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rh2 protein and its coding gene and application
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A 1.RH2, protein technology, applied in application, genetic engineering, plant genetic improvement and other directions
Active Publication Date: 2021-02-26
THE INST OF BIOTECHNOLOGY OF THE CHINESE ACAD OF AGRI SCI
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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
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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 RH2 protein and its encoding gene and application. The amino acid sequence of the RH2 protein is shown in SEQ ID NO:2. The RH2 gene was introduced into Medicago truncatula R108 to obtain T 0 Medicago truncatula transgenic for the RH2 gene; after self-crossing, the T truncatula 1 Generation of RH2 gene Medicago truncatula. Experiments show that compared with Medicago truncatula R108, T 1 The anthocyanin content in the leaves of Medicago truncatula transgenic with RH2 gene was significantly decreased. RH2 protein and its encoding gene have important theoretical significance and practical value in regulating plant anthocyanin synthesis. The present invention is of great value.
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...
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