MXPA00005751A - Constitutive plant promoters - Google Patents

Abstract

The invention describes new promoters built from elements from a set of promoters which have a complementary expression pattern.

Description

NEW VEGETABLE PROMOTERS CONS I U IVOS FIELD OF THE INVENTION The present invention relates to new plant promoters, more specifically, promoters that can be produced by assembling parts of promoters, which have a complementary specificity. PREVIOUS ART Genetic engineering of plants is now possible by virtue of two discoveries: first of all by the possibility of transforming heterologous genetic material into a plant cell (which is achieved more efficiently with the bacterium Agr ob acte ri um t urn e fa c í in so with related strains), and secondly thanks to the existence of plant promoters that are able to direct the expression of said genetic material. A typical plant promoter consists of specific elements. A base formed by the minimal promoter element, which allows the initiation of transcription, often accompanied by a sequence, also called the TATA [TATA box] sequence, which serves as REF .: 121032 binding site of the transcription start factors. In most promoters the presence of this TATA sequence is important for an appropriate initiation of transcription. Typically 35 to 25 base pairs (bp) are located towards the 5 'end with respect to the transcription initiation site. Another part of the promoter consists of elements that are capable of interacting with DNA binding proteins. The known G-sequence binding elements are those that are based on the hex anuc 1 eo t í di co CACGTG motif. It has been shown that these elements have the ability to interact with bZIP DNA binding proteins, which bind as dimers (Johnson &McKnight, Ann. Rev. Biochem., 58, 799-839, 1989). Other motifs related to G sequences have also been described, such as the Iwt and PA motifs. It has been shown that these motifs are involved in the expression of the tissue-specific promoter in plants. For example, the presence of tetramers lwt confers a specific expression in embryos, while PA tetramers confer a high level of expression in root, a low expression in leaves and no expression in seeds. Similarly, GT-1 type binding sites (grouped based on a moderate consensus sequence GGTA / TA) are described. Said binding site is located towards the 5 'end with respect to the promoter region of the promoter of the Arabidopsis and appears to be involved in the activation of transcription during periods of light (Fischer, U. et al., Plant Mol. Biol., 26, 873-885, 1994). Another phenomenon related to sequences, which is often observed in plant promoters, is the presence of sequences that allow the formation of Z-DNA. Z-DNA is DNA folded into a left-handed helix caused by repeats of GC or AC dinucleotides. It was believed that the folding in the form of Z has influence on the availability of DNA for the approach of ADU polymerase molecules, thus inhibiting the speed of transcription.

One of the first and most important inventions in the field of protein expression in plants is the use of viral (plant) promoters and derivatives of Agr ob a c t e ri um. which provides a powerful and constitutive expression of heterologous genes in transgenic plants. Several of these promoters have been used very intensively in plant genetic research and are still the promoters of choice in studies of rapid, simple and low risk expression. The most famous are: the 35S promoter and 19S mosaic virus of cauliflower (CaMV), whose practical utility was already demonstrated in 1984 (BP 0 131 623), the promoters that can be found in the T-DNA of Aqr ob a c t e ri um, such as the promoters of nopaline synthetase (nos), mannopin synthetase (mas) and octopine synthetase (oes) (EP 0 122 791, EP 0 126 546, EP 0 145 338). A promoter derived from plants with similar characteristics is the ubiquitin promoter (EP 0 342 926).

With time, an attempt has been made to increase the level of expression of these promoters. Examples thereof are the dual-potent 35S promoter (U.S. Patent No. 5,164,316) and, more recently, the superpromo t or, coupling portions of the promoters of Agr ob act eri um (EP 729 514). However, in many cases it happens that these promoters do not meet the criteria of an ideal promoter. All the promoters described above show a clear pattern of specific expression of development or organs, and often the pattern of expression obtained with these promoters is not ideal for some applications. There remains a need for new constitutive promoters capable of offering a high level of transgenic expression at the precise time and place, especially in biotechnology applications, such as the manipulation of fungal and insect resistance, which requires expression both in the location correct as in the appropriate time frame of plant development.

BRIEF DESCRTPCTM OV. TA TMVRMTTnM The invention provides new plant promoters, characteres because they comprise 1) a minimal promoter and 2) transcriptional activating elements from a set of promoters, where said elements direct a level and a complementary pattern of transcription in a plant. More specifically, this plant promoter is a constitutive promoter in which each of the activating elements of the transcript does not exhibit an absolute tissue specificity, but acts as a mediator in the activation of the transcription in most parts of the plants at a level of 21% with respect to the level reached in the part of the plant in which the transcription is most active. An example of such promoter pairs is a set of promoters in which one is more active in the green parts of the plant, while the other promoter is more active in the subterranean parts of the plant. More specifically, the new promoter is a combination of the roID promoter and the ferredoxin. It is preferred that in this construct the minimal promoter element be derived from the ferredoxin promoter and that said ferredoxin promoter be derived from Arabidopsis thaliana. The roID promoter is derived from Agrobacterium rhi z ogenes. Also forming part of the invention is a plant promoter which is a combination of the pious tocianin and S-adenosyl-methionine-1 promoter, where it is preferred that the minimal promoter element be derived from the S-adeno promoter if 1 -methionine. - 1 and that both the promoter of p 1 astocyan and the promoter S-adeno if 1 -methionine- 1 derive from A abidopsis thaliana. The invention further comprises constructs of chimeric genes for the expression of genes in plants comprising the promoters described above.

DESCRIPTION OF THE FIGURES Figure 1: Schematic representation of the plasmids pMOG410 and pMOG1059. Figure 2: Distribution of GUS expression in potato lines transformed with the constructions pMOG410 and pMOGl059. GUS staining was evaluated visually and the expression classes were measured in the laboratory in relation to the higher GUS expression (defined in 4). A value of zero indicates that there was no visible expression. Figure 3: Graphic representation of the average expression of the GUS enzyme in primary transformants of tomato, rapeseed and potatoes. The GUS expression was visually determined and compared with a GUS 35S high level transgenic tobacco plant whose score was 4. The standard deviation of the measured values are indicated in each of the bars. Figure 4: Graphic representation of the distribution of potato plants with various levels of GUS expression containing constructs SAM-1-, PC-35S and PcSAMl-GUS. The mesophilic expression of leaves, in the vascular system of leaves, stem and root was described.

DETAILED DESCRIPTION OF THE INVENTION For the purposes of this specification, the following definitions are valid. A promoter consists of a DNA polymerase binding site in DNA, which forms a transcription start site. The promoter has at least one TATA sequence and possibly other sequences surrounding the transcription initiation site (primers) and can be used either isolated (minimal promoter), or linked to binding sites of activating elements of transcription, silencers or protectors that can increase or reduce the rates of initiation of transcription and that can work with respect to the stage of development or to external or internal stimuli. The initiation site is the position that surrounds the first nucleotide that is part of the transcribed sequence, also defined as the +1 position. All other sequences of the gene and its control regions are numbered with respect to this site. The sequences towards the 3 'end (ie, other coding sequences of proteins in the 3' direction) are called positive, whereas the sequences towards the 5 'end (most of the control regions in the 5' direction) they are called negative. A minimal promoter is a promoter that only consists of all the baseline elements necessary for the initiation of transcription, such as a TATA sequence and / or an initiator. An enhancer is a DNA element that, when present in the vicinity of a promoter, has the ability to increase the rate of initiation of transcription. A promoter is constitutive when it has the ability to express the gene it controls in all or almost all plant tissues during all, or almost all stages of plant development. A specific expression is the expression of gene products limited to one or a few plant tissues (spatial limitation) and / or to one or a few stages of plant development (temporal limitation).

It is considered that there is practically no true specificity: it seems that the promoters are activated with preference in some tissues, while in other tissues they present little or no activity. This phenomenon is known as an irregular expression [leaky]. However, in this invention a specific expression means a preferred expression in one or a few plant tissues. The expression pattern of a promoter (with or without enhancer) is the pattern of expression levels that shows where in the plant and at what stage of development the initiation of transcription occurs due to said promoter. It is said that the expression patterns of a set of promoters are complementary when the expression pattern of a promoter has a small overlap with the expression pattern of the other promoter. The level of expression of a promoter can be determined by measuring the "steady state" concentration of a standard transcribed reporter mRNA. This measurement is indirect since the concentration of the mRNA informing not only depends on its speed of synthesis but also on the speed at which the mRNA degrades. Therefore, the steady state level is the product of the synthesis velocities and degradation rates. It can be considered, however, that the degradation takes place at a fixed rate when the transcribed sequences are identical, and therefore this value can serve as a measure of the synthesis rates. When the promoters are compared in this way, the techniques available to those skilled in the art are: Sl-RNase hybridization analysis, Northern blotting and competitive RT-PCR. This list of techniques does not represent in any way all the available techniques, but actually describes the procedures commonly used to analyze the transcription activity and the levels of mRNA expression.

One of the technical difficulties presented by this analysis is that qualitatively better results can only be obtained by fusing activating parts of the transcript with the informing RNA molecule, so that only the informant sequences are transcribed. This requires an accurate determination of the initiation of RNA synthesis and the binding at that point of the sequences of the informing mRNA. This is important for various reasons. First, the analysis of the starting points of the transcription in almost all the promoters has revealed that there has not been a single base on which the transcription begins, but a more or less grouped set of initiation sites, each one of the which is responsible for some starting points of the mRNA. Since this distribution varies from one promoter to another, the sequences of the informant mRNA in each of the populations differ from each other. Since each mRNA species is more or less subject to degradation, a single degradation rate for the different reporting mRNAs can not be expected. Second, in various eukaryotic promoter sequences it has been shown that the sequence surrounding the initiation site ("primer") plays an important role in determining the level of RNA expression directed by said specific promoter. This also includes a part of the transcribed sequence. Direct fusion of the promoter to reporter sequences therefore leads to very suboptimal transcription levels. If these transcribed sequences are left, the transcription rates can be determined, but the stability of the reporter mRNA is impaired and the transcription initiation rates of an eventual open reading frame are affected. The function of this analysis, however, is the determination of the relative level of the constitutive expression of a heterologous protein, the application most frequently used in biotechnology. Therefore, the most important parameter is the ability of the evaluated sequences to direct high levels of expression of a heterologous informing protein. This includes the coupling of modifying sequences of an informing protein to the activating part of the transcription, to the promoter and to the 5 'untranslated sequence of the gene evaluated for its properties. In this way a complex set of effects can be reduced (combining transcription rates, mRNA stability [and thus mRNA degradation rates] and translation initiation rates) to a value that is very useful for determine the usefulness of the gene elements evaluated in the io te cno 1 applications. There is not a word or phrase that describes this value. In the course of this application, the terms "expression level" and "transcription activity" are also used together with the term "expression value". It is considered that this may cause some confusion. In all cases, together with these and other related terms, the value just mentioned is indicated. The procedure commonly used to analyze the patterns and levels of expression is then by determining the level of "stable state" of accumulation of a protein in a cell. Candidates commonly used for the reporter gene, known to those skilled in the art, are beta-glucuronides a (GUS), chloramphenicol acetyl transferase (CAT) and proteins with fluorescent properties, as the green fluorescent protein (GFP) of A gu ora vi cto ri a. In principle, however, there are many proteins suitable for this purpose, provided that the protein does not interfere with the essential functions of the plant. For the quantification and determination of the location there is a number of suitable tools. Detection systems can be easily created or are available based for example on an immunochemical, enzymatic and fluorescent detection and quantification. Protein levels can be determined in plant tissue extracts or in intact tissues using in situ analysis of protein expression.

In general, individual transformed lines with a promoter-informing chimeric construct vary in their levels of expression of the reporter gene. It is also frequently observed that these transformants do not express any detectable product (RNA or protein), the variability in expression is commonly attributed to "position effects", although usually the molecular mechanisms underlying this inactivity are not clear. . The term average expression is used here as the average level of expression found in all lines expressing detectable amounts of reporter genes, thus neglecting the analysis of plants that do not express any detectable protein or reporter RNA. The level of expression in root indicates the level of expression found in protein extracts of whole plant roots. In the same way, "expression levels in stem" or in "leaf" are determined using complete extracts of leaves and stems. It is considered, however, that within each of the parts of the plant described above, there may be cells with variable functions, in which the promoter activity may vary. For the promoters described in this application, expression levels are measured in the large plant parts, which contain cells with diverse functions. However, a more detailed analysis can contribute to the construction of a promoter that is still "more constitutive", taking into account that more cell types are considered within a part of the plant. t Co or standard for judging expression levels, the 35S promoter of cauliflower mosaic virus constitutes a convenient and widely used standard. The average expression level of this promoter can be classified as high medium. The invention shows that it is possible to combine elements of a promoter responsible for a specific expression with elements of another promoter responsible for a complementary expression pattern, to form a promoter which, as a result, has expression in the tissues and during the stages of development that are part of the expression pattern of both promoters. If the comp emendation produces the activity in (almost) all the cells of the plant, the product of that comp i ent will be a constitutive promoter. It seems necessary, however, that both promoters have a low level of expression in the tissues and during the stages of development specific for the other promoter. It has been established that, to be adequate, the transcription activity in the parts of the plant where the expression is low is preferably > _1% of the level of transcription reached in the parts of the plant when the transcription activity is high. This limits the availability of promoters and promoter elements with which a new constitutive promoter can be constructed. Suitable pairs of promoters that meet the criteria mentioned above are: - the promoter of ferredoxin in combination with the roID promoter, the S-adenosyl methionine promoter in combination with the plastocyanin promoter. Other pairs of promoters that are complementary can also be applied, and they show at least some expression in the tissues and the specific development stages for the other promoter.

Delineation of the parts of promoters and / or potentiators necessary While regulatory elements of transcription, especially in eukaryotes, may be present at great distances from the promoter / transcription start site and may be located towards the 3 'or 5' end. 'With respect to the start site, many plant genes possess most of their regulatory elements in the area directly 5' to the promoter.

In order to identify the main activating elements of the transcription of the promoters, it is a common procedure to link parts of the non-transcribed areas found towards the 5 'end (and 3') of the promoter with the reporter gene, to analyze the ability of each of the truncated DNA elements to direct the expression of said informant. For the delineation of sequences closer to the promoter involved in the regulation of transcription, fragments of the sequences are usually coupled with the promoter, which can be derived from the gene whose transcription regulation is under study. Alternatively, a heterologous promoter, such as the 35S promoter sequences from positions 45 to +4, can be used in relation to the start of transcription, which is functionally coupled to the reporter gene as described above. In this way it is possible to delineate the activating elements of the transcription of most genes, a process well known to those skilled in the art. A large number of regulatory elements of gene transcription have been analyzed in this manner, and the data important for this analysis are available directly to those skilled in the art in scientific publications. The activating elements of transcription that on average can direct expression to approximately the average level of the 35S promoter, (at least 50% of this level) in at least some of the plant parts, and that also have the capacity of directing at least 0.5% (of the 35S level) of transcription in other parts of plants, are then selected for later use. The minimal promoter element typically derives from one of the promoters of the pair of promoters, although this is not necessarily the case. It is possible that said minimum element derives from a third promoter or also that it is produced synthetically. On the basis of the results of the analysis described above, the activating parts of the transcript are selected with complementary activities. That is, for example, a promoter that presents expression throughout the plant, transcriptional activator DNA fragments that direct a high level of expression in root and with lower levels of expression in leaf and stem, are combined with elements that direct the expression mainly in leaf and stem, but to a lesser extent in the root. Other combinations of complementary transcriptional activating parts are obvious. It is preferred that the level of expression in the parts, where the expression is lower, does not fall below 1% of the level obtained in the highest part. The situation in which the ratio between the lowest expression and the highest expression between parts of the plant is greater than 5% is more preferred. This coupling can be carried out more easily by known genetic engineering techniques. The gene to be expressed by the new constitutive promoter can be cloned behind the promoter. It is advisable to incorporate a unique Ncol cloning site at the junction of the 5 'untranslated sequence bound to the promoter to allow precise binding of the open reading frame (ORF), and the 3' end of the promoter, in which the interest can be inserted. The ferredoxin-roID pair One of the preferred combinations of the present invention is a constitutive plant promoter comprising elements of both the ferredoxin promoter and the roID promoter. It is preferred that the ferredoxin promoter is obtained from Ara b i d op s i s t h a l a where it directs the ferredoxin A gene, a gene that is involved in photosynthesis. The expression of this gene and the responsiveness of its promoter to light have been described (Vorst, 0, et al., Plant Mol. Biol., 14, 491-499, 1990; Vorst, 0. et al., The Plant J., 3 (6), 793-803. 1993; Dic'key, L.F. et al., The Plant Cell, 6, 1171-1176, 1994). Since the ferredoxin gene is involved in photosynthesis, the promoter is more active in green tissues. It was found that mRNA levels were high in organs containing cl or op 1 as t o s, such as stem, leaves and bracts, but also in young growing tissues, such as flowers and whole seedlings. Interestingly, there is a minor but significant expression in terrestrial areas of the plant. The promoter sequence has a region that contains a G sequence and a sequence I. A DNA sequence with Z-folding is also found in the -182 position. It has been reported that the roID promoter has a strong expression in the roots and can be obtained from Agroba c t eri um rhí z ogen e s. Although the organism of origin is a bacterium, the promoter is very suitable for expression in plants, because the bacterium is a phytopathogen that causes a hairy root disease in plants. For this purpose, it transfers DNA to the plant, among which the roID gene is responsible for the elongation of the root. In order to achieve its expression in plants, this gene needed a strong functional promoter in plants, the roID promoter. Studies with GUS have shown that the expression that is under the control of the roID promoter allows a root specificity to be achieved fundamentally (Leach, F. and Aoyagi, K., Plant Sci., 79, 69-76, 1991). In addition, some expression in ho j as was also observed. The combination of the ferredoxin and roID promoters can be obtained in two ways, depending on which promoter the minimal promoter element and the 5 'untranslated sequences come from. In our examples, the minimal promoter element of the ferredoxin promoter was used, but it can also be derived from the roID promoter. The pair S-adenosi 1 -methionine synthetase and pl toci aniña. Another favorable promoter can be obtained from a combination of the S-adeno s i 1 -methionine synthetase promoter (SAM) and specific parts of the promoter of the pl to to iña. It is preferred to obtain both promoters from Arabi dop s i s a l a n a. The SAM promoter regulates the expression of S-adenos i 1 -me t ionin synthetase, which is an active enzyme in the synthesis of polyamines and ethylene. Promoter studies showed strong expression in vascular tissues, in callus, sclerenchyma and some activity in the root cortex (Peleman, J. et al., The Plant Cell, 1, 81-93, 1989) probably due to the participation of the enzyme in the significance. The promoter of the parental plan, as well as the promoter of the forredoxin, is also an active promoter in the internal pathway. MRNA levels are high in green structures containing chloroplasts, such as leaves, cauline leaves, stems and whole seedlings. The promoter is also very active in flowers. Little expression can be detected in silicua, seeds and root (Vorst, 0. et al., The Plant J., 4 (6), 933-945, 1993). The combination of these specificities allows the creation of a chimeric promoter that directs a good expression in both intimal areas of leaves and stems, as well as in the areas that are not involved in said process of photosynthesis, such as in the cells that they conform and surround the vascular system in leaves and stems.

Other pairs of promoters The aforementioned examples of pairs of promoters show in both cases the presence of an active promoter during photosynthesis. It is considered that other promoters that regulate the expression of a gene necessary for the internal activity can also be suitable for a combination with either roID or other promoters with preference for the root. In the construction of a promoter that directs the expression in the whole plant: if one of the components is a promoter that is more or less specific for the green parts, this automatically means that the other promoter of the pair must be expressed predominantly (but not Exclusively) in the roots and other organs not photos in tetiz ado res. In the construction of a promoter that directs the expression in all parts of the leaves and stems, the combination can be made using a promoter that is more or less specific for the green parts, and a promoter that directs the primary expression in the vascular system.

However, the invention is not limited to the combination of a promoter with preference for the root and a promoter with preference for the green parts, and the combination of promoters with preference for green parts and the vascular system. All combinations of promoters can be used as long as the expression patterns of the individual promoters are complementary. It is also possible that the elements from which a new constitutive promoter is composed, derive from a set of more than two promoters. Therefore, there must also be the comp i emen t ar i age described above. EXPERIMENTAL PART Example 1 Cloning of the Fd-roID chimeric promoter A fragment of the ferredoxin promoter from Ara bi dop sis th aliana (0. Vorst et al., 1990, PMB, 14, 491-499) was isolated from the position - 512 to +4 (relative to the ATG start codon of the open reading frame of ferredoxin) by digestion with HindII and Ncol. This fragment contains most of the regulatory sequences of the transcription of the ferredoxin promoter, the promoter sequences and directives of the ferredoxin transcript. An Xbal site was introduced, for cloning reasons, at positions -5 to -10 relative to the ATG (0. Vorst et al., 1990, PMB, 14, 491-499). This changes the original sequence of the clone at this point from ACAAAA to TCTAGA (SEQ ID No. 1). A part of the sequences of the 5 'end roID of Agrob a c t eri um rh i z o gen e s (SEQ ID No. 2) (Leach et al., 1991, Plant Sci., 79, 69-76) to the ferredoxin promoter sequences described above. A HindlII-Rsal fragment was cloned, which comprised the nucleotides -385 to 56 in relation to the start codon, together with the ferredoxin fragment, joining the real sites of the latter with the HindIII site of the former. This chimeric element, which contained the promoter and some of the activating sequences of the ferredoxin gene, and the 5 'activating sequences of the roID gene were used in the subsequent studies due to their transcription stimulating properties (SEO ID No. 3) . Example 2 GUS Fusions The chimeric Fd-roID promoter / act i was coupled to the GUS gene, manipulated in such a way that it contained an intron gene (Jefferson et al., (1987) EMBO J 6: 3901- 3907). The Ncol restriction site at the ATG start codon was used to bind the promoter to the open reading frame (ORF) of the GUS gene, coupled to a 265 bp fragment containing transcription termination sequences, and untranslated sequences 3 'of the protease inhibitor 11 (Thornburg et al., 1987, Proc. Nati, Acad. Sci., USA, 84, 744-748, An et al., Plant Cell, 1, 115-122). The entire expression cassette, containing the GUS gene promoter, and the 3 'PI-II sequences was separated using BamHI and EcoRI and introduced into the binary vector pMOG800 (deposited in Centraal Bureau voor Schimme 1 cul tur es, Baarn, Low, with the number CBS 414.93, August 12, 1993) digested with the same enzymes. The construction obtained below (pMOG1059) was used in transformation experiments with various plants. The construction GUS-promoter CaMV 35S was used as a control. This is the construction pMOG410. In Figure 1 a schematic representation of both constructions is shown. EXAMPLE 3 Patterns and expression levels of the promoter activity, during the early stages of plant transformation First, Arabi transformants were prepared with both constructions and the GUS expression was monitored during the transformation process. The GUS expression levels were determined visually, on a scale of 0 to 5, where 0 means non-detectable expression and 5 is the highest level of GUS observed by the authors in leaves of a transgenic plant, of a rare transgenic tobacco plant 35S-GUS (line 96306). Leaf samples of this plant were included in all the experiments as internal reference. Table 1 shows the relative GUS expression in explants of Arabidopsis s thaliana r at different times after cocultivation with Agrobacterium t mefaciens (DAC-10 days after cocultivation). Table 1. Relative GUS activity of Arabi dopsi root explants Construction PMOG1059 PMOG410 DAC trial moment 0 2 3 DAC 2 3 3 DAC 5 3 3 DAC 7 4 3 DAC 9 4 3 DAC 12 4 3 As can be observed with this compilation, the GUS expression directed by the chimeric promoter starts slightly later after cocultivation, but from day 7 it exceeds the level of expression obtained with the reference 35S promoter.

Very similar data were obtained when Brassica napus explants were qualified by the GUS expression. On day 5 after cocultivation, the value of the 35S promoter was slightly higher, but the situation was reversed on day 20 after cocultivation. With tomato, similar data were obtained. Here, even at the earliest stage of the analysis, the expression of the transgenic pMOG1059 exceeded that of the transgenic pMOG410. EXAMPLE 4 Patterns and expression levels in plants grown in vitro When the plants are grown to more advanced stages, the differences between these promoters become even more evident. Samples of leaves of fully regenerated plants were analyzed by their GUS expression. Averages of 11-48 plants were obtained, according to the construction. For Arabidopsis thaliana, grown only in vitro, no great difference was observed between GUS expression in the transgenic pM0G1059 and 'pMOG410.

Table 2. Average relative GUS activity of leaf samples of all crops evaluated s Cons t ruction PM0G1059 PMOG410 Crop Potato 4.0 2.1 B a s s c a n s s a n d s 3.7 2.6 Ara bi dop s i 4.0 4.0 Tomato 2.2 2.1 From the data shown in Figure 2, it is also clear that a significant amount of GUS-35S transgenic lines (in the authors' experiments were found about 50% repetitively) does not express GUS at a visible level. Thus, only the average expression in the Fd-roID transgenics is higher, but it also strongly increases the frequency with which the transgenic plants express GUS. In approximately 50 transgenic potato plants, which contain the Fd-roID-GUS construct, the authors did not find a weak expressor, which suggests a high reliable expression in at least 98% of the elaborated lines. Example 5 Comparison of the performance of the promoter in various cultures The constructs pMOG410 (35S-GUS) and pMOG1059 (Fdro I D-GUS) were also introduced into rapeseed and tomato for further comparison of the performance of the promoter. Here we also included the data obtained with potatoes. As shown in Figure 3A, in tomatoes the general level of expression of the Fd-roID promoter is higher, both in the last stage of cultivation and in leaves of plants of 4 and 7 weeks of life. This is also observed in plant stems of 7 weeks, however in the roots a weaker average expression is observed with the Fd-roID promoter than with the 35S promoter. similar results were also obtained in rapeseed and potato, with the notable exception that in potato roots the level of expression with the Fd-roID promoter exceeds that of the 35S promoter. As shown in Figures 3B and 3C, both the average expression of the Fd-roID promoter is higher as well as the variation in expression is significantly lower. In conclusion, it can be mentioned that the authors have created a promoter that easily resists comparison with the 35S promoter in three important crops. EXAMPLE 6 Expression of the nptll transgene In order to also check the usefulness of the Fd-roID promoter for other purposes, said promoter was ligated with the nptll gene, whose expression of the corresponding gene product confers resistance in plants to the antibiotic kanamycin. This element was placed between the right and left edges of the T-DNA, and then the transformation to the plants was carried out by Agr ob a t t t um t urn e f a c i s. As a control, similar constructions were used in which the expression of the nptll gene was under the control of the nos promoter. Resistance to kanamycin in transgenic potato plants is manifested by the development of transgenic calluses and shoots, during the standard transformation procedure, in which kanamycin is used in the culture medium. In general, for constructions with the nos-nptll selection cassette, the transformation frequency for potatoes isnstructions with the selection cassette Fdro ID-np t I I the frequency is on average 61%. Although it is not known at this time how important the increase in the frequency of transformation is for this construction, it indicates that the Fd-roID promoter is at least as suitable for directing a heterologous gene, such as nptll, as the promoters. constituti usually used, as we. Example 7 Comparison of visual qualifications with quantitative values From the analysis of the GUS expression based on: 1) the histochemical analysis and the qualification with respect to an internal control and 2) the quantitative analysis of the GUS enzymatic activity, He could know that both offer reproducible quantitative results. The exhaustive analysis of both grades for leaves and roots of tomato and rape led to the conclusion that scale 3, which compares better with that of 35S, is equal to approximately 2,000 pmol MU / minut or. mg obtained in the quantitative analysis. In scale 1 and 2, the averages are 1,000 and 1,500, respectively, which defines a value of 50% for 35S. Approximately 1% expression of that level is equal to 100 pmol MU / minut or. mg, which is often below the level of detection in chemical detections, although it is sometimes detestable as a very light blue color due to GUS expression. Therefore, histochemical staining can be used as a marker to measure the effectiveness of the promoter, measuring the level of blue coloration, and using this data to select the promoter elements that will be used. Example 8 Construction of the SAM1 promoter and fusion to GUS For the construction of the SAM1 promoter (SEQ ID No. 4), genomic DNA was isolated from leaves of Arabi dop s i s t h a l a to Landsberg erect using the CTAB extraction procedure. Primers were designed on the basis of the published sequence of the SAM1 gene from Arabi dop s i s t h a l a a K05 (Peleman et al., (1989) Gene, 84, 359-369). The promoter element was amplified by PCR (30 cycles of 45 seconds at 95 ° C, 45 seconds at 50 ° C and 1 minute at 72 ° C, this program was used in all the other PCRs described in this part) using the primers FR-Ps am- 143: 5 'AGA TTT GTA TTG CAG CGA TTT CAT TTT AG 3' (SEQ ID No. 5) and FR-Psam-216- 5'ATC TGG TCA CAG AGC TTG TC 3 '(SEQ ID No. 6) and an approximately 550 bp fragment was obtained. The DNA fragment was isolated from an agarose gel and cloned into the pGEM-T vector (Promega Corp., Madisan Wl, USA). This clone was used as a template to introduce an Ncol site at the start of the translation by PCR using primers FR-Psam-144: 5 'GTC TCC ATG GTG CTA CAA AGA ATA G 3' (SEQ ID No. 7) and FR-Psam-143. The resulting 500 bp fragment was cloned into the vector pGEM-T. The EcoRI and HindIII sites located in the promoter region were removed by PCR in two Steps using this clone as annealed. In this PCR, the BamHI site was introduced 5 'with respect to the SAM1 promoter and the HindIII site was introduced at the 3' site of the promoter. In the first step of the PCR, three promoter fragments were generated. The first fragment (1) contains the 5 'BamHI site and the EcoRI site mutated using the primers FR-Psam-248: CGG GAT CCT GCA GCG ATT TCA TTT TAG_3_' (SEQ ID No. 8) and FR-Psam-249 : 5 'AC TGA ACG AAT GCA AAA TCT C 3' (SEO ID No. 9). The fragment of the medium (2) is obtained with the primers FR-Psam-250: 5 'AGA TTT TGC ATT CGT TCA TGT G 3' (SEQ ID No. 10) and FR-Psam-251: 5 'TGT AAG CAT TTC TTA GAT TCT C 3 '(SEO ID No. 11). This fragment has a partial overlap with fragments 1 and 3 and has mutated sites EcoRI and HindIII. The third PCR fragment (3) contains the mutated internal HindIII site and a HindilII site is introduced at the 3 'end of the promoter that encompasses the Ncol site at the start of translation and is generated using the FR-Psam primers. 252: 5 'AAG AAA TGC TTA CAG GAT ATG G 3' (SEO ID No. 12) and FR-Psam-253: 5 'GAC AAG CTT GAT CCC ATG GTG CTA CAA AGA ATA G 3' (SEO ID No. 13 ). In a second PCR the three fragments were mixed 1, 2 and 3 in a tube and amplified with the primers FR-Psam-248 and FR-Psam-253. Due to the superposition between fragments 1 and 2 on the one hand and 2 and 3 on the other, this PCR allows obtaining the complete mutated promoter. After digestion with Ba HI and HindIII, the resulting SAM1 promoter was cloned into a pBSK + vector. The SAM1 promoter was then cloned into a vector containing the GUSint rón-TPT-I I reporter cassette exchanging the 5 'region using the BamHI and Ncol restriction sites. This was effected by digestion of the SAM1 clone with Ba Hl and Ncol and isolation of the promoter fragment from an agarose gel. The GUS vector was digested with the same enzymes, and the vector was then isolated from an agarose gel, whereby the promoter of the original 5 'sequences was discarded. The SAMl-intron promoter cassette GUS-TPTII was then cut from the vector by digestion with BamHI and EcoRI after which the reporter cassette was isolated from an agarose gel and cloned into the binary vector pMOG800 digested with BamHI and EcoRI. . The resulting binary vector PMOG1402 was introduced into strain EHA105 of Agrob a c t e ri um t um efa ci in s for transformation into potato. Example 9 Construction of the chimeric promoter Ps-SAMl and fusion with the GUS gene The enhancer of p 1 as t o ci aniña (Pe) of Arabi dop s i a th i a n a Col-o was obtained by PCR. To this end, the primers FR-Pc-146 (5 'agt ggt acc atc ata ata ctc atc ctc ett a 3') were developed (SEO ID No. 14) and FR-Pc-247 (5 'cga age ttt here aat cta att tea tea cta aat cgg a 3 ') (SEO ID No. 15) introducing a 5' Kpnl restriction site with respect to the enhancer and a HindIII restriction site with respect to the plastocyanin buffer. PCR was carried out using pfu DNA polymerase (Stratagene) for 30 cycles of 1 minute at 95 ° C, 1 minute at 50 ° C, 4 minutes at 72 ° C and a cycle of 1 minute at 95 ° C, 1 minute at 50 ° C, 10 minutes at 72 ° C. The resulting fragment of the PCR was ligated into a large copy number cloning vector using Kpnl and HindIII, resulting in the construction of pPM15.1. This clone was used as a template for PCR (30 cycles of 1 minute at 95 ° C, 1 minute at 50 ° C, 2 minutes at 72 ° C) using the primers FR-Pc-145: 5 'GCT GCA ATA CAA ATC TAA TTT CAT CAC TAA ATC GG 3 '(SEQ ID No. 16) and FR-Pc-145: 5' AGT GGT ACC ATC ATA ATA CTC ATC CTC CTT C 3 '(SEO ID No. 14). The PCR generates an approximately 850 bp fragment comprising the Pe enhancer (SEQ ID No. 17) which contains a 5 'Kpnl site and overlaps the 5' end of the SAM1 promoter (see Example 6). The PCR fragment was then mixed with the PCR fragment of the SAM1 promoter generated with the primers FR-Psam-143 and FR-Psam-144 using the pBKS + clone containing the adjusted SAM1 promoter described in Example 8. In a PCR With this mixture, the PcSAM chimeric promoter was generated using the primers FR-Psam-144 and FR-Pc-146. The resulting promoter fragment of approximately 1.3 kb (SEQ ID No. 19) was isolated from an agarose gel after digestion with KpnI and Ncol and then cloned into a large copy number cloning vector (pUC28) digested with the same enzymes. The promoter fragment was cut out of this vector by digestion with BamHI and Ncol and cloned in front of the GUS intron gene described above in Example 8. The complete Pc-SAM-GUS-TPT-11 reporter cassette was then cloned into the pMOG900 as described for the SAM1-GUS-TPT-II reporter cassette in Example 8. The resulting binary vector pMOG1400 was introduced into the EHA105 strain of Agrobacterium tumefaciens for its transformation into potato.

EXAMPLE 10 Construction of the Pc-35S enhancer promoter and fusion with the GUS gene. The enhancer of pineapple protein (Pe) from A abidopsis thaliana Col-0 was obtained by PCR (see above). This clone was used as a template for a PCR (30 cycles of 1 minute at 95 ° C, 1 minute at 50 ° C, 2 minutes at 72 ° C, the other PCR reactions described in this part were carried out with the same program) using the primers FR-Pc-291: 5 'GTC TTG TAC AAA TCT AAT TTC ATC ACT AAA TCG G 3' (SEQ ID No. 19) and FR-Pc-146: 5 'AGT GGT ACC ATC ATA ATA CTC ATC CTC C 3 '(SEQ ID No. 14). PCR generates an approximately 850 bp fragment comprising the enhanced Pe containing a 5 'Kpnl site and overlapping the 5' end of the 35S minimum promoter. The minimum 35S promoter was obtained in a PCR using pMOG971 as an annealed containing the 35S promoter and 5 'omega OTR) and the primers FR-35s-292: 5' TTA GAT TTG TAC AAG ACC CTT CCT CTA TAT AAG G 3 ' (SEO 10 NO: 20) and lsl9 (SEO ID No. 19). The resulting fragment is superimposed with the Pe enhancer and contains an internal Ncol site at the start of the translation. Then the two PCR fragments were mixed and the PCR reaction was carried out using the primers FR-Pc-146 and lsl9. The resulting fragment was then digested with Kpnl and Ncol, isolated from an agarose gel and cloned into pUC28 digested with the same enzymes. The resulting clone was then digested with BamHI and Ncol and the promoter fragment (SEO ID No.22) was isolated from an agarose gel and cloned 5 'with respect to the GUS gene as described in Example 7. A Then the complete informant cassette was introduced into the binary vector pMOG800 as described in Example 7. The resulting binary vector pMOG1401 was introduced into strain EHA105 of Agr ob acte ri um t um e fa ci s for transformation into potato.

E xemplo 11 Patterns and expression levels in plants grown in vi tro Transformed plants were cultivated and leaf samples of plants completely regenerated by their GUS expression were analyzed. Figure 4 shows the analysis of leaf mesophyll expression, leaf vascular system, stems and roots. A very low level of GUS staining was observed in the mesophilic part. of the leaves of SAMl transgenic plants, although the rating indicates a GUS expression level of 0.

LIST OF SEQUENCES (1) GENERAL INFORMATION: (0 APPLICANT: (A) NAME: ZENECA MOGEN (B) ADDRESS: Einsteinweg 97 (C) CITY: Lei in (E) COUNTRY: The Netherlands (F) ZIP CODE: 2333 CB (G) TELEPHONE: (31) 71-5258282 (H) TELEFAX: (31) 71-5221471 (ii) TITLE OF THE INVENTION: New constituent plant promoters (iii) QUANTITY OF SEQUENCES: 22 (iv) COMPUTER READING FORMAT: (A) TYPE OF MEDIA: DISKET (B) COMPUTER: IBM COMPATIBLE PC (C) OPERATING SYSTEM: PC-DOS / MS-DOS (D) SOFTWARE : Patentin Reeléase N ° 1.0, Version Na 1.25 (EPO) (vi) DATA OF THE PREVIOUS APPLICATION: (A) NUMBER OF APPLICATION EP 97203912J (B) DATE OF DEPOSIT: DEC 12, 1997 (2) INFORMATION OF THE SEQ ID ": 1: (i) CHARACTERISTICS OF THE SEQUENCE; (A) LENGTH: 520 base pairs (B) TYPE: nucleic acid (C) STRING: double (D) TOPOLOGY: linear (ii) TYPE OF MOLECULE: DNA (genomic) ) (iii) HYPOTHETIC: NO (ii) ANTI-SENSE: NO (i) DESCRIPTION OF SEQUENCE: SEQ ID NO: 1: oACTGstITTO rc ?? í? JTc? fc GfcrrKrtW? T SC? JTTTC-TTS ?? ttaBTAift < a, ttc? *, tot «o SWG &TÍCíft? * TfttTTRaBTT ATOTJlATt: ftJ-TOWOT? TTTACRStR TTO0M3TTA? A. 120 ATrararrTfJ? R TCSOTO-aerE •? S? »CC | (-) IT? ß-auu '? A? Cccßßeíra CMS? C? TO? Eo TAC? FiCt?? L? R CC? CJW KT -? TWVrrr? I GTSQftftCAftA CSPUtftfCT »TI? ÍM-ACrj? T a-tfl acnAM_AftÃO TCttSW-W-M; TOTCAÚC? GT COdXWT? FtG J? CtóMWW 'ACTC? CCTCA 390 JWNM-AT? CW OTSCTOWAS TCiracciuw; aG ?? rc * QM: MWG ^ CBAW C? TAAMl C i So A * »?«? KAGA TCCKTCTA? CCM ??????????????????????????????????????????? cftAseewfft? tecrcftAAAA 4ÍO? GCTCCCM? AC? A? TCT «» »* cc * fc 5 * 3 (2) INFORMATION OF SEQ ID N °: 2: (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 300 base pairs (B) TYPE: acid nucleic (C) CHAIN: double (D) TOPOLOGY: linear (ii) TYPE OF MOLECULE: DNA (genomic) (iü) HYPOTHETIC: NO (iii) ANTI-SENSE: NO (xi) DESCRIPTION OF SEQUENCE: SEQ ID N °. 2: xxx ?? ríiC ?? K-? AtrTd? T stsf? Sox • oesttaaßet? cerflßacAtc c? ¿05- * eíre so TGC < wc «tt? rj? TATsccc Tßoocec? cc tßAficcc3 ntTCGCWOSfl? t »t e * p» \ a or ccccscRTT? A C-Vrrajwtntt ßxcsSfiMNW ßcciOCAC *. cBKdftrtßB a? awcRCWi HD d € * t < cetem lAcmCia * ccßeaar? ooc ACJC WV? SW ww AAcßCQ «COCXCROC 24 * 1 OCaCT TCCA aACOtftGßC ß-MTCWM? T GCSCXßaCAC JiAM? MT »» KS * ft ?? trr 300 (2) INFORMATION OF SEQ ID Nß: 3: (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 840 base pairs (B) TYPE: nucleic acid (C) CHAIN: double (D) TOPOLOGY: linear (ii) ) TYPE OF MOLECULE: DNA (gepomic) (iii) HYPOTHETIC: NO (iii) ANTI-SENSE: NO (xi) DESCRIPTION OF SEQUENCE: SEQ ID NO: 3: OG? TCCQ? ßC «OCATOtiCC CX? Cl? Cftftr tS? ÜS tQtnx. «TO? CJ TT JdStt? COOtX < ? crraeexTee 9 «PNICCXCS ßoßßcigd¡i-r t» tjurtccer «3GceÉ &cc? TBA? CCO? T? uo trccc x 3fT TIOTCMTAC JCCOWTÍ-? C ATtewo? tM tcftn»! roa9 ccroaujaic? ßr OWGOiiíftCG C? GßCTCfiAC TCMKS? Wt ACGGGC? I? C CGG? TCaaC C * tt «trsTA 2 * 0 OAWtxcocaa ACCCKCHÉCB «crncc **« COOT-MOÍ ct «aw» KR > ca iaaeíak soo? MU-A_RXA. tcc «Awsts ÍPTCI? OTOT GJW»? G «AÍ? AT? VT3 »TG cac m i'cn. 3 co TTI-UCT? GAC at? -? wctft * - * FTCA? TT rrmsrai TATJ-MR? T? ATG < - st? "or TCTiCMST? AT TßCGT? AAA TO®rrT0? Tt CßCT-? TtiTT GRTAO Í? T OC? MRTr? IC CCrt-OWMu »A« VT «? TOrt ncíACtaffTC CACATCrrtT * TGfitptACf BSsuiCftJU r S4C ßXAC? srr? T iWKmeßMX CM? cwmt CÍ? -? T CT citßAScrare COUCTM-M 6th « CCACGTAIVTA CTCXC TOA CAAO? MÍ? G T CtfX? AET CTCTCÍ? MCJv CA? TCACXA CAC? CVS? TC ATM ?? CAC *? AS? OVkTA? TCCWCC »TC C? CASM G? -23CC * rtSTC crrAsieencaii txcre? Icn» RAWG? CIC * acvrtruixs tttc? Oa? Cß cavmcc? 'ßo.

AfcOCCMJWT OC CJAft AT CTCT? ÍTTCA ICrCaeaJi? A GSCJ? RXCr »SaO? SoWGC # 40 (2) INFORMATION OF SEQ ID N °: 4: (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 477 base pairs (B) TYPE: nucleic acid (C) CHAIN: double (D) TOPOLOGY: linear ( >) TYPE OF MOLECULE: DNA (genomic) (iii) HYPOTHETIC: NO (ffl) ANTI-SENSE: NO (V) ORIGINAL SOURCE: (A) ORGANISM: Arabidopsis thaliana (Xi) DESCRIPTION OF SEQUENCE: SEQ ID NO. : «CATCcroc *. aoaxrrrcxr txXMS ttcr awuut »at tawwp * rß? 8ß? rr an 00 < Day »ßtrtn wpm aw ??? aatxB waimaGC« aRrrmo Aawrrroa. 120 t «: GTtc? -ro T wwatam iW icHTtGTíwc rrasjAtGftT t? M? Tctr? CIT? AATOT 130 GC? -C? C? O? pumuu- wr? CQUWO ?? «AcaMeca. «Master? Ciw. ermynio UA? AAB? TA C? C? WkMR • MTrOMWTO XtTX3 »ßC2T« WSJKSC * * ß * 0CtrWr wrtct5CTs JUU-TXCGWT c noi iTOS TpMCtí? e AürcTWGß. A TXJAQ ?? TC TA? GAAAWK.- TTACAdCuLíA TOCWJ-AACT AftCOTOTU KX? SCKTGh TOCT t-TTTT AiaitctC? Ac Aoiacc? AXß? «Trttttt mÂrtcrat TCnrcn.cc? CCA? SC? 7 (2) INFORMATION OF SEQ ID Np: 5: (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 29 base pairs (B) TYPE: nucleic acid (C) CHAIN: simple (D) TOPOLOGY: linear (ii) ) TYPE OF MOLECULE: cDNA (iíi) HYPOTHETIC: NO (xi) OESCRIPTION OF THE SEQUENCE: SEQ ID NO: 5: AWWfTTOT *? T * cCT * Tr MtTTM? B 2 * (2) INFORMATION OF SEQ ID N °: 6: (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 20 base pairs (B) TYPE: nucleic acid (C) CHAIN: simple (D) TOPOLOGY: linear ( I) TYPE OF MOLECULE: cDNA (ni) HYPOTHETIC: NO (xD DESCRIPTION OF SEQUENCE: SEQ ID NO: 6: ATCTßßTotc? - * - 3Ctrt «c 20 (2) INFORMATION OF SEQ ID N °: 7: (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 25 base pairs (B) TYPE: nucleic acid (C) CHAIN: simple (D) TOPOLOGY: linear ( ii) TYPE OF MOLECULE: cDNA (iii) HYPOTHETIC: NO (xi) DESCRIPTION OF THE SEQUENCE: SEQ ID NO. 7: ctcTcavsßG TßcTACswAß *? »To 35 (2) INFORMATION OF SEQ ID ND: 8: (0 CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 27 base pairs (B) TYPE: nucleic acid (C) CHAIN: simple (D) TOPOLOGY: linear (ü) TYPE OF MOLECULE: cDNA (Mi) HYPOTHETIC; NO (xi) DESCRIPTION OF THE SEQUENCE: SEQ ID NO: 8: caj-saxcep? C * nex »trre 27 ATT? SC (2) INFORMATION OF SEQ ID NO: 9: (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 22 base pairs (B) TYPE: nucleic acid (C) CHAIN: simple (D) TOPOLOGY: linear ( i¡) TYPE OF MOLECULE: cDNA (iii) HYPOTHETIC: NO (xi) DESCRIPTION OF THE SEQUENCE: SEQ ID N °: 9: 2? (2) INFORMATION OF SEQ ID N °: 10: 0) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 22 base pairs (B) TYPE: nucleic acid (C) CHAIN: simple (D) TOPOLOGY: line! (I) TYPE OF MOLECULE: cDNA (üi) HYPOTHETIC: NO (xi) DESCRIPTION OF SEQUENCE: SEQ ID NO: 10: Aca m-rCC * reotfriCh3ti is 3-1 (2) INFORMATION OF SEQ ID NO: 11: (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 22 base pairs (B) TYPE: nucleic acid (C) CHAIN: simple (D) TOPOLOGY: linear ( ii) TYPE OF MOLECULE: cDNA (iii) HYPOTHETIC: NO (xi) DESCRIPTION OF THE SEQUENCE: SEQ ID Na: 11: Tc swascatóT TCGT? ISWTC te 28 (2) INFORMATION OF SEQ ID N ° 12: (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 22 base pairs (B) TYPE: nuefeic acid (C) CHAIN: simple (D) TOPOLOGY: linear (ii) ) TYPE OF MOLECULE: cDNA (iii) HYPOTHETIC: NO (xi) DESCRIPTION OF SEQUENCE: SEQ ID N °: 12: AAC &AKtset "MCWK3MAT 62 za (2) INFORMATION OF SEQ ID NO: 13: (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 34 base pairs (B) TYPE: nucleic acid (C) CHAIN: simple (D) TOPOLOGY: linear ( ii) TYPE OF MOLECULE: cDNA (iii) HYPOTHETIC: NO (xi) DESCRIPTION OF THE SEQUENCE: SEQ ID NO: 13: cyvC? WícrTG. Hrccc? XOGr OCTAOWAOJI. AT? C 34 (2) INFORMATION OF SEQ ID NO: 14: (0 CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 31 base pairs (B) TYPE: nucleic acid (C) CHAIN: simple (D) TOPOLOGY: linear (¡) TYPE OF MOLECULE: cDNA (iü) HYPOTHETIC: NO (xi) DESCRIPTION OF THE SEQUENCE: SEQ ID NO: 14:? GKXMt? Cfc TC? TAATACT 31 (2) INFORMATION OF SEQ ID NO: 15: (9 CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 37 base pairs (8) TYPE: nucleic acid (C) CHAIN: simple (D) TOPOLOGY: linear (« ) TYPE OF MOLECULE: cDNA (iü) HYPOTHETIC: NO (xO DESCRIPTION OF THE SEQUENCE: SEQ ID NO: 15. ctyyurmprTA CARWCWAIcncncn MTCTOH 31 (2) INFORMATION OF SEQ ID NO: 16: (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 35 base pairs (B) TYPE: nucleic acid (C) CHAIN: simple (D) TOPOLOGY: linear TYPE OF MOLECULE: cDNA (ñl) HYPOTHETIC: NO (xi) DESCRIPTION OF THE SEQUENCE: SEQ ID N »: 16: OCIOC? AI? C AAAICTAATG TC? TC? CT? TGCC 3S (2) INFORMATION OF SEQ ID NO: 17: (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 876 base pairs (B) TYPE: nucleic acid (C) CHAIN: double (D) TOPOLOGY: linear ( ií) TYPE OF MOLECULE: DNA (gepomic) (iii) HYPOTHETIC: NO (iv) ANTI-SENSE: NO (vi) ORIGINAL SOURCE: (A) ORGANISM: Arabidopsis thaliana (xi) DESCRIPTION OF SEQUENCE: SEQ ID NO: 17: XlOhOtaKC tCa-? OCWC «SrC? Ü * T? WHCWEiVrc? YOlUMCT AOTA« WACT «OR * to rt? nt aiiuaBvtr? t * tta. * avrr oaatc *** AAMXSWT? A -j? ea «wnr 120 AAtfSWTBKO «ATATÉWIIA ACTTA? AtpT? $ 08 £ ftGM iTT? ATTIOS XATaATAATT IßD 9TCTACT AÍI CTTJSlíWftlC TXKXC? XMK AUCCTTAOTO rnwtt3SCAA TACri rcAA aio sT? Nw? TOia QAKGCATAI.G arccchuro • «? WJÍ ?? T TT-ATCOMA-C viATrccxavr 3 QD 7T QTWn SfcTW? CCW. IWCTtWK? CA < ICCATTA SCTATA? D? -Í TTOGTTCACA Jíú »« 3 A GGT «? TA? -? A?»? WWAICWG sirpetTAT «M * M * e * t« swAari? - 70 ? .TACA »3dCA CACXrrKJSrC OftBrtTCM? A ßCCAJ ?? ITTA CM, TMIAArj (J k? NOEAT DO -MTCGTATC Tapw? AAGt T-? Ssirassc TCNWOKXFC TGAAOJOCCÍ TC? TATCAT $ * e -tpascocíioe ccoßccovp; Aicair? Ct-a? Oajcrßttc TA ?? C ?? TGC enroll »ßoo TATOGTOAM. arßccMCSws MXCOGSGOOT paaovAwi? GDOG GAAO ß «c QAAGft? OcQA Atr »ca-M At uteoep jMS-xst Ax. ? tt * rrcrr TOCTßMßAU. 7S0 AT OßMßC TOUUWCfccr TOIC AAC J-ftßSBOftenS? JrrX? Ofi? A * tXU? 06 * 780 GAATC6 O6A sOWWC? CC TACp = »* 6? T? OEAWW? TA * CADATtATCT TXTU? H? SST «4C TACOTA? WB. ? -rc? fGUUvr nßATWßi O-BUWT «75 (2) INFORMATION OF SEQ ID NO: 18: (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 1357 base pairs (B) TYPE: nucleic acid (C) CHAIN: double (D) TOPOLOGY: linear ( ii) TYPE OF MOLECULE: DNA (genomic) (iii) HYPOTHETIC: NO (iv) ANTI-SENSE: NO (xi) DESCRIPTION OF SEQUENCE: SEQ ID NO: 18: as vreecci? G OI * O VCC ** juwjstíttaírc CTCCTTCTCA A-WGTCCTAC CTATTMO? *or TATC? WSS? T MJttWt? TCr TC? T? TW? T? Crm? XTC * TCMWTt8Q ACAAAAAAT6 130 A.tr wsetct? TßíTKfctew, ptaetxxm we-WMír? * TWK ?? üwß. c * f? B? T5A * sa? TGGCCTA SII. «KGR? K? TA qrmeatrpt wa crotc XTKU? ACCS wraottiw to» «OCt? TACTT TTCWWna? S? »OM-n; ATUCtBl C CWírßtserT ATSmaCTG 3 ©? wA? AtctAit tßSATtrr < ? r TOG8IWJ? B? eCGKTQfcc? wß? CG »ß? C cw»? * ßctxt aso ? ym * .nt.txn: «cwcxjPßa-A jam * r i? nsMSwtcffe icríwßtwß CGX? TOWT? 420 tn ecwia AT «NMRC *. ? SOC? CWKW "tsMccsißrr? T? CSßGcAT JWFT bed # iO cAAoaca-Aw TOMwweG l? -w aßstt MASTUAOT lßooracM» oWtettfAC S * Ó Go -oitertT TtaaBWßsu CCWCOOTOC naaonan: eacawuxQc twrrctA? Au «00 AATSCO0FVO T? CCCT? TOT TW-WSU? ftaSAOftCßßC ßiantTATO TAA? A0ATOK Í4U Ar M? CA? AC OC? Fl- WUSGA tCCSA &? N? C C3COWWCCCA TG? CAATTTG AOCSOft? Os 7t) wcmwc'm A3 * »* GMOCTC? JA ocniAtc CCAACAWWI < &«j« -trr? BC XCQMKCTTA iraos? OAieKr exeavkenitf CTCVCXKO JACB? CCAJW oß c? AT a * c T? R rAA .juxnrtwcca ATrt? ST? Ftr wqyfft * a * .t KSTKT? TiC ßCGVtrrCRT '900 tt? * IS? Rtßr CAMC IA G cte * a *? 6TG tßßß? Trt < ? < wi-o? tfrrttA tctwccTK? sit dCArcwiw M¡wßts? «» c «Ac« rtt? w á »a« t? tacA tresnaos to * otAt 5 one TCA-ÍWOIftpß rtßATATßAT TWU «C1« r-- WSTIkA rt »K-VBiGACJA T? FtßTAAC? T Í0.BD ? CQAWTAS C TCACtAAe? TAITÍAOTCM. CTjWtIT? ßC TWUU? C? TA TCACTTA? A? 3.1 * 1) "TTcaUKrc ATrrr? rrr Tm? vraucr? BC? TJWG QAVICTKJU TAATTISB? TG tzoo ü wriürs vtsM &tcrc arrerpM * arpwawrtc? w? SM« tac TWttaßotoA 1 * 50"SOxiWkACt ATFCtm? A ßAtMO-AUNT •" XWL- '.IT nSGKTttpcJt AGIOBC? AO 1? 20 mrtCmr rctu racc wtrm fr * APW ug? (2) INFORMATION OF SEQ ID N °: 19: (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 34 base pairs (B) TYPE: nucleic acid (C) CHAIN: simple (D) TOPOLOGY: linear. { ? \) MOLECULE TYPE: cDNA (iii) HYPOTHETICAL: NO (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 19:? ERCP z Cft AAXCTARTW CA-Pocta A "" 3 * (2) INFORMATION FOR SEQ ID NO: 20: (0 SEQUENCE CHARACTERISTICS: (A) LENGTH: 34 base pairs (B) TYPE: Addò nudeico (C) STRANDEDNESS: single (D) TOPOLOGY: linear ( ") MOLECULE TYPE: cDNA ( iii) HYPOTHETIC: NO (i) DESCRIPTION OF THE SEQUENCE, SEQ ID NO: 20: tE * »twirr * a? μj? CcCt tr? fcra, tw ** ac Í« (2) INFORMATION OF SEQ ID N °: 21: 10 (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 19 base pairs (B) TYPE: nudeic acid (C) CHAIN: simple (D) TOPOLOGY: linear (H) TYPE OF MOLECULE: cDNA (iü) HYPOTHETIC: NO (Xi) DESCRIPTION OF THE SEQUENCE: SEQ ID NO: 21: 3 a2c * s? CA cskcmrsc 13 (2) INFORMATION FOR SEQ ID NO: 22: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 1006 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: double (D) TOPOLOGY: linear 00 TYPE OF MOLECULE: DNA (genomic) HYPOTHETIC: NO (iv) ANTISENT? DO: NO (xi) DESCRIPTION OF THE SEQUENCE: SEQ ID NO: 22: «S ?? CGCCCC« ACOÍTCAT -XW? OATC tSKSTTTCTC? -WßTWTAC SXATTWl? * ßo TAiciMa? T «mcrtßtcf« ffP! Rc «s? T? ctrmm? I go-look? AAMAAIC 120 ATTAAGOICT tSfflTIUWBS-A TtXWTKt * T "TaAVtCrßi tATTWaßG» OCl-JIXJM xa or mw-T TG »íATK? CBi otheriT? Ta« rcwwwrc? WOIAACCT »e ??? rt? A.t 40 CGCAATA GT TGCAWW? W. ísmjp pwapñ AJBMTOG? CC Cacra "* rr r junanme 300 CagiAÍCTA" SWTTTKff 1xM "W" ** ACCOATßA G teaACtt 'C C ßlWßlJAT 360 SAG? TTIO0T TCAGF.TMUUI? TTTTrtAT? JK? TAATGI? T tíMOMTTQ CttMG ?? T? "twenty • tA ATOTßAT ATGGA? TACA TBOCAC? QOT tCerrt? * ßtt TC? CWK- WÍ AWJGACX? W < *OR SAAdsCAAAT TCC? TTßtGC TfMCTdß ?? UMKTJUSir TGOSCTC? ßA C? TTCtWAC? 0 ßGccncrtT ATCATTCOOC CCASCCCKWC cc Tcserc? T CKTAACOCC TGITCTAAAG eor- AATocaprß nccn¡? Xßt Tßuuwrreßs noßsc cccc GToa peAse TSU? PG? CQA teo AOft? GCAßAC ßC? OAGAAOA? CSMrTTAC CCCAAftCCQA? C8QI? QO? R. -rTAfaATTT 750 TTC-TTOCTO AtßUWGAT »FUßCTCM *? TiAtt AfC CCMlCAAOm. GftßfcO? CTTr 783 ACOAAACITA XCOeACAMC OTOCAGCAAT CTCWT * tK? ACftAGC? AT flßThA A? AT «« TAifCrritW? CCgfFaCQB JTOrnCTßJlT CAJATTASAT rtStfO? O? CCCTtCCm üoo AXRxrAoaAA CGGCATITCA tttßw »aA £ H AC? Oßnmr TGJWAACAAT TACCSACAAC« cs AACAAAC? AC MACMCATT? OUn-r? CT? TTTM3U.TT? OCATGC lßoB It is noted that in relation to this date, the best method known to the applicant, to carry out the aforementioned invention is that which is clear from the manufacture of the objects to which it relates.

Having described the invention as above, the content of the following is claimed as property:

Claims (15)

1. CLAIMS 1. A plant promoter, characterized in that it comprises a minimal promoter and transcription activating elements from a set of promoters, where the elements present a complementary pattern and level of transcription in the plant.
2. 2. The plant promoter according to the rei indication 1, characterized in that each of the activating elements of the transcription, do not exhibit a specific tissue specificity, but act as mediators in the activation of the transcription in most parts of the plant to a level > _ 1% of the level reached in the part of the plant with the highest transcription activity.
3. 3. The plant promoter according to claim 1 or 2, characterized in that one of the promoters, of the set of promoters, is specifically active in the green parts of the plant, while the other promoter is specifically active in the subterranean parts of the plant
4. 4- The constitutive plant promoter according to rei indication 3, characterized in that it is a combination of the ferredoxin promoter and the RoID promoter.
5. 5. The constitutive plant promoter according to claim 4, characterized in that the minimal promoter element is derived from the ferredoxin promoter.
6. 6. The constitutive plant promoter according to claim 4 or 5, characterized in that the ferredoxin promoter is derived from Arabi dop si s th a l a n a.
7. 1 . The constitutive plant promoter according to claim 6, characterized in that it comprises the sequences of the SEO ID No. 1 and SEQ ID No. 2.
8. 8. The constitutive plant promoter according to claim 7, characterized in that it comprises the sequence of the SEO ID No. 3.
9. 9. The constitutive plant promoter according to claim 3, characterized in that it is a combination of the p-tocyanin promoter and the S-adenosyl-t-onin-1 promoter.
10. 10. The constitutive plant promoter according to claim 9, characterized in that the minimal promoter element is derived from the promoter of S-adeno s i 1 -me t ionin-1.
11. 11. The constitutive plant promoter according to claim 9 or 10, characterized in that the promoter of the plant is derived from Arabidopsis s thaliana.
12. 12. The constitutive plant promoter according to claim 9, 10 or 11, characterized in that the promoter of S-adenos il-met ionin-1 is derived from Arabidopsis s thali ana.
13. 13. The constitutive plant promoter according to claim 12, characterized in that it comprises the sequences of the SEO ID No. 4 and the SEO ID No. 17
14. 14. The constitutive plant promoter according to claim 13, characterized in that it comprises the sequence of SEQ ID No. 21.
15. 15. A chimeric gene construct for the expression of genes in plants, characterized comprises the promoter of any of the rei indications 1-14.