Stable organic-carrier-based microbial inoculants and method for producing the same

Abstract

The present invention relates to methods for producing microbial preparations for crop production and microbial preparations produced through such methods. The formulations contain microbial inoculants like Azospirillum spp. which are grown in a complex medium, lyophilized with powdered milk and further supplemented with minerals and vitamins, starch and inert substances such as talc, chalk and diatomaceous earth. The products manufactured by such methods have shelf-lives of more than 3 years. They are used to inoculate seeds, seedlings and planting materials among different agriculture and forestry species for enhancing root, root hair an shoot formation, enabling plants to efficiently use soil nutrients, supplying plants with nitrogen from the air, tolerating drought and protecting plants from root pathogenes.

1 . A method for producing and formulating microbial inoculants with a prolonged shelf-life, comprising:
 isolating a microbial colony from a culture of a bacterial microorganism;   growing the colony in an organic medium;   harvesting the microbial cells from the organic medium;   mixing the harvested cells with milk and glycerol;   drying the microbial cells by lyophilization;   grinding the dried microbial cells into powder; and   mixing the microbial powder with nutrients, anti-contaminant and a dessicant.   
   
   
       2 . The method according to  claim 1  wherein the organic medium consists of a mixture of high protein and vitamin-supplemented feed formula. 
   
   
       3 . The method according to  claim 1  wherein harvesting of the microbial cells comprises filtration or centrifugation. 
   
   
       4 . The method according to  claim 1  wherein the anti-contaminant agent is an anti-fungal agent. 
   
   
       5 . The method according to  claim 1  wherein the pest protectant agent is an anti-viral agent. 
   
   
       6 . The method according to  claim 1  wherein the nutrients mixed with the microbial powder is selected from the group consisting of vitamins, minerals and flour and, wherein the dessicant is selected from the group consisting of talc, chalk and diatomaceous earth. 
   
   
       7 . (canceled) 
   
   
       8 . The method according to  claim 1  wherein the bacterial microorganism is  Azospirillum halopraeferens.    
   
   
       9 . The method according to  claim 1  wherein the bacterial microorganism is  Azospirillum lipoferum.    
   
   
       10 . The method according to  claim 1  wherein the bacterial microorganism is  Bacillus pumilus.    
   
   
       11 . The method according to  claim 1  wherein the bacterial microorganism is  Azospirillum  SC/PH. 
   
   
       12 . The method according to  claim 1  wherein the bacterial microorganism is  Azospirillum  AH/EH. 
   
   
       13 . A biologically pure culture of  Azospirillum  SC/PH produced by the method comprising:
 isolating a microbial colony from a culture of a bacterial microorganism, wherein the microorganism is  Azospirillum  SC/PH;   growing the colony in an organic medium;   harvesting the microbial cells from the organic medium;   mixing the harvested cells with milk and glycerol;   drying the microbial cells by lyophilization;   grinding the dried microbial cells into powder; and   mixing the microbial powder with nutrients, anti-contaminant and a dessicant.   
   
   
       14 . A biologically pure culture of  Azospirillum  AH/EH produced by the method comprising:
 isolating a microbial colony from a culture of a bacterial microorganism, wherein the microorganism is  Azospirillum  AH/EH;   growing the colony in an organic medium;   harvesting the microbial cells from the organic medium;   mixing the harvested cells with milk and glycerol;   drying the microbial cells by lyophilization;   grinding the dried microbial cells into powder; and   mixing the microbial powder with nutrients, anti-contaminant and a dessicant.   
   
   
       15 . The biologically pure culture of  claim 13  comprising a shelf life of at least 36 months or 3 years. 
   
   
       16 . The biologically pure culture of  claim 14  comprising a shelf life of at least 36 months or 3 years. 
   
   
       17 . The biologically pure culture of  Azospirillum  SC/PH of  claim 13 , wherein the organic medium comprises a mixture of high protein and vitamin-supplemented feed formula. 
   
   
       18 . The biologically pure culture of  Azospirillum  SC/PH of  claim 13 , wherein harvesting of the microbial cells comprises filtration or centrifugation. 
   
   
       19 . The biologically pure culture of  Azospirillum  SC/PH of  claim 13 , wherein the anti-contaminant agent is an anti-fungal agent. 
   
   
       20 . The biologically pure culture of  Azospirillum  SC/PH of  claim 13 , wherein the pest protectant agent is an anti-viral agent. 
   
   
       21 . The biologically pure culture of  Azospirillum  SC/PH of  claim 13 , wherein the nutrients mixed with the microbial powder is selected from the group consisting of vitamins, minerals and flour and wherein, the dessicant is selected from group consisting of talc, chalk and diatomaceous earth. 
   
   
       22 . The biologically pure culture of  Azospirillum  AH/EH of  claim 14 , wherein the organic medium comprises a mixture of high protein and vitamin-supplemented feed formula. 
   
   
       23 . The biologically pure culture of  Azospirillum  AH/EH of  claim 14 , wherein harvesting of the microbial cells comprises filtration or centrifugation. 
   
   
       24 . The biologically pure culture of  Azospirillum  AH/EH of  claim 14 , wherein the anti-contaminant agent is an anti-fungal agent. 
   
   
       25 . The biologically pure culture of  Azospirillum  AH/EH of  claim 14 , wherein the pest protectant agent is an anti-viral agent. 
   
   
       26 . The biologically pure culture of  Azospirillum  AH/EH of  claim 14 , wherein the nutrients mixed with the microbial powder is selected from the group consisting of vitamins, minerals and flour and wherein, the dessicant is selected from group consisting of talc, chalk and diatomaceous earth. 
   
   
       27 . The bacterial microorganism of  claim 8 , wherein a representative sample of  Azospirillum halopraeferens  was deposited under UPCC 1370. 
   
   
       28 . The bacterial microorganism of  claim 9 , wherein a representative sample of  Azospirillum lipoferum  was deposited under UPCC 1371. 
   
   
       29 . The bacterial microorganism of  claim 10 , wherein a representative sample of  Bacillus pumilus  was deposited under UPCC 1372. 
   
   
       30 . The bacterial microorganism of  claim 11 , wherein a representative sample of  Azospirillum  SC/PH was deposited under NRRL B-50317. 
   
   
       31 . The bacterial microorganism of  claim 12 , wherein a representative sample of  Azospirillum  AH/EH was deposited under NRRL B-50318.