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Method for identifying hybrid seeds of winter cole-wort and mustard type winter rape

A technology of winter rapeseed and mustard type, applied in the field of identification of brassica napus and mustard type winter rapeseed hybrids, can solve the problem of rare seed purity

Inactive Publication Date: 2015-05-13
GANSU AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although traditional field morphological identification is still the most commonly used method for identification of variety authenticity and seed purity, it is relatively rare to use peroxide isoenzymes to identify variety authenticity and seed purity.

Method used

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  • Method for identifying hybrid seeds of winter cole-wort and mustard type winter rape
  • Method for identifying hybrid seeds of winter cole-wort and mustard type winter rape
  • Method for identifying hybrid seeds of winter cole-wort and mustard type winter rape

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0040] Embodiment 1: a kind of method of identification brassica napus and mustard type winter rape hybrid comprises the following steps:

[0041] (1) Sampling: Pick flower buds, flowers, and siliques of rape in the field from April to June every year, put them in an ice box within 3-5 minutes, bring them back to the laboratory, and store them in a 4°C refrigerator for later use;

[0042] (2) After weighing 0.2-0.3g flower buds, flowers, and siliques, respectively, use pH 7.8 phosphate buffer solution containing 2mmolASA, 5mmolEDTA-Na2 to homogenize in an ice bath, centrifuge at 12000r / min for 10min, and take the The supernatant was stored in a -20°C refrigerator for later use. Using polyacrylamide gel electrophoresis, the concentrations of the separating gel and stacking gel were 7% and 4%, respectively, and the pH values ​​were 8.8 and 6.8 respectively; the electrode buffer used Tris-glycine system, the pH value was 8.3, and the sample volume was 30 μL; At the beginning, th...

Embodiment 2

[0044] Embodiment 2: a kind of method of identification brassica napus and mustard type winter rape hybrid comprises the following steps:

[0045](1) Sampling: Pick flower buds, flowers, and siliques of rape in the field from April to June every year, put them in an ice box within 3-5 minutes, bring them back to the laboratory, and store them in a 4°C refrigerator for later use;

[0046] (2) After weighing 0.2-0.3g flower buds, flowers, and siliques, respectively, use pH 7.8 phosphate buffer solution containing 2mmolASA, 5mmolEDTA-Na2 to homogenize in an ice bath, centrifuge at 12000r / min for 10min, and take the The supernatant was stored in a -20°C refrigerator for later use. Using polyacrylamide gel electrophoresis, the concentrations of the separating gel and stacking gel were 7% and 4%, respectively, and the pH values ​​were 8.8 and 6.8 respectively; the electrode buffer used Tris-glycine system, the pH value was 8.3, and the sample volume was 30 μL; At the beginning, the...

Embodiment 3

[0051] Embodiment 3: a kind of method of identification brassica napus and mustard type winter rape hybrid comprises the following steps:

[0052] Steps (1), (2), (3), and (4) are the same as above.

[0053] The POD isozyme analysis of buds, flowers, and siliques of mustard-sugar hybrid progenies and their parents showed that (Fig. Although the POD isoenzyme bands in siliques were consistent, the corresponding enzyme bands were significantly different in depth of staining, and those with darker staining bands had relatively stronger activity. Strong, which is consistent with the results measured by physiological and biochemical indicators; the POD isoenzyme band of Shanxi winter mustard x vision is completely different from that of the parent, and it also has the band of the parent parent, and there is an additional enzyme band with an Rf value of 0.145.

[0054] Figure 11(a) and (b) illustrate that the POD isozyme analysis of the flower buds, flowers and siliques of mustard-...

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Abstract

The invention relates to a method for identifying hybrid seeds of winter cole-wort and mustard type winter rape. With the adoption of the method, the condition that the hybrid seeds of winter cole-wort and mustard type winter rape are real hybrid seeds is identified. The method for identifying the hybrid seeds of winter cole-wort and mustard type winter rape is characterized by comprising steps as follows: (1), sampling; (2), weighing 0.2-0.3 g of flower buds, flowers and silique respectively, performing homogenizing by adopting a phosphate buffer having the pH of 7.8 and containing 2 mmol of ASA (acrylonitrile styrene acrylate) and 5 mmol of EDTA-Na2 (ethylene diamine tetraacetic acid-Na2) under ice-bath respectively, performing centrifugation for 10 min at 12,000 r / min, and taking a liquid supernatant and storing the liquid supernatant in a refrigerator at the temperature of subzero 20 DEG C for standby. Polyacrylamide gel electrophoresis is adopted, concentrations of separation gel and spacer gel are 7% and 4% respectively, and pH values are 8.8 and 6.8 respectively; an electrode buffer adopts a Tris-glycine system, the pH value is 8.3, and the loading quantity of samples is 30 mu L; when electrophoresis starts, steady voltage is 100 V, the voltage is regulated to be 200 V after the samples pass the concentrated gel, and the electrophoresis time is about 5 h; after electrophoresis is finished, the samples are placed in a dyeing liquid benzidine solution for dyeing until a clear blue stripe can be observed, and the samples are photographed for storage after washed by flowing water; a migration rate Rf value is quotient obtained by dividing the migration distance from an original point to the stripe by the distance from the original point to an indicator.

Description

technical field [0001] The invention relates to a method for identifying hybrids of Brassica napus and Brassica napus hybrids. Using this method, the hybrids of Brassica napus and Brassica napus were successfully identified as true hybrids. Background technique [0002] Distant hybridization refers to the hybridization between different species, genus or species with more distant relationship, including interspecific hybridization, intergeneric hybridization and inter-subspecies hybridization. Distant hybridization is an important means of breeding and an important way to create new types of plants and new varieties of crops. Through distant hybridization, the boundaries between species and genus can be broken, the exchange or combination of genetic material between different species can be made possible, and plant traits can be improved purposefully to form new species and varieties with stronger vitality. According to whether the newly formed species completely contain t...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/28
Inventor 刘自刚钱武孙万仓李学才方彦武军艳杨刚王月王凯音刘林波侯献飞马骊张树娟杨建胜刘海卿董红叶
Owner GANSU AGRI UNIV
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