Method for propagating microorganisms within plant bioreactors and stably storing microorganisms within agricultural seeds

Patent metadata

Field	Value
Publication number	US-2020008431-A1
Application number	US-201916512219-A
Country	US
Kind code	A1
Filing date	Jul 15, 2019
Priority date	Dec 24, 2013
Publication date	Jan 9, 2020
Grant date	—

Abstract

The present invention relates to methods of scalably producing microorganisms by propagating them within plant tissues and introducing them into agricultural seeds to improve their shelf-life during long-term storage, to produce substances of interest, and to create libraries of microbes.

First claim

1 .- 71 . (canceled)

72 . A method of producing a substance selected from the group consisting of a Hydroquinone glucosyltransferase, a Replication factor C subunit 3, a NEDD8-activating enzyme E1 catalytic subunit, and a Delta3,5-delta2, 4-dienoyl-CoA isomerase within a bacterial colonized cereal plant bioreactor, the method comprising:
a. germinating a cereal seed into which at least one inoculant bacterial endophyte has been introduced, wherein the bacterial endophyte is from the genus Burholderia, comprises a 16S rRNA nucleic acid sequence comprising SEQ ID NO: 1446 and can grow on nitrogen free media, to produce the bacterial colonized cereal plant bioreactor; and b. growing the bacterial colonized cereal plant bioreactor under conditions such that the bacterial endophyte proliferates and the substance is produced in the bacterial colonized cereal plant bioreactor, wherein the substance is produced at a higher concentration in the bacterial colonized cereal plant bioreactor compared to an isoline plant grown from a non-inoculated cereal seed.

73 . The method of claim 72 , wherein the at least one inoculant bacterial endophyte is introduced into the cereal seed by the method of: contacting at least one flower of a cereal plant in the course of a flowering phase of the cereal plant with a preparation comprising a population of the at least one inoculant bacterial endophyte, wherein the preparation comprises 10 6 to 10 9 CFU per ml, whereby the inoculant bacterial endophytes enter the flowering plant via at least one flower and are conveyed to an interior of at least one seed produced by the flowering plant.

74 . The method of claim 72 , wherein the cereal plant is corn.

75 . The method of claim 72 , wherein the cereal plant is wheat.

76 . The method of claim 72 , wherein the substance improves symbiosis.

77 . The method of claim 72 , wherein the substance is the Hydroquinone glucosyltransferase.

78 . The method of claim 72 , wherein the substance is the Replication factor C subunit 3.

79 . The method of claim 72 , wherein the substance is the NEDD8-activating enzyme E1 catalytic subunit.

80 . The method of claim 72 , wherein the substance is the Delta3,5-delta2, 4-dienoyl-CoA isomerase.

81 . The method of claim 73 , wherein the substance is the Hydroquinone glucosyltransferase.

82 . The method of claim 73 , wherein the substance is the Replication factor C subunit 3.

83 . The method of claim 73 , wherein the substance is the NEDD8-activating enzyme E1 catalytic subunit.

84 . The method of claim 73 , wherein the substance is the Delta3,5-delta2, 4-dienoyl-CoA isomerase.

Assignees

• Indigo Ag Inc
• Ait Austrian Inst Tech Gmbh

Inventors

• Mitter Birgit
• Naveed Muhammad
• Berninger Teresa
• Compant Stephane
• Sessitsch Angela
• Von Maltzahn Geoffrey
• Flavell Richard Bailey
• Toledo Gerardo V
• Djonovic Slavica
• Marquez Luis Miguel
• Johnston David Morris
• Millet Yves Alain
• Lyford Jeffrey
• Leff Jonathan W
• Samayoa Phillip
• Sadowski Craig

Classifications

• G01N33/0098

  Plants or trees (wood G01N33/46) · CPC title

• A01N63/30

  Microbial fungi; Substances produced thereby or obtained therefrom · CPC title

• A01H5/00

  Angiosperms, i.e. flowering plants, characterised by their plant parts; Angiosperms characterised otherwise than by their botanic taxonomy · CPC title

• A01N63/00 Primary

  Biocides, pest repellants or attractants, or plant growth regulators containing microorganisms, viruses, microbial fungi, animals or substances produced by, or obtained from, microorganisms, viruses, microbial fungi or animals, e.g. enzymes or fermentates (containing compounds of determined constitution A01N27/00 - A01N59/00; unicellular algae A01N65/03) · CPC title

• C12N1/20 Primary

  Bacteria; Culture media therefor · CPC title