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Nucleotide primer combination and detection method for rapid identification of the purity of ribbed loofah 'Yalu 6' hybrid seeds

A hybrid seed purity, hybrid seed technology, applied in biochemical equipment and methods, microbial assay/inspection, recombinant DNA technology, etc., can solve problems such as molecular marker identification of hybrid seed purity, achieve accurate and reliable identification results, and reduce identification. Cost and labor saving effect

Active Publication Date: 2019-02-12
肇庆全发农业发展有限公司
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Currently, there is no method for identifying the purity of hybrid seeds using molecular markers in loofah

Method used

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  • Nucleotide primer combination and detection method for rapid identification of the purity of ribbed loofah 'Yalu 6' hybrid seeds
  • Nucleotide primer combination and detection method for rapid identification of the purity of ribbed loofah 'Yalu 6' hybrid seeds
  • Nucleotide primer combination and detection method for rapid identification of the purity of ribbed loofah 'Yalu 6' hybrid seeds

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0029] Example 1 Establishment of the method for detecting the purity of hybrid seeds of ribbed loofah 'Ya Lv No. 6'

[0030] 1. Development of ribbed loofah EST-SSR primers and screening of primers for the purity identification of hybrid seeds of 'Yalu 6'

[0031] Our unit has successfully developed 1046 pairs of loofah EST-SSR primers through loofah transcriptome sequencing. Using these primers, polymorphism screening was carried out on the female parent, male parent and F1 of the 'Yalu 6' variety, and two pairs of co-dominant EST-SSR primer combinations with significant differences between the two parents were obtained, namely SGJ750 and SGJ760, these two pairs of primers have obvious band pattern differences between the two parents, high specificity and good repeatability, and can be used for the purity detection of Yalu 6' first-generation hybrid seeds. The primer sequences are as follows:

[0032] Primer SGJ750

[0033] SGJ750-F: 5'-GATGGCGATAGGGAATCAAA-3' (SEQ ID NO:...

Embodiment 2

[0076] Example 2 Identification of the purity of 50 'Yalu No. 6' F1 plants in the experimental base of the Institute of Vegetables, Guangdong Academy of Agricultural Sciences

[0077] Get 50 ribbed loofah 'Ya Lu No. 6' F1 samples from the experimental base of Vegetable Institute of Guangdong Academy of Agricultural Sciences, number the individual plants, extract genomic DNA, and use the method in Example 1 to identify the purity of hybrid seeds (see image 3 , 4 ), the results showed that among the 50 samples, there were 49 hybrids and 1 female parent, and the seed purity was 98%.

[0078] At the same time, the traditional field observation method was used to identify the hybrid plants for the test, that is, to observe the whole growth period of 50 loofah 'Yalu 6' F1 plants with ribs in the base, and use them with the female and male plants. For comparison, the purity of the hybrid seeds was identified through field observation. The results showed that the molecular marker h...

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Abstract

The invention discloses a nucleotide primer composition for quickly identifying the purity of 'Yalv No.6' hybrid seeds of luffa acutangula roxb and a detection method. In the invention, the purity of hybrid seeds is identified by a molecular marker in the luffa at first, and quick, simple, economical, efficient and accurate purity identification can be realized for the 'Yalv No.6' hybrid seeds of luffa acutangula roxb; and meanwhile, two pairs of primers are adopted for identification, a mutual verification effect can be realized, misjudgment caused by operation error is avoided, and the accuracy of seed purity identification is greatly improved. Compared with traditional field morphological identification, the method only takes 1-3 days from DNA extraction to the end of glue leaking identification, the operation is simple, the labor is saved, land occupation is avoided, the identification result is more accurate and reliable, the identification efficiency is greatly improved, and the identification cost is remarkably reduced. The nucleotide primer composition and the detection method are of great application and promotion value in the enterprises of seed production, breeding and selling of 'Yalv No.6'.

Description

technical field [0001] The invention belongs to the technical field of molecular detection, and in particular relates to a combination of nucleotide primers and a detection method for rapidly identifying the purity of ribbed loofah 'Yalu No. 6' hybrid seeds. Background technique [0002] Hybrid seed purity is an important criterion for measuring seed quality. The impurity of hybrid seeds not only affects crop yields, but also reduces farmers' income. Improper handling may also cause mass incidents and affect social stability. In recent years, incidents caused by hybrid seed impurity have occurred frequently in various crops. Therefore, there is an urgent need to develop related methods for the rapid identification of hybrid seeds in various crops. [0003] For melon vegetable crops, due to the small planting area of ​​ordinary farmers and the small amount of seeds used per unit area, it is easier for farmers to find that hybrid seeds are mixed, which has a greater impact o...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/6895C12N15/11
CPCC12Q1/6895
Inventor 吴海滨罗剑宁龚浩何晓莉郑晓明罗少波张长远何晓明陈俊秋
Owner 肇庆全发农业发展有限公司
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