Auxiliary identification method of wheat with high kernel lipoxidase activity character and special primers thereof
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A technology for lipoxygenase and oxidase activity, which is applied in biochemical equipment and methods, measurement/testing of microorganisms, DNA/RNA fragments, etc., and can solve problems such as undeveloped LOX gene molecular markers
Inactive Publication Date: 2010-08-25
INST OF CROP SCI CHINESE ACAD OF AGRI SCI
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The complete sequence of the LOX gene of common wheat has not been cloned, and the complete sequence of the LOX gene cDNA has been cloned, but no molecular markers of the LOX gene that can be applied to wheat breeding have been developed so far
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Embodiment 1
[0023] Example 1, the acquisition of the SSR molecular marker combination Xwmc312 / Xgwm251 of the wheat LOX gene
[0024]The wheat variety 'Zhongyou 9507' with high LOX activity bred by the Chinese Academy of Agricultural Sciences was crossed with the wheat variety 'CA9632' with low PPO activity, F 1 Subplanted in the experimental field, the anthers were cultured in vitro when heading, and the haploid was doubled to obtain the DH population, and then the LOX activity was analyzed.
[0025] The field experiments were carried out at two sites, the Cotton Research Institute of the Chinese Academy of Agricultural Sciences (Anyang) and the Crops Institute of the Shandong Academy of Agricultural Sciences (Jinan). Each site was repeated twice. grain. Each DH line was tested for LOX activity after harvest.
[0028] Embodiment 2, the mensuration of wheat lipoxygenase (LOX) activity
[0029] The used wheat variety of embodiment:
[0030] (1) Heng 7228, purchased from the Institute of Dry Farming, Hebei Academy of Agriculture and Forestry);
[0031] (2) Gaocheng 8901, purchased from Gaocheng Agricultural Science Research Institute of Hebei Province);
[0032] (3) Taishan 269, purchased from the Crop Institute of Shandong Academy of Agricultural Sciences;
[0033] (4) Zhou 892, purchased from Zhoukou Academy of Agricultural Sciences, Henan Province;
[0034] (5) Guan 35 was purchased from the Institute of Dry Farming, Hebei Academy of Agriculture and Forestry Sciences;
[0035] (6) Shixin 733, purchased from Shijiazhuang New Wheat Variety and New Technology Research Institute;
[0036] (7) Beijing 9428, purchased from Beijing Seed Company;
[0037] (8) CA0431; obtained from the National Germplasm Bank of the Institute of Crop Science, Chinese Academy of Agricultural Sciences;
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Abstract
The invention discloses an auxiliary identification method of wheat with a high kernel lipoxidase activity character and special primers thereof. The method comprises the following steps of: simultaneously carrying out PCR (Polymerase Chain Reaction) amplification by using a primer pair A and a primer pair B by taking gene group DNA of wheat to be tested as a template; and if DNA segments of 235 bp and 117bp are obtained, judging the wheat to be tested is candidate wheat with the high kernel lipoxidase activity character, wherein the high kernel lipoxidase activity character means the lipoxidase activity of a kernel is above 75 A234min-1g-1, the primer pair A refers to a primer pair consisting of DNA shown as sequences 1 and 2 in a sequence table, and the primer pair B refers to a primer pair consisting of DNA shown as sequences 3 and 4 in the sequence table. The method is simple, easy, stable and reliable and can carry out molecular marker assisted selection on wheat lipoxidase gene LOX to culture high-quality wheat breed with high lipoxidase activity and can test the lipoxidase activity in wheat. The method and the special primers are about to play a very important role in wheat lipoxidase activity character detection and wheat breading.
Description
technical field [0001] The invention relates to a method for assisting identification of wheat with the trait of high grain lipoxygenase activity and special primers thereof. Background technique [0002] The color of flour and its products is an important content of genetic improvement of wheat quality. The activity of lipoxygenase (LOX) is the main factor affecting the color and processing quality of flour products, and it causes the color loss and whitening of flour and flour products during the processing and storage of wheat products. Improving wheat LOX activity through genetic breeding and cultivating wheat varieties with high LOX activity is an important goal of wheat quality improvement. [0003] Researchers have done many genetic studies on wheat LOX activity. Borrelli et al. (1999) showed that genotype, environment, and gene-environment interaction have significant effects on LOX activity, but it is mainly affected by genetic factors. Leenhardt et al. (2006) stu...
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