CN-121006418-B - Application of Indel marking dosage typing in detection of sweet potato soft rot resistance

Abstract

The invention discloses an application of Indel marking dosage typing in detecting sweet potato soft rot resistance, belonging to the technical field of plant genetic engineering. According to the invention, the Indel molecular marker related to sweet potato soft rot resistance is obtained through screening, the nucleotide sequence of the Indel molecular marker is shown as SEQ ID NO.2, and the sequence shown as SEQ ID NO.2 is inserted into or deleted from 161 th site of the sequence 8 bp. Experiments show that the target band of the amplified product is obtained through amplification of Indel molecular markers and electrophoresis, dosage values of two genotypes are obtained according to gray values of the dyed electrophoretic band, and the result shows that the inserted Indel molecular markers are positively correlated with the sweet potato soft rot resistance, so that the method has important significance for dose effect research and molecular assisted breeding of the sweet potato gene functions.

Inventors

• ZHANG HUAN
• HE SHAOZHEN
• LIU QINGCHANG
• ZHAI HONG
• GAO SHAOPEI
• ZHAO NING
• CHEN PEITAO

Assignees

• 中国农业大学

Dates

Publication Date

20260505

Application Date

20251027

Claims (7)

1. 1. Use of a detected Indel molecular labelled reagent in any of the following: (1) The application in detecting the soft rot resistance of sweet potatoes; (2) Application in sweet potato soft rot resistance molecular breeding; (3) Application in screening sweet potato with soft rot resistance; The Indel molecular marker nucleotide sequence is shown as SEQ ID NO.2, and the sequence shown as SEQ ID NO.2 is inserted into or deleted from 161 th site by 8bp sequence; The 8bp sequence of the insertion or deletion is TTGGTCAG; The dosage of the inserted Indel molecular marker is positively correlated with the sweet potato soft rot resistance.
2. 2. The use according to claim 1, wherein the dosage of the inserted Indel molecular marker is calculated from the number of homologous chromosomes of the inserted Indel molecular marker in the homologous hexaploid sweet potato chromosome.
3. 3. Use of a primer pair for detecting Indel molecular markers or a kit comprising said primer pair in any of the following: (1) The application in detecting the soft rot resistance of sweet potatoes; (2) Application in sweet potato soft rot resistance molecular breeding; (3) Application in screening sweet potato with soft rot resistance; The Indel molecular marker nucleotide sequence is shown as SEQ ID NO.2, and the sequence shown as SEQ ID NO.2 is inserted into or deleted from 161 th site by 8bp sequence; The 8bp sequence of the insertion or deletion is TTGGTCAG; The dosage of the inserted Indel molecular marker is positively correlated with the sweet potato soft rot resistance.
4. 4. The use according to claim 3, wherein the nucleotide sequence of the primer pair is shown in SEQ ID No. 3-4.
5. 5. The use according to claim 3, wherein the dosage of the inserted Indel molecular marker is calculated from the number of homologous chromosomes of the inserted Indel molecular marker in the homologous hexaploid sweet potato chromosome.
6. 6. A method for detecting soft rot resistance of sweet potatoes, comprising the steps of: Taking genomic DNA of the sweet potato to be detected as a template, utilizing a primer pair with a nucleotide sequence shown as SEQ ID NO.3-4 to amplify Indel molecular markers, carrying out electrophoresis on amplified products, obtaining dosage values of two genotypes according to gray values after electrophoresis band dyeing, and judging soft rot resistance of the sweet potato to be detected according to the dosage values of the two genotypes; The Indel molecular marker nucleotide sequence is shown as SEQ ID NO.2, and the sequence shown as SEQ ID NO.2 is inserted into or deleted from 161 th site by 8bp sequence; The 8bp sequence of the insertion or deletion is TTGGTCAG; the judging method is that the dosage of the inserted Indel molecular marker is positively correlated with the soft rot resistance of the sweet potato.
7. 7. The method of claim 6, wherein the dosage values of the two genotypes are calculated from the number of homologous chromosomes in the homologous hexaploid sweet potato chromosome based on the ratio of Indel molecular markers of the two different genotypes.

Description

Application of Indel marking dosage typing in detection of sweet potato soft rot resistance Technical Field The invention relates to the technical field of plant genetic engineering, in particular to application of Indel marker dosage typing in detection of sweet potato soft rot resistance. Background Sweet potato (Ipomoea batatas [ L. ] lam.) is a hexaploid crop (2n=6x=90), which has been domesticated for over 5000 years in south america. The sweet potato has high yield and strong environmental compliance, contains a large amount of starch, and can be used as important grains, feed, industrial raw materials and novel energy. Sweet potato is an excellent source of nutrients including vitamins, potassium, iron, calcium and minerals, and has medicinal value due to its anticancer, antidiabetic and antiinflammatory activities, and is highly favored by people. There are a large number of Indel allelic variations in the sweetpotato genome, which often lead to changes in gene expression levels, regulatory functions, translation results. Indels cannot be typed by conventional methods such as PCR-agarose electrophoresis due to the complex genome and polyploidy of sweet potato. The gene expression and regulation level difference caused by Indel allelic variation cannot be estimated, and the development of gene function research of sweet potatoes is hindered. Dose effects cannot be distinguished using conventional agarose electrophoresis, and are expensive to distinguish using high throughput sequencing. Therefore, developing a dose typing method suitable for homologous hexaploid sweet potato Indel is important for sweet potato gene function research. Disclosure of Invention The invention aims to provide application of Indel marker dosage typing in detecting sweet potato soft rot resistance, so as to solve the problems in the prior art, obtain corresponding gray values of two genotype Indel molecular marker strips through high-concentration polyacrylamide gel electrophoresis, calculate the dosage of Indel molecular markers according to gray value proportion, find that the dosage of insert Indel molecular markers is positively correlated with sweet potato soft rot resistance, and have important significance for dose effect research and molecular assisted breeding of sweet potato gene functions. In order to achieve the above object, the present invention provides the following solutions: the invention provides an Indel molecular marker related to sweet potato soft rot resistance, the nucleotide sequence of the Indel molecular marker is shown as SEQ ID NO.2, and the sequence shown as SEQ ID NO.2 is inserted into or deleted from 161 th site by 8bp sequence. Preferably, the 8bp sequence of the insertion or deletion is TTGGTCAG. The invention also provides a primer pair for amplifying the Indel molecular marker, and the nucleotide sequence of the primer pair is shown as SEQ ID NO. 3-4. The invention also provides a kit for detecting the resistance related to the sweet potato soft rot, which comprises the primer pair. The invention also provides the application of the Indel molecular marker or the primer pair or the kit in any one of the following: (1) The application in detecting the soft rot resistance of sweet potatoes; (2) Application in sweet potato soft rot resistance molecular breeding; (3) The application in screening sweet potatoes with soft rot resistance. Preferably, the dosage of the inserted Indel molecular marker is positively correlated with sweet potato soft rot resistance. Preferably, the dosage of the inserted Indel molecular marker is calculated according to the number of homologous chromosomes of the insertion Indel molecular marker in the homologous hexaploid sweet potato chromosome. The invention also provides a method for detecting the soft rot resistance of the sweet potato, which comprises the following steps: And (3) taking genomic DNA of the sweet potato to be detected as a template, amplifying the Indel molecular marker by using a primer pair with a nucleotide sequence shown as SEQ ID NO.3-4, carrying out electrophoresis on an amplified product, obtaining dosage values of two genotypes according to the gray value after dyeing of an electrophoresis strip, and judging the soft rot resistance of the sweet potato to be detected according to the dosage values of the two genotypes. Preferably, the dosage values of the two genotypes are calculated according to the number of homologous chromosomes of the Indel molecular markers of the two different genotypes, wherein the number of homologous chromosomes is the ratio of the homologous hexaploid sweet potato chromosomes. Preferably, the judgment method is that the dosage of the inserted Indel molecular marker is positively correlated with the soft rot resistance of the sweet potato. The invention discloses the following technical effects: The invention provides a dosage typing method suitable for homologous hexaploid sweet potato indels for the first time,