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 .
Status of Claims
Claims 1-12 and 14-28 are pending.
Claims 5, 12, 14-26 remain withdrawn for being directed to a non-elected invention(s).
Claims 8-11 remain withdrawn for being directed to a non-elected species.
Claims 1-4, 6-7, and 27-28 are examined in this Office Action.
The text of those sections of Title 35, U.S. Code, not included in this action, can be found in a prior Office Action.
Response to Amendment
The Declaration under 37 CFR § 1.132 filed on 12/11/2025 is acknowledged. The Declaration does provide evidence that the unique properties of N. tabacum make it an unusual grafting partner for tomato.
Information Disclosure Statement
Initialed and dated copy of Applicant’s information disclosure statement (IDS) filed on
12/11/2025 is attached to the instant Office Action. The submission is in compliance with the provisions of 37 CFR 1.97. Accordingly, the information disclosure statement is being considered by the examiner.
Claim Rejections - 35 USC § 103
Claims 1-3 and 27-28 remain rejected under 35 U.S.C. 103 as being unpatentable over Albacete (Albacete et al., 2009, Plant, Cell & Environment, Vol. 32, pp. 928-938; included in non-Final rejection dated 12/11/2024) in view of Ruiz (Ruiz et al., 2020, Frontiers in Plant Science, Vol. 11, pp. 1-19), Seda (Seda et al., 2020, Acta Agriculturae Slovenica, Vol. 115(2), pp. 297-305), and Yasinok (Yasinok et al., 2009, Journal of the Science of Food and Agriculture, Vol. 89(7), pp. 1122-1128).
Claim 1 recites “a method for producing a non-naturally occurring composite tomato plant comprising an abiotic stress tolerance, the method comprising:
providing as a rootstock an allotetraploid tomato plant comprising the abiotic stress tolerance;
providing as a scion a Solanum lycopersicum variety; and
grafting the scion on the rootstock, thereby producing the non-naturally occurring composite tomato plant comprising the abiotic stress tolerance.”
Albacete teaches recombinant inbred lines (RILs) derived from a cross between S. lycopersicum L. var. cerasiforme × S. cheesmaniae (L. Riley) Fosberg were used as rootstocks (i.e., providing as a rootstock a tomato plant comprising an abiotic stress tolerance), and the commercial tomato hybrid cv. Boludo F1 used as the scion (Sc). Grafting was performed using the splicing method (i.e., producing a non-naturally occurring composite tomato plant comprising an abiotic stress tolerance) (Albacete, page 929, Materials and Methods, first paragraph).
It is known in the art that the hybrid ‘Boludo’ is a commercial tomato Solanum lycopersicum (i.e., providing as a scion a Solanum lycopersicum variety).1
Albacete teaches that wild tomato species have often been considered a useful source of salt tolerance genes (i.e., an abiotic stress tolerance) to transfer to the cultivated tomato (Albacete, page 928, Introduction, first paragraph). As is the case of Solanum peruvianum , Solanum cheesmaniae , Lycopersicon pimpinellifolium, Lycopersicon hirsutum, and Lycopersicon pennellii. Moreover, this tolerance can also be transferred to a more sensitive cultivar by using these genotypes as rootstocks (i.e., a non-naturally occurring composite tomato plant comprising an abiotic stress tolerance) (Albacete, page 928, Introduction, second paragraph).
Albacete does not explicitly teach providing as a rootstock an allotetraploid tomato plant comprising an abiotic stress tolerance.
Ruiz teaches that climate change is already challenging agriculture. It will result in higher temperatures, and increased soil salinity. As a major force for plant evolution, polyploidy promotes better adaptation traits in crops, since polyploid plants are thought to have been selected during evolution because of their phenotypic and genomic plasticity (Introduction, page 2, right column, first full paragraph).
Ruiz teaches that polyploidy is one of the main factors driving evolution in higher plants, conferring genotypic plasticity by increasing the number of copies of the genome (autopolyploidy) or adding different genomes (allopolyploidy), thus increasing their potential for adaptation and promoting their selection. It has been proposed that polyploidy favors adaptive evolution to changing environmental conditions through differential expression of duplicate genes (Introduction, page 2, left column, first paragraph).
Ruiz teaches that in agriculture, the genomic modifications that take place during polyploidization confer many interesting advantages over the diploid (2x), for example, enhanced tolerance or resistance to abiotic stresses (Introduction, page 2, left column, third paragraph).
Ruiz teaches that polyploidy, whether auto or allo, is associated with enhanced tolerance to a wide range of stresses, including salinity and heat, both in wild and cultivated plant species. In artificial allopolyploids, research has shown that it is possible to efficiently combine the desired parental phenotypes on the progenies, although genome instability and allelic losses have also been described. Overall, polyploid breeding is progressively carving out its place as a method to improve crops for abiotic stress tolerance, as it opens the possibility of adding functional novelty, while combining genomes that are associated with a well-known and highly valued agronomic behavior. The outcome minimizes the risk that undesired behavior causes economic loss when compared to traditional breeding methods (page 10, left column, second full paragraph).
Ruiz teaches that in vegetable crops, grafting has been used mainly since the beginning of the 20th century. Grafting of a scion or crop variety onto a rootstock is effective at providing abiotic stress tolerance and resistance. Thus, combining grafting impacts with the use of a polyploid rootstock and scion might bring great advantages in cultivated crop (Introduction, page 2, left column, fourth paragraph). In grafted or composite crops, benefits can be provided both by the rootstock’s adaptation to the soil conditions and by the scion’s excellent yield and quality. Thus, grafted crops provide an extraordinary opportunity to exploit artificial polyploidy, as the effects can be independently applied and explored at the root and/or scion level, increasing the chances of finding successful combinations. The use of synthetic tetraploid (4x) rootstocks may enhance adaptation to abiotic stresses in perennial crops (i.e., providing as a rootstock an allotetraploid tomato plant comprising an abiotic stress tolerance) (Abstract, page 1).
It is known in the art that Nicotiana tabacum (tobacco, 2n = 4x = 48) is a natural allotetraploid combining two ancestral genomes closely related to modern Nicotiana sylvestris and Nicotiana tomentosiformis.2
Similarly, it is also well known in the art that tomato as a scion has been successfully grafted onto tobacco rootstock. See, for example, the teachings below.
Seda teaches that the rising world population increases the demand for agricultural products. In order to supply this growing demand, research on faster and more inexpensive plant production methods to enhance quality, yield potential, and tolerance against stressful conditions have been enhanced. Grafting enables the cultivation of agricultural products in different climates and soil conditions by utilizing benefits from the characteristics of different rootstocks. Besides vegetative reproduction, the yield of plants resistant to biological and environmental stress without damaging product quality positively effects crops, and has become a method of producing plants with broader ecological tolerance (Introduction, page 298, first paragraph).
Seda further teaches the effects of grafting tomato on different tobacco rootstocks on quality factors and nicotine content. The commercial variety (Solanum lycopersicum ‘H2274’) of tomato was used as the scion plant, and six different tobacco (Nicotiana tabacum L.) rootstocks were used: Taşova, Tekel, Muş, Samsun, Dişbudak, Hasankeyf cultivars. Yield of non-grafted and grafted plants grown in open-field conditions was calculated, and there was a significant increase in yield in grafted tomatoes compared to nongrafted tomatoes (Abstract, page 297).
Furthermore, Yasinok teaches that grafting is considered an established technique for crop production. Grafting of vegetables onto resistant rootstocks facilitates resistance of scion against soil-borne disease, high and low temperatures, and high salt concentration. Moreover, one of the main objectives of grafting is to increase yield. Influences of rootstock on scion resistance against harsh conditions and diseases as well as on productivity and quality are crucial in determining the potential use of grafting applications (Introduction, first paragraph).
Yasinok further teaches two different tomato scions, cv. Elazig and cv. Sweet (cherry) (Solanum lycopersicum L.) were grafted onto tobacco root stock (Nicotiana tobacum L.). Tobacco grafting had a positive effect on the tomato plant cultivation performance; the onset of flowering was almost 15 days earlier and the tomato flower and fruit yields increased in both tomato cultivars. Tobacco grafting resulted in 5.0% and 30.1% increase in total fruit weight for cv. Sweet and cv. Elazig, respectively (Abstract).
At the time the instant application was filed, it would have been obvious and within the scope of
one of ordinary skill in the art to utilize the tomato grafting method as taught by Albacete using an allotetraploid rootstock as taught by Ruiz, Seda, and Yasinok to arrive at the method for producing a non-naturally occurring composite tomato plant comprising an abiotic stress tolerance as required by instant claim 1. Although Albacete, Ruiz, Seda, and Yasinok do not explicitly teach an allotetraploid tomato plant comprising an abiotic stress tolerance, one would have been motivated to produce an allotetraploid tomato plant comprising an abiotic stress tolerance to use as the rootstock in the grafting method as taught by Albacete knowing that allopolyploidy is associated with enhanced tolerance to a wide range of abiotic stresses, including salinity and heat, as taught by Ruiz, Seda, and Yasinok. Indeed, Albacete, Ruiz, Seda, and Yasinok all teach grafting in fruit-bearing vegetables as a common procedure to develop abiotic stress tolerant plants in which both rootstock and scion affect abiotic stress tolerance. Thus, one of ordinary skill in the art would have a high expectation of success by combining the teachings of Albacete, Ruiz, Seda, and Yasinok.
The method of producing non-naturally occurring composite plants comprising an abiotic stress tolerance by grafting a scion on an allotetraploid rootstock comprising an abiotic stress tolerance is a technique that was routine in the art at the time the application was filed, as taught by the cited references and the state of the art in general.
In regard to claim 2, Albacete teaches rootstock derived from a cross between S. lycopersicum L. var. cerasiforme × S. cheesmaniae (L. Riley) Fosberg (i.e., species from group Esculentum) (Albacete, page 929, Materials and Methods, first paragraph).
In regard to claim 3, Albacete teaches the commercial tomato hybrid cv. Boludo used as the scion (i.e., the scion is a commercial variety) (Albacete, page 929, Materials and Methods, first paragraph).
In regard to claims 27 and 28, Ruiz teaches that allopolyploidy is associated with enhanced tolerance to a wide range of stresses, including salinity and heat, both in wild and cultivated plant species. (i.e., wherein the non-naturally occurring composite plant is salinity tolerant; wherein the non-naturally occurring composite plant is heat tolerant) (page 10, left column, second full paragraph).
Claims 4 and 6-7 are rejected under 35 U.S.C. 103 as being unpatentable over Albacete (Albacete et al., 2009, Plant, Cell & Environment, Vol. 32, pp. 928-938) in view of Ruiz (Ruiz et al., 2020, Frontiers in Plant Science, Vol. 11, pp. 1-19), Seda (Seda et al., 2020, Acta Agriculturae Slovenica, Vol. 115(2), pp. 297-305), and Yasinok (Yasinok et al., 2009, Journal of the Science of Food and Agriculture, Vol. 89(7), pp. 1122-1128) as applied to claims 1-3 and 27-28 above, and further in view of Ghani (Ghani et al., 2020, The Journal of Horticultural Science and Biotechnology, Vol. 95(4), pp.506-520; included in Non-Final rejection dated 12/11/2024) and Chauvin (Chauvin et al., 2003, Plant Cell, Organ, and Tissue Culture, Vol. 73, pp. 65-73; included in Non-Final rejection dated 12/11/2024).
Claim 4 further requires “applying a chromosome doubling agent to the interspecific hybrid plant, or a vegetative cutting thereof, to generate a chimeric interspecific hybrid; growing the chimeric interspecific hybrid to produce a tomato fruit; collecting seed from the tomato fruit; and growing the seed to produce an allotetraploid tomato plant comprising an abiotic stress tolerance”.
The combination of Albacete, Ruiz, Seda, and Yasinok does not explicitly teach applying a chromosome doubling agent to the interspecific hybrid plant to generate a chimeric interspecific hybrid.
Ghani teaches that interspecific hybridization is an important method in the plant breeding program which improves the different crop species. It can incorporate useful genes into cultivated genotypes which are responsible for desirable characteristics or traits in many important field crops. Solanaceous crops like tomato, brinjal, peppers, potato, and tobacco have great range of wild species which have excellent source of desired characters that can be used in crop improvement (Introduction, page 506, left column, first paragraph).
Ghani further teaches that tomato (Solanum lycopersicum L.) is facing a lot of challenges against production, such as abiotic stress, which cause the reduction of yield. Abiotic stress such as temperature, frost, heat, and drought severely damage its quality and yield. The use of genetically resistant cultivars could be an economical and environmentally friendly approach to control against abiotic stress. These problems can also be solved by transferring resistance genes from wild species to S. lycopersicum species through breeding method such as interspecific hybridization. Wild relatives of cultivated tomatoes including S. pennelli, S. peruvianum and S. pimpinellifolium have resistance genes against salinity stress (Introduction, page 506, left column, second paragraph).
Ghani teaches plant genotypes comprising Solanum pimpinellifolium, Solanum lycopersicum, Solanum Arcanum and Solanum pennellii. Seeds of genotypes were sown; crossing was made between all genotypes in all possible combinations (i.e., crossing a Solanum lycopersicum variety with a plant from an abiotic stress-tolerant Solanaceae species to produce interspecific hybrid seed). Absent evidence to the contrary, the Solanum lycopersicum and the Solanum pimpinellifolium used in the crosses taught by Ghani are essentially homozygous (i.e., the Solanum lycopersicum variety and the plant from the stress-tolerant Solanaceae species are essentially homozygous). After fruit setting and ripening, fruits were collected; fully ripened tomatoes were crushed and seeds were collected (i.e., produce interspecific hybrid seed); hybrid F1 plants were grown (i.e., growing the interspecific hybrid seed to produce an interspecific hybrid plant) (Ghani, page 507, Materials and Methods, first paragraph).
Ghani does not explicitly teach applying a chromosome doubling agent to the interspecific hybrid plant to generate a chimeric interspecific hybrid.
Chauvin teaches that a potato breeding scheme implies the possibility of ploidy level manipulation either by reducing the chromosome number of cultivars from 48 to 24 to be able to cross them with diploid related species or by doubling diploid material to reach the generally optimal tetraploid level. Since oryzalin has proven to be efficient as a chromosome doubling agent on potato cell suspension cultures, this herbicide was tried on various Solanum species and interspecific diploid hybrids. Fifty–100% of the regenerated tetraploid plants acclimatized after in vitro treatment proved to be chimaeric (Chauvin, Abstract, page 65).
Chauvin teaches that utilization of wild species is useful since it allows introgression of many desirable traits such as disease and pest resistances or quality traits. Many methods have been explored to obtain 4x interspecific hybrids, such as chromosome doubling of the 2x parent prior to carrying out the cross at the 4x level using colchicine treatment of the apical buds in situ (Chauvin, Introduction, page 65, first paragraph).
Chauvin teaches crossing several S. tuberosum dihaploid x diploid related species (i.e., interspecific hybrid plant); growing the material to obtain fully developed plantlets; submitting apical shoots of the plantlets to various Oryzalin treatments (i.e., applying a chromosome doubling agent to the interspecific hybrid plant, or a vegetative cutting thereof, to generate a chimeric interspecific hybrid); rooting the explants and growing for a complete vegetative cycle (i.e., generate a chimeric interspecific hybrid); tubers were then harvested (i.e., growing the chimeric interspecific hybrid to produce a fruit) and planted after conservation (i.e., collecting seed from the fruit) , constituting families of daughter-plants issued of the same initial doubling event (i.e., growing the seed to produce a stress-tolerant allotetraploid plant) (Chauvin, Materials and Methods, pages 66-67).
In regard to claims 6 and 7, Ghani teaches plant genotypes comprising Solanum pimpinellifolium, Solanum lycopersicum, Solanum Arcanum and Solanum pennellii. Seeds of genotypes were sown; crossing was made between all genotypes in all possible combinations (i.e., species from group Esculentum; selected from S. galapagense, S. cheesmaniae, S. lycopersicum, S. pimpinellifolium, and hybrid combinations thereof) (Ghani, page 507, Materials and Methods, first paragraph).
At the time the instant application was filed, it would have been obvious and within the scope of
one of ordinary skill in the art to utilize the chromosome doubling method as taught by Chauvin using the plant genotypes Solanum pimpinellifolium and Solanum lycopersicum as taught by Ghani to arrive at the method for producing a stress-tolerant allotetraploid tomato plant as required by instant claim 4. Although neither Chauvin nor Ghani explicitly teach a stress-tolerant allotetraploid tomato plant, one would have been motivated to produce a stress-tolerant allotetraploid tomato plant to use as the rootstock in the grafting method as taught by Albacete knowing that allopolyploidy is associated with enhanced tolerance to a wide range of abiotic stresses, including salinity and heat, as taught by Ruiz. Thus, one of ordinary skill in the art would have a high expectation of success by combining the teachings of Albacete, Ruiz, Ghani, and Chauvin.
The method of producing an allotetraploid tomato plant by using a chromosome doubling agent is a technique that was routine in the art at the time the application was filed, as taught by the cited references and the state of the art in general.
Response to Applicant’s Arguments
Applicant's arguments filed 12/11/2025 have been carefully considered but they are not persuasive.
Applicant argues that the mere existence of the claimed elements in prior art is not sufficient to render an invention obvious. A reason for combining or modifying must also exist, and there is no reason to combine the cited art.
The Examiner respectfully disagrees. Although Albacete already teaches rootstocks having an abiotic stress tolerance, Ruiz further teaches that grafted crops provide an extraordinary opportunity to exploit artificial polyploidy, and the use of synthetic tetraploid (4x) rootstocks may enhance adaptation to biotic and abiotic stresses in perennial crops such as apple or citrus (Ruiz, Abstract, page 1). Ruiz goes on to teach that polyploidy is one of the main factors driving evolution in higher plants, conferring genotypic plasticity by increasing the number of copies of the genome (autopolyploidy) or adding different genomes (allopolyploidy), thus increasing their potential for adaptation and promoting their selection. It has been proposed that polyploidy favors adaptive evolution to changing environmental conditions through differential expression of duplicate genes. In agriculture, the genomic modifications that take place during polyploidization confer many interesting advantages over the diploid (2x). Ruiz further teaches that combining grafting impacts with the use of a polyploid rootstock and scion might bring great advantages in cultivated crops. Climate change will result in higher temperatures, drought, and increased soil salinity. As a major force for plant evolution, polyploidy promotes better adaptation traits in crops (Ruiz, Introduction, page 2). Thus, Ruiz teaches an enhancement of adaptation to biotic and abiotic stresses which will be necessitated by climate change.
Applicant further argues that there is no reason to modify the cited art.
The Examiner respectfully disagrees. Applicant emphasizes that “it is noted three times in this single paragraph of Ruiz that additional research into polyploidy is required”. It is noted that Ruiz is a review article that comprehensively covers both the successes and the failures of using polyploid rootstocks and scions in grafting. A fuller reading of Ruiz would include the following excerpt:
Overall, polyploid breeding is progressively carving out its place as a method to improve crops for abiotic stress tolerance, as it opens the possibility of adding functional novelty, while combining genomes that are associated with a well-known and highly valued agronomic behavior. The outcome minimizes the risk that undesired behavior causes economic loss when compared to traditional breeding methods. In the following section, we review the effect of genome duplication on abiotic stress tolerance. Specifically, we will focus on the biochemical, morphological, and physiological modifications underpinning the enhanced tolerance of polyploid crops to a wide range of environmental stresses that have been described lately (page 10, right column, first paragraph).
Ruiz cites the following examples to support this statement: autotetraploid “Rangpur” lime has higher constitutive production of ABA than the 2x counterpart, associated with increased drought tolerance; stomatal closure in 4x Arabidopsis is more responsive to drought and ABA than in 2x; “Carrizo” citrange maintains unaltered leaf hydric status under osmotic stress, allowing gas exchange parameters to be sustained and limiting water consumption, while the 2x is drastically affected; 4x Acacia (Acacia senegal L. Willd) grew faster than 2x only under drought stress; and, finally, polyploid Arabidopsis has greater tolerance to salinity, associated with higher K+ uptake and lower Na+ accumulation in leaves. Surprisingly, this effect has been shown to rely on rootstock polyploidy rather than on shoot cytotype. Hence, it is a root-dependent phenotype that could be provided to grafted crops using polyploid rootstocks. Higher K+ retention when faced with salt stress has been also observed in hexaploid bread wheat (T. aestivum L.), in “Carrizo” citrange, and in the allohexaploid sweet potato wild relative Ipomoea trifida (Kunth) G. Don (Ruiz, pages 11-12).
Therefore, these two things may be true at the same time – additional research into polyploidy is required on more crops, and there have been numerous documented successes of using polyploid rootstocks in grafting to enhance biotic and abiotic stress resistance.
Applicant argues that the success of various grafting combinations with Nicotiana does not convey a reasonable expectation of success for tomato.
The Examiner does not dispute the assertion that Nicotiana is an atypical grafting partner, or that pepper/tomato and eggplant/tomato are generally not compatible. The obviousness rejection is based on the suggestion that allotetraploid tomato rootstock can be successfully grafted onto tomato scion, improving abiotic stress resistance. By combining the teachings of Albacete, Ruiz, Seda, and Yasinok, one of ordinary skill in the art would learn that tomato crop productivity under salinity can be improved by grafting cultivars onto salt-tolerant wild relatives (Albacete); polyploidy, whether auto or allo is associated with enhanced tolerance to a wide range of stresses, including drought, salinity, cold, heat, nutrient deprivation, or excess light both in wild and cultivated plant species (Ruiz); grafting tomato/tobacco practices have significantly positive effects on tomato yield and quality (Seda); and, tomato–tobacco grafting is a novel and promising technique for improvement of not only tomato plant performance and yield, but also that it can be employed to various tomato varieties (Yasinok). Taken together, one of ordinary skill in the art would have a high expectation of success by following the teachings of Albacete, Ruiz, Seda, and Yasinok.
Applicant argues that the allotetraploid rootstocks of the present disclosure are superior over the closest interspecific hybrid diploid rootstocks.
The Examiner respectfully disagrees. In light of the teachings in Albacete and Ruiz, one of ordinary skill in the art would expect the allotetraploid tomato rootstock to be superior over the closest interspecific hybrid diploid rootstocks. Albacete demonstrated the successful grafting of salinity resistant wild tomato rootstock onto tomato scion, and Ruiz teaches the enhancement of desired traits by using an allotetraploid rootstock. Thus, one would expect an improvement in salinity resistance by using an allotetraploid rootstock.
In regard to Applicant’s remarks regarding compact prosecution, the Examiner respectfully disagrees with the characterization of “piecemeal examination” and “moving the proverbial carrot”. The Non-Final Rejection dated 12/11/2024 was based on the claim set dated 10/17/2024, which recited a “stress-tolerant composite tomato plant”. The subsequent claim set dated 03/11/2025 amended claim 1 to require “a composite plant comprising an abiotic stress tolerance”, and added new claims 27 and 28 requiring “salinity and heat tolerance”. These amendments required a new field of search for abiotic stress (salinity and heat) tolerance, leading to the new prior art (Ruiz) in the Final Rejection dated 04/07/2025. A Request for Continued Examination (RCE) was filed on 07/07/2025, which essentially re-opens prosecution and resets the examination process allowing the Examiner to reiterate or strengthen the positions in the previous rejections. The Examiner believes that compact prosecution has been exercised and there has been no “piecemeal examination” or “moving the proverbial carrot” in this case.
Summary
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.
Correspondence
Any inquiry concerning this communication or earlier communications from the examiner should be directed to CHRISTINA MEADOWS whose telephone number is (703)756-1430. The examiner can normally be reached Monday - Friday 9:00 am - 5:00 pm.
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CHRISTINA MEADOWS
Examiner
Art Unit 1663
/CHRISTINA L MEADOWS/Examiner, Art Unit 1663
/Amjad Abraham/SPE, Art Unit 1663
1 See evidence by Flores (Flores et al., 2010, Scientia Horticulturae, Vol. 125, pp. 211-217; included in non-Final rejection dated 12/11/2024).
2 See evidence by Skalická (Skalická et al., 2005, New Phytologist, Vol. 166(1), pp. 291-303).