CN103266130B - Soybean water channel protein gene GmPIP1; The application of 2 - Google Patents

Abstract

The invention belongs to the field of plant genetic engineering. Specifically, the present invention relates to a soybean PIP (Plasma? Intrinsic? Protein) gene cloned by PCR, and obtained overexpression and interference materials through transgenic technology; also relates to the use of the gene to regulate soybeans to be subjected to abiotic stress tolerance so as to enhance soybean Growth and development under adverse conditions increase crop yields. The invention discloses the application of a soybean aquaporin gene PIP1;2: it is used to construct a transgenic soybean, and the transgenic soybean has abiotic stress tolerance; the soybean aquaporin gene PIP1;2 has an EQ? ID? NO: the nucleotide sequence shown in 1. The present invention specifically uses SEQ? ID? The gene of the nucleotide sequence shown in NO: 1 is transformed into soybean cotyledon nodes, and then the transformed soybean cells are cultivated into transgenic plants. <!--1-->

Description

Application of Soybean Aquaporin Gene GmPIP1;2

technical field

The invention belongs to the field of plant genetic engineering. Specifically, the present invention relates to a soybean PIP (Plasma Intrinsic Protein) gene cloned by PCR, and obtained overexpression and interference materials through transgenic technology; Growth and development under conditions increase crop yields.

Background technique

In the soil-plant-atmosphere continuous system, the water absorbed by the root system can be continuously transported to the aboveground part through the water potential difference. There are three pathways for water to move rapidly in plant root xylem: the apoplast pathway, the symplast pathway and the transcellular pathway. The symplast and transcellular pathways are difficult to separate experimentally, so they are collectively referred to as the cell-to-cell pathway. Plants change the main flow path of water in these two pathways through the difference of relative fluid conductivity and osmotic pressure gradient. The apoplast pathway can be altered by changes in the structure of the plant root, including Caspian's bands and suberin. While the rate of water transfer in the cell-to-cell pathway can be largely determined by the activity of aquaporins (AQPs) on the cell membrane, these changes are rapid and reversible. Aquaporin accelerates the rate of intercellular water migration by reducing the resistance of water transmembrane movement, and the embedding of aquaporin greatly improves the permeability of biomembrane to water. Aquaporins are members of the major intrinsic protein (MIP) family on cell membranes. They are highly hydrophobic proteins comprising six transmembrane domains with molecular weights ranging from 26-34 kD. These proteins are divided into four major subfamilies: plasma membrane intrinsic proteins (PIPs), tonoplast intrinsic proteins (TIPs), NOD26-like intrinsic proteins (NIPs), small basic intrinsic proteins (SIPs) and recently newly reported GIP, HIP and subfamilies such as XIP.

Global water scarcity is currently worsening, and large swaths of land on all continents are experiencing severe damage to ecosystems, especially soil quality and environmental quality. Improving efficient agricultural cultivation on limited land resources is high on the agenda. Soybean is one of the most important oil crops in the world, and the global demand continues to grow. It is very important to enhance soybean growth and increase soybean yield. Plants can activate the expression and activity of aquaporins under abiotic stress stress conditions. Expression of heterologous aquaporins in Arabidopsis or tobacco can enhance the vitality or stress tolerance of transgenic plants (Aharon et al. 2003, Gao 2010, Hu et al. 2012, Katsuhara et al. 2003 ), proposed Overexpression of ginseng vacuolar aquaporin PgTIP1 in A. thaliana promoted the growth and development of transgenic A. thaliana and increased its seed size (Lin et al. 2007).

Soybean is an important economic crop in the world, but the research on its aquaporin is still blank. Since the sequencing of the whole soybean genome was completed and released in 2010, 21 genes suspected of soybean aquaporin were found in the entire published genome, and the research on the function and mechanism of these genes is still blank.

The above references are as follows:

1. Aharon R, Shahak Y, Wininger S, Bendov R, Kapulnik Y, and Galili G (2003) Overexpression of a plasmamembrane aquaporin in transgenic tobacco improves plant vigor under favorable growth conditions but not under drought or salt stress. Plant Cell. 15 :439-447 (Aharon R, Shahak Y, Wininger S, Bendov R, KapulnikY, and Galili G (2003) Overexpression of a plasma membrane aquaporin in transgenic tobacco increases plant vigor under favorable growth conditions, but not Its drought resistance or salt resistance. Plant Cell. 15:439-447);

2. Gao Z, He X, Zhao B, Zhou C, Liang Y, Ge R, Shen Y, and Huang Z (2010). Overexpressing a putative aquaporin gene from wheat, TaNIP, enhances salt tolerance in transgenic Arabidopsis. Plant Cell Physiol51, 767-775 (Gao Z, He X, Zhao B, Zhou C, Liang Y, Ge R, Shen Y, and Huang Z (2010). Expression of the suspected wheat water channel gene TaNIP in Arabidopsis, increased transgenic Arabidopsis Salt resistance. Plant CellPhysiol 51, 767-775.);

3. Hu W, Yuan Q, Wang Y, Cai R, Deng X, Wang J, Zhou S, Chen M, Chen L, Huang C, Ma Z, Yang G, and He G (2012). Overexpression of a Wheat Aquaporin Gene , TaAQP8, Enhances Salt Stress Tolerance in Transgenic Tobacco. Plant Cell Physiol 53, 2127-2141 (Hu W, Yuan Q, Wang Y, Cai R, Deng X, Wang J, Zhou S, Chen M, Chen L, Huang C, Ma Z, Yang G, and HeG (2012) Expression of the wheat water channel gene TaAQP8 in tobacco increases the salt tolerance of transgenic tobacco. Plant Cell Physiol 53, 2127-2141);

4. Katsuhara M, Koshio K, Shibasaka M, Hayashi Y, Hayakawa T, and Kasamo K. (2003) Over-expression of abarley aquaporin increased the shoot root ratio and raised salt sensitivity in transgenic rice plants. Plant Cell Physiol 44: 1378- 1383 (Katsuhara M, KoshioK, Shibasaka M, Hayashi Y, Hayakawa T, and Kasamo K. (2003) Overexpression of water channel genes in barley increases salt tolerance and shoot-to-root ratio in transgenic rice. Plant Cell Physiol 44 : 1378-1383 .);

5. Lin WL, Peng YH, Li GW, Arora R, Tang ZC, Su WA, and Cai W. (2007) Isolation and functional characterization of PgTIP1, a hormone-autotrophic cells-specific tonoplast aquaporin in ginseng. J Exp Bot 58 :947-956 (Lin WL, Peng YH, Li GW, Arora R, Tang ZC, Su WA, and Cai W. (2007) Cloning and functional analysis of vacuolar aquaporin PgTIP1 in hormonoautotrophic cells from Panax ginseng. J Exp Bot 58: 947-956.).

Contents of the invention

The technical problem to be solved by the present invention is to provide a protein and its gene capable of making soybean have abiotic stress tolerance and biomass increase, as well as the transgenic plant obtained therefrom, and a method for transforming soybean by using the gene.

In order to solve the above technical problems, the present invention provides a use of soybean aquaporin gene PIP1;2: for the construction of transgenic soybeans, which have abiotic stress tolerance;

The soybean aquaporin gene PIP1;2 has the nucleotide sequence shown in EQ ID NO:1.

As an improvement of the use of the soybean aquaporin gene PIP1;2 of the present invention: the transgenic soybean has salt tolerance (salt resistance).

As an improvement of the use of the soybean aquaporin gene PIP1;2 of the present invention: the transgenic soybean has the characteristic of increasing the biomass.

As an improvement on the application of the soybean aquaporin gene PIP1;2 of the present invention: the soybean cotyledon node is transformed with the gene having the nucleotide sequence shown in SEQID NO: 1, and then the transformed soybean cells are cultivated into transgenic plants.

The new gene PIP1;2 cloned from the soybean wild-type variety Williams 82 provided by the present invention---the plasma membrane intrinsic protein gene PIP1;2 in the soybean aquaporin family, has as shown in SEQ ID NO:1 DNA sequence (nucleotide sequence). The sequence number of the PIP1;2 gene is Gm08g01860.

The gene PIP1;2 encodes a soybean aquaporin located in the plasma membrane, has the amino acid sequence shown in SEQ ID NO: 2, and belongs to the MIP gene family.

To be more specific:

Another object of the present invention is to provide a kind of method that carries out efficient soybean transformation with PIP gene, specifically, the present invention provides the vector that has the gene of sequence shown in Seq ID No.1 and Fig. 5 or gene partial fragment , wherein, as shown in Figure 1 PIP1; 2-oe, the vector can express the polypeptide encoded by the above nucleotide sequence.

The invention also provides a method for transforming plants with plant expression vectors to affect the ability of soybean aquaporin PIP content.

Realize the concrete technical steps of the present invention as follows:

1. Cloning of soybean PIPs genes

The soybean PIP1; 2 gene was cloned by PCR technology, and the two genes were constructed into soybean transformation expression vectors by molecular biology techniques such as restriction endonuclease ligation. see picture 1.

2. Soybean Genetically Modified

Transgenic plants with overexpression of PIP1;2 gene were obtained by using Agrobacterium-mediated rapid preparation of soybean transgenic receptors.

3. Regulate the expression of PIP gene in soybean:

Through the expression of PIP1;2 gene in soybean by transgenic technology, the overexpressed and interfered transgenic soybean was obtained, and the transgenic plants were detected by glufosinate smear (Figure 2), semi-quantitative RT-PCR (Figure 3) and other methods.

4. Preliminary identification of PIP gene function

The transgenic overexpression material and wild-type soybean were cultivated in soybean nutrient solution, and the seedlings grown to 10 days old were treated with 100mM Nacl for 7 days to observe the phenotype. Inhibited, leaves yellowed (Fig. 4).

At present, there is a crisis of decreasing arable land area and shortage of water and non-renewable fertilizer resources in our country. There is an urgent need to breed soybean varieties with strong tolerance, high yield and high efficiency. Aquaporins are the basis for determining soybean yield and water nutrient absorption efficiency. The development of genetic engineering technology makes it possible to adjust soybeans to adapt to different adversity conditions by regulating PIP genes. The invention obtains soybean aquaporin gene PIP through cloning technology, obtains overexpression and interference materials through transgene, and preliminarily identifies the function of the gene. Therefore, the invention can make soybeans have good adversity tolerance, thereby promoting the growth and development of soybeans, and finally improving the yield of soybeans.

The present invention clarifies the functions of soybean aquaporin GmPIP1;2 in salt resistance and yield improvement. The achievement of the invention can be applied to regulating the expression of the soybean aquaporin gene through biotechnology, thereby cultivating new soybean varieties with significantly improved yield or resistance to environmental stress. The invention has good application prospect.

Description of drawings

The specific implementation manners of the present invention will be described in further detail below in conjunction with the accompanying drawings.

Fig. 1 is soybean transformation expression vector PIP1; 2-oe carrier map;

The vector uses the bar gene as a transgenic screening marker, and the PIP1;2-oe vector uses the 35S promoter followed by the full-length GmPIP1;2 cDNA, which can be used to obtain transgenic plants overexpressing GmPIP1;2 after transgenesis.

Figure 2 shows the test results of the glufosinate leaf smear method on transgenic soybeans; half of the leaves marked in black indicate that no herbicide was applied; the leaves on the other side of the main vein were coated with 135mg/L Basta herbicide. Non-GMO soybeans on the left; GMO soybeans on the right.

Figure 3 is the result of semi-quantitative RT-PCR identification of PIP1;2 gene expression in soybean transgenic plants; the upper picture in the figure is the PCR product of PIP1;2 gene, and the lower picture is the PCR product of the soybean housekeeping gene-actin-encoding gene Actin ( used as an internal reference). WT is non-transgenic soybean, 1-11 are 11 independent transgenic lines with overexpression of PIP1;2 gene.

Fig. 4 is the salt tolerance identification result of PIP1; 2 overexpression soybean; Soybean seedlings 10 days after germination are transferred to the culture solution containing 100mM NaCl and grown for 7 days as salt treatment; Transgenic soybean (control) and PIP1;2 gene overexpression transgenic soybean plant (PIP1;2-Oe); the right picture shows non-transgenic soybean (control) and PIP1;2 gene overexpression transgenic soybean plant (PIP1;2) after 7 days of salt treatment -Oe) plant height and aboveground biomass.

Fig. 5 is the DNA sequence of PIP1; 2 gene;

Fig. 6 is the amino acid sequence encoded by PIP1;2 gene.

Detailed ways

Embodiment 1, soybean PIP1; Cloning of 2 gene

(1) Extraction of total RNA: Take 50-100 mg of leaves of Williams 82 soybean seedlings at the true leaf stage, and use Trizol method to extract total RNA. The specific method is to grind soybean leaf samples into powder in liquid nitrogen, then add 1 ml of Trizol (Takala Company), thaw the ground samples at room temperature, and transfer them to 200 microliters of chloroform solution after the samples are completely "damp". Shake vigorously to dissolve it completely in the lysate; shake it at room temperature for 5 minutes and then centrifuge at 12000 g for 10 minutes at 4 °C. Aspirate the upper colorless aqueous phase, add an equal volume of isopropanol, slowly invert and mix well, leave at room temperature for 10 minutes, and then centrifuge at 12000 g for 10 minutes. Add 1 ml of 75% ethanol to the RNA pellet, wash thoroughly, centrifuge, dry, and add 30 µl of DEPC water to dissolve the precipitate.

(2) Preparation of cDNA: The first strand of cDNA was synthesized using a reverse transcription kit. Take a 1.5 ml RNase-free centrifuge tube, add 4 μg of RNA, 2 μl of 10 μg/ml Oligo-dT (Promega) and a certain volume of DEPC water to a final volume of 15 μl. Mix the reaction solution, denature it in a water bath at 70°C for 5 minutes, then immediately cool it on ice for 10 minutes; add 8 μl of FirstStrand buffer (Promega), 3 μl of 10 mM dNTP, 0.5 μl of RNase inhibitor (40 unit), and 1 microliter of M-MLV reverse transcriptase, the final volume of the reaction was 40 microliters, the reaction was terminated at 75℃ for 10 minutes after reacting at 42℃ for 1 hour.

(3) Reverse transcription PCR amplification of GmPIP1;2 full-length cDNA gene: the PCR primer sequences used to amplify GmPIP1;2 cDNA are: upstream primer: GGACTTTGAACTACACTACA; downstream primer: CTTTCTATTGAAGCCGCCTT. The PCR reaction system is: 2 μl of cDNA, 0.8 μl of 2.5mM dNTP, 2 μl of 10xPCR buffer, 0.2 μl of 10 μM upstream primer and 0.2 μl of downstream primer, 0.3 μl of Taq enzyme, 14.5 μl of water, the total volume is 20 microliter. The PCR reaction program was: 94°C, 5min; 94°C, 30s; 56°C, 1min; 72°C, 30s; after 32 cycles, 72°C, 10min. After the PCR reaction, a 1% agarose gel was made, and the PCR product was spotted and electrophoresed for 20 minutes. The DNA product was recovered with a gel recovery kit, and sequenced to confirm that the cDNA sequence was completely consistent with EQ ID NO:1.

Embodiment 2, GmPIP1; The construction of transgenic PIP1 of 2 genes; 2-Oe carrier

The vector is used for the overexpression of GmPIP1;2 gene in soybean. The full-length cDNA of GmPIP1;2 was connected to the binary vector pBAR, which can be used for Agrobacterium-mediated transgene, with the restriction endonuclease site SmalI. The pBAR vector carries a selection marker gene bar in the T-DNA region, which encodes glufosinate acetyl CoA transferase (PAT), which can catalyze the free aminoacetylation of glufosinate, thereby inactivating the herbicide glufosinate. GmPIP1;2 is driven by the 35S promoter. The final vector PIP1;2-Oe map is shown in Figure 1.

Embodiment 3, GmPIP1; The cultivation of 2 gene overexpression transgenic soybean

The original recipient material of soybean transgene was Williams 82. The transgenic method is as follows:

(1). Seed disinfection: The surface disinfection of soybean seeds is sterilized by chlorine gas dry method. First, select clean seeds that are mature and plump, without disease spots, and without hardness, and arrange them in a single layer in a 90*15mm petri dish; place the petri dish Open the cover and put it in the desiccator. Place a 500ml glass beaker in the desiccator. Use a 100ml measuring cylinder to measure 75ml of commercial bleach and add it to the beaker. Measure 3ml of 12N HCl with a 10ml measuring cylinder and add it slowly along the wall of the beaker; cover it The lid of the desiccator, to ensure that the container is sealed, let it stand overnight for 10-16 hours; after the sterilization is completed, transfer the petri dish to a sterile ultra-clean table, open the lid of the petri dish, and blow it with strong wind for 25-40 minutes to remove Chlorine gas remains. Surface-sterilized dry seeds can be stored airtight at room temperature for about 2 weeks.

(2). Agrobacterium preparation and infection: extract the binary vector PIP1;2-Oe plasmid DNA, transfer the binary vector into Agrobacterium strain EHA101 by electroporation, and store in 50% glycerol. Two days before the transgenesis, pipette 50 μl of Agrobacterium glycerol containing the carrier into 5 ml of YEP liquid medium (10 g/L peptone, 5 g/L yeast extract, 5 g/L sodium chloride, pH7. 0), 28°C, 250rpm, shaking culture for 24-36 hours; pipette 0.2-1ml of saturated bacterial liquid into 250ml YEP liquid medium added with antibiotics (1/2000) to expand the culture until OD650nm=0.8-1.0; Divide the bacterial solution into several 50ml sterile centrifuge tubes, centrifuge (4000rpm, 10min, 25°C), collect the colonies, gently pipette with 25-50ml liquid co-culture medium (LCCM), resuspend the pellet and set aside. LCCM medium containing 1/10 B5 macro, trace and vitamin (Gamborg et al., 1968), 3% sucrose, organic buffer 2 (N-morpholine) ethanolsulfonic acid (MES) 3.9 g/L, pH 5.4 , sterilized at 120°C for 20 minutes, added gibberellin (GA3) 0.25 mg/L, 6-benzyl adenine (BAP) 1.67 mg/L, cysteine (Cys) 400 mg/L, Dithiothreitol (DTT) 154.2 mg/L, acetosyringone (As) 200 μmol/L.

(3). Infection: Place the imbibed soybean seeds on sterile absorbent paper, cut the seeds longitudinally with a scalpel along the hilum, separate the cotyledons and hypocotyls evenly, and remove the seed coat for later use. Pour the resuspension of Agrobacterium into a clean sterile petri dish, put about 50 explants, and infect at room temperature for 20-30 minutes. During this period, the bacterial solution is often stirred to make the explants fully contact with the fresh bacterial solution.

(4). Co-cultivation: After the infection, the explants were taken out, blotted dry with sterile absorbent paper, and then placed on the co-culture medium (CM) with sterile filter paper, 7 to 10 explants per dish body, with the adaxial side facing up, placed horizontally. The formulation of CM medium was the same as that of LCCM, plus 5 g/L agar (DifcoAgar, Noble Company). Stack the petri dishes, seal them with plastic wrap and place them in a Percival incubator at 23°C in the dark for 3 to 5 days.

(5). Cluster bud induction: After 3-5 days of co-cultivation, cut off the elongated hypocotyl and leave about 0.5 cm, and insert it on the bud induction (SI) medium with screening agent at an oblique angle of 30-45° , SI medium containing B5 macronutrients, micronutrients and vitamins, 30g/L sucrose, 0.59g/L MES, and 8g/L agar (Sigma, USA), after sterilizing at 120°C for 20min, add 1.67 mg of BAP under sterile conditions /L, ticarcillin (Tic) 250 mg/L, cephalosporin (Cef) 100 mg/L. Seal with 3M air-permeable tape and transfer to the culture room (24 ^o C, 18/6 light intensity 140 μ moles/m ² /sec), culture for 4 weeks, and replace with fresh SI medium every two weeks.

(6). Cluster shoot elongation: After 4 weeks of screening for cluster shoot induction, the remaining cotyledons were excised and transferred to shoot elongation (SE) medium, which contained MS macronutrients, micronutrients and vitamins (Murashige and Skoog, 1962 ), sucrose 30 g/L, MES 0.59 g/L, agar (Sigma, USA) 8 g/L, pH 5.8, sterilized at 120°C for 20 min, GA 30.5 mg/L, L- Asparagine (L-Asp) 50 mg/L, glutamine (Glu) 50 mg/L, indole acetic acid (IAA) 0.1 mg/L, zeatin (ZR) 1 mg/L, Tic250 mg/L and Cef100 mg/L, the culture conditions were the same as the cluster bud induction process, cultured for 2-8 weeks, and replaced with fresh SE medium every 2 weeks. The culture conditions are 24 ^o C, 18/6 light intensity 140μ moles/m ² /sec).

(7). Rooting: Cut off the young shoots with a length of 3-4 cm, soak them in indole butyric acid (IBA) for 30s~1min, and then insert them into the rooting medium (RM). The RM medium contains a large amount and a small amount of MS. and vitamins, sucrose 20 g/L, MES 0.59 g/L, agar (Sigma, USA) 8 g/L, IBA 0.1 mg/L, L-Asp 50 mg/L, Glu 50 mg/L, Tic 250 mg/L L. Cef 100 mg/L. After 1-2 weeks, when the root length is about 2-3 cm, take out the rooted seedlings from the culture medium, wash the residual medium at the root, transfer to the soil and move to the greenhouse for cultivation. The culture conditions were 24 ^o C, 18/6 light intensity 140 μ moles/m ² /sec.

(8). Herbicide resistance identification of soybean transgenic materials: because the transgenic vector encodes glufosinate acetyl-CoA transferase (PAT), it can catalyze the free aminoacetylation of glufosinate, thereby making the herbicide glufosinate inactive. live. The herbicide Liberty® for identification, also known as Basta, has a stock solution concentration of 135 g/L. In the experiment, the stock solution was diluted 1000 times, and a little 135 mg/L Basta was applied on half of the soybean leaf with a cotton swab, bounded by the main vein, and marked with a marker on the other half of the leaf to indicate that no herbicide was applied In contrast, the leaves were observed after 3-5 days. If half of the leaves coated with the herbicide were significantly shriveled compared to the control, the sample was negative (Figure 2 right); if the leaves were normal, the sample was positive (Figure 2 left).

Example 4, detection of the expression level of the target gene in transgenic soybean

The total RNA of leaves of PIP1;2-Oe transgenic soybean and non-transgenic soybean (Williams 82) was extracted by Trizol method (as described in Example 1). After reverse transcription, the expression was detected by semi-quantitative RT-PCR. Soybean housekeeping gene-actin-encoding gene Actin was used as an internal reference. According to Figure 3, the following conclusions can be drawn: in the 11 PIP1;2-Oe overexpression transgenic lines, the expression of PIP1;2 gene was significantly increased compared with the non-transgenic control (Figure 3). Therefore, the transgenic soybean has obtained the expected transgenic effect.

Embodiment 5, cultivation condition and stress resistance identification of soybean material

The harvested seeds (T1 generation) of the obtained transgenic soybean plants were dried and sterilized by chlorine gas, then sowed in sterilized humus soil, germinated and cultivated under light conditions at 30°C, and the seedlings that had grown for 5 days were carefully removed from the soil and washed Soil on the roots, transplanted to sponge balls on PVC boards and then transferred to soybean nutrient solution for cultivation.

Soybean cultivation conditions are as follows: Light and temperature controllable soybean cultivation room. Temperature control: 30°C during the day and 20°C at night. The photoperiod is 12 hours, and the light intensity: 250-300 μ moles/m ² /sec). Soybean umol.m-2.s-1, soybean culture solution pH5pH5.8-6.0, adjusted once a day, soybean culture solution replaced every 3 days.

The formula of soybean culture liquid mother liquid (i.e. stock liquid) is as follows:

Remarks: Each stock solution is prepared with water.

When in use, add 1ml of No. 1 - No. 4 stock solution and 5ml of No. 5 stock solution to each 1L of culture medium, and dilute with water.

In order to fully prove the salt-resistance performance of the transgenic soybean obtained in the present invention, the salt-resistance capability of the transgenic soybean was identified in experiments. The experiment was carried out in hydroponic conditions. 10-day-old wild-type soybean Williams82 and transgenic seedlings overexpressing PIP1;2 were treated with 100 mM NaCl for 7 days, and the treatment without NaCl was used as a control. It can be seen from Figure 4 that under salt stress, compared with the non-transgenic control soybeans, the length and fresh weight of transgenic soybeans above the ground were significantly increased, and the salt tolerance was significantly improved.

After the salt treatment, the sodium and potassium ion contents in the soybean plant leaves, stems and roots were measured respectively. It can be seen from Figure 5 that after the salt treatment, compared with the non-transgenic soybean, the sodium ion in the root of the PIP1;2 overexpression transgenic soybean has little difference, Potassium ions increased; however, it can be seen that the content of sodium ions in transgenic soybean leaves was significantly reduced, and there was no difference in potassium ions; the concentration of sodium and potassium ions in stems was not significantly different. The reduction of sodium ion in leaves of PIP1;2 overexpressed transgenic soybean is the mechanism of PIP1;2 gene salt tolerance.

Finally, it should be noted that the above examples are only some specific embodiments of the present invention. Obviously, the present invention is not limited to the above embodiments, and many variations are possible. All deformations that can be directly derived or associated by those skilled in the art from the content disclosed in the present invention should be considered as the protection scope of the present invention.

Claims (2)

1. soybean water channel protein gene PIP1; The purposes of 2, is characterized in that: for building genetically engineered soybean, and described genetically engineered soybean has abiotic stress tolerance;

Described soybean water channel protein gene PIP1; 2 is the nucleotide sequence shown in SEQIDNO:1;

Described genetically engineered soybean has salt tolerance;

Described genetically engineered soybean has the characteristic that biomass is increased.

2. soybean water channel protein gene PIP1 according to claim 1; The purposes of 2, is characterized in that: with the gene transformation soybean cotyledon node for the nucleotide sequence shown in SEQIDNO:1, then the soya cells after transforming is cultivated into transfer-gen plant.