JP3200627B2 - Plant promoters specific for meiotic cells - Google Patents

Description

DETAILED DESCRIPTION OF THE INVENTION

[0001]

TECHNICAL FIELD The present invention relates to a plant promoter having a specific promoter function in meiotic cells and a method for using the same.

[0002]

2. Description of the Related Art A promoter is a signal existing on a genomic DNA sequence and defining the start of transcription (mRNA synthesis). A start point called a TATA box required for the start of transcription is present near and upstream of the transcription start point, and it is considered that this site is necessary for the transcription start point. In gene recombination technology, a strong promoter is useful for artificially introducing and expressing a target gene. For example, cauliflower mosaic virus (CaM
The 35S promoter of V) has the property of being constitutively expressed and has been frequently used so far.

[0003] However, when extremely limited temporal and spatial regulation of gene expression is desired, it is necessary to use a tissue-specific and time-specific promoter. Conventionally,
A pollen-specific promoter has been reported as a promoter sequence that controls specific gene expression in germ cells, but no promoter showing specific expression in meiosis has been found. It has been desired to obtain and use a specifically expressed promoter.

Up to now, only one example of a promoter of a gene expressed in meiosis, the AtDMC1 gene of Arabidopsis has been reported (Klimy).
ukand Jones, Plant J. 11: 1-
14 (1997)), its expression specificity is not strict, showing very high expression levels in roots, and has also been shown to be expressed in mitotic somatic cells (Doutriaux).
et al. , Mol Gen Genet 25
7: 283-91 (1998)). On the other hand, there have been studies in which the promoter of mei2, a meiosis-specific gene of fission yeast, is transiently expressed in plant cells and its function is observed, but they do not use transformed plants or the like. This is a finding in cells, and no finding at the tissue or organ level has been obtained at all (Tabata et a
l. , Plant Sci 8931-41 (199
3)).

[0005]

An object of the present invention is to provide a promoter that is specifically expressed in meiotic cells of higher plants. That is, the present invention is to isolate a promoter that specifically activates transcription only in meiotic cells during meiosis, which has not yet been discovered or developed yet, and to construct a foreign gene system using the promoter. If such a meiosis-specific promoter is developed, it can be a basic technology related to artificial control of plant reproduction or genetic recombination. It is a further object of the present invention to provide a method for isolating a promoter sequence from a large genome organism and a method for tissue-specific and time-specific expression of a foreign gene in a transformed plant using the same.

[0006]

[MEANS FOR SOLVING THE PROBLEMS] LIM (Lily Me
sages Induced at Meiosi
s) Genes are genes expressed in pollen mother cells, and are known to exhibit timing specificity and tissue specificity (Kob)
ayashi et al. (1994) DNARe
s. 1, 15-26). Isolation and characterization of a group of genes having such specificity is possible only in the lily of the valley where uniform meiotic progression can be observed. Therefore, an expression control region (promoter) located upstream of the LIM gene group
It is thought that the acquisition of will lead to elucidation of the characteristics of the promoter responsible for its specificity. In addition, since phenomena such as meiosis or pollen formation are considered to be conserved in higher plants, promoters isolated from lily-of-thorn lilies may show similar timing and tissue specificity in plants belonging to other taxa. Be expected. Based on the above findings, a promoter located upstream of the LIM gene group was collected from the pollen mother cell of Lily of the valley. As a result, evaluation of transcriptional activation by measuring luciferase activity showed an effect of specifically causing transcriptional activation in meiotic anthers.

[0007] The promoter of the present invention is the first meiosis-specific promoter in plants. As described above, promoters of various plants have been isolated and their expression specificities have been examined. However, no promoter has been reported that specifically expresses only in meiotic cells. The DNA fragment of the present invention has extremely strict temporal and spatial expression specificity as a meiosis-specific promoter, and is considered to be useful also in that respect. Specific uses of the present invention include the production of male sterile plants, etc., but it is also expected to be the beginning of the development of basic technologies that allow artificial control of plant reproduction or genetic recombination. .

[0008]

BEST MODE FOR CARRYING OUT THE INVENTION The plant promoter specific to meiotic cells of the present invention is specified by the nucleotide sequences shown in SEQ ID NO: 1 in the sequence listing and SEQ ID NO: 2 in the sequence listing. According to the genetic recombination technique, a specific site of a basic DNA can be artificially mutated without changing or improving the basic characteristics of the DNA. A gene having a natural nucleotide sequence provided by the present invention, or a gene having a nucleotide sequence different from the natural one, similarly artificially inserted, deleted, substituted by the same as the natural gene Alternatively, it may have improved properties, and the present invention includes such a mutant gene. In addition, the plant promoter specific to meiotic cells revealed in the present invention is the longest sequence obtained by the following experimental method. Range. That is, as long as the plant promoter of the present invention specifically has a plant promoter function in reduced cells, a part of the nucleotide sequence described in SEQ ID NO: 1 in the sequence listing and SEQ ID NO: 2 in the sequence listing is partially deleted or replaced. Alternatively, a substituted plant promoter is also included.

Also, a method for transforming a plant by introducing the plant promoter of the present invention into a plant, an expression vector having the plant promoter introduced therein, a transformed plant obtained by introducing the plant promoter into a plant, The plant seed derived from the transformed plant obtained in the above is also within the scope of the present invention. By introducing the plant promoter of the present invention into a plant, it is possible to specifically activate transcription in meiotic germ cells. Examples of plants into which the plant promoter of the present invention is introduced include monocots such as lily, rice, corn, asparagus, and wheat, and dicots such as Arabidopsis, tobacco, carrot, soybean, tomato, and potato. . As a method for producing a transformant, a usual method known in the art can be used. Vectors that can be used in the present invention are plasmid vectors, and include, for example, pBI121 and pBI221 used in Examples, but are not limited thereto. By introducing such a vector into, for example, Agrobacterium and infecting a callus or a young plant, a transformed plant can be produced, and further, a seed derived from such a transformed plant can be produced. It is possible to get The transformation method for introducing the plant promoter of the present invention into a plant is not limited to the Agrobacterium method, and a method such as a particle gun method and an electroporation method can also be used.

[0010]

EXAMPLES (Extraction of DNA) DNA was extracted from pollen mother cells of Lily of the valley by the phenol / chloroform method. That is, according to the method of the report of Murai et al. By the CTAB method (Murray, GC and Thomp).
son, W.S. F. (1980) Nucl. Acid
Res. 8,4321-4325), and DNA was extracted. Specifically, sea sand and liquid nitrogen were added to about 1 to 1.5 ml of pollen mother cells, and the mixture was ground well with a mortar and pestle.
ml of 2% CTAB was added and further homogenized, which was incubated at 65 ° C. for 30 minutes. Next, chloroform / isoamyl alcohol (24: 1)
Was added and mixed by inversion for 5 minutes. It was centrifuged at 8000 rpm for 20 minutes in a Beckman JA-20 rotor to obtain a supernatant. The resulting mixture was dispensed into a microcentrifuge tube in a volume of about 800 μl, and 1 ml of 1% CTAB was further added and mixed by inversion. After standing at room temperature for 1 hour, 1 minute at 8000 rpm
After centrifugation for 0 minutes, a precipitate was obtained. Then, 600 μl of 1 M CsCl was added, and the mixture was heated to 45 ° C. to dissolve the precipitate. Then add 1.2ml of 100% ethanol and add 12000rpm
centrifugation for 5 minutes to obtain a precipitate. It was rinsed with 70% ethanol, dissolved in 600 μl of TE buffer, further treated with Rnase, and subjected to phenol / chloroform treatment three times. After ethanol precipitation, 300 μl of TE
To obtain DNA.

(Primer Design) In order to obtain a promoter, information on the nucleotide sequence of the cDNA of the LIM10 and LIM18 genes showing meiosis-specific expression (Kob
ayashi et al. (1994) DNA Re
s. 1,15-26) were synthesized. The base sequence of the primer used is shown below. 10Pro1: ACGCTGGCGACTTCGAGTCGAACCGTCAG (2
9mer) 10Pro2: GCGGCGAGCAAAGTATCATATTCCTCTCTT (3
0mer) 18Pro1: ATCATTGGCTTGTCACGGGTGCCGGAGATC (3
0mer) 18Pro2: GGTCGCTGAAGTGACGGCCGATCAGCCGC (2
9Pro) 18Pro3: ATGACTCGCGCACTCTCTGGATCGTTCGAG (3
0mer) 18Pro4: AGTGAGGTCGCCATGGTTCTCCGTTGCGAG (3
0mer)

(Acquisition of LIM10 Promoter) A genomic DNA serving as a template is treated with BamHI and a cassette attached to the kit is added. Then, a polymerase chain reaction (P) is performed using cassette primer C1 and primer 10Pro1.
CR). PCR was heated at 94 ° C for 5 minutes,
The reaction was carried out under the conditions of 30 cycles of 94 ° C. for 30 seconds-64 ° C. for 1 minute-72 ° C. for 2 minutes, and stored at 4 ° C. after completion of the reaction. Further, using the cassette primer C2 and the primer 10Pro2, PCR was performed under the conditions that after heating at 94 ° C for 5 minutes, a reaction was performed at 94 ° C for 30 seconds-66 ° C for 1 minute-72 ° C for 2 minutes for 30 cycles, A DNA fragment was obtained. This DNA fragment was obtained using pT7Blue T (manufactured by Novagen).
And cloned into the vector by the TA method. As a result of analyzing the base sequence, the base sequence described in FIG. 1 and SEQ ID NO: 1 in the sequence listing, which is the upstream sequence of LIM10, was obtained.

(Acquisition of LIM18 Promoter) A genomic DNA serving as a template is treated with EcoRI to add a cassette attached to the kit.
PCR was performed using 8Pro1. After heating at 94 ° C for 5 minutes, the reaction was carried out under the conditions of 30 cycles of 94 ° C for 30 seconds-62 ° C for 1 minute-72 ° C for 2 minutes for 30 cycles. Furthermore, cassette primer C2
And 18Pro2 at 94 ° C for 5 minutes, then 94 ° C
The PCR was carried out under the conditions of performing 30 cycles of the reaction at -62 ° C for 1 minute and the reaction at 72 ° C for 2 minutes for 30 seconds to obtain a DNA fragment. This DNA fragment was obtained using pT7BlueT-
The vector was cloned into the vector by the TA method, and the nucleotide sequence was determined.

Further, based on the obtained base sequence information,
After two kinds of primers 18Pro3 and 18Pro4 were synthesized and the genomic DNA was treated with XhoI and the attached cassette was added to a kit (LA PCR ^{™ in} vitro cloning kit manufactured by Takara Shuzo), cassette primers C1 and C1 were added.
After heating at 94 ° C for 5 minutes using 8Pro3,
PCR was performed under the conditions of 30 cycles of a reaction at 0-66 ° C. for 1 minute-72 ° C. for 2 minutes, and stored at 4 ° C. after completion of the reaction. In addition, cassette primers C2 and 18Pro4
After heating at 94 ° C for 5 minutes, the mixture was heated at 94 ° C for 30 seconds-66.
PCR was carried out under the condition of 30 cycles of a reaction at 1 ° C. for 1 minute and a reaction at 72 ° C. for 2 minutes to obtain DNA fragments.

The DNA fragment was blunt-ended using a DNA branding kit manufactured by Takara Shuzo, and the 5 ′ end was phosphorylated using polynucleotide kinase manufactured by Takara Shuzo.
It was digested with SmaI and cloned into a dephosphorylated pBluescript SKII + vector manufactured by Stratagene. As a result of determining the nucleotide sequence, the nucleotide sequence shown in FIG. 2 and SEQ ID NO: 2 in the sequence listing, which is the upstream sequence of the LIM18 coding region, was obtained.

(Construction of vector) By inserting the DNA fragment of the present invention shown in SEQ ID NO: 1 and SEQ ID NO: 2 in the sequence listing upstream of DNA having an arbitrary base sequence, its meiosis cell-specific transcription is performed. The product can be expressed. In this embodiment, tobacco (Nicotiana)
tabacum cv. The firefly luciferase gene was introduced into SR1). Plant vectors pBI121-LUC and pBI221 prepared by modifying pBI121 (manufactured by Clontech) and pBI221 (manufactured by Clontech), which are often used as plant transformation gene vectors.
Using -LUC + (Kazuyuki Hiratsuka Cell Engineering Separate Volume, Plant Cell Engineering Series 6 (1997)), a plasmid vector created by inserting the DNA fragment was constructed. FIG. 3 shows an outline of the transformation vector used. 1
The structure of 21-LUC is shown above and the structure of 221-LUC + is shown below. In constructing the plasmid for meiosis-specific expression, the nucleotide sequences shown in SEQ ID NO: 1 in the sequence listing and SEQ ID NO: 2 in the sequence listing were inserted instead of CaMV 35S pro shown in FIG. Major restriction enzyme cleavage site and NOS
The locations of the terminator (NOS-ter) and the kanamycin resistance gene (NOTII) are indicated.

(Introduction and Transformation of Vector) Agrobacterium tume
facilens LBA4404) by electroporation. Using it, tobacco was transformed by the leaf disk method to obtain a plant. Seeds obtained from the transformed plant were transformed into MS containing kanamycin.
Seed on an agar medium and leave the kanamycin-resistant plants as they are,
Alternatively, they were transplanted and grown in culture soil, and used for observation.

(Measurement of Luciferase Activity) Luciferase activity, which is an indicator of transcriptional activation, was measured for the transformed plants. When the DNA fragment of the present invention was used as a promoter, the luciferase activity was particularly high in anthers containing many meiotic cells, and the luciferase activity was extremely low in other organs and tissues. That is, anthers and leaves, stems, which contain many cells in meiosis,
Tissues such as roots were separated, and extraction was performed according to the attached means using an extraction buffer attached to a commercially available luciferase activity measurement kit, and photon counting was performed for 10 seconds using a Luminometer LB9501 manufactured by Berthold Co., Ltd. to emit light. The activities were compared. FIG. 4 shows the results of measuring the luciferase activity of each tissue by introducing the promoter of the nucleotide sequence shown in SEQ ID NO: 1 in the sequence listing, and FIG. 5 shows the results obtained by introducing the promoter of the nucleotide sequence shown in SEQ ID NO: 2 in the sequence listing. FIG. 6 shows the results obtained by measuring the luciferase activity of each tissue.
In addition, cauliflower mosaic virus 35
The results of measuring the luciferase activity of each tissue by introducing the S promoter are shown below (vertical axis: count /
sec / tube).

As a result, the activity was extremely low in leaves, stems and roots, and was lower than the detection limit, regardless of whether the nucleotide sequence of SEQ ID NO: 1 or the promoter of SEQ ID NO: 2 was introduced. In most cases, the activity was several hundred times higher than that in anthers (FIGS. 4 and 5). This indicates that the DNA fragment has an effect of specifically causing transcriptional activation in meiotic anthers. Also, leaves other than anthers,
Since no transcriptional activation is observed in stems and roots, the temporal and spatial control provided by the DNA fragment is extremely strict and clearly shows that its specificity is extremely high. . When the transgenic tobacco using the cauliflower mosaic virus 35S promoter was used as a control, extremely high transcriptional activation was observed in leaves, stems and roots, but in anthers, the transcriptional activation was lower than those in tissues. It showed activity (FIG. 6).

(Expression of GUS gene)
In order to examine the action of the A fragment as a promoter in more detail, transgenic tobacco was prepared using the GUS gene instead of the luciferase gene, and the state of expression of the GUS gene was examined. For detection of GUS gene expression, X
-Histochemical staining using Gluc as a substrate (Jefferson et al. EMBO J. 6:
3901-3907 (1987)) (FIG. 7).
As a result, blue-green pigment deposition was observed in the meiotic pollen mother cells. This confirmed that the specific expression in anthers indicated by the luciferase activity was derived from the specific expression in pollen mother cells.

From the above results, the plasmid vector DN for plant transformation was constructed using the DNA fragment as a promoter.
It has been shown that insertion into A makes it possible to impart meiosis-specific transcriptional activity, that is, it has an effect of bringing about meiosis-specific gene expression.

[0022]

According to the present invention, the nucleotide sequence of a promoter that specifically acts in meiotic cells has been provided.

[0023]

[Sequence List] <110> Applicant's name: President of Nara Institute of Science and Technology <120> Title of the invention: Plant promoter specific to meiotic cells <160> Number of sequences: 2 <210> SEQ ID NO: 1 < 211> Length of sequence: 1309 <212> Sequence type: Nucleic acid <213> Origin: Teppo lily pollen mother cell <400> Sequence ATGCATGCAC ATGTAAGTCC TGCGTACTCA TATAAAAAAG TGAACATACA CACGAGTATG 60 GTTTGCTTCC ACCTAGTCAT CGTATATTTC ATCACTACTGATCACGATATTCATTCATCATTACGATATTACTGATATCGACATATCGAC ACATAATAAT GTCAACCTTC ACTTCAACTT ACAACCCATG TATGCATACT TTTTTTATCC 240 ACTACATGGA CCTATATATA TAGAAATTTG AACTAAATAT CCAGCTTCAT TAATATTGTT 300 AGCTTAACAT GCATCTCTAC TTCATTATGC ATCTTGATGT TCCCATTGAT GAAATCTTCC 360 TCACTAATAA TCATCACACA TCTAGCCTCT TCACCAGAGA GCATCACACA TCAACAGTCT 420 TCAAAATTCA GTGTCCAACA ACACGGATCT TATTGTGCAT CAGTTAACAC CCTGCGTACC 480 CACTTCACCA AAGATCATAA CACAT TATTG GCCTTCAAAA TTCAATGCCC AACAACACGA 540 ATCATGTCGT ACATCAATCA TGTCGTACAT CAGTTATCAC CCCGCGTTCT AGCTGCATCT 600 TCCAAAGGCT GCTTACGCAC CCCGCAATGG TAGAATGATC AAGCTGGTTG TTCCTATAAG 660 TGAGTAGAGA AACACATGAA TCAGGCTCCA CAAACACATA CTGTATTTCT TTTCTTAACG 720 TCATTTTTAA CAAATTATAA TATTAAAATT TCTCTAAGGT CAGAGATTAC CACACATAAA 780 AAAGTAAATT TTAGTATCTT TAAACTCATA TAAAATAGAA TGGACAAAAT ATTATAAATC 840 ATAAGACAAC CAACCAATTC CACATTATAC TAAATGAAAT ATCACAAATA TGACAAATAA 900 TTAAAAAAAA AAATCGCATG GTTTTCAGAA ATTGGAGGGG TAAGATAAGA AATAAGTCAT 960 CCTCAAAATA CGCGGCGGGG CACGTGTGAA TTCATTTTGT CGGTAGAAAC ACAGCTCGTT 1020 TTTTACTATG ACGATACCAA CCTTACGTAA TACATTCGTT CCATCAACAT GATCGGAATT 1080 GACACAGAAA ACCTCGGTTC AAATTTCACC GGTTTCAATC TCGACCGCTA GCCGGCCGAA 1140 GGCAGACCTC TGCCACCCAT GAAAGCACGA GAAGCTTCCA TCCTCTTCTT GACCCTTTGT 1200 GAAGCATCGA GACCTCTTCC CTCAGTCTTT ATATAAACCC CCAACCCACT CCCAATTCCA 1260 CCGTCTCCAT CGAACACCAG ATAAGTGGAC CTCAAAGAAA TCAAGATCC 1309 <210> SEQ ID NO: : 2 <211> distribution Length: 1461 <212> sequence type: nucleic acid <213> Origin: lily pollen mother cell <400> sequence ATGTACGAAC TATTGCACAC TCTTGAGGGC TCCCTACGCT GCCTAAGCTA TGTGCTTTCA 60 CCTTCTCTAC TCGACATGCA CTCTCACATT GGAGGGGTGT CGCAGGGCTC ATACTCTGCG 120 CTCCTTCTCA AAAGGTTTTC TCCCTCACAG GTTTCTCTCC ACGTCAACCC AAGCTCAAAA 180 ATTCGGGGCC CAGCCCGCTG AGGATATTTT TGGCGTCATT AGTTGTTGAT GATGAATTAT 240 TCATTCCTAA AATCTTGTCA TGAATGCTTG ATTTTTTCAT CATTTATCTT ATGTATTTCC 300 TCCAAACTTG TATTGCTTAT CATGCACAAA AGAGTTTAAC TTACATTAGT GACATCAAGC 360 AAATACATGG AATACACGTT AACAAATGAT TAAATGTTGG TTATAATTGT GCACAATTAT 420 TGTCTGGTCA CCTAGTCATC TTCTTTCATG AATCCTCCAC TGATGAGATC CTCCCTCAAT 480 GAGTTCATTC TCAAACAAGT TACCATTACC CGTAGAAGGA AAAGAGGAAG AAAAATAAAC 540 ACGTGATGAT GACCAAAAGT AATGAAAGAG AGAACACGAT AAATGATGTT CGAGAAAACA 600 ATCCCAAAAT ATATGGATAC GATATTGAAA TACTTATTGA GAAACCCAAA CAAAACAAGC 660 ATGCACTGAC ATAGGAATAT TGTGACTTTT GTGTGCCCTT GCAAGGTGAT TGACGATAGA 720 TCATAATCAA CATCCT TACA CTTTACCGTT CGCCCACCAT CCCTTTGTTC CTCCTTCCCT 780 CTTTCGAAGA TGTTTTCTCG TTATCTTTGC TACTTTCCTC ATAAATAGAC ATCGTATCAT 840 TTTTGGCGTA AATCATTTTT TTGTTGTTGA ATTTTAGCGA AATTCACCCA GACCATCCAT 900 GTGCACCACG ACAAATTTAT CCCCGAGACA TTGTGGCATA AAGTTACTAA TGACTTGATA 960 GCCTAGTGGT TGGGACTTGG ATGGTTATCA ATGAGGTCTG TGGTTCGAGT CTCGGAAAAC 1020 TACTGGAAAC AGCCTCCATT TAATAAATAG GAATGCTGCG TACATTTCTG TCCCCCGTAA 1080 GCCTGCGCAA GCGAGATAGG AATGCTGCGT ACATTTCTGT CCCCCGTAAG CCTGCGCAAG 1140 CGAGAAAATA CGGATTATCC ACATTGGGGC GTAAAATTAA TCCCCTACAT TTCTGTCCCC 1200 CGTAAGCCTG CACAAGCGAG AAAATACGGA TTATCCACAT TGGGGTGTAA AATTAATCCC 1260 CTACCAACAT GTTTTGTGTC AGTCTGCCTA ACTGGGCCAA TAATAACCCT GCAATCCTCT 1320 GTTAAACTGG AAAATTAATC CGTAGAGTCC CCATCCCGGT TAGAGAACGA AAACTATAAC 1380 TGCTGCAAAG GGCTACTGGG CCCACCAATT TTCACCCGTC TCTGCCCCCC ATTCAGCTAA 1440 CCCAGCTCGC AACGGAGAAC C 1461

[Brief description of the drawings]

FIG. 1 shows the nucleotide sequence of a meiosis-specific promoter derived from LIM10.

FIG. 2 shows the nucleotide sequence of a meiosis-specific promoter derived from LIM18.

FIG. 3 shows a vector for plant transformation, 1
It is a figure which shows the structure of 21-LUC and 221-LUC +.

FIG. 4 is a diagram showing the results of measuring the luciferase activity of each tissue by introducing the promoter of FIG.

FIG. 5 is a diagram showing the results of measuring the luciferase activity of each tissue obtained by introducing the promoter of FIG. 2;

FIG. 6 shows cauliflower mosaic virus 35
FIG. 2 is a diagram showing luciferase activity of each tissue by introducing an S promoter.

FIG. 7 is a photograph in which the specificity of promoter expression was detected by a tissue staining chemical GUS staining method using X-Gluc as a substrate.

──────────────────────────────────────────────────続 き Continued on the front page (72) Inventor Masayoshi Minami 8916-5 Takayamacho, Ikoma City, Nara Prefecture Graduate School Dormitory 1-121 (56) Reference DNA Res. (1994) Vol. 1, p. 15-26 Plant Cell Physiol. (1998) Vol. 39, suppp l. , P. s45 (58) Field surveyed (Int. Cl. ⁷ , DB name) C12N 15/00-15/90 BIOSIS (DIALOG) WPI (DIALOG) GenBank / EMBL / DDBJ

Claims (6)

(57) [Claims] The present invention relates to a meiotic cell.
Base number as described in SEQ ID NO: 1 in the sequence listing
Characterized by comprising a base sequence represented by 1-1309
You, a plant promoter. 2. The method of claim 1 , wherein the promoter is specifically promoted in meiotic cells.
Base number described in SEQ ID NO: 2 in the sequence listing
It is characterized by comprising a base sequence represented by 1-1461.
You, a plant promoter. 3. An expression vector into which the plant promoter according to claim 1 has been inserted. 4. The plant promoter according to claim 1 or 2 is inserted into an expression vector, the expression vector is introduced into a plant cell to obtain a transformed cell, and a plant seed is obtained from the transformed cell. A method for producing a transformed plant, comprising a step of producing a transformed plant from plant seeds. 5. A transformed plant into which the plant promoter according to claim 1 or 2 has been inserted. 6. A plant seed derived from the transformed plant according to claim 5.