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Wheat EST (expressed sequence tag) sequence, molecular marking for stripe rust-resistance property, and application

A technology for stripe rust and wheat, which is applied in the field of crop breeding and molecular genetics, can solve the problems of lower accuracy rate, influence of resistance gene breeding, inability to eliminate self-defects, etc., and achieve reliable detection, optimal annealing temperature and annealing effect

Inactive Publication Date: 2013-08-28
SICHUAN AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although this method can meet the needs of rapid identification, its accuracy and reliability are limited.
The technical defect is that this method only uses linked or closely linked SSR molecular markers, and since the SSR sequence itself is a simple repeat sequence, the probability of variation is much higher than other sequences
Once the SSR sequence mutates, it will be directly displayed on the amplified band, and the wheat that originally contained the resistance gene will be missed during the identification process, which will affect the reliability of identification.
Since the identification of stripe rust resistance genes by SSR markers has its own defects that cannot be eliminated, the use of closely linked molecular markers to identify stripe rust resistance genes cannot be accurate, which will have a certain impact on the breeding of resistance genes and affect the breeding process
In addition, many stripe rust resistance genes have not yet been finely mapped, and the accuracy of identification of these stripe rust resistance genes will be further reduced

Method used

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  • Wheat EST (expressed sequence tag) sequence, molecular marking for stripe rust-resistance property, and application
  • Wheat EST (expressed sequence tag) sequence, molecular marking for stripe rust-resistance property, and application
  • Wheat EST (expressed sequence tag) sequence, molecular marking for stripe rust-resistance property, and application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0055] This example describes a molecular marker method in which the target sequence contains the wheat EST sequence JK972238-94, which is based on PCR for EST markers.

[0056] 1. Unidentified wheat materials

[0057] Wheat stripe rust-resistant line L693 (it was identified that this material carries a dominant gene controlling stripe rust resistance) and susceptible line L661 were germinated indoors, and when the plants grew to the three-leaf stage, they were treated with stripe rust. The physiological race "Tiaozhong 32" was inoculated by smearing method to induce the expression of resistance gene ( figure 1 ).

[0058] 2. Extraction of total DNA from wheat materials to be identified

[0059] 24, 48, and 72 hours after inoculation, the leaves of disease-resistant and susceptible materials were taken and stored at -80°C, and the sampled leaves were ground to powder with liquid nitrogen, and the total DNA of wheat materials was extracted according to the CTAB method.

[00...

Embodiment 2

[0071] This example describes a method for identifying resistance to wheat stripe rust.

[0072] The amplified products obtained in Example 1 were compared with their corresponding standard target sequences, and if the amplified products were consistent with the length of the standard target sequence (see Table 1), the strain of wheat contained stripe rust resistance genes, and should show resistance to stripe rust. stripe rust traits; on the contrary, it means that the single wheat plant does not contain the stripe rust resistance gene and should be susceptible to stripe rust.

[0073] figure 2 shown. Identification of F in wheat material L693×L661 using co-isolated primer LSc18 2 The length of the amplified product of the wheat single plant containing the stripe rust resistance gene was 164bp, while that of other wheat single plants not containing the stripe rust resistance gene had no such amplification product. L693×L661F 2 Population expansion such as figure 2 , th...

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Abstract

The invention discloses an EST (expressed sequence tag) sequence related to wheat stripe rust-resistance property, a DNA (deoxyribonucleic acid) molecular marking method and an application for the EST sequence. The invention provides an EST sequence expressed by a wheat stripe rust-resistance gene, and a DNA molecular marking method for identifying wheat stripe rust-resistance property based on the EST sequence, aiming at the defects existing in identification for wheat stripe rust-resistance property by virtue of an SSR (simple sequence repeat) marking method in the prior art. The invention provides a wheat EST sequence JK972238-94, wherein the base sequence is shown in SEQ ID NO. 1, and the function annotation is the coded sequence of a heavy metal binding protein; and the five target sequences, namely LS36, LSc1617, LSc18, LSc1517 and LSc1718, in the EST marking method, and the specific primer pairs of the respective target sequences are provided; each specific primer pair is pleomorphic, capable of being subjected to co-segregation with resistance genes, and capable of being used for molecular marker assistance breeding; and control conditions of a PCR (polymerase chain reaction) process in the EST marking method are provided. The EST sequence disclosed by the invention is applied to identification for wheat stripe rust-resistance, and capable of accurately and reliably detecting wheat stripe rust resistance genes.

Description

technical field [0001] The invention relates to a wheat EST sequence and a wheat DNA molecular marker method, in particular to an EST sequence related to wheat stripe rust resistance traits and a DNA molecular marker method and application thereof, belonging to the fields of molecular genetics and crop breeding. Background technique [0002] Wheat stripe rust is a worldwide important disease that threatens wheat production. Stripe rust spreads through the air, and has a wide range of damage and a high prevalence rate. It is the key monitoring and control object in the wheat planting process. Wheat growing areas all over the world urgently need to find and introduce new disease resistance genes to enhance the resistance of wheat cultivars to meet production needs. Practice has proved that for wheat diseases, cultivating and promoting disease-resistant varieties is the most economical, effective and safe means of prevention and control. Discovering and quickly identifying str...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/11C12Q1/68
Inventor 罗培高李鑫黄强张怀渝张敏陈万权任正隆谭飞泉
Owner SICHUAN AGRI UNIV
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