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RH2 protein as well as coding gene and application thereof

1. RH2, protein technology, applied in application, genetic engineering, plant genetic improvement, etc.

Active Publication Date: 2020-05-15
THE INST OF BIOTECHNOLOGY OF THE CHINESE ACAD OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

With the in-depth development of functional genomics of Medicago truncatula, some genes related to anthocyanin biosynthesis pathway and MYB, bHLH and WD40 family transcription factors were cloned in Medicago truncatula. Metabolic regulation plays an important role, but due to the special physiological and structural characteristics of legumes, the expression patterns and biological functions of the above genes are different from those reported in the dicotyledonous model plant Arabidopsis, indicating that anthocyanins in different species There is genetic diversity in synthetic and metabolic regulatory pathways

Method used

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  • RH2 protein as well as coding gene and application thereof
  • RH2 protein as well as coding gene and application thereof
  • RH2 protein as well as coding gene and application thereof

Examples

Experimental program
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Effect test

Embodiment 1

[0085] Cloning of embodiment 1, RH2 gene

[0086] 1. Phenotype analysis of mutant rh2-1 and mutant rh2-2

[0087] Mutant rh2-1 and mutant rh2-2 were both inserted by Medicago truncatula Tnt1 from The Nobel Foundation (https: / / www.nobelprize.org / the-nobel-prize-organisation / the-nobel-foundation / ) Provided by the mutant library, the numbers of mutant rh2-1 and mutant rh2-2 in the Tnt1 insertion mutant library of Medicago truncatula are NF21135 and NF21515 in sequence. According to the information recorded in the Medicago truncatula Tnt1 insertion mutant library, the mutant rh2-1 is a Medicago truncatula mutant obtained by inserting Tnt1 into the second exon of the RH2 gene, and mutant rh2-2 is the third exon of the RH2 gene inserted into Tnt1. Mutants of Medicago truncatula obtained on exons.

[0088] The mutant rh2-1, mutant rh2-2 and Medicago truncatula R108 were planted respectively, and the leaf phenotypes were observed.

[0089] Leaf phenotype see figure 1 (A is the bl...

Embodiment 2

[0108] Example 2, T 1 Obtaining, Phenotype Identification and Detection of Anthocyanin Content of Medicago truncatula Transduced with RH2 Gene

[0109] 1. Construction of recombinant plasmid 35S::RH2

[0110] 1. The PCR amplification product obtained in Step 3 of Example 1 and the vector pDONR207 (Invitrogen) were subjected to BP reaction to obtain an intermediate vector.

[0111] 2. After completing step 1, perform LR reaction with the intermediate vector and pEarleyGate202 plasmid to obtain the recombinant plasmid 35S::RH2.

[0112] 3. Sequencing the recombinant plasmid 35S::RH2.

[0113] Sequencing results showed that the recombinant plasmid 35S::RH2 contained the DNA molecule shown in SEQ ID NO: 1 (ie the RH2 gene), and the expression of the RH2 gene was driven by the 35S promoter.

[0114] Second, the acquisition of recombinant Agrobacterium

[0115] The recombinant plasmid 35S::RH2 was introduced into Agrobacterium tumefaciens AGL1 to obtain the recombinant Agrobacte...

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Abstract

The invention discloses an RH2 protein as well as a coding gene and application thereof. An amino acid sequence of the RH2 protein is represented by SEQ ID NO:2. The RH2 gene is introduced into medicago truncatula R108, so as to obtain T0-generation simulated-converted R2H2 gene medicago truncatula; and T1-generation converted R2H2 gene medicago truncatula is obtained through selfing. Experimentsprove that compared with the medicago truncatula R108, the content of anthocyanin in leaves of the T1-generation converted R2H2 gene medicago truncatula is remarkably decreased. The RH2 protein and the coding gene of the RH2 protein have important theoretical significances and practical values to the regulation and control of synthesis of plant anthocyanin.

Description

technical field [0001] The invention belongs to the field of biotechnology, and specifically relates to RH2 protein and its coding gene and application. Background technique [0002] Anthocyanins are an important class of flavonoids, which widely exist in the flowers, fruits, seeds and vegetative tissues of plants, making the tissues appear in different colors from orange, red, purple to blue. Anthocyanins usually form spots, stripes, and vein-like specific enrichment patterns on reproductive organs such as flowers and fruits, and have important ecological functions in attracting insects to pollinate, animals to feed on, and maintaining the reproduction of plant progeny. Compared with reproductive organs such as flowers and fruits, the accumulation of anthocyanins on leaves can protect plants from biotic and abiotic stress; however, specific anthocyanin accumulation on leaves with leaf scars is relatively rare, and generally occurs in clover and Medicago truncatula , can be...

Claims

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Application Information

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IPC IPC(8): C07K14/415C07K19/00C12N15/29C12N15/62C12N15/82A01H6/54
CPCC07K14/415C07K2319/21C07K2319/22C07K2319/41C07K2319/43C12N15/825
Inventor 林浩王崇楠牛丽芳
Owner THE INST OF BIOTECHNOLOGY OF THE CHINESE ACAD OF AGRI SCI
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