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A molecular marker and its application for identifying the purity of Indian pumpkin 'Yinhui No. 3' hybrid seeds

A hybrid seed purity, Indian pumpkin technology, applied in the determination/inspection of microorganisms, biochemical equipment and methods, DNA/RNA fragments, etc., to achieve the effects of good specificity, high commercial application value, and high stability

Inactive Publication Date: 2020-12-22
NORTHEAST AGRICULTURAL UNIVERSITY
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AI Technical Summary

Problems solved by technology

In the prior art, there is still no method for identifying the purity of Indian pumpkin 'Yinhui No. 3' hybrid seeds. It is urgent to establish a set of stable, reliable, simple and fast Indian pumpkin 'Yinhui No. 3' seed purity testing methods as soon as possible.

Method used

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  • A molecular marker and its application for identifying the purity of Indian pumpkin 'Yinhui No. 3' hybrid seeds
  • A molecular marker and its application for identifying the purity of Indian pumpkin 'Yinhui No. 3' hybrid seeds
  • A molecular marker and its application for identifying the purity of Indian pumpkin 'Yinhui No. 3' hybrid seeds

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Embodiment 1

[0033] 1. Extraction of Pumpkin Genomic DNA

[0034] Planting Parents and Hybrid F 1 When the true leaves unfolded, the total genomic DNA of the leaves was extracted by the CTAB method. Take a newly unfolded true leaf into a 1.5mL centrifuge tube and add liquid nitrogen, grind the tissue into powder, add 700μL CTAB lysis buffer, shake vigorously for 30 seconds, bathe in 60°C water for 30 minutes, and mix well by inverting up and down during the period. Add 700 μL of chloroform isoamyl alcohol (24:1, v / v) to the above homogenized lysate, mix well by inverting up and down, and centrifuge at 13000 r / min at 4°C for 15 minutes. Pipette 400 μL of the supernatant into a new centrifuge tube, add an equal volume of isopropanol, mix well by inverting up and down, let stand at -20°C for 30 minutes, and centrifuge at 12,000 r / min at 4°C for 10 minutes. Discard the supernatant, add 200 μL of 75% ethanol along the wall of the centrifuge tube, wash the tube wall upside down, centrifuge at ...

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Abstract

The invention discloses a molecular marker for identifying the purity of a hybrid seed of winter squash Yinhui No.3 and applications thereof, and belongs to the technical field of molecular breeding. The primer pair for PCR amplified molecular marking is Yinhui3-F and Yinhui3-R. The nucleotide sequence of Yinhui3-F is represented by SEQ ID No.3, and the nucleotide sequence of Yinhui3-R is represented by SEQ ID No.4. If through the primer pair, a 91bp sequence and an 85be sequence can be specifically amplified out at the same time, the hybrid F1 plant is a real hybrid species. If any one of the 91bp sequence and 85be sequence is not amplified out, the hybrid F1 plant is a fake hybrid species. The invention also provides a method for identifying the purity of a pumpkin hybrid seed. The provided SSR molecular marker and primers have high stability, and can be simply and rapidly applied to the molecular marker assisted breeding and identify the purity of a hybrid seed of winter squash Yinhui No.3.

Description

technical field [0001] The invention relates to a molecular marker for identifying the purity of Indian pumpkin 'Yinhui No. 3' hybrid seeds and an application thereof, belonging to the technical field of molecular breeding. Background technique [0002] Pumpkin (2n=2x=40) (Cucurbita spp.) belongs to Cucurbitaceae (Cucurbitaceae) Cucurbita spp., is an annual vine herb, and is an important worldwide vegetable. Pumpkin has various uses, has high nutritional and medicinal value, and is deeply loved by consumers. In recent years, breeders have bred a number of new pumpkin varieties according to the needs of consumers. The Indian pumpkin 'Yinhui No. 3' hybrid cultivated by the inventor's pumpkin breeding laboratory has high yield, white grains, and a high seed yield. High and disease-resistant, it is deeply loved by growers in production. [0003] 'Yinhui No.3' is a first-generation pumpkin variety hybridized between the female parent '088' and the male parent '9-6'. During seed...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/6895C12N15/11
CPCC12Q1/6895C12Q2600/156
Inventor 屈淑平王云莉闫春冬王洋洋徐文龙
Owner NORTHEAST AGRICULTURAL UNIVERSITY
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