System for delivery of microbial inoculants and related materials and methods

Patent metadata

Field	Value
Publication number	US-10774298-B2
Application number	US-201414781545-A
Country	US
Kind code	B2
Filing date	Dec 10, 2014
Priority date	Dec 11, 2013
Publication date	Sep 15, 2020
Grant date	Sep 15, 2020

Abstract

Described herein are systems, methods and kits for culturing and applying fresh microbial inoculants in the field. The system, methods and kits are easy to use, reliable, sealed to prevent contamination, and can be stored on location, for use on demand by those unskilled in the microbiological arts. Particular embodiments described herein may be used to increase agricultural crop yields.

First claim

What is claimed is:

1. A microbial inoculant system, comprising:
a first sealed chamber including;
aerobic microbes stabilized on a solid matrix wherein the solid matrix comprises ground soybean seeds;
a disruptable dividing member defining a wall of the first chamber and configured to release the contents of the first chamber when disrupted; and
a second sealed chamber adjacent to and sealed by connection to the first sealed chamber, wherein the second sealed chamber is configured to receive the contents of the first sealed chamber when the dividing member is disrupted and includes nutrient media suitable for growing the microbes through a log phase growth stage; wherein the second sealed chamber comprises a gas permeable member to provide oxygen levels to support log phase growth of the aerobic microbes; wherein the first sealed chamber is removably connected to the second sealed chamber, wherein one or both of the first and second sealed chambers are adapted for threading or snapping the chambers together;
and wherein the system is configured to be activated by disrupting the dividing member using one of the following disrupting means:
(1) a piercing device affixed to an inner surface of a wall of the first sealed chamber in a position opposing the dividing member, wherein the wall is configured to move the piercing device towards the dividing member when a force is applied to an external surface of the wall, wherein the force consists of pushing the external surface of the wall inwardly at the fixation site of the piercing device;
(2) a piercing device in the form of a ball bearing or bead present in one or both of the first and second sealed chambers; and
(3) wherein the dividing member is made of a partially dissolvable material capable of breaking when the system is vigorously shaken, but which remains intact when exposed to a liquid media during normal handling.

2. The system of claim 1 , wherein the aerobic microbes, when stabilized on the matrix, are in a form selected from one or more of: planktonic, biofilmic, dormant, lyophilized, partially dormant, partially lyophilized, sporulated, dehydrated, and freeze-dried; and, wherein the microbes are not present in a log phase growth.

3. The system of claim 1 , wherein the media present in the second chamber comprises water or other liquid media that support the growth of the aerobic microbes present in the first chamber.

4. The system of claim 1 , wherein the percent of media in the second chamber, as a function of total volume of the second chamber, is from about 35% to about 65% to allow for gas exchange to support proper growth of the aerobic microbes.

5. The system of claim 1 , further having at least one access port for connection to a delivery device located in the second chamber distal to the dividing membrane.

6. The system of claim 1 , wherein the aerobic microbes are selected from the group consisting of one or more aerobic bacteria.

7. The system of claim 6 , wherein the one or more aerobic bacteria are selected from the group consisting of Achromobacter, Arthrobacter, Azotobacter, Azospirillum, Bacillus, Bradyrhizobium, Brevibacillus, Burkholderia, Chitinophaga, Chryseobacterium, Curtobacterium, Klebsiella, Methylobacterium, Mitsuaria, Novosphingomonas, Paenibacillus, Pantoea, Pasteuria, Pseudomonas, Rhizobium, Serratia, Sinorhizobium, Sphingomonas, Stenotrophomonas , and Streptomyces.

8. The system of claim 1 , wherein the second chamber of the system is configured to remain sealed after disruption of the dividing member to resist passive environmental contamination during the log phase growth of the aerobic microbes in the second chamber.

9. The system of claim 1 , wherein the second chamber comprises a neck and the first chamber comprises a substantially cylindrical side wall configured for removable connection to an outer surface of the neck.

10. The system of claim 1 , wherein the second sealed chamber comprises side walls configured to allow the system to be stored on its side.

11. The system of claim 1 , wherein the gas permeable member is a material selected from liquid-permeable or impermeable, hydrophobic or hydrophilic, porous or non-porous materials.

12. The system of claim 7 , wherein the one or more aerobic bacteria are selected from the group consisting of Azospirillum, Mitsuaria , and Pseudomonas.

13. A method of promoting plant growth by treating seeds, seedlings, plants, or soil, comprising:
providing a microbial inoculant system according to claim 1
disrupting the integrity of the dividing member to introduce the microbes from the first sealed chamber into the nutrient media in the second sealed chamber,
culturing the aerobic microbes introduced into the second chamber under conditions suitable for rapid for rapid growth of the microbes; and
applying the cultured aerobic microbes to seeds, seedlings, plants, soil, or a combination thereof.

14. The method of claim 13 , wherein the cultured aerobic microbes are diluted to a cell number of about 1,000 to about 10,000,000 CFUs per ml in the target diluents prior to applying to the seeds, seedlings, plants, soil or a combination thereof.

15. The method of claim 13 , wherein applying cultured aerobic microbes is via spraying at a rate of about 103 cells/ml to about 107 cells/ml.

16. The method of claim 15 , wherein applying cultured aerobic microbes is via spraying at a rate of about 105 cells per ml.

17. The method of claim 13 , wherein applying cultured aerobic microbes results in about 1000 cells per seed, seedling, plant, or square inch of soil, to about 10 million cells per seed, seedling, plant, or square inch of soil.

Assignees

• 3Bar Biologics Inc
• Ohio State Innovation Foundation

Inventors

• Caldwell Bruce H
• Mcspadden Gardener Brian B

Classifications

• C12M37/04 Primary

  Seals · CPC title

• A01N63/27 Primary

  Pseudomonas · CPC title

• A01N63/22

  Bacillus · CPC title

• A01N63/20

  Bacteria; Substances produced thereby or obtained therefrom · CPC title

• A01C1/06

  Coating or dressing seed · CPC title