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 .
DETAILED ACTION
1. The Office acknowledges the receipt of Applicant’s Request for Continued Examination filed August 28, 2025. Claims 1, 3, 11, 13-15, 19, 23, 25-28, 31, 32, 34, 36 and 37 are pending. Claims 13-15, 19, 23, 26-28, 31, 32 and 34 are withdrawn. Claims 1, 3, 11, 25, 36 and 37, to the extent of SEQ ID NO:3, 11(d) and 25(d), are examined.
All previous rejections not set forth below have been withdrawn.
The text of those sections of Title 35, U.S. Code not included in this action can be found in a prior Office action.
Claim Objections
2. Claims 1, 3, 11, 25, 36 and 37 are objected to because of the following:
In claim 1, SEQ ID NO:1 should be deleted because it is an Arabidopsis sequence. SEQ ID Nos. 3, 5 and 7 are A, B and D genomes of wheat. The grouping is an improper grouping. Moreover, per Applicant’s claim amendments and Applicant’s Remarks filed August 28, 2025, Applicant stated that Arabidopsis is not a crop species.
In claim 1, the acronym “BLUS1” should be spelled out the first time it is used with the acronym in parentheses.
In claim 37, “one or more mutations” should be amended to “at least one mutation” for language consistency with claim 1.
Dependent claims are included.
Appropriate correction is required.
Claim Rejections - 35 USC § 112(b)
3. Claims 1, 3, 11, 25, 36 and 37 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 pre-AIA the Applicant regards as the invention.
In claim 1, it is unclear how “crop” is defined: a plant, more than one plant, plant(s) in a growth medium, a particular type of plant, etc. Page 10, lines 25-26 of the December 17, 2017 specification, defines a crop plant as “any plant which is grown on a commercial scale for human or animal consumption or use.” However, there is no definition given for “crop”. It is suggested “crop” be amended to “plant”. All subsequent recitations of “crop” are rejected.
In claim 1, it is unclear where the “introducing at least one mutation” occurs: in a plant? Where is the nucleic acid sequence encoding the BLUS1 polypeptide?
In claim 1, it is unclear whether a sequence having 80% sequence identity to SEQ ID NO:3 also encodes a BLUS1 polypeptide. For examination purpose, the Office interprets the 80% sequence identity to be limited to a BLUS1 polypeptide.
In claim 1, it is unclear what “a control or wild-type” refers to: a control or wild-type crop? For proper comparative basis, plant must be compared to plant, and crop must be compared to crop.
In claim 11, it is unclear whether the “introducing and/or identifying” step replaces the “reducing or abolishing” or the “introducing” step of claim 1. If so, claim 11 does not further limit claim 1. Claim 11 should further limit claim 1. As written, claim 11 does not require the “introducing” step.
In claim 36, soybean is not a cereal crop.
In claim 37, it is unclear what region of SEQ ID Nos. 1, 3, 5 and 7, or what region of a sequence having 80% sequence identity to SEQ ID Nos. 1, 3, 5 and 7, is the C-terminus and the N-terminus. Where do the C and N termini begin and end?
In claim 37, it is unclear whether the “BLUS1” of claim 37 is the same as the “BLUS1 polypeptide” of claim 1, or Applicant is referring to a BLUS1 gene. For examination purpose, the Office interprets “BLUS1” to be the “BLUS1 polypeptide” of claim 1. Otherwise, the “BLUS1” gene lacks antecedence.
Clarification and/or correction is required.
Claim Rejections - 35 USC § 112(a)
4. Claims 1-3, 8, 11, 25 and 36 are rejected under 35 U.S.C. 112(a) or 35 U.S.C. 112 (pre-AIA ), first paragraph, as failing to comply with the written description requirement. The claim(s) contains subject matter which was not described in the specification in such a way as to reasonably convey to one skilled in the relevant art that the inventor or a joint inventor, or for pre-AIA the inventor(s), at the time the application was filed, had possession of the claimed invention.
Applicant disclosure is as follows. In Example 1, the specification discloses that Arabidopsis blus1 mutants show reduced gs (stomatal conductance) when total red light illumination was replaced with blue, and both the C and N terminus of the BLUS1 gene are required for proper BLUS activity. Figure 1 shows that removing the BL (blue light) response reduces gs and water loss while maintaining photosynthetic carbon gain in wheat. Example 2 shows that genes homologous to Arabidopsis BLUS1 were identified in wheat: TaBLUS1-A, B and D. Wheat seeds were treated with ethyl methane sulfonate (EMS), and at least one truncation allele in each homolog resulting in a premature termination codon in the translated protein was identified. In Example 3, double and triple TaBLUS1 wheat mutants were generated using marker assisted selection. Double heterozygous A/D mutant (Aa/BB/Dd) is crossed with five different B genome mutants (AA/bb/DD), and triple heterozygous (Aa/Bb/Dd) individuals were selected and self-pollinated. These crossings produce lines with one functional TaBLUS1 copy (double mutants), no functional TaBLUS1 (triple mutant), or single mutant copy of TaBLUS1 with a predicted reduced functionality (double mutant + missense mutation). In parallel, single knockout mutants are backcrossed into elite wheat varieties (grain size or temperature tolerant phenotypes), marker-assisted backcrossing is used to generate BC3 single knockout lines which are combined into double and triple mutant combinations, and the mutant combinations are crossed into large grain size and enhanced photosynthetic rate lines. In Example 4, the impact of BL responses on double and triple mutants is assessed. Figure 1 shows that blue light-induced increases in gs reduced leaf temperature by 1-2oC. Varieties that show tolerance to high temperature and water limitation are crossed with double and triple TaBLUS1 mutants and screened for increased photosynthetic efficiency throughout the crop cycle. Photosynthetic efficiency is determined in all selected mutants/crosses to verify no adverse effects on photosynthetic capacity. In Example 5, phenotypic characterization is carried out in double and triple null mutants and the triple missense mutants in different backgrounds under yield potential field conditions.
The claimed invention lacks adequate written description for the following reasons. The instant claims are directed to a method of increasing water use efficiency (WUE) in a plant comprising reducing or abolishing the activity of a Blue Light Signaling 1 (BLUS1) polypeptide by introducing at least one mutation into a nucleic acid sequence having at least 80% sequence identity to SEQ ID NO:1, 3, 5 or 7. With regard to the wildtype BLUS1 sequences, the specification discloses the structures of BLUS1 sequences from Arabidopsis and Triticum aestivum (wheat). However, the claims encompass wildtype BLUS1 sequences obtained from sources other than Arabidopsis and T. aestivum, whereby their structures share at least 80% sequence identity to SEQ ID NO:3. The claims encompass mutants and allelic variants of the BLUS1 sequences and thus imply that structural variants exist in nature, yet no structural variant has been disclosed. The implication is that there is a gene and a protein other than that disclosed which exists in nature, but the structure thereof is not known. The disclosure of SEQ ID NO:1 isolated from A. thaliana, or SEQ ID Nos. 3, 5 and 7 isolated from hexaploid T. aestivum, is not representative of other BLUS1 sequences from other sources. Thus, there are insufficient relevant identifying characteristics to allow one skilled in the art to predictably determine such mutants and allelic variants of BLUS1 sequences from another A. thaliana, another T. aestivum, or another monocot or dicot plant, absent further guidance. Moreover, while one skilled in the art can generate a population of sequences having 80% sequence identity to SEQ ID NO:3, it is unpredictable which species within the population are wildtype BLUS1 polypeptide sequences.
The recitation of “at least one mutation” lacks adequate written description because it is unlikely that any mutation in SEQ ID NO:3 would increase water use efficiency (WUE). Applicant’s working examples teach only knockout mutants. Applicant does not address degenerate DNA and conservative amino acid substitutions. Applicant does not address mutations, the type and location, in conserved and variable regions of SEQ ID NO:3. Knockout mutations are not representative of other mutations for increasing WUE. Thus, “at least one mutation” in SEQ ID NO:3 for increasing WUE is not adequately described.
Accordingly, the claims lack adequate written description under current Written Description guidelines (Federal Register/ Vol.66, No. 4/ Friday, January 5, 2001/ Notices; p. 1099-1111. See also the Written Description Guidelines, Revision 1, March 25, 2008.)
Applicant’s Traversals
To the extent Applicant’s traversals apply to the rejection above, Applicant traverses primarily the following: (1) BLUS1 is highly conserved across the plant kingdom, and BLUS1 performs a conserved function throughout the plant kingdom due to the presence of conserved kinase and phosphorylation motifs (Fig. 1, Annex 1). (2) Claim 1 has been amended to recite introducing mutations into the nucleotide sequence encoding BLUS1 that reduce or abolish polypeptide activity. (3) It would have been routine for the skilled person in the art at the filing date to generate suitable loss-of-function mutations in BLUS1.
Response to Applicant’s Traversals
Applicant’s traversals have been considered but are deemed unpersuasive for the following reasons. With regard to traversal (1), the claims are directed to targeting wildtype BLUS1 polypeptides in a plant. However, the disclosure of the structures of BLUS1 polypeptides for Arabidopsis and T. aestivum (wheat) does not allow one skilled in the art to predict the structures of wildtype BLUS1 polypeptides from other sources. No function for the BLUS1 polypeptide is disclosed. Fig. 1 of Annex 1 has not been submitted. With regard to traversal (2), Applicant is arguing limitations not present in the claims. The scope of claim 1 indicates that any mutation would reduce or abolish the activity of a BLUS1 polypeptide. With regard to traversal (3), Applicant is arguing limitations not present in the claims. The scope of claim 1 is not limited to loss-of-function mutations in a BLUS1 polypeptide. Accordingly, the rejection is maintained.
5. Claims 1-3, 8, 11, 25 and 36 are rejected under 35 U.S.C. 112(a) or 35 U.S.C. 112 (pre-AIA ), first paragraph, as failing to comply with the enablement requirement. The claim(s) contains subject matter which was not described in the specification in such a way as to enable one skilled in the art to which it pertains, or with which it is most nearly connected, to make and/or use the invention.
Applicant’s disclosure is as set forth in the written description rejection above. The claimed invention is not enabled for the following reasons.
The scope of the claims encompasses all plants. Applicant is not enabled for all plants because the claimed method requires targeting a BLUS1 gene in a plant, and there is no evidence that BLUS1 genes have been identified in plants other than Arabidopsis and T. aestivum. The structures of BLUS1 sequences from rice, soybean or barley (claim 36) are not disclosed or known as of the filing date of the instant application. No structure and function relationship is disclosed. To require one skilled in the art to determine the structure of an unknown sequence before one can practice the claimed invention is an invitation to experiment and requires undue experimentation. Applicant does not disclose any BLUS1 encoding sequence having at least 80% sequence identity to SEQ ID NO:1, 3, 5 or 7.
The scope of “mutations” encompasses nucleotide deletions, insertions, substitutions and any combination thereof in any region of a BLUS1 sequence. Applicant discloses specific nucleotide substitutions at specific positions in the BLUS1 nucleotide sequences resulting in a STP in the encoded amino acid sequences (pp. 16-17). The specification further discloses that both the C and N termini are essential for BLUS1 activity (Example 1). However, the claims are not limited to a particular type of mutation or a mutation within a particular region of a BLUS1 encoding sequence. Neither the state of the prior art nor Applicant’s working examples shows that any mutation to a BLUS1 sequence is capable of reducing or abolishing its functional activity, especially if it is not in the C and N terminus region of a BLUS1 sequence. With regard to claim 25, there is no recitation as to what nucleotide substitution in which position of SEQ ID NO:3, 5 or 7 would result in a loss of BLUS1 activity and/or loss of stomatal response to blue light. Accordingly, Applicant has not enabled a mutation in a BLUS1 encoding polypeptide to reduce or abolish its activity and increase its WUE as commensurate in scope with the claims without undue experimentation.
In making this determination, the Office has weighed each of the Wands factors. The state of the prior art is not highly developed with regard to BLUS1 sequences, as BLUS1 sequences from only two plants are disclosed. The nature of the invention is a method to increase WUE in a plant by reducing or abolishing the activity of a BLUS1 polypeptide. The breadth of the claims encompasses all plants, known and unknown wildtype BLUS1 sequences, and all means of reducing or abolishing the activity of a BLUS1 polypeptide. The state of the prior art for increasing WUE in a plant by mutating a BLUS1 sequence is highly unpredictable, as only knockout mutations have been shown to increase WUE. The amount of direction or guidance is insufficient with regard to structural characterization of the BLUS1 sequences and mutations for achieving increased WUE. Though Applicant provides working examples, as indicated above, they are not commensurate in scope with the claims. Applicant provides insufficient direction or guidance with regard to the structures of BLUS1 sequences from different plants, ways of reducing or abolishing activity of a BLUS1 polypeptide, and mutations in a BLUS1 sequence that would reduce or abolish its activity to increase its WUE. Given these difficulties, notwithstanding a relatively high level of ordinary skill of those in the art, the amount of experimentation would likely be extensive and undue. Weighing all the Wands factors based on the totality of the record as discussed above, the Office determines that it would require undue experimentation for a person of ordinary skill in the art to make and use the invention as claimed.
Applicant’s Traversals
Applicant traverses primarily the following: (1) BLUS1 is highly conserved across the plant kingdom, the wheat and Arabidopsis BLUS1 polypeptides have conserved kinase and phosphorylation motifs, and both are part of a distinct phylogenetic clade (Fig. 1, Annex 1). (2) The genome sequences for soybean and rice were published prior to the filing date of the instant application (see Annex). (3) BLUS1 sequences from Arabidopsis and wheat include the conserved serine/threonine protein kinase domain, and this sequence information could be used to identify any suitable homologues by sequence comparison and identification of the conserved domains. (4) It would have been routine for the skilled person at the filing date to generate suitable loss-of-function mutation in BLUS1, and the mutation could be in any region of the BLUS1 sequence, so long as it reduces or abolishes BLUS1 function.
Response to Applicant’s Traversals
Applicant’s traversals have been considered but are deemed unpersuasive for the following reasons. With regard to traversal (1), Fig. 1 of Annex 1 has not been submitted. It is unclear whether BLUS1 sequences within the 80% sequence identity are in the same phylogenetic clade as Arabidopsis and wheat. With regard to traversal (2), no Annex has been submitted, and there is no evidence the BLUS1 sequences for soybean and rice are present and identified in the GenBank sequences. With regard to traversal (3), there is no evidence that the serine/threonine protein kinase domain is unique to BLUS1. Muchero et al. (US Pub. No. 20200032285 (A) teaches a Populus trichocarpa LecRLK1 (lectin receptor-like kinase 1) that has the serine/threonine protein kinase domain and is not a BLUS1 polypeptide [0089]. With regard to traversal (5), Applicant is arguing limitations not present in the claims. The scope of claim 1 indicates that any mutation would reduce or abolish the activity of a BLUS1 polypeptide and increase WUE. Accordingly, the rejection is maintained.
Conclusion
6. No claim is allowed.
7. Any inquiry concerning this communication or earlier communications from the examiner should be directed to PHUONG T BUI whose telephone number is (571)272-0793. The examiner can normally be reached on M-F 8am-5pm.
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/PHUONG T BUI/Primary Examiner, Art Unit 1663