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A long-spiked wheatgrass e e Genome-specific molecular markers and their applications

It is a technology for the development and application of tetraploid wheatgrass and genome, which can be applied in the directions of recombinant DNA technology, microbial determination/inspection, DNA/RNA fragment, etc.

Active Publication Date: 2022-07-12
SICHUAN AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The development of the above molecular markers mainly comes from diploid E. elongatum and decaploid E. elongatum, while tetraploid E. elongatum has fewer molecular markers, which hinders the development of tetraploid E. elongatum genes. Resource Development and Application

Method used

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  • A long-spiked wheatgrass e <sup>e</sup> Genome-specific molecular markers and their applications
  • A long-spiked wheatgrass e <sup>e</sup> Genome-specific molecular markers and their applications
  • A long-spiked wheatgrass e <sup>e</sup> Genome-specific molecular markers and their applications

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0034] Embodiment 1 obtains tetraploid R. longa 3E based on GBS technology e Chromosome specific segment

[0035] Diploid S. serratus, tetraploid S. sylvaticus, parent 8801 and common wheat-tetraploid S. sylvaticus 1-7E e When the chromosome substitution lines and other materials grew to the seedling stage, the leaves were taken and sent to Beijing Nuohezhiyuan Technology Co., Ltd. for GBS sequencing, and the effective sequences of each sample were obtained by GBS sequencing. The obtained wheat-tetraploid R. longa 3E e The sequences of the / 3D substitution line were firstly aligned with the known sequences of Chinese spring, and the sequences with a similarity of more than 22.85% (23%) to wheat were eliminated, and the remaining sequences were compared with 8801 and tetraploid long panicle. The sequences of wheatgrass were compared to obtain sequences with a similarity greater than 22.85% (23%), and the remaining sequences were compared with the sequences of the diploid whea...

Embodiment 2

[0036] Embodiment 2 E. long spike E e Development of genome-specific molecular markers

[0037] The primers were designed for the specific sequences obtained above. The online software Primer3Plus was used for PCR primer design. All primers were synthesized in Chengdu Qingke Weiye Biotechnology Co., Ltd.

[0038] The PCR system is as follows: the total volume of the PCR reaction is 25uL, the DNA template (the concentration needs to be diluted to about 100ng / uL) add 1uL, the upper and lower primers are each 1uL (10uM), 2×Taq Master Mix (P114) 12.5uL, ddH 2 O 9.5uL.

[0039] The PCR program was: 94°C for 5h; 94°C for 30min, 50-60°C for 30min, 72°C for 1h, 35 cycles; 72°C for 10h; 12°C.

Embodiment 3

[0040] Example 3 Detection by agarose gel electrophoresis

[0041] The PCR products were detected by 3% agarose gel electrophoresis, and the designed primers were used in diploid R. longa, tetraploid R. longa, parental 8801, 3E e / 3D substitution line can amplify specific bands, in CS, Shumai 482, Chuannong 16, 1E e / 1D substitution system, 2E e / 2A substitution system, 4E e / 4D substitution system, 5E e / 5D substitution system, 6E e / 6D substitution system, 7E e There is no band in the / 7D substitution line, and we think this primer may be Trichoderma ulmoides 3E e Chromosome-specific primers, i.e. R. longiflorum 3E e Chromosome-specific molecular markers. After PCR product recovery and sequencing verification, the amplification of the specific molecular marker Primer3E-219 in the above materials is shown in the appendix figure 1 Shown: only in diploid R. thorn, tetraploid R. thorn, 8801 and 3E e Bands can be amplified in the / 3D substitution line, but no bands were ...

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Abstract

The invention discloses a kind of wheatgrass E. e Genome-specific molecular markers and applications thereof, specifically for diploid R. longa, tetraploid R. longa, durum wheat-tetraploid D. longa diploid 8801, and common wheat-tetraploid Erythorax longus 1E e ‑7E e The chromosomal replacement line was sequenced, and the sequence alignment was performed to obtain a tetraploid Trichoderma 3E e Chromosome-specific sequence, R. longiflorum 3E was developed e Chromosome-specific molecular markers and validated in wheat and its relatives. The specific molecular marker developed in the present invention can not only be used for diploid, tetraploid, and decoploid Trichoderma splendens E e Rapid detection of genomes, also available for all Triticeae containing E e Chromosome or chromosome fragment identification of genomic species, and can accurately identify and distinguish E e and E b Genome, which provides the basis for the phylogeny, germplasm identification, and molecular marker-assisted selection breeding of Triticum species.

Description

technical field [0001] The invention belongs to the technical field of crop genetics and breeding, and in particular relates to a kind of E. e Genome-specific molecular markers and their applications. Background technique [0002] Wheat (Triticum aestivum L.) is the food crop with the largest planting area in the world and second only to corn in total output (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 few parents in breeding (Zhang Lulu, 2016), the genetic diversity of wheat has been reduced. There are abundant good genes in the relative species of wheat. At present, its 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 closely related species of wheat, belonging to the genus Elytrigia of the Triticaceae family, and it is currently recognize...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/6895C12N15/11
CPCC12Q1/6895C12Q2600/156C12Q2600/13
Inventor 康厚扬张桔维李三月习玲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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