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Phosphorus transportprotein gene GmPT5 related to phosphorus transport of soybean nodulation and application thereof

A technology of soybean root nodule and transporter protein, applied in the field of plant biology

Inactive Publication Date: 2012-10-31
SOUTH CHINA AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Plant phosphorus transporters, especially the Pht1 family proteins, play an important role in the uptake and transport of plant phosphorus. However, there is no relevant report on whether there are two pathways for phosphorus transporters to mediate the acquisition of phosphorus in leguminous crops.

Method used

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  • Phosphorus transportprotein gene GmPT5 related to phosphorus transport of soybean nodulation and application thereof
  • Phosphorus transportprotein gene GmPT5 related to phosphorus transport of soybean nodulation and application thereof
  • Phosphorus transportprotein gene GmPT5 related to phosphorus transport of soybean nodulation and application thereof

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0048] GmPT5 gene cloning

[0049] 1. Gene ( GmPT5 ) clone

[0050] In the published soybean genome database, a total of 14 members of the soybean Pht1 phosphorus transporter family were predicted by homologous comparison. GmPT5 The gene is a low-phosphorus-induced, high-affinity phosphorus transporter gene specifically expressed in root nodules. According to the TRIzol one-step method, the total RNA of soybean root nodules of HN98 high-efficiency phosphorus genotype was extracted, and the whole genome cDNA was obtained by reverse transcription as a template, and upstream primers were used:

[0051] 5'-ATGGGGAAGGAGCAAGTTCAGG-3' (SEQ ID NO:3)

[0052] and downstream primers:

[0053] 5'- TTACACCTTGGTCTCCTCTTCTTG-3' (SEQ ID NO: 4)

[0054] For primers, perform PCR amplification. The PCR reaction system is 50 μL, including 1 μL of 10 μM forward and reverse primers, 5 μL of 10×PCR buffer, 8 μL of 2.5 mM dNTP, 0.3 μL of Taq enzyme, and 1 μL of cDNA template. Then use sterile...

Embodiment 2

[0086] Study on Transgenic Compound Plants

[0087] 1. Obtaining of transgenic compound plants

[0088] The constructed expression vector plasmid was transformed into Agrobacterium rhizogenes K599, and the transgenic composite plants (transgenic hairy root in the root and non-transgenic in the shoot) were obtained by Agrobacterium-mediated soybean hypocotyl injection. This strain was used for type identification. Empty vector control: According to the above method, the expression vector pYLRNAi was transformed into soybean in the same way to obtain a pYLRNAi-transformed empty vector control line (CK).

[0089] 2. Detection of transgenic compound plants

[0090] 15 days after inoculation, the root samples were first taken to extract RNA, and the transgenic hairy roots were screened for hygromycin, and the hygromycin resistance gene ( Hyg ) to detect the authenticity of hair roots; 40 days after inoculation, when the root nodules were in the mature stage, two root nodules we...

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Abstract

The invention discloses a phosphorus transportprotein gene GmPT5 related to phosphorus transport of soybean nodulation and application thereof. The phosphorus transportprotein gene GmPT5 related to the phosphorus transport of the soybean nodulation has a nucleotide sequence as shown in SEQ ID NO:1, and coded protein nucleotide sequence is shown as SEQ ID NO:2. The gene GmPT5 regulates and controls the phosphorus transport from a root part to the nodulation, furthermore, the overexpression of the gene is favorable for increasing the size of the soybean nodulation and the plant biomass as well as the nitrogen and phosphorus contents.

Description

technical field [0001] The invention relates to the field of plant biotechnology, in particular to a phosphorus transporter gene related to phosphorus transport in soybean nodules GmPT5 and its application. Background technique [0002] Soybean, an important oil crop and protein source, can form a mutually beneficial symbiotic system with rhizobia in the soil to fix nitrogen (N) in the air and provide a continuous nitrogen source for the host plant (Unkovich and Pate, 2000). With the requirement of agricultural sustainable development and environmental protection, using rhizobia to inoculate soybean to reduce its dependence on nitrogen fertilizer is of great significance to the scientific cultivation of soybean and the protection of the environment. Phosphorus (P) is an essential nutrient element for plant growth, and it is also an element with high demand in the growth of legumes and the process of biological nitrogen fixation. Nodule nitrogen fixation is a highly energy-...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/29C12N15/63C12N1/15C12N1/19C12N1/21C12N15/11C07K14/415A01H5/00
Inventor 廖红赵静田江秦璐
Owner SOUTH CHINA AGRI UNIV
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