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Cs rrm2 gene and application on cotton character improvement

A technology for quality traits and coding genes, applied in the field of plant genetic engineering, can solve the problems such as reports on the application of the domain gene of the rapeseed FCA-γ gene to improve cotton yield or quality traits, etc., and achieve increased fiber length and pollen. Large, the effect of increasing the specific strength at break

Inactive Publication Date: 2012-05-02
INST OF COTTON RES CHINESE ACAD OF AGRI SCI +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] After searching, there is no report on the application of rapeseed FCA-γ gene domain gene in improving cotton yield or quality traits

Method used

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  • Cs rrm2 gene and application on cotton character improvement
  • Cs rrm2 gene and application on cotton character improvement
  • Cs rrm2 gene and application on cotton character improvement

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0058] Embodiment 1: Cloning of Rapeseed Bn-csRRM2 Gene

[0059] 1. Acquisition and purification of Rapeseed cDNA

[0060] (1) Extraction and purification of rapeseed total RNA:

[0061] (a) Grind 0.5 g of the ultra-low temperature frozen rapeseed leaf sample into powder in liquid nitrogen; add RNAiso Plus (TaKaRa Code: D9108A) to completely cover the sample, then let it stand at room temperature until the sample is completely melted, and then continue to grind until The lysate was transparent, and a homogenate was obtained.

[0062] (b) Let the homogenate stand at room temperature for 5 minutes; then centrifuge at 4°C and 12000g for 5 minutes, take the supernatant; add chloroform (1 / 5 volume of RNAiso Plus), cover the centrifuge tube tightly, and vigorously Shake for 15 seconds, wait for the solution to be fully emulsified, then let stand at room temperature for 5 minutes; centrifuge at 4°C and 12000g for 15 minutes, absorb the supernatant, discard the white protein layer i...

Embodiment 2

[0075] Embodiment 2: Construction of csRRM2 gene expression vector

[0076] (1) Recovery of PCR amplification product Use AxyPrep TM DNA Gel Extraction Kit (Cat No.AP-GX-250) Gel Extraction Kit (AXYGEN Company) to recover the electrophoresis of about 300bp in the PCR amplification product in Example 1 Strips, recovery method according to product instructions.

[0077] (2) Link the target fragment to the pGEM-T vector (Promega)

[0078] use -T Easy Vector System I (Cat.#A1360) (Promega, Madison, WI, USA) (the specific operation is carried out according to the product manual)

[0079] Briefly centrifuge the pGEM-T vector to the bottom of the tube. Vortex 2×Rapid Ligation buffer, prepare ligation reaction system in 0.2ml centrifuge tube: 5 μl of 2×Rapid Ligation buffer, 1 μl of pGEM-T Easy Vevtor (50ng), 1 μl of T4 DNA ligase (3u / ml), PCR product (Bn -csRRM2) (25ug / ml) 3 μl, pipet up and down with a gun to mix, and leave overnight at 4°C to obtain plasmid pGEM-T-csRRM2.

[...

Embodiment 3

[0092] Embodiment 3: the acquisition of Bn-csRRM2 gene cotton

[0093] (1) Preparation of the mother solution of MS medium

[0094] Mother solution 1: KNO 3 19000mg / L, NH 4 NO 3 16500mg / L, MgSO 4 .7H 2 O 370mg / L, KH 2 PO 4 1700mg / L, CaCL 2 .2H 2 O 4400mg / L dissolved in 1000mL distilled water, mix well.

[0095] Mother solution 2: MnSO 4 .H 2 O, 1690mg / L, ZnSO 4 .7H 2 O, 860mg / L, H 3 BO 3 620mg / L, KI 83mg / L, Na 2 MO0.5H 2 O 25mg / L, CuSO 4 .5H 2 O 2.5mg / L, CoCl 2 .6H 2 O 2.5mg / L dissolved in 1000ml distilled water, mix well.

[0096] Mother solution 3: Dissolve glycine 200mg / L, thiamine hydrochloride 40mg / L, pyridoxine hydrochloride 50mg / L, nicotinic acid 50mg / L, inositol 10000mg / L in 1000ml distilled water, and mix well.

[0097] Mother liquor 4: Na 2 -EDTA 7.45g / L, FeSO 4 .7H 2 O 5.57g / L, dissolve in 1000ml distilled water, mix well.

[0098] Preparation of MS medium: Dissolve 100ml of mother solution 1, 10ml of mother solution 2, 10ml of mother s...

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Abstract

The invention belongs to the field of plant genetic engineering and discloses a protein relative to crop yield or qualitative character, i.e, a protein of (a) or (b): (a) is composed of an amino acid sequence represented by SEQ ID No.1; (b)is a protein which is formed by an amino acid sequence which is represented by SEQ ID No.1 and is after the deletion, adding or substitution of one or more amino acid residues and is relative to crop yield or qualitative character. The invention also discloses coding genes of the protein. Take the cotton as an example, according to the utility model, the cotyledon, lamina, flowers, bracts, petioles, plant height, cotton boll, etc. can be increased; the fiber length can be increased and the fracture ration strength can be enhanced; the pollen and cell are enlarged; and the transgenic cotton is resistant to premature.

Description

technical field [0001] The invention belongs to the field of plant genetic engineering, and in particular relates to an RNA structural domain gene of rapeseed FCA gene and the application of the structural domain gene in improving economic traits of crops. Background technique [0002] At present, the method of using transgenes to improve crops is mainly based on the known relationship between functional genes and phenotypic traits, using constitutive expression vectors or tissue-specific expression vectors to introduce target genes into target plants in order to obtain specific economic traits or resistance to crops. Improvement of reversible traits. The genes used are all from natural gene sequence with complete structure or cDNA with complete coding sequence. But this method must first know the sequence and function of the gene, so its application is greatly limited. "Method for Transgenically Improving Economic Characters of Crop Using RNA Domain of FCA Gene" (CN200410...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K14/415C12N15/29C12N15/63C12N5/10C12Q1/68A01H1/02A01H5/00
CPCC07K14/415C12N15/8266C12N15/8261Y02A40/146
Inventor 李付广刘传亮武芝侠李凤莲张雪妍商海红张朝军孔德培王倩华王玉芬杨金水孙凡祁巍巍钱晓茵罗小金
Owner INST OF COTTON RES CHINESE ACAD OF AGRI SCI
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