Anthurium soc transcription factor aasoc and its coding gene and application

A technology of transcription factors and coding genes, which is applied in the fields of application, genetic engineering, plant gene improvement, etc., can solve the problems that transcription factors have not yet been discovered, and achieve the effect of promoting early flowering

Active Publication Date: 2018-08-17
ZHEJIANG XIAOSHAN COTTON & FLAX RES INST
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Anthurium molecular breeding has just started, and many transcription factors regulating flowering have not yet been discovered

Method used

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  • Anthurium soc transcription factor aasoc and its coding gene and application
  • Anthurium soc transcription factor aasoc and its coding gene and application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0026] Example 1 Obtaining the transcription factor AaSOC derived from Anthurium

[0027] In August 2014, the young bracts and leaves of the 6-year-old Anthurium variety "Alabama" were taken as materials, and the total RNA was extracted with the Trizol reagent produced by Invitrogen Company, and then the total RNA was purified using the reverse transcription kit produced by Promega Company. RNA is processed and reverse-transcribed to synthesize cDNA, which is used as a template. Using the homologous cloning method, design the conservative degenerate primer pair P1-TCTCCCCGCGCGGCAAGC (SEQ ID NO: 3), P2-GAATAATTGATTCTTCT (SEQ ID NO: 4), and use the above template to amplify, the reaction program is 95 ℃ for 1 min , 94°C for 50 s, 57°C for 30 s, 72°C for 45 s, 36 cycles, 72°C for 5 min, the reagents (LATaq enzyme) used were purchased from Dalian Bao Biological Company, the amplification system: LAtaq enzyme (5 U / µl ) 0.5 µl, 10×LAtaq buffer Ⅱ 5 µl, dNTP mixture 8 µl, template 1 ...

Embodiment 2

[0035] Embodiment 2 Preparation of recombinant genetically engineered bacteria containing transcription factor AaSOC coding gene

[0036] Using the specific primer pair: GSPf--ctggtaccATGGTGAGGGGGAAGACA (SEQ ID NO: 8, containing the Kpn1 restriction site), GSPr-gctctagaTCATTTTTCTCTGCCTTGGGT (SEQ ID NO: 9, containing the Xba1 restriction site), the high The cDNA of the first-strand product of reverse transcription was used as a template for PCR reaction. The system refers to the above-mentioned RACE reaction system. The reaction program is: 95 °C for 1 min, 94 °C for 50 s, 66 °C for 1 min, 72 °C for 1.5 min, 36 cycles, 72°C for 5 min. The obtained PCR product was recovered and subjected to Kpn1 and Xba1 double enzyme digestion, and the enzyme digestion system was carried out according to the restriction enzyme specification of TAKARA company. The gene fragment after enzyme digestion and the expression vector pCAMBIA2300 (M) recovered by the same enzyme digestion (the carrier i...

Embodiment 3

[0037] Example 3 Application of gene encoding transcription factor AaSOC in the preparation of very early flowering plants

[0038] The Agrobacterium strain EHA105 containing the overexpression vector prepared in Example 2 was used to infect the leaves of common tobacco aseptic seedlings, and the general leaf disk method was used to transform (materials and methods strictly refer to Horsch RB, Fry JE, Hoffman NL, Eicholtz D, Rogers SG, Fraley RT (1985) A simple and general method for transferring genes into plants. Science 227:1229–1231). Through differentiation, regeneration and screening, resistant tobacco plants containing transcription factor AaSOC coding gene are obtained. After rooting, they were transplanted into the greenhouse for character identification and analysis.

[0039] After transplanting, the growth of the transgenic resistant plants showed obvious differences from the wild type and the control (empty vector transformed plants), mainly reflected in the short...

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Abstract

The invention discloses an anthurium andraeanum SOC transcription factor AaSOC and an encoding gene and application thereof. The transcription factor AaSOC has the completely-predicted amino acid sequence shown in SEQ ID NO:1 and the encoding gene shown in SEQ ID NO:2. An SOC type transcription factor AaSOC for regulating and controlling the flowering period is cloned from anthurium andraeanum for the first time; by comparing the sequence with the sequence of the same type of SOC transcription factors of different species such as Arabidopsis, it is found that the sequence has conservatism; through transgenic experiments, it is found that the gene has remarkable capacity for promoting the early flowering of converter plants, and it is proved that the factor is a transcription factor with quite high application value. Through the technical scheme, the factor has important significance in studying the flowering mechanism of anthurium andraeanum and culturing a dwarfed and early-flowering species.

Description

technical field [0001] The invention relates to an SOC transcription factor AaSOC derived from Anthurium anthurium, its encoding gene and application method. Background technique [0002] Anthurium ( Anthurium andraeanum ) is a cosmopolitan potted and fresh-cut flower. It occupies a very important position in the cultivation of high-grade potted flowers in my country. However, like other large flowers, my country's anthurium breeding is also at a relatively backward stage, mainly because we do not have enough germplasm resources. [0003] Whether it is hybrid breeding or biotechnology breeding such as transgenic, the shortening of breeding time is self-evident for the value of breeding. Anthurium molecular breeding has just started, and many transcription factors that regulate flowering have not yet been discovered. [0004] SOC transcription factors have a significant function of promoting seedling flowering in different species. In order to better study the function of...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/29C07K14/415C12N15/74C12N15/82A01H5/00A01H6/82
CPCC07K14/415C12N15/743C12N15/827
Inventor 马广莹史小华邹清成金亮朱开元刘慧春詹菁
Owner ZHEJIANG XIAOSHAN COTTON & FLAX RES INST
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