Medicago truncatula multi-petal control protein and encoded genes and application thereof

A gene and coding technology, which is applied to the multi-petal control protein of Medicago truncatula and its encoded gene and application field, can solve the problems of protein functional domain deletion, complex regulation of butterfly flower and flower organ development, etc.

Active Publication Date: 2018-06-22
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

However, due to the specificity of the shape and development of Papillon flower organs, as well as the gene doubling, gene functional differentiation, gene expression pattern changes, and protein functional domain deletion or acquisition in Papilionaceae plants, the Papilionaceae Regulation of floral organ development becomes more complex

Method used

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  • Medicago truncatula multi-petal control protein and encoded genes and application thereof
  • Medicago truncatula multi-petal control protein and encoded genes and application thereof

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Experimental program
Comparison scheme
Effect test

Embodiment 1p

[0034] The acquisition and phenotype analysis of embodiment 1pis mutant

[0035] By screening the Tnt1 insertion mutant library of Medicago truncatula, a mutant NF10617 with increased petals and complete loss of stamens and pistils was obtained, named Petals Increased Severely (pis).

[0036] Detailed phenotypic observations of pis mutants such as figure 1 shown. figure 1 Middle A is the side view of the wild-type flower bud at the floret stage; B and C are the side view and top view of the wild-type young flower, respectively; D and E are the side view and top view of the wild-type mature flower, respectively; F is the pis mutant small Side view of flower bud; G and H are side view and top view of young flower of pis mutant respectively; I and J are side view and top view of mature flower of pis mutant In addition to the increase, there are also calyxes). Compared with the wild type, mutant pis exhibited enlarged buds throughout the flower development period. Figure K sho...

Embodiment 2

[0038] The cloning of embodiment 2PIS gene

[0039] According to the Tnt1 website ( NF10617 published at https: / / medicago-mutant.noble.org / mutant / The flanking sequence, design the corresponding primers,

[0040] NF10617-F:5'-CCTCCTCTAACCTGCTCCA-3';

[0041] NF10617-R:5'-TCACCACCTCTTTCCCATTCA-3';

[0042] Detect linkage of mutants to genes. Finally, we found a gene PIS linked to the mutant, DNA level detection, compared with the wild type, the insertion site of the mutant amplified a band about 5.5K longer than the wild type ( figure 2 B), determined to be Tnt1 sequence by sequencing comparison. In the NF10617 mutant, Tnt1 was inserted in the first exon of the gene. In order to prove that the PIS gene is the control gene of the mutant, we ordered two other mutants of this gene: NF21657 (pis-2 inserted in the first exon), NF16829 (pis-3 inserted in the fourth exon exon) and named the previous NF10617 as pis-1 (see figure 2 A). Observing the phenotypes of the homozygo...

Embodiment 3PI

[0043] The acquisition of embodiment 3PIS gene and its coded protein PIS

[0044] 1. Extract the RNA of the blooming flower of Medicago truncatula R108 (wild type), and reverse transcribe it into cDNA.

[0045] 2. Using the cDNA obtained in step 1 as a template, amplify using a primer pair consisting of primer PIS-F and primer PIS-R to obtain a PCR amplification product.

[0046] PIS-F: 5'-CACCATGGTTGGTGACAAAGGAGA-3';

[0047]PIS-R: 5'-TCACATCAAACCACCACCAC-3'.

[0048] 3. Sequence the PCR amplification product obtained in step 2 to obtain the coding region sequence of the target gene, as shown in sequence 1 in the sequence listing, and encode the protein shown in sequence 2 in the sequence listing. Consists of 1009 amino acid residues. The CDS of the gene was compared on the Medicago website (http: / / blast.jcvi.org / Medicago-Blast / ), and the genome sequence of the PIS gene was obtained, as shown in Sequence 3. The protein shown in Sequence 2 of the Sequence Listing is named ...

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Abstract

The invention discloses a medicago truncatula multi-petal control protein and encoded genes and application thereof. The medicago truncatula multi-petal control protein is shown as the following (a1)or (a2), wherein (a1) is a protein consisting of the amino acid sequence shown as SEQ ID NO.2, and (a2) is a protein which is derived from SEQ ID NO.2 through substitution and / or deletion and / or addition of one or more amino acid residues of the amino acid sequence shown as SEQ ID NO.2 and has functions the same as the protein of (a1). The genes can control the increase of petals of medicago truncatula, finite growth of plants is achieved, mutation of the leaf morphology is achieved, and the petiole gets shorter.

Description

technical field [0001] The invention belongs to the technical field of genetic engineering, and in particular relates to a multi-petal control protein of Medicago truncatula and its coded gene and application. Background technique [0002] Flowers are the unique reproductive organs of angiosperms. The normal development of flower organs is directly related to the reproduction of species, and also affects the yield, quality, use value and economic benefits of flowering crops. In addition, flower size, morphological structure, and the number of constituent organs are the most commonly used morphological traits in the study of systematic and evolutionary botany. The study of floral organ development is also of great significance to the study of the origin of species and evolution. [0003] Medicago truncatula (Medicago truncatula) belongs to the Fabaceae Papilionaceae subfamily, and its flowers are typical bilaterally symmetrical Papilionaceae. In recent years, with the comple...

Claims

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Application Information

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IPC IPC(8): C07K14/415C12N15/29C12N15/82A01H5/00A01H6/54
CPCC07K14/415C12N15/827
Inventor 林浩祝步拓李辉牛丽芳
Owner THE INST OF BIOTECHNOLOGY OF THE CHINESE ACAD OF AGRI SCI
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