Co-dominance InDel molecular marker for identifying single embryo and multiple embryos of citrus and application thereof

A molecular marker, co-dominant technology, applied in the field of citrus genomics, can solve the problems of laboriousness, low accuracy, slow progress, etc., and achieve the effects of improving breeding efficiency, low cost, and early time.

Active Publication Date: 2016-05-25
HUAZHONG AGRI UNIV
View PDF0 Cites 14 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] The identification of multiple embryos and single embryos is generally observed under a microscope with the naked eye, and there are problems such as time-consuming, laborious, and low accuracy. However, the development of co-segregation molecular markers for...

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
  • Co-dominance InDel molecular marker for identifying single embryo and multiple embryos of citrus and application thereof
  • Co-dominance InDel molecular marker for identifying single embryo and multiple embryos of citrus and application thereof
  • Co-dominance InDel molecular marker for identifying single embryo and multiple embryos of citrus and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0052] Example 1 Identification of single polyembryos of 115 natural population materials such as tangerine, pomelo, and orange

[0053] 1. Preparation of genomic DNA of tangerine, grapefruit, orange, citrus, lemon and other citrus

[0054] In this example, the improved CTAB method was used to extract the genomic DNA of tangerines, grapefruits, oranges, citrus, lemons and other citrus; 2. Using primers to CKF / R, INSF / R and DelF / R amplification analysis of all the above Citrus genomic DNA;

[0055] The above primer pairs are:

[0056] Primer pair CKF / R

[0057] Forward primer 5'-CGTTCTTCAGACCCTTTCAG-3',

[0058] Reverse primer 5'-TGTCCTTTGTCCCATCCTTC-3';

[0059] Primer pair INSF / R

[0060] Forward primer 5'-GTATCCTAAAAATATAATCTTTATG-3',

[0061] Reverse primer 5'-ATCAATTTGTCCTTTGTCCC-3';

[0062] Primer pair DELF / R

[0063] Forward primer 5'-CAAAGATTATTATTGATATGGGAAAG-3',

[0064] Reverse primer 5'-CCTCGATGTACCTAACGATT-3'.

[0065] The PCR reaction system is as follows: 7μl PCRMix (purchased f...

Embodiment 2

[0067] Example 2 Identification of single polyembryos of the hybrid offspring of 100 HB pomelo and Fairchild tangerines

[0068] 1. Preparation of genomic DNA of 100 hybrids of HB pomelo and Fairchild tangerine

[0069] In this example, the improved CTAB method was used to extract the genomic DNA of the hybrid progeny of HB pomelo and Fairchild tangerine 100 strains.

[0070] 2. Using the above primer pairs CKF / R, INSF / R and DelF / R to amplify and analyze the genomic DNA of all the above citrus.

[0071] The PCR reaction system is as follows: 7μl PCRMix (purchased from Hubei Jingmao Company), 100ngDNA, forward and reverse primers (synthesized in Shenggong) each 0.2μM, ddH 2 O was added to the final volume of 15ul. The thermal cycling parameters are: 95°C for 5min; 95°C for 30sec, 58°C for 30sec, 72°C for 45sec, 35 cycles; 72°C for 10min, 1 cycle; 4°C storage, the reaction is completed on the S1000ThermalCycler PCR instrument. The amplified products were detected by 1.0% agarose gel el...

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 belongs to the field of citrus genomics and discloses a co-dominance InDel molecular marker for identifying single embryo and multiple embryos of citrus and application thereof. By carrying out depth re-sequencing on whole genome DNA of 107 materials including tangelos and oranges, whole genome sequence information is obtained, by means of comparing and screening, the gene sequence information in single-embryo and multi-embryo candidate areas is obtained, the sequence with the highest relevance degree to phenotype is screened out, and the sequence is an InDel sequence of 6bp. Accordingly, a set of simple, fast and co-dominant PCR markers are developed, and primers are used for carrying out PCR amplification on CK F/R, INS F/R and DEL F/R, so that the co-dominance InDel molecular marker for identifying the single embryo and the multiple embryos of the citrus is obtained. Finally, the marker is utilized for testing loci in 115 single-plant natural groups and 100 single-plant hybridized groups. The marker has the biggest advantages that cost is low, accuracy is high, and embryonic early selection can be achieved during the citrus seedling stage.

Description

Technical field [0001] The present invention relates to the field of citrus genomics, in particular to a co-dominant InDel molecular marker for identifying single embryos and polyembryos of citrus and its application. The molecular marker can be used as a marker for assisted selection of citrus single / multi-embryo breeding and is used to identify citrus single embryos. / Multi-embryonic varieties provide new molecular markers. Background technique [0002] Multi-embryo phenomenon is very common in citrus plants. One seed of multi-embryonic citrus can produce 2-10 embryos, and it is reported that a single seed has 42 embryos at most. Polyembryo is a special phenomenon of apomixis. It is obviously different from sexual reproduction (single embryo). It does not undergo the process of meiosis and fertilization, and develops into embryos from somatic cells (nucellus cells). The multi-embryonic process is asexual reproduction, and the offspring are actually clones of the mother (Clonal...

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): C12Q1/68C12N15/11C12N15/10
CPCC12Q1/6895C12Q2600/13C12Q2600/156
Inventor 徐强王霞徐远涛邓秀新
Owner HUAZHONG AGRI UNIV
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Try Eureka
PatSnap group products

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

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap