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A special primer, kit and application thereof for detecting the presence or absence of yellow leaf gene in wheat

A kit and yellow leaf technology, applied in the field of molecular genetics, can solve problems such as unreported research on common wheat yellow leaf genes, and achieve the effects of improving breeding efficiency, low cost and simple operation.

Inactive Publication Date: 2021-08-27
HENAN INST OF SCI & TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, there is no report on the gene controlling the yellow leaves of common wheat

Method used

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  • A special primer, kit and application thereof for detecting the presence or absence of yellow leaf gene in wheat
  • A special primer, kit and application thereof for detecting the presence or absence of yellow leaf gene in wheat
  • A special primer, kit and application thereof for detecting the presence or absence of yellow leaf gene in wheat

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0035] Design and screening process of primers used in the present invention.

[0036] (1) Acquisition of yellow leaf wheat material and its main agronomic characters

[0037] A naturally mutated yellow leaf mutant Yglw-1 was obtained from the hybrid progeny of "Bainong 58409 / Fortune Mai". At the jointing stage of wheat, the bottom leaves began to turn yellow, and the yellow leaves gradually spread to the upper part of the plant until maturity. Compared to its sister line Cf5019-21 (normal leaf color), Yglw-1 had significantly reduced ear length, grain number per ear, and thousand-grain weight (see Table 1).

[0038] Table 1 Comparison of main agronomic characters of yellow leaf mutant Yglw-1 and its sister line Cf5019-21

[0039]

[0040] *Indicates significant difference at the 0.05 level

[0041] (2) Genetic analysis of yellow leaf mutant Yglw-1

[0042] F was obtained by crossing the yellow leaf mutant Yglw-1 with its sister line Cf5019-21 1 seeds, F 1 Self-cross ...

Embodiment 2

[0055] The nucleotide sequence of the primer for detecting the yellow leaf gene of wheat in the present embodiment is:

[0056] F-1: 5'-TGACGTGACGAAAAGTGGAG-3', R-1: 5'-CTTCAATGATTCCCGGAAGA-3'.

[0057] The kit in this example includes the above primers, and also includes 10 × Buffer (Mg 2+ ), dNTPs and Taq DNA polymerase.

[0058] Use the above-mentioned primers to detect the yellow leaf gene of wheat, specifically including:

[0059] 1) Extract (Yglw-1×Cf5019-21)F in Example 1 5 Genomic DNA of some yellow leaf plants and normal plants in the population;

[0060] 2) Using the DNA of the sample to be tested as a template, use the above primers for PCR amplification. The PCR reaction system is 20 ul, containing 10 × Buffer (Mg 2+ ) 2.0μL, 2.5mmol / L dNTPs 0.6μL, 5U / μL Taq DNA polymerase 0.2μL, 10μmol / L primer 2.0μL, 30ng / μL DNA template 2.0μL, ddH 2 O 13.2 μL.

[0061] The PCR amplification program was as follows: pre-denaturation at 94°C for 5 min; denaturation at 94°C for ...

Embodiment 3

[0064] The nucleotide sequence of the primer for detecting the yellow leaf gene of wheat in the present embodiment is:

[0065] F-2: 5'-TTTTGGTGTTCTTTTTGGGG-3', R-2: 5'-CCAGCTTCTTCTCGCCTCTA-3'.

[0066] The kit in this example includes the above primers, and also includes 10 × Buffer (Mg 2+ ), dNTPs and Taq DNA polymerase.

[0067] Use the above-mentioned primers to detect the yellow leaf gene of wheat, specifically including:

[0068] The DNA of the sample to be tested in Example 2 was used as a template, and the above primers were used for PCR amplification. The PCR reaction system was 20 ul, containing 10 × Buffer (Mg 2+ ) 2.0μL, 2.5mmol / L dNTPs 0.6μL, 5U / μL Taq DNA polymerase 0.2μL, 10μmol / L primer 2.0μL, 30ng / μL DNA template 2.0μL, ddH 2 O 13.2 μL.

[0069] The PCR amplification program was as follows: pre-denaturation at 94°C for 5 min; denaturation at 94°C for 1 min, annealing at 60°C for 1 min, extension at 72°C for 2 min, 35 cycles; extension at 72°C for 5 min. T...

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Abstract

The invention relates to a special primer for detecting whether there is a yellow leaf gene in wheat, a kit and an application thereof, belonging to the technical field of molecular genetics. The primers for detecting whether there is a yellow leaf gene in wheat include BE494262, CD453721 and / or BF202681. Use primers BE494262, CD453721 and / or BF202681 to perform PCR amplification on the wheat genomic DNA respectively. If the PCR amplification obtained by these 3 primers If there are bands of 420bp, 1500bp and 580bp in the product, the yellow leaf gene is present in the wheat to be tested. The three special primers BE494262, BF202681 and / or CD453721 of the present invention can accurately detect whether the wheat yellow leaf gene exists, can be used for early generation selection of breeding, shorten the breeding cycle, improve breeding efficiency, and have the advantages of simple operation and low cost.

Description

technical field [0001] The invention relates to a special primer, a kit and application for detecting whether there is a yellow leaf gene in wheat, and belongs to the technical field of molecular genetics. Background technique [0002] Leaves are the main organ for photosynthesis in plants. Leaf regulates plant growth and development by converting light energy into chemical energy through photosynthesis. Leaf color mutations often directly or indirectly affect the biosynthesis or degradation of chlorophyll, thereby affecting the content of chlorophyll, so it is also called chlorophyll deletion mutation. Leaf color variation usually reduces the photosynthesis of mutants, resulting in reduced crop yields and even plant death in severe cases, so it was considered a meaningless mutation for a long time. However, leaf color mutants have received a lot of attention from geneticists and molecular biologists in recent years. Since abnormal leaf color change affects the function a...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/6895C12N15/11
CPCC12Q1/6895C12Q2600/156
Inventor 李小军茹振钢姜小苓李淦陈向东丁位华胡喜贵王玉泉吴晓军
Owner HENAN INST OF SCI & TECH
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