IP Library › Granted Patent US 12,692,203
Granted Patent B2
US 12,692,203 · App. 19/242,173 · Granted Jul 28, 2026

Temporally and spatially targeted dynamic nitrogen delivery by remodeled microbes

Inventors: Alvin Tamsir (Berkeley, CA); Sarah Bloch (Berkeley, CA); Mark Reisinger (Berkeley, CA); Ernest Sanders (Berkeley, CA); Richard Broglie (Berkeley, CA); Rosemary Clark (Berkeley, CA); Karsten Temme (Berkeley, CA)
Assignee: Pivot Bio, Inc.
C05C3/00A01H1/12A01H6/4684C05C9/005C05F11/08
View Patent ↗
Loading inventors, assignments & file history…
Monitor This Case
Get email alerts when status or documents change.
Order Certified Copies
Most orders are placed with the USPTO same day — all within 24 business hours.
Order via The Patent Place →
Pre-filled with this patent's details
Quick Facts
Patent No.
US 12,692,203
App. No.
19/242,173
Filed
Jun 18, 2025
Granted
Jul 28, 2026
Kind
B2
Art Unit
1655
USPC
435/410
Abstract

The present disclosure provides non-intergeneric remodeled microbes that are able to fix atmospheric nitrogen and deliver such to plants in a targeted, efficient, and environmentally sustainable manner. The utilization of the taught microbial products will enable farmers to realize more productive and predictable crop yields without the nutrient degradation, leaching, or toxic runoff associated with traditional synthetically derived nitrogen fertilizer, by mitigating or eliminating the need for exogenous nitrogen-containing fertilizers. The remodeled microbes have unique colonization and nitrogen fixation abilities, which enable the microbes to deliver nitrogen to a cereal plant in a spatially targeted (e.g. rhizospheric) and temporally targeted (e.g. during advantageous stages of plant's life cycle) manner. The microbes are able to replace the standard agricultural practice of sidedressing and enable a more environmentally sustainable form of farming. The present disclosure also provides methods of using non-intergeneric remodeled microbes, for example, to fix atmospheric nitrogen by reducing or eliminating the need for exogenous nitrogen-containing fertilizers, to increase yield, and to reduce infield variability in the yield.

Claims (24)

1 . An agricultural composition engineered for reduced variability in mean corn yield, comprising:

a) a plurality of corn seeds; and

b) a plurality of non-intergeneric remodeled bacteria coated onto said plurality of corn seeds, wherein members of the plurality of non-intergeneric remodeled bacteria produce fixed N of at least about 5.49×10 −13 mmol of N per CFU per hour, and

wherein members of the plurality of non-intergeneric remodeled bacteria are capable of colonizing corn plants upon planting of said corn seeds and supplying the corn plants with fixed N, and

wherein the standard deviation of mean corn yield measured across a locus upon planting of said corn seeds is lower, as compared to a control plurality of corn seeds planted without said plurality of non-intergeneric remodeled bacteria.

2 . The agricultural composition of claim 1 , wherein members of the plurality of non-intergeneric remodeled bacteria produce in the aggregate at least about 15 pounds of fixed N per acre over the course of at least about 10 days to about 60 days upon planting of said corn seeds.

3 . The agricultural composition of claim 1 , wherein members of the plurality of non-intergeneric remodeled bacteria each produce fixed N of at least about 2.75×10 −12 mmol of N per CFU per hour.

4 . The agricultural composition of claim 1 , wherein members of the plurality of non-intergeneric remodeled bacteria are capable of colonizing the root surface of corn plants upon planting of said corn seeds.

5 . The agricultural composition of claim 1 , wherein members of the plurality of non-intergeneric remodeled bacteria comprise at least one genetic variation introduced into at least one gene, or non-coding polynucleotide, involved in a pathway selected from the group consisting of: the nitrogen fixation or assimilation genetic regulatory network, exopolysaccharide production, endo-polygalaturonase production, trehalose production, and glutamine conversion.

6 . The agricultural composition of claim 1 , wherein members of the plurality of non-intergeneric remodeled bacteria comprise at least one genetic variation introduced into at least one gene, or non-coding polynucleotide, involved in the nitrogen fixation pathway.

7 . The agricultural composition of claim 1 , wherein members of the plurality of non-intergeneric remodeled bacteria comprise at least one genetic variation introduced into at least one gene, or non-coding polynucleotide, involved in the nitrogen assimilation pathway.

8 . The agricultural composition of claim 1 , wherein members of the plurality of non-intergeneric remodeled bacteria comprise at least one genetic variation introduced into at least one gene, or non-coding polynucleotide, involved in the glutamine conversion pathway.

9 . The agricultural composition of claim 1 , wherein members of the plurality of non-intergeneric remodeled bacteria comprise an introduced control sequence operably linked to at least one gene of the nitrogen fixation pathway.

10 . The agricultural composition of claim 1 , wherein members of the plurality of non-intergeneric remodeled bacteria comprise an introduced control sequence operably linked to at least one gene of the nitrogen assimilation pathway.

11 . The agricultural composition of claim 1 , wherein members of the plurality of non-intergeneric remodeled bacteria comprise a heterologous promoter operably linked to at least one gene of the nitrogen fixation pathway.

12 . The agricultural composition of claim 1 , wherein members of the plurality of non-intergeneric remodeled bacteria comprise a heterologous promoter operably linked to at least one gene of the nitrogen assimilation pathway.

13 . The agricultural composition of claim 1 , wherein members of the plurality of non-intergeneric remodeled bacteria comprise at least one genetic variation introduced into: nifA, nifL, ntrB, ntrC, polynucleotide encoding glutamine synthetase, glnA, glnB, glnK, drat, amtB, polynucleotide encoding glutaminase, glnD, glnE, nifJ, nifH, nifD, nifK, nifY, nifE, nifN, nifU, nifS, nifV, nifW, nifZ, nifM, nifF, nifB, nifQ, a gene associated with biosynthesis of a nitrogenase enzyme, bcsii, bcsiii, yjbE, fhaB, pehA, otsB, treZ, glsA2, or combinations thereof.

14 . The agricultural composition of claim 1 , wherein members of the plurality of non-intergeneric remodeled bacteria comprise at least one genetic variation introduced into: nifA, nifL, rpoN, glnE, glnA, glsA2, amtB, bcsii, bcsiii, pehA, fhaB, dctA, phoB1, glnD, glgA, or combinations thereof.

15 . The agricultural composition of claim 1 , wherein members of the plurality of non-intergeneric remodeled bacteria comprise at least one genetic variation introduced into glnA.

16 . The agricultural composition of claim 1 , wherein members of the plurality of non-intergeneric remodeled bacteria comprise at least one genetic variation introduced into at least one gene, or non-coding polynucleotide, of the nitrogen fixation or assimilation genetic regulatory network that results in one or more of: increased expression or activity of NifA or glutaminase; decreased expression or activity of NifL, NtrB, glutamine synthetase, GlnB, GlnK, DraT, AmtB; decreased adenylyl-removing activity of GlnE; or decreased uridylyl-removing activity of GlnD.

17 . The agricultural composition of claim 1 , wherein members of the plurality of non-intergeneric remodeled bacteria comprise at least one of: a mutated nifL gene that has been altered to comprise a heterologous promoter inserted into said nifL gene; a mutated glnE gene that results in a truncated GlnE protein lacking an adenylyl-removing (AR) domain; a mutated amtB gene that results in the lack of expression of said amtB gene; or combinations thereof.

18 . The agricultural composition of claim 1 , wherein members of the plurality of non-intergeneric remodeled bacteria comprise bacteria selected from: Rahnella aquatilis, Klebsiella variicola, Achromobacter spiritinus, Achromobacter marplatensis, Microbacterium murale, Kluyvera intermedia, Kosakonia pseudosacchari, Enterobacter sp., Azospirillum lipoferum, Kosakonia sacchari , or combinations thereof.

19 . The agricultural composition of claim 1 , wherein members of the plurality of non-intergeneric remodeled bacteria comprise bacteria selected from: bacteria deposited as NCMA 201701002, bacteria deposited as NCMA 201708004, bacteria deposited as NCMA 201708003, bacteria deposited as NCMA 201708002, bacteria deposited as NCMA 201712001, bacteria deposited as NCMA 201712002, or combinations thereof.

20 . The agricultural composition of claim 1 , wherein members of the plurality of non-intergeneric remodeled bacteria comprise bacteria with a nucleic acid sequence that shares at least about 90% sequence identity to a nucleic acid sequence selected from SEQ ID NOs: 177-260, 296-303, and 458-469.

Assignments (1)
ASSIGNMENT OF ASSIGNOR'S INTEREST Recorded Jun 24, 2025
From: TAMSIR, ALVIN; BLOCH, SARAH; REISINGER, MARK; SANDERS, ERNEST; BROGLIE, RICHARD; CLARK, ROSEMARY; TEMME, KARSTEN
To: PIVOT BIO, INC.
Reel/Frame 071707/0855 →
Continuity (4)
Continuation 17258024 · Jul 11, 2019
Provisional Application 62801504 · Feb 5, 2019
Provisional Application 62696452 · Jul 11, 2018
Related Publication 20250376424A1 · Dec 11, 2025
References Cited (400)
US 1520545A · Murphy · 1924 [cited by applicant]
US 4782022A · Puhler et al. · 1988 [cited by applicant]
US 4832728A · Allan et al. · 1989 [cited by applicant]
US 4970147A · Huala et al. · 1990 [cited by applicant]
US 5071743A · Slilaty et al. · 1991 [cited by applicant]
US 5188960A · Payne et al. · 1993 [cited by applicant]
US 5229291A · Nielsen et al. · 1993 [cited by applicant]
US 5354670A · Nickoloff et al. · 1994 [cited by applicant]
US 5427785A · Ronson et al. · 1995 [cited by applicant]
US 5780270A · Lesley · 1998 [cited by applicant]
US 5789166A · Bauer et al. · 1998 [cited by applicant]
US 5877012A · Estruch et al. · 1999 [cited by applicant]
US 5880275A · Fischhoff et al. · 1999 [cited by applicant]
US 5916029A · Smith et al. · 1999 [cited by applicant]
US 6033861A · Schafer et al. · 2000 [cited by applicant]
US 6033874A · Baum et al. · 2000 [cited by applicant]
US 6083499A · Narva et al. · 2000 [cited by applicant]
US 6107279A · Estruch et al. · 2000 [cited by applicant]
US 6114148A · Seed et al. · 2000 [cited by applicant]
US 6127180A · Narva et al. · 2000 [cited by applicant]
US 6137033A · Estruch et al. · 2000 [cited by applicant]
US 6218188B1 · Cardineau et al. · 2001 [cited by applicant]
US 6248535B1 · Danenberg et al. · 2001 [cited by applicant]
US 6326351B1 · Donovan et al. · 2001 [cited by applicant]
US 6340593B1 · Cardineau et al. · 2002 [cited by applicant]
US 6391548B1 · Bauer et al. · 2002 [cited by applicant]
US 6399330B1 · Donovan et al. · 2002 [cited by applicant]
US 6548289B1 · Beynon et al. · 2003 [cited by applicant]
US 6548291B1 · Narva et al. · 2003 [cited by applicant]
US 6596509B1 · Bauer et al. · 2003 [cited by applicant]
US 6624145B1 · Narva et al. · 2003 [cited by applicant]
US 6673610B2 · Miyawaki et al. · 2004 [cited by applicant]
US 6713063B1 · Malvar et al. · 2004 [cited by applicant]
US 6713285B2 · Bauer et al. · 2004 [cited by applicant]
US 6773900B2 · Short et al. · 2004 [cited by applicant]
US 6841358B1 · Locht et al. · 2005 [cited by applicant]
US 6949626B2 · Donovan et al. · 2005 [cited by applicant]
US 6962705B2 · Malvar et al. · 2005 [cited by applicant]
US 7064249B2 · Corbin et al. · 2006 [cited by applicant]
US 7070982B2 · Malvar et al. · 2006 [cited by applicant]
US 7084331B2 · Isawa et al. · 2006 [cited by applicant]
US 7105332B2 · Abad et al. · 2006 [cited by applicant]
US 7132265B2 · Bauer et al. · 2006 [cited by applicant]
US 7244820B2 · Miles et al. · 2007 [cited by applicant]
US 7329736B2 · Abad et al. · 2008 [cited by applicant]
US 7374753B1 · Farmer et al. · 2008 [cited by applicant]
US 7378499B2 · Abad et al. · 2008 [cited by applicant]
US 7385107B2 · Donovan et al. · 2008 [cited by applicant]
US 7419698B2 · Jones · 2008 [cited by applicant]
US 7438901B2 · Loessner et al. · 2008 [cited by applicant]
US 7449552B2 · Abad et al. · 2008 [cited by applicant]
US 7462760B2 · Abad et al. · 2008 [cited by applicant]
US 7470427B2 · Cocking · 2008 [cited by applicant]
US 7476781B2 · Abad et al. · 2009 [cited by applicant]
US 7485451B2 · Vandergheynst et al. · 2009 [cited by applicant]
US 7491698B2 · Hey et al. · 2009 [cited by applicant]
US 7491869B2 · Abad et al. · 2009 [cited by applicant]
US 7504229B2 · Donovan et al. · 2009 [cited by applicant]
US 7615686B2 · Miles et al. · 2009 [cited by applicant]
US 7803943B2 · Mao et al. · 2010 [cited by applicant]
US 7858849B2 · Cerf et al. · 2010 [cited by applicant]
US 7923602B2 · Carozzi et al. · 2011 [cited by applicant]
US 8076142B2 · Huang et al. · 2011 [cited by applicant]
US 8084416B2 · Sampson et al. · 2011 [cited by applicant]
US 8084418B2 · Hey et al. · 2011 [cited by applicant]
US 8137665B2 · Cocking · 2012 [cited by applicant]
US 8236757B2 · Carozzi et al. · 2012 [cited by applicant]
US 8237020B2 · Miles et al. · 2012 [cited by applicant]
US 8268584B1 · Harwood et al. · 2012 [cited by applicant]
US 8304604B2 · Lira et al. · 2012 [cited by applicant]
US 8304605B2 · Lira et al. · 2012 [cited by applicant]
US 8318900B2 · Sampson et al. · 2012 [cited by applicant]
US 8319019B2 · Abad et al. · 2012 [cited by applicant]
US 8334366B1 · Hughes et al. · 2012 [cited by applicant]
US 8334431B2 · Sampson et al. · 2012 [cited by applicant]
US 8377671B2 · Cournac et al. · 2013 [cited by applicant]
US 8481026B1 · Woodruff et al. · 2013 [cited by applicant]
US 8513494B2 · Wu et al. · 2013 [cited by applicant]
US 8530411B2 · Cerf et al. · 2013 [cited by applicant]
US 8575433B2 · Cerf et al. · 2013 [cited by applicant]
US 8686233B2 · Cerf et al. · 2014 [cited by applicant]
US 8759619B2 · Sampson et al. · 2014 [cited by applicant]
US 8795965B2 · Zhang · 2014 [cited by applicant]
US 8802933B2 · Abad et al. · 2014 [cited by applicant]
US 8802934B2 · Abad et al. · 2014 [cited by applicant]
US 9150851B2 · Wigley et al. · 2015 [cited by applicant]
US 9321697B2 · Das et al. · 2016 [cited by applicant]
US 9487451B2 · Doty et al. · 2016 [cited by applicant]
US 9512431B2 · Mirsky et al. · 2016 [cited by applicant]
US 9657298B2 · Soto, Sr. et al. · 2017 [cited by applicant]
US 9957509B2 · Mirsky et al. · 2018 [cited by applicant]
US 9975817B2 · Temme et al. · 2018 [cited by applicant]
US 10384983B2 · Temme et al. · 2019 [cited by applicant]
US 10556839B2 · Temme et al. · 2020 [cited by applicant]
US 10662432B2 · Mirsky et al. · 2020 [cited by applicant]
US 10919814B2 · Temme et al. · 2021 [cited by applicant]
US 10934226B2 · Temme et al. · 2021 [cited by applicant]
US 10968446B2 · Zhao et al. · 2021 [cited by applicant]
US 11479516B2 · Voigt et al. · 2022 [cited by applicant]
US 11565979B2 · Temme et al. · 2023 [cited by applicant]
US 11678667B2 · Reisinger et al. · 2023 [cited by applicant]
US 11678668B2 · Reisinger et al. · 2023 [cited by applicant]
US 11739032B2 · Temme et al. · 2023 [cited by applicant]
US 11946162B2 · Zhao et al. · 2024 [cited by applicant]
US 11963530B2 · Reisinger et al. · 2024 [cited by applicant]
US 11993778B2 · Tamsir et al. · 2024 [cited by applicant]
US 12151988B2 · Tamsir et al. · 2024 [cited by applicant]
US 12209245B2 · Mirsky et al. · 2025 [cited by applicant]
US 12268212B2 · Reisinger et al. · 2025 [cited by applicant]
US 12281299B2 · Voigt et al. · 2025 [cited by applicant]
US 12281980B2 · Wood et al. · 2025 [cited by applicant]
US 12290074B2 · Reisinger et al. · 2025 [cited by applicant]
US 12391624B2 · Tamsir · 2025 [cited by examiner]
US 12421519B2 · Higgins et al. · 2025 [cited by applicant]
US 12471599B2 · Reisinger et al. · 2025 [cited by applicant]
US 12478068B2 · Rezaei et al. · 2025 [cited by applicant]
US 12497342B2 · Temme et al. · 2025 [cited by applicant]
US 20040197916A1 · Carozzi et al. · 2004 [cited by applicant]
US 20040197917A1 · Carozzi et al. · 2004 [cited by applicant]
US 20040210964A1 · Carozzi et al. · 2004 [cited by applicant]
US 20040210965A1 · Carozzi et al. · 2004 [cited by applicant]
US 20040216186A1 · Carozzi et al. · 2004 [cited by applicant]
US 20040235663A1 · Cocking · 2004 [cited by applicant]
US 20040241847A1 · Okuyama et al. · 2004 [cited by applicant]
US 20040250311A1 · Carozzi et al. · 2004 [cited by applicant]
US 20050081262A1 · Cook et al. · 2005 [cited by applicant]
US 20050266541A1 · Dillon · 2005 [cited by applicant]
US 20060112447A1 · Bogdanova et al. · 2006 [cited by applicant]
US 20060127988A1 · Wood et al. · 2006 [cited by applicant]
US 20060191034A1 · Baum et al. · 2006 [cited by applicant]
US 20060243011A1 · Someus · 2006 [cited by applicant]
US 20080295207A1 · Baum et al. · 2008 [cited by applicant]
US 20080311632A1 · Figge et al. · 2008 [cited by applicant]
US 20090105076A1 · Stewart et al. · 2009 [cited by applicant]
US 20090137390A1 · Triplett · 2009 [cited by applicant]
US 20090144852A1 · Tomso et al. · 2009 [cited by applicant]
US 20090162477A1 · Nadel et al. · 2009 [cited by applicant]
US 20090258404A1 · Mikkelsen et al. · 2009 [cited by applicant]
US 20090308121A1 · Reddy et al. · 2009 [cited by applicant]
US 20100005543A1 · Sampson et al. · 2010 [cited by applicant]
US 20100017914A1 · Hart et al. · 2010 [cited by applicant]
US 20100028870A1 · Welch et al. · 2010 [cited by applicant]
US 20100197592A1 · Heinrichs · 2010 [cited by applicant]
US 20100267147A1 · Qiao · 2010 [cited by applicant]
US 20100298211A1 · Carozzi et al. · 2010 [cited by applicant]
US 20110023184A1 · Desai et al. · 2011 [cited by applicant]
US 20110064710A1 · Benson et al. · 2011 [cited by applicant]
US 20110104690A1 · Yu et al. · 2011 [cited by applicant]
US 20110263488A1 · Carozzi et al. · 2011 [cited by applicant]
US 20120015806A1 · Paikray et al. · 2012 [cited by applicant]
US 20120107889A1 · Doty et al. · 2012 [cited by applicant]
US 20120192605A1 · McSpadden Gardener et al. · 2012 [cited by applicant]
US 20120266332A1 · Kuykendall · 2012 [cited by applicant]
US 20120278954A1 · Bowen et al. · 2012 [cited by applicant]
US 20120284813A1 · Olivier et al. · 2012 [cited by applicant]
US 20120311745A1 · Meade et al. · 2012 [cited by applicant]
US 20120311746A1 · Meade et al. · 2012 [cited by applicant]
US 20120317681A1 · Meade et al. · 2012 [cited by applicant]
US 20120317682A1 · Meade et al. · 2012 [cited by applicant]
US 20120324605A1 · Meade et al. · 2012 [cited by applicant]
US 20120324606A1 · Meade et al. · 2012 [cited by applicant]
US 20120331589A1 · Meade et al. · 2012 [cited by applicant]
US 20120331590A1 · Meade et al. · 2012 [cited by applicant]
US 20130116170A1 · Graser et al. · 2013 [cited by applicant]
US 20130126428A1 · Jones et al. · 2013 [cited by applicant]
US 20130167268A1 · Narva et al. · 2013 [cited by applicant]
US 20130167269A1 · Narva et al. · 2013 [cited by applicant]
US 20140011261A1 · Wang et al. · 2014 [cited by applicant]
US 20140155283A1 · Venkateswaran et al. · 2014 [cited by applicant]
US 20140182018A1 · Lang et al. · 2014 [cited by applicant]
US 20140223598A1 · Sampson et al. · 2014 [cited by applicant]
US 20140223599A1 · Sampson et al. · 2014 [cited by applicant]
US 20140230504A1 · Finlayson et al. · 2014 [cited by applicant]
US 20140273226A1 · Wu · 2014 [cited by applicant]
US 20140301990A1 · Gregory et al. · 2014 [cited by applicant]
US 20140329326A1 · Mirsky et al. · 2014 [cited by applicant]
US 20140336050A1 · Soto, Sr. et al. · 2014 [cited by applicant]
US 20150080261A1 · Wigley et al. · 2015 [cited by applicant]
US 20150101373A1 · Munusamy et al. · 2015 [cited by applicant]
US 20150128670A1 · Das · 2015 [cited by applicant]
US 20150237807A1 · Valiquette · 2015 [cited by applicant]
US 20150239789A1 · Kang et al. · 2015 [cited by applicant]
US 20150315570A1 · Zhao et al. · 2015 [cited by applicant]
US 20160174570A1 · Vujanovic et al. · 2016 [cited by applicant]
US 20160264929A1 · Barney et al. · 2016 [cited by applicant]
US 20160292355A1 · Lou et al. · 2016 [cited by applicant]
US 20160295868A1 · Jones et al. · 2016 [cited by applicant]
US 20170016013A1 · Fu et al. · 2017 [cited by applicant]
US 20170152519A1 · Mirsky et al. · 2017 [cited by applicant]
US 20170367349A1 · Gruver et al. · 2017 [cited by applicant]
US 20180020671A1 · Wigley et al. · 2018 [cited by applicant]
US 20180060771A1 · Mangin · 2018 [cited by applicant]
US 20180065896A1 · Van Iersel et al. · 2018 [cited by applicant]
US 20180073028A1 · Mirsky et al. · 2018 [cited by applicant]
US 20180290942A1 · Voigt et al. · 2018 [cited by applicant]
US 20190039964A1 · Temme et al. · 2019 [cited by applicant]
US 20190144352A1 · Temme et al. · 2019 [cited by applicant]
US 20190185871A1 · Zhou et al. · 2019 [cited by applicant]
US 20190325534A1 · Perry et al. · 2019 [cited by applicant]
US 20190339964A1 · Young et al. · 2019 [cited by applicant]
US 20200115715A1 · Mirsky et al. · 2020 [cited by applicant]
US 20200299637A1 · Voigt et al. · 2020 [cited by applicant]
US 20200308594A1 · Tamsir et al. · 2020 [cited by applicant]
US 20200331820A1 · Tamsir et al. · 2020 [cited by applicant]
US 20210009483A1 · Temme et al. · 2021 [cited by applicant]
US 20210163374A1 · Bloch et al. · 2021 [cited by applicant]
US 20210214282A1 · Temme et al. · 2021 [cited by applicant]
US 20210284995A1 · Zhao et al. · 2021 [cited by applicant]
US 20210315212A1 · Rezaei et al. · 2021 [cited by applicant]
US 20210345618A1 · Bloch et al. · 2021 [cited by applicant]
US 20220017911A1 · Temme et al. · 2022 [cited by applicant]
US 20220079163A1 · Reisinger et al. · 2022 [cited by applicant]
US 20220090095A1 · Higgins et al. · 2022 [cited by applicant]
US 20220106238A1 · Rezaei et al. · 2022 [cited by applicant]
US 20220127627A1 · Bloch et al. · 2022 [cited by applicant]
US 20220162544A1 · Voigt et al. · 2022 [cited by applicant]
US 20220211048A1 · Temme et al. · 2022 [cited by applicant]
US 20220282340A1 · Ryu et al. · 2022 [cited by applicant]
US 20220396530A1 · Tamsir et al. · 2022 [cited by applicant]
US 20220411344A1 · Voigt et al. · 2022 [cited by applicant]
US 20230019267A1 · Hapes et al. · 2023 [cited by applicant]
US 20230033451A1 · Reisinger et al. · 2023 [cited by applicant]
US 20230062568A1 · Temme et al. · 2023 [cited by applicant]
US 20230148607A1 · Rezaei et al. · 2023 [cited by applicant]
US 20230175959A1 · Wood et al. · 2023 [cited by applicant]
US 20230257317A1 · Temme et al. · 2023 [cited by applicant]
US 20230276807A1 · Reisinger et al. · 2023 [cited by applicant]
US 20230295559A1 · Ozaydin Eskiyenenturk et al. · 2023 [cited by applicant]
US 20240010576A1 · Temme et al. · 2024 [cited by applicant]
US 20240196903A1 · Reisinger et al. · 2024 [cited by applicant]
US 20240294953A1 · Eskiyenenturk et al. · 2024 [cited by applicant]
US 20240298647A1 · Reisinger et al. · 2024 [cited by applicant]
US 20240327851A1 · Tamsir et al. · 2024 [cited by applicant]
US 20240397955A1 · Belcher et al. · 2024 [cited by applicant]
US 20250027228A1 · Zhao et al. · 2025 [cited by applicant]
US 20250075222A1 · Bloch et al. · 2025 [cited by applicant]
US 20250115529A1 · Tamsir et al. · 2025 [cited by applicant]
US 20250137985A1 · Nevins et al. · 2025 [cited by applicant]
US 20250145544A1 · Strobel et al. · 2025 [cited by applicant]
US 20250185667A1 · Reisinger et al. · 2025 [cited by applicant]
US 20250207141A1 · Mirsky et al. · 2025 [cited by applicant]
US 20250228153A1 · O'Bryan et al. · 2025 [cited by applicant]
US 20250257316A1 · Voigt et al. · 2025 [cited by applicant]
US 20250346911A1 · Higgins et al. · 2025 [cited by applicant]
US 20250366419A1 · Bloch et al. · 2025 [cited by applicant]
US 20250369878A1 · Wood et al. · 2025 [cited by applicant]
US 20260015620A1 · Mirsky et al. · 2026 [cited by applicant]
US 20260041099A1 · Reisinger et al. · 2026 [cited by applicant]
US 20260041100A1 · Rezaei et al. · 2026 [cited by applicant]
BR 112020026676A2 · 2021 [cited by applicant]
CA 2051071A1 · 1993 [cited by applicant]
CN 101774854A · 2010 [cited by applicant]
CN 101899430A · 2010 [cited by applicant]
CN 102747002A · 2012 [cited by applicant]
CN 102770543A · 2012 [cited by applicant]
CN 103183572A · 2013 [cited by applicant]
CN 106086042A · 2016 [cited by applicant]
CN 108027849A · 2018 [cited by applicant]
CN 108271339A · 2018 [cited by applicant]
CN 108602729A · 2018 [cited by applicant]
FR 2494297 · 1982 [cited by applicant]
FR 2910230A1 · 2008 [cited by applicant]
JP 2018507708A · 2018 [cited by applicant]
RU 2511299C2 · 2014 [cited by applicant]
RU 2588473C1 · 2016 [cited by applicant]
WO WO8704182A1 · 1987 [cited by applicant]
WO WO9305154A1 · 1993 [cited by applicant]
WO WO9320685A1 · 1993 [cited by applicant]
WO WO9801088A1 · 1998 [cited by applicant]
WO WO9909834A2 · 1999 [cited by applicant]
WO WO0057183A1 · 2000 [cited by applicant]
WO WO0107567A1 · 2001 [cited by applicant]
WO WO03089640A2 · 2003 [cited by applicant]
WO WO2004074462A2 · 2004 [cited by applicant]
WO WO2005021585A2 · 2005 [cited by applicant]
WO WO2005038032A1 · 2005 [cited by applicant]
WO WO2006005100A1 · 2006 [cited by applicant]
WO WO2006083891A2 · 2006 [cited by applicant]
WO WO2006119457A1 · 2006 [cited by applicant]
WO WO2007027776A2 · 2007 [cited by applicant]
WO WO2009060012A2 · 2009 [cited by applicant]
WO WO2009091557A1 · 2009 [cited by applicant]
WO WO2011099019A1 · 2011 [cited by applicant]
WO WO2011099024A1 · 2011 [cited by applicant]
WO WO2011103247A2 · 2011 [cited by applicant]
WO WO2011103248A2 · 2011 [cited by applicant]
WO WO2011154960A1 · 2011 [cited by applicant]
WO WO2012139004A2 · 2012 [cited by applicant]
WO WO2012174271A2 · 2012 [cited by applicant]
WO WO2013076687A2 · 2013 [cited by applicant]
WO WO2013090628A1 · 2013 [cited by applicant]
WO WO2013132518A1 · 2013 [cited by applicant]
WO WO2013148290A1 · 2013 [cited by applicant]
WO WO2014042517A2 · 2014 [cited by applicant]
WO WO2014071182A1 · 2014 [cited by applicant]
WO WO2014078647A1 · 2014 [cited by applicant]
WO WO2014121366A1 · 2014 [cited by applicant]
WO WO2014201044A2 · 2014 [cited by applicant]
WO WO2016016629A1 · 2016 [cited by applicant]
WO WO2016016630A1 · 2016 [cited by applicant]
WO WO2016100727A1 · 2016 [cited by applicant]
WO WO2016108976A1 · 2016 [cited by applicant]
WO WO2016146955A1 · 2016 [cited by applicant]
WO WO2016178580A2 · 2016 [cited by applicant]
WO WO2016179046A1 · 2016 [cited by applicant]
WO WO2016181228A2 · 2016 [cited by applicant]
WO WO2016191828A1 · 2016 [cited by applicant]
WO WO2016200987A1 · 2016 [cited by applicant]
WO WO2017011602A1 · 2017 [cited by applicant]
WO WO2017042833A1 · 2017 [cited by applicant]
WO WO2017062412A1 · 2017 [cited by applicant]
WO WO2017069717A1 · 2017 [cited by applicant]
WO WO2018132774A1 · 2018 [cited by applicant]
WO WO2019032926A1 · 2019 [cited by applicant]
WO WO2019084059A2 · 2019 [cited by applicant]
WO WO2019084342A1 · 2019 [cited by applicant]
WO WO2019140125A1 · 2019 [cited by applicant]
WO WO2020006064A2 · 2020 [cited by applicant]
WO WO2020006246A1 · 2020 [cited by applicant]
WO WO2020014498A1 · 2020 [cited by applicant]
WO WO2020061363A1 · 2020 [cited by applicant]
WO WO2020092940A1 · 2020 [cited by applicant]
WO WO2020118111A1 · 2020 [cited by applicant]
WO WO2020132632A2 · 2020 [cited by applicant]
WO WO2020146372A1 · 2020 [cited by applicant]
WO WO2020163251A1 · 2020 [cited by applicant]
WO WO2020190363A1 · 2020 [cited by applicant]
WO WO2020191201A1 · 2020 [cited by applicant]
WO WO2020219893A2 · 2020 [cited by applicant]
WO WO2020219932A1 · 2020 [cited by applicant]
WO WO2021113352A1 · 2021 [cited by applicant]
WO WO2021146209A1 · 2021 [cited by applicant]
WO WO2021221689A1 · 2021 [cited by applicant]
WO WO2021221690A1 · 2021 [cited by applicant]
WO WO2021222567A2 · 2021 [cited by applicant]
WO WO2021222643A1 · 2021 [cited by applicant]
WO WO2021231449A2 · 2021 [cited by applicant]
WO WO2022029661A1 · 2022 [cited by applicant]
WO WO2022029661A9 · 2022 [cited by applicant]
WO WO2022140656A1 · 2022 [cited by applicant]
WO WO2022260676A1 · 2022 [cited by applicant]
WO WO2022261433A1 · 2022 [cited by applicant]
WO WO2023278804A1 · 2023 [cited by applicant]
WO WO2023147050A1 · 2023 [cited by applicant]
WO WO2023154805A2 · 2023 [cited by applicant]
WO WO2024006524A1 · 2024 [cited by applicant]
WO WO2024015230A1 · 2024 [cited by applicant]
WO WO2024137259A1 · 2024 [cited by applicant]
WO WO2025024750A2 · 2025 [cited by applicant]
US 8,476,226 B2, 07/2013, Lira et al. (withdrawn) [cited by applicant]
AU Patent Application No. 2019302747, Notice of Acceptance dated Nov. 5, 2025; Applicant Pivot Bio, Inc.; 3 total pages. [cited by applicant]
Australian Patent Application No. 2020218745, Office Action dated Sep. 2, 2025; Applicant Pivot Bio, Inc.; 7 total pages. [cited by applicant]
Connolly, L. N., et al., “Nitrogen fixation rates and aerial root production among maize landraces,” Frontiers in Plant Science 16: 1502884, pp. 1-15, Published Jan. 28, 2025, doi: 10.3389/fpls.2025.1502884. [cited by applicant]
Dumigan et al., “Ancient Relatives of Modern Maize from the Center of Maize Domestication and Diversification Host Endophytic Bacteria That Confer Tolerance to Nitrogen Starvation,” Frontiers.in Plant Science, Sep. 2021… [cited by applicant]
JP Patent Application No. 2025-016032, Office Action dated Dec. 2, 2025; Applicant Pivot Bio, Inc.; 4 total pages. [cited by applicant]
KR Patent Application No. 10-2020-7027921, Office Action mailed Oct. 31, 2025; Applicant Pivot Bio, Inc.; 8 total pages. [cited by applicant]
Rios-Galicia, B., et al. The Mexican giant maize of Jala landrace harbour plant-growth-promoting rhizospheric and endophytic bacteria, 3 Biotech 2021, 11 :447, 16 pages, Published online Sep. 24, 2021, doi: 10.1007/s132… [cited by applicant]
Rizo, J., et al., “Nitrogen Fixation in Pozol, a Traditional Fermented Beverage,” Applied and Environmental Microbiology, Aug. 2020, vol. 86, Issue 16, e00588-20, pp. 1-15. [cited by applicant]
U.S. Appl. No. 17/392,837, Final Office Action dated Nov. 18, 2025; Inventor Reisinger, Mark et al.; 8 pages. [cited by applicant]
U.S. Appl. No. 18/020,040, Non-Final Office Action dated Oct. 23, 2025; Inventor Reisinger, Mark et al.; 39 pages. [cited by applicant]
Van Deynze, A., et al., “Nitrogen fixation in a landrace of maize is supported by a mucilage-associated diazotrophic microbiota,” PLoS Biology 16(8):e2006352, pp. 1-21, Published Aug. 7, 2018, doi: 10.1371/journal.pbio.… [cited by applicant]
Wolf, E. S. A., et al., “Identification of genetic and environmental factors influencing aerial root traits that support biological nitrogen fixation in sorghum,” G3:Genes—Genomes—Genetics, 2024, 14(3), jkad285, 16 page… [cited by applicant]
Bageshwar, U. K., “Studies on Some Nitrogen Fixing Genes of Azotobacter Vinelandii,” Thesis submitted to the Jamia Millia Islamia for the award of Degree of Doctor of Philosophy, Department of Biosciences, Faculty of Na… [cited by applicant]
Bali, A. et al., “Excretion of ammonium by a nifL mutant of Azotobacter vinelandii fixing nitrogen,” Appl Environ Microbiol, 58(5):1711-1718 (1992); doi: 10.1128/aem.58.5.1711-1718.1992. [cited by applicant]
Barney et al., “Gene deletions resulting in increased nitrogen release by azotobacter vinelandii: application of a novel nitrogen biosensor,” Applied and Environmental Microbiology, Jul. 2015, 81(13), pp. 4316-4328. Pub… [cited by applicant]
Baum, J. A. et al., “Control of coleopteran insect pests through RNA interference,” Nature Biotechnology, 25(11):1322-1326 (2007). [cited by applicant]
Blanco, G. et al., “Sequence and molecular analysis of the nifL gene of Azotobacter vinelandii,” Mol Microbiol, 9(4):869-879 (1993); doi: 10.1111/j.1365-2958.1993.tb01745.x. [cited by applicant]
Bosmans, F. & Tytgat, J., “Sea anemone venom as a source of insecticidal peptides acting on voltage-gated Na+ channels,” Toxicon, 49(4):550-560 (2007). [cited by applicant]
Butzen, S., “Corn Seeding Rate Considerations,” Mar. 2019, 4 pages; www.pioneer.com/home/site/us/agronomy/library/corn-seeding-rate-considerations/. [cited by applicant]
CERA. “GM Crop Database. Center for Environmental Risk Assessment (CERA)”, ILSI Research Foundation, at cera-gmc.org/index.php?action=gm_crop_database, 1 page (2010). [cited by applicant]
Chakroun, M. et al., “Bacterial Vegetative Insecticidal Proteins (Vip) from Entomopathogenic Bacteria,” Microbiology and Molecular Biology Reviews, 80(2):329-350 (2016). [cited by applicant]
Chaurasia, A. K., et al., “Improved Eco-Friendly Recombinant [cited by applicant]
Chiang, L. W. et al., “Mutagenic Oligonucleotide-directed PCR Amplification (Mod-PCR): An Efficient Method for Generating Random Base Substitution Mutations in a DNA Sequence Element,” PCR Methods and Applications, 2(3)… [cited by applicant]
Ciampitti, I. A., et al., “Physiological perspectives of changes over time in maize yield dependency on nitrogen uptake and associated nitrogen efficiencies: A review,” Field Crops Research 133, pp. 48-67, doi:10.1016/j… [cited by applicant]
Clancy et al., “The domains carrying the opposing activities in adenylyl transferase are separated by a central regulatory domain,” FEBS Journal (2007), 274:2865-2877. [cited by applicant]
Communication pursuant to Article 94(3) EPC, mailed Feb. 16, 2023, for European Patent Application No. 19833252.0 (6 total pages). [cited by applicant]
Compant, S., et al., “Plant growth-promoting bacteria in the rhizo- and endosphere of plants: Their role, colonization, mechanisms involved and prospects for utilization,” Soil Biology and Biochemistry (2010), 42(5), pp… [cited by applicant]
Crickmore, et al. “Bacillus thuringiensis toxin nomenclature,” 2016, Web page, https://web.archive.org/web/20161110143346/http://www.btnomenclature.info/, 2016, 1 page, retrieved from internet on Nov. 30, 2022. [cited by applicant]
Crickmore, N. et al., “Revision of the Nomenclature for the Bacillus thuringiensis Pesticidal Crystal Proteins,” Microbiol. Mol. Biol. Rev., 62(3):807-813 (1998). [cited by applicant]
Crickmore, N., The VIP nomenclature, http://www.lifesci.sussex.ac.uk/home/Neil_Crickmore/Bt/vip.html , retrieved from the internet Nov. 11, 2022 (4 total pages). [cited by applicant]
Danhorn, T., et al., “Biofilm formation by plant-associated bacteria,” Annual Review of Microbiology (2007), 61, 401-422, doi.org/10.1146/annurev.micro.61.080706.093316, 25 pages. [cited by applicant]
Dixon, R., & Kahn, D. (2004). Genetic regulation of biological nitrogen fixation. Nature Reviews. Microbiology, 2(8), 621-31. https://doi.org/10.1038/nrmicro954. [cited by applicant]
Examination Report, dated Jul. 31, 2023, for Canadian Patent Application No. 3,105,529 (5 pages). [cited by applicant]
Extended European Search Report and Search Opinion, dated Jul. 30, 2024, for European Patent Application No. 21854590.3, 7 pages. [cited by applicant]
Extended European Search Report and Search Opinion, dated Mar. 3, 2023, for European Application No. 20751885.3 (19 total pages). [cited by applicant]
Extended European Search Report, dated Mar. 14, 2022, for European Application No. 19833252.0 (6 total pages). [cited by applicant]
Eyraud, V. et al., “Expression and Biological Activity of the Cystine Knot Bioinsecticide PA1b (Pea Albumin 1 Subunit b),” PLoS One, 8(12): e81619 (2013), 9 pages; doi:10.1371/journal.pone.0081619. [cited by applicant]
Fausti, S. et al., “Cost of Crop Production,” Practical Mathematics for Precision Farming, Oct. 23, 2017, pp. 191-199, doi: 10.2134/practicalmath2017.0032. [cited by applicant]
First Examination Report, dated Sep. 6, 2022, for Indian Patent Application No. 202117002885 (5 total pages). [cited by applicant]
First Office Action, issued Jun. 22, 2022, for Chinese Patent Application No. 201980053956.9, with English translation, 21 pages. [cited by applicant]
First Office Action, issued May 27, 2022, for Chinese Patent Application No. 202080027155.8, with English translation, 16 total pages. [cited by applicant]
Gaby, J. C. & Buckley, D. H., “A comprehensive aligned nifH gene database: a multipurpose tool for studies of nitrogen-fixing bacteria,” Database, vol. 2014: article ID bau001, 8 pages (2014). doi:10.1093/database/bau00… [cited by applicant]
Glick, B. R. et al., “Plant Growth-Promoting Bacteria: Mechanisms and Applications,” Scientifica, Oct. 2012, vol. 2012, ID 963401, 15 pages, https://doi.org/10.6064/2012/963401. [cited by applicant]
Hearing Notice for Indian Patent Application No. IN202117002885 dated Jan. 10, 2024, 2 pages. [cited by applicant]
Hidaka, M. et al., “Promotion of the Growth of Rice by Inoculation of Nitrogen-Fixing-Activity-Enhanced Bacteria to the Rhizosphere,” Nitrogen Fixation: From Molecules to Crop Productivity, p. 445 (Jan. 1, 2002). [cited by applicant]
Hoeschle-Zeledon, I. et al., “Regulatory challenges for biological control,” The CGIAR Systemwide Program on Integrated Pest Management, Jan. 2013, SP-IPM Secretariat, International Institute of Tropical Agriculture (II… [cited by applicant]
International Preliminary Report on Patentability for International Application No. PCT/IB2021/057171 dated Feb. 16, 2023, 8 pages. [cited by applicant]
International Preliminary Report on Patentability, mailed Aug. 19, 2021, for International Application No. PCT/US2020/016471 (13 total pages). [cited by applicant]
International Preliminary Report on Patentability, mailed Jan. 21, 2021, for International Application No. PCT/US2019/041429 (12 total pages). [cited by applicant]
International Search Report and Written Opinion for Patent Application No. PCT/IB2021/057171, mailed Nov. 5, 2021, 13 pages. [cited by applicant]