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Panax ginseng cle family peptides and their application in the regulation of plant root growth and development

A plant and polypeptide technology, applied in the fields of application, plant peptides, plant products, etc., can solve the problems of ginseng CLE family peptide regulation and other problems that have not been seen, and achieve the effects of promoting primary root differentiation, inhibiting primary root growth, and good research potential

Active Publication Date: 2021-10-01
INST OF CHINESE MATERIA MEDICA CHINA ACAD OF CHINESE MEDICAL SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

But so far there have been no reports about ginseng CLE genes and CLE polypeptides, nor have we seen any possible regulatory effects of ginseng CLE family polypeptides on plants

Method used

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  • Panax ginseng cle family peptides and their application in the regulation of plant root growth and development
  • Panax ginseng cle family peptides and their application in the regulation of plant root growth and development
  • Panax ginseng cle family peptides and their application in the regulation of plant root growth and development

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0052] Example 1, 4 people found in Clear polypeptide

[0053] First, get 4 ginseng CLE genes

[0054] Using the local Local Blast method, the sequence of similarity in the ginsence Cle1-CLE45 gene sequence (Expection Value = 0.001) is used to filter the CLE sequence of specific expression in roots, and use ORFFINDER software to find complete The gene sequence of the ORF frame is carried out in PCR primer design.

[0055] The sample was pulverized with liquid nitrogen by liquid nitrogen, and total RNA was extracted with Trizol method, and RNA integrity was determined using 1% agarose gel electrophoresis, and the nucleic acid / protein quantitator was measured. CDNA was synthesized using a Takara anti-transcription kit method (M-mlvrtase cDNA Synthesis Kit). The PCR was obtained by cDNA as a template, and PCR amplification was performed using primers to obtain PCR products.

[0056] Second, 4 ginseng CLE gene expression analysis

[0057] The nucleic acid sequence obtained by step i...

Embodiment 2

[0070] Example 2, a ginseng CLE polypeptide can regulate the long and root branch of the Arabidopsis root

[0071] Test polypeptide: PgCLE12 polypeptide, PgCLE25 polypeptide, PgCLE27 polypeptide, PgCLE45 polypeptide, PgCLE3 PgCLE5 polypeptide or polypeptide.

[0072] 1, Arabidopsis thaliana seeds ecotype Columbia take, with 75% aqueous ethanol for 3min, then with sterile deionized water 10 times.

[0073] 2, the packet processing:

[0074] The first group (Group Mock): Step 1 is taken to complete the seed, were sown in 1 / 2MS media plates, 22 ± 2 ℃, light 9 days;

[0075] The second group (PgCLE3 group): Step 1 is taken to complete the seed sown containing 1 / 2MS medium plates 1μM PgCLE3 polypeptide, 22 ± 2 ℃, light 9 days;

[0076] The third group (PgCLE5 group): Step 1 is taken to complete the seed sown containing 1 / 2MS medium plates 1μM PgCLE5 polypeptide, 22 ± 2 ℃, light 9 days;

[0077] The fourth group (PgCLE12 group): Step 1 takes to complete the seeds were sown in 1 / 2...

Embodiment 3

[0085] Example 3, CLE polypeptide ginseng can promote differentiation of primary roots Arabidopsis

[0086] Example 3 after the completion of step 2, the primary root of the plant taken, using two-photon laser scanning confocal microscope LSM880 of the primary root meristem region were observed and photographed on the meristematic region length is detected.

[0087] See a photo figure 2 (A), the length of the primary root meristem region average see figure 2 (b).

[0088] Compared with Mock group, PgCLE3 polypeptide and PgCLE5 polypeptide had no effect on the primary root differentiation, PgCLE45 polypeptide, PgCLE12 polypeptide, PgCLE25 polypeptide and PgCLE27 polypeptide can be significantly inhibited the length of the primary root meristem region, to promote the maturation zone earlier formed, especially PgCLE45 promoting differentiation polypeptide of the most obvious.

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Abstract

The invention discloses a ginseng CLE family polypeptide and its application in the regulation of plant root growth and development. Polypeptides provided by the present invention: (a) PgCLE45 polypeptide: as shown in sequence 1 of the sequence listing; (b) PgCLE12 polypeptide: as shown in sequence 2 of the sequence listing; (c) PgCLE25 polypeptide: as shown in sequence 3 of the sequence listing ; (d) PgCLE27 polypeptide: as shown in sequence 4 of the sequence listing. The present invention also protects the application of any of the above polypeptides in inhibiting plant root tip cell proliferation, in promoting plant root differentiation, in reducing the length of plant root meristems, in reducing plant Use in root length, use in reducing the number of root branches in plants. As a plant polypeptide hormone, ginseng CLE polypeptide has good research potential and broad application prospects.

Description

Technical field [0001] The invention belongs to the field of biotechnology, involving ginseng CLE family polypeptides and its application in the growth and development of plant roots. Background technique [0002] CLE (CLAVATA3 / EMBRYO SURROUNDING REGEN) is the hottest plant polypeptide in the past decade, and it is also the largest plant polypeptide molecular family to date, mainly by encoding a small secreted protein to participate in the signal transduction between cells, and in the plant Maintain balance between stem cell proliferation and differentiation during growth and development. So far, the CLE polypeptide has been found to play a key role in the formation of seed development, vascular beam formation, side root growth, stem top and root end of the stem cell division and differentiation. The CLE polypeptide has a deeper study in Arabidopsis with rice, and the study of CLE polypeptide hormone in the pharmaceutical plant is blank. [0003] People participating in the fiv...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07K14/415C12N15/29C12N15/82A01H5/06A01H6/00A01H6/20
CPCC07K14/415C12N15/8261
Inventor 黄璐琦刘娟蒋超袁媛
Owner INST OF CHINESE MATERIA MEDICA CHINA ACAD OF CHINESE MEDICAL SCI
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