Inspection of wheat polyphenol oxidase activity characteristic and its special primer

A technology of phenol oxidase activity and wheat, which is applied in the field of detection of wheat polyphenol oxidase activity traits, can solve the problems such as undeveloped PPO active molecular markers, and achieve the effect of simple and easy detection method

Inactive Publication Date: 2007-09-05
CROP BREEDING & CULTIVATING INST CHINESE ACAD OF AGRI SCI
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Problems solved by technology

However, no molecular markers for PPO activity that can

Method used

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  • Inspection of wheat polyphenol oxidase activity characteristic and its special primer
  • Inspection of wheat polyphenol oxidase activity characteristic and its special primer

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0017] Example 1. Obtaining the SSR molecular marker Xgwm312 of wheat polyphenol oxidase gene PPO

[0018] Crossing the wheat variety'Zhongyou 9507' with high PPO activity cultivated by the Chinese Academy of Agricultural Sciences and the wheat variety "CA9632" with low PPO activity, F 1 Substitution was planted in the experimental field, and the anthers were cultured in vitro when heading, and haploid was doubled to obtain the DH population, and then the PPO activity analysis was performed.

[0019] Field trials were carried out in 3 locations, the Institute of Crop Sciences, Chinese Academy of Agricultural Sciences (Beijing), Cotton Institute of Chinese Academy of Agricultural Sciences (Anyang), and Shandong Academy of Agricultural Sciences (Jinan). Each location was repeated twice, and the community was divided into 3 rows. , The row length is 2 meters, and 80 seeds are sown in each row. The PPO activity of each DH line was tested after harvest.

[0020] With 126 pairs of SSR p...

Embodiment 2

[0022] Example 2. Determination of wheat polyphenol oxidase (PPO) activity

[0023] Experimental wheat varieties: 1. Suyin No. 10 (purchased from Crop Institute of Jiangsu Academy of Agricultural Sciences), 2. Nongda 152 (purchased from China Agricultural University), 3. Yumai 21 (purchased from Luohe Agricultural Science Research Institute, Henan Province) ), 4. Zhongliang 88375 (purchased from Tianshui Agricultural Science Institute of Gansu), 5. Yangmai No. 5 (purchased from Jiangsu Lixiahe Agricultural Science Institute), 6. Yumai 56 (purchased from Henan Academy of Agricultural Sciences wheat Institute), 7. Dongfeng 9801 (purchased from Institute of Crops, Chinese Academy of Agricultural Sciences), 8. Jinmai 50 (purchased from Cotton Research Institute of Shanxi Academy of Agricultural Sciences), 9. Gaocheng 8901 (purchased from Gaocheng Agriculture, Hebei Province) Institute of Science), 10. Gaoyou 503 (purchased from Shijiazhuang Institute of Agricultural Modernization, Chi...

Embodiment 3

[0031] Example 3. Validation verification of Xgwm312 molecular marker for detecting wheat polyphenol oxidase activity traits

[0032] Wheat varieties used in the experiment: the same as in Example 2.

[0033] The above 15 winter wheat varieties were planted in Cotton Research Institute of Chinese Academy of Agricultural Sciences (Anyang), Chinese Academy of Agricultural Sciences (Beijing) and Shandong Academy of Agricultural Sciences (Jinan). Field management is carried out as usual, and the seeds are extracted after harvesting. The genomic DNA of each wheat variety is then used as a template, and the nucleotide sequence of sequence 1 and the nucleotide sequence of sequence 2 in the sequence listing are used as primers for PCR amplification.

[0034] Among them, the PCR amplification reaction system is: 20μl total volume contains 1×PCR buffer (50mmol·L -1 KCl, 10mmol·L -1 Tris·Cl, 0.01% gelatin), MgCl 2 1.5mM, 250μM each of dNTP (A.T.C.G), 1U TaqDNA polymerase, 4pmol per primer, a...

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Abstract

The invention presents a measuring method of the activation of wheat polyphenol oxidase and its special primer. The primer provided by this invention, which is used to measure the activation of wheat polyphenol oxidase, is actually a pair of v formed by the nucleotide sequence of the sequence 1 and nucleotide sequence of the sequence 2 in the sequence list. The way of measuring the activation of wheat polyphenol oxidase is to make PCR amplification by taking the genome DNA of wheat waiting for measuring as the templet and the nucleotide sequence of the sequence 1 and nucleotide sequence of the sequence 2 in the sequence list as primer, and then test whether there is a stripe with the size of 198bp in the amplification amplified products. The method of this invention and its special primer will have an important role in the measuring of the activation of wheat polyphenol oxidase as well as in breeding.

Description

Technical field [0001] The invention relates to a method for detecting wheat polyphenol oxidase activity and its special primers. Background technique [0002] Wheat polyphenol oxidase (PPO) is the main factor that causes the color browning of wheat products such as noodles, steamed bread, dumplings and dough during storage, and it determines 70% of the color stability of noodles and other products. Reducing wheat PPO activity through genetic breeding and cultivating wheat varieties with low PPO activity is an important goal for wheat quality improvement. [0003] Researchers have done many genetic studies on the PPO activity of wheat. For example, Ge Xiuxiu and others have used a mixed genetic model of plant quantitative traits and multiple genes to analyze the PPO activity of winter wheat. The analysis results show that PPO activity is affected by two independent pairs. Genetic control. Jimenz and Anderson et al. also inferred that there are 1-2 major genes that control PPO in ...

Claims

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Application Information

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IPC IPC(8): C12Q1/68
Inventor 何中虎张立平孙道杰夏先春
Owner CROP BREEDING & CULTIVATING INST CHINESE ACAD OF AGRI SCI
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