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Plant salt tolerance related protein hbscabp10 and its coding gene and application

A technology that encodes genes and proteins, applied in plant gene improvement, application, plant peptides, etc., can solve the problems of less research on stress-resistance regulation gene cloning and functional analysis, and achieve the effect of improving salt tolerance

Active Publication Date: 2020-11-03
BEIJING ACADEMY OF AGRICULTURE & FORESTRY SCIENCES
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, previous studies were limited to morphology, anatomy, and physiology, and there were few studies on the cloning and functional analysis of stress-resistance regulatory genes.

Method used

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  • Plant salt tolerance related protein hbscabp10 and its coding gene and application
  • Plant salt tolerance related protein hbscabp10 and its coding gene and application
  • Plant salt tolerance related protein hbscabp10 and its coding gene and application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0075] Example 1. Obtaining of salt-tolerance-related proteins

[0076] 1. Acquisition of HbSOS1 protein and its coding gene

[0077] Sequence analysis of the wild barley genome reveals a Na + / H + The transporter is named as HbSOS1 protein, as shown in sequence 2 of the sequence list. The gene encoding the HbSOS1 protein is named HbSOS1 gene, and the full-length cDNA of the HbSOS1 gene is shown in sequence 1 of the sequence table, and the 71-3490th position from the 5' end of sequence 1 is an open reading frame.

[0078] 2. Acquisition of HbSCaBP10 protein and its coding gene

[0079] Sequence analysis of the wild barley genome revealed a calcium ion sensor, which was named HbSCaBP10 protein, as shown in sequence 4 of the sequence table. The gene encoding the HbSCaBP10 protein is named HbSCaBP10 gene, and the full-length cDNA of the HbSCaBP10 gene is shown in sequence 3 of the sequence table, and the 1-783 positions from the 5' end of the sequence 3 are open reading frame...

Embodiment 2

[0080] Example 2. Expression analysis and subcellular localization of HbSCaBP10 and HbSOS1

[0081] 1. Expression analysis

[0082] 1. Take wild barley seeds, put them in deionized water, soak them at room temperature for 12 hours, and then put them in a refrigerator at 4°C overnight.

[0083] 2. Take the seeds treated in step 1, wash them three times with sterilized deionized water, place them in a petri dish with moist gauze, and germinate at 25°C. After 4-5 days, after most of the seeds germinate, transfer to a sterilized glass bottle filled with 1 / 2 Hoagland culture solution (the glass bottle is wrapped with opaque paper) for cultivation (temperature 22-23°C, light intensity 2000μmol m -2 the s -1 , light 12h / dark 12h), when the seedling grows to two leaves and one heart, take materials.

[0084] 3. The seedlings grown to two leaves and one heart in step 2 are grouped as follows:

[0085] Group I: the seedlings were placed in 1 / 2 Hoagland culture solution containing 350...

Embodiment 3

[0105] Example 3, Application of HbSCaBP10 in Regulating Plant Salt Tolerance

[0106] 1. Obtaining of transgenic strains

[0107] 1. pGreen0029-HbSCaBP10 overexpression vector: insert the double-stranded DNA molecule shown in the sequence 3 from the 5' end nucleotides 1-783 of the sequence listing between the BamHI and MfeI restriction sites of the pGreen0029 vector to obtain pGreen0029 -HbSCaBP10 overexpression vector (sequencing verification is correct).

[0108] 2. The pGreen0029-HbSCaBP10 overexpression vector prepared in step 1 was introduced into Agrobacterium GV3101 to obtain recombinant Agrobacterium HbSCaBP10.

[0109] 3. Use the flower dipping method (refer to the literature: Clough S.J.&BentA.F., 1998. Floral dip: asimplified method for Agrobacterium-mediated transformation of Arabidopsisthaliana. The Plant Journal16, 735-743) with the recombinant Agrobacterium HbSCaBP10 obtained in step 2 Infection of the Arabidopsis mutant sos3 yielded T 0 generation seeds. S...

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Abstract

The invention discloses a protein. The protein is acquired from wild barley, is named as HbSCaBP10 protein as shown in (a1) or (a2): (a1) a protein composed of the amino acid sequence shown in sequence 4 in a sequence list; and (a2) a protein with one or more amino acid residues of the amino acid sequence shown in sequence 4 substituted and / or deleted and / or added, derived from the sequence 4 and related to the biological salt tolerance. The invention also protects an encoding gene (HbSCaBP10 gene) of HbSCaBP10 protein. A test proves that the biological salt tolerance can be promoted by increasing the expression quantity of HbSOS1 gene. The biological salt tolerance can be obviously promoted by increasing the expression quantity of the HbSOS1 gene, the HbSCaBP10 and HbCIPK2 gene. The protein can be applied to the plant breeding and has a positive effect in breeding the salt-tolerance plant.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a plant salt tolerance related protein HbSCaBP10 and its coding gene and application. Background technique [0002] Soil salinization, drought and other adversities are the main environmental limiting factors for plant growth and development. In order to cope with adversity, plants have evolved a set of fine regulatory mechanisms for sensing adversity stress, transmitting adversity signal, and responding at the molecular, cellular and physiological levels during the long-term evolution process. Ca 2+ The dependent signaling pathway plays an important role in a variety of biological processes in plants, including adversity stress responses. In deciphering Ca 2+ In the process of signaling and responding to plant adversity stress, calcium ion receptors are important signal transmitters, regulating the on and off of stress response genes. According to the difference in function and ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07K14/415C12N15/29C12N15/82A01H5/00A01H6/20
CPCC07K14/415C12N15/8273
Inventor 李瑞芬张海纹魏建华
Owner BEIJING ACADEMY OF AGRICULTURE & FORESTRY SCIENCES