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Tetraploid elytrigia elongata 3E chromosome molecular marker and application thereof

A technology of molecular marker, ryegrass, applied in the field of crop genetics and breeding

Active Publication Date: 2019-08-09
SICHUAN AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The present invention uses GBS sequencing to develop the molecular markers of tetraploid E. elongatum 3E chromosome, which has not been reported so far, which lays the foundation for further research and utilization of tetraploid E. elongatum in wheat genetics and breeding

Method used

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  • Tetraploid elytrigia elongata 3E chromosome molecular marker and application thereof
  • Tetraploid elytrigia elongata 3E chromosome molecular marker and application thereof
  • Tetraploid elytrigia elongata 3E chromosome molecular marker and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0058] Example 1 Based on GBS technology to obtain tetraploid E. elongatum 3E chromosome-specific fragments

[0059] When the materials such as diploid E. elongatum, tetraploid E. elongatum, parent 8801 and common wheat-tetraploid E. elongatum 1-7E chromosome substitution line grow to the seedling stage, take the leaves and send them to Beijing Novogene Technology Co., Ltd. carried out GBS sequencing, and obtained the effective sequence of each sample through GBS sequencing.

[0060] The sequence of the obtained wheat-tetraploid E. elongatum 3E / 3D substitution line was first compared with the known sequence of Chinese spring, and the sequence with a similarity of 22.85% (23%) or more to wheat was eliminated, and the remaining The following sequence was then compared with the sequence of 8801 and tetraploid E. elongatum to obtain a sequence with a similarity greater than 22.85% (23%). The remaining sequences were then compared with diploid E. elongatum and other substitution li...

Embodiment 2 4

[0061] Example 2 Development of Chromosome-Specific Molecular Markers of Tetraploid E. elongatum 3E

[0062] Primers were designed for the 269 specific sequences obtained above. The PCR primers were designed using the online software Primer3 Plus, and all the primers were synthesized at Chengdu Qingke Weiye Biotechnology Co., Ltd.

[0063]The PCR reaction system is as follows: the total volume of the PCR reaction is 25uL, DNA template (concentration needs to be diluted to about 100ng / uL) plus 1uL, upper and lower primers 1uL (10uM), 2×TaqMasterMix (P114) 12.5uL, ddH 2 O9.5uL.

[0064] The PCR program was: 94°C for 5h; 94°C for 30min, 50-60°C for 30min, 72°C for 2h, 35 cycles; 72°C for 10h; 12°C.

Embodiment 3

[0065] Embodiment 3 agarose gel electrophoresis detection

[0066] The PCR products were detected by 3% agarose gel electrophoresis, and the designed primers could amplify specific bands in tetraploid Echinopsis elongatum, parent 8801, and 3E / 3D substitution lines, and in CS, Shumai 482, Chuannong 16, diploid E. elongatum, 1E / 1D substitution line, 2E / 2A substitution line, 4E / 4D substitution line, 5E / 5D substitution line, 6E / 6D substitution line, 7E / 7D substitution line No bands could be amplified in the substitution lines, and this primer was the specific primer for the 3E / 3D substitution line, that is, the specific molecular marker for the 3E chromosome of tetraploid E. elongatum. 246 pairs of primers were successfully designed, and 146 pairs of specific primers were obtained after PCR amplification, product recovery, and sequencing verification in the above materials, and the molecular marker development efficiency reached 59.35%. The primer sequences of 20 pairs of specifi...

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Abstract

The invention discloses a tetraploid elytrigia elongata 3E chromosome molecular marker and application thereof. According to the molecular marker, with tetraploid elytrigia elongata genome DNA as a template, one of 1-20 primer pairs is adopted for amplification, and the molecular marker is obtained. By sequencing 1E-7E chromosome substitution lines of diploid and tetraploid elytrigia elongata, durum wheat-tetraploid elytrigia elongata amphidiploid and wheat-tetraploid elytrigia elongata, a specific sequence of a tetraploid elytrigia elongata 3E chromosome is obtained, the specific molecular marker of the 3E chromosome is developed, and verification is conducted for wheat and related genera species of the wheat. The molecular marker can be applied to quick detection of the tetraploid elytrigia elongata 3E chromosome and can accurately identify the chromosomes of the tetraploid elytrigia elongata, the diploid elytrigia elongata, the decaploid elytrigia elongata and other related genera species, and an important way is provided for phyletic evolution of triticeae, germplasm resource identification and auxiliary selection breeding of the molecular marker.

Description

technical field [0001] The invention belongs to the technical field of crop genetics and breeding, and specifically relates to a tetraploid Etyophony elongatum 3E chromosome molecular marker and an application thereof. Background technique [0002] Wheat (Triticum aestivum L.) is the food crop with the largest planting area in the world and its total output is second only to corn (Ge Jiangyan, 2012). It plays a pivotal role in human development. However, in recent years, with the change of farming system and the repeated use of a small number of parents in breeding (Zhang Lulu, 2016), the genetic diversity of wheat has decreased. There are abundant good genes in close relatives of wheat. At present, the good genes are mainly introduced into common wheat through distant hybridization, so as to enrich the genetic diversity of wheat. [0003] Elytrigia elongata (Host) Nevski is a close relative of wheat and belongs to the genus Elytrigia elongata (Host) Nevski. It is currently...

Claims

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Application Information

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IPC IPC(8): C12Q1/6895C12N15/11
CPCC12Q1/6895C12Q2600/13C12Q2600/156
Inventor 康厚扬龚壁燃谭彬文周永红徐黎黎王益曾建凡星沙莉娜张海琴
Owner SICHUAN AGRI UNIV
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