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Method for quickly and accurately detecting purity of hybrid rice seed

A technology of hybrid rice and seeds, applied in the field of genetics, can solve the problems of unsure whether the seeds are qualified, plant traits are affected by the environment, and affect seed sales and marketing, etc., to meet the requirements of detection speed and accuracy, and to meet the timely market sales , the effect of heavy workload

Active Publication Date: 2014-07-23
海南广陵高科实业有限公司 +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The shortcomings of field identification are obvious: first, it needs to be planted for one season, which takes a long time, takes up a lot of land, and is expensive. However, the time from production to sale of seeds is only 2-3 months. The field identification period seriously affects the sale of seeds.
The second is that plant traits are often affected by the environment. For example, under high fertility conditions, the plants are taller and larger, so they are misjudged as miscellaneous plants, making the identification results inaccurate.
Therefore, even if the test results are obtained, it is not sure whether the batch of seeds is qualified, which brings huge risks to the market.

Method used

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  • Method for quickly and accurately detecting purity of hybrid rice seed
  • Method for quickly and accurately detecting purity of hybrid rice seed
  • Method for quickly and accurately detecting purity of hybrid rice seed

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0023] Seed purity detection method of hybrid rice variety "Keyou 73" (see figure 1 ):

[0024] (1) "Keyou 73" hybrid seed sampling

[0025] "Keyou 73" is a new hybrid rice variety selected and bred, and has applied for new variety protection. Its male parent is R7723 and its female parent is Jinke 1A. Sampling analysis of 127 batches of "Keyou 73" seed purity. Using the "binomial distribution overall mean confidence interval" function in the DPS (Data Processing System: Data Processing System.) 2000 software, it is calculated that the sampling sample needs to reach 3000 samples to meet the requirement that the detection error does not exceed 1%. Therefore, using a seed sampler, according to the principle of random sampling, approximately 4,500 seeds are drawn from each batch of seeds.

[0026] (2) Seed germination, sample mixing and DNA extraction

[0027] Spread 5 layers of newspapers on a 40cm×80cm porcelain plate, add a small amount of water until the newspapers are co...

Embodiment 2

[0035] The detection method of the present embodiment is basically the same as embodiment 1, the difference is:

[0036] The steps of DNA extraction in step (2) are:

[0037] When the seeds germinate and grow to the 7th to 10th day, 1 leaf has grown. At this time, select 3000 seedlings, and each seedling takes 1 leaf, according to the plant genome DNA extraction kit (DP305, Tiangeng, Beijing) operation manual Separation and purification of genomic DNA, extraction of DNA from each leaf, quantitative extraction of DNA, and equal mixing; ND2000 ultra-micro spectrophotometer was used to measure the concentration and purity of samples. A total of 127 pooled DNA samples were obtained, each representing 127 sampled samples.

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Abstract

The invention discloses a method for quickly and accurately detecting the purity of a hybrid rice seed, which comprises the following steps: according to seed purity requirements, calculating the quantity of samples; sampling in a quantity-equal manner and mixing extracted DNA (deoxyribonucleic acid), or respectively extracting DNA and then mixing in a quantity-equal manner; designing PCR (polymerase chain reaction) primers according to the base sequence of detected sites, and amplifying the detected sites; recovering and purifying the PCR amplification products; building a library by using the amplification products, and performing high-flux sequencing; and calculating the seed purity according to the sequence proportion of the detected sites. The invention has the obvious characteristic that the requirements for detection speed and accuracy in the purity detection of a commodity hybrid seed are well satisfied.

Description

technical field [0001] The invention belongs to the field of genetics and discloses a method for quickly and accurately detecting the purity of hybrid rice seeds. Background technique [0002] Seed purity is one of the most important issues in agricultural production. It is related to national food security and is the most important and core content in the development of seed industrialization, so it is also related to the survival of seed companies. The purity requirements of most crop seeds are above 95%, or even 99.9% (Table 1). Therefore, a large amount of statistical sampling is required to obtain accurate data, and the workload is huge. For example, the purity requirement of qualified hybrid rice varieties reaches 96%, which means that only 4 hybrids are allowed in 100 plants, so to make the test results reliable, at least 500 seeds should be tested, and the workload is huge. In addition, due to technical defects, various seed purity detection methods still have probl...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/68
Inventor 彭海张静周俊飞
Owner 海南广陵高科实业有限公司
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