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Application of soybean aquaporin gene GmPIP1;2

An aquaporin and soybean technology, applied in the application, genetic engineering, plant genetic improvement and other directions, can solve the problem of no increase in drought resistance or salt resistance, achieve good application prospects, improve soybean yield, and have the effect of strong tolerance

Inactive Publication Date: 2013-08-28
ZHEJIANG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] 1. Aharon R, Shahak Y, Wininger S, Bendov R, Kapulnik Y, and Galili G (2003) Overexpression of a plasma membrane 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, Kapulnik Y, and Galili G (2003) Overexpression of a plasma membrane aquaporin in transgenic tobacco increases plant vigor under favorable growth conditions, but Did not increase its drought or salt tolerance

Method used

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  • Application of soybean aquaporin gene GmPIP1;2
  • Application of soybean aquaporin gene GmPIP1;2
  • Application of soybean aquaporin gene GmPIP1;2

Examples

Experimental program
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Effect test

Embodiment 1

[0041] Embodiment 1, soybean PIP1; Cloning of 2 gene

[0042] (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". In a 1.5 ml RNase-free centrifuge tube, shake vigorously to dissolve it completely in the lysate; shake 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 di...

Embodiment 2

[0045] Embodiment 2, GmPIP1; The construction of transgenic PIP1 of 2 genes; 2-Oe carrier

[0046] 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. For the final vector PIP1;2-Oe, see figure 1 .

Embodiment 3

[0047] Embodiment 3, GmPIP1; The cultivation of 2 gene overexpression transgenic soybean

[0048] The original recipient material of soybean transgene was Williams 82. The transgenic method is as follows:

[0049] (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...

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Abstract

The invention belongs to the field of plant gene engineering. Specifically, the present invention relates to a PCR cloning soybean PIP (Plasma Intrinsic Protein) gene and an overexpression and interference material by transgenic technology; and also relates to application of the gene to regulate the tolerance of soybean to abiotic adversity stresses thus increasing soybean growth in adversity conditions and improving crop yields. The present invention discloses application of soybean aquaporin gene GmPIP1;2, which is used for constructing transgenic soybean with abiotic stress tolerance. The soybean aquaporin gene PIP1;2 has a nucleoside acid sequence shown as EQ ID NO: 1. The present invention specifically includes transforming soybean cotyledonary node by the nucleotide sequence shown as EQ ID NO: 1, and culturing the transformed soybean cells into transgenic plants.

Description

technical field [0001] 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 [0002] 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 pressur...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/82C12N15/84A01H5/00
Inventor 寿惠霞周练王创刘瑞芳韩强
Owner ZHEJIANG UNIV
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