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

Method for identifying two parents of nymphaea tetragona hybrid based on ITS sequence and matK sequence

A sequence and water lily technology, applied in the field of plant molecular identification, can solve the problems of unidentified water lily hybrids

Active Publication Date: 2022-07-22
KUNMING INST OF BOTANY - CHINESE ACAD OF SCI
View PDF1 Cites 0 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] At present, there is no report on the method of identifying two parents of water lily hybrids based on ITS sequence and matK sequence in the prior art

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 identifying two parents of nymphaea tetragona hybrid based on ITS sequence and matK sequence
  • Method for identifying two parents of nymphaea tetragona hybrid based on ITS sequence and matK sequence
  • Method for identifying two parents of nymphaea tetragona hybrid based on ITS sequence and matK sequence

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0053] This embodiment provides a method for constructing a database of Nymphaea ITS and matK nucleotide sequences, and the method can update the constructed nucleotide sequence database after adding online public databases or newly sequenced sequences. Specifically include the following steps:

[0054] 1. Download the existing Nymphaea ITS and matK nucleotide sequences. Search the National Center for Biotechnology Information with the syntax "((Nymphaea[Organism])AND5.8S[Title])NOT PREDICTED[Title]" and "((Nymphaea[Organism])AND matK[Title])NOTPREDICTED[Title]" (NCBI) Nucleic Acid Database ( www.ncbi.nlm.nih.gov / nucleotide ) to obtain all ITS and matK sequence data of Nymphaea. As a result, 499 ITS sequences were obtained, and the sequence length ranged from 213bp to 1002bp. A total of 49 matK sequences were obtained. After removing two unverified sequences (KJ747597 and JQ024977), 47 remained. The lengths of the 47 matK sequences ranged from 556bp to 2585bp. In order to...

Embodiment 2

[0070] The present embodiment provides a method for obtaining the ITS (monoclonal) and matK nucleotide sequences of the species of the Nymphaea genus to be determined, which specifically includes the following steps:

[0071] 1. Extract the genomic DNA of the water lily of the parent to be determined. Total DNA from water lily leaves was extracted using a modified 2×CTAB method (ROGERS and BENDICH, 1985). The general process includes grinding under liquid nitrogen, 65°C water bath of CTAB extract, chloroform isoamyl alcohol extraction, isoamyl alcohol precipitation, ethanol washing, ribonuclease digestion, etc.

[0072] 2. Design primers. Download the 18S and 25S sequence data of Nymphaea from the NCBI nucleotide database, combine with the ITS sequence data downloaded in Example 1, search for the ITS primer pairs commonly used in plants, and find the product size is suitable, and the Nymphaea 18S and 25S sequences are found. The corresponding primers with high complementarit...

Embodiment 3

[0077] Using the leaves of a water lily variety, N. 'Joey Tomocik', planted in the Botanical Garden of the Kunming Institute of Botany, Chinese Academy of Sciences, as the material, DNA extraction, PCR amplification and sequencing were carried out using the method in Example 2 to obtain a single clone of the water lily. There are 10 ITS sequences of 10 "genotype" sequences, and the sequence length is 855bp ~ 935bp (see figure 1 , figure 2 , image 3 ), two matK sequences were obtained, with sequence lengths of 977bp and 1008bp (see Figure 4 , Figure 5 ).

[0078] The ITS sequences (10) of Huang Joey's water lily monoclonal were aligned with the Nymphaea ITS nucleotide sequence database constructed in Example 1, and a neighbor-joining tree was constructed. and fragrant water lily (N.odorata, including its two varieties subsp.odorata and subsp.tuberosa) into a clade (see figure 1 ). Extract the branch sequence and then compare and build an adjacency-joining tree. The ...

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 provides a method for identifying two parents of a nymphaea tetragona hybrid based on an ITS sequence and a matK sequence, and belongs to the technical field of plant molecular identification. The method comprises the following steps: obtaining a nuclear genome fragment ITS sequence of parent heredity and a chloroplast genome fragment matK sequence of maternal heredity of the nymphaea tetragona hybrid based on first-generation sequencing, and carrying out one-generation sequencing on the ITS and matK sequences downloaded by GenBank to obtain two parents of the nymphaea tetragona hybrid; the method comprises the following steps: respectively establishing ITS and matK sequence databases of water lily after strict screening, respectively comparing ITS and matK sequences of a parent water lily to be determined with the corresponding databases, constructing an adjacency tree based on genetic distance, and checking specific sites to obtain information of a male parent and a female parent of the water lily to be determined. Results show that based on a water lily ITS and matK nucleotide sequence database downloaded and constructed based on GenBank, by sequencing the ITS sequence and matK sequence of the water lily hybrid of the parent to be identified, the female parent and male parent information of the water lily hybrid can be identified through a genetic distance method.

Description

technical field [0001] The invention belongs to the technical field of plant molecular identification, and in particular relates to a method for identifying two parents of a water lily hybrid based on an ITS sequence and a matK sequence, a combination of a nuclear DNA fragment ITS nucleotide sequence and a chloroplast DNA fragment matK nucleotide sequence pair Methods for molecular identification of water lily hybrids (Nymphaea hybrid - natural or artificial hybrid, see Chapter I Article 1 of the Ninth Edition of the International Code of Nomenclature of Cultivated Plants, 2016). Background technique [0002] Nymphaea is a general term for plants of the Nymphaea genus (Nymphaea), which are widely distributed all over the world. The genus Nymphaea can be divided into 5 subgenera, and can also be divided into cold-resistant Nymphaea and tropical Nymphaea according to the ecological and ecological adaptability. There are many varieties and rich colors of water lilies, known as...

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): A01H1/02A01H1/04C12Q1/6895C12Q1/6869C12N15/11G16B20/30G16B50/00
CPCA01H1/021A01H1/04C12Q1/6895C12Q1/6869G16B50/00G16B20/30C12Q2531/113C12Q2563/185C12Q2565/125G16B30/10
Inventor 贺正山杨俊波张志荣杨继雄杨静贾艳霞曾春霞
Owner KUNMING 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