus spp. (B. amyloliquefaciens, B. subtilis, B. pasteurii, B. cereus, B. pumilus, B. mycoides, and B. sphaericus) has been reported against leaf-spotting fungal and bacterial pathogens, systemic viruses, a crown-rotting fungal pathogen, root-knot nematodes, and a stem-blight fungal pathogen as well as damping-off, blue mold, and late blight diseases.
“Induced Systemic Resistance and Promotion of Plant Growth by Bacillus spp.” In Phytopathology. 2004. 94(11): 1259-1266.
Bacillus cereus
diverse B. cereus strains isolated from soil samples from five continents produce antibiotic B and that antibiotic B contributes to suppression of plant disease.
“Production of kanosamine by Bacillus cereus UW85.” in Applied Environmental Microbiology. 1996. 62 (8): 3061–3065.
Bacillus cereus
Bacillus cereus interacts with numerous other microorganisms in the rhizosphere. B. cereus strains suppress plant diseases caused by oomycete (protist) pathogens and enhance crop productivity
“Peptidoglycan from Bacillus cereus Mediates Commensalism with Rhizosphere Bacteria from the Cytophaga-Flavobacterium Group.” In Applied Environmental Microbiology. 2006. 72(8): 5421–5427.
Bacillus cereus
Zwittermicin A is…first identified for its role in suppression of plant disease by B. cereus UW85. It has a broad spectrum of activity, inhibiting certain gram-positive, gram-negative, and eukaryotic microorganisms. It also potentiates the insecticidal activity of the protein toxin produced by Bacillus thuringiensis, increasing mortality of insects that are typically recalcitrant to killing, such as gypsy oths reared on willow leaves.
“Genetics of Zwittermicin A Production by Bacillus cereus.” in Applied Environmental Microbiology. 2004. 70(1): 104–113
Voodoo Juice promotes growth of beneficial bacteria and fungi
Bacillus amyloliquefaciens
In addition to the catabolism of inositol, a product of phytate degradation, these bacteria are actively involved in the colonization of symbiotic Rhizobium strains and Sinorhizobium fredii.
“Production and characterization of thermostable alkaline phytase from Bacillus laevolacticus isolated from rhizosphere soil.” in Journal of Industrial Microbiology & Biotechnology. 34(1): 1367-5435
Bacillus cereus
... B. cereus directly stimulates the growth of [Cytophaga-Flavobacterium] rhizosphere isolates in root exudate medium. Several lines of evidence indicate that peptidoglycan produced by B. cereus stimulates growth of CF species by serving as a source of carbon and energy.
“Peptidoglycan from Bacillus cereus Mediates Commensalism with Rhizosphere Bacteria from the Cytophaga-Flavobacterium Group.” In Applied Environmental Microbiology. 2006. 72(8): 5421–5427.
Bacillus spp.
“Garbaye and Bowen (1989) postulated that true ‘helper bacteria’ were (i) adapted to living in close association with the mycorrhizal fungus (e.g. bacteria embedded in the outer mantle in ectomycorrhizal fungi), and (ii) that their effects on mycorrhizal fungi were fungus-specific. Tremendous success in enhancing plant growth has been obtained when helper bacteria were used in conjunction with ectomycorrhizal fungi.”
“Bacteria associated with Glomus clarum spores influence mycorrhizal activity” in Soil Biology and Biochemistry. 2003. 35(3): 471-478
Bacillus spp.
These biopolymer-degrading bacteria are possibly feeding on the outer hyaline spore layer that is present in both species [of fungi] and consists mainly of chitin.
“Bacteria Associated with Spores of the Arbuscular Mycorrhizal Fungi Glomus geosporum and Glomus constrictum” in Applied and Environmental Microbiology. 2005. 71(11): 6673-6679
Voodoo Juice helps plants have extra phosphorous intake
Bacillus amyloliquefaciens
The soil plant rhizosphere bacteria, also termed as plant-growth-promoting-rhizobacteria (PGPR), are known to mobilize soil inorganic polyphosphates and/or phytate. Some of the PGPR strains of Bacillus amyloliquefaciens, isolated from soil rhizosphere, are known to be active phytases producers.
“Production and characterization of thermostable alkaline phytase from Bacillus laevolacticus isolated from rhizosphere soil.” in Journal of Industrial Microbiology & Biotechnology. 34(1): 1367-5435
Bacillus amyloliquefaciens
Application of [B. amyloliquefaciens] strain FZB45 under conditions of limited availability of phosphate might be favourable because of its high phytase productivity and the high stability of the secreted enzyme. Moreover, we have obtained first experimental hints that FZB45 is able to colonize plant roots and to express a high local concentration of phytase at the surface of the roots (E. E. Idriss, unpublished observation), making this bacterium very suitable for application as a PGPR.
“Extracellular phytase activity of Bacillus amyloliquefaciens FZB45 contributes to its plant-growth-promoting effect.” In Microbiology. 2002. 148: 2097-2109.
Bacillus amyloliquefaciens
These genetic and biochemical experiments provide strong evidence that phytase activity of B. amyloliquefaciens FZB45 is important for plant growth stimulation under phosphate limitation.
“Extracellular phytase activity of Bacillus amyloliquefaciens FZB45 contributes to its plant-growth-promoting effect.” In Microbiology. 2002. 148: 2097-2109.
Bacillus laevolacticus
Bacillus laevolacticus is a good source of thermostable alkaline phytases insensitive to the presence of inorganic phosphate in the medium that produces high levels of phytase as compared to other reported wild type strains of Bacillus sp.
“Production and characterization of thermostable alkaline phytase from Bacillus laevolacticus isolated from rhizosphere soil.” in Journal of Industrial Microbiology & Biotechnology. 34(1): 1367-5435
Bacillus laevolacticus
Phytase [myo-inositol hexakis(dihydrogenphosphate) phosphohydrolase, EC 3.1.3.8] catalyses the hydrolysis of phytic acid to myo-inositol and phosphoric acid via penta- to mono-phosphates. This enzymatic activity produces available phosphate and a non-metal chelator compound.
“Phytase activity in sourdough lactic acid bacteria: purification and characterization of a phytase from Lactobacillus sanfranciscensis CB1.” In International Journal of Food Microbiology. 2003. 87(3): 259-270.
Bacillus laevolacticus
Phytase activities of bacteria inhabiting the plant rhizosphere may contribute to their plant growth-promoting effect (23, 44).
“Dual Role of the PhoP~P Response Regulator: Bacillus amyloliquefaciens FZB45 Phytase Gene Transcription Is Directed by Positive and Negative Interactions with the phyC Promoter.” In Journal of Bacteriology. 2006. 188(19): 6953-6965
Removes nutrient chelating agent
Bacillus laevolacticus
Phytase [myo-inositol hexakis(dihydrogenphosphate) phosphohydrolase, EC 3.1.3.8] catalyses the hydrolysis of phytic acid to myo-inositol and phosphoric acid via penta- to mono-phosphates. This enzymatic activity produces available phosphate and a non-metal chelator compound.
“Phytase activity in sourdough lactic acid bacteria: purification and characterization of a phytase from Lactobacillus sanfranciscensis CB1.” In International Journal of Food Microbiology. 2003. 87(3): 259-270.
Bacillus laevolacticus
Besides their ability to make phytate phosphorus available, elimination of chelate-forming phytate, which is known to bind nutritionally important minerals (Zn2+, Fe2+, and Ca2+), is another beneficial effect of extracellular phytase activities of several soil bacteria, such as Pseudomonas (24), Klebsiella (18), and Bacillus spp. (25).
“Dual Role of the PhoP~P Response Regulator: Bacillus amyloliquefaciens FZB45 Phytase Gene Transcription Is Directed by Positive and Negative Interactions with the phyC Promoter.” In Journal of Bacteriology. 2006. 188(19): 6953-6965
- when empty chelators are naturally present in soil, they will bind nutrients and prevent them from being absorbed by your plants. Phytase breaks apart soil-locked nutrients.
Fixes nitrogen
Paenibacillus azotofixans/B>
[Paenibacillus azotofixans strains] all effectively fix nitrogen in vitro even in the presence of nitrate (Seldin et al. 1984), some strains produce antimicrobial substances (Seldin & Penido 1990) and solubilize organic phosphates.
“Evaluation of Biolog system for identification of strains of Paenibacillus Azotofixans.” In Antonie van Leeuwenhoek. 1997. 71(3): 195–200
Paenibacillus azotofixans
Strains belonging to the species Paenibacillus azotofixans were shown to be efficient nitrogen fixers prevalent in the rhizospheres of maize, sorghum, sugarcane, wheat, banana, and forage grasses. Some strains are able to produce antimicrobial substances and solubilize organic phosphates.
“Comparison of Paenibacillus azotofixans Strains Isolated from Rhizoplane, Rhizosphere, and Non-Root-Associated Soil from Maize Planted in Two Different Brazilian Soils.” In Applied Environmental Microbiology. 1998. 64(10): 3860–3868.
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