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Petal purple spot protein and coding gene thereof

A purpurin and petal technology, applied in genetic engineering, plant genetic improvement, angiosperms/flowering plants, etc., can solve the problem of unclear genetic regulation of sea island cotton petal purpura

Active Publication Date: 2020-06-19
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

[0004] Genetic regulation of petal purpura in sea island cotton is currently unknown

Method used

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  • Petal purple spot protein and coding gene thereof
  • Petal purple spot protein and coding gene thereof
  • Petal purple spot protein and coding gene thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0039] Example 1, Genetic analysis of the purple spot traits at the petal base of sea-island cotton

[0040] (1), test material: upland cotton variety P30A, there is no purple spot at the petal base (hereinafter referred to as "petal purple spot") (see figure 1 ); sea-island cotton variety HaiR (formerly used name: ISR, the applicant self-cultivated sea-island cotton variety) (see figure 1 ), with purple spots at the base of the petals.

[0041] (2) Test method:

[0042] The hybrid F 1 generation; then F 1 Generation selfing, get F 2 generation; during flowering, survey and statistics F 1 Generation and F 2 The number of individual plants with and without purple spots at the base of the petals of the generation.

[0043] Result F 1 All the plants in this generation had purple spots at the base of petals, which indicated that the trait of petal purple spots was controlled by dominant genes. in F 2 A total of 5,540 individual plants were investigated, and the results (...

Embodiment 2

[0046] Example 2 Map-based Cloning of GhBM Gene in Cotton Petal Purple Spot

[0047] (1) test material: the F obtained in embodiment 1 2 Generation segregation population 5540 strains.

[0048] (2) Test method:

[0049] (1) DNA extraction, using DNAsecure Plant Kit (purchased from Tiangen Biochemical Technology (Beijing) Co., Ltd.) and extracting genomic DNA according to the method provided in its manual.

[0050] (2) Initial location of GhBM gene: F 2 The mixed pool of 48 cotton genome DNAs without petal purple spots in the isolated population was used as a template, and the 720 pairs of polymorphic primers between upland cotton and sea island cotton developed by the applicant were used as primers for PCR amplification to carry out preliminary positioning of the GhBM gene .

[0051] The above PCR amplification: the polymorphic primers used (see Table 2) were used for PCR amplification using 2×Taq PCR StarMix with LoadingDye reagent (purchased from Beijing Kangrun Chengye ...

Embodiment 3

[0064] Embodiment 3, cotton petal purpura GhBM gene expression analysis test

[0065] (1) Test materials: flowers of different development stages of upland cotton P30A and sea island cotton HaiR, and different parts of petals.

[0066] (2) Test method:

[0067] 1. RNA extraction:

[0068] RNA was extracted with the EASYspin Plant Ultrapure RNA Rapid Extraction Kit (purchased from Yuanpinghao (Beijing) Biotechnology Co., Ltd.). The specific operation is as follows:

[0069] a) Add 500 μl Lysis Solution RLT (with β-mercaptoethanol added) to a 2ml sterile RNA centrifuge tube, then add 50 μl PLANTaid, mix well and set aside;

[0070] b) Take 100 mg of cotton P30A or HaiR petal tissue, grind it into powder with liquid nitrogen, add 550 μl of the prepared lysate, immediately vortex for 1 min, and let stand at room temperature for 10 min to fully lyse;

[0071] c) Centrifuge at 12000rpm for 10min at 4°C, take 500μl supernatant into a new 1.5ml RNase-free centrifuge tube;

[0072...

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Abstract

The invention discloses a pedal purple spot protein which is composed of an amid acid sequence which is represented by SEQ ID No.1. Furthermore the invention discloses a gene which is composed of thenucleotide sequence that is represented by the SEQ ID No.2 for coding the protein. The invention further discloses application of the protein and the coding gene thereof in regulating the petal purplespot characteristic of the plant. The invention defines a fact that the petal purple spot characteristic is controlled by a single dominant gene and can be used as a marking characteristic of a sea-land hybrid. The gene can be transplanted into a flower plant through a transgene method, thereby supplying a new gene source for abundance of the flower plants.

Description

technical field [0001] The invention belongs to the technical field of plant genetic engineering, and specifically relates to a petal purpura protein, and also relates to a coding gene of the petal purpura protein and an application thereof. Background technique [0002] The heterochromatic spots and stripes on the flowers, leaves, fruits and branches of plants are collectively referred to as colorful spots, especially the colorful spots on the petals, which are the main components of the ornamental value of horticultural plants. According to the size, shape and position of the flower spots on the petals, the color spots on the petals are divided into two categories: regular color spots and irregular color spots. Regular color spots include flower rings, flower centers (flower eyes), flower spots, flower ribs and Lace and other forms; irregular color spots refer to non-fixed patterns of different color scattered points or stripes on the petals. Regular color spots are usual...

Claims

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Application Information

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IPC IPC(8): C07K14/415C12N15/29C12N15/82A01H5/02A01H6/60
CPCC07K14/415C12N15/8218C12N15/825
Inventor 梁成真穆罕默德·阿里·阿比德孟志刚王远张锐
Owner THE INST OF BIOTECHNOLOGY OF THE CHINESE ACAD OF AGRI SCI
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