InDel molecular marker related to character of gillyflower leaf hair and application of InDel molecular marker

Abstract

The invention discloses an InDel molecular marker related to violet leaf hair traits and application of the InDel molecular marker, primers of the InDel molecular marker can specifically amplify strips related to the violet leaf hair traits, and the violet leaf hair traits can be accurately judged by extracting to-be-detected violet genome DNA (deoxyribonucleic acid), performing PCR (polymerase chain reaction) amplification by the primers and detecting amplification products. The molecular marker has the characteristics of codominance, strong specificity and good stability, can be widely applied to the fields of germplasm resource screening, classification, assisted breeding and the like of the violet, and provides powerful technical support for relevant research and industrial development of the violet.

Claims

1. An InDel molecular marker associated with the pubescent trait of violet leaves, characterized in that, The InDel molecule is labeled InDel-MNSM1, and the nucleotide sequence of InDel-MNSM1 is shown in SEQ ID NO:1; The sequence shown in SEQ ID NO:1 is missing in the genome of the leafy violet. The genome of the leafless pubescent violet contains a sequence as shown in SEQ ID NO:

1.

2. The amplification primers for the InDel molecular marker associated with the pubescent trait of violet leaves as described in claim 1, characterized in that, The amplification primers are MiFP-SM, and the nucleotide sequence of MiFP-SM is as follows: Upstream primer: 5'-AGGTCAGATTCAAGAACAGAGT-3'; Downstream primer: 5'-GATTCATCCGAGGAACATCAA-3'.

3. The application of the InDel molecular markers associated with the pubescent trait of violets as described in claim 1 or the amplification primers of the InDel molecular markers associated with the pubescent trait of violets as described in claim 2 in the identification of the pubescent trait of violets or in violet breeding.

4. The application according to claim 3, characterized in that, The test detects whether the genomic DNA of the violet to be tested contains the sequence shown in SEQ ID NO:1; if it contains the sequence, the violet to be tested is determined to be a leafless hairy violet, otherwise it is determined to be a leafy hairy violet.

5. The application according to claim 4, characterized in that, Using the genomic DNA of the violet plant to be tested as a template, PCR amplification was performed using the amplification primers shown in SEQ ID NO:2 and SEQ ID NO:

3. The PCR amplification products were separated by electrophoresis, and the genotype was determined based on the size of the bands. When the amplification product is a single band of 260bp, the violet to be tested is a leafy, hairy violet. When the amplification product is a single band of 741bp, the violet to be tested is *Viola yedoensis*. When the amplification product is a double band of 260bp and 741bp, the violet to be tested is a leafy, hairy violet.

6. The application according to claim 4, characterized in that, The PCR amplification reaction conditions were as follows: 94℃ pre-denaturation for 5 min; 94℃ denaturation for 30 s, 58℃ annealing for 30 s, 72℃ extension for 45 s, for a total of 35 cycles; 72℃ final extension for 10 min.

7. The application according to claim 4, characterized in that, The PCR amplification was performed using a 20 μL PCR reaction system, comprising: 2 μL of 10×PCR Buffer, 1.6 μL of 2.5 mmol / L dNTPs, 0.8 μL of 10 μmol / L upstream primer, 0.8 μL of 10 μmol / L downstream primer, 0.2 μL of 5 U / μL Taq DNA polymerase, 1 μL of 50 ng / μL template DNA, and ddH2O to a final volume of 20 μL.

8. A method for breeding violets based on the pubescent trait of violet leaves, characterized in that, include: Using the genomic DNA of the violet to be tested as a template, PCR amplification was performed using the amplification primers as described in claim 2. Based on the amplification products, violet germplasm materials with the target leaf hairy trait were screened out, and hairy or hairless phenotype single plants were selectively retained.

9. A kit for detecting the pubescence trait of violet leaves, characterized in that, Includes the amplification primers as described in claim 2.

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