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 .
Continued Examination Under 37 CFR 1.114
A request for continued examination under 37 CFR 1.114, including the fee set forth in 37 CFR 1.17(e), was filed in this application after final rejection. Since this application is eligible for continued examination under 37 CFR 1.114, and the fee set forth in 37 CFR 1.17(e) has been timely paid, the finality of the previous Office action has been withdrawn pursuant to 37 CFR 1.114. Applicant's submission filed on 11/18/2025 has been entered.
The rejection of claims 1-14 & 25-28 under 35 U.S.C. 112(b) or 35 U.S.C. 112 (pre-AIA ), second paragraph, is withdrawn in light of Applicant’s amendments.
The rejection of claims 7 & 11 under 35 U.S.C. 112(d) or pre-AIA 35 U.S.C. 112, 4th paragraph, is withdrawn in light of Applicant’s amendments.
The rejection of claims 1-14 & 25-28 under 35 U.S.C. 112(a) or 35 U.S.C. 112 (pre-AIA ), first paragraph, is withdrawn in light of Applicant’s amendments.
The rejection of claims 1-5 under 35 U.S.C. 103 as being unpatentable over Frenzel et al (2005) MPMI 18(8):771–782, in view of Uniprot record A9YWS7 LEC10_MEDTR is withdrawn in light of Applicant’s amendments.
The rejection of claims 1-3 & 6-14 under 35 U.S.C. 103 as being unpatentable over Etzler, Frenzel, and UniProt record A9YWS7 LEC10_MEDTR and further in view of Sanz Molinero et al (US 20100199379 A1), and UniProt record H2EST4_MEDTR is withdrawn in light of Applicant’s amendments.
The rejection of claims 1-3, 6-8, 10-11, 14 & 27-28 under 35 U.S.C. 103 as being unpatentable over Etzler et al US 6849777 B1, in view of Frenzel, and Uniprot record A9YWS7 LEC10_MEDTR and further in view of Sanz Molinero et al (US 20100199379 A1), and UniProt record H2EST4_MEDTR is withdrawn in light of Applicant’s amendments.
The rejection of claims 1, 2, 6-8 & 10-17 under 35 U.S.C. 103 as being unpatentable over Etzler, Frenzel, Uniprot record A9YWS7 LEC10_MEDTR, Vershinina, and Schmeisser, and further in view of Sanz Molinero and UniProt record H2EST4_MEDTR is withdrawn in light of Applicant’s amendments.
Status of Claims
Claims 1-2, 6-17, 25-28 are under examination on the merits.
Claims 18 -24 are withdrawn.
Claim Objections
Claim 16 is objected to because of the following informalities:
Claim 16 (lines 2-4) recites a closed Markush group consisting of both “S. meliloti” and “Sinorhizobium meliloti”. If these options refer to the same species, only one needs to be included in the Markush group. Additionally, the genera of “S. meliloti” (line 2) and “R. irregularis” (lines 5-6) should be written out as they are the first appearance of the genera in the claims.
Appropriate correction is required.
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 1, 2, 6-8 & 10-14 are rejected under 35 U.S.C. 103 as being unpatentable over Etzler et al US 6849777 B1 (filed 9/6/2000, granted 2/1/2005), hereafter Etzler, in view of Frenzel et al (2005) MPMI 18(8):771–782, hereafter Frenzel, and in view of Uniprot record A9YWS7 LEC10_MEDTR (first available 2/5/2008).
Due to Applicant' s amendment of the claims, the rejection is modified from the rejection as set forth in the Office action mailed 5/20/2025, as applied to claims 1-8 & 10-14. Applicant' s arguments filed 11/18/2025 have been fully considered but they are not persuasive.
Claims 1, 2, 6-8 & 10-14 are drawn to a transgenic plant cell comprising a DNA molecule encoding a lectin-domain containing protein comprising the amino acid sequence of SEQ ID NO: 2 and a method of making progeny seed.
Etzler teaches a method of increasing mycorrhizal infection in a plant comprising introducing polynucleotide for a LNP protein (Etzler claim 1, lines 1-8). Etzler teaches that the mycorrhizal fungus symbiont can be Glomus intraradices (Etzler claim 6, lines 1-2; and column 20 lines 18-38), although Etzler also teaches a method of associating transgenic plants with Mesorhizobium loti bacteria (column 17, lines 1-6 table 1; column 19, line 31-column 20, line 16).
Etzler teaches that LNP is a root lectin protein (column 1, line 66) involved in rhizobium-legume biosynthesis (column 2, lines 5-6) and nodulation (table 1 and column 18, lines 7-17). Etzler teaches that LNP is able to bind Nod factors (column 16, line 43). Etzler teaches suitable heterologous promoters, including the CaMV 35S promoter and teaches a motivation for using a heterologous promoter to achieve constitutive and ubiquitous expression (column 13, lines 48-51).
Etzler teaches stably transformed lines of L. japonicus, a dicot, via Agrobacterium transformation (column 19, lines 6-30) and teaches planting and growing seeds from these lines specifically to the T1 and T2 generation (column 19, lines 20-37 and table 1). Etzler teaches a variant of the method of increasing mycorrhizal infection where the cassette is introduced into the plant through a sexual cross (Etzler claim 4).
Etzler does not teach instant SEQ ID NO: 2. Etzler does not teach the transformation of M. truncatula. Etzler does not specifically teach a method of producing progeny seed comprising plant a first seed, growing a plant from the seed, and harvesting the progeny seed from the plants.
Frenzel teaches a cDNA library created from mycorrhizal roots of Medicago truncatula and enriched for mycorrhiza-upregulated genes, ligated into the pGEM-T vector in E. coli (page 779, right column, lines 19-31). Frenzel teaches that lectin genes are strongly induced in mycorrhizal roots, including the MtLEC10 gene (page 776, left column, lines 6-17). Furthermore, Frenzel teaches that MtLEC10 (TC 75254) is detected only in mycorrhizal roots and not in non-mycorrhizal roots (page 775, table 4). Frenzel suggests hypothetical functions for the lectin proteins, including storage for organic nitrogen, and suggests that AM-specific lectins might have different functions than nodule-enhanced lectins (page 779, left column, paragraph 1).
Uniprot record A9YWS7 LEC10_MEDTR teaches a sequence identical to instant SEQ ID NO: 2. See alignment below.
LEC10_MEDTR
ID LEC10_MEDTR Reviewed; 287 AA.
AC A9YWS7; A0A0C3XFZ3; G7KGH3;
DT 17-JUN-2020, integrated into UniProtKB/Swiss-Prot.
DT 05-FEB-2008, sequence version 1.
DT 24-JAN-2024, entry version 75.
DE RecName: Full=Lectin 10 {ECO:0000303|PubMed:16134889};
DE Short=MtLec10 {ECO:0000303|PubMed:16134889};
DE AltName: Full=Agglutinin LEC10 {ECO:0000305};
GN Name=LEC10 {ECO:0000303|PubMed:16134889};
GN OrderedLocusNames=MTR_5g031160 {ECO:0000312|EMBL:AES95928.2};
GN ORFNames=MtrunA17_Chr5g0411771 {ECO:0000312|EMBL:RHN54888.1};
OS Medicago truncatula (Barrel medic) (Medicago tribuloides).
OC Eukaryota; Viridiplantae; Streptophyta; Embryophyta; Tracheophyta;
OC Spermatophyta; Magnoliopsida; eudicotyledons; Gunneridae; Pentapetalae;
OC rosids; fabids; Fabales; Fabaceae; Papilionoideae; 50 kb inversion clade;
OC NPAAA clade; Hologalegina; IRL clade; Trifolieae; Medicago.
OX NCBI_TaxID=3880;
RN [1]
RP NUCLEOTIDE SEQUENCE [LARGE SCALE GENOMIC DNA].
RC STRAIN=cv. Jemalong A17;
RX PubMed=16273388; DOI=10.1007/s00438-005-0057-9;
RA Kevei Z., Seres A., Kereszt A., Kalo P., Kiss P., Toth G., Endre G.,
RA Kiss G.B.;
RT "Significant microsynteny with new evolutionary highlights is detected
RT between Arabidopsis and legume model plants despite the lack of
RT macrosynteny.";
RL Mol. Genet. Genomics 274:644-657(2005).
RN [2]
RP NUCLEOTIDE SEQUENCE [LARGE SCALE GENOMIC DNA].
RC STRAIN=cv. Jemalong A17;
RX PubMed=22089132; DOI=10.1038/nature10625;
RA Young N.D., Debelle F., Oldroyd G.E.D., Geurts R., Cannon S.B.,
RA Udvardi M.K., Benedito V.A., Mayer K.F.X., Gouzy J., Schoof H.,
RA Van de Peer Y., Proost S., Cook D.R., Meyers B.C., Spannagl M., Cheung F.,
RA De Mita S., Krishnakumar V., Gundlach H., Zhou S., Mudge J., Bharti A.K.,
RA Murray J.D., Naoumkina M.A., Rosen B., Silverstein K.A.T., Tang H.,
RA Rombauts S., Zhao P.X., Zhou P., Barbe V., Bardou P., Bechner M.,
RA Bellec A., Berger A., Berges H., Bidwell S., Bisseling T., Choisne N.,
RA Couloux A., Denny R., Deshpande S., Dai X., Doyle J.J., Dudez A.-M.,
RA Farmer A.D., Fouteau S., Franken C., Gibelin C., Gish J., Goldstein S.,
RA Gonzalez A.J., Green P.J., Hallab A., Hartog M., Hua A., Humphray S.J.,
RA Jeong D.-H., Jing Y., Jocker A., Kenton S.M., Kim D.-J., Klee K., Lai H.,
RA Lang C., Lin S., Macmil S.L., Magdelenat G., Matthews L., McCorrison J.,
RA Monaghan E.L., Mun J.-H., Najar F.Z., Nicholson C., Noirot C.,
RA O'Bleness M., Paule C.R., Poulain J., Prion F., Qin B., Qu C., Retzel E.F.,
RA Riddle C., Sallet E., Samain S., Samson N., Sanders I., Saurat O.,
RA Scarpelli C., Schiex T., Segurens B., Severin A.J., Sherrier D.J., Shi R.,
RA Sims S., Singer S.R., Sinharoy S., Sterck L., Viollet A., Wang B.-B.,
RA Wang K., Wang M., Wang X., Warfsmann J., Weissenbach J., White D.D.,
RA White J.D., Wiley G.B., Wincker P., Xing Y., Yang L., Yao Z., Ying F.,
RA Zhai J., Zhou L., Zuber A., Denarie J., Dixon R.A., May G.D.,
RA Schwartz D.C., Rogers J., Quetier F., Town C.D., Roe B.A.;
RT "The Medicago genome provides insight into the evolution of rhizobial
RT symbioses.";
RL Nature 480:520-524(2011).
RN [3]
RP GENOME REANNOTATION.
RC STRAIN=cv. Jemalong A17;
RX PubMed=24767513; DOI=10.1186/1471-2164-15-312;
RA Tang H., Krishnakumar V., Bidwell S., Rosen B., Chan A., Zhou S.,
RA Gentzbittel L., Childs K.L., Yandell M., Gundlach H., Mayer K.F.,
RA Schwartz D.C., Town C.D.;
RT "An improved genome release (version Mt4.0) for the model legume Medicago
RT truncatula.";
RL BMC Genomics 15:312-312(2014).
RN [4]
RP NUCLEOTIDE SEQUENCE [LARGE SCALE GENOMIC DNA].
RC STRAIN=cv. Jemalong A17;
RX PubMed=30397259; DOI=10.1038/s41477-018-0286-7;
RA Pecrix Y., Staton S.E., Sallet E., Lelandais-Briere C., Moreau S.,
RA Carrere S., Blein T., Jardinaud M.F., Latrasse D., Zouine M., Zahm M.,
RA Kreplak J., Mayjonade B., Satge C., Perez M., Cauet S., Marande W.,
RA Chantry-Darmon C., Lopez-Roques C., Bouchez O., Berard A., Debelle F.,
RA Munos S., Bendahmane A., Berges H., Niebel A., Buitink J., Frugier F.,
RA Benhamed M., Crespi M., Gouzy J., Gamas P.;
RT "Whole-genome landscape of Medicago truncatula symbiotic genes.";
RL Nat. Plants 4:1017-1025(2018).
RN [5]
RP INDUCTION BY ARBUSCULAR MYCORRHIZAL FUNGI.
RC STRAIN=cv. Jemalong A17;
RX PubMed=16134889; DOI=10.1094/mpmi-18-0771;
RA Frenzel A., Manthey K., Perlick A.M., Meyer F., Puehler A., Kuester H.,
RA Krajinski F.;
RT "Combined transcriptome profiling reveals a novel family of arbuscular
RT mycorrhizal-specific Medicago truncatula lectin genes.";
RL Mol. Plant Microbe Interact. 18:771-782(2005).
CC -!- FUNCTION: May be involved in arbuscular mycorrhizal (AM) symbiosis with
CC AM fungi. {ECO:0000305|PubMed:16134889}.
CC -!- SUBCELLULAR LOCATION: Membrane {ECO:0000255}; Single-pass membrane
CC protein {ECO:0000255}.
CC -!- INDUCTION: Accumulates in roots during colonization by arbuscular
CC mycorrhizal (AM) fungi (e.g. Glomus intraradices).
CC {ECO:0000269|PubMed:16134889}.
CC -!- SIMILARITY: Belongs to the leguminous lectin family. {ECO:0000305}.
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DR EMBL; EU306659; ABY48148.1; -; Genomic_DNA.
DR EMBL; CM001221; AES95928.2; -; Genomic_DNA.
DR EMBL; PSQE01000005; RHN54888.1; -; Genomic_DNA.
DR RefSeq; XP_003612970.2; XM_003612922.2.
DR AlphaFoldDB; A9YWS7; -.
DR SMR; A9YWS7; -.
DR STRING; 3880.A9YWS7; -.
DR GlyCosmos; A9YWS7; 4 sites, No reported glycans.
DR PaxDb; 3880-AES95928; -.
DR EnsemblPlants; AES95928; AES95928; MTR_5g031160.
DR GeneID; 11405785; -.
DR Gramene; AES95928; AES95928; MTR_5g031160.
DR KEGG; mtr:11405785; -.
DR eggNOG; ENOG502QTX3; Eukaryota.
DR HOGENOM; CLU_000288_62_2_1; -.
DR OrthoDB; 513960at2759; -.
DR Proteomes; UP000002051; Chromosome 5.
DR Proteomes; UP000265566; Chromosome 5.
DR GO; GO:0016020; C:membrane; IEA:UniProtKB-SubCell.
DR GO; GO:0030246; F:carbohydrate binding; IEA:UniProtKB-KW.
DR GO; GO:0009610; P:response to symbiotic fungus; IEP:UniProtKB.
DR CDD; cd06899; lectin_legume_LecRK_Arcelin_ConA; 1.
DR Gene3D; 2.60.120.200; -; 1.
DR InterPro; IPR013320; ConA-like_dom_sf.
DR InterPro; IPR016363; L-lectin.
DR InterPro; IPR000985; Lectin_LegA_CS.
DR InterPro; IPR001220; Legume_lectin_dom.
DR PANTHER; PTHR32401; CONCANAVALIN A-LIKE LECTIN FAMILY PROTEIN; 1.
DR PANTHER; PTHR32401:SF31; LECTIN 6; 1.
DR Pfam; PF00139; Lectin_legB; 1.
DR PIRSF; PIRSF002690; L-type_lectin_plant; 1.
DR SUPFAM; SSF49899; Concanavalin A-like lectins/glucanases; 1.
DR PROSITE; PS00308; LECTIN_LEGUME_ALPHA; 1.
PE 2: Evidence at transcript level;
KW Glycoprotein; Lectin; Membrane; Reference proteome; Transmembrane;
KW Transmembrane helix.
FT CHAIN 1..287
FT /note="Lectin 10"
FT /id="PRO_0000450045"
FT TOPO_DOM 1..11
FT /note="Cytoplasmic"
FT /evidence="ECO:0000305"
FT TRANSMEM 12..31
FT /note="Helical"
FT /evidence="ECO:0000255"
FT TOPO_DOM 32..287
FT /note="Extracellular"
FT /evidence="ECO:0000305"
FT CARBOHYD 124
FT /note="N-linked (GlcNAc...) asparagine"
FT /evidence="ECO:0000255|PROSITE-ProRule:PRU00498"
FT CARBOHYD 147
FT /note="N-linked (GlcNAc...) asparagine"
FT /evidence="ECO:0000255|PROSITE-ProRule:PRU00498"
FT CARBOHYD 243
FT /note="N-linked (GlcNAc...) asparagine"
FT /evidence="ECO:0000255|PROSITE-ProRule:PRU00498"
FT CARBOHYD 280
FT /note="N-linked (GlcNAc...) asparagine"
FT /evidence="ECO:0000255|PROSITE-ProRule:PRU00498"
SQ SEQUENCE 287 AA; 31923 MW; 8D96C83144D88E97 CRC64;
Query Match 100.0%; Score 1502; Length 287;
Best Local Similarity 100.0%;
Matches 287; Conservative 0; Mismatches 0; Indels 0; Gaps 0;
Qy 1 MALSNLKSNRTLSSSLITIFIISLFLQYHNIKSQSSWQSRQVPRSETVAFSITEFEKENP 60
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 1 MALSNLKSNRTLSSSLITIFIISLFLQYHNIKSQSSWQSRQVPRSETVAFSITEFEKENP 60
Qy 61 DIFLRGDTSISDGILRLTKTDQSGKPLPNTVGRATYLTPIHIWDKTSGELADFSTSFSFI 120
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 61 DIFLRGDTSISDGILRLTKTDQSGKPLPNTVGRATYLTPIHIWDKTSGELADFSTSFSFI 120
Qy 121 VNTNDSDLHGDGFAFYLGPLHFDVPKNSSGGYLGLFDPENAFPPSKTPILAIEFDGFTNE 180
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 121 VNTNDSDLHGDGFAFYLGPLHFDVPKNSSGGYLGLFDPENAFPPSKTPILAIEFDGFTNE 180
Qy 181 WDPPSSFQSPHIGIDVGSIVSLEYAQWPINFVPRNALGEANINYNSESKRLSVFVAYPGT 240
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 181 WDPPSSFQSPHIGIDVGSIVSLEYAQWPINFVPRNALGEANINYNSESKRLSVFVAYPGT 240
Qy 241 QWNSTRVSVVVDLRSVLPEWVRIGFSATTGELVETHDIINWSFESAL 287
|||||||||||||||||||||||||||||||||||||||||||||||
Db 241 QWNSTRVSVVVDLRSVLPEWVRIGFSATTGELVETHDIINWSFESAL 287
Before the time of filing of the instant application, it would have been obvious to one having ordinary skill in the art to substitute the lectin-domain containing protein taught by Uniprot record A9YWS7 LEC10_MEDTR for the lectin protein used by Etzler to increase mycorrhizal infection in a plant. One of ordinary skill in the art would have been motivated to make the substitution because MtLEC10 was taught to be a lectin protein involved in mycorrhizal infection. One of ordinary skill in the art would have been reasonably confident of success in substituting the MtLEC10 gene, because it was expressed in roots and believed to be involved in mycorrhizal infection in a legume plant. Furthermore, it would have been obvious to one of ordinary skill that a MtLEC10 gene taught by Frenzel would encode a protein with a Lec10 sequence of A9YWS7 LEC10_MEDTR, which has 100% homology to instant SEQ ID NO: 2.
Regarding claims 1-2, Etzler teaches a lectin protein-encoding polynucleotide operably linked to a plant promoter or a CaMV35S promoter in an expression cassette in a plant to increase mycorrhizal infection. The expression cassette reads on a heterologous promoter operably linked to the polynucleotide segment. The recombinant DNA molecule encoding a lectin-domain protein taught by A9YWS7 LEC10_MEDTR, because it has 100% sequence homology to instant SEQ ID NO: 2, would have been capable of producing membrane invagination when expressed in a plant cell. Furthermore, Etzler teaches that the method of introducing an expression cassette containing the plant promoter linked to the heterologous LNP polynucleotide is a method to increase mycorrhizal infection, which reads on a plant cell exhibiting an increase in symbiotic infection of a fungus (instant claim 2).
Etzler teaches introducing an expression cassette containing a plant promoter linked to a heterologous LNP polynucleotide into a plant to increase mycorrhizal infection (Etzler claim 1), and specifically teaches this in Lotus japonicus. Increased mycorrhizal infection reads on a plant susceptible to symbiotic infection (instant claim 14, line 1). Thus, instant claims 6, 8, 10, and 14 to a plant or plant cell comprising the recombinant DNA molecule of instant claim 1 and susceptible to symbiotic infection are obvious.
Regarding claims 7 and 11, before the time of filing of the instant application, it would have been obvious to express a recombinant DNA molecule comprising a gene with the sequence of MtLEC10 in M. truncatula, because MtLEC10 originates in M. truncatula.
Regarding claims 12 and 13, Etzler taught generating stably transformed lines for producing seed for experiment, including planting a seed and growing a plant from the seed until the T2 generation. Therefore, before the time of filing of the instant application, it would have been obvious to one having ordinary skill in the art to produce progeny seed from a transgenic plant comprising the recombinant DNA by planting a transgenic seed, growing a plant from the seed, and harvesting the progeny seed from the plants. The step of harvesting the progeny seed from the plants would be inherent in order to acquire seed for using in the experiments. Thus, the method of making progeny seed is obvious in light of the seed taught by Etzler.
Claims 1-2, 6-8 & 10-14 are obvious in view of Etzler, Frenzel, and Uniprot record A9YWS7 LEC10_MEDTR.
Applicant urges that the LNP sequence of Etzler does not structurally resemble instant SEQ ID NO: 2 with 8% sequence identity and no regions of short overlap. Applicant urges that Etzler’s LNP is not a conventional legume but a lectin-nucleotide phosphohydrolase with dual biochemical activity. Applicant urges that Etzler attributes the symbiosis phenotype to apyrase catalysis and a role upstream and in a different location compared to infection thread stage. Applicant urges that because of the 1) structure, 2) biochemical activity and proposed mechanism of LNP, and 3) spatial and temporal expression, one of skill in the art would not be motivated to select a sequence of LEC10 with 100% homology to instant SEQ ID NO: 2 because they would not have been regarded as functional equivalents (Remarks, page 15, paragraph 2-page 16 paragraph 3).
This argument is unpersuasive, because although the specific role of the individual protein with 100% sequence identity to SEQ ID NO: 2 was not known, Franzel teaches that the accumulation of transcripts of lectin genes, including Lec10, indicates an important role for these proteins during AM symbiosis, (page 779, left column, paragraph 1-right column, paragraph 1).
1) Lectin proteins are known to differ greatly, and some lectins are known to function as determinants of specificity (see eg Franzel, page 779, left column, paragraph 1), so low sequence identity between Etzler SEQ ID NO: 10 and Uniprot record A9YWS7 LEC10_MEDTR would not in itself be evidence that the proteins taught by Franzel to be involved in and important to arbuscular mycorrhizal symbiosis would not also promote arbuscular mycorrhizal symbiosis.
2) Etzler teaches apyrase activity of LNP in the context of enhancement of phosphate uptake after infection by mycorrhizal fungi (column 9, lines 33-40). Etzler also teaches that the LNP protein modulates oligosaccharide signaling events important for the interaction between mycorrhizal fungi and plants (column 9, lines 33-40). Etzler does not teach that the apyrase activity is required for promotion of mycorrhizal association, so one of ordinary skill in the art would not have concluded, based on Etzler, that the proteins taught by Franzel to be involved in arbuscular mycorrhizal symbiosis would not also promote arbuscular mycorrhizal symbiosis.
3) Franzel does not teach the temporal expression of MtLEC10 in relation to infection threads, because Franzel teaches MtLEC10 is specific to AM symbiosis. Furthermore, Franzel is silent as to the precise timing or tissue of the expression, because at least one of the cDNA libraries was created from roots comprising all developmental stages of Glomus intraradices colonization (Franzel, page 772, left column, paragraph 3-right column, paragraph 1). Franzel teaches an important role for MtLEC10 in mycorrhizal symbiosis based on the expression profile available, so one of skill in the art would have concluded that MtLEC10 is important to mycorrhizal symbiosis without knowledge of precise spatial or temporal expression in comparison to Etzler’s LNP.
Applicant urges that one of ordinary skill in the art would not have had reasonable expectation of success because the LNP protein of Etzler does not resemble UniProt Accession A9YWS7 and belongs to an apyrase enzyme class with domains and activity that is absent from the protein of the instant claims. Applicant urges that there is no significant identity between Etzler’s LNPs and MtLEC10 beyond superficial lectin motifs. Applicant urges that because Etzler’s LNPs require divalent metal ions and act on nucleotides, and MtLEC10 does not, the lectin proteins are functionally incompatible and have different cellular contexts (Remarks page 16 paragraph 4- page 18 paragraph 3).
This argument is unpersuasive, because the rationale for MtLEC10 being important to mycorrhizal symbiotic infection was not based on mechanism or functional activity. Franzel does not teach an explicit function for the protein comprising a sequence of 100% identity to instant SEQ ID NO: 2. However, such a protein (MtLEC10) was suggested in the art to be important to mycorrhizal symbiosis prior to the instant filing, even without knowledge of the mechanism of membrane invagination at infection interfaces. The rationale to arrive at a claimed invention does not need to be Applicant’s own, so knowledge of the mechanism of membrane invagination was not necessary for one of skill in the art to have known or reasonably expected that a protein with 100% sequence identity to SEQ ID NO: 2 would be important to mycorrhizal symbiotic infection.
Furthermore, Etzler does not teach apyrase activity to be necessary for the increase in mycorrhizal infection, as explained above. The instant claims do not require that the lectin-domain containing protein lack apyrase motifs of functionality. One of skill in the art would have been motivated to express a lectin-domain containing protein such as MtLEC10 in Etzler’s method of increasing mycorrhizal infection because it was a lectin protein believed to be important to mycorrhizal symbiosis based on expression.
Applicant urges that due to structural differences from other lectin proteins, including the lack of an intracellular kinase domain, a person of skill in the art would not have expected LDP1 to be directly involved in membrane invagination or the initiation of signaling cascades. So, a person of ordinary skill in the art would not have had any teaching or motivation to select the sequence of UniProt A9YWS7 from among numerous plant lectins. Applicant urges that the present inventors have shown a surprising function of DNA molecules encoding SEQ ID NO: 2 to induce membrane invagination which is observed at a different location than the activity of LNP. Applicant urges that these surprising effects are recited in functional limitations of the claims (Remarks, page 18, paragraph 4-page 19, paragraph 3).
This argument is unpersuasive, because Franzel teaches, based on expression patterns, that a protein comprising a sequence with 100% identity to SEQ ID NO: 2 is important to mycorrhizal infection. Although there are numerous plant lectins, Franzel teaches only 7 mycorrhizal-induced lectins, including MtLEC10. The rationale to arrive at a claimed invention does not need to be Applicant’s own, so knowledge of the mechanism of membrane invagination was not necessary for one of skill in the art to have known or reasonably expected that a protein with 100% sequence identity to SEQ ID NO: 2 would be important to mycorrhizal symbiotic infection.
A DNA molecule encoding a lectin-domain protein of A9YWS7 LEC10_MEDTR, because it has 100% sequence homology to instant SEQ ID NO: 2, would have been capable of producing membrane invagination when expressed in a plant cell, even if this functionality would not have been a motivation for one of skill in the art to introduce the sequence into a plant cell.
Claims 9 & 27-28 are rejected under 35 U.S.C. 103 as being unpatentable over Etzler, Frenzel, and UniProt record A9YWS7 LEC10_MEDTR as applied to claims 1-2, 6-8, and 10-14 above, and further in view of Sanz Molinero et al (US 20100199379 A1, published 8/5/2010), hereafter Sanz Molinero; and UniProt record H2EST4_MEDTR (first available 3/21/2012).
This is a new rejection from the rejection set forth in the Office action mailed 5/20/2025, as applied to claims 1-3,6-9 & 10-14 or as applied to claims 1-3, 6-8, 10-11, 14 & 27-28. Applicant' s arguments filed 11/18/2025 have been fully considered as they apply to this rejection, but they are not persuasive.
Claim 9 is drawn to the plant of claim 8 further comprising a recombinant DNA molecule with a heterologous promoter linked to a polynucleotide encoding a SYMREM1 protein of SEQ ID NO: 14. Claim 27 is drawn to the plant of claim 8 further comprising a promoter linked to a nucleotide encoding a remorin protein with the sequence of SEQ ID NO: 25. Claim 28 is drawn to the plant of claim 27 further comprising a promoter linked to a nucleotide encoding a SYMREM1 protein with the sequence of SEQ ID NO: 14.
The teachings of Etzler, Frenzel, and Uniprot record A9YWS7 LEC10_MEDTR are summarized above. They do not teach a plant comprising a recombinant DNA molecule encoding a protein with the amino acid sequence of SEQ ID NO: 25 or 14.
Sanz Molinero teaches a rice plant comprising a recombinant expression vector encoding a remorin protein from Arabidopsis (paragraph [0836], paragraph [0839]) operably linked to the heterologous promoter pGOS2 (paragraph [0838]). Sanz Molinero teaches At4g00670 as a locus encoding a protein with homology to a remorin (page 80, table Q). At4g00670 has 100% sequence identity with SEQ ID NO: 25, see alignment below. Sanz Molinero considers the transformation of legume plants such as soybean (page 90, paragraph [0858]) and alfalfa (page 90, paragraph [0860]). Sanz Molinero teaches that the introduction of remorin protein into a plant provides a significant increase in yield (page 89, paragraph [0851] & table V).
A0A178UTR4_ARATH
(NOTE: this sequence has 3 duplicates in the database searched.
See complete list at the end of this report)
ID A0A178UTR4_ARATH Unreviewed; 123 AA.
AC A0A178UTR4;
DT 12-APR-2017, integrated into UniProtKB/TrEMBL.
DT 12-APR-2017, sequence version 1.
DT 27-MAR-2024, entry version 30.
DE SubName: Full=(thale cress) hypothetical protein {ECO:0000313|EMBL:CAD5326726.1};
GN OrderedLocusNames=At4g00670 {ECO:0000313|Araport:AT4G00670},
GN AXX17_At4g00830 {ECO:0000313|EMBL:OAO97003.1};
GN ORFNames=AN1_LOCUS16836 {ECO:0000313|EMBL:VYS61404.1},
GN AT9943_LOCUS14472 {ECO:0000313|EMBL:CAD5326726.1}, C24_LOCUS16709
GN {ECO:0000313|EMBL:CAA0392831.1};
OS Arabidopsis thaliana (Mouse-ear cress).
OC Eukaryota; Viridiplantae; Streptophyta; Embryophyta; Tracheophyta;
OC Spermatophyta; Magnoliopsida; eudicotyledons; Gunneridae; Pentapetalae;
OC rosids; malvids; Brassicales; Brassicaceae; Camelineae; Arabidopsis.
OX NCBI_TaxID=3702 {ECO:0000313|EMBL:OAO97003.1, ECO:0000313|Proteomes:UP000078284};
RN [1] {ECO:0000313|Proteomes:UP000078284}
RP NUCLEOTIDE SEQUENCE [LARGE SCALE GENOMIC DNA].
RC STRAIN=cv. Landsberg erecta {ECO:0000313|Proteomes:UP000078284};
RX PubMed=27354520; DOI=10.1073/pnas.1607532113;
RA Zapata L., Ding J., Willing E.M., Hartwig B., Bezdan D., Jiao W.B.,
RA Patel V., Velikkakam James G., Koornneef M., Ossowski S., Schneeberger K.;
RT "Chromosome-level assembly of Arabidopsis thaliana Ler reveals the extent
RT of translocation and inversion polymorphisms.";
RL Proc. Natl. Acad. Sci. U.S.A. 113:E4052-E4060(2016).
RN [2] {ECO:0000313|EMBL:OAO97003.1}
RP NUCLEOTIDE SEQUENCE [LARGE SCALE GENOMIC DNA].
RC TISSUE=Leaf {ECO:0000313|EMBL:OAO97003.1};
RA Zapata L., Schneeberger K., Ossowski S.;
RT "Full-length assembly of Arabidopsis thaliana Ler reveals the complement of
RT translocations and inversions.";
RL Submitted (MAR-2016) to the EMBL/GenBank/DDBJ databases.
RN [3] {ECO:0000313|EMBL:VYS61404.1, ECO:0000313|Proteomes:UP000426265}
RP NUCLEOTIDE SEQUENCE [LARGE SCALE GENOMIC DNA].
RC STRAIN=cv. An-1 {ECO:0000313|Proteomes:UP000426265}, and cv. C24
RC {ECO:0000313|Proteomes:UP000434276};
RA Jiao W.-B., Schneeberger K.;
RL Submitted (NOV-2019) to the EMBL/GenBank/DDBJ databases.
RN [4] {ECO:0000313|EMBL:CAD5326726.1, ECO:0000313|Proteomes:UP000516314}
RP NUCLEOTIDE SEQUENCE [LARGE SCALE GENOMIC DNA].
RC STRAIN=cv. Cdm-0 {ECO:0000313|Proteomes:UP000516314};
RA Ashkenazy H.;
RL Submitted (SEP-2020) to the EMBL/GenBank/DDBJ databases.
CC -!- SIMILARITY: Belongs to the remorin family.
CC {ECO:0000256|ARBA:ARBA00005711}.
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DR EMBL; CACSHJ010000095; CAA0392831.1; -; Genomic_DNA.
DR EMBL; LR881469; CAD5326726.1; -; Genomic_DNA.
DR EMBL; LUHQ01000004; OAO97003.1; -; Genomic_DNA.
DR EMBL; CACRSJ010000109; VYS61404.1; -; Genomic_DNA.
DR RefSeq; NP_191976.2; NM_116292.5.
DR AlphaFoldDB; A0A178UTR4; -.
DR SMR; A0A178UTR4; -.
DR EnsemblPlants; AT4G00670.1; AT4G00670.1; AT4G00670.
DR GeneID; 828034; -.
DR Gramene; AT4G00670.1; AT4G00670.1; AT4G00670.
DR KEGG; ath:AT4G00670; -.
DR Araport; AT4G00670; -.
DR OMA; MEPNIPI; -.
DR OrthoDB; 666774at2759; -.
DR Proteomes; UP000078284; Chromosome 4.
DR Proteomes; UP000426265; Unassembled WGS sequence.
DR Proteomes; UP000434276; Unassembled WGS sequence.
DR Proteomes; UP000516314; Chromosome 4.
DR ExpressionAtlas; A0A178UTR4; baseline and differential.
DR GO; GO:1901700; P:response to oxygen-containing compound; IEA:UniProt.
DR InterPro; IPR005516; Remorin_C.
DR PANTHER; PTHR31775; OS02G0117200 PROTEIN; 1.
DR PANTHER; PTHR31775:SF25; REMORIN C-TERMINAL DOMAIN-CONTAINING PROTEIN; 1.
DR Pfam; PF03763; Remorin_C; 1.
PE 3: Inferred from homology;
FT DOMAIN 12..117
FT /note="Remorin C-terminal"
FT /evidence="ECO:0000259|Pfam:PF03763"
SQ SEQUENCE 123 AA; 14089 MW; 66C9B43CF95A8CFA CRC64;
Query Match 100.0%; Score 612; Length 123;
Best Local Similarity 100.0%;
Matches 123; Conservative 0; Mismatches 0; Indels 0; Gaps 0;
Qy 1 MEPNIPIQRGDEQSKVIKAWKELKITKVNNKTQKKLLDISGWEKKKTTKIESELARIQRK 60
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 1 MEPNIPIQRGDEQSKVIKAWKELKITKVNNKTQKKLLDISGWEKKKTTKIESELARIQRK 60
Qy 61 MDSKKMEKSEKLRNEKAAVHAKAQKKKADVQTRRAQEILDAEEAAARFQAAGKIPKKSSL 120
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 61 MDSKKMEKSEKLRNEKAAVHAKAQKKKADVQTRRAQEILDAEEAAARFQAAGKIPKKSSL 120
Qy 121 SCF 123
|||
Db 121 SCF 123
UniProt record H2EST4_MEDTR teaches a SYMREM1 remorin gene with 100% sequence identity to SEQ ID NO: 14. See alignment below.
H2EST4_MEDTR
ID H2EST4_MEDTR Unreviewed; 205 AA.
AC H2EST4;
DT 21-MAR-2012, integrated into UniProtKB/TrEMBL.
DT 21-MAR-2012, sequence version 1.
DT 24-JAN-2024, entry version 45.
DE SubName: Full=Carboxy-terminal region remorin {ECO:0000313|EMBL:KEH21042.1};
DE SubName: Full=Putative remorin {ECO:0000313|EMBL:RHN43320.1};
DE SubName: Full=Symbiotic remorin 1 {ECO:0000313|EMBL:AEX20500.1};
GN Name=SYMREM1 {ECO:0000313|EMBL:AEX20500.1};
GN Synonyms=25502038 {ECO:0000313|EnsemblPlants:KEH21042};
GN OrderedLocusNames=MTR_8g097320 {ECO:0000313|EMBL:KEH21042.1};
GN ORFNames=MtrunA17_Chr8g0386521 {ECO:0000313|EMBL:RHN43320.1};
OS Medicago truncatula (Barrel medic) (Medicago tribuloides).
OC Eukaryota; Viridiplantae; Streptophyta; Embryophyta; Tracheophyta;
OC Spermatophyta; Magnoliopsida; eudicotyledons; Gunneridae; Pentapetalae;
OC rosids; fabids; Fabales; Fabaceae; Papilionoideae; 50 kb inversion clade;
OC NPAAA clade; Hologalegina; IRL clade; Trifolieae; Medicago.
OX NCBI_TaxID=3880 {ECO:0000313|EMBL:AEX20500.1};
RN [1] {ECO:0000313|EMBL:KEH21042.1, ECO:0000313|Proteomes:UP000002051}
RP NUCLEOTIDE SEQUENCE [LARGE SCALE GENOMIC DNA].
RC STRAIN=A17 {ECO:0000313|EMBL:KEH21042.1}, and cv. Jemalong A17
RC {ECO:0000313|EnsemblPlants:KEH21042,
RC ECO:0000313|Proteomes:UP000002051};
RX PubMed=22089132; DOI=10.1038/nature10625;
RA Young N.D., Debelle F., Oldroyd G.E., Geurts R., Cannon S.B., Udvardi M.K.,
RA Benedito V.A., Mayer K.F., Gouzy J., Schoof H., Van de Peer Y., Proost S.,
RA Cook D.R., Meyers B.C., Spannagl M., Cheung F., De Mita S.,
RA Krishnakumar V., Gundlach H., Zhou S., Mudge J., Bharti A.K., Murray J.D.,
RA Naoumkina M.A., Rosen B., Silverstein K.A., Tang H., Rombauts S.,
RA Zhao P.X., Zhou P., Barbe V., Bardou P., Bechner M., Bellec A., Berger A.,
RA Berges H., Bidwell S., Bisseling T., Choisne N., Couloux A., Denny R.,
RA Deshpande S., Dai X., Doyle J.J., Dudez A.M., Farmer A.D., Fouteau S.,
RA Franken C., Gibelin C., Gish J., Goldstein S., Gonzalez A.J., Green P.J.,
RA Hallab A., Hartog M., Hua A., Humphray S.J., Jeong D.H., Jing Y.,
RA Jocker A., Kenton S.M., Kim D.J., Klee K., Lai H., Lang C., Lin S.,
RA Macmil S.L., Magdelenat G., Matthews L., McCorrison J., Monaghan E.L.,
RA Mun J.H., Najar F.Z., Nicholson C., Noirot C., O'Bleness M., Paule C.R.,
RA Poulain J., Prion F., Qin B., Qu C., Retzel E.F., Riddle C., Sallet E.,
RA Samain S., Samson N., Sanders I., Saurat O., Scarpelli C., Schiex T.,
RA Segurens B., Severin A.J., Sherrier D.J., Shi R., Sims S., Singer S.R.,
RA Sinharoy S., Sterck L., Viollet A., Wang B.B., Wang K., Wang M., Wang X.,
RA Warfsmann J., Weissenbach J., White D.D., White J.D., Wiley G.B.,
RA Wincker P., Xing Y., Yang L., Yao Z., Ying F., Zhai J., Zhou L., Zuber A.,
RA Denarie J., Dixon R.A., May G.D., Schwartz D.C., Rogers J., Quetier F.,
RA Town C.D., Roe B.A.;
RT "The Medicago genome provides insight into the evolution of rhizobial
RT symbioses.";
RL Nature 480:520-524(2011).
RN [2] {ECO:0000313|EMBL:AEX20500.1}
RP NUCLEOTIDE SEQUENCE.
RX PubMed=22292047; DOI=10.1371/journal.pone.0030817;
RA Toth K., Stratil T.F., Madsen E.B., Ye J., Popp C., Antolin-Llovera M.,
RA Grossmann C., Jensen O.N., Schussler A., Parniske M., Ott T.;
RT "Functional Domain Analysis of the Remorin Protein LjSYMREM1 in Lotus
RT japonicus.";
RL PLoS ONE 7:E30817-E30817(2012).
RN [3] {ECO:0000313|EMBL:KEH21042.1, ECO:0000313|Proteomes:UP000002051}
RP GENOME REANNOTATION.
RC STRAIN=A17 {ECO:0000313|EMBL:KEH21042.1}, and cv. Jemalong A17
RC {ECO:0000313|EnsemblPlants:KEH21042,
RC ECO:0000313|Proteomes:UP000002051};
RX PubMed=24767513; DOI=10.1186/1471-2164-15-312;
RA Tang H., Krishnakumar V., Bidwell S., Rosen B., Chan A., Zhou S.,
RA Gentzbittel L., Childs K.L., Yandell M., Gundlach H., Mayer K.F.,
RA Schwartz D.C., Town C.D.;
RT "An improved genome release (version Mt4.0) for the model legume Medicago
RT truncatula.";
RL BMC Genomics 15:312-312(2014).
RN [4] {ECO:0000313|EnsemblPlants:KEH21042}
RP IDENTIFICATION.
RC STRAIN=cv. Jemalong A17 {ECO:0000313|EnsemblPlants:KEH21042};
RG EnsemblPlants;
RL Submitted (APR-2015) to UniProtKB.
RN [5] {ECO:0000313|EMBL:RHN43320.1}
RP NUCLEOTIDE SEQUENCE.
RC TISSUE=Leaves {ECO:0000313|EMBL:RHN43320.1};
RA Pecrix Y., Gamas P., Carrere S.;
RT "Whole-genome landscape of Medicago truncatula symbiotic genes.";
RL Nat. Plants 0:0-0(2018).
CC -!- SIMILARITY: Belongs to the remorin family.
CC {ECO:0000256|ARBA:ARBA00005711}.
CC ---------------------------------------------------------------------------
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DR EMBL; JQ061257; AEX20500.1; -; Genomic_DNA.
DR EMBL; CM001224; KEH21042.1; -; Genomic_DNA.
DR EMBL; PSQE01000008; RHN43320.1; -; Genomic_DNA.
DR RefSeq; XP_013447015.1; XM_013591561.1.
DR AlphaFoldDB; H2EST4; -.
DR SwissPalm; H2EST4; -.
DR PaxDb; 3880-AET05022; -.
DR EnsemblPlants; KEH21042; KEH21042; MTR_8g097320.
DR GeneID; 25502038; -.
DR Gramene; KEH21042; KEH21042; MTR_8g097320.
DR KEGG; mtr:25502038; -.
DR OrthoDB; 1107387at2759; -.
DR Proteomes; UP000002051; Chromosome 8.
DR Proteomes; UP000265566; Chromosome 8.
DR ExpressionAtlas; H2EST4; differential.
DR GO; GO:1901700; P:response to oxygen-containing compound; IEA:UniProt.
DR InterPro; IPR005516; Remorin_C.
DR PANTHER; PTHR31775:SF16; CARBOXY-TERMINAL REGION REMORIN; 1.
DR PANTHER; PTHR31775; OS02G0117200 PROTEIN; 1.
DR Pfam; PF03763; Remorin_C; 1.
PE 3: Inferred from homology;
KW Reference proteome {ECO:0000313|Proteomes:UP000002051}.
FT DOMAIN 88..192
FT /note="Remorin C-terminal"
FT /evidence="ECO:0000259|Pfam:PF03763"
FT REGION 1..72
FT /note="Disordered"
FT /evidence="ECO:0000256|SAM:MobiDB-lite"
FT COMPBIAS 38..70
FT /note="Polar residues"
FT /evidence="ECO:0000256|SAM:MobiDB-lite"
SQ SEQUENCE 205 AA; 23339 MW; 55C7C0CE216EA6FD CRC64;
Query Match 100.0%; Score 1027; Length 205;
Best Local Similarity 100.0%;
Matches 205; Conservative 0; Mismatches 0; Indels 0; Gaps 0;
Qy 1 MEESKNKQLELVDTLTPLPQSESEPREFSYFLEEKEPGNEGTSSSVVKQERVVSDHATSS 60
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 1 MEESKNKQLELVDTLTPLPQSESEPREFSYFLEEKEPGNEGTSSSVVKQERVVSDHATSS 60
Qy 61 VDQTTAAGTDTKDSVDRDAVLARVESQKRLALIKAWEENEKTKVENRAYKMQSAVDLWED 120
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 61 VDQTTAAGTDTKDSVDRDAVLARVESQKRLALIKAWEENEKTKVENRAYKMQSAVDLWED 120
Qy 121 DKKASIEAKFKGIEVKLDRKKSEYVEVMQNKIGEIHKSAEEKKAMIEAQKGEEILKVEET 180
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 121 DKKASIEAKFKGIEVKLDRKKSEYVEVMQNKIGEIHKSAEEKKAMIEAQKGEEILKVEET 180
Qy 181 AAKFRTRGYQPRRLLGCFSGLRFFS 205
|||||||||||||||||||||||||
Db 181 AAKFRTRGYQPRRLLGCFSGLRFFS 205
Before the filing date of the instant application, it would have been obvious to one of ordinary skill in the art to modify the teachings of Etzler, Frenzel, and UniProt record A9YWS7 LEC10_MEDTR according to the teachings of Sanz Molinero to also include a nucleotide encoding a remorin protein of instant SEQ ID NO: 25 (claim 27). One would have been motivated to do so because Sanz Molinero teaches that the introduction of a remorin gene led to a significant increase in yield (paragraph [0851], table V). Increasing yield in a transgenic plant would be using a known technique to improve similar products in the same way. Furthermore, one would have had reasonable expectation of success because the remorin gene that successfully increased yield in Sanz Molinero came from Arabidopsis, Sanz Molinero taught a remorin gene in Arabidopsis (At4g00670) that has 100% sequence homology to SEQ ID NO: 25, and transformation methods of legumes were well established at the time of filing.
Before the filing date of the instant application, it would also have been obvious to introduce a homologous SYMREM1 remorin gene from Medicago truncatula as taught by UniProt record H2EST4_MEDTR with 100% identity to SEQ ID NO: 14. Using an orthologous gene from another species would be a design choice to one of ordinary skill in the art. The rationale for adding a polynucleotide segment encoding a SYMREM1 protein of SEQ ID NO: 14 would have been combining prior art elements according to known methods to yield predictable results. The resulting plant would read on both instant claim 9 & 28. Thus, claims 9 & 27-28 are obvious in view of Etzler, Frenzel, UniProt record A9YWS7 LEC10_MEDTR, Sanz Molinero, and UniProt record H2EST4_MEDTR.
Applicant urges that Etzler, Frenzel, UniProt Accession A9YWS7, Molinero, and UniProt Accession H2EST4_MEDTR do not teach or suggest that the LDP1 protein (SEQ ID NO: 2) induces membrane invaginations in the presence of rhizobia and improve symbiotic performance by clustering expressed LDP1 polypeptide, nor do they remedy the mechanistic gap between Etzler’s LNP and LDP1. Applicant urges that the claims provide unexpected and advantageous results which could not have been predicted from the cited references and so are non-obvious (Remarks, page 20, paragraph 3-4 & page 23, paragraph 5-page 24, paragraph 1).
This argument is unpersuasive, because Franzel teaches, based on expression patterns rather than any specific function, that a protein comprising a sequence with 100% identity to SEQ ID NO: 2 is important to mycorrhizal infection. The rationale to arrive at a claimed invention does not need to be Applicant’s own in order to be obvious, so knowledge of the mechanism of membrane invagination was not necessary for one of skill in the art to have known or reasonably expected that a protein with 100% sequence identity to SEQ ID NO: 2 would be important to mycorrhizal symbiotic infection.
A DNA molecule encoding a lectin-domain protein of A9YWS7 LEC10_MEDTR, because it has 100% sequence homology to instant SEQ ID NO: 2, would have been capable of producing membrane invagination when expressed in a plant cell, even if this functionality would not have been a motivation for one of skill in the art to introduce the sequence into a plant cell.
Claim 25 is rejected under 35 U.S.C. 103 as being unpatentable over Etzler et al US 6849777 B1 (filed 9/6/2000, granted 2/1/2005), hereafter Etzler, in view of Frenzel et al (2005) MPMI 18(8):771–782, hereafter Frenzel, and in view of Uniprot record A9YWS7 LEC10_MEDTR (first available 2/5/2008), as applied to claims 1-2, 6-8, & 10-14 above, and further in view of Bono et al. US 2015/0232876 A1 published 8/20/2015, hereafter Bono and Uniprot record A0A396HLA6_MEDTR (first available 12/5/2018).
Due to Applicant' s amendment of the claims, the rejection is modified from the rejection as set forth in the Office action mailed 5/20/2025, as applied to claims 1-3, 6-8, 1-11, 14 & 25. Applicant' s arguments filed 11/18/2025 have been fully considered but they are not persuasive.
Claim 25 is drawn to a plant or plant part comprising a recombinant DNA molecule encoding a protein having the sequence of SEQ ID NO: 2 and further comprising a recombinant DNA molecule encoding a Nod factor perception (NFP) or Lysin Motif Receptor-Like Kinase 3 protein having the sequence of SEQ ID NO: 21.
The teachings of Etzler, Frenzel, and Uniprot record A9YWS7 LEC10_MEDTR are summarized above. They do not teach an NFP protein or SEQ ID NO: 21.
Bono teaches a plant comprising a transgene comprising a polynucleotide encoding a LysM motif receptor kinase (claim 6, lines 1-3), specifically Bono teaches NFP and LYK3 genes inserted into a vector (page 6, paragraph [0077]) under a 35S promoter (page 6, paragraph [0078]) and introduced into N. benthamiana via agroinfiltration (page 6, paragraph [0079]). Bono teaches that the proteins with a LYR3 domain produced by the transgenic N. benthamiana are capable of binding lipochitooligosaccharides (LCOs) (figure 3; page 7, paragraph [0086]), which are signaling molecules necessary for symbiotic association between plants and Rhizobia or arbuscular mycorrhizal fungi (page 1, paragraph [0006]). Bono teaches a motivation for using proteins from M. truncatula because M. truncatula establishes rhizobial symbiosis or symbiotic arbuscular endomycorrhiza and other symbiosis-related proteins are rich in binding sites (page 5, paragraph [0059]). Bono teaches that NFP and LYK3 proteins are involved in the perception of Nod factors and Myc factors (page 1, paragraph [0016] and furthermore teaches that Nod factors have a beneficial effect on the growth and development of plants in addition to playing a role in symbiosis (page 1, paragraph [0009]), thus providing a motivation to introduce NFP to perceive Nod factors into plants.
Uniprot record A0A396HLA6_MEDTR teaches an NFP protein having 100% sequence identity to SEQ ID NO: 21. See alignment below.
A0A396HLA6_MEDTR
ID A0A396HLA6_MEDTR Unreviewed; 595 AA.
AC A0A396HLA6;
DT 05-DEC-2018, integrated into UniProtKB/TrEMBL.
DT 05-DEC-2018, sequence version 1.
DT 27-MAR-2024, entry version 18.
DE SubName: Full=Putative serine/threonine receptor-like kinase NFP RLK-Pelle-LysM family {ECO:0000313|EMBL:RHN54132.1};
DE EC=2.7.10.- {ECO:0000313|EMBL:RHN54132.1};
GN ORFNames=MtrunA17_Chr5g0403371 {ECO:0000313|EMBL:RHN54132.1};
OS Medicago truncatula (Barrel medic) (Medicago tribuloides).
OC Eukaryota; Viridiplantae; Streptophyta; Embryophyta; Tracheophyta;
OC Spermatophyta; Magnoliopsida; eudicotyledons; Gunneridae; Pentapetalae;
OC rosids; fabids; Fabales; Fabaceae; Papilionoideae; 50 kb inversion clade;
OC NPAAA clade; Hologalegina; IRL clade; Trifolieae; Medicago.
OX NCBI_TaxID=3880 {ECO:0000313|EMBL:RHN54132.1};
RN [1] {ECO:0000313|EMBL:RHN54132.1}
RP NUCLEOTIDE SEQUENCE.
RC TISSUE=Leaves {ECO:0000313|EMBL:RHN54132.1};
RA Pecrix Y., Gamas P., Carrere S.;
RT "Whole-genome landscape of Medicago truncatula symbiotic genes.";
RL Nat. Plants 0:0-0(2018).
CC -!- CAUTION: The sequence shown here is derived from an EMBL/GenBank/DDBJ
CC whole genome shotgun (WGS) entry which is preliminary data.
CC {ECO:0000313|EMBL:RHN54132.1}.
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DR EMBL; PSQE01000005; RHN54132.1; -; Genomic_DNA.
DR RefSeq; XP_003611889.1; XM_003611841.2.
DR AlphaFoldDB; A0A396HLA6; -.
DR SMR; A0A396HLA6; -.
DR GeneID; 11410973; -.
DR KEGG; mtr:11410973; -.
DR OMA; SPPSCET; -.
DR OrthoDB; 923763at2759; -.
DR Proteomes; UP000265566; Chromosome 5.
DR GO; GO:0016020; C:membrane; IEA:UniProtKB-KW.
DR GO; GO:0005524; F:ATP binding; IEA:InterPro.
DR GO; GO:0004672; F:protein kinase activity; IEA:InterPro.
DR GO; GO:0016310; P:phosphorylation; IEA:UniProtKB-KW.
DR Gene3D; 1.10.510.10; Transferase(Phosphotransferase) domain 1; 2.
DR InterPro; IPR011009; Kinase-like_dom_sf.
DR InterPro; IPR000719; Prot_kinase_dom.
DR InterPro; IPR001245; Ser-Thr/Tyr_kinase_cat_dom.
DR InterPro; IPR008266; Tyr_kinase_AS.
DR PANTHER; PTHR45927; LYSM-DOMAIN RECEPTOR-LIKE KINASE-RELATED; 1.
DR PANTHER; PTHR45927:SF2; SERINE_THREONINE RECEPTOR-LIKE KINASE NFP; 1.
DR Pfam; PF07714; PK_Tyr_Ser-Thr; 1.
DR SUPFAM; SSF56112; Protein kinase-like (PK-like); 1.
DR PROSITE; PS50011; PROTEIN_KINASE_DOM; 1.
DR PROSITE; PS00109; PROTEIN_KINASE_TYR; 1.
PE 4: Predicted;
KW Kinase {ECO:0000313|EMBL:RHN54132.1}; Membrane {ECO:0000256|SAM:Phobius};
KW Receptor {ECO:0000313|EMBL:RHN54132.1}; Signal {ECO:0000256|SAM:SignalP};
KW Transferase {ECO:0000313|EMBL:RHN54132.1};
KW Transmembrane {ECO:0000256|SAM:Phobius};
KW Transmembrane helix {ECO:0000256|SAM:Phobius}.
FT SIGNAL 1..27
FT /evidence="ECO:0000256|SAM:SignalP"
FT CHAIN 28..595
FT /evidence="ECO:0000256|SAM:SignalP"
FT /id="PRO_5017213836"
FT TRANSMEM 245..271
FT /note="Helical"
FT /evidence="ECO:0000256|SAM:Phobius"
FT DOMAIN 284..573
FT /note="Protein kinase"
FT /evidence="ECO:0000259|PROSITE:PS50011"
SQ SEQUENCE 595 AA; 66014 MW; 5E52FF7A8AA1F0EA CRC64;
Query Match 100.0%; Score 3029; Length 595;
Best Local Similarity 100.0%;
Matches 595; Conservative 0; Mismatches 0; Indels 0; Gaps 0;
Qy 1 MSAFFLPSSSHALFLVLMLFFLTNISAQPLYISETNFTCPVDSPPSCETYVAYRAQSPNF 60
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 1 MSAFFLPSSSHALFLVLMLFFLTNISAQPLYISETNFTCPVDSPPSCETYVAYRAQSPNF 60
Qy 61 LSLSNISDIFNLSPLRIAKASNIEAEDKKLIPDQLLLVPVTCGCTKNHSFANITYSIKQG 120
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 61 LSLSNISDIFNLSPLRIAKASNIEAEDKKLIPDQLLLVPVTCGCTKNHSFANITYSIKQG 120
Qy 121 DNFFILSITSYQNLTNYLEFKNFNPNLSPTLLPLDTKVSVPLFCKCPSKNQLNKGIKYLI 180
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 121 DNFFILSITSYQNLTNYLEFKNFNPNLSPTLLPLDTKVSVPLFCKCPSKNQLNKGIKYLI 180
Qy 181 TYVWQDNDNVTLVSSKFGASQVEMLAENNHNFTASTNRSVLIPVTSLPKLDQPSSNGRKS 240
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 181 TYVWQDNDNVTLVSSKFGASQVEMLAENNHNFTASTNRSVLIPVTSLPKLDQPSSNGRKS 240
Qy 241 SSQNLALIIGISLGSAFFILVLTLSLVYVYCLKMKRLNRSTSSSETADKLLSGVSGYVSK 300
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 241 SSQNLALIIGISLGSAFFILVLTLSLVYVYCLKMKRLNRSTSSSETADKLLSGVSGYVSK 300
Qy 301 PTMYEIDAIMEGTTNLSDNCKIGESVYKANIDGRVLAVKKIKKDASEELKILQKVNHGNL 360
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 301 PTMYEIDAIMEGTTNLSDNCKIGESVYKANIDGRVLAVKKIKKDASEELKILQKVNHGNL 360
Qy 361 VKLMGVSSDNDGNCFLVYEYAENGSLEEWLFSESSKTSNSVVSLTWSQRITIAMDVAIGL 420
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 361 VKLMGVSSDNDGNCFLVYEYAENGSLEEWLFSESSKTSNSVVSLTWSQRITIAMDVAIGL 420
Qy 421 QYMHEHTYPRIIHRDITTSNILLGSNFKAKIANFGMARTSTNSMMPKIDVFAFGVVLIEL 480
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 421 QYMHEHTYPRIIHRDITTSNILLGSNFKAKIANFGMARTSTNSMMPKIDVFAFGVVLIEL 480
Qy 481 LTGKKAMTTKENGEVVILWKDFWKIFDLEGNREERLRKWMDPKLESFYPIDNALSLASLA 540
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 481 LTGKKAMTTKENGEVVILWKDFWKIFDLEGNREERLRKWMDPKLESFYPIDNALSLASLA 540
Qy 541 VNCTADKSLSRPTIAEIVLCLSLLNQPSSEPMLERSLTSGLDAEATHVVTSVIAR 595
|||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 541 VNCTADKSLSRPTIAEIVLCLSLLNQPSSEPMLERSLTSGLDAEATHVVTSVIAR 595
Before the time of filing of the instant application, it would have been obvious to one having ordinary skill in the art to modify the teachings of Etzler, Frenzel, and Uniprot record A9YWS7 LEC10_MEDTR with the teachings of Bono to create a plant comprising a recombinant DNA molecule encoding an NFP protein. One of ordinary skill would have been motivated to combine the teachings because Bono teaches that transforming a plant with a nucleotide expressing a protein with a LysM motif improves plant response to LCOs (Bono claim 2, lines 1-7) and that recognition of LCOs has a beneficial effect on growth of plants and on symbiosis. One having ordinary skill in the art would have been reasonably confident of success because plant transformation was routine in the art, and both proteins were suggested to be important for symbiosis.
Before the time of filing of the instant application, it would also have been obvious to use a nucleotide encoding the known protein sequence of Uniprot record A0A396HLA6_MEDTR. Substituting one known sequence of a protein for another known sequence of a homologous protein would have been obvious to one of ordinary skill to produce reasonable expectation of success. Because the protein of Uniprot record A0A396HLA6_MEDTR was known to be an NFP, a polynucleotide segment encoding this protein would be a polynucleotide segment encoding an NFP.
Applicant urges that Bono and UniProt Record A0A396HLA6_MEDTR do not teach or suggest that SEQ ID NO: 2 induces membrane invaginations in the presence of rhizobia and improve symbiotic performance by clustering expressed LDP1 polypeptide at the cellular bilayer so they do not cure the deficiency of Etzler, Frenzel, or UniProt A9YWS7. Thus, Applicant urges the instant claims provide unexpected and advantageous results which could not have been predicted from the cited references (Remarks, page 21, paragraphs 4-5).
This argument is unpersuasive, because Franzel teaches, based on expression patterns rather than any specific function, that a protein comprising a sequence with 100% identity to SEQ ID NO: 2 is important to mycorrhizal infection. The rationale to arrive at a claimed invention does not need to be Applicant’s own in order to be obvious, so knowledge of the mechanism of membrane invagination was not necessary for one of skill in the art to have known or reasonably expected that a protein with 100% sequence identity to SEQ ID NO: 2 would be important to mycorrhizal symbiotic infection.
A DNA molecule encoding a lectin-domain protein of A9YWS7 LEC10_MEDTR, because it has 100% sequence homology to instant SEQ ID NO: 2, would have been capable of producing membrane invagination when expressed in a plant cell, even if this functionality would not have been a motivation for one of skill in the art to introduce the sequence into a plant cell.
Claim 26 is rejected under 35 U.S.C. 103 as being unpatentable over Etzler et al US 6849777 B1 (filed 9/6/2000, granted 2/1/2005), hereafter Etzler, Frenzel et al (2005) MPMI 18(8):771–782, hereafter Frenzel, and Uniprot record A9YWS7 LEC10_MEDTR (first available 2/5/2008) in view of Bono et al. US 2015/0232876 A1 published 8/20/2015, hereafter Bono and UniProt record A0A396HLA6_MEDTR (first available 12/5/2018), as applied to claims 1-2, 6-8, 10-14 & 25 above, and further in view of Sanz Molinero et al (US 20100199379 A1, published 8/5/2010), hereafter Sanz Molinero; and UniProt record H2EST4_MEDTR (first available 3/21/2012).
Due to Applicant' s amendment of the claims, the rejection is modified from the rejection as set forth in the Office action mailed 5/20/2025, as applied to claims 1-3, 6-8, 10-14 & 25-26. Applicant' s arguments filed 11/18/2025 have been fully considered but they are not persuasive.
Claim 26 is drawn to the plant of claim 25 further comprising a DNA molecule comprising a heterologous promoter and encoding a SYMREM1 protein with the sequence of SEQ ID NO: 14.
The teachings of Etzler, Frenzel, Uniprot record A9YWS7 LEC10_MEDTR, Bono, and Uniprot record A0A396HLA6_MEDTR (first available 12/5/2018) are summarized above. They do not teach a recombinant DNA molecule comprising a promoter operably linked to a nucleotide encoding a protein of SEQ ID NO: 14.
The teachings of Sanz Molinero and UniProt record H2EST4_MEDTR are presented above.
Before the filing date of the instant application, it would have been obvious to one of ordinary skill in the art to modify the teachings of Etzler, Frenzel, UniProt record A9YWS7 LEC10_MEDTR, Bono, and UniProt record A0A396HLA6_MEDTR according to the teachings of Sanz Molinero to also include a nucleotide encoding a remorin protein. One would have been motivated to do so because Sanz Molinero teaches that the introduction of a remorin gene led to a significant increase in yield in transgenic plants. Increased yield in a crop plant is an obvious improvement, and increasing yield in a transgenic plant would be use of known technique to improve similar products in the same way. Furthermore, one of ordinary skill would have had reasonable expectation of success because transformation methods of legumes were well established at the time of filing.
Before the filing date of the instant application, it would have been obvious to substitute an Arabidopsis remorin gene as in Sanz Molinero for a homologous SYMREM1 remorin gene from Medicago truncatula as taught by UniProt record H2EST4_MEDTR. Using an orthologous gene from another species would be a design choice to one of ordinary skill in the art. Thus, claim 26 is obvious in view of Etzler, Frenzel, UniProt record A9YWS7 LEC10_MEDTR, Bono, UniProt record A0A396HLA6_MEDTR, Sanz Molinero, and UniProt record H2EST4_MEDTR.
Applicant urges that Etzler, Frenzel, UniProt Accession A9YWS7, UniProt Record A0A396HLA6_MEDTR, Molinero, and UniProt A0A396HLA6_MEDTR do not teach that a protein of SEQ ID NO: 2 can induce membrane invaginations in the presence of rhizobia and improve symbiotic performance by clustering expressed LDP1 at the cellular bilayer and do not bridge the mechanistic divide between an ecto-apyrase and LDP1’s curvature-drive infection-structure-localized function. Because of this, Applicant urges the instant claims provide unexpected and advantageous results that make the claims non-obvious (Remarks, page 22, paragraph 5-page 23, paragraph 1).
This argument is unpersuasive, because Franzel teaches, based on expression patterns rather than any specific function, that a protein comprising a sequence with 100% identity to SEQ ID NO: 2 is important to mycorrhizal infection. The rationale to arrive at a claimed invention does not need to be Applicant’s own in order to be obvious, so knowledge of the mechanism of membrane invagination was not necessary for one of skill in the art to have known or reasonably expected that a protein with 100% sequence identity to SEQ ID NO: 2 would be important to mycorrhizal symbiotic infection.
A DNA molecule encoding a lectin-domain protein of A9YWS7 LEC10_MEDTR, because it has 100% sequence homology to instant SEQ ID NO: 2, would have been capable of producing membrane invagination when expressed in a plant cell, even if this functionality would not have been a motivation for one of skill in the art to introduce the sequence into a plant cell.
Claim 15 is rejected under 35 U.S.C. 103 as being unpatentable over Etzler, Frenzel, and Uniprot record A9YWS7 LEC10_MEDTR, as applied to claims 1,2, 6-8 & 10-14 above and further in view of Vershinina et al (2012). Symbiosis. 56:25–33, (published online 2/15/2012, hereafter Vershinina).
Due to Applicant' s amendment of the claims, the rejection is modified from the rejection as set forth in the Office action mailed 5/20/2025, as applied to claims 1-3, 6-8, 10-15. Applicant' s arguments filed 11/18/2025 have been fully considered but they are not persuasive.
Claim 15 is drawn to a method for increasing symbiotic infection in a plant compared to an untreated plant comprising obtaining a plant comprising a cell comprising a lectin-domain containing protein of the sequence of SEQ ID NO: 2, and contacting said plant with an effective amount of rhizobia bacterium.
The teachings of Etzler, Frenzel, and Uniprot record A9YWS7 LEC10_MEDTR are presented above. Etzler teaches a method for increasing symbiotic infection in a plant comprising introducing a lectin-domain containing protein in a plant, in an expression cassette. Furthermore, Etzler teaches a method comprising the step of infecting the plants with mycorrhizal fungi. Etzler, Frenzel, and Uniprot record A9YWS7 LEC10_MEDTR do not teach a step of contacting a plant with an effective amount of one or more rhizobia bacterium.
Vershinina teaches a method of transforming tobacco, tomato, and rape with a gene encoding a lectin-domain containing protein, psl (page 31, left column, lines 7-28). Vershinina teaches that lectins are important for the attachment of rhizobia to host-plant roots (page 26, left column, lines 1-7 and lines 14-22). Vershinina teaches that lectin is a tool to help in using rhizobia to improve the growth of non-legumes (page 26, left column, lines 21-35). Vershinina teaches inoculating the transgenic plants with Rhizobium leguminosarum and Rhizobium galegae, leading to R. leguminosarum bacteria adhering to the plant roots (page 29, figure 4).
Before the time of filing of the instant application, it would have been obvious to one of ordinary skill in the art to substitute Etzler’s step of infecting with a mycorrhizal fungus with the inoculation of plant roots with Rhizobium bacteria to cause adherence of bacteria on roots, as taught by Vershinina. One of ordinary skill in the art would have been motivated to substitute Rhizobium bacteria for a mycorrhizal fungus in order to increase nodulation, because Etzler had taught that lectin proteins are involved in Rhizobia association as well as mycorrhizal association and nodulation. One of ordinary skill in the art would have had reasonable expectation of success, because transformation of plants was routine in the art and Vershinina had shown that expression of a lectin transgene led to rhizobial bacteria adhering even to non-legume plant roots.
Thus, the method of claim 15 is obvious in view of Etzler, Frenzel, UniProt record A9YWS7 LEC10_MEDTR, and Vershinina.
Applicant urges that Etzler, Frenzel, UniProt Accession A9YWS7, Bono, and UniProt Accession A0A396HLA6_MEDTR do not teach that SEQ ID NO: 2 can induce membrane invaginations and improve symbiotic performance by clustering expressed LDP1 at the cellular bilayer. Applicant urges that Vershinina focuses on adherence and so does not suggest the curvature-drive localized mechanism of LDP1, so the results of the instant claims are non-obvious (Remarks, page 24, paragraph 5-page 25, paragraph 1).
This argument is unpersuasive, because Franzel teaches, based on expression patterns rather than any specific function, that a protein comprising a sequence with 100% identity to SEQ ID NO: 2 is important to mycorrhizal infection. The rationale to arrive at a claimed invention does not need to be Applicant’s own in order to be obvious, so knowledge of the mechanism of membrane invagination was not necessary for one of skill in the art to have known or reasonably expected that a protein with 100% sequence identity to SEQ ID NO: 2 would be important to mycorrhizal symbiotic infection.
A DNA molecule encoding a lectin-domain protein of A9YWS7 LEC10_MEDTR, because it has 100% sequence homology to instant SEQ ID NO: 2, would have been capable of producing membrane invagination when expressed in a plant cell, even if this functionality would not have been a motivation for one of skill in the art to introduce the sequence into a plant cell.
Claim 16 is rejected under 35 U.S.C. 103 as being unpatentable over Etzler, Frenzel, Uniprot record A9YWS7 LEC10_MEDTR, and Vershinina as applied to claims 1,2, 6-8, 10-15 above, and further in view of Schmeisser et al (2009). Applied and Environmental Microbiology. 75(12), hereafter Schmeisser.
Due to Applicant' s amendment of the claims, the rejection is modified from the rejection as set forth in the Office action mailed 5/20/2025, as applied to claims 1-3, 6-8, 10-16. Applicant' s arguments filed 11/18/2025 have been fully considered but they are not persuasive.
Claim 16 is drawn to the method of claim 15 wherein the rhizobia bacterium is Rhizobium sp IRBG74 or NG234.
The teachings of Etzler, Frenzel, UniProt record A9WS7 LEC10_MEDTR, and Vershinina are presented above. These references do not teach associating a plant with Rhizobium sp. IRBG74 or NGR234 specifically.
Schmeisser (published 6/15/2009) teaches that Rhizobium sp. Strain NGR234 is a versatile bacterium capable of forming nodules with more legumes than other microsymbionts (abstract). Thus, Schmeisser teaches a motivation for using Rhizobium sp. Strain NGR234 in a method for increasing symbiotic infection in a plant.
Before the filing of the instant application, it would have been obvious to one of ordinary skill in the art to substitute the R. leguminosarum of Vershinina with Rhizobium sp. NGR234. One of ordinary skill in the art would have been motivated to do so because Rhizobium sp. NGR234 was known to form symbiotic nodules with more legumes than any other microsymbiont, making this a versatile bacterium to use. One of ordinary skill would have had reasonable expectation of success, because Rhizobium sp. NGR234 forms symbiotic nodules many legumes.
Applicant urges that Etzler, Frenzel, UniProt Accession A9YWS7, Vershinina do not teach or suggest that SEQ ID NO: 2 induces membrane invaginations and improve symbiotic performance by clustering expressed LDP1 at the cellular bilayer and Schmeisser does not cure this deficiency, so the results of the instant claims are unexpectedly advantageous and non-obvious (Remarks, page 25, paragraph 5-page 26, paragraph 1).
This argument is unpersuasive, because Franzel teaches, based on expression patterns rather than any specific function, that a protein comprising a sequence with 100% identity to SEQ ID NO: 2 is important to mycorrhizal infection. The rationale to arrive at a claimed invention does not need to be Applicant’s own in order to be obvious, so knowledge of the mechanism of membrane invagination was not necessary for one of skill in the art to have known or reasonably expected that a protein with 100% sequence identity to SEQ ID NO: 2 would be important to mycorrhizal symbiotic infection.
A DNA molecule encoding a lectin-domain protein of A9YWS7 LEC10_MEDTR, because it has 100% sequence homology to instant SEQ ID NO: 2, would have been capable of producing membrane invagination when expressed in a plant cell, even if this functionality would not have been a motivation for one of skill in the art to introduce the sequence into a plant cell.
Claim 17 is rejected under 35 U.S.C. 103 as being unpatentable over Etzler, Frenzel, Uniprot record A9YWS7 LEC10_MEDTR, and Vershinina, as applied to claims 1,2, 6-8, 10-15 above, and further in view of Sanz Molinero et al (US 20100199379 A1, published 8/5/2010), hereafter Sanz Molinero and UniProt record H2EST4_MEDTR (first available 3/21/2012).
This is a new rejection from the rejection as set forth in the Office action mailed 5/20/2025. Applicant' s arguments filed 11/18/2025 have been fully considered as they apply to the new rejection, but they are not persuasive.
Claim 17 is drawn to the method of claim 15 comprising expressing a protein with the sequence of SEQ ID NO: 14 in a plant.
The teachings of Vershinina, Frenzel, and UniProt record A9YWS7 LEC10_MEDTR are presented above. These references do not teach a protein of SEQ ID NO: 14 expressed in a plant.
The teachings of Sanz Molinero and UniProt record H2EST4_MEDTR teaching a remorin protein expressed in a plant and a remorin gene with 100% sequence identity to SEQ ID NO: 14 are also presented above.
Before the filing date of the instant application, it would have been obvious to one of ordinary skill in the art to modify the teachings of Etzler, Frenzel, UniProt record A9YWS7 LEC10_MEDTR, and Vershinina according to the teachings of Sanz Molinero to also include a nucleotide encoding a remorin protein. One would have been motivated to do so because Sanz Molinero teaches that the introduction of a remorin gene led to a significant increase in yield. Furthermore, one would have had reasonable expectation of success because transformation methods of legumes were well established at the time of filing.
Before the filing date of the instant application, it would have been obvious to substitute an Arabidopsis remorin gene as in Sanz Molinero for a homologous remorin gene from Medicago truncatula as taught by UniProt record H2EST4_MEDTR. Using an orthologous gene from another species would be a design choice to one of ordinary skill in the art. Thus, claim 17 is obvious in view of Etzler, Frenzel, UniProt record A9YWS7 LEC10_MEDTR, Vershinina, Sanz Molinero, and UniProt record H2EST4_MEDTR.
Applicant urges that Etzler, Frenzel, UniProt Accession A9YWS7, Vershinina, and Schmeisser do not teach or suggest that SEQ ID NO: 2 induces membrane invaginations and improve symbiotic performance by clustering expressed LDP1 at the cellular bilayer and Sanz Molinero and UniProt Accession H2EST4_MEDTR do not cure this deficiency. Applicant urges that substituting LDP1 with apyrase LNP is unsupported mechanistically and structurally. Applicant urges that the results of the instant claims are unexpectedly advantageous and non-obvious (Remarks, page 26, paragraph 5-page 27, paragraph 1).
This argument is unpersuasive, because Franzel teaches, based on expression patterns rather than any specific function, that a protein comprising a sequence with 100% identity to SEQ ID NO: 2 is important to mycorrhizal infection. The rationale to arrive at a claimed invention does not need to be Applicant’s own in order to be obvious, so knowledge of the mechanism of membrane invagination was not necessary for one of skill in the art to have known or reasonably expected that a protein with 100% sequence identity to SEQ ID NO: 2 would be important to mycorrhizal symbiotic infection.
The obviousness of providing a plant cell with a transgene encoding a lectin protein in order to increase symbiosis is further demonstrated by Vershinina, because the psl lectin protein taught by Vershinina does not comprise apyrase activity and yet still increases adherence of rhizobial bacteria to plant roots. One of skill in the art would have understood prior to the filing of the instant application that proteins containing lectin-domains may be involved in symbiotic infection even without apyrase activity.
A DNA molecule encoding a lectin-domain protein of A9YWS7 LEC10_MEDTR, because it has 100% sequence homology to instant SEQ ID NO: 2, would have been capable of producing membrane invagination when expressed in a plant cell, even if this functionality would not have been a motivation for one of skill in the art to introduce the sequence into a plant cell. Thus, the use of a lectin-domain containing protein taught to be important to mycorrhizal infection according to the instant claims would not have been non-obvious.
Conclusion
No claims are allowed.
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/VICTORIA L DELEO/Examiner, Art Unit 1662
/Anne Kubelik/Primary Examiner, Art Unit 1663