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 .
Claim Status
Claims 1 and 13-27 are pending.
Claims 22-27 are not examined as part of non-elected group.
Claims 1 and 13-21 are elected and are being examined.
All previous objections and rejections not set forth below have been withdrawn in view .of applicant’s amendments to the claims.
Priority
Acknowledgment is made of applicant’s claim for foreign priority under 35 U.S.C. 119 (a)-(d). The certified copy has been filed in parent Application No. CN 202211137305.8, filed on 12/08/2023.
Claim Rejections - 35 USC § 103
Claims 1 and 13 are rejected under 35 U.S.C. 103 as being unpatentable over Ma et al. in view of Cao et al. and DeBlasio et al.
Claim 1 is drawn to a method for obtaining a plant capable of resisting a pathogen by reducing or inhibiting Flotillin1 and Importin α4 genes in the plant.
Ma et al. describes a mechanism underlying movement of pathogenic viral RNAs (as recited in claim 13) including the single stranded RNA of potato spindle tuber viroid (PSTVd) and other RNA molecules (i.e. small RNAs, tRNAs, and rRNAs) from cytoplasm into the nucleus in plants (page 3543, left column, line 5-7). It describes Importin α4 (IMPα-4) being involved in viroid RNA nuclear import, and thus, the involvement of Importin-based cellular pathway in RNA nuclear import (abstract). Reducing the amount of IMP α4 in cells inhibits PSTVd nuclear accumulation and infectivity (page 3544, para 3 “findings”, line 5-6).
However, Ma et al. does not describe Flotillin1 protein or gene encoding the protein.
Cao et al. describes that Flotillin1 (Flot1) is involved in endocytosis (page 2, para 1, line 12-13; page 2, last para, last 2 lines) which is long known to play a crucial part in diverse biological processes including response to pathogen attack. Pathogens enter specific host organisms including plants via endocytosis. Reduced expression/activity of Flot1 in Flot1 amiRNAi mutants in Arabidopsis displays defects in immune responses against (bacterial) pathogens (page 9, para 3, line 20-21). This indicates that reduced expression of Flot1 prevents the pathogen-bacteria from entering the cell and establishing infection.
DeBlasio et al. describes isolation of viral pathogen (Potato Leafroll Virus, PLRV) RNA (as recited in claim 13) interacting with the plant host proteins from systemically infected potato plants (abstract, line 2-4). It describes identifying a finite number of 44 potato proteins interacting with the viral RNA (abstract, line 5). One of the identified host proteins interacting with the viral RNA is flotillin1 (page 4605, right column, para 2, line 21; page 4607, Table 1; and Supplemental Table S1). This indicates that reduced expression of flotillin1 prevents the pathogen-virus from entering the cell and establishing infection.
Before the effective filing date of the invention, it would have been obvious to an ordinarily skilled artisan to inhibit or reduce expression level of the Importin α4 (as described by Ma et al.) and flotillin1 genes (as described by Cao et al. and DeBlasio et al.) with a realistic goal to obtain a plant capable to resist viral pathogen(s).
Before the effective filing date, an ordinarily skilled artisan would have been motivated to inhibit or reduce expression level of the Importin α4 and flotillin1 genes in a plant with a realistic goal to obtain a plant capable to resist both bacterial and viral pathogen(s). Stackability of resistance to diverse groups of pathogens is known and desirable trait in the art.
Claims 14-21 are rejected under 35 U.S.C. 103 as being unpatentable over Ma et al. in view of Cao et al. and DeBlasio et al. as applied to claims 1 and 13 above, and further in view of Kawahara et al.
Claim 14 depends from claim 13 and is drawn to a method comprising one or more characteristics selected from the group consisting of the following items: (i) the plant is selected from the group consisting of wheat, barley, corn, rice, sorghum; (ii) the pathogen is a rice stripe virus; (iii) the protein encoded by the Importin α4 gene has an amino acid sequence as shown in SEQ ID NO: 3; and, (iv) the protein encoded by the Flotillin1 gene has an amino acid sequence as shown in SEQ ID NO: 1.
Ma et al. in view of Cao et al. and DeBlasio et al. describes reducing expression level of the Importin α4 and flotillin1 genes in a plant to obtain a plant capable to resist viral pathogen(s), as discussed above. Ma et al. also describes transforming a binary vector into agrobacterium (page 3553, left column, para 1, line 8-10) followed by agrobacterium mediated transformation and involvement of Importin α4 in nuclear import of T-DNA (page 3546, Fig. 1; page 3552, left column, para 2, line 13-15), as recited in claim 18.
However, Ma et al. in view of Cao et al. and DeBlasio et al. does not describe any Flotillin1 or Importin α4 gene in rice, wheat, barley, corn, or sorghum.
Kawahara et al. teaches rice genes encoding Flotillin1 (UniPort accession No. Q9AV57) and Importin α4 (UniPort accession No. Q0D3F8) proteins. The rice Flotillin1 gene encodes a protein having 100% sequence identity to instant SEQ ID NO: 1, as shown below.
RESULT 1
FLOT1_ORYSJ
ID FLOT1_ORYSJ Reviewed; 485 AA.
AC Q9AV57; A0A0P0XW05;
DT 13-JUL-2010, integrated into UniProtKB/Swiss-Prot.
DT 01-JUN-2001, sequence version 1.
DT 18-JUN-2025, entry version 113.
DE RecName: Full=Flotillin-like protein 1;
DE AltName: Full=Nodulin-like protein 1;
GN Name=FLOT1; OrderedLocusNames=Os10g0481500, LOC_Os10g34040;
GN ORFNames=OsJ_31920, OSJNBa0012L23.36, OSJNBa0093B11.7;
OS Oryza sativa subsp. japonica (Rice).
OC Eukaryota; Viridiplantae; Streptophyta; Embryophyta; Tracheophyta;
OC Spermatophyta; Magnoliopsida; Liliopsida; Poales; Poaceae; BOP clade;
OC Oryzoideae; Oryzeae; Oryzinae; Oryza; Oryza sativa.
OX NCBI_TaxID=39947;
RP GENOME REANNOTATION.
RC STRAIN=cv. Nipponbare;
RX PubMed=24280374; DOI=10.1186/1939-8433-6-4;
RA Kawahara Y., de la Bastide M., Hamilton J.P., Kanamori H., McCombie W.R.,
RA Ouyang S., Schwartz D.C., Tanaka T., Wu J., Zhou S., Childs K.L.,
RA Davidson R.M., Lin H., Quesada-Ocampo L., Vaillancourt B., Sakai H.,
RA Lee S.S., Kim J., Numa H., Itoh T., Buell C.R., Matsumoto T.;
RT "Improvement of the Oryza sativa Nipponbare reference genome using next
RT generation sequence and optical map data.";
RL Rice 6:4-4(2013).
DR EMBL; AC024594; AAK21341.1; -; Genomic_DNA.
DR EMBL; AC051632; AAM91865.1; -; Genomic_DNA.
DR EMBL; DP000086; AAP54307.1; -; Genomic_DNA.
DR EMBL; AP008216; BAF26794.1; -; Genomic_DNA.
DR EMBL; AP014966; BAT11345.1; -; Genomic_DNA.
DR EMBL; CM000147; EAZ16450.1; -; Genomic_DNA.
DR RefSeq; XP_015613363.1; XM_015757877.1.
DR AlphaFoldDB; Q9AV57; -.
DR SMR; Q9AV57; -.
DR FunCoup; Q9AV57; 38.
DR STRING; 39947.Q9AV57; -.
DR PaxDb; 39947-Q9AV57; -.
DR EnsemblPlants; Os10t0481500-00; Os10t0481500-00; Os10g0481500.
DR Gramene; Os10t0481500-00; Os10t0481500-00; Os10g0481500.
DR KEGG; dosa:Os10g0481500; -.
DR eggNOG; KOG2668; Eukaryota.
DR HOGENOM; CLU_030844_1_1_1; -.
DR InParanoid; Q9AV57; -.
DR OMA; IHDQTGM; -.
DR OrthoDB; 6080404at2759; -.
DR Proteomes; UP000000763; Chromosome 10.
DR Proteomes; UP000007752; Chromosome 10.
DR Proteomes; UP000059680; Chromosome 10.
DR GO; GO:0005901; C:caveola; IEA:UniProtKB-SubCell.
DR GO; GO:0005886; C:plasma membrane; IBA:GO_Central.
DR GO; GO:0044853; C:plasma membrane raft; IBA:GO_Central.
DR CDD; cd03399; SPFH_flotillin; 1.
DR Gene3D; 3.30.479.30; Band 7 domain; 1.
DR InterPro; IPR001107; Band_7.
DR InterPro; IPR036013; Band_7/SPFH_dom_sf.
DR InterPro; IPR027705; Flotillin_fam.
DR PANTHER; PTHR13806:SF31; FLOTILLIN-LIKE PROTEIN 1-RELATED; 1.
DR PANTHER; PTHR13806; FLOTILLIN-RELATED; 1.
DR Pfam; PF01145; Band_7; 1.
DR SUPFAM; SSF117892; Band 7/SPFH domain; 1.
PE 3: Inferred from homology;
KW Cell membrane; Coiled coil; Lipoprotein; Membrane; Palmitate;
KW Reference proteome.
FT CHAIN 1..485
FT /note="Flotillin-like protein 1"
FT /id="PRO_0000395212"
FT COILED 228..320
FT /evidence="ECO:0000255"
FT LIPID 38
FT /note="S-palmitoyl cysteine"
FT /evidence="ECO:0000255"
SQ SEQUENCE 485 AA; 52987 MW; A52FB4D0343FA5DC CRC64;
Query Match 100.0%; Score 2434; Length 485; Best Local Similarity 100.0%;
Matches 485; Conservative 0; Mismatches 0; Indels 0; Gaps 0;
Qy 1 MGFAYRIASASEYLAITGYGIADVKLAKKAWVAPGQRCTRFDISPVNYTFEVQAMSAEKL 60
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 1 MGFAYRIASASEYLAITGYGIADVKLAKKAWVAPGQRCTRFDISPVNYTFEVQAMSAEKL 60
Qy 61 PFILPAVFTIGPRADDDDCLLRYAKLISPHDKLSHHVNELVKGVIEGETRVLAASMTMEE 120
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 61 PFILPAVFTIGPRADDDDCLLRYAKLISPHDKLSHHVNELVKGVIEGETRVLAASMTMEE 120
Qy 121 IFQGTKSFKQAVFENVQLELNQFGLIIYNANVKQLVDVAGHEYFSYLGQKTQQEAVNQAK 180
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 121 IFQGTKSFKQAVFENVQLELNQFGLIIYNANVKQLVDVAGHEYFSYLGQKTQQEAVNQAK 180
Qy 181 VDVAEARMKGEVGAKERDGMTRQNAAKVDAETKVYTVKRQGEGAKEEARVKAEVKVFENE 240
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 181 VDVAEARMKGEVGAKERDGMTRQNAAKVDAETKVYTVKRQGEGAKEEARVKAEVKVFENE 240
Qy 241 REAEVAEANADLAMKKAGWQRQAMVAEVEAAKAVAIREAELQVEVERTNASRQTEKLKAE 300
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 241 REAEVAEANADLAMKKAGWQRQAMVAEVEAAKAVAIREAELQVEVERTNASRQTEKLKAE 300
Qy 301 HLSKAVVDYEMKVQEANWELYNRQKAAEALLYEQEKQAEARRASADAAFFARQREAEAEL 360
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 301 HLSKAVVDYEMKVQEANWELYNRQKAAEALLYEQEKQAEARRASADAAFFARQREAEAEL 360
Qy 361 YAKQKEAEGLVAMGDAQSAYLSAMLGALGGSYAALRDYLMVSSGVYQEMARINADAIRGL 420
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 361 YAKQKEAEGLVAMGDAQSAYLSAMLGALGGSYAALRDYLMVSSGVYQEMARINADAIRGL 420
Qy 421 EPKISVWSNGAGAGGEVGEGGGAMKEVAGVYKMLPPLLTTVHEQTGMLPPAWMGTLTGGA 480
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 421 EPKISVWSNGAGAGGEVGEGGGAMKEVAGVYKMLPPLLTTVHEQTGMLPPAWMGTLTGGA 480
Qy 481 PSSTS 485
|||||
Db 481 PSSTS 485
Kawahara et al. also describes rice mRNA sequences having 100% sequence identity to instant SEQ ID NO: 7 (GenBank Accession No. AP014963) (sequence alignment is not shown) and 100% sequence identity to instant SEQ ID NO: 5 (GenBank Accession No. AP014966) (alignment shown below), as recited in claim 19. (Sequences obtained from GenBank can be accessed via https://www.ncbi.nlm.nih.gov/ website.)
RESULT 8
AP014966s2/c
LOCUS AP014966s2 11207287 bp DNA linear
COMMENT segment of length 11207287: from 12000001 to 23207287
DEFINITION Oryza sativa Japonica Group DNA, chromosome 10, cultivar:
Nipponbare, complete sequence.
ACCESSION AP014966
VERSION AP014966.1
DBLINK BioProject: PRJDB1747
BioSample: SAMD00000397
SOURCE Oryza sativa Japonica Group (Japanese rice)
ORGANISM Oryza sativa Japonica Group
Eukaryota; Viridiplantae; Streptophyta; Embryophyta; Tracheophyta;
Spermatophyta; Magnoliopsida; Liliopsida; Poales; Poaceae; BOP
clade; Oryzoideae; Oryzeae; Oryzinae; Oryza; Oryza sativa.
AUTHORS Kawahara,Y., de la Bastide,M., Hamilton,J.P., Kanamori,H.,
McCombie,W.R., Ouyang,S., Schwartz,D.C., Tanaka,T., Wu,J., Zhou,S.,
Childs,K.L., Davidson,R.M., Lin,H., Quesada-Ocampo,L.,
Vaillancourt,B., Sakai,H., Lee,S.S., Kim,J., Numa,H., Itoh,T.,
Buell,C.R. and Matsumoto,T.
TITLE Improvement of the Oryza sativa Nipponbare reference genome using
next generation sequence and optical map data
JOURNAL Rice (N Y) 6 (1), 4 (2013)
PUBMED 24280374
FEATURES Location/Qualifiers
source 1..23207287
/organism="Oryza sativa Japonica Group"
/mol_type="genomic DNA"
/cultivar="Nipponbare"
/db_xref="taxon:39947"
/chromosome="10"
assembly_gap 1..1000
/estimated_length=1000
/gap_type="telomere"
gene <44903..45178
/gene="Os10g0100200"
/locus_tag="OSNPB_100100200"
CDS <44903..45178
/gene="Os10g0100200"
/locus_tag="OSNPB_100100200"
/note="Os10t0100200-01: Hypothetical protein.;
start codon is not identified."
/codon_start=1
/protein_id="BAT09554.1"
/translation="GARGGALLWQVKTVPAPPLAAACHSIILWPPCRRSRRPPLATAR
RLRSPQPLPRLRPPRHSLQPVTSQPDDGVEVGEGRQGGVKGVVALAR"
gene complement(46217..58258)
/gene="Os10g0100300"
/locus_tag="OSNPB_100100300"
CDS complement(join(46217..46271,46356..46553,46681..46866,
55111..55202,55774..56016,56899..56971,57152..57233,
57366..57548,57631..57670,57782..57854,57990..58258))
/gene="Os10g0100300"
/locus_tag="OSNPB_100100300"
/note="Os10t0100300-01: NAD(P)-binding domain containing
protein."
/codon_start=1
/protein_id="BAT09555.1"
/translation="MEQAAKATISLSPPSYAGCCMAACPYRSTRHLRRGGGCSARSIS
SLRHAPSARVYAAAAAAATPESKSTKENDLVFIAGATGKVGSRAVREFIKLGFRVRAG
VRSAQRASSLVQSVEQLKVDDDATSPAERLEIVECDLEKQAQSDIVSAIGNAAIVVCS
IGASEKDILDVTGPYRIDYMATNNLVQAATAAKVEHFILVTSLGTNRIGFPAFLLNLF
WGVLCWKRRAEEALIGSGLPYTIVRPGGMERPTDAFKETHNLVVAVEDTYVGGLVSNL
QVAELIACIASNRRTAYCKVVEAIA ETTAPLLPTEDQLANIPSKRQPPPEPEVVQQGE
TPPKPIQQSQRPLSPYTAFVDLKPPSSPSPCPPSAAAPAPTSTDTAAAGSSSTLNSSA
TGTPISVDQPKQQQRPLSPYTRYEELKPPSSPSPTPPSAASSASVSASPDTPPAAAAS
SAALDSSANGTPITGDQLNQQQSPLSPYTRYEELKPPSSPTPSTPKL"
gene 67126..72969
/gene="Os10g0100500"
/locus_tag="OSNPB_100100500"
CDS join(67126..67396,67556..67674,67787..67896,69945..70383,
70479..70601,70773..71318,71697..72366,72480..72637)
/gene="Os10g0100500"
/locus_tag="OSNPB_100100500"
/note="Os10t0100500-01: Serine/threonine protein
kinase-related domain containing protein."
/codon_start=1
/protein_id="BAT09556.1"
/translation="MEILSPSPPPSPGPASASASASGSAMDSFIHRGAGWHFPRRDNV
DARVHVAVGRSPEKTLGLLRWAFRRFACAQVVLVHVHQPSPLIPTLLGKIPAAQATEE
LVLSHRKSEKDEMNKILLTYLTFCHRAQVQASLLVTENEQIHDGIITLVKDHGITKLV
MGSTPDTCFKLKASYGKASFMARNAPSFCEIWFVWRGRHIWTREAAAAIGNNISVYNE
DDVMIRKRIRFSSTSNNAESILDEGYISYEAQTPADRYEITISDNGQPNDYESLVDAN
HFCNIIVPNLQNAQSAFNSTFQPGSSVDMESLVLYPQEILDKNFKQVILEAERSRKDA
FVELLKRKDTESRVAGVIARAKASEFAQKQEMKMREELEALLTATKKQHEDLAENKEK
ATEGLDSSMRKLAILDARAKSIAFRMNEAVAELKLIQSSIGTLNQEIPKREKLELVHT
DQVERCAYNHIMLPNCSSTVCADDLYNFRELTLSDIKAATCKFSDSLKVLPRGLGCVY
KGEIMNRSVMIYKLHSCIIQSSMQFQQEVHLISKVRHPHLVTLIGACPDALCLVYEYV
PNGSLHDRLWSKCGIPQLPWKIRARIVAEISSALFFLHSCKPQMIVHGDLKLENILLD
ANLHCKIADCGISQLFMEDAKDADPEYRRSKPLTPKSDIYSFGIVILQLLTGKQAAGL
PSEVRRAMSSGKLWSLLDPTAGEWPLEVARRLAELGLKCSEAASPELLTPETVRDLEQ
LHLMRDNRQVPSFFLCPILKEVMHDPQVGADGLTYEGRAISELMDNGPPITPNHALRF
AIHDWLSQRSTPF"
CDS join(<71175..71318,71697..72366,72480..72637)
/gene="Os10g0100500"
/locus_tag="OSNPB_100100500"
/note="Os10t0100500-02: Similar to predicted protein.;
start codon is not identified."
/codon_start=1
/protein_id="BAT09557.1"
/translation="ATCKFSDSLKVLPRGLGCVYKGEIMNRSVMIYKLHSCIIQSSMQ
FQQEVHLISKVRHPHLVTLIGACPDALCLVYEYVPNGSLHDRLWSKCGIPQLPWKIRA
RIVAEISSALFFLHSCKPQMIVHGDLKLENILLDANLHCKIADCGISQLFMEDAKDAD
PEYRRSKPLTPKSDIYSFGIVILQLLTGKQAAGLPSEVRRAMSSGKLWSLLDPTAGEW
PLEVARRLAELGLKCSEAASPELLTPETVRDLEQLHLMRDNRQVPSFFLCPILKEVMH
DPQVGADGLTYEGRAISELMDNGPPITPNHALRFAIHDWLSQRSTPF"
3'UTR 72638..72969
/gene="Os10g0100500"
/locus_tag="OSNPB_100100500"
/note="Os10t0100500-01"
3'UTR 72638..72765
/gene="Os10g0100500"
/locus_tag="OSNPB_100100500"
/note="Os10t0100500-02"
gene complement(81232..82173)
/gene="Os10g0100700"
/locus_tag="OSNPB_100100700"
CDS complement(81232..82173)
/gene="Os10g0100700"
/locus_tag="OSNPB_100100700"
/note="Os10t0100700-01: Vitamin B6 biosynthesis protein
family protein."
/codon_start=1
/protein_id="BAT09558.1"
/translation="MASDGTDVVALYGGANGLSHKSGSFSVKVGLAQMLRGGVIMDVV
TPEQARIAEEAGACAVMALERVPADIRAQGGVARMSDPGLIRDIKRSVTIPVMAKARI
GHLVEAQILEAIGVDYVDESEVLTLADDAHHINKNNFRVPFVCGCRDLGEALRRIREG
AAMIRTKGEAGTGNVVEAVRHVRSVMGDIRALRSMDDDEVFSYAKRIAAPYDLVMQTK
QLGRLPVVQFAAGGVATPADAALMMQLGCDGVFVGSGIFKSGDPALRARAIVQAVTHY
SDPKILAEVSSGLGEAMVGINLSDPKIHVERFAARSD"
gene complement(94844..95488)
/gene="Os10g0100900"
/locus_tag="OSNPB_100100900"
CDS complement(94844..95488)
/gene="Os10g0100900"
/locus_tag="OSNPB_100100900"
/note="Os10t0100900-01: Hypothetical protein."
/codon_start=1
/protein_id="BAT09559.1"
/translation="MLVVVVVPLLMLGMACDAISVSVGSPKLDQAPWSRLPESFRDKG
ALVELKLRDQRVVLLPLSLHDSFPAASMESTELASPSTAEWSSLWAYHMTTGLDSLAL
ALTQTANQNLAGEEGEGRNPAAKSPLLETRVRLLSAGLQLVSLAEKVRICWAWAWAAA
RAGRRRRRRTAAAMVVLAGLGKGIGFLAFGFGIEPTTLLTSSLVNNTVANLPIM"
gene 94968..97409
/gene="Os10g0101000"
/locus_tag="OSNPB_100101000"
CDS 94968..97409
/gene="Os10g0101000"
/locus_tag="OSNPB_100101000"
/note="Os10t0101000-01: Serine/threonine protein kinase
domain containing protein."
/codon_start=1
/protein_id="BAT09560.1"
/translation="MAAAVLLLLLLPALAAAQAQAQQMRTFSANDTNWSPAESNRTLV
SNNGDFAAGFRPSPSSPAKFWFAVWVSANANESRPVVIWYAHNDDHSAVEGDANSVLS
IDAAGKLSWSDNGNSTTLWSRNFNSTSAPLSLNDSGSLDHGAWSSFGEPTDTLMASQA
IPSISNGTTTTTSITLQSQNGRFQLFNALTLQHGSSAYANITGNTALRNLTADGTLQL
AGGNPSQLIASDQGSTRRLRRLTLDDDGNLRLYSLQSKKGQWRVVWQLVQELCTIRGA
CQGEANICVPQGADNTTCVCPPGYRPQGLGCAPKLNYSGKGNDDKFVRMDFVSFSGGA
DTGVSVPGKYMTSLTPQNLADCQSKCRANASCVAFGYKLGGDRTCLHYTRLVDGYWSP
ATEMSTYLRVVESNNDPNNFTGMTTMIDTVCPVRLALPVPPKQGRTTIRNIAIITALF
AVELLAGVLSFWAFLRKYSQYREMARTLGLEYLPAGGPRRFSYAELKAATKEFSDLVG
RGAYGKVYRGELPDRRAVAVKQLDGVGGGEAEFWAEVTIIARMHHLNLVRMWGFCADK
EQRMLVYEYVPNGSLDKYLFAPGTGTQGDEEESNKRPLLDLHTRYRIALGVARAIA YL
HEECLEWVLHCDIKPENILLEDDFCPKVSDFGLSKLTSKKEKVTMSRIRGTRGYMAPE
WVIHREPITAKADVYSFGMVLLEIVSGRRNYGFRQDSVGSEDWYFPKWAFEKVYVERR
IDDIIDPRIVQAEAYDDDPASLATVERMVKTAMWCLQDRADMRPSMGKVAKMLEGTVE
ITEPVKPTIFCVQDD"
gene complement(100835..103317)
/gene="Os10g0101100"
/locus_tag="OSNPB_100101100"
CDS complement(join(100835..100918,101226..101271,
101377..101609,101733..101804,102103..102198,
102298..102381,102474..102575,102652..103317))
/gene="Os10g0101100"
/locus_tag="OSNPB_100101100"
/note="Os10t0101100-01: Peptidase S10, serine
carboxypeptidase family protein."
/codon_start=1
/protein_id="BAT09561.1"
/translation="MHLCTEAAVTRRRLLLLLLLLVTCSCLSARERSNSSSSSSRRVV
RHLPGFDGALPFELETGYVEVDRIAGVRLFYYFIRSESSPADDPLLLWLTGGPGCSAF
SGLVYEVGPLTFDVHGHGHGQLPRLLYKPESWTKRTNVIFLDSPVGTGFSYADTDAGF
RTGDTIAVHHILVFLNNWFQEVHPDFLSNPLYIAGDSYSGMIVPAVTFGIATSSPKPS
LNLKGYLLGNPVTDHNFDAPSKIPFAHGMGLISDQLYQAYKKSCSVKHNTQQQSVQCT
NTLDAIDECVKDIYGNHILEPYCTFASPHNPRIDKPFTSGTAEYTMSRIWANNDTVRE
Query Match 100.0%; Score 23; Length 11207287; Best Local Similarity 100.0%;
Matches 23; Conservative 0; Mismatches 0; Indels 0; Gaps 0;
Qy 1 GTGGGAGAGCTTGTCGTGCGGGG 23
|||||||||||||||||||||||
Db 6169404 GTGGGAGAGCTTGTCGTGCGGGG 6169382
Before the effective filing date of the invention, it would have been obvious to an ordinarily skilled artisan to inhibit or reduce expression level of the Importin α4 and flotillin1 (as described by Ma et al. in view of Cao et al. and DeBlasio et al.; see above) genes in a rice, as taught by Kawahara et al., with a realistic goal to obtain a rice plants capable to resist viral pathogen(s).
Before the effective filing date, an ordinarily skilled artisan would have been motivated to inhibit or reduce expression level of the Importin α4 and flotillin1 genes in a rice with a realistic goal to obtain a rice plants capable to resist viral pathogen(s).
Regarding claims 15-18 and 20-21; Traditional breeding methods based on random mutations induced by agents such as ethyl methanesulfonate (EMS) (as recited in claim 15), are long well-known in plant breeding1. Reducing or inhibiting expression of any gene(by deleting the gene itself or deleting the promoter sequence) in plant including in rice by CRISPR-Cas gene editing technique (as recited in claim 15) is also a routine and standard method and well-known before the effective filing date of the invention1. CRISPR-Cas can introduce substitution, deletion, addition/insertion of any number of nucleotides including 1, 2, or 3 nucleotides (as recited in claim 16), site specific mutagenesis, and gene knockouts in different alleles of any gene including Flotillin1 and Importin α4 genes (unless evidence to the contrary) in plants including rice, as recited in claim 15-17. Constructing a vector having sgRNA(s) and use of sgRNAs (as recited in claims 18 and 20-21) in CRISPR-Cas based genome editing for editing specific target gene(s) is also well-known and a standard practice in the art1.
Response to Applicant’s Argument
The Applicant’s response filed on 2/11/2026 is fully considered but not found persuasive.
The applicant argues that “Flot1… is required for seedling development” (page 2, para 1, line 3-4; page 2, para 3, line 2-3) implying that reduced Flot1 would prevent obtaining a transgenic seedling. The Applicant continues to argue, “Ma suggests that reducing Importin α4 expression levels lowers pathogen infectivity, Cao provides teaching contrary to the technical solution of the present application, proposing that reducing Flotillin1 expression levels diminishes plant resistance to pathogens while maintaining normal growth capacity and fertility...” (page 4, para 3, line 1-4).
The Applicant also argues about “unexpected effects” (page 4, para 5) in the invention while asserting that, “the claimed method is significantly different from the method of Ma, Cao, and Deblasio, and Ma, Cao, and Deblasio do not provide any teachings or suggestions which would motivate a skilled person in the art to readily envisage or obtain the claimed method” (page 5, para 1).
The Examiner disagrees. Cao describes that Flotillin1 (Flot1) is involved in endocytosis (page 2, para 1, line 12-13; page 2, last para, last 2 lines) which is long known to play a crucial part in diverse biological processes including response to pathogen attack. It would have been obvious that downregulation of Flot1 would reduce pathogen endocytosis, preventing the pathogenic bacteria from entering the host cell and establishing infection, and, thus increase, plant resistance to the said pathogen.
It is the opinion of the Applicant that, “reducing Flotillin1 expression levels diminishes plant resistance to pathogens”. The opinion of the Applicant is not supported by any evidence (MPEP 2145(I)).
Besides the teachings of Cao et al., DeBlasio et al. describes how Flot1 protein is involved (not in bacterial but) in viral pathogen interaction with the host plant, indicating that reduced expression of Flotillin1 has a realistic possibility to reduce endocytosis (as taught by Cao et al.) and prevents the viral pathogen(s) from entering the cell and establishing infection and, thus, increasing plant resistance against the virus(es).
Moreover, transgenic Arabidopsis lines with reduced Flot1 expression (reduction in Flot1 transcript levels) via microRNA based downregulation were able to develop seedlings, but with a reduction in shoot and root meristem size plus retardation in seedling growth (Li et al., A Membrane Microdomain-Associated Protein, Arabidopsis Flot1, Is Involved in a Clathrin-Independent Endocytic Pathway and Is Required for Seedling development, 2012, The Plant Cell, 24:2105–2122; abstract).
It is prudent to mention here that Applicant’s argument of “unexpected effects” resulting from simultaneous reduction of both Flotillin1 and Importin α4 expression levels in plants is not commensurate in scope with the claims (MPEP 716.02(d)). The claims comprise any pathogen while the example provided by the Applicant mention only one viral pathogen (RSV NP) (spec, page 5, Table 1).
Conclusion
No claim is allowed.
THIS ACTION IS MADE FINAL. Applicant is reminded of the extension of time policy as set forth in 37 CFR 1.136(a).
A shortened statutory period for reply to this final action is set to expire THREE MONTHS from the mailing date of this action. In the event a first reply is filed within TWO MONTHS of the mailing date of this final action and the advisory action is not mailed until after the end of the THREE-MONTH shortened statutory period, then the shortened statutory period will expire on the date the advisory action is mailed, and any nonprovisional extension fee (37 CFR 1.17(a)) pursuant to 37 CFR 1.136(a) will be calculated from the mailing date of the advisory action. In no event, however, will the statutory period for reply expire later than SIX MONTHS from the mailing date of this final action.
Communication
Any inquiry concerning this communication or earlier communications from the examiner should be directed to JAY CHATTERJEE whose telephone number is (703)756-1329. The examiner can normally be reached (Mon - Fri) 8.30 am to 5.30 pm..
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Jay Chatterjee
Patent Examiner
Art Unit 1662
/Jay Chatterjee/Examiner, Art Unit 1662
/BRATISLAV STANKOVIC/Supervisory Patent Examiner, Art Units 1661 & 1662
1Chen et al. (CRISPR/Cas Genome Editing and Precision Plant Breeding in Agriculture, 2019, Annu. Rev. Plant Biol. 2019. 70:667–97) provides the evidence that EMS mutagenesis (page 683, last para, line 4); CRISPR-Cas mediated genome editing using single or multiplex of sgRNA(s) (reads on sgRNa1, sgRNA2 etc.) (page 671, para 1, line 1-5; page 673, para 2, line 2; page 674, para 4, line 3) introducing substitution(s), indel(s) (deletion, addition/insertion) (page 679, Fig.5; page 671, para 2, line 13-14), site specific mutation(s), and gene knock-outs (page 678, para 2) in plants including rice (page 684, para 1, line 1-4) is a routine and standard practice in the art.