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 .
Election/Restrictions
Applicant’s election of Group I in the reply filed on 5/4/2026 is acknowledged.
Because applicant did not distinctly and specifically point out the supposed errors in the restriction requirement, the election has been treated as an election without traverse (MPEP § 818.01(a)).
The requirement is still deemed proper and is therefore made FINAL.
Claim Status
Claims 1-24 are pending.
Claims 16-24 are withdrawn from consideration as part of non-elected groups.
Claims 1-15 are being examined.
Information Disclosure Statement
Applicant’s IDS, submitted on 07/25/2024, has been considered only to the extent of the English translations submitted. A signed copy is attached.
Specification
The disclosure is objected to because of the following informalities:
Instant specification describes, “…. Korean Patent Publication No. 10-2017-0138657 discloses that when the DWF5 (delta 5, 7-sterol deta 7 reductase, DWARF5), CPD (constitutive photomorphogenic DWAF, DWF3) and SMT1 (sterol methyltransferase 1) genes are deleted, lettuce enriched with 7-dehydrocampesterol, a precursor of vitamin D, can be produced. However, no studies have been conducted on the production of tomatoes enriched with 7-dehydrocholesterol as a precursor of vitamin D3” (spec, p.1, para 5, last 6 lines). However, Korean Patent Publication No. 10-2017-0138657 (KR1020170138657) is not related to lettuce enriched with 7-dehydrocampesterol but with “Method and apparatus for battery charging”.
Appropriate correction is required.
The disclosure is objected to because it contains an embedded hyperlink and/or other form of browser-executable code. There is an embedded hyperlink and/or other form of browser-executable code in page 17 (para 4).
Applicant is required to delete the embedded hyperlink and/or other form of browser-executable code; references to websites should be limited to the top-level domain name without any prefix such as http:// or other browser-executable code. See MPEP § 608.01.
Claim Objections
Claim 1-15 are objected. Claim 1 recites, “A transformed tomato”, and it is apparent, however, that “tomato” in the claims is not just referring to the fruit, but to a tomato plant. It is suggested to amend claim 1 to insert the term “plant” in line 1 after “A transformed tomato”, and dependent claims should then also be amended accordingly to avoid lack of antecedent basis issues.
Claims 6, 10 and 13 are objected to because of the following informalities: Claims 6, 10 and 13 recites, “… the genetic engineering is induced by modification in the nucleic acid sequence….”. The modifications in the nucleic acid sequence of the genes do not induce any genetic engineering. Rather, it is the other way round. The modifications of the genes are achieved by the genetic engineering.
Appropriate correction is required.
Claim Rejections - 35 USC § 112(b)
The following is a quotation of 35 U.S.C. 112(b):
(b) CONCLUSION.—The specification shall conclude with one or more claims particularly pointing out and distinctly claiming the subject matter which the inventor or a joint inventor regards as the invention.
The following is a quotation of 35 U.S.C. 112 (pre-AIA ), second paragraph:
The specification shall conclude with one or more claims particularly pointing out and distinctly claiming the subject matter which the applicant regards as his invention.
Claims 1-2 and 4-15 are rejected under 35 U.S.C. 112(b) or 35 U.S.C. 112 (pre-AIA ), second paragraph, as being indefinite for failing to particularly point out and distinctly claim the subject matter which the inventor or a joint inventor (or for applications subject to pre-AIA 35 U.S.C. 112, the applicant), regards as the invention.
The term “high” in claims 1-2 is a relative term which renders the claim indefinite. The term “high” is not defined by the claim, the specification does not provide a standard for ascertaining the requisite degree or range, and one of ordinary skill in the art would not be reasonably apprised of the scope of the invention. Thus, it is not clear to the Examiner what concentration of 7-dehydrocholesterol would be considered “high” enough to satisfy the claim limitation.
All the claims depending from claim 1 or 2 inherit the indefiniteness of claims 1-2.
Claim Rejections - 35 USC § 102(a)(1)
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 the appropriate paragraphs of 35 U.S.C. 102 that form the basis for the rejections under this section made in this Office action:
A person shall be entitled to a patent unless –
(a)(1) the claimed invention was patented, described in a printed publication, or in public use, on sale, or otherwise available to the public before the effective filing date of the claimed invention.
(a)(2) the claimed invention was described in a patent issued under section 151, or in an application for patent published or deemed published under section 122(b), in which the patent or application, as the case may be, names another inventor and was effectively filed before the effective filing date of the claimed invention.
Claims 1-7 are rejected under 35 U.S.C. 102(a)(1) as being anticipated by Mizutani et al. (WO 2019/163601 Al).
Mizutani et al. teaches a transgenic (reads on “transformed”) tomato plants (p. 16, para 0049, line 13-14) with increased amount (reads on “high”) of 7-dehydrocholcsterol (7-DHC) (abstract; P.3, para 0006, line 1-2; p.7, para 0017). Formula of 7-dehydrocholcsterol is well known in the art and also taught by Mizutani et al. (Fig. 1-2). Figure 13 describes 7-DHC content in tomato (hairy) roots (as recited in claim 2) in which both the DWF5 genes, including the DWF5-1 gene, are modified (as recited in claim 6) by deleting (as recited in claim 7) or knocking the gene out (p. 5, para 0009, Fig. 13; Fig. 13), which reads on to genetically engineered tomato to reduce the expression or activity of the DWF5-1 gene or the DWF5-1 protein, as recited in claim 4.
The DWF5-1 protein set forth by instant SEQ ID NO: 2 (spec, p.5, para 3, line 4-5) is encoded by the DWF5-1 gene comprising the nucleic acid sequence set forth by SEQ ID NO: 63 (spec, p.6, para 6, line 1-2), as recited in claim 5. Mizutani et al. describes a DWF5 protein (SEQ ID NO: 1) having 100% sequence identity to instant SEQ ID NO: 2, as shown below.
Title: US-18-832-478-2
Perfect score: 2365
Sequence: 1 MVENKLVHSPLITYGSMLSL..........GKYWKLYCEKVPYRVIPGIY 434
Scoring table: BLOSUM62
Gapop 10.0 , Gapext 0.5
Searched: 1 seqs, 432 residues
Database : AASEQ2_05112026_122557.fasta:*
RESULT 1
AASEQ2_05112026_122557
Query Match 100.0%; Best Local Similarity 100.0%; Score 2365; Length 435;
Matches 434; Conservative 0; Mismatches 0; Indels 0; Gaps 0;
Qy 1 MVENKLVHSPLITYGSMLSLLSFTPPFVILMWYTNVHADGSILKTFNHLRENGLQGLIDI 60
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 1 MVENKLVHSPLITYGSMLSLLSFTPPFVILMWYTNVHADGSILKTFNHLRENGLQGLIDI 60
Qy 61 WPKPTAIA GKLIICYALFEAALQLLLPGKTVEGPISPTGHRPVYKANGMAAYAVTLITYI 120
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 61 WPKPTAIA GKLIICYALFEAALQLLLPGKTVEGPISPTGHRPVYKANGMAAYAVTLITYI 120
Qy 121 SLWWFGIFNPAIVYDHLGEIFSTLIFGSLVFCVLLYIKGHVAPSSTDSGSSGNIIVDFYW 180
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 121 SLWWFGIFNPAIVYDHLGEIFSTLIFGSLVFCVLLYIKGHVAPSSTDSGSSGNIIVDFYW 180
Qy 181 GMELYPRIGKHFDIKVFTNCRFGMMSWAVLAVTYCIKQHEEYGRVSDSMLVNTILMLVYV 240
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 181 GMELYPRIGKHFDIKVFTNCRFGMMSWAVLAVTYCIKQHEEYGRVSDSMLVNTILMLVYV 240
Qy 241 TKFFWWEAGYWNTMDIAHDRAGFYICWGCLVWVPSIYTSPGMYLVKQPVNLGLQLSLYIL 300
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 241 TKFFWWEAGYWNTMDIAHDRAGFYICWGCLVWVPSIYTSPGMYLVKQPVNLGLQLSLYIL 300
Qy 301 VAGLLCIYINYDCDRQRQEFRRTNGKCTVWGKTPSKIVAAYTTTSGEKKTSLLLTSGWWG 360
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 301 VAGLLCIYINYDCDRQRQEFRRTNGKCTVWGKTPSKIVAAYTTTSGEKKTSLLLTSGWWG 360
Qy 361 LARHFHYVPEILAAFFWSVPALFNHFIPYFYVIFLIILLLDRAKRDDDRCKAKYGKYWKL 420
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 361 LARHFHYVPEILAAFFWSVPALFNHFIPYFYVIFLIILLLDRAKRDDDRCKAKYGKYWKL 420
Qy 421 YCEKVPYRVIPGIY 434
||||||||||||||
Db 421 YCEKVPYRVIPGIY 434
In the wild type tomato plants, 7-DHC is present in amounts below the detection limit, but as shown in Figure 13, it was found that knocking out both the DWF5 genes leads to the accumulation of 7-DHC in amounts as high as 10 µg/gFW, i.e., 1 mg per 100 g of (fresh) weight (p. 36, para 0110, last 3 lines), as recited in claim 3. Instant description does not define or describe “weight”- whether that is dry weight or fresh weight. The Examiner interprets that as fresh weight because the Applicant describes “certain amount of each sample was weighed” (i.e., fresh weight) to analyze vitamin D3 precursor (spec, p.25, Example 5.1., para 6, line 1).
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.
The factual inquiries for establishing a background for determining obviousness under 35 U.S.C. 103 are summarized as follows:
1. Determining the scope and contents of the prior art.
2. Ascertaining the differences between the prior art and the claims at issue.
3. Resolving the level of ordinary skill in the pertinent art.
4. Considering objective evidence present in the application indicating obviousness or nonobviousness.
Claims 8-10 are rejected under 35 U.S.C. 103 as being unpatentable over by Mizutani et al. as applied to claims 1-7 above, and further in view of Nomura et al. (Accumulation of 6-deoxocathasterone and 6-deoxocastasterone in Arabidopsis, pea and tomato is suggestive of common rate-limiting steps in brassinosteroid biosynthesis, 2001, Phytochemistry, 57:171-178) and Bajguz et al. (Comprehensive Overview of the Brassinosteroid Biosynthesis Pathways: Substrates, Products, Inhibitors, and Connections, 2020, Front. Plant Sci. 11:1034) and He et al. (Cloning of gene CYP90A1-like in Tomato, 2016, Directly submitted to GenBank on 2016).
Claim 8 depends from claim 4 and is drawn to a tomato plant engineered to reduce the expression or activity of the CPD gene or CPD protein, besides having reduced expression and activity of DWF5-1 gene.
Mizutani et al. describes a tomato plant engineered to reduce the expression or activity of the DWF5-1 protein encoded by the DWF5-1 gene comprising the sequence set forth by instant SEQ ID NO: 63, as discussed above. Mizutani et al. also describes that cycloartenol is converted to 7-dehydrocholesterol by several enzymes, and then 7-dehydrocholesterol is converted to cholesterol by the enzyme encoded by the protein DWF5-1 (Figure 1).
However, Mizutani et al. does not describe reducing the expression or activity of the CPD gene or CPD protein in the tomato plant.
Nomura et al. describes the conversions to 6-deoxoteasterone, castasterone, and 6-deoxocathasteonre are important rate-limiting steps in brassinosteroid (BR) biosynthesis in plants (abstract). The most abundant BR is 6-deoxocastasterone closely followed by the second most abundant BR 6-deoxocathasterone in all the plant tissues examined (p.175, left column, para 2, line 1-3). The C-23 hydroxylation of 6-deoxocathasterone to 6-deoxoteasterone appears also to be a rate-limiting step. In Arabidopsis, C-23 hydroxylation is catalyzed by a CPD protein, also named CYP90A. The transcription of the CPD gene has been found to be suppressed by active BRs through a feedback mechanism indicating that the synthesis of 6-deoxoteasterone is also an important control point in BR biosynthesis (p.5, left column, para 2, last 6 lines).
Bajguz et al. describes that sterols including 7-dehydrocholesterol and cycloartenol are the precursors of all the BR biosynthesis (Fig. 1).
It would have been obvious to an ordinarily skilled artisan to modify or mutate the CPD gene using any routine and standard genetic engineering or genome editing technique (as recited in claim 10) so that the CPD (CYP90A) protein, as described by Nomura et al., is inactivated so that BR biosynthesis is severely affected. That (i.e., the lack of the crucial CPD protein) would reduce or abolish the conversions of sterols including cycloartenol and 7-dehydrocholesterol to BRs in the transformed tomato plants comprising DWF5 genes knocked, as described by Mizutani et al.
An ordinarily skilled artisan would have been motivated to modify and mutate/deactivate the CPD protein and/or knock out the CPD gene with a realistic expectation to increase sterol including 7-dehydrocholesterol concentration further in a commercially important tomato plant that already has its DWF5 genes knocked-out
Regarding claim 9, He et al. (Cloning of gene CYP90A1-like in Tomato, 2016, Directly submitted to GenBank on 2016) describes a tomato CPD/CYP90A gene which encodes a protein having a 100% sequence identity to instant SEQ ID NO: 5, as shown below.
Title: US-18-832-478-5
Perfect score: 1742
Sequence: 1 gcagagcgttttgcatacaa..........taaagatttaattgccctca 1742
Searched: 1 seqs, 1749 residues
Database : NASEQ2_05282026_171818.fasta:*
RESULT 1
NASEQ2_05282026_171818
Best Local Similarity 100.0%; Query Match 99.4%; Score 1731; DB 1; Length 1749;
Matches 1731; Conservative 0; Mismatches 0; Indels 0; Gaps 0;
Qy 3 AGAGCGTTTTGCATACAAAAAAAAATTAATTAACATGGATACCATCGATCTTTTTCTCTA 62
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 2 AGAGCGTTTTGCATACAAAAAAAAATTAATTAACATGGATACCATCGATCTTTTTCTCTA 61
Qy 63 TCTTTTTCTCTCTGTTTTCACCTTTTATCTCCTCCGGTGTATGGCGGCGGCCCATCTTCG 122
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 62 TCTTTTTCTCTCTGTTTTCACCTTTTATCTCCTCCGGTGTATGGCGGCGGCCCATCTTCG 121
Qy 123 CGGGCGTAAAACTCGACTCCCGCCGGGAACCCTTGGCCTCCCGTTTATCGGAGAAACCCT 182
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 122 CGGGCGTAAAACTCGACTCCCGCCGGGAACCCTTGGCCTCCCGTTTATCGGAGAAACCCT 181
Qy 183 CCAGTTAATTTCTGCGTACAAAACGGAAAATCCTGAACCGTTCATCGATGACCGTGTTTC 242
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 182 CCAGTTAATTTCTGCGTACAAAACGGAAAATCCTGAACCGTTCATCGATGACCGTGTTTC 241
Qy 243 AAAATACGGCAGCATTTTCACGACCCATGTTTTTGGTGAACCGACGGTTTTCTCTGCTGA 302
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 242 AAAATACGGCAGCATTTTCACGACCCATGTTTTTGGTGAACCGACGGTTTTCTCTGCTGA 301
Qy 303 CCCGGAAACGAACCGGTTTATTTTGCAGAATGAAGGTCGGCTTTTTGAGTCGAGTTATCC 362
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 302 CCCGGAAACGAACCGGTTTATTTTGCAGAATGAAGGTCGGCTTTTTGAGTCGAGTTATCC 361
Qy 363 CGGTTCGATACAGAATTTACTTGGGAGATACTCTCTGTTGCTTATGAGAGGAAGTCTACA 422
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 362 CGGTTCGATACAGAATTTACTTGGGAGATACTCTCTGTTGCTTATGAGAGGAAGTCTACA 421
Qy 423 CAAACGAATGCATTCATTAACTATGAGTTTTGCTAATTCTTCCATTCTTAAAGATCATCT 482
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 422 CAAACGAATGCATTCATTAACTATGAGTTTTGCTAATTCTTCCATTCTTAAAGATCATCT 481
Qy 483 GTTGGGCGATATAGATCGATTGGTTAGGCTTAATTTGGATTCATGGACCGGCAGGGTTTT 542
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 482 GTTGGGCGATATAGATCGATTGGTTAGGCTTAATTTGGATTCATGGACCGGCAGGGTTTT 541
Qy 543 CCTCATGGACGAGGCTAAGAAGATAACGTTTAATCTAACAGTGAAGCAGTTGATGAGTTT 602
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 542 CCTCATGGACGAGGCTAAGAAGATAACGTTTAATCTAACAGTGAAGCAGTTGATGAGTTT 601
Qy 603 TGATCCATGTGAGTGGACAGAGAATCTGATGAAAGAGTATATGCTTGTTATTGAAGGTTT 662
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 602 TGATCCATGTGAGTGGACAGAGAATCTGATGAAAGAGTATATGCTTGTTATTGAAGGTTT 661
Qy 663 CTTCTGCATTCCTTTGCCTATTTTCTCATCCACCTATCGCAAGGCCATTCAAGCGAGAAC 722
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 662 CTTCTGCATTCCTTTGCCTATTTTCTCATCCACCTATCGCAAGGCCATTCAAGCGAGAAC 721
Qy 723 GAAAGTAGCGGAGGCGTTGGGATTGGTAGTGAGAGATCGGAGAAAAGAACGAGACGGAGG 782
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 722 GAAAGTAGCGGAGGCGTTGGGATTGGTAGTGAGAGATCGGAGAAAAGAACGAGACGGAGG 781
Qy 783 AGAACGAAAGAATGATATGTTGGAGGCATTGTTCGAAGGAGACGGAGTTGAAGGAGTAGG 842
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 782 AGAACGAAAGAATGATATGTTGGAGGCATTGTTCGAAGGAGACGGAGTTGAAGGAGTAGG 841
Qy 843 ATTTTCCGATGAGGAAATTGTTGATTTTATACTGGCGTTGCTTGTTGCTGGCTATGAAAC 902
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 842 ATTTTCCGATGAGGAAATTGTTGATTTTATACTGGCGTTGCTTGTTGCTGGCTATGAAAC 901
Qy 903 TACCTCCACCATCATGACCCTTGCTGTAAAATTCCTCACGGAGACACCCCGTGCTCTATC 962
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 902 TACCTCCACCATCATGACCCTTGCTGTAAAATTCCTCACGGAGACACCCCGTGCTCTATC 961
Qy 963 ACTACTTAAGGAAGAGCACGAGGAGATCAGGTTAAGAAAAGGTGAAGTTAAGTCTTTACA 1022
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 962 ACTACTTAAGGAAGAGCACGAGGAGATCAGGTTAAGAAAAGGTGAAGTTAAGTCTTTACA 1021
Qy 1023 ATGGGAAGATTACAAGTCAATGCCCTTCACTCAATGCGTTGTTAATGAAACTCTGCGAAT 1082
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 1022 ATGGGAAGATTACAAGTCAATGCCCTTCACTCAATGCGTTGTTAATGAAACTCTGCGAAT 1081
Qy 1083 CGCTAACATAATTAGTGGAGTGTTCAGGAGAGCCATGACTGACATAAATATCAAAGGTTA 1142
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 1082 CGCTAACATAATTAGTGGAGTGTTCAGGAGAGCCATGACTGACATAAATATCAAAGGTTA 1141
Qy 1143 TACCATTCCAAAAGGTTGGAAGGTTTTTGCTTCTCTCCGAGCTGTTCATCTAGACCATGA 1202
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 1142 TACCATTCCAAAAGGTTGGAAGGTTTTTGCTTCTCTCCGAGCTGTTCATCTAGACCATGA 1201
Qy 1203 ACATTTCAAAGATGCGCGTACATTTGATCCATGGAGGTGGCAGAGTAGTGCAGGACCGAC 1262
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 1202 ACATTTCAAAGATGCGCGTACATTTGATCCATGGAGGTGGCAGAGTAGTGCAGGACCGAC 1261
Qy 1263 AAGCTCACCTAACGTATTCACACCATTTGGTGGTGGACCTCGTCGATGTCCTGGTTACGA 1322
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 1262 AAGCTCACCTAACGTATTCACACCATTTGGTGGTGGACCTCGTCGATGTCCTGGTTACGA 1321
Qy 1323 GCTTGCTAGAGTGGAACTCTCTGTTTTCCTTCACCACTTGGTGACTCGTTTTAGTTGGGT 1382
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 1322 GCTTGCTAGAGTGGAACTCTCTGTTTTCCTTCACCACTTGGTGACTCGTTTTAGTTGGGT 1381
Qy 1383 TCCGGCTGAGGCAGATAAGTTAGTTTTCTTCCCAACGACAAGGATGCTGAAGCGATATCC 1442
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 1382 TCCGGCTGAGGCAGATAAGTTAGTTTTCTTCCCAACGACAAGGATGCTGAAGCGATATCC 1441
Qy 1443 AATTAACGTCCAACATCGAAGCTTGTTTGAGCAGAAAGAAGAAGAGAAGGGTTCGTGAGA 1502
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 1442 AATTAACGTCCAACATCGAAGCTTGTTTGAGCAGAAAGAAGAAGAGAAGGGTTCGTGAGA 1501
Qy 1503 TTGTAGAATGCAGAGTGGAGGGAGGTGTATGAAAGGTGTTAAATAAGACAATAGTATATA 1562
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 1502 TTGTAGAATGCAGAGTGGAGGGAGGTGTATGAAAGGTGTTAAATAAGACAATAGTATATA 1561
Qy 1563 GGTGTTAGCATATACTTAGAAAATTAAATTATTTGTAGTGAATTGAGCAGAGATTAACAT 1622
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 1562 GGTGTTAGCATATACTTAGAAAATTAAATTATTTGTAGTGAATTGAGCAGAGATTAACAT 1621
Qy 1623 TCCTCTGATCTGCAAGGGGAATCAGGAAGATTTTATTCTGAGCAATTGGTTATTATATAA 1682
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 1622 TCCTCTGATCTGCAAGGGGAATCAGGAAGATTTTATTCTGAGCAATTGGTTATTATATAA 1681
Qy 1683 TAGTAATGCACAGTTTTTAATGTATGTTAATTAGCATGATTAAAGATTTAA 1733
|||||||||||||||||||||||||||||||||||||||||||||||||||
Db 1682 TAGTAATGCACAGTTTTTAATGTATGTTAATTAGCATGATTAAAGATTTAA 1732
Claims 11-15 are rejected under 35 U.S.C. 103 as being unpatentable over by Mizutani et al. in view of Nomura et al., Bajguz et al. and He et al. as applied to claims 8-10 above, and further in view of Chen et al. (Cholesterol accumulation by suppression of SMT1 leads to dwarfism and improved drought tolerance in herbaceous plants, 2018, Plant Cell Environ., 41:1417-1426).
Claim 11 depends on claim 8 and is drawn to a reduced expression or activity of the SMT1 gene or SMT1 protein in the tomato plant of claim 8.
Mizutani et al. in view of Nomura et al. and Bajguz et al. describe a transgenic or genome edited tomato plant with knocked out DWF5 genes including DWF5-1 gene and deactivated/knocked out CPD gene, as discussed above. Mizutani et al. also describes that SMT1 enzyme converts cycloartenol to 24-methylene cycloartenol (Fig. 1) which subsequently converted to BRs via various intermediated and enzymes, as described by Bajguz et al. (Fig. 1). The same pool of cycloartenol is also converted to cycloartanol (by DWF1H) enzyme which ultimately produces 7-dehydrocholesterol (Mizutani et al., Fig. 1).
However, Mizutani et al. in view of Nomura et al., Bajguz et al. and He et al. do not describe modifying SMT1 gene expression or SMT1 protein activity.
Chen et al. describes that mutating and inactivating the SMT1 (Sterol Methyltransferase 1) gene with significantly reduced transcript level enhanced drought tolerance and increased cholesterol accumulation in bermudagrass (abstract). Chen et al. also describes that knock-down of OsSMT1 expression resulted in similar phenotypic changes in transgenic rice (abstract).
Before the effective filing date of the invention, it would have been obvious to an ordinarily skilled artisan to modify/mutate the homologue of the SMT1 genes of bermudagrass (CdSMT1) and/or rice (OsSMT1), as described by Chen et al., in tomato. A tomato gene (Gene ID: Solyc01g087560.3) encoding a tomato protein comprising the highest sequence identity to CdSMT1 and OsSMT1 is identified. The protein set forth by gene ID Solyc01g087560.3 comprises 100% sequence identity to instant SEQ ID NO: 8, which is encoded by the polynucleotide sequence set forth by SEQ ID NO: 7 (spec, p.10, para 8), as recited in claim 12, as shown below.
Title: US-18-832-478-8
Perfect score: 1747
Sequence: 1 MSKQGAFDLAFGVGGRIGKD..........SQRVQAFLEKAAEGLVGGAK 326
Searched: 1 seqs, 346 residues
Database : AASEQ2_05122026_171626.fasta:*
RESULT 1
AASEQ2_05122026_171626
Best Local Similarity 100.0%; Query Match 100.0%; Score 1747; DB 1; Length 346;
Matches 326; Conservative 0; Mismatches 0; Indels 0; Gaps 0;
Qy 1 MSKQGAFDLAFGVGGRIGKDEVLSAVDKYEKYHGYYGGEEDERKNNYTDMVNKYYDLCTS 60
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 1 MSKQGAFDLAFGVGGRIGKDEVLSAVDKYEKYHGYYGGEEDERKNNYTDMVNKYYDLCTS 60
Qy 61 FYEYGWGESFHFAPRWKEESLQESIKRHEHFLALQLGLKPGQKVLDVGCGIGGPLREIAR 120
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 61 FYEYGWGESFHFAPRWKEESLQESIKRHEHFLALQLGLKPGQKVLDVGCGIGGPLREIAR 120
Qy 121 FSSTSVTGLNNNEYQISRGQVLNRKVGLDQTCNFVKGDFMKMPFPDNSFDAVYAIEATCH 180
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 121 FSSTSVTGLNNNEYQISRGQVLNRKVGLDQTCNFVKGDFMKMPFPDNSFDAVYAIEATCH 180
Qy 181 APDPVGCYREIYRVLKPGQCFAVYEWCMTDAYNPNNEEQKRIKEEIELGNGLPEIRSTQQ 240
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 181 APDPVGCYREIYRVLKPGQCFAVYEWCMTDAYNPNNEEQKRIKEEIELGNGLPEIRSTQQ 240
Qy 241 CLEAARKAGFEVVWDKDLAEDSPVSWYMPLDTSHFSLSSFRLTAVGRLFTRNLVSALEYV 300
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 241 CLEAARKAGFEVVWDKDLAEDSPVSWYMPLDTSHFSLSSFRLTAVGRLFTRNLVSALEYV 300
Qy 301 GVAPKGSQRVQAFLEKAAEGLVGGAK 326
||||||||||||||||||||||||||
Db 301 GVAPKGSQRVQAFLEKAAEGLVGGAK 326
Before the effective filing date of the invention, the ordinarily skilled artisan would have been motivated to modify/mutate including knocking out the tomato SMT1 gene (Gene ID: Solyc01g087560.3) (as recited in claims 11 and 13) (homolog of CdSMT1 and OsSMT1) in the tomato plant already having the mutated/knocked-out DWF5-1 and CPD genes, as described above, with a realistic goal to increase production of 7-dehydrocholesterol (as indicated by Mizutani et al.) and enhanced drought tolerance (as described by Chen et al).
Mizutani et al., describes that the same pool of cycloartenol is used as a substrate by two different enzymes. The first enzyme, DWF1H, produces cycloartanol which, in turn, produces 7-dehydrocholesterol (Mizutani et al., Fig. 1). The second enzyme, SMT1, converts cycloartenol to 24-methylene cycloartenol (Fig. 1) which subsequently is converted to BRs via various intermediates and enzymes, as described by Bajguz et al. (Fig. 1) and as discussed above. Thus, inactivating or knocking out the STM1 would have the realistic expectation to increase the conversion rate of DWF1H which ultimately is expected to increase 7-dehydrocholesterol concentration. Knocking out the CPD protein would also be expected to reduce or abolish production of BRs, and thus, increase the flux of 7-dehydrocholesterol production. However, cholesterol production is reduced/abolished because of knocking out the DWF5 genes, as described above. Mutation in the SMT1 gene is also expected to increase drought tolerance, an important agronomic trait, in the tomato plant.
Regarding claims 14-15, producing a homozygote plant and seeds thereof is a routine and standard practice in the art for maintenance of the trait and propagation of the tomato plant.
Conclusion
No claim is allowed.
Communication
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JC
/Jay Chatterjee/Examiner, Art Unit 1662
/BRATISLAV STANKOVIC/Supervisory Patent Examiner, Art Units 1661 & 1662
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