JP2002272472A - Gene encoding sulfate ion transporter Sultr1; 3 - Google Patents

Abstract

[PROBLEMS] To provide a novel sulfate ion transporter gene derived from Arabidopsis thaliana and a transformed plant into which this gene has been introduced. SOLUTION: Arabidopsis thaliana (Arabidopsis thalian)
a) a gene which is present on chromosome 1 of a) and encodes a sulfate ion transporter Sultr1; 3 having a specific amino acid sequence, and a transformed plant into which this gene has been introduced

Description

DETAILED DESCRIPTION OF THE INVENTION

[0001]

TECHNICAL FIELD The invention of this application relates to a novel sulfate ion transporter Sultr1; 3 derived from Arabidopsis thaliana.
And a transgenic plant into which this gene has been introduced.

[0002]

2. Description of the Related Art Sulfur is a basic nutrient in plants. To synthesize sulfur-containing organic compounds from sulfur,
Uptake of sulfate ions from roots is important (Plant Ph
ysiol. 120: 637-643, 1999). Physiological studies suggest that uptake and translocation of sulfate involve a few sulfate translocation systems with different affinities for sulfate (Plant Phys.
iol. Biochem. 34: 409-416, 1996 etc.). In fact, the inventors of the present application have so far isolated the seven sulfate ion transporter genes shown in Table 1 and reported their structures and functions (FEBS Lett. 392: 95-99, 1996). ; Proc.
Natl. Acad. Sci. USA, 94: 11102-11107, 1997; J. Ex.
p. Bot. 50: 1713-1714, 1999; Plant Physiol. 121: 6
86, 1999).

[0003]

[Table 1]

Further, the inventors of the present application have reported that these sulfate ion transporters are each predominantly expressed in different organs of a plant individual. For example, Sultr1; 1 is expressed in the lateral root cap, root hair, epidermis and cortex, and Sultr2; 1 is expressed in the root column, leaf xylem parenchyma cells and leaf phloem cells (Plant J. 23: 171-182, 2000).
In addition, since Sultr1; 1 and yeast into which Sultr2; 1 gene has been introduced grow well in a medium using sulfate ions as a sulfur source,
It has also been confirmed that expression of these genes is closely related to sulfate ion uptake and translocation (Plant J.
Id.).

[0005]

As described above, the sulfate ion transporter gene is a plant gene involved in the uptake and translocation of succinate ion which plays an important role in plant sulfur absorption. It is expected that the growth and yield of agricultural products and the like can be significantly improved by transducing other plants and protein coding regions into high expression of sulfate ion transporters. In addition, the introduction of this gene makes it possible to improve the variety of crops that are difficult to grow on soil with a low sulfate ion concentration due to poor sulfur use efficiency. Furthermore, the introduction of this gene enhances the ability of plants to absorb sulfur, thereby improving the production of sulfur-containing metabolites such as glutathione and phytokeratin, thereby producing plants with enhanced antioxidant and heavy metal tolerance. In addition, it is possible to obtain a large amount of a sulfur-containing metabolite having an antioxidant action and a heavy metal resistance action from these plants.

[0006] Thus, introduction of a sulfate ion transporter gene into a plant is expected to have many excellent effects. However, as described above, each of the sulfate ion transporter genes is associated with a plant individual. In order to produce a transgenic plant for any purpose, it is necessary to prepare as many sulfate ion transporter genes as possible and to introduce genes according to their characteristics in order to produce transformed plants for various purposes. is necessary.

The invention of this application has been made in view of the above circumstances, and has as its object to provide a new sulfate ion transporter gene. Another object of the invention of this application is to provide a transformed plant individual into which the novel gene has been introduced.

[0008]

SUMMARY OF THE INVENTION The present application discloses an Arabidopsis thaliana (Arabid) as an invention for solving the above-mentioned problems.
opsis thaliana) on SEQ ID NO: 2
Sulphate transporter Su with the amino acid sequence of
Provide the gene encoding ltr1; 3.

[0009] More specifically, this gene has its cD
NA is a gene having the nucleotide sequence of SEQ ID NO: 1. The application also provides a polynucleotide purified from the genomic DNA, mRNA, cDNA, or the complement thereof, of the gene.

[0010] This application also provides an oligonucleotide consisting of a base sequence of 10 base pairs or more, which hybridizes to the gene or polynucleotide. Furthermore, this application provides a recombinant vector carrying the polynucleotide.

This application further provides a plant into which the above-mentioned gene has been introduced, or a progeny thereof or a tissue thereof. That is, the inventors of the present application conducted a search of the Arabidopsis thaliana genomic sequence based on the known sulfate ion transporter gene cDNA bases shown in Table 1, and revealed the existence of a novel gene having high homology. did. Based on the genomic sequence, set the synthetic primers at both ends of the coding region which is expected to give a cDNA fragment by RT-PCR method. The promoter region was isolated by PCR using genomic DNA as a template. Furthermore, the present invention was completed by confirming the characteristics (function and locality of expression).

The novel sulfate transporter gene Sultr1; 3 encodes a sulfate transporter protein having a full-length cDNA of 1971 bp (SEQ ID NO: 1) and a sequence of 656 amino acid residues (SEQ ID NO: 2). I have.
The Sultr1; 3 gene is specifically expressed in the phloem of the root.

The invention of this application is based on the novel sulfate ion transporter gene as described above. In the present invention, `` polynucleotide '' and `` oligonucleotide '' do not mean a fragment having a specific number of bases. There are exceptions.

Hereinafter, embodiments of the present invention will be described in detail.

[0015]

BEST MODE FOR CARRYING OUT THE INVENTION As described above, the gene Sultr1; 3 of the present invention is present on chromosome 1 of Arabidopsis thaliana,
This is an Arabidopsis thaliana genome gene encoding the sulfate ion transporter Sultr1; 3 having the amino acid sequence of SEQ ID NO: 2. More specifically, this Sultr1; 3 gene
It is characterized in that the cDNA has the nucleotide sequence of SEQ ID NO: 1.

The Sultr1; 3 gene can be used for producing a polynucleotide and also for producing a transformed plant. The Sultr1; 3 gene also contains control regions (promoter / enhancer, suppressor, etc.) for expression of the protein encoded by the gene. These expression control regions contain the protein Sultr1; 3
It is useful for searching for the function of Sultr1; 3 or the expression regulator of the Sultr1; 3 gene. Such an expression control region can also be used for specifically controlling the expression of another gene in the root phloem.

The Sultr1; 3 gene of the present invention can be obtained, for example, by using a polynucleotide or an oligonucleotide consisting of the nucleotide sequence of SEQ ID NO: 1 or a partial sequence thereof as a probe under the stringent conditions and the genome of Arabidopsis thaliana.
It can be isolated by screening a DNA library.

[0018] The polynucleotide of the present invention includes genomic DNA and mRNA isolated from Arabidopsis thaliana cells (particularly roots).
DNA fragments and R
It can be obtained as an NA fragment. In addition, cDNA is used as a template by using poly (A) + RNA extracted from Arabidopsis thaliana as a template (Mol. Cell Biol. 2, 161-170, 1982; J. Gen.
e 25, 263-269, 1983; Gene, 150, 243-250, 1994). Alternatively, the target cDNA can also be synthesized using an oligonucleotide as a primer and an RT-PCR method using mRNA isolated from Arabidopsis thaliana as a template. CDNA prepared in this way
Has, specifically, the base sequence of SEQ ID NO: 1. In general, polymorphisms due to individual differences are frequently observed in eukaryotic genes. Therefore, in the nucleotide sequence of SEQ ID NO: 1,
Polynucleotides in which one or more (10 or less) nucleotides have been added, deleted and / or substituted with other nucleotides are also included in the scope of the polynucleotide (cDNA) of the present invention.

These polynucleotides can also be used for producing transformed plants. The oligonucleotide of the present invention is an oligonucleotide consisting of a base sequence of 10 base pairs or more that hybridizes to the Sultr1; 3 gene or its polynucleotide under stringent conditions (a DNA fragment or an RNA fragment, or an antisense strand thereof) ). This oligonucleotide is useful, for example, as a probe for isolating the Sultr1; 3 gene or as a PCR primer for synthesizing a polynucleotide. Such an oligonucleotide can be synthesized and prepared by a conventional method, or in some cases, can also be prepared by cleaving the polynucleotide of the present invention with an appropriate restriction enzyme.

As the recombinant vector of the present invention, an appropriate one is used depending on the kind of the polynucleotide as the insert and the purpose of use. For example, cDNA or its
When the ORF region is used as an insert for plant transformation, a plasmid vector or the like provided with a promoter sequence or the like that works in plants is used. When genomic DNA of Sultr1; 3 gene is used as an insert, BAC (Bact
(Erial Artificial Chromosome) vector and cosmid vector can also be used.

The transgenic plant of the present invention is
ltr1; 3 gene transduced, higher Sultr than wild type
It is a transgenic plant characterized by having 1; 3 activity. This transformed plant is high
The Sultr1; 3 activity has excellent sulfate ion absorption and transfer efficiency and sulfur can be used efficiently, so it is excellent in growth and product yield. In addition, it can be satisfactorily grown even in soil having a low sulfate ion concentration. Furthermore, due to its high sulfur-absorbing ability, it produces many sulfur-containing metabolites such as glutathione and phytokeratin, which binds to heavy metals such as cadmium to detoxify heavy metals, or obtain antioxidant ability through sulfur-containing metabolites I do.

The transformed plant of the present invention is, for example, Sult
The r1; 3 polynucleotide (cDNA) can be converted to a promoter that functions in plants (for example, cauliflower mosaic virus (CaM
V) 35S promoter, etc.), recombination into a vector, and transduction of this vector into plants.

The plant into which the polynucleotide is introduced is not particularly limited. It may be annual or perennial. Alternatively, it may be an aquatic plant. However,
Plant species that do not affect the growth and the like of other useful plant species by sowing their seeds and the like and breeding on agricultural land and the like, such as so-called weeds, are preferred. Further, it is particularly preferable that the plant is a useful plant species. That is, food crops (rice such as rice, upland rice, barley, wheat and other wheat, corn, buckwheat, soybean, adzuki, sweet potato, potato, etc.), horticultural crops (apple, pear, loquat, peach, plum, cherry, grape, Deciduous fruit trees such as figs and oysters; citrus fruits such as mandarin, lime, lemon, yuzu and kumquat; tropical fruit trees such as coconut, pineapple, banana, mango, papaya; cucumber, watermelon, pumpkin and other cucumber vegetables; eggplant and tomato Vegetables vegetables such as peas, kidney beans, broad beans; Fruit vegetables such as strawberries and okra; Direct root vegetables such as radish and carrot; Tuberous vegetables such as taro and ginger; Leek and garlic Green onion vegetables; vegetable vegetables such as cabbage and Chinese cabbage; raw and spicy vegetables such as celery, lettuce and udo; soft vegetables such as honeybee, seri, butterbur Vegetables and vegetables; wild vegetables; morning glory, salvia sweet pea, etc., two-year herbaceous plants; chrysanthemums, carnations, etc .; bulbous flowers, such as crocuses and tulips; orchids; , Azaleas, cherry blossoms, etc .; branch trees such as willow, eucalyptus, etc .; landscaping trees, such as dogwood, nanten etc .; Sugar crops such as cotton, thyme, rush sugar; starch crops such as konjac, jerusalem artichoke; medicinal crops such as jojoba, peppermint, poppy; tea, tobacco,
Hop crops such as hops; paper crops such as cucumber, honey beetle; dye crops such as eye and safflower; spice crops; fragrant crops; rubber and resin crops such as para rubber tree and rush; and fertilizer crops (timothy, orchard grass) , Ryegrass, fescue, bentgrass, bromegrass, sorghum, rosegrass, bermudagrass,
Grass feed crops such as oats; legume feed crops such as clovers, alfalfa, and vetch; feed leaves and root crops such as feed turnips and livestock peat; fertilizer trees; and green manure crops such as vetch. can do.

Transformation of the plant is carried out by appropriately employing a known method (leaf disk co-culture method, electroporation method, Agrobacterium method, particle gun method, etc.) according to the type of the plant and the like. Can be. For example, the rice field uses electroporation (Nature 3
38: 274, 1989), Agrobacterium method (Plant J. 6: 2
71, 1994) or the particle gun method (Plant Cell Re
14: 586, 1995). For example, in the case of the electroporation method, a recombinant vector based on a sulfate ion transporter gene is introduced into rice protoplast cells by applying an electric pulse, and the transformed protoplasts are cultured to obtain calli. By culturing the callus, a transformed rice individual (primary) into which the sulfate ion transporter gene has been introduced can be obtained.

When rapeseed and tobacco are transformed by the Agrobacterium method (Plant Mol. Biol. 26: 1115, 1994), the dried seeds are cultured for two to three weeks and germinated and grown aseptic. The hypocotyl is cut to an appropriate length, placed on a preculture medium, and precultured in the light overnight. Next, the previously hypocultured hypocotyl was put into a suspension of Agrobacterium having a recombinant plasmid with a sulfate ion transporter gene, and cultured.The solution was filtered, and only the hypocotyl was removed. Agrobacterium is removed and co-cultured on the original preculture medium for two nights to infect the hypocotyl with Agrobacterium. Thereafter, the hypocotyl was transferred to a sterilization medium and cultured for about 3 days to suppress the growth of Agrobacterium. Then, these hypocotyls were cultured in a selection medium to obtain a transformed callus. Germination medium, part
After culturing and acclimating in an elongation medium and growing in an isolated greenhouse, transformed tobacco and rape (primary) can be obtained. Also, seeds can be obtained by self-pollinating this primary plant.

Furthermore, when the transformation is performed by using the leaf disk co-culture method using Agrobacterium (EMBO J. 6: 2531, 1987), sulfate ions are added to tobacco leaves cut into pieces of about 1 cm square. A suspension of Agrobacterium harboring the recombinant plasmid with the transporter gene is added, co-cultured for about 2 days, and the section (leaf disk) is washed and placed on a medium. Then, by selecting a transformant and continuing the culture, callus formed at the cut end of the leaf vein of the leaf disk covers the entire surface of the leaf,
A shoot is then formed from the callus. Green shoot
Is excised and cultured for rooting to obtain a regenerated transformant.

When tobacco or rape is transformed by electroporation (Plant Tissue Culture Letters 11: 199, 1994), protoplasts are obtained from the hypocotyls that have been germinated aseptically, and an electric pulse is applied to the protoplasts to apply sulfate to the protoplasts. A recombinant plasmid carrying the ion transporter gene is introduced. Transformed cells are selected, and the resulting transformed callus is further cultured to obtain a regenerated individual (primary). The acclimated plants can be grown in an isolated greenhouse and self-pollinated to obtain seeds.

The transformation of each of the other plants can be performed by a known method. For example, the Agrobacterium method (Bio / tech
nology 6: 915, 1988), electroporation method (Plant Cell Rep. 10: 106, 1991), particle gun method (Theor. Appl. Gnet. 79: 337, 1990), and the like.

Hereinafter, the invention of this application will be described in more detail and specifically with reference to examples, but the invention of this application is not limited to the following examples.

[0030]

EXAMPLES Example 1 Cloning of cDNA The cDNA of the Sultr1; 3 gene of the present invention was cloned by the following method.

A search was made for the genomic sequence of Arabidopsis thaliana based on the nucleotide sequence of the translation region of the gene Sultr1; 1, which encodes the high-affinity sulfate ion transporter of Arabidopsis thaliana. There were genes showing homology. Therefore, this gene was named Sultr1; 3, and the translation region was isolated. 1st strand cDNA was synthesized using total RNA extracted from the roots of Arabidopsis thaliana (A. thaliana ecotype Columbia) grown on an MS-modified medium containing 0.1 mM sulfate ion as a sulfur source and germinated for 2 weeks after germination. Synthetic primers 1; 3FEc (SEQ ID NO: 3) and 1; 3R having EcoRI sites at both ends of the predicted translation region
Ec (SEQ ID NO: 4) was set, and PCR was performed using the 1st strand cDNA as a template. The amplified fragment was cloned into pBluescriptII SK- (Stratagene) using the constructed EcoRI site. The nucleotide sequence was determined, and it was confirmed that the Sultr1; 3 cDNA clone was isolated. Example 2: Complementation test of yeast sulfate transporter deficient mutants Yeast sulfate transporter genes SUL1 and SU
The yeast mutant CP154-7A lacking L2 was replaced with the Su obtained in Example 1.
By introducing the cDNA of the ltr1; 3 gene, the expression product of Sultr1; 3 was constantly expressed. Also, Sultr1; 1, Sultr1; 2, Sult
The expression products of r2; 1 and Sultr2; 2 were expressed. Specifically, according to Example 1, Sultr1; 3 cDNA previously cloned into pBluescriptIISK- was cut at the EcoRI site, and pYE22m
(Appl. Mocrobiol. Biotechnol. 30: 515, 1989)
It was inserted into the oRI site in the sense direction. The constructed plasmid and pYE22m were subjected to the lithium acetate method (Nuc. Acids. Res.
20: 1425, 1992). Complementation experiments included SD (Methods Enzymol. 247: 709, 1).
An SD (ΔS) -modified medium (Gen. Genet. 247: 709, 1995) in which the sulfate salt of 995) was replaced with a chloride salt was used.

The results are as shown in FIG. 1. Although the yeast into which only the expression vector pYE22m was introduced cannot grow on a medium containing 0.1 mM sulfate ion as a sulfur source, the yeast expressing Sultr1; 3 did not. It grew well. The Su of the present invention
Yeast expressing ltr1; 3 grew with equal or higher efficiency than yeast expressing other sulfate ion transporter genes. Example 3 Preparation of Transformed Arabidopsis thaliana The promoter region of Sultr1; 3 gene was isolated by the following method. Based on the Arabidopsis genome sequence, XhoI
Synthetic primer 1 with site; 3G-FXh (SEQ ID NO: 5)
And 3G-RNc (SEQ ID NO: 6) were synthesized, and PCR was performed using these primers.
The 5'-region of 1; 3 was amplified. The nucleotide sequence of the obtained amplified fragment was determined, and it was confirmed that the DNA fragment containing the promoter region of Sultr1; 3 was isolated.

Next, the GFP expression vector pTH-2 (Curr. Bi
ol. 6: 325, 1996), the NcoI-NotI fragment containing the GFP coding region was fused to the promoter fragment to construct a promoter-GFP fusion gene. The XhoI-NotI fragment of this promoter-GFP fusion gene was inserted into the XhoI-NotI site of pBluescriptII SK-, and a SacI site was added downstream of the NotI site. The promoter-GFP fusion gene XhoI-SacI fragment was cut out from the obtained plasmid, and a binary vector pBI
The recombinant vector BIN was replaced with the SalI-SacI region containing the translation region of the β-Glucuronidase gene in 101 (Clontech).
1; 3-GFP was constructed.

[0034] The recombinant vector BIN1; 3-GFP was transformed into an planta transformation method using Agrobacterium (Plant Phy
siol. 124: 1540, 2000 etc.) to introduce Arabidopsis thaliana to produce transformed Arabidopsis thaliana. In particular,
A. tumefaciens C58C1GV3101 (pMP90) (Mol. G) was prepared by electroporation of the constructed vector BIN1; 3-GFP.
en. Genet. 204: 383, 1986). In planta transformation of Arabidopsis thaliana, the floral dip method (Plant J. 1
6: 735, 1998). The obtained T1 seeds were aseptically sterilized with a kanamycin 50 mg / l GM agar medium (Pro.
cad. Sci. USA. 85: 5536, 1998), and transformants showing kanamycin resistance were selected.

FIG. 2A is a micrograph of the roots of transformed Arabidopsis thaliana, and FIG. 2B is a photographic image (left) and a fluorescent photographic image (right) of a transverse section of the root. As is clear from FIG. 2, GFP expressed by the Sultr1; 3 promoter was:
Expression was observed only in the root phloem.

[0036]

As described above in detail, the present application provides a new sulfate ion transporter gene derived from Arabidopsis thaliana and a transformed plant into which this gene has been introduced. This transformed plant is excellent in sulfate ion absorption and transfer efficiency due to high Sultr1; 3 activity and can use sulfur efficiently, so that it is excellent in growth and product yield. Further, it can be satisfactorily grown even in soil having a low sulfate ion concentration. Furthermore, due to its high sulfur-absorbing ability, it produces many sulfur-containing metabolites such as glutathione and phytokeratin, which binds to heavy metals such as cadmium to detoxify heavy metals, or obtain antioxidant ability through sulfur-containing metabolites I do. Furthermore, by using this transformed plant, it becomes possible to produce useful sulfur-containing metabolites such as glutathione and phytokeratin in large quantities.

[0037]

[Sequence List] SEQUENCE LISTING <110> Japan Science and Technology Corporation <120> A gene encoding sulfate transporter 1; 3 <130> NP01035-YS <140> <141> <160> 2 <170> PatentIn Ver. 2.1 <210 > 1 <211> 1971 <212> DNA <213> Arabidopsis thaliana <220> <221> CDS <222> (1) .. (1971) <300> <308> GenBank / AB049624 <309> 2000-10-07 <400> 1 atg tcg gct aga gct cat cct gtg gac gat gac gga gag att tcc ccg 48 Met Ser Ala Arg Ala His Pro Val Asp Asp Asp Gly Glu Glu Ile Ser Pro 1 5 10 15 gtg gaa aga tct tcc ccg cgg caa gca aat aca ccg tat gtc cac aaa 96 Val Glu Arg Ser Ser Pro Arg Gln Ala Asn Thr Pro Tyr Val His Lys 20 25 30 gtc gaa gtt cct cct aag caa aac ctt ttc aat gag ttc atg tac act 144 Val Glu Val Pro Pro Lys Gln Asn Leu Phe Asn Glu Phe Met Tyr Thr 35 40 45 ttt aaa gaa act ttc ttc cac gat gat cct cta agg cat ttc aag gac 192 Phe Lys Glu Thr Phe Phe His Asp Asp Pro Leu Arg His Phe Lys Asp 50 55 60 cag tca aaa tcc aaa aag ctc atg ctc ggt atc cag tcc gtc ttt ccg 240 Gln Ser Lys Ser Lys Lys Leu Met Leu Gl y Ile Gln Ser Val Phe Pro 65 70 75 80 gtt atc gag tgg gga aga aaa tat aat ctt aag ttg ttt cgc ggc gat 288 Val Ile Glu Trp Gly Arg Lys Tyr Asn Leu Lys Leu Phe Arg Gly Asp 85 90 95 ctt att gcc ggt tta acc ata gcc agt ctc tgc att cct cag gat att 336 Leu Ile Ala Gly Leu Thr Ile Ala Ser Leu Cys Ile Pro Gln Asp Ile 100 105 110 gga tat gca aag ctc gcg agt ctt gac cct aag tat ggt cta tat tca 384 Gly Tyr Ala Lys Leu Ala Ser Leu Asp Pro Lys Tyr Gly Leu Tyr Ser 115 120 125 agt ttt gtt cct ccg ttg gtg tac gca tgt atg gga agc tca aag gat 432 Ser Phe Val Pro Pro Leu Val Tyr Ala Cys Met Gly Ser Ser Lys Asp 130 135 140 ata gcg att gga ccc gtt gca gtg gtt tca ctc cta tta ggt act ctg 480 Ile Ala Ile Gly Pro Val Ala Val Val Ser Leu Leu Leu Gly Thr Leu 145 150 155 160 ctt cgt gct gag atc gac ccc aac aca aac cct aat gaa tat ctc cgc 528 Leu Arg Ala Glu Ile Asp Pro Asn Thr Asn Pro Asn Glu Tyr Leu Arg 165 170 175 tta gcc ttc acc tcc acg ttc ttt gcc ggt gtc act caa gca gca ctc 576 Leu Ala Phe Thr Thr Phe Phe Ala Gly Val Thr Gln Ala Ala Leu 180 185 190 gga ttc ttc aga ttg ggt ttc ttg atc gat ttc ttg tcc cac gcg gcg 624 Gly Phe Phe Arg Leu Gly Phe Leu Ile Asp Phe Leu Ser His Ala Ala gta 200 205 ggt ttc atg ggc gga gca gcc atc acc att gcg ctg caa cag 672 Val Val Gly Phe Met Gly Gly Ala Ala Ile Thr Ile Ala Leu Gln Gln 210 215 220 ctc aaa ggc ttc cta gga atc aat aag ttc aca atag aaa acc 720 Leu Lys Gly Phe Leu Gly Ile Asn Lys Phe Thr Lys Lys Thr Asp Ile 225 230 235 240 atc gcg gtt ctt tct tcc gta atc agc tca gcc cat cac gga tgg aat 768 Ile Ala Val Leu Ser Ser Val Ile Ser Ser Ala His His Gly Trp Asn 245 250 255 tgg cag aca ata ctc att agt gca tcg ttc ttg atc ttc ctt ctc atc 816 Trp Gln Thr Ile Leu Ile Ser Ala Ser Phe Leu Ile Phe Leu Leu Ile 260 265 270 270 tcc aag ttt atc ggg aag aac aag aaa ctg ttt tgg att cca gct 864 Ser Lys Phe Ile Gly Lys Arg Asn Lys Lys Leu Phe Trp Ile Pro Ala 275 280 285 att gct ccg tta gta tct gtc atc att tca acc ttc ttc gtc tac ata Pro 912 AleVal Ser Val Ile Ile Ser Thr Phe Phe Val Tyr Ile 290 295 300 acc cga gcc gac aag aaa gga gtt cag ata gtg aaa cat ctt gac aaa 960 Thr Arg Ala Asp Lys Lys Gly Val Gln Ile Val Lys His Leu Asp Lys 305 310 315 320 ggt ctg aac cct tct tct ttg cgt cta ata tat ttc tcg ggc gat tac 1008 Gly Leu Asn Pro Ser Ser Leu Arg Leu Ile Tyr Phe Ser Gly Asp Tyr 325 330 335 ctt ctc aag ggc ttc cgc ata ggca gtt atg gtt gcc ttg 1056 Leu Leu Lys Gly Phe Arg Ile Gly Val Val Ser Gly Met Val Ala Leu 340 345 350 acg gaa gct gta gcg ata gga aga aca ttt gca gca atg aaa gac tac 1104 Thr Glu Ala Val Ala Ile Gly Arg Thr Phe Ala Ala Met Lys Asp Tyr 355 360 365 caa atc gat ggt aac aaa gag atg gta gca tta gga gca atg aac gta 1152 Gln Ile Asp Gly Asn Lys Glu Met Val Ala Leu Gly Ala Met Asn Val 370 375 380 atc ggt tca atg acc tct tgc tat gta tcc acc ggt tct ttc tca aga 1200 Ile Gly Ser Met Thr Ser Cys Tyr Val Ser Thr Gly Ser Phe Ser Arg 385 390 395 400 tcc gcc gtc aac ttt atg gcc gga tgt caa acg gca gtc tcc aac at c 1248 Ser Ala Val Asn Phe Met Ala Gly Cys Gln Thr Ala Val Ser Asn Ile 405 410 415 atc atg tcc att gtc gtc ctc tta acg ctt ctc ttc ctc act cct ctt 1296 Ile Met Ser Ile Val Val Leu Leu Thr Leu Leu Phe Leu Thr Pro Leu 420 425 430 ttc aaa tac aca ccc aac gca att ctt gca gcg atc atc atc aac gct 1344 Phe Lys Tyr Thr Pro Asn Ala Ile Leu Ala Ala Ile Ile Ile Asn Ala 435 440 445 gtg att cct ttg gtt gac aat gct acc att ttg atc ttc aag atc 1392 Val Ile Pro Leu Val Asp Val Asn Ala Thr Ile Leu Ile Phe Lys Ile 450 455 460 gat aag ctc gat ttt gtt gct tgt atg ggg gct ttt ttt ggt gtc atc 1440 Asp Lys Leu Asp Phe Val Ala Cys Met Gly Ala Phe Phe Gly Val Ile 465 470 475 480 ttt gta tcc gta gag att ggc ctt cta ata gcc gtg ggg ata tct ttt 1488 Phe Val Ser Val Glu Ile Gly Leu Leu Ile Ala Val Gly Ile Ser Phe 485 490 495 gcc aag ata ctt ttg caa gtt acg aga cct agg aca gcg att ctt gga 1536 Ala Lys Ile Leu Leu Gln Val Thr Arg Pro Arg Thr Ala Ile Leu Gly 500 505 510 aag ata cca ggg act tcg gtt tac agg aat atc aat cag tat cct gaa 1584 Lys Ile Pro Gly Thr Ser Val Tyr Arg Asn Ile Asn Gln Tyr Pro Glu 515 520 525 gcg act agg att ccg gga gtt ttg aca att cgt gtt gac tcc gcg att 1632 Ala Thr Arg Ile Pro Gly Val Leu Thr Ile Arg Val Asp Ser Ala Ile 530 535 540 tac ttc tcc aac tcc aat tat gtt agg gaa agg att caa aga tgg ttg 1680 Tyr Phe Ser Asn Ser Asn Tyr Val Arg Glu Arg Ile Gln Arg Trp Leu 545 550 555 560 aca gat gaa gaa gag atg gtg gaa gct gca aga ttg cct agg atc cag 1728 Thr Asp Glu Glu Glu Met Val Glu Ala Ala Arg Leu Pro Arg Ile Gln 565 570 575 ttt ctt atc atc gaa atg tca cct gtt acg gac ggt act 1776 Phe Leu Ile Ile Glu Met Ser Pro Val Thr Asp Ile Asp Thr Ser Gly 580 585 590 att cac gcc tta gaa gac ttg tat aag tct ctc caa aaa cga gac att 1824 Ile His Ala Leu Glu Asp Leu Tyr Lys Ser Leu Gln Lys Arg Asp Ile 595 600 605 cag ttg gtt cta gcg aat cca gga ccg ccg gtc ata aat aag cta cat 1872 Gln Leu Val Leu Ala Asn Pro Gly Pro Pro Val Ile Asn Lys Leu His 610 615 620 gtt tct cac ttt gcg gac ttg ata gga cac gac aaa atc ttt ctg acg 1920 Val Ser His Phe Ala Asp Leu Ile Gly His Asp Lys Ile Phe Leu Thr 625 630 635 640 gtg gct gag gcc gtg gat tct tgc tcc cct aaa ctg tcc gac gag gtc 1968 Val Glu Ala Val Asp Ser Cys Ser Pro Lys Leu Ser Asp Glu Val 645 650 655 tga 1971 <210> 2 <211> 656 <212> PRT <213> Arabidopsis thaliana <400> 2 Met Ser Ala Arg Ala His Pro Val Asp Asp Asp Gly Glu Ile Ser Pro 1 5 10 15 Val Glu Arg Ser Ser Pro Arg Gln Ala Asn Thr Pro Tyr Val His Lys 20 25 30 Val Glu Val Pro Pro Lys Gln Asn Leu Phe Asn Glu Phe Met Tyr Thr 35 40 45 Phe Lys Glu Thr Phe Phe His Asp Asp Pro Leu Arg His Phe Lys Asp 50 55 60 Gln Ser Lys Ser Lys Lys Leu Met Leu Gly Ile Gln Ser Val Phe Pro 65 70 75 80 Val Ile Glu Trp Gly Arg Lys Tyr Asn Leu Lys Leu Phe Arg Gly Asp 85 90 95 Leu Ile Ala Gly Leu Thr Ile Ala Ser Leu Cys Ile Pro Gln Asp Ile 100 105 110 Gly Tyr Ala Lys Leu Ala Ser Leu Asp Pro Lys Tyr Gly Leu Tyr Ser 115 120 125 Ser Phe Val Pro Pro Leu Val Tyr Ala Cys Met Gly Ser Ser Lys Asp 130 135 140 Ile Ala Ile Gly Pro Val Ala Val Val Ser Leu Leu Leu Gly Thr Leu 145 150 155 160 Leu Arg Ala Glu Ile Asp Pro Asn Thr Asn Pro Asn Glu Tyr Leu Arg 165 170 175 Leu Ala Phe Thr Ser Thr Phe Phe Ala Gly Val Thr Gln Ala Ala Leu 180 185 190 Gly Phe Phe Arg Leu Gly Phe Leu Ile Asp Phe Leu Ser His Ala Ala 195 200 205 Val Val Gly Phe Met Gly Gly Ala Ala Ile Thr Ile Ala Leu Gln Gln 210 215 220 Leu Lys Gly Phe Leu Gly Ile Asn Lys Phe Thr Lys Lys Thr Asp Ile 225 230 235 240 Ile Ala Val Leu Ser Ser Val Ile Ser Ser Ala His His Gly Trp Asn 245 250 255 Trp Gln Thr Ile Leu Ile Ser Ala Ser Phe Leu Ile Phe Leu Leu Ile 260 265 270 Ser Lys Phe Ile Gly Lys Arg Asn Lys Lys Leu Phe Trp Ile Pro Ala 275 280 285 Ile Ala Pro Leu Val Ser Val Ile Ile Ser Thr Phe Phe Val Tyr Ile 290 295 300 Thr Arg Ala Asp Lys Lys Gly Val Gln Ile Val Lys His Leu Asp Lys 305 310 315 320 Gly Leu Asn Pro Ser Ser Leu Arg Leu Ile Tyr Phe Ser Gly Asp Tyr 325 330 335 Leu Leu Lys Gly Phe Arg Ile Gly Val Val Ser Gly Met Val Ala Leu 340 345 350 Thr Glu Ala Val Ala Ile Gly Arg Thr Phe Ala Ala Met Lys Asp Tyr 355 360 365 Gln Ile Asp Gly Asn Lys Glu Met Val Ala Leu Gly Ala Met Asn Val 370 375 380 Ile Gly Ser Met Thr Ser Cys Tyr Val Ser Thr Gly Ser Phe Ser Arg 385 390 395 400 Ser Ala Val Asn Phe Met Ala Gly Cys Gln Thr Ala Val Ser Asn Ile 405 410 415 Ile Met Ser Ile Val Val Leu Leu Thr Leu Leu Phe Leu Thr Pro Leu 420 425 430 Phe Lys Tyr Thr Pro Asn Ala Ile Leu Ala Ala Ile Ile Ile Asn Ala 435 440 445 Val Ile Pro Leu Val Asp Val Asn Ala Thr Ile Leu Ile Phe Lys Ile 450 455 460 Asp Lys Leu Asp Phe Val Ala Cys Met Gly Ala Phe Phe Gly Val Ile 465 470 475 480 Phe Val Ser Val Glu Ile Gly Leu Leu Ile Ala Val Gly Ile Ser Phe 485 490 495 Ala Lys Ile Leu Leu Gln Val Thr Arg Pro Arg Thr Ala Ile Leu Gly 500 505 510 Lys Ile Pro Gly Thr Ser Val Tyr Arg Asn Ile Asn Gln Tyr Pro Glu 515 520 525 Ala Thr Arg Ile Pro Gly Val Leu Thr Ile Arg Val Asp Ser Ala Ile 530 535 540 Tyr Phe Ser Asn Ser Asn Tyr Val Arg Glu Arg Ile Gln Arg Trp Leu 545 550 555 560 Thr Asp Glu Glu Glu Met Val Glu Ala Ala Arg Leu Pro Arg Ile Gln 565 570 575 Phe Leu Ile Ile Glu Met Ser Pro Val Thr Asp Ile Asp Thr Ser Gly 580 585 590 Ile His Ala Leu Glu Asp Leu Tyr Lys Ser Leu Gln Lys Arg Asp Ile 595 600 605 Gln Leu Val Leu Ala Asn Pro Gly Pro Pro Val Ile Asn Lys Leu His 610 615 620 Val Ser His Phe Ala Asp Leu Ile Gly His Asp Lys Ile Phe Leu Thr 625 630 635 640 640 Val Ala Glu Ala Val Asp Ser Cys Ser Pro Lys Leu Ser Asp Glu Val 645 650 655

[Brief description of the drawings]

Figure 1: Sulfate transporter gene SUL1 of yeast
And SUL2 deleted yeast mutant CP154-7A is a photograph showing a growth state when the expression product of Sultr1; 3 is constantly expressed and grown in a medium using sulfate ions as a sulfur source.

FIG. 2A is a micrograph of the root of a transformed Arabidopsis thaliana, and B is a photographic image (left) and a fluorescent photographic image (right) of a cross section of the root. Ph: phloem, x: xylem, ed: endothelium, c: cortex, ep: epidermis.

Continued on the front page F term (reference) 2B030 AD07 AD08 CA14 CA19 4B024 AA08 BA80 CA04 CA07 CA11 DA01 EA01 FA02 FA15 GA11 4B065 AA89X AB01 AC20 BA01 CA24 CA53

Claims (6)

[Claims] 1. Arabidopsis thalian
a) a gene which is present on the chromosome 1 of a) and encodes a sulfate ion transporter Sultr1; 3 having an amino acid sequence of SEQ ID NO: 2; 2. The gene according to claim 1, wherein the cDNA has the nucleotide sequence of SEQ ID NO: 1. 3. The genomic DNA, mRNA and c of the gene of claim 1.
A polynucleotide purified from DNA or a sequence complementary thereto. 4. An oligonucleotide having a base sequence of 10 base pairs or more, which hybridizes to the gene of claim 1 or the polynucleotide of claim 3. 5. A recombinant vector carrying the polynucleotide of claim 3. 6. A plant into which the gene of claim 1 has been introduced, a progeny thereof, or a tissue thereof.