Soybean root specific efficient phosphate transporter

A transport protein and soybean root technology, applied in the field of genetic engineering, can solve problems such as increased phosphorus content and environmental pollution, and achieve the effects of improving utilization rate, increasing yield, and improving stress resistance

Inactive Publication Date: 2012-10-03
INST OF CROP SCI CHINESE ACAD OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

As a result, the phosphorus content in the soil and water bodie

Method used

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  • Soybean root specific efficient phosphate transporter
  • Soybean root specific efficient phosphate transporter
  • Soybean root specific efficient phosphate transporter

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0025] Cloning of embodiment 1 GmPHT06 gene

[0026] Soybean cultivar Kennong 18 (KNl8) was used as the experimental material, under low phosphorus stress ( 100 μM) in liquid medium until the true leaves opened, and the roots, stems and leaves were taken, and the total mRNA was extracted using the Trizol kit (Invitrogen Company), and the reverse transcription kit (Takara Company) was used to reverse the total mRNA. Transcribed to obtain cDNA.

[0027] Liquid medium composition:

[0028]

[0029]

[0030] Using the soybean variety Kennong 18 (KN18) as the test material, RT-PCR (the program is: 95°C pre-denaturation for 3 minutes; 94°C for 30s, 55°C for 30s, 72°C for 2min, 26 cycles; 72°C for 5min. Reaction system: 0.3 μL LA Taq DNA polymerase, 1 μL primer F, 1 μL primer R, 5 μL 10X buffer, 4 μL dNTP mix, 2 μL cDNA, 36.7 μL H 2 O; the total system is 50 μL. Among them, primer F: 5-ATGGCTGGAGAATTGGGAGTTT-3; primer R: 5'-GGAACAGGAACTGTCCTAGCAG-3'. GmPHT06 was amplified...

Embodiment 2

[0032] Example 2 Expression of GmPHT06 gene in yeast

[0033]GmPHT06 was recombined into the pYES-DEST52 vector (purchased from Invitrogen) through the Gateway cloning system, and the recombinant plasmid was transformed into a yeast mutant PAM2 (Δpho89::TRP1Δpho84) lacking two high-affinity phosphorus transport genes (PHO89 and PHO84). ::HLS3 ade2 leu2 his3 trp1 ura3. Martinez, P., Zvyagilskaya, R., Allard, P and Persson, B. 1998. Physiological regulation of the derepressible phosphate transporter in Saccharomyces cerevisiae. J. Bacteriol. 180, 2253-2256. )middle. Under the action of the GAL (galactose) promoter, GmPHT06 can complement the functions of PHO84 and PHO89, that is, PAM2 can grow normally under low phosphorus (<50 μM) conditions.

Embodiment 3

[0034] Example 3 Expression of GmPHT06 Gene in Soybean and Its Application in Improving Phosphorus Utilization of Soybean

[0035] Soybean KN18 with full and consistent grains was selected as the test material. Sow it in new vermiculite to moisturize and accelerate germination, and analyze the organs at each growth stage; the phosphorus stress experiment is to germinate soybeans in the soil for 2-3 days, and then move them into In 500μM liquid medium, cultivate until the true leaves open. move to Cultured in medium with a concentration gradient of 5 μM. After cultivating for 7 days, samples were taken, and the operation was the same as in Example 1.

[0036] The regulation of GmPHT06 by phosphorus stress was detected by qRT-PCR. Primer F: 5'-TTGGTTGTGACGTGTTTACACC-3'; Primer R: 5'-ACAAAAGTAAGAACCAAAAGCAAAC-3'.

[0037] PCR program:

[0038] ABI StepOne was performed and SYBR Green I was used to detect the fluorescent signal. A 15 μl reaction system was used, and the s...

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Abstract

The invention belongs to the field of genetic engineering, and relates to a soybean root specific efficient phosphate transporter GmPHT06 whose amino acid sequence is shown by SEQ ID No.2. The soybean root specific efficient phosphate transporter also has genes encoding the protein whose nucleotide sequence is shown by SEQ ID No.1. The soybean root specific efficient phosphate transporter GmPHT06 provided by the invention can be used to improve the phosphorus utilization of various plants and resistance ability of plants under phosphorus stress, so as to increase the yield of plants.

Description

technical field [0001] The invention belongs to the field of genetic engineering, in particular to a soybean transport protein and its application. Background technique [0002] Phosphorus is one of the important nutrients for the growth and development of organisms. It is an important component of proteins, nucleic acids, lipids and various important small molecules (such as energy supplier ATP). It is an essential intermediary for various metabolisms (such as sugar degradation) substances that are regulators of growth and development (such as phosphorylation). Therefore, the application of phosphorus fertilizer plays a key role in improving agricultural yield. However, phosphorus is a non-renewable resource. Due to long-term overuse, phosphorus is gradually becoming a mineral resource that is about to disappear on the earth. In the near future, phosphorus will restrict the economic and political lifelines of various countries. Phosphorus It will also be a strategic asset...

Claims

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Application Information

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IPC IPC(8): C07K14/415C12N15/29C12N15/63C12N5/10C12N1/19C12N15/82A01H5/00
Inventor 傅永福范成明张晓玫
Owner INST OF CROP SCI CHINESE ACAD OF AGRI SCI
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