Detection primer, detection kit and detection method of dehiscence resistance of Sesamum indicum

A detection method and a technology for detecting primers, which are applied in the field of molecular biology, can solve the problems of cumbersome detection and low repetition rate of sesame, and achieve the effects of improving breeding efficiency, accurate detection methods, and shortening the breeding cycle

Active Publication Date: 2017-08-25
HENAN SESAME RES CENT HENAN ACADEMY OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] However, the above-mentioned traditional molecular markers have many defects such as cumbersome detection and low repetition rate when detecting the anti-capsulation resistance of sesame.

Method used

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  • Detection primer, detection kit and detection method of dehiscence resistance of Sesamum indicum
  • Detection primer, detection kit and detection method of dehiscence resistance of Sesamum indicum
  • Detection primer, detection kit and detection method of dehiscence resistance of Sesamum indicum

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0035] Acquisition of primers for detection of sesame resistance to dehiscence:

[0036] 1. Obtaining Zhengzhi InD01: using the anti-capsulation material "UCR82-15NS" (provided by D.M.Yermanos, Riverside, USA in 1983) as the female parent and Yuzhi No. 11 as the male parent, F 1 , selfing to obtain F 2 ; select F 2 A single plant of Zhongkang Xijiao was backcrossed with Yuzhi No. 11 to obtain BC 1 f 1 , selfing to get BC 1 f 2 ; select BC 1 f 2 A single plant of Zhongkang Xijiao was backcrossed with Yuzhi No. 11 to obtain BC 2 f 1 , selfing to get BC 2 f 2 ; select BC 2 f 2 A single plant of Zhongkang Xijiao was backcrossed with Yuzhi No. 11 to obtain BC 3 f1 , selfing to get BC 3 f 2 ; 2 generations of self-crossing, get BC 3 f 4 , the material with good capsule resistance, good capsule formation and disease resistance was selected and named Zhengzhi InD01.

[0037] Zhengzhi InD01, a sesame anti-capsulation-resistant material, Yuzhi No. 11, and its F2 segreg...

Embodiment 2

[0052] Sesame anti-capsulation detection kit, including: 10 μmol / μL 2 mL of the forward primer of Example 1, 10 μmol / μL of 2 mL of the reverse primer of Example 1, 1 mL of 5 U / μL Taq enzyme, 10 mL of 10×PCR buffer, 10 mM 2mL of dNTPs / μL, ddH 2 O 10mL.

Embodiment 3

[0054] Sesame anti-cracking capsule detection method, comprises the steps:

[0055] 1) Using the CTAB method to extract F from Zhengzhi InD01 and other sesame materials 2 The DNA of the leaf tissue of the progeny plants of the above-generation groups;

[0056] 2) PCR reaction system: 10 μL, containing 1 μL template DNA (25-50ng / μL), 0.1 μL Taq enzyme (5U / μL), 1 μL 10×PCR buffer, 0.2 μL dNTPs (10mM / μL), 0.2 μL of forward primer (10 μM / μL), 0.2 μL of reverse primer (10 μM / μL), 7.3 μL of ddH 2 O; Amplification program: pre-denaturation at 94°C for 1min; denaturation at 94°C for 30s, annealing at 57°C for 30s, extension at 72°C for 30s, 35 cycles, extension at 72°C for 10min;

[0057] 3) The amplified product was separated by 9% non-denaturing polyacrylamide gel electrophoresis, the electrophoresis buffer was 0.5×TBE, and the electrophoresis was separated at 150V constant power for 1h10min. After electrophoresis, the color was developed by silver staining, and the band pattern w...

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Abstract

The invention relates to a detection primer, detection kit and detection method of dehiscence resistance of Sesamum indicum, and belongs to the technical field of molecular biology. Compared with the sequence of a dehiscent Sesamum indicum gene, in the KANADI gene of anti-dehiscent Sesamum indicum, 14 bases are deleted in the DNA sequence, 77 bases are deleted in the cDNA sequence, the deleted bases are located between the 1,118th bp and the 1,332th bp in the downstream position of a transcription initiation codon, and the 14 bp bases are located in the connection position of a second exon and a second intron. According to the feature of the gene sequence differences, a detection primer of dehiscence resistance is designed, an important detection primer is provided for molecular marker assisted selection breeding of Sesamum indicum, and a rapid and accurate detection method is provided for screening of a new variety of Sesamum indicum with dehiscence resistance.

Description

technical field [0001] The invention relates to a detection primer, a detection kit and a detection method of sesame anti-capsulation resistance, and belongs to the technical field of molecular biology. Background technique [0002] Sesamum (Sesamum indicum L.) belongs to the family Flaxaceae, and is an important high-quality oil crop and characteristic agricultural product in my country, and it is also an important agricultural product for exporting foreign exchange. In recent years, the contradiction between the supply and demand of edible oil in our country is very prominent. Taking various measures to continuously increase the output of sesame and improve the production efficiency of sesame is a major topic of sesame research. However, the methods of sesame cultivation are backward, and manual operations are mainly used from sowing to harvesting. The original cultivation method leads to excessive labor input in sesame production, high labor intensity and low production e...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68C12N15/11
CPCC12Q1/6895C12Q2600/13C12Q2600/156
Inventor 刘艳阳郑永战梅鸿献武轲杜振伟崔承齐李小娟江晓琳
Owner HENAN SESAME RES CENT HENAN ACADEMY OF AGRI SCI
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