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Low-phosphorus-resisting key gene GmPHR25 in plant phosphorus signal network and applications of low-phosphorus-resisting key gene GmPHR25

A signaling network, low-phosphorus-resistant technology, applied in the field of plant biology, to achieve the effect of important market prospects

Active Publication Date: 2017-12-05
SOUTH CHINA AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

in soybeans GmPHRs There are 35 members of the family, but no information about soybean GPHR Research Reports Involved in Phosphorus Signaling Networks

Method used

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  • Low-phosphorus-resisting key gene GmPHR25 in plant phosphorus signal network and applications of low-phosphorus-resisting key gene GmPHR25
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  • Low-phosphorus-resisting key gene GmPHR25 in plant phosphorus signal network and applications of low-phosphorus-resisting key gene GmPHR25

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0049] Example 1 GmPHR25 Gene cloning and vector construction

[0050] 1. Excess (OE -GmPHR25- pYL) expression vector construction

[0051] (1) design GmPHR25 Gene-specific primer OE- GmPHR25 -pYL-F and OE- GmPHR25 -pYL -R:

[0052] Primer OE-GmPHR25-pYL-F (SEQ ID NO.3):

[0053] 5'-GAGCTCATGTATCATTCAAAGAATGTTCCTAG-3'

[0054] Primer OE- GmPHR25 -pYL-R (SEQ ID NO.4):

[0055] 5'-GACGTCTCACAGATTACCGCCACC-3'.

[0056] (2) PCR amplification: the root cDNA of soybean genotype YC03-3 treated with phosphorus deficiency was used as a template, and the gene-specific primer OE- GmPHR25 -pYL-F (SEQ ID NO. 3) and OE- GmPHR25 -pYL-R (SEQ ID NO.4) amplifies GmPHR25Coding region fragment. The PCR reaction system is 50 microliters, including 5 microliters of 10×LaTaq Buffer, 4 microliters of 2.5 mmol / L dNTP, 3 microliters of cDNA template, and 1 microliter of 10 micromol / liter forward and reverse primers , 0.5 microliters of La Taq enzyme, and finally make up to 50 microliters...

Embodiment 2

[0067] Example 2 GmPHR25 Analysis of gene expression patterns and protein subcellular localization

[0068] 1, GmPHR25 Gene expression pattern analysis

[0069] (1) Experimental method

[0070] Select YC03-3 seeds with undamaged seed coat and uniform size, and use 3% hydrogen peroxide (H 2 o 2 ) surface disinfection for one minute, then rinsed twice with deionized water, soaked in 1 / 4 Hoagland nutrient solution for half an hour, and carried out paper culture to accelerate germination. After 5 days, transfer to soybean complete nutrient solution (+P) and phosphorus-deficient nutrient solution (-P). After 14 days of treatment, fully expanded new leaves and root samples were collected, and total RNA was extracted for quantitative PCR analysis.

[0071] Fluorescent quantitative PCR was carried out according to the instructions of the SYBR Green (Promega, USA) quantitative kit, and was run with the Rotor-Gene 3000 qRT-PCR system (Corbett Research, Australia). The cDNA sample...

Embodiment 3

[0085] Example 3 Research on transgenic materials

[0086] 1. Obtaining genetically modified materials

[0087] (1) Obtaining soybean hairy roots in vitro

[0088] Overexpress the constructed OE -GmPHR25- The pYL vector and the empty vector plasmid were transformed into Agrobacterium rhizogenes K599 by freeze-thaw method. Pick positive clones and inoculate them in YEP medium containing corresponding resistance, and culture them at 28°C and 200 rpm for 16 hours. Cut off the radicle of soybeans germinated on MS medium for 4 days with a scalpel dipped in bacteria solution, leaving about 3 mm of hypocotyl and divide it into two from the middle with a scalpel, and then dip the cotyledon and hypocotyl with bacteria solution Gently slit the wound with a scalpel. The cotyledons were placed on moist sterilized filter paper with the incision facing up, and cultured in the light at 25°C for 3 days. The cotyledons were then transferred to hairy root induction medium for culture. Af...

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Abstract

The invention discloses cloning and applications of a key regulation gene GmPHR25 in a plant phosphorus signal network. The nucleotide sequence of the gene is as shown in SEQ ID NO.1, and the amino acid sequence of coded protein is as shown in SEQ ID NO.2. The research shows that overexpression of GmPHR25 can increase the concentrations of soluble phosphorus and total phosphorus of soybean isolated hairy roots and compound plants under treatment of high and low phosphorus, the biomass is improved under the condition of low phosphorus, the biomass of the isolated hairy roots and the compound plants is reduced under the condition of high phosphorus, therefore, GmPHR25 can regulate the growth of transgenic soybeans and the dynamic balance of phosphorus in the body of gene soybeans, and plays an important role in adapting to low-phosphorus stress of plants; GmPHR25 can regulate the adaptation of plants to low-phosphorus stress in soil through a transgenic technology, and also can be used for genetic improvement of leguminous crops for adapting to acid soil, thus having a very important market prospect.

Description

technical field [0001] The invention belongs to the field of plant biotechnology. More specifically, it relates to a key regulatory gene for tolerance to low phosphorus in a plant phosphorus signaling network GmPHR25 and its applications. Background technique [0002] Phosphorus is an important nutrient element necessary for plant growth and development, and participates in multiple physiological, biochemical and metabolic processes of plant cells (Vance et al ., 2003; Richardson et al ., 2009). However, phosphorus in soil mainly exists in the form of insoluble inorganic phosphorus and organic phosphorus, which are difficult to be directly absorbed and utilized by plants. Low soil phosphorus availability has become an important factor limiting crop growth and production (Beardsley, 2011; Veneklaas et al .,2012). In order to maintain crop yields, a large amount of phosphate fertilizers are used in agricultural production, which has caused serious environmental pollut...

Claims

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Application Information

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IPC IPC(8): C12N15/29C07K14/415C12N15/82A01H5/00
CPCC07K14/415C12N15/8271
Inventor 田江薛迎斌朱胜男
Owner SOUTH CHINA AGRI UNIV
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