DETAILED ACTION
Notice of Pre-AIA or AIA Status
The present application, filed on or after March 16, 2013, is being examined under the first inventor to file provisions of the AIA .
Response to Previous Communication
In response to the restriction requirement mailed on 04/13/2026, the Applicant’s attorney, Xiang Li, elects Group 1 (i.e. Claims 1-18 and 20-22) without traversal which was filed on 07/09/2026. Applicant further elects the nucleotide sequence SEQ ID NO:135, and the corresponding protein sequence SEQ ID NO: 1 for IMA, and the nucleotide sequence SEQ ID NO: 152, and the corresponding protein sequence SEQ ID NO: 139 for BTSL.
The examiner acknowledges the amendments to the claims filed on 07/09/2026.
Priority
This application claims priority to provisional application 63/601,364 filed on 11/21/2023.
Status of the Claims
Claims 1-23 are pending.
Claims 19 and 23 are withdrawn from consideration for being directed to non-elected invention(s).
Claims 1-18 and 20-22 are examined herein.
Claim Interpretation
IMA1 is an identical protein to FEP3 (AT1G47400) and the names can be used interchangeably (see Lichtblau, Daniela M., et al. "The iron deficiency-regulated small protein effector FEP3/IRON MAN1 modulates interaction of BRUTUS-LIKE1 with bHLH subgroup IVc and POPEYE transcription factors." Frontiers in Plant Science 13 (2022): 930049 page 10 paragraph 4).
Drawings
The examiner believed there was an error in the labels in Fig. 2B which should be corrected so flg22 was added in both Fe+ and Fe- conditions.
Claim Objection
Claims 20-22 are objected to for referencing withdrawn Claim 19.
Improper Markush Group
Claims 10 and 17 are rejected under the judicially-created basis that they contain an improper Markush groupings of alternative species. See In re Harnisch, 631 F.2d 716, 721-722 (CCPA 1980) and Ex parte Hozumi, 3 USPQ2d 1059, 1060 (Bd. Pat. App. and Int. 1984). The improper Markush groupings includes species of the claimed invention that do not share both a substantial structural feature and a common use that flows from the substantial structural feature. All dependent claims are included in these rejections unless they contain a limitation that overcomes the deficiencies of the parent claim from which they depend.
The members of the improper Markush groupings do not share a substantial feature and a common use that flows from the substantial structural feature for the following reasons: the claims encompass thousands of unique polypeptide sequences (and the corresponding thousands of unique nucleic acid sequences), which are biochemically divergent, they have no conserved structure throughout the genus other than a phosphodiester backbone or a polypeptide backbone (peptide linkages), respectively, and may or may not be associated with conveying resistance against a plant pathogen in a plant.
The species recited in Claim 10 lack a substantial structural feature for the claimed 80% amino acid similarity to SEQ ID NO: 1 or nucleotide sequence similarity to SEQ ID NO: 135 as evidenced by the failure of the non-elected sequences (i.e for example, SEQ ID NOs:137 and 98) to claim a sequence with similarity higher than the claimed 80% similarity to the elected sequence, SEQ ID NOs:135 and 1, respectively.
The species recited in Claim 17 lack a substantial structural feature for the claimed 80% amino acid similarity to SEQ ID NO: 139 or nucleotide sequence similarity to SEQ ID NO: 152 as evidenced by the failure of the non-elected sequences (for example, SEQ ID NOs: 153 and 145) to claim a sequence with similarity higher than the claimed 80% similarity to the elected sequence, SEQ ID NOs:152 and 139, respectively.
Claim Rejections - 35 USC § 112(b)
The following is a quotation of 35 U.S.C. 112(b):
(b) CONCLUSION.—The specification shall conclude with one or more claims particularly pointing out and distinctly claiming the subject matter which the inventor or a joint inventor regards as the invention.
The following is a quotation of 35 U.S.C. 112 (pre-AIA ), second paragraph:
The specification shall conclude with one or more claims particularly pointing out and distinctly claiming the subject matter which the applicant regards as his invention.
Claim 2 is rejected under 35 U.S.C. 112(b) or 35 U.S.C. 112 (pre-AIA ), second paragraph, as being indefinite for failing to particularly point out and distinctly claim the subject matter which the inventor or a joint inventor (or for applications subject to pre-AIA 35 U.S.C. 112, the applicant), regards as the invention.
Claims 2 recites a “foliar bacterial pathogen” which is not clearly defined in the disclosure. It is unclear if this is a pathogen that only infects leaves, can colonize leaves but does not infect through the leaf, or symptoms of infection modifies the leaves in some way.
Claim Rejections - 35 USC § 112
The following is a quotation of the first paragraph of 35 U.S.C. 112(a):
(a) IN GENERAL.—The specification shall contain a written description of the invention, and of the manner and process of making and using it, in such full, clear, concise, and exact terms as to enable any person skilled in the art to which it pertains, or with which it is most nearly connected, to make and use the same, and shall set forth the best mode contemplated by the inventor or joint inventor of carrying out the invention.
The following is a quotation of the first paragraph of pre-AIA 35 U.S.C. 112:
The specification shall contain a written description of the invention, and of the manner and process of making and using it, in such full, clear, concise, and exact terms as to enable any person skilled in the art to which it pertains, or with which it is most nearly connected, to make and use the same, and shall set forth the best mode contemplated by the inventor of carrying out his invention.
Written Description
Claims 1-18 and 20-22 are rejected under 35 USC § U.S.C. 112(a) or 35 U.S.C. 112 (pre-AIA ), first paragraph, as failing to comply with the written description requirement. The claim(s) contains subject matter which was not described in the specification in such a way as to reasonably convey to one skilled in the relevant art that the inventor or a joint inventor, or for applications subject to pre-AIA 35 U.S.C. 112, the inventor(s), at the time the application was filed, had possession of the claimed invention.
The Federal Circuit has clarified the written description requirement. The court stated that a written description of an invention "requires a precise definition, such as by structure, formula, [or] chemical name, of the claimed subject matter sufficient to distinguish it from other materials". University of California v. Eli Lilly and Co., 119 F.3d 1559, 1568; 43 USPQ2d 1398, 1406 (Fed. Cir. 1997). The court also concluded that "naming a type of material generally known to exist, in the absence of knowledge as to what that material consists of, is not description of that material". Id. Further, the court held that to adequately describe a claimed genus, Patent Owner must describe a representative number of the species of the claimed genus, and that one of skill in the art should be able to "visualize or recognize the identity of the members of the genus".
The claims are broadly drawn to a method for increasing resistance against a plant pathogen by increasing expression and/or activity of Iron Man (IMA) or reducing expression and/or activity of BRUTUS-Like E3 Ligase (BTSL).
Applicant describes:
Arabidopsis treated with flg22 (elicitor peptide fragment of bacterial flagellin) repress iron uptake during iron deficiency and IMA1 is responsible for this process as overexpressing IMA1 prevents this phenotype. BTSL1&2 are required for degrading IMAI protein and the btsl 1,2 double mutant are less sensitive to the flg22-triggered repression of iron deficiency responses.
Compared to Col-0, the btsl 1,2 mutant plants developed less leaf chlorosis in response to flg22 under low iron conditions, suggesting that btsl 1,2 plants are less sensitive to the flg22-triggered repression of iron deficiency responses (Figs. 4A-4C).
By analyzing IMA1ox dependent transcriptome changes in the shoot from a published dataset, it was found that besides iron-responsive genes, immune response and systemic acquired resistance genes were enriched among the upregulated genes.
UBQ10::mCitrine-IMA1 plants were more resistant to foliar bacterial pathogen Pseudomonas syringae pv. Tomato DC3000.
Locally gated MAMP responsiveness can lead to spatially confined IMA1 repression and thereby might contribute to allow roots to locally shut off iron deficiency responses when internally colonized or to continue with iron acquisition in the presence of non-invasive, surface dwelling bacteria.
Applicant does not describe:
Any other plant, besides Arabidopsis overexpressing IMA1, that is resistant to a foliar bacterial pathogen (i.e. Pseudomonas syringae, Ralswnia solanacearum, Xanthomonas myzae, Xanthomonas campestris, Xanthomonas axonopodis, Erwinia amylovora, Xylella fastidiosa, Dickeyo dadantii, or Pectobacteriwn carotovorwn).
All plants with increased activity or expression of an IMA or IMA1 genes (i.e. SEQ ID NO: 135) or protein/c-terminal fragment (i.e. SEQ ID NO:209) that is resistant to all plant pathogens.
All plants with reduced expression or activity of a BTSL genes (i.e. SEQ ID NO: 152) or protein (i.e. SEQ ID NO:139) that is resistant to all plant pathogens.
All sequences with 80% similarity to SEQ ID NOs: 1, 135, 139, or 152 which can be up or down regulated to convey resistance to all plant pathogens.
Claims 1-18 and 20-22 encompass a vast genus of polynucleotide and polypeptide sequences. With the exception of sequences in the specification, the Applicants have failed to provide working examples of all possible gene and protein sequences for IMA or BTSL or sequences with same function and have at least 80% similarity with SEQ ID NOs: 1, 135, 139, or 152 which will confer the phenotype of foliar bacterial pathogen (i.e. Pseudomonas syringae, Ralswnia solanacearum, Xanthomonas myzae, Xanthomonas campestris, Xanthomonas axonopodis, Erwinia amylovora, Xylella fastidiosa, Dickeyo dadantii, or Pectobacteriwn carotovorwn) resistance in all possible plants as drawn to in Claims 1-17 and 20-22. Looking at the shortest sequence (i.e. SEQ ID NO: 1) as an example, describing and reducing to practice a genus of proteins with all possible single amino acid substitutions relative to the 50 amino acid long polypeptide of SEQ ID NO: 1 would require describing and reducing to practice 2050 amino acid sequences. Protein consisting of a sequence 80% identical to SEQ ID NO:1 would have up to ~10 random amino acid substitutions relative to SEQ ID NO: 1. Accordingly, 10 x 2050 amino acid sequences would need to be described and reduced to practice; most of which were not in Applicants’ possession at the time of filing. The instant Specification fails to provide guidance for which exact nucleotides could be mutated with the exception of the conserved C-terminal fragment. More importantly, all C-terminal fragment sequences (as was recited in Claims 6-8, and 10) are also not described and could be interpreted to mean any number, type, and length of sequence(s). Furthermore, Zhao, Junhui, et al. "Arabidopsis BRUTUS, BRUTUS-LIKE, and bHLH IVc subgroup proteins coordinate iron homeostasis in the root and shoot." The Plant Cell 38.2 (2026): koag006 teaches that BTSL protein and genes are only found in dicotyledon plants on page 13 paragraph 2. These factors do not appear to have been adequately addressed in the instant application.
Claims 1-3, 5-18 and 20-22 also encompass a vast number of plant pathogens that were not disclosed by the applicant but could be granted resistance to upon increased expression of IMA or reduced expression of BTSL. Buoso, Sara, et al. "‘Candidatus Phytoplasma solani’ interferes with the distribution and uptake of iron in tomato." BMC genomics 20.1 (2019): 703 teaches a plant pathogen, Candidatus Phytoplasma solani, that was not in the applicant’s disclosure. It infects many plants and “Typical symptoms of infected plants include yellowing of leaves [i.e. foliar of Claim 2] or shoots [i.e. Claim 3], leaf curling, and general stunting, but the molecular mechanisms underlying most of the reported changes remain largely enigmatic”.
It is also known in the art that some common plants are recalcitrant to transformation. For example, Claims 1-17 and 20-22 are directed to modifying all plants. However, the abstract of Guo, Guanghui, et al. "An elegant co‐transformation strategy for recalcitrant wheat using morphogenic regulators." The Plant Journal 124.2 (2025): e70541 teaches wheat transformation efficiency largely depends on the recipient genotype and overcoming low regeneration efficiency is a barrier to successfully transforming wheat. Morphogenic regulators are shown to improve wheat transformation and regeneration, however, there are still issues regarding how to specifically utilize them in recalcitrant wheat. These challenges are described on page 2 in paragraphs 1-2 and solutions for overcoming these barriers were not provided in the disclosure. Claim 18 specifically recites the limitation of using a soybean plant. Xu, Hu, et al. "Progress in soybean genetic transformation over the last decade." Frontiers in plant science 13 (2022): 900318 teaches “Although much effort has been made to improve the transformation systems for soybean, there are some challenges such as genotype flexibility, low transformation frequency, time to time chimerism in T0 transgenic plants, and availability of a system for new breeding technologies such as genome editing”. Overcoming these challenges was not addressed in the specification and would require substantial trial and error testing.
All possible sequences with 80% identity to SEQ ID NOs: 1, 135, 139, or 152 and C-terminal sequences of IMA proteins which grants resistance as well as a description of which plant pathogens in which plants these methods can be used in were not described. Therefore, given the lack of written description in the instant disclosure with regard to the structural and functional characteristics of the claimed compositions, Applicant does not appear to have been in possession of the claimed genus at the time this application was filed.
Examiner’s Note: Applicant is encouraged to specifically indicate specific genetic modifications which would confer resistance to a specific pathogen and have evidence in support for it.
Scope of Enablement
Claims 1-18 and 20-22 are rejected under 35 USC § U.S.C. 112(a) or 35 U.S.C. 112 (pre-AIA ), first paragraph, because the specification, while being enabling for granting Arabidopsis, transformed with the UBQ10::mCitrine-IMA1 construct, resistance to the foliar bacterial pathogen Pseudomonas syringae pv. Tomato DC3000 does not enable any person skilled in the art to which it pertains, or with which it is most nearly connected, to make or use the invention commensurate in scope with these claims.
In re Wands lists a number of factors for determining whether or not undue experimentation would be required by one skilled in the art to make and/or use the invention. These factors are: (1) the quantity of experimentation necessary; (2) the amount of direction or guidance presented; (3) the presence or absence of working examples of the invention; (4) the nature of the invention; (5) the state of the prior art; (6) the relative skill of those in the art; (7) the predictability or unpredictability of the art; (8) the breadth of the claim. In re Wands, 858 F.2d 731, 8 USPQ2d 1400 (Fed. Cir. 1988).
The claims are broadly drawn to a method for increasing resistance against a plant pathogen by increasing expression and/or activity of Iron Man (IMA) or reducing expression and/or activity of BRUTUS-Like E3 Ligase (BTSL).
Applicant Teaches:
Arabidopsis treated with flg22 (elicitor peptide fragment of bacterial flagellin) repress iron uptake during iron deficiency and IMA1 is responsible for this process as overexpressing IMA1 prevents this phenotype. BTSL1&2 are required for degrading IMAI protein and the btsl 1,2 double mutant are less sensitive to the flg22-triggered repression of iron deficiency responses.
Compared to Col-0, the btsl 1,2 mutant plants developed less leaf chlorosis in response to flg22 under low iron conditions, suggesting that btsl 1,2 plants are less sensitive to the flg22-triggered repression of iron deficiency responses (Figs. 4A-4C).
By analyzing IMA1ox dependent transcriptome changes in the shoot from a published dataset, it was found that besides iron-responsive genes, immune response and systemic acquired resistance genes were enriched among the upregulated genes.
UBQ10::mCitrine-IMA1 plants were more resistant to foliar bacterial pathogen Pseudomonas syringae pv. Tomato DC3000.
Locally gated MAMP responsiveness can lead to spatially confined IMA1 repression and thereby might contribute to allow roots to locally shut off iron deficiency responses when internally colonized or to continue with iron acquisition in the presence of non-invasive, surface dwelling bacteria.
Applicant Does Not Teach:
Any other plant, besides Arabidopsis overexpressing IMA1, that is resistant to a foliar bacterial pathogen (i.e. Pseudomonas syringae, Ralswnia solanacearum, Xanthomonas myzae, Xanthomonas campestris, Xanthomonas axonopodis, Erwinia amylovora, Xylella fastidiosa, Dickeyo dadantii, or Pectobacteriwn carotovorwn).
All plants with increased activity or expression of an IMA or IMA1 genes (i.e. SEQ ID NO: 135) or protein/c-terminal fragment (i.e. SEQ ID NO:209) that is resistant to all plant pathogens.
All plants with reduced expression or activity of a BTSL genes (i.e. SEQ ID NO: 152) or protein (i.e. SEQ ID NO:139) that is resistant to all plant pathogens.
All sequences with 80% similarity to SEQ ID NOs: 1, 135, 139, or 152 which can be up or down regulated to convey resistance to all plant pathogens.
The instant disclosure and the instant claims do not set forth structural characteristics essential to increasing expression and or activity of IMA or decreasing expression or activity of BTSL in all plants for increased resistance against all plant pathogens.
Claims 1, 5-17 and 20-22 are drawn to increasing resistance to plant pathogens in all plants while Claims 2-3 limit the pathogens to foliar bacterial pathogens and/or that infects the shoot of the plant, Claim 4 limits the pathogen species to Pseudomonas syringae, Ralswnia solanacearum, Xanthomonas myzae, Xanthomonas campestris, Xanthomonas axonopodis, Erwinia amylovora, Xylella fastidiosa, Dickeyo dadantii, or Pectobacteriwn carotovorwn and Claim 18 limits the plant species to pennycress, soybean, canola, rice, tomato, Zea mays, or sorghum. Wang, Peilin, et al. "Plant Genetic Engineering: Technological Pathways, Application Scenarios, and Future Directions." Advanced Science 13.30 (2026): e21040 teaches extensive experimentation is needed to determine if plants harboring a single resistance gene actually demonstrates resistance in the field. This is taught on page 8 paragraph 4 which recites “under laboratory or greenhouse conditions, plants harboring single resistance genes often exhibit nearly complete protection. However, when exposed to natural field environments, resistance frequently diminishes or collapses due to ecological and evolutionary complexities. To avoid overinterpreting single- gene yield claims, field translation should be evaluated against commonly recommended criteria, including multi-location and multi-year testing, appropriate elite genetic backgrounds and agronomic management, robust experimental design and statis- tics, and assessment of yield components and trade-offs under realistic production conditions”. This indicates the experiment with UBQ10::mCitrine-IMA1 plants may not translate to plants in the field for all plant pathogens.
The Applicant has provided only the working example for expressing the UBQ10::mCitrine-IMA1 construct in Arabidopsis for Pseudomonas syringae pv. Tomato DC3000 resistance, but not in any other plant species or pathogens. Thus, the examples provided by the Applicant do not provide adequate working examples to enable the scope of the invention without undue experimentation. Given the breadth of the claims, the lack of guidance and working examples, the unpredictability in the art, and the state of the art, undue experimentation would be required to make and use the claimed invention, and therefore, the invention is not enabled throughout the broad scope of the claims.
Claim Rejections - 35 USC § 102/103
A rejection under 35 U.S.C. § 102 or § 103 does not require the same analysis as a rejection under 35 U.S.C. § 102 or a rejection under 35 U.S.C. § 103. The rejection is made because the examiner cannot determine whether the prior art composition possesses characteristics that are not recited in the art. The examiner does not have sufficient facts to determine whether the claimed compositions, polynucleotides, polypeptides and organisms are inherently the same as the prior art compositions, polynucleotides, polypeptides and organisms. In addition, the examiner cannot conclude that the claimed subject matter would have been obvious since it cannot be determined whether the claimed and prior art compositions, polynucleotides, polypeptides and organisms differ. Where the prior art product seems to be identical, except that the prior art is silent to a characteristic or property claimed, then the burden shifts to applicants to provide evidence that the prior art would neither anticipate nor render obvious the claimed invention. In re Best, 195 USPQ 430, 433 (CCPA 1977).
Claims 1-6, 9-10, and 15-16 are rejected as being anticipated under 35 U.S.C. § 102(a)(1), or alternatively as being unpatentable under 35 U.S.C. § 103, over Lichtblau, Daniela M., et al. "The iron deficiency-regulated small protein effector FEP3/IRON MAN1 modulates interaction of BRUTUS-LIKE1 with bHLH subgroup IVc and POPEYE transcription factors." Frontiers in Plant Science 13 (2022): 930049 (see IDS filed 11/21/2024).
Claims 1-6, 9-10, and 15-16 are drawn to increasing expression of a full length IMA in a plant for increased pathogen resistance by introducing an exogenous nucleic acid molecule. The IMA protein and nucleotide sequences have 80% similarity to SEQ ID NOs:1 and 135 and is operably linked to a promoter that can be tissue specific.
Lichtblau et al 2022 is directed to protein interactions among 23 proteins of the –Fe response (see page 3 paragraph 4). The authors disclose the creation of a FEP3-OX (i.e. Iron Man 1(IMA1) Arabidopsis plant. Their expression can be seen in Supplementary Figure 9A and a description of how they were made is on page 16 paragraph 4. These described plants have increased expression and activity of IMA as was drawn to in Claim 1. The increase in expression was induced by transforming a plant with a vector harboring the HA-tagged FEP3/IMA1 (i.e. exogenous nucleic acid molecule that encodes a full length IMA protein of Claims 5-6) under control of a double CaMV35S promoter (i.e. operably linked as was recited in Claim 15) using Agrobacteria (Rhizobium radiobacter) strain GV3101. The IMA1 protein used was cloned from Arabidopsis and overexpressed in the native plant as was drawn to in Claim 9. Additionally, the gene overexpressed (i.e. exogenous nucleic acid molecule) appears to have 100% identity to applicant’s SEQ ID NOs:1 and 1351 as was recited in Claim 10 (see below). Regarding Claim 16, Lichtblau et al 2022 also teaches the generation of a proFEP3:FEP3-GUS construct that was transformed into Arabidopsis WT plants. This indicates the transformed wild type plants were also overexpressing IMA as they contained a second copy under the native FEP3 promoter. The FEP3 promoter also demonstrates tissue-specific activity in the vascular cells of the plant (see supplementary figure 6C and 6I).
100.0% identity in 50 residues overlap; Score: 257.0; Gap frequency: 0.0%
Sequence1 1 MMSFVANLAIKRFDHASTVYVEDVVDSSRVAYSENGGDDDDSGYDYAPAA
Sequence2 1 MMSFVANLAIKRFDHASTVYVEDVVDSSRVAYSENGGDDDDSGYDYAPAA
**************************************************
Sequence1: Applicant’s SEQ ID NO:1
Sequence2: Lichtblau et al 2022 sequence for A.thaliana_AT1G47400 Supplementary Figure 7
Accordingly, Lichtblau et al 2022 anticipates the claimed invention.
Lichtblau et al 2022 do not explicitly teach their plants are resistant against a foliar bacterial plant pathogen that infects a shoot of the plant and can be Pseudomonas syringae, Ralswnia solanacearum, Xanthomonas myzae, Xanthomonas campestris, Xanthomonas axonopodis, Erwinia amylovora, Xylella fastidiosa, Dickeyo dadantii, or Pectobacteriwn carotovorwn (i.e. Claims 1-4); however, this property is interopreted as being inherent to the compositions and methods taught by Lichtblau et al 2022. Even if it was determined that Lichtblau et al 2022 does not explicitly teach each of the limitations in the instant claims, the claims are drawn to a method that results in resistance to pathogens. The method active step (i.e. overexpressing IMA) has been previously demonstrated by Lichtblau et al 2022. This indicates that the plants overexpressing IMA in Lichtblau et al 2022 are also resistant to plant pathogens (absent evidence to the contrary).
Claims 1, 5-8, 18 and 20 are rejected as being anticipated under 35 U.S.C. § 102(a)(1), or alternatively as being unpatentable under 35 U.S.C. § 103, over US 20150315250 A1 (Schmidt).
Claims 1, 5-8, 18 and 20 are drawn to increasing expression and/or activity of the C-terminal fragment of IMA in a plant by introducing one or more exogenous nucleic acid molecules for plant pathogen resistance. The C-terminal fragment can be made up of 7-20 amino acids and have the amino acid sequence of SEQ ID NO: 209. The IMA C-terminal fragment can have increased expression in soybean (i.e. Claim 18) while Claim 20 is drawn to a method for producing a commodity plant product from the gene edited plant.
Schmidt is directed to producing transgenic plants transformed with a recombinant polynucleotide (i.e. exogenous nucleic acid molecules of instant Claim 5) encoding the IMA1 or IMA3 polypeptide which facilitate uptake and circulation of the trace elements into the plants (see abstract). Claims 1 and 3-5 of Schmidt are directed to a transgenic plant overexpresses (i.e. increase expression or activity as was drawn to in instant Claim 1) the iron-regulated polypeptide that “comprises a C-terminal motif comprising from N-terminal to C-terminal a first domain of GDDDD (SEQ ID NO: 1), and a second domain of DXAPAA (SEQ ID NO: 2)”. The peptide spacer can be 10 or less amino acid residues and the iron-regulated polypeptide comprises 20-100 amino acid residues in length. The iron-regulated polypeptide described by Claims 1 and 3-5 of Schmidt teach the limitations of instant Claims 6 (i.e. a C-terminal fragment of IMA) that is 20 amino acids long (i.e. instant Claim 7) that comprises the sequences indicated in Claim 8 (i.e. SEQ ID NO:209). Schmidt also teaches the application can be used for tomato (i.e. instant Claim 18) in paragraph 63 and shows a tomato plant overexpressing IMA1 in figure 4. Paragraph 84 teaches how the tomato and Arabidopsis transformants were generated. The transformed tomato fruit in Figure 4b also indicate Schmidt was in possession of a method that produce a commodity plant product from a gene-edited plant with increased expression of IMA (i.e. Claim 20).
Accordingly, Schmidt anticipates the claimed invention.
Even if it was determined that Schmidt does not explicitly teach each of the limitations in the instant claims (i.e. plants are resistant against a plant pathogen as was a limitation of Claim 1), the claims are drawn to a method that results in resistance to pathogens. The method active step (i.e. overexpressing IMA C-terminal fragment with SEQ ID NO: 209) has been previously described/claimed by Schmidt. This indicates that the plants overexpressing IMA or a C-terminal fragment in Schmidt are also resistant to foliar bacterial plant pathogen (absent evidence to the contrary).
Claims 1, 11-12, 17 and 21-22 are rejected as being anticipated under 35 U.S.C. § 102(a)(1), or alternatively as being unpatentable under 35 U.S.C. § 103, over Rodríguez-Celma, Jorge, et al. "Arabidopsis BRUTUS-LIKE E3 ligases negatively regulate iron uptake by targeting transcription factor FIT for recycling." Proceedings of the National Academy of Sciences 116.35 (2019): 17584-17591 as evidenced by NCBI Reference Sequence: NM_106135.3 (Theologis et al 2022).
Claims 1, 11-12, 17 and 21-22 are drawn to reducing activity and/or expression of BTSL for plant pathogen resistance by introducing one or more exogenous nucleic acid molecules into a plant that generates a deletion or loss-of-function mutation in the BTSL gene. Claims 21 is drawn to a method of producing plant seed by crossing the gene edited plant with itself or another plant. Claim 22 is drawn to a method of crossing the gene edited plant with another plant and selecting progeny with increased disease resistance.
Rodríguez-Celma et al 2019 is directed to describing “the function of 2 partially redundant E3 ubiquitinligases, BRUTUS-LIKE1 (BTSL1) and BTSL2, in Arabidopsis thaliana” (see abstract). It also recites “Two genes homologous to BTS are found in the Arabidopsis genome, named BRUTUS-LIKE1 (AT1G74770) and BRUTUS-LIKE2 (AT1G18910) on page 2 paragraph 1. The btsl1 btsl2 double knockout [i.e. reduced expression and/or activity of instant Claim 1] mutant accumulated Fe and was more tolerant to Fe deficiency”. Page 2 paragraph 1 of the materials and methods section of Rodríguez-Celma et al 2019 teaches multiple SALK lines with T-DNA insertion (i.e. introduced exogenous nucleic acid molecules that reduce expression of BTSL; Claim 11). The method used to generate double knockout mutants plants was described on page 3 paragraph 3 which recites “double knockout [i.e. loss-of-function of Claim 12] line was produced by crossing btsl1-1 with the btsl2-2 line”. Because the double knock out plants were generated by crossing two gene-edited plants with reduced BTSL activity together, the method of producing seeds for double knockout mutants teaches the limitations of Claim 21. Page 1 Paragraph 1 of the materials and methods section of Rodríguez-Celma et al 2019 also introduces the method for breeding a plant with increased disease resistance (i.e. Claim 22). The plants with the btsl mutations were crossed and the progeny (i.e. planted seeds from cross) were presumably genotyped (i.e. selected) using PCR. Selecting plants with the desired genotype would inherently select for plants with the phenotype of plant pathogen resistance. Finally, regarding Claim 17 which recites the limitations of elected sequences with SEQ ID NOs: 152 and 139. These sequences have been known in the art since 2022 and identified as “AT1G74770” as evidenced by NCBI Reference Sequence: NM_106135.3 (Theologis et al 2022) which teaches a DNA sequence and amino acid sequence with 100% similarity to Applicant’s SEQ ID NOs: 152 and 139 (see below).
Query: Applicant’s SEQ ID NO:152
Sbjct: DNA Sequence from Theologis et al 2022
Query 1 ATGGGAGGCGGAAATCTTCATTCCCTTCCGCCGGAAAACGCTTCCGTTTCTGCATCTTAT 60
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Sbjct 123 ATGGGAGGCGGAAATCTTCATTCCCTTCCGCCGGAAAACGCTTCCGTTTCTGCATCTTAT 182
Query 61 GCGGTGACCGTTGGAAATACCAAGCTTTCTGATGCTCCGGTTCTTTTCTTCGTTTACTGC 120
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Sbjct 183 GCGGTGACCGTTGGAAATACCAAGCTTTCTGATGCTCCGGTTCTTTTCTTCGTTTACTGC 242
Query 121 CACAAGGCTTTCCGTGCTCAGCTGGTGGAGCTCCGGCGTTTCGCTACTGACGCTGCTGAA 180
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Sbjct 243 CACAAGGCTTTCCGTGCTCAGCTGGTGGAGCTCCGGCGTTTCGCTACTGACGCTGCTGAA 302
Query 181 GCCGATTCCTTTAGTGGGGATCTGGCGGTGGAGCTCTCCCGGAAGTTTGAGTTCTTGAAG 240
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Sbjct 303 GCCGATTCCTTTAGTGGGGATCTGGCGGTGGAGCTCTCCCGGAAGTTTGAGTTCTTGAAG 362
Query 241 CTTGTATATAAGTATCATAGCGCAGCTGAAGATGAGGTTATATTTTTGGCGCTAGATAAA 300
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Sbjct 363 CTTGTATATAAGTATCATAGCGCAGCTGAAGATGAGGTTATATTTTTGGCGCTAGATAAA 422
Query 301 CGAGTGAAAAACATAGTTTCTAATTACTCACTTGAGCATGCTGGCACAGATGATCTCTTT 360
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Sbjct 423 CGAGTGAAAAACATAGTTTCTAATTACTCACTTGAGCATGCTGGCACAGATGATCTCTTT 482
Query 361 ACCTCAATTTTCCACTGGCTACATGTTCTTGAAGAAGAAATAGGAAGTAGAAGCGATGTA 420
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Sbjct 483 ACCTCAATTTTCCACTGGCTACATGTTCTTGAAGAAGAAATAGGAAGTAGAAGCGATGTA 542
Query 421 CTTCGTGAAGTTATTTTATGCATAGGCACCATCCAGTCATCTATATGCCAACATATGCTT 480
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Sbjct 543 CTTCGTGAAGTTATTTTATGCATAGGCACCATCCAGTCATCTATATGCCAACATATGCTT 602
Query 481 AAAGAAGAGCGGCAGGTGTTTCCTTTGTTAATAGAGAAGTTTTCTTTCCGAGAACAAGCA 540
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Sbjct 603 AAAGAAGAGCGGCAGGTGTTTCCTTTGTTAATAGAGAAGTTTTCTTTCCGAGAACAAGCA 662
Query 541 TCACTTGTGTGGCAATTCATTTGTAGTGTTCCAGTGATGGTGCTTGAAGACTTTTTGCCA 600
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Sbjct 663 TCACTTGTGTGGCAATTCATTTGTAGTGTTCCAGTGATGGTGCTTGAAGACTTTTTGCCA 722
Query 601 TGGATGATCTCTCATCTCTCTCACGAGGAGAAAATTGAAGTTGAGAATTGTATAAAAGAT 660
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Sbjct 723 TGGATGATCTCTCATCTCTCTCACGAGGAGAAAATTGAAGTTGAGAATTGTATAAAAGAT 782
Query 661 GTTGCCCCAAATGAAGACTCATTGCAACAGGTCATAAGCTCTTGGCTGCTTGACGATAGT 720
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Sbjct 783 GTTGCCCCAAATGAAGACTCATTGCAACAGGTCATAAGCTCTTGGCTGCTTGACGATAGT 842
Query 721 CAATCTTCTTGTGGGACTCCTACAGAGATCATGAAAGGAGTCCAGTATGTTAATGTGTCT 780
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Sbjct 843 CAATCTTCTTGTGGGACTCCTACAGAGATCATGAAAGGAGTCCAGTATGTTAATGTGTCT 902
Query 781 AAAAGTTTGAAGAAATCACCTGAGTCACATCCGTCAAGCGGATGTTTTCAACGCTTTTGG 840
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Sbjct 903 AAAAGTTTGAAGAAATCACCTGAGTCACATCCGTCAAGCGGATGTTTTCAACGCTTTTGG 962
Query 841 GAGTGGAGTAAAAAGTCCCTTTCAATTCCAAATGTAGGACGGAGCCCAATTCATGGTCTT 900
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Sbjct 963 GAGTGGAGTAAAAAGTCCCTTTCAATTCCAAATGTAGGACGGAGCCCAATTCATGGTCTT 1022
Query 901 AGACTTTTTCAAAATGCAATTGAAAAAGATTTGAGAGATATTCAAGAAGGATTATGCCAA 960
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Sbjct 1023 AGACTTTTTCAAAATGCAATTGAAAAAGATTTGAGAGATATTCAAGAAGGATTATGCCAA 1082
Query 961 GCAAAATTCCAAACCCTTATTTTGGATCTTGATGTACTGATGGCTAGATTAAACTTCCTT 1020
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Sbjct 1083 GCAAAATTCCAAACCCTTATTTTGGATCTTGATGTACTGATGGCTAGATTAAACTTCCTT 1142
Query 1021 GCTGATGTCCTTGTTTCTTATAGCAATGCATTTAAGAAGTTCTTTCACCCAGTGTTAGAA 1080
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Sbjct 1143 GCTGATGTCCTTGTTTCTTATAGCAATGCATTTAAGAAGTTCTTTCACCCAGTGTTAGAA 1202
Query 1081 GAAATGACAGCTCGCCGCTCTTCAACTGCCAAACAGTTCAACATAGATGATTGTCTTGAG 1140
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Sbjct 1203 GAAATGACAGCTCGCCGCTCTTCAACTGCCAAACAGTTCAACATAGATGATTGTCTTGAG 1262
Query 1141 AATTTTCAGAGATTGTTATACAAAAGTGCTGATGATAAAACAAAGACGGACAATTTTCTA 1200
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Sbjct 1263 AATTTTCAGAGATTGTTATACAAAAGTGCTGATGATAAAACAAAGACGGACAATTTTCTA 1322
Query 1201 TTGCAGCTTCAGGAGGAACTTGAGTCCCTTATAATCCAAGTAACAAAGCAATTTGCCATA 1260
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Sbjct 1323 TTGCAGCTTCAGGAGGAACTTGAGTCCCTTATAATCCAAGTAACAAAGCAATTTGCCATA 1382
Query 1261 CAGAGAACAGAGGTATTCCCAATCATCAGCAAGAATTGCAACCATGAAATGCAGAAGCAG 1320
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Sbjct 1383 CAGAGAACAGAGGTATTCCCAATCATCAGCAAGAATTGCAACCATGAAATGCAGAAGCAG 1442
Query 1321 CTCCTATACACAAGCATTCATGTCTTACCTCTTGGATTACTAAAGTGTGTCATACTTTGG 1380
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Sbjct 1443 CTCCTATACACAAGCATTCATGTCTTACCTCTTGGATTACTAAAGTGTGTCATACTTTGG 1502
Query 1381 TTTTCTGCTCATTTATCAGAAGAAGAATCTCAATCCATTCTTCATTTCTTAAGTTTGGAA 1440
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Sbjct 1503 TTTTCTGCTCATTTATCAGAAGAAGAATCTCAATCCATTCTTCATTTCTTAAGTTTGGAA 1562
Query 1441 GATTCTTCTCCCAAGAAATCATTTCCACGCCTCTTGCTACAGTGGCTTCGTTTTGGCTAT 1500
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Sbjct 1563 GATTCTTCTCCCAAGAAATCATTTCCACGCCTCTTGCTACAGTGGCTTCGTTTTGGCTAT 1622
Query 1501 TCAGGCAAAACCTCTGTTGAAAGATTTTGGAAACAGTTGGATGTTATGTTCAAAGTAAGA 1560
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Sbjct 1623 TCAGGCAAAACCTCTGTTGAAAGATTTTGGAAACAGTTGGATGTTATGTTCAAAGTAAGA 1682
Query 1561 TGTTCCTGCCAAAAGGAGCACACTGAAGAAGCATCAGGATCATTTTCCAACCAAACACAG 1620
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Sbjct 1683 TGTTCCTGCCAAAAGGAGCACACTGAAGAAGCATCAGGATCATTTTCCAACCAAACACAG 1742
Query 1621 CTGCAACTATGTAAAGTATCTAAAGATGTTTATCCTAGAAAGAAAGATAAGTCTTCAACC 1680
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Sbjct 1743 CTGCAACTATGTAAAGTATCTAAAGATGTTTATCCTAGAAAGAAAGATAAGTCTTCAACC 1802
Query 1681 TGCTTCATGTCAATGGATCTAGCTGTTGGAGACATGTATGAGACACCTTACTCAAGTCGA 1740
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Sbjct 1803 TGCTTCATGTCAATGGATCTAGCTGTTGGAGACATGTATGAGACACCTTACTCAAGTCGA 1862
Query 1741 ATGAATCAACAAATGACTTTCTCAGGAAAGCTTAAGCCTCCTCTCCATCTCCCAGAtttt 1800
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Sbjct 1863 ATGAATCAACAAATGACTTTCTCAGGAAAGCTTAAGCCTCCTCTCCATCTCCCAGATTTT 1922
Query 1801 tttGGTGAGAAGAATATGGATGATCCTATGATTATGGATGTTAAACCAATTGACCTCCTT 1860
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Sbjct 1923 TTTGGTGAGAAGAATATGGATGATCCTATGATTATGGATGTTAAACCAATTGACCTCCTT 1982
Query 1861 TTCTTCTTCCACAAGGCAATGAAGATGGATTTGGACTACCTTGTTTGTGGATCAACAAGA 1920
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Sbjct 1983 TTCTTCTTCCACAAGGCAATGAAGATGGATTTGGACTACCTTGTTTGTGGATCAACAAGA 2042
Query 1921 TTGGCAGCCGACTTTCGTTTCCTCGCGGAGTTTCAGCAACGGTTCCATATGATAAAGTTT 1980
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Sbjct 2043 TTGGCAGCCGACTTTCGTTTCCTCGCGGAGTTTCAGCAACGGTTCCATATGATAAAGTTT 2102
Query 1981 TTGTATCAGATACATTCTGATGCAGAAGATGAGATTGCATTTCCAGCATTGGAAGCCAAG 2040
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Sbjct 2103 TTGTATCAGATACATTCTGATGCAGAAGATGAGATTGCATTTCCAGCATTGGAAGCCAAG 2162
Query 2041 GGACAGCTAAAAAACATTAGTCACTCGTTTAGCATTGATCATGAGCTAGAAACTAAACAC 2100
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Sbjct 2163 GGACAGCTAAAAAACATTAGTCACTCGTTTAGCATTGATCATGAGCTAGAAACTAAACAC 2222
Query 2101 TTCGACAAAGTTTCGTTCATTTTGAATGAGATGTCAGAGCTGAACATGTTGGTTTCTACC 2160
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Sbjct 2223 TTCGACAAAGTTTCGTTCATTTTGAATGAGATGTCAGAGCTGAACATGTTGGTTTCTACC 2282
Query 2161 ATCAATACCACTGCAGCTGACCACGACCGAAAAATGAAGTATGAGCGGTTATGTCTCAGT 2220
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Sbjct 2283 ATCAATACCACTGCAGCTGACCACGACCGAAAAATGAAGTATGAGCGGTTATGTCTCAGT 2342
Query 2221 CTCCGAGAGATTTGCAAATCAATGCACAAGCTATTGTCTGAGCATATCCAACATGAAGAA 2280
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Sbjct 2343 CTCCGAGAGATTTGCAAATCAATGCACAAGCTATTGTCTGAGCATATCCAACATGAAGAA 2402
Query 2281 ACTGAGCTTTGGGGTCTATTCAGGAACTGCTTCTCTATTGAAGAACAAGAGAAAATCATA 2340
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Sbjct 2403 ACTGAGCTTTGGGGTCTATTCAGGAACTGCTTCTCTATTGAAGAACAAGAGAAAATCATA 2462
Query 2341 GGATGCATGCTCGGGAGAATAAGTGGGGAAATATTGCAGGATATGATTCCTTGGTTAATG 2400
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Sbjct 2463 GGATGCATGCTCGGGAGAATAAGTGGGGAAATATTGCAGGATATGATTCCTTGGTTAATG 2522
Query 2401 GAATCTTTAACCTCTGATGAACAGCTTGCCGCGATGTCCTTGTGGCGCCAAGCAACAAGG 2460
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Sbjct 2523 GAATCTTTAACCTCTGATGAACAGCTTGCCGCGATGTCCTTGTGGCGCCAAGCAACAAGG 2582
Query 2461 AAAACAATGTTTGTCGAATGGCTAACAGAATGGTATAACGGTCATGTTTTACAAGAGGAA 2520
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Sbjct 2583 AAAACAATGTTTGTCGAATGGCTAACAGAATGGTATAACGGTCATGTTTTACAAGAGGAA 2642
Query 2521 GCAGGAGAAGCAAACAACGATCCATTTGGGGATTCAGATCCACTAGAGATTGTCTGGAAG 2580
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Sbjct 2643 GCAGGAGAAGCAAACAACGATCCATTTGGGGATTCAGATCCACTAGAGATTGTCTGGAAG 2702
Query 2581 TATCTCTTTGAAGCATCTGCTGATGGGGAAAAAGGGAGCATGCGCAGCAGCCTTCTCAAA 2640
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Sbjct 2703 TATCTCTTTGAAGCATCTGCTGATGGGGAAAAAGGGAGCATGCGCAGCAGCCTTCTCAAA 2762
Query 2641 CTCCCGAAGACGAATTTTACAGGTATTATGAATCAGCCTCCTCCTAATTACAAGGTTGAA 2700
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Sbjct 2763 CTCCCGAAGACGAATTTTACAGGTATTATGAATCAGCCTCCTCCTAATTACAAGGTTGAA 2822
Query 2701 GTTGGCAAAAAAGAGGAAAAAGATCTAGAACGGTCAGAAAGTAAAAAGATATGCAGAGGA 2760
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Sbjct 2823 GTTGGCAAAAAAGAGGAAAAAGATCTAGAACGGTCAGAAAGTAAAAAGATATGCAGAGGA 2882
Query 2761 TCTAACCAAGAGGGGGACAAAGAACAAACTGACAAAATGTCTCAGAAGGTTAGCCAGTTT 2820
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Sbjct 2883 TCTAACCAAGAGGGGGACAAAGAACAAACTGACAAAATGTCTCAGAAGGTTAGCCAGTTT 2942
Query 2821 GGTCCATCCAAGAAATATGAACAACTGTTAACAATGAGTGAAGAAGAACTTGTGGTTGTG 2880
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Sbjct 2943 GGTCCATCCAAGAAATATGAACAACTGTTAACAATGAGTGAAGAAGAACTTGTGGTTGTG 3002
Query 2881 ATTAAGAAAATATCATGCGACTCTTCCCTGGATCCTCAGAAGAAAGACTATATCAAGCAG 2940
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Sbjct 3003 ATTAAGAAAATATCATGCGACTCTTCCCTGGATCCTCAGAAGAAAGACTATATCAAGCAG 3062
Query 2941 AACTTGTTAATGAGTCGGTGGAATATCTCACAACGTACATATAATTTGGAACCATCCTCT 3000
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Sbjct 3063 AACTTGTTAATGAGTCGGTGGAATATCTCACAACGTACATATAATTTGGAACCATCCTCT 3122
Query 3001 CTCTCGAGCAACATGGAAACAGTTCATGGTCAACATCCATCATATCGAGATCCTCACAGT 3060
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Sbjct 3123 CTCTCGAGCAACATGGAAACAGTTCATGGTCAACATCCATCATATCGAGATCCTCACAGT 3182
Query 3061 TTGATCTTTGGTTGCAATCACTACAAGAGAAATTGTAAACTTCTTGCTCCTTGTTGTGAC 3120
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Sbjct 3183 TTGATCTTTGGTTGCAATCACTACAAGAGAAATTGTAAACTTCTTGCTCCTTGTTGTGAC 3242
Query 3121 AAGCTCTTCACATGTATACGATGCCATGATGAAGAAGCTGATCACTCGGTGGATAGGAAA 3180
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Sbjct 3243 AAGCTCTTCACATGTATACGATGCCATGATGAAGAAGCTGATCACTCGGTGGATAGGAAA 3302
Query 3181 CAGATTACAAAGATGATGTGCATGAAATGCCTCTTGATTCAACCGATTGGTGCAAATTGC 3240
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Sbjct 3303 CAGATTACAAAGATGATGTGCATGAAATGCCTCTTGATTCAACCGATTGGTGCAAATTGC 3362
Query 3241 TCAAATACTTCGTGCAAGTCGTCAATGGGAAAATACTTCTGCAAAATATGCAAATTATAT 3300
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Sbjct 3363 TCAAATACTTCGTGCAAGTCGTCAATGGGAAAATACTTCTGCAAAATATGCAAATTATAT 3422
Query 3301 GACGATGAAAGGAAAATTTATCACTGTCCTTACTGCAACCTCTGTCGAGTAGGGAAAGGA 3360
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Sbjct 3423 GACGATGAAAGGAAAATTTATCACTGTCCTTACTGCAACCTCTGTCGAGTAGGGAAAGGA 3482
Query 3361 TTGGGTATTGACTACTTCCACTGCATGAAATGCAATGCTTGTATGTCACGTACCTTAGTA 3420
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Sbjct 3483 TTGGGTATTGACTACTTCCACTGCATGAAATGCAATGCTTGTATGTCACGTACCTTAGTA 3542
Query 3421 GAGCATGTTTGCAGGGAAAAGTGCTTAGAAGATAATTGTCCGATTTGCCATGAGTATATC 3480
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Sbjct 3543 GAGCATGTTTGCAGGGAAAAGTGCTTAGAAGATAATTGTCCGATTTGCCATGAGTATATC 3602
Query 3481 TTCACCTCCAGCTCCCCAGTGAAGGCTCTTCCTTGTGGTCACTTGATGCACTCCACATGC 3540
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Sbjct 3603 TTCACCTCCAGCTCCCCAGTGAAGGCTCTTCCTTGTGGTCACTTGATGCACTCCACATGC 3662
Query 3541 TTTCAGGAGTACACATGTTCACATTACACATGCCCTGTCTGCAGCAAGTCGCTTGGAGAT 3600
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Sbjct 3663 TTTCAGGAGTACACATGTTCACATTACACATGCCCTGTCTGCAGCAAGTCGCTTGGAGAT 3722
Query 3601 ATGCAGGTCTACTTTAAGATGTTAGACGCATTGCTTGCAGAAGAGAAGATGCCTGATGAA 3660
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Sbjct 3723 ATGCAGGTCTACTTTAAGATGTTAGACGCATTGCTTGCAGAAGAGAAGATGCCTGATGAA 3782
Query 3661 TACTCCAACAAAACTCAGGTGATCTTGTGTAATGATTGTGGAAGAAAAGGAAATGCTCCT 3720
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Sbjct 3783 TACTCCAACAAAACTCAGGTGATCTTGTGTAATGATTGTGGAAGAAAAGGAAATGCTCCT 3842
Query 3721 TACCATTGGCTTTACCACAAATGCACAACCTGTGGCTCCTACAACTCAAGGCTCCTTTAG 3780
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Sbjct 3843 TACCATTGGCTTTACCACAAATGCACAACCTGTGGCTCCTACAACTCAAGGCTCCTTTAG 3902
100.0% identity in 1259 residues overlap; Score: 6701.0; Gap frequency: 0.0%
Sequence1 Applicant’s SEQ ID NO:139
Sequence2 Amino Acid Sequence from Theologis et al 2022
Sequence1 1 MGGGNLHSLPPENASVSASYAVTVGNTKLSDAPVLFFVYCHKAFRAQLVELRRFATDAAE
Sequence2 1 MGGGNLHSLPPENASVSASYAVTVGNTKLSDAPVLFFVYCHKAFRAQLVELRRFATDAAE
************************************************************
Sequence1 61 ADSFSGDLAVELSRKFEFLKLVYKYHSAAEDEVIFLALDKRVKNIVSNYSLEHAGTDDLF
Sequence2 61 ADSFSGDLAVELSRKFEFLKLVYKYHSAAEDEVIFLALDKRVKNIVSNYSLEHAGTDDLF
************************************************************
Sequence1 121 TSIFHWLHVLEEEIGSRSDVLREVILCIGTIQSSICQHMLKEERQVFPLLIEKFSFREQA
Sequence2 121 TSIFHWLHVLEEEIGSRSDVLREVILCIGTIQSSICQHMLKEERQVFPLLIEKFSFREQA
************************************************************
Sequence1 181 SLVWQFICSVPVMVLEDFLPWMISHLSHEEKIEVENCIKDVAPNEDSLQQVISSWLLDDS
Sequence2 181 SLVWQFICSVPVMVLEDFLPWMISHLSHEEKIEVENCIKDVAPNEDSLQQVISSWLLDDS
************************************************************
Sequence1 241 QSSCGTPTEIMKGVQYVNVSKSLKKSPESHPSSGCFQRFWEWSKKSLSIPNVGRSPIHGL
Sequence2 241 QSSCGTPTEIMKGVQYVNVSKSLKKSPESHPSSGCFQRFWEWSKKSLSIPNVGRSPIHGL
************************************************************
Sequence1 301 RLFQNAIEKDLRDIQEGLCQAKFQTLILDLDVLMARLNFLADVLVSYSNAFKKFFHPVLE
Sequence2 301 RLFQNAIEKDLRDIQEGLCQAKFQTLILDLDVLMARLNFLADVLVSYSNAFKKFFHPVLE
************************************************************
Sequence1 361 EMTARRSSTAKQFNIDDCLENFQRLLYKSADDKTKTDNFLLQLQEELESLIIQVTKQFAI
Sequence2 361 EMTARRSSTAKQFNIDDCLENFQRLLYKSADDKTKTDNFLLQLQEELESLIIQVTKQFAI
************************************************************
Sequence1 421 QRTEVFPIISKNCNHEMQKQLLYTSIHVLPLGLLKCVILWFSAHLSEEESQSILHFLSLE
Sequence2 421 QRTEVFPIISKNCNHEMQKQLLYTSIHVLPLGLLKCVILWFSAHLSEEESQSILHFLSLE
************************************************************
Sequence1 481 DSSPKKSFPRLLLQWLRFGYSGKTSVERFWKQLDVMFKVRCSCQKEHTEEASGSFSNQTQ
Sequence2 481 DSSPKKSFPRLLLQWLRFGYSGKTSVERFWKQLDVMFKVRCSCQKEHTEEASGSFSNQTQ
************************************************************
Sequence1 541 LQLCKVSKDVYPRKKDKSSTCFMSMDLAVGDMYETPYSSRMNQQMTFSGKLKPPLHLPDF
Sequence2 541 LQLCKVSKDVYPRKKDKSSTCFMSMDLAVGDMYETPYSSRMNQQMTFSGKLKPPLHLPDF
************************************************************
Sequence1 601 FGEKNMDDPMIMDVKPIDLLFFFHKAMKMDLDYLVCGSTRLAADFRFLAEFQQRFHMIKF
Sequence2 601 FGEKNMDDPMIMDVKPIDLLFFFHKAMKMDLDYLVCGSTRLAADFRFLAEFQQRFHMIKF
************************************************************
Sequence1 661 LYQIHSDAEDEIAFPALEAKGQLKNISHSFSIDHELETKHFDKVSFILNEMSELNMLVST
Sequence2 661 LYQIHSDAEDEIAFPALEAKGQLKNISHSFSIDHELETKHFDKVSFILNEMSELNMLVST
************************************************************
Sequence1 721 INTTAADHDRKMKYERLCLSLREICKSMHKLLSEHIQHEETELWGLFRNCFSIEEQEKII
Sequence2 721 INTTAADHDRKMKYERLCLSLREICKSMHKLLSEHIQHEETELWGLFRNCFSIEEQEKII
************************************************************
Sequence1 781 GCMLGRISGEILQDMIPWLMESLTSDEQLAAMSLWRQATRKTMFVEWLTEWYNGHVLQEE
Sequence2 781 GCMLGRISGEILQDMIPWLMESLTSDEQLAAMSLWRQATRKTMFVEWLTEWYNGHVLQEE
************************************************************
Sequence1 841 AGEANNDPFGDSDPLEIVWKYLFEASADGEKGSMRSSLLKLPKTNFTGIMNQPPPNYKVE
Sequence2 841 AGEANNDPFGDSDPLEIVWKYLFEASADGEKGSMRSSLLKLPKTNFTGIMNQPPPNYKVE
************************************************************
Sequence1 901 VGKKEEKDLERSESKKICRGSNQEGDKEQTDKMSQKVSQFGPSKKYEQLLTMSEEELVVV
Sequence2 901 VGKKEEKDLERSESKKICRGSNQEGDKEQTDKMSQKVSQFGPSKKYEQLLTMSEEELVVV
************************************************************
Sequence1 961 IKKISCDSSLDPQKKDYIKQNLLMSRWNISQRTYNLEPSSLSSNMETVHGQHPSYRDPHS
Sequence2 961 IKKISCDSSLDPQKKDYIKQNLLMSRWNISQRTYNLEPSSLSSNMETVHGQHPSYRDPHS
************************************************************
Sequence1 1021 LIFGCNHYKRNCKLLAPCCDKLFTCIRCHDEEADHSVDRKQITKMMCMKCLLIQPIGANC
Sequence2 1021 LIFGCNHYKRNCKLLAPCCDKLFTCIRCHDEEADHSVDRKQITKMMCMKCLLIQPIGANC
************************************************************
Sequence1 1081 SNTSCKSSMGKYFCKICKLYDDERKIYHCPYCNLCRVGKGLGIDYFHCMKCNACMSRTLV
Sequence2 1081 SNTSCKSSMGKYFCKICKLYDDERKIYHCPYCNLCRVGKGLGIDYFHCMKCNACMSRTLV
************************************************************
Sequence1 1141 EHVCREKCLEDNCPICHEYIFTSSSPVKALPCGHLMHSTCFQEYTCSHYTCPVCSKSLGD
Sequence2 1141 EHVCREKCLEDNCPICHEYIFTSSSPVKALPCGHLMHSTCFQEYTCSHYTCPVCSKSLGD
************************************************************
Sequence1 1201 MQVYFKMLDALLAEEKMPDEYSNKTQVILCNDCGRKGNAPYHWLYHKCTTCGSYNSRLL
Sequence2 1201 MQVYFKMLDALLAEEKMPDEYSNKTQVILCNDCGRKGNAPYHWLYHKCTTCGSYNSRLL
***********************************************************
Accordingly, Rodríguez-Celma et al 2019, as evidenced by Theologis et al 2022, anticipates the claimed invention.
Even if it was determined that Rodríguez-Celma et al 2019 does not explicitly teach each of the limitations in the instant claims (i.e. plants are resistant against a plant pathogen as was a limitation of Claims 1 and 22), the claims are drawn to a method that results in resistance to pathogens. The method active step (i.e. reduction in expression or activity of BTSL) has been previously described by Rodríguez-Celma et al 2019. This indicates that the plants with reduced expression of BTSL in Rodríguez-Celma et al 2019 are also resistant to foliar bacterial plant pathogen (absent evidence to the contrary).
Specifically regarding obviousness, Applicants are reminded that prima facie obviousness is not rebutted by merely recognizing additional advantages or latent properties present but not recognized in the prior art. See MPEP § 2145. Mere recognition of latent properties in the prior art does not render nonobvious an otherwise known invention. Id.; In re Wiseman, 596 F.2d 1019, 201 USPQ 658 (CCPA 1979). Indeed, courts have held that “[t]he fact that appellant has recognized another advantage which would flow naturally from following the suggestion of the prior art cannot be the basis for patentability when the differences would otherwise be obvious.” Ex parte Obiaya, 227 USPQ 58, 60 (Bd. Pat. App. & Inter. 1985).
Claim Rejections - 35 USC § 103
In the event the determination of the status of the application as subject to AIA 35 U.S.C. 102 and 103 (or as subject to pre-AIA 35 U.S.C. 102 and 103) is incorrect, any correction of the statutory basis (i.e., changing from AIA to pre-AIA ) for the rejection will not be considered a new ground of rejection if the prior art relied upon, and the rationale supporting the rejection, would be the same under either status.
The following is a quotation of 35 U.S.C. 103 which forms the basis for all obviousness rejections set forth in this Office action:
A patent for a claimed invention may not be obtained, notwithstanding that the claimed invention is not identically disclosed as set forth in section 102, if the differences between the claimed invention and the prior art are such that the claimed invention as a whole would have been obvious before the effective filing date of the claimed invention to a person having ordinary skill in the art to which the claimed invention pertains. Patentability shall not be negated by the manner in which the invention was made.
Claims 13-14 are rejected under 35 U.S.C. 103 as being unpatentable over the teachings of Rodríguez-Celma et al 2019 and applied to Claims 1 and 11 in further view of WO 2024008752 A1 (Harrington) as evidenced by Shkryl, Yury, et al. "CRISPR/Cas9-mediated knockout of HOS1 reveals its role in the regulation of secondary metabolism in Arabidopsis thaliana." Plants 10.1 (2021): 104.
Rodríguez-Celma et al 2019 does not explicitly teach the limitations of one or more exogenous nucleic acid molecules comprise one or more guide nucleic acid molecules that can delete or mutate the BTSL gene (i.e. Claim 13) or the use of Cas proteins (Claim 14)
Harrington is directed to genetically altered plants, parts thereof or plant cells, where the plants, parts thereof or plant cells are characterized by increased levels of iron (see abstract). They describe mutating the BTSL homologue BRUTUS (BTS). Page 28 paragraphs 3 teaches CRISPR can be used to target gens for editing. Heterologous expression (i.e. introducing of Claim 14; introduces using a exogenous nucleic acid molecule of Claim 11 ) of Cas proteins and a guide nucleic acid (i.e. guide RNA (sgRNA)) molecule of Claim 13 are described on page 29 paragraph 2 to page 30 paragraph 2 and page 31 paragraph 2. Additionally, page 42 paragraph 2 example 2 teaches how guide RNAs ((and presumably Cas proteins) were used to cause nucleotide deletions of the BRUTUS homologue in wheat indicating that Harrington was in possession of a method for reducing BTSL homologue activity or expression.
It would have been prima facie obvious to combine the teachings of Rodríguez-Celma et al 2019 and Harrington as both are directed to reducing expression or activity of BTSL or BTS (i.e. homologs) which leads to an accumulation of iron. The methods for reduced activity in Rodríguez-Celma et al 2019 used T-DNA insertion knockout mutants while Harrington used CRISPR/Cas9 technology to reduce expression or activity. These two techniques are methods that can be used to generate the same outcome as evidenced by Shkryl et al 2021. Shkryl et al 2021 recited that knockout mutants generated with T-DNA insertions or CRISPR were similar in the passage “Mutant plants, carrying hos1 allele generated via CRISPR/Cas9 chemistry (hos1Cas9) and traditional T-DNA insertion mutagenesis (hos1-3) showed similar phenotypes, physiological and molecular responses to abiotic stress stimuli”. This gives one of ordinary skill in the art a reasonable expectation of success as Cas proteins and guide nucleic acid molecules can be used to generate mutants with similar phenotypes as T-DNA insertion Salk Lines. One with ordinary skill in the art would have been motivated to generate a method that utilizes exogenous nucleic acid molecules comprising guide nucleic acid molecules and introducing Cas protein for editing BTS/BTSL for engineered plants that increase iron accumulation to increase their nutritional value.
Conclusion
No claims are allowed.
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/GEORGE W MEYER/Examiner, Art Unit 1662
/BRATISLAV STANKOVIC/Supervisory Patent Examiner, Art Units 1661 & 1662
1 As evidenced by the abstract and SEQ ID NO:1 of CN 113372425 A which teaches Applicant’s SEQ ID NO:135 is already a known DNA sequence for encoding the At1g47400 (IMA1) protein.
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