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A molecular marker and its application for rapid detection of leaf rust resistance gene in Thiopyrum elongatum

A technology of leaf rust resistance gene and Echinopsis elongatum, which is applied in the field of molecular genetic breeding, can solve the problems of common wheat chromosome exchange, difficulty in the application of genetic breeding, difficult fine positioning and closely linked marker development, etc., to achieve convenient cloning and improve Breeding efficiency, using accurate results

Active Publication Date: 2021-08-24
SHANDONG AGRICULTURAL UNIVERSITY
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  • Abstract
  • Description
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AI Technical Summary

Problems solved by technology

However, because it is derived from the chromosome of E. elongatum, it is difficult to exchange with common wheat chromosomes, which makes it difficult to fine-map and develop closely linked markers, which brings great difficulties to the application in genetic breeding.

Method used

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  • A molecular marker and its application for rapid detection of leaf rust resistance gene in Thiopyrum elongatum
  • A molecular marker and its application for rapid detection of leaf rust resistance gene in Thiopyrum elongatum

Examples

Experimental program
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Effect test

Embodiment 1

[0033] Example 1 Creation of short-segment materials resistant to leaf rust of wheat-Thinopyrum elongatum

[0034] In 2015, the hybrid combination (KS11695×CS Ph1b mutant) was configured in the greenhouse of Shandong Agricultural University and obtained F 1 ; In 2016, will receive F 1 with CS- Ph1b Mutants are hybridized to obtain BC 1 f 1 , and carry out molecular marker identification on the obtained plants, on the basis of molecular marker identification, the BC carrying target gene and exogenous fragment 1 f 1 BC 1 f 2 . In 2016 and 2017, with the help of molecular markers, short-segment translocation lines of wheat-Thinopyrum elongatum were obtained, and their resistance to leaf rust was identified.

Embodiment 2

[0035] Example 2 Anti-rust gene of Thiopyrum elongatum Lr19 Development of Tightly Linked Molecular Markers

[0036] According to the published sequence information of the D genome of A. taustus, and downloading the scaffold near the end of the 7DL chromosome, the Chinese spring mutant, K11695, 7el 1 (7D) and Thinopyrum ponticum were used as materials for PCR amplification to search for molecular markers closely linked to the leaf rust resistance gene Lr19.

[0037] In the PCR amplification reaction system, the PCR reagent composition is: 2 μl template containing 50-100ng DNA (from Chinese spring mutant, K11695, 7el 1 (7D) and Genomic DNA extracted from Th. ponticum), 1.5μl 10× PCR buffer (containing Mg 2 + ), 1.2 μl ldNTP, left and right primers (SEQ ID No.1 and SEQ ID No.2), 0.15 μl R-Taq DNA polymerase, 8.15 μl H 2 O. The PCR amplification program was pre-denaturation at 94°C for 5 min; then denaturation at 94°C for 50 s, annealing at 49°C for 40 s, and extension at 72...

Embodiment 3

[0038] Example 3 Leaf Rust Resistance Gene of Echinopsis elongatum Lr19 Validation of tightly linked molecular markers

[0039] to 7el 1 、7el 2 , Thinopyrum ponticum, k11695, CS- ph1b , JM22, LDN, 2147, and short-fragment screening offspring were used as experimental materials to verify the obtained molecular marker Xsdau798,

[0040] In the PCR amplification reaction system, the PCR reagent composition is: 2 μl template containing 50-100ng DNA (from 7el 1 、7el 2 , Thinopyrum ponticum, k11695, CS- ph1b , JM22, LDN, 2147, genomic DNA extracted from short fragment screening offspring), 1.5μl 10× PCR buffer (containing Mg 2+ ), 1.2 μl dNTP, left and right primers (SEQ ID No.1 and SEQ ID No.2, SEQ ID No.3 and SEQ ID No.4), 0.15 μl R-Taq DNA polymerase, 8.15 μl H 2 O. The PCR amplification program was pre-denaturation at 94°C for 5 min; then denaturation at 94°C for 50 s, annealing at 49°C for 40 s, and extension at 72°C for 60 s, 35 cycles; finally, extension at 72°C for...

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Abstract

The present invention provides a molecular marker and its application for rapid detection of the leaf rust resistance gene of E. elongatum. With the aid of the induced short-segment translocation line of E. elongatum for resistance to leaf rust, combined with the reference sequence of the wheat D genome, The molecular marker Xsdau798 closely linked to it was designed and developed for wheat leaf rust resistance breeding and early molecular marker-assisted selection of leaf rust resistance traits to improve breeding efficiency.

Description

technical field [0001] The invention relates to the field of molecular genetic breeding, and specifically relates to a molecular marker for rapid detection of the leaf rust resistance gene of Echinopsis elongatum and its application. Background technique [0002] Wheat leaf rust is caused by the fungus Phytophthora tritici ( Puccinia recondita f .sp .tritici ) caused major fungal diseases, which bring large yield losses to wheat production worldwide every year. In recent years, with climate change, wheat leaf rust has begun to break out on a large scale. Therefore, the control of wheat leaf rust is an important issue in wheat production. Experience has proved that cultivating disease-resistant wheat varieties is the safest, economical, effective and environmentally friendly measure to control and reduce damage. However, the frequent mutation of the virulence genes of leaf rust and large-scale single planting of disease-resistant varieties often lead to the "loss" of resist...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/6895C12N15/11
Inventor 王宏伟孔令让吕忠璠马信
Owner SHANDONG AGRICULTURAL UNIVERSITY
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