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

Method for creating short and strong plant-type chrysanthemums

A technology of chrysanthemum and plant type, applied in chemical instruments and methods, botany equipment and methods, biochemical equipment and methods, etc.

Active Publication Date: 2020-05-26
NANJING AGRICULTURAL UNIVERSITY
View PDF2 Cites 2 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] The object of the present invention is to solve the problem of directional improvement of chrysanthemum plant type, providing a kind of CmYAB3.1 gene, the sequence of this gene is SEQ ID NO.1

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
  • Method for creating short and strong plant-type chrysanthemums
  • Method for creating short and strong plant-type chrysanthemums
  • Method for creating short and strong plant-type chrysanthemums

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0085] Cloning of embodiment 1.CmYAB3.1

[0086] Take cut-flower chrysanthemum 'Shenma' as material, take 0.15 g of leaves, extract the total RNA of the leaves according to the operation method of the Trizol RNA extraction kit (TaKaRa), and perform reverse transcription according to the M-MLV reverse transcription kit (TaKaRa) Obtain cDNA, and use primer 5 software to design specific primers to amplify CmYAB3.1 according to the sequence information of the gene in the chrysanthemum transcriptome database;

[0087] Upstream primer CmYAB3.1-F: 5'-ATGTCTACCATCATCACAGCCAC-3' (SEQ ID NO.2),

[0088] Downstream primer CmYAB3.1-R: 5'-CTAATAATAAGGAGAGACACCAAC-3' (SEQ ID NO.3);

[0089] Using leaf cDNA as template, carry out PCR reaction, 50 μL reaction system: 10×PCR Buffer 5.0 μL, CmYAB3.1-F, CmYAB3.1-R primers 1.0 μL each (20 μmol L -1 ), dNTP mix 4.0μL (2.5mmol·L -1 ), TaqDNA Polymerase 0.2μL, cDNA template 1μL, ddH 2 O 37.8 μL; reaction program: pre-denaturation at 95°C for 3 m...

Embodiment 2

[0090] Example 2. Construction of plant expression vector pORE-R4-35sAA-CmYAB3.1

[0091] Primers were designed according to the full-length gene sequence of CmYAB3.1 for PCR reaction, and restriction sites Xho I and Sma I were introduced into the upstream and downstream of the target gene CmYAB3.1, respectively. Using the plasmid containing the sequence of SEQ ID NO.1 as a template, high-fidelity enzyme (PrimeSTAR TMHS DNA Polymerase, TaKaRa) for PCR reaction, 50 μL reaction system: 10×HS PCRBuffer 5.0 μL, Cm YAB3.1-XhoI-F, Cm YAB3.1-SmaI-R primers 1.0 μL each (20 μmol L -1 ), dNTPmix 4.0μL (2.5mmol·L -1 ), PrimeSTAR TM HS DNA Polymerase 0.4 μL, cDNA template 1 μL, ddH 2 O37.6 μL; reaction program: pre-denaturation at 95°C for 5 min, then melting at 94°C for 30 sec, annealing at 55°C for 30 sec, extension at 72°C for 1 min, 35 cycles of reaction, and extension at 72°C for 10 min; PCR products were recovered using a gel recovery kit (AXYGEN , USA), and then the recovered ...

Embodiment 3

[0094] Embodiment 3. Agrobacterium EHA105 mediates leaf disk method to transform chrysanthemum

[0095] Pick a single colony of EHA105 from a YEB (50 μg / mL rifampicin) plate, inoculate it in 50 mL of YEB liquid medium containing 50 μg / mL rifampicin, cultivate it at 200 rpm, 28°C until the OD value is 0.5, and then bathe the bacteria in ice 30min, centrifuge to collect the bacteria, suspend in 2mL pre-cooled 100mM CaCl 2 (20% glycerol) solution, 200 μL / tube aliquoted for use. Take 10 μL of pORE-R4-35sAA-CmYAB3.1 vector plasmid, add 200 μL of competent cells, bathe in ice for 30 minutes, freeze in liquid nitrogen for 5 minutes, and add 800 μL of YEB liquid medium, pre-culture at 28℃ and 200 rpm for 4 hours. Plate on YEB (50 μg / mL rifampicin + 50 μg / mL kanamycin) solid medium, culture in dark at 28°C for 2 days, pick single clones for detection, and select positive clones to transform chrysanthemums.

[0096] The Agrobacterium used refers to the EHA105 of the plant expression v...

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
Average heightaaaaaaaaaa
Login to View More

Abstract

The invention provides a method for creating a short, small and thick chrysanthemum through a transgenic technology, which belongs to the field of plant genetic engineering and transgenic breeding. The method comprises the following steps: cloning a CmYAB3.1 gene from chrysanthemum; constructing a plant expression vector of the CmYAB3.1 gene; transferring into chrysanthemum by adopting an agrobacterium-mediated method, and culturing to primarily obtain a resistant plant. The molecular detection on the transformed plant proves that the CmYAB3.1 gene is already integrated into the genome DNA ofthe transgenic plant and transcribed. Phenotypic observation and statistical analysis are carried out on transgenic plants, and compared with non-transgenic plants, the height of the transgenic plantsis obviously decreased, and stems are thickened. According to the method, the chrysanthemum plant type is regulated and controlled through the transgenic technology, the short and strong plants suitable for the chrysanthemum morifolium ramat are obtained, the novel and practical method is provided for regulating and controlling the important agronomic traits of the flowers through the genetic engineering technology and easily simplifying cultivation, and the method has high application value.

Description

technical field [0001] The invention belongs to the field of plant genetic engineering and transgenic breeding, and relates to a method for creating dwarf plant-type chrysanthemums through transgenic technology and an application thereof. Background technique [0002] Chrysanthemum (Chrysanthemum morifolium) originated in China and has a long history of cultivation. It is one of the top ten traditional famous flowers in my country and one of the four major cut flowers in the world. The chrysanthemum has the characteristics of rich flower colors and various flower poses, and is deeply loved by people. Plant type directly affects the ornamental quality of flowers, and is one of the main traits that breeders pay attention to. Chrysanthemums can be divided into ground cover chrysanthemums, cut chrysanthemums, potted chrysanthemums, and desk chrysanthemums according to different uses. Breeders regulate the plant type by constantly adjusting the cultivation method, so that it ca...

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): C12N15/29C12N15/82A01H5/00A01H6/14C12Q1/6895C12Q1/686C12N15/11
CPCC07K14/415C12N15/8261C12Q1/686C12Q1/6895C12Q2563/107C12Q2545/114C12Q2521/107
Inventor 陈发棣张雪丁莲李松房伟民胡月姮
Owner NANJING AGRICULTURAL UNIVERSITY
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