Primer set and method for identifying variety authentication and seed purity of tomatoes

An authentic tomato technology, applied in biochemical equipment and methods, microbiological determination/inspection, DNA/RNA fragments, etc., can solve the problems of low polymorphism, small tomato genetic variation, and few enzyme types

Active Publication Date: 2017-01-25
北京通州国际种业科技有限公司 +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The isoenzymes used for the purity identification of tomato varieties mainly include POD, EST, ACP, ADH, MDH, etc. However, due to the small genetic variation and low polymorphism of cultivated tomato, there are few available enzymes, and the e

Method used

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  • Primer set and method for identifying variety authentication and seed purity of tomatoes
  • Primer set and method for identifying variety authentication and seed purity of tomatoes
  • Primer set and method for identifying variety authentication and seed purity of tomatoes

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0079] Embodiment 1, identification or auxiliary identification tomato variety authenticity and the preparation of seed purity of a set of primers

[0080] The set of primers provided by the present invention for identifying or assisting the identification of the authenticity and seed purity of tomato varieties is a set of primers 1 and a set of primers 2, and the names of the set of primers 1 are respectively Sol_SSR19, Sol_SSR120, Sol_SSR305, Sol_SSR18, Sol_SSR132, Sol_SSR3, Sol_SSR110, Sol_SSR146 、Sol_SSR125、Sol_SSR117、Sol_SSR101、Sol_SSR218、Sol_SSR225、Sol_SSR97、Sol_SSR185、Sol_SSR233、Sol_SSR184、Sol_SSR254、Sol_SSR318、Sol_SSR272、Sol_SSR68、Sol_SSR169、Sol_SSR13和Sol_SSR96的24对引物;成套引物2为名称分别为Sol_SSR190、Sol_SSR303、Sol_SSR302、 Sol_SSR148、Sol_SSR304、Sol_SSR45、Sol_SSR24、Sol_SSR76、Sol_SSR306、Sol_SSR308、Sol_SSR201、Sol_SSR23、Sol_SSR126、Sol_SSR312、Sol_SSR224、Sol_SSR313、Sol_SSR239、Sol_SSR315、Sol_SSR69、Sol_SSR266、Sol_SSR317、Sol_SSR283、Sol_SSR321和Sol_SSR300的24对引物。

[0081] The 5' end of the up...

Embodiment 2

[0087] Example 2. Using a set of primers to identify or assist in identifying the authenticity and seed purity of tomato varieties to identify the authenticity of tomato varieties

[0088] (1) The tomato material for constructing the map was provided by Mr. Li Changbao from the Vegetable Center of Beijing Academy of Agriculture and Forestry Sciences. From 402 materials, 100 varieties were randomly selected (Table 2). The selected hybrids are listed in Table 3.

[0089] Table 2, 100 test tomato varieties

[0090]

[0091]

[0092] Table 3. Materials of 12 representative tomato hybrid varieties

[0093] breed name category traits Remark Jing Fan 104 big powder pink oversized fruit hybrid Beijing Fan 203 big powder medium to large pink fruit hybrid Beijing Fan 501 big red red medium to large fruit hybrid Beijing Fan 601 big red large red fruit hybrid Jing Fan 701 rhubarb large yellow fruit hybrid J...

Embodiment 3

[0126] Example 3: Identification of the purity of tomato hybrid seeds by using a set of primers for identification or auxiliary identification of tomato variety authenticity and seed purity

[0127] The experiment was repeated three times, and the specific steps of each repeated experiment were as follows:

[0128] (1) sample

[0129] Randomly select 100-200 tomato seeds from the seeds of the submitted tomato variety JF101 (Beijing Academy of Agriculture and Forestry Sciences) for cultivation, and obtain no less than 94 single tomato plants, together with the male and female parents of the variety Y15-013 \Y15-049 (male and female as control samples, Beijing Academy of Agriculture and Forestry Sciences) plants, a total of 96 samples were obtained.

[0130] (2) Extracting the DNA of the tested tomato variety: the DNA of the above-mentioned 96 samples was extracted by the CTAB method, and the specific extraction method was the same as step (2) of Example 2.

[0131] (3) PCR am...

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Abstract

The invention discloses a primer set and method for identifying variety authentication and seed purity of tomatoes. Sequences of 48 pairs of primers of the primer set for identifying variety authentication of tomatoes and purity of seeds is successively as shown in sequences 1-96 in a sequence table, a sequence 2n and a sequence 2n-1 form a primer pair, and n is a natural number in 1-24. The method for identifying variety authentication and seed purity of tomatoes comprises the following steps: carrying out PCR amplification on DNA of tomatoes to be detected by using the primer set; and determining variety authentication and seed purity of the tomatoes to be detected according to difference of PCR products of the tomatoes to be detected. Experiments prove that the primer set not only can be used for identifying the variety authentication of the tomatoes, but also can be used for identifying the seed purity of the tomatoes, and has the advantages that the accuracy is high, results are stable, the primer set is not affected by environment conditions and development stages, statistics is simple and convenient and the like.

Description

technical field [0001] The invention relates to a set of primers and a method for identifying tomato variety authenticity and seed purity in the field of biotechnology. Background technique [0002] Tomato (Lycopersicon esculentum L.) is one of the most important vegetable crops in the world with high nutritional value. Due to the unique edible, health care and auxiliary therapeutic effects of tomato and its products, it occupies an important position in the world's agricultural production and trade. According to statistics released by the World Processed Tomato Council (WPTC): In 2013, the global output of processed tomatoes was 33.012 million tons. Along with the advantageous development of my country's tomato seed industry, some decked varieties appeared on the market, shoddy, affecting the output and income of growers; even people illegally stole breeding materials for breeding, which seriously affected my country's breeding units Therefore, the rapid and accurate ident...

Claims

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Application Information

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IPC IPC(8): C12Q1/68C12N15/11
CPCC12Q1/6895C12Q2600/156
Inventor 刘宝平王喜萍李常保王铭堂张东峰马俊召王亚楠周明刘海燕杜敏敏
Owner 北京通州国际种业科技有限公司
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