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

Genetic transformation method for sedum plant

A genetic transformation method, the technology of Sedum genus, applied in biochemical equipment and methods, using vectors to introduce foreign genetic material, recombinant DNA technology, etc., can solve the problems of not having hyperaccumulation and high resistance, and achieve major applications Promote the effect of high value and high genetic transformation rate

Active Publication Date: 2017-06-09
INST OF BOTANY CHINESE ACAD OF SCI
View PDF5 Cites 1 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Moreover, Sedum sedum with ore only has high resistance and super-accumulation ability to cadmium and zinc, but it does not have super-accumulation and high resistance to other heavy metals such as arsenic, copper, mercury, etc.

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
  • Genetic transformation method for sedum plant
  • Genetic transformation method for sedum plant
  • Genetic transformation method for sedum plant

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0068] Embodiment 1, the optimization of clustering bud induction parameter

[0069] Use the stem segment of Sedum sedum as explants to induce clustered buds, and the specific steps are as follows:

[0070] 1. To collect fresh stems of Sedum sedum, follow the steps below:

[0071] Rinse with tap water→disinfect with 75% (volume ratio) alcohol aqueous solution for 30s→rinse with sterile water for 3 times and blot dry with sterile filter paper→use 0.1g / 100mL HgCl 2 Soak in aqueous solution for 6 minutes (shake gently during the period) → rinse with sterile water at least 4 times and blot dry with sterile filter paper.

[0072] 2. After step 1 is completed, the stem is longitudinally cut into stem sections with a length of about 1 cm (each stem section has 1 stem node).

[0073] 3. Get the stem section obtained in step 2, place it on the bud induction medium and cultivate it for 3 weeks (subculture once a week, all adopt the same bud induction medium; culture conditions: 25°C, ...

Embodiment 2

[0079] Embodiment 2, optimization of screening parameters

[0080] 1. Same as step 1 in Example 1.

[0081] 2. Same as step 2 of embodiment 1.

[0082] 3. Take the stem segment obtained in step 2 and place it on the bud induction medium for 3 weeks (subculture once a week, all using the bud induction medium; culture conditions: 25° C., 14 hours of light / 10 hours of darkness).

[0083] The bud induction medium is MS solid medium (pH5.8) containing 0.1 mg / L 6-BA and 1.0 mg / L NAA.

[0084] 4. After completing step 3, take the stem segment with clustered buds, cut off the leaves and place it on the screening medium for 3 weeks (subculture once a week, all using the same screening medium; culture conditions: 25 ° C, 14 hours of light / 10 hours of darkness).

[0085] The screening medium was MS solid medium (pH5.8) containing antibiotics, 0.1 mg / L 6-BA and 1.0 mg / L NAA. When the antibiotic is hygromycin (Hyg), the concentration in the selection medium is 5 mg / L, 10 mg / L, 20 mg / L...

Embodiment 3

[0089] Embodiment 3, genetic transformation

[0090] The plant expression vector used is pSN1301 plasmid. The pSN1301 plasmid is transformed on the basis of the pCAMBIA1301 plasmid, and has a GUS reporter gene driven by the CaMV 35S promoter. The literature that records the pSN1301 plasmid is: Guo, J.; Dai, X.; Xu, W.; Ma, M. Overexpressing GSH1 and AsPCS1 simultaneously increases the tolerance and accumulation of cadmium and arsenic in Arabidopsis thaliana. Chemosphere 2008, 72, 1020-1026 .). The schematic diagram of the elements of the pSN1301 plasmid is shown in Figure 4 . Figure 4 Middle: LB represents T-DNA insertion left border; RB represents T-DNA insertion right border; HPT represents hygromycin phosphotransferase gene; 35SP represents CaMV35 promoter; MCS represents multiple cloning site; GUS represents β-glucoside acidase gene.

[0091] 1. Preparation of infection solution

[0092] 1. Introduce the pSN1301 plasmid into Agrobacterium tumefaciens C58 to obtain ...

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

PropertyMeasurementUnit
lengthaaaaaaaaaa
Login to View More

Abstract

The invention discloses a genetic transformation method for a sedum plant. The invention firstly discloses a genetic transformation method for the sedum plant. The genetic transformation method comprises the following steps: (1) taking a stem fragment of the sedum plant, inducing the stem fragment to form cluster buds, and obtaining a meristem block; (2) carrying out co-culture on the meristem block obtained in the step (1) and recombined agrobacterium, wherein the recombined agrobacterium is obtained by introducing a plasmid with a target DNA molecule into start agrobacterium; (3) inducing the meristem block obtained in the step (2) for germination; (4) inducing the germinated meristem block for rooting. The method provided by the invention has the advantage of high genetic transformation rate, and has an important application and popularization value for genetic transformation of the sedum plant and gene engineering researches.

Description

technical field [0001] The invention relates to a method for genetic transformation of sedum plants. Background technique [0002] Cadmium (Cd) has serious toxic effects on all organisms. At present, my country's soil cadmium pollution is serious, and there is a trend of increasing year by year. Cadmium in the soil has a strong migration ability, is easily absorbed by plants, and enters the human body through the food chain, seriously endangering people's health. Phytoremediation is an ideal way to control soil heavy metal pollution such as cadmium. In recent years, more cadmium hyperaccumulator plants have been studied, including Arabidopsis thaliana, Sedum chinensis, Sedum sedum and Sedum sedum. Cadmium hyperaccumulator plants are also zinc hyperaccumulator plants. Compared with other cadmium hyperaccumulator plants, Sedum sedum with ore has strong cadmium enrichment ability and large biomass (Cao D, Zhang HZ, Wang YD, Zheng LN(2014) Accumulation and distribution chara...

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
Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/82
CPCC12N15/8205
Inventor 徐文忠刘桓赵海霞
Owner INST OF BOTANY CHINESE ACAD OF SCI
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