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A kind of identification between high oleic acid and ordinary oleic acid peanut varieties 1 Non-destructive detection method of seed authenticity
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A damage detection, high oleic acid technology, applied in the field of agricultural bioengineering, can solve the problems affecting seed preservation and germination, seed damage, etc., and achieve the effect of improving breeding efficiency, low cost and reducing workload
Active Publication Date: 2017-09-22
HENAN ACAD OF AGRI SCI
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However, the current methods for detecting these mutations such as CAPS, real-time quantitative PCR, sequencing and AS-PCR all need to extract the DNA of the seeds, which will cause damage to the seeds to varying degrees, and these damages will affect the preservation and germination of the seeds; and not all breeding methods Institutions have the experimental conditions to detect these mutations by means of molecular biology
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Embodiment 1
[0025] Embodiment 1. A non-destructive detection method for distinguishing the authenticity of F1 seeds between high oleic acid and common oleic acid peanut varieties, comprising the following steps:
[0026] (1) Harvest all F from the hybrid combination between high oleic acid and normal oleic acid 1 Seeds and seeds selfed by the female parent under the same planting conditions;
[0027] (2) Using near-infrared spectroscopy to measure the female parent and F of a single grain, respectively 1 Oleic acid content of seeds. The female parent measures the oleic acid content of 10-20 seeds, and gets the average value for later use;
[0028] Steps for determining the content of oleic acid by near-infrared spectroscopy: Turn on the power of the near-infrared analyzer (DA7200 DiodeArray Analyzer) and preheat it for half an hour; Analyze the spectral curve, click Measure, save the reading, and repeat three times.
[0029] (3) When the female parent is common oleic acid peanut, calc...
Embodiment 2
[0033] Example 2: Using molecular markers to identify true and false hybrids in hybrid seeds.
[0034] Harvest the hybrid F of the hybrid combination wt09-0023×Yuanza 9102 and Yuanza 9102×wt09-0023 respectively 1 seeds, for each F 1 Seed numbering, a small amount of tissue was cut from the far embryonic end of the seed to extract DNA. The FAD2 gene of Yuanza 9102 (ah FAD2A and ah FAD2B ) genotype is AABB, and the genotype of the FAD2 gene of wt09-0023 is aabb, so the genotype of the FAD2 gene of the above two combinations of true hybrid seeds is AaBb; The type is aabb, and the genotype of the pseudo-hybrid in the combination of 9102×wt09-0023 is AABB.
[0035] Each F can be obtained by sequencing 1 The genotype of the seed, which distinguishes true and false hybrids based on genotype. F 1Seed DNA was used as a template, and PCR was performed with primers aF19 / R3 and bF19 / R3 (Patel, 2004), and genotypes could be distinguished according to the sequencing peaks of PCR pro...
Embodiment 3
[0038] Embodiment 3, the comparison of near-infrared spectroscopy and gas chromatography to measure the content of oleic acid in peanut seeds
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Abstract
The invention relates to a nondestructive detection method for identifying F1 seed true / false between species of peanut with a high oleic acid content and peanut a with normal oleic acid content. The method comprises the following steps: harvesting all F1 seeds, which are obtained by hybridizing species of peanut with a high oleic acid content and peanut a with normal oleic acid content and seeds of female selfing under same planting conditions; individually measuring the oleic acid contents of single granules of mother parent and F1 by a near infrared spectrum technology; when the female parent is peanut with a common oleic acid content, calculating the ratio of F1 seeds that contains a oleic acid content higher than that of female parent, if the ratio is more than 10%, judging the F1 seeds are true hybrids, if the ratio is less than 10%, judging the F1 seeds are false hybrids; when the female parent is peanut with a high oleic acid content, calculating the ratio of F1 seeds that contains a oleic acid content lower than that of female parent, if the ratio is more than 20%, judging the F1 seeds are true hybrids, and if the ratio is less than 20%, judging the F1 seeds are false hybrids. The provided method has the advantages that the seed are not damaged, no pretreatment is needed, the method is pollution-free, fast, and convenient, the detection speed is quick; the detection only needs a near-infrared spectrum analyzer, and the operation is convenient and quick, and is easy to apply.
Description
technical field [0001] The invention belongs to the field of agricultural bioengineering, and specifically relates to a method for distinguishing between high oleic acid and common oleic acid peanut varieties. 1 Non-destructive detection method for seed authenticity. Background technique [0002] The composition and content of fatty acids in oil crop seeds are important indicators to measure their quality. In the fatty acid composition of peanut seeds, the content of oleic acid and linoleic acid is about 80%. Oleic acid has good oxidation stability and can reduce lipid oxidation. If the content of oleic acid in peanut fatty acid is increased, the shortcomings such as bad smell, taste and shortened shelf life due to lipid oxidation can be improved. High oleic acid peanuts are beneficial to human health. Eating high oleic acid peanuts is beneficial to reduce the risk of cardiovascular and cerebrovascular diseases, and can also reversely inhibit human inflammatory cytokine TN...
Claims
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