Looking for breakthrough ideas for innovation challenges? Try Patsnap Eureka!

Artemisia carvifolia HD-ZIP IV type transcription factor coding sequence and application

A technology of transcription factor and coding sequence, which is applied to the coding sequence and application field of Qinghao HD-ZIPIV transcription factor, can solve the problem of not regulating glandular hair density and the like

Active Publication Date: 2017-01-04
唐克轩 +1
View PDF4 Cites 9 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In the field of secondary metabolism, there are relatively few reports on secretory glandular hairs
Especially in the study of the traditional Chinese herbal medicine Artemisia annua, the research mainly focuses on the transcriptional regulation of the synthesis key enzyme genes, and there is no report on the regulation of glandular hair density.

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • Artemisia carvifolia HD-ZIP IV type transcription factor coding sequence and application
  • Artemisia carvifolia HD-ZIP IV type transcription factor coding sequence and application
  • Artemisia carvifolia HD-ZIP IV type transcription factor coding sequence and application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0038] Embodiment 1, the cloning of Artemisia annua AaHD1 gene

[0039] 1. Extraction of Total RNA from Artemisia annua Genome

[0040] Take the leaf tissue of Artemisia annua, grind it in liquid nitrogen, add it to a 1.5mL Eppendorf (EP) centrifuge tube filled with lysate, shake it fully, and extract total RNA according to the instructions of the TIANGEN kit. The quality of total RNA was identified by formaldehyde denaturing gel electrophoresis, and then the RNA content was determined on a spectrophotometer.

[0041] 2. Cloning of Artemisia annua AaHD1 gene

[0042]Using the extracted total RNA as a template, cDNA was synthesized under the action of PowerScript reverse transcriptase; gene-specific primers (SEQ ID NO:3 and SEQ ID NO:4) were designed according to the sequence of the AaHD1 gene, and the total cDNA was obtained by PCR. The AaHD1 gene was amplified and sequenced.

[0043] Through the above steps, the full-length coding sequence (SEQ ID NO: 1) of the transcrip...

Embodiment 2

[0044] Example 2, construction of plant binary interference expression vector containing AaHD1 gene

[0045] In order to study the effect of AaHD1 gene on the development of secretory glandular hairs of Artemisia annua, the overexpression vector PHB-AaHD1 overexpressing AaHD1 was constructed. In order to facilitate the construction of the expression vector, the restriction site of BamH1 was introduced into the forward primer, and the restriction site of Xho1 was introduced into the reverse primer. The primers are shown in Table 1;

[0046] Table 1 PCR primers constructed by PHB-AaHD1 vector

[0047]

[0048]

[0049] In this example, the Artemisia annua AaHD1 gene is operably linked to the expression control sequence, and the vector can be used to regulate the content of artemisinin in Artemisia annua through a developmental regulation strategy.

Embodiment 3

[0050] Example 3. Agrobacterium tumefaciens-mediated AaHD1 interference vector genetically transforms Artemisia annua to obtain transgenic Artemisia annua plants

[0051] 1. Acquisition of Agrobacterium tumefaciens Engineering Bacteria Containing AaHD1 Overexpression Vector

[0052] The plant binary overexpression vector containing AaHD1 in Example 2 was transformed into Agrobacterium tumefaciens (such as EHA105, which is a publicly available biological material in the market, which can be purchased from CAMBIA Company in Australia, and the strain number is Gambar 1 ), and validated by PCR. The results showed that the plant binary interference expression vector containing AaHD1 had been successfully constructed into the Agrobacterium tumefaciens strain.

[0053] 2. Agrobacterium tumefaciens mediated AaHD1 gene transformation of Artemisia annua

[0054] 2.1. Preculture of explants

[0055] Artemisia annua seeds were soaked in 75% ethanol for 1 min, then soaked in 20% NaClO...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

PUM

No PUM Login to View More

Abstract

The invention discloses an artemisia carvifolia HD-ZIP IV type transcription factor coding sequence which is marked as AaHD1, the nucleotide sequence of the coding sequence is as shown in SEQ ID NO:1, and the amino acid sequence of the coding sequence is as shown in SEQ ID NO:2. By adopting a transgenic technique, the glandular hair density of the surface of artemisia carvifolia can be effectively regulated and controlled if the artemisia carvifolia is converted by using an artemisia carvifolia AaHD1 transcription factor over-expression vector, so that the content of the artemisia carvifolia can be increased. The glandular hair density of surfaces of leaves of non-transgenic common artemisia carvifolia is 15 pieces per mm<2>, and the glandular hair density of leaves of transgenic artemisia carvifolia expressed by an over-expression AaHD1 gene can be increased to be 25 pieces per mm<2>. Correspondingly, the content of artemisinin can be increased to 13mg / g DW from 9mg / g DW of that of the non-transgenic artemisia carvifolia, and the coding sequence has great significances for providing high-yield and stable new medicine sources for large-scale production of artemisinin.

Description

technical field [0001] The invention relates to the technical field of genetic engineering, in particular to a coding sequence of an Artemisia annua HD-ZIP IV transcription factor and its application. Background technique [0002] Plant metabolism is divided into primary metabolism and secondary metabolism. Primary metabolites (such as sugars, lipids and nucleic acids) exist in all plants and are necessary to maintain cell life activities, while plant secondary metabolites refer to a plant. A large class of small molecular organic compounds that are not necessary for plant growth and development, and their production and distribution are specific to species, tissues and organs, and growth and development. The surface of most plant leaves is not smooth, but has many types of hair-like structures. According to their function and shape, they are divided into two categories: one is cilia without secretory function; the other is glandular hair with secretory function. The main ...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

Application Information

Patent Timeline
no application Login to View More
IPC IPC(8): C12N15/29C07K14/415C12N15/82A01H5/00
CPCC07K14/415C12N15/8205C12N15/8243
Inventor 唐克轩陈明慧颜廷祥沈乾潘琪芳付雪晴
Owner 唐克轩
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Patsnap Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Patsnap Eureka Blog
Learn More
PatSnap group products

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic, Popular Technical Reports.

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap|About US| Contact US: help@patsnap.com