Low-phosphorus resistant gene and application thereof

A low-phosphorus and environmental-resistant technology, applied in the fields of biotechnology and botany, can solve the problems that plants cannot be directly used, water eutrophication and ecological environment pollution, and aggravate the depletion of phosphate rock resources.

Inactive Publication Date: 2012-08-01
SHANGHAI INST OF BIOLOGICAL SCI CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The total amount of phosphorus in the soil is abundant, but it is easily fixed by minerals and organic matter, most of which exist in the form that plants cannot directly use, making phosphorus deficiency one of the main factors limiting plant growth
Applying a large amount of

Method used

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  • Low-phosphorus resistant gene and application thereof
  • Low-phosphorus resistant gene and application thereof
  • Low-phosphorus resistant gene and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0165] Embodiment 1, the gene and protein sequence of AtGDPD1

[0166] Genomic DNA extraction

[0167] (1) Take an appropriate amount of plant (Arabidopsis thaliana, Col-0) tissue and place it in a 1.5ml centrifuge tube, add 50 μl of extract (0.2M Tris-HCl, pH 9.0, 0.4M LiCl, 25mM EDTA, 1% (v / v) SDS), after grinding into a slurry with a grinding rod, add 350 μl of extracting solution to rinse the grinding rod.

[0168] (2) Add 400 μl of phenol / chloroform (1:1), shake vigorously, centrifuge at 12000 rpm for 15 minutes, and take the supernatant.

[0169] (3) Add 400 μl of chloroform, shake vigorously, centrifuge at 12000 rpm for 15 minutes, and take the supernatant.

[0170] (4) Add 500 μl of isopropanol, mix well, let stand for 10 minutes, centrifuge at 12000 rpm for 10 minutes, and discard the supernatant.

[0171] (5) Wash the precipitate once with 70% (v / v) ethanol, and centrifuge at 12000 rpm for 5 min.

[0172] (6) Discard the supernatant, blow dry, and add 20-50 μl of...

Embodiment 2

[0208] Example 2, AtGDPD1 tissue expression characteristics

[0209] The extraction, reverse transcription and PCR amplification of total RNA were the same as in Example 1.

[0210] The Northern blot method is the same as described in the literature Huang, J.R., Takano, T. and Akita, S. (2000) Expression of a-expansin genes in young seedlings of rice (oryza sativa L.). Planta, 211, 467-473.

[0211] GUS detection

[0212] The method is the same as described in the literature Jefferson, R.A., Kavanagh, T.A. and Bevan, M.W. (1987) GUSfusions: β-glucuronidase as a sensitive and versatile gene fusion marker in higherplants. EMBO J.6, 3901-3907.

[0213] E. coli transformation

[0214] Escherichia coli competent cells TOP10 were purchased from Invitrogen. Transform Escherichia coli using the heat shock method, mix the plasmid with melted competent cells, place on ice for 30 minutes, heat shock in a water bath at 42°C for 45 seconds, and place on ice for 2 minutes. Add 400 μl of...

Embodiment 3

[0223] Example 3, AtGDPD1 expression is induced by low phosphorus nutrient growth environment

[0224] The inventors detected by Northern blot analysis and found that the expression of AtGDPD1 gene was strongly induced by the low-phosphorus nutrient growth environment, and as the stress time of the low-phosphorus growth environment prolongs, the expression of AtGDPD1 gene also gradually increased ( figure 2 a). This suggests that AtGDPD1 is involved in the growth of plants adapting to low phosphorus nutrient environment.

[0225] In order to further understand the molecular mechanism of the AtGDPD1 gene's response to the low-phosphorus growth environment, the inventors detected whether the promoter of the AtGDPD1 gene responded to the low-phosphorus growth environment. The result is as figure 2 As shown in b, the activity of the AtGDPD1 gene promoter was obviously induced by the low phosphorus environment. Compared with the wild type, the expression level of GUS in the who...

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Abstract

The invention relates to a low-phosphorus resistant gene and an application thereof. The invention discloses a gene which is useful to improve the capacity of plant low phosphorus resistance, that is glycerin phosphodiesterase gene. The gene can well be applied in the improvement of plant varieties to improve the resistance of the plants to the environmental stress (low phosphorus environment). The gene provides a very valuable gene resource for using transgenic technology and other molecular breeding technologies to cultivate new crop varieties which have low-phosphorus resistance and can be effectively recycled.

Description

technical field [0001] The invention belongs to the fields of biotechnology and botany; more specifically, the invention relates to a low-phosphorus-resistant environment gene and application thereof. Background technique [0002] Although applying a large amount of phosphorus fertilizers in agriculture can temporarily meet the phosphorus needs of plant growth, it will increase production costs, aggravate the depletion of phosphate rock resources, and cause eutrophication of water bodies and pollution of the ecological environment. [0003] Phosphorus is one of the essential nutrients for plant growth and development. It is a component of important substances such as phospholipids and nucleic acids in cells. It regulates life activities through protein phosphorylation and dephosphorylation, participates in various metabolic processes of sugar, protein and lipids, and plays an important biological role in the life activities of plants. learning function. The total amount of...

Claims

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Application Information

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IPC IPC(8): C12N9/16C12N15/55C12N15/82C12N15/113C12N15/63C12Q1/68A01H5/00
Inventor 黄继荣程玉祥
Owner SHANGHAI INST OF BIOLOGICAL SCI CHINESE ACAD OF SCI
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