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Potato low-temperature sweetening resistant molecular marker combination and application thereof in potato low-temperature sweetening resistant breeding

A technology of molecular markers and low-temperature saccharification, which is applied in the fields of molecular biology and genetic breeding, can solve the problems of difficult to achieve ideal results and increase the difficulty of breeding, and achieve the effects of easy detection, high sensitivity and good accuracy

Inactive Publication Date: 2016-06-29
HUAZHONG AGRI UNIV
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  • Summary
  • Abstract
  • Description
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  • Application Information

AI Technical Summary

Problems solved by technology

All these studies have shown that low-temperature saccharification of potato tubers is a stable genetic trait, and it is possible to use molecular marker-assisted selection; but because it is controlled by multiple genetic loci, it is difficult to use a single molecular marker similar to quality traits. achieve the desired effect; in addition, potatoes are autotetraploid crops, which will increase the difficulty of conventional selective breeding

Method used

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  • Potato low-temperature sweetening resistant molecular marker combination and application thereof in potato low-temperature sweetening resistant breeding
  • Potato low-temperature sweetening resistant molecular marker combination and application thereof in potato low-temperature sweetening resistant breeding
  • Potato low-temperature sweetening resistant molecular marker combination and application thereof in potato low-temperature sweetening resistant breeding

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0037] 1. Materials: The diploid potato F1 population (EB population) is used as the material. The EB population is 178 offspring produced by crossing the female parent ED25 and the male parent CW2-1. For details, see figure 1 shown. Among them, ED25 contains the blood of two cultivated species S.phureja, S.tuberosum and one wild species S.vernei, which is not resistant to low-temperature saccharification; CW2-1 is a clone of the wild potato species S.berthaultii, which is resistant to low-temperature saccharification.

[0038] The results of phenotypic identification of EB population showed that the reducing sugar content of offspring after low-temperature storage and after warming was significantly separated, showing a normal distribution, which was suitable for QTL mapping analysis.

[0039] 2. Molecular marker experiment method

[0040] 2.1 AFLP markers

[0041] 1) Extraction of genomic DNA

[0042] Genomes were extracted from the young leaves of the EB population using...

Embodiment 2

[0096] Verify the 4 molecular markers screened in Example 1

[0097] The present invention measures the EB population after being planted in 3 different environments (Changling Hill (109.8E, 30.2N), Tianchi Mountain (109.7E, 30.3N) and Wuhan (114.4E, 30.5N)) After the tubers were stored at low temperature ( 4° C. 30 d), the reducing sugar content determination method is the same as that in Example 1; the field experiment design adopts the sequential arrangement method, and each genotype has 3 repetitions, and 10 plants are planted in each repetition.

[0098] Genomes were extracted from young leaves of the EB population using the improved CTAB method, and the specific steps were the same as in Example 1.

[0099] After detection, the content of the extracted genome is 50-200ng / μl, and then the primer pair of S3001-S3004 nucleic acid sequence is used for landing PCR.

[0100] The drop-down PCR system of each molecular marker is 20 μl, specifically as follows: ultrapure water (...

Embodiment 3

[0108] The primer pairs of S3001-S3004 obtained in Example 1 were respectively verified for the broadness of low-temperature glycation resistance.

[0109] 64 potato varieties, breeding parents and breeding high-generation lines with different low-temperature saccharification resistance were selected. The above-mentioned 64 natural population materials were planted in the potato greenhouse of the Huazhong Sub-center of the National Vegetable Improvement Center of Hubei Province. Each material was planted in 4 pots. After the mixed harvest, each material picked 4 larger tubers without pests and diseases and kept them at low temperature ( 4° C.) After storing for 30 days, the reducing sugar content in each tuber was measured by the method in Example 1.

[0110] Genomes were extracted from young potato leaves planted with different materials according to the method in Example 1. The content of the extracted genomes was 50-200 ng / μl, and then landing PCR was performed with primer ...

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Abstract

The invention relates to the fields of molecular biology and genetic breeding and in particular relates to a potato low-temperature sweetening resistant molecular marker and application thereof. Potato low-temperature sweetening resistant molecular marker combination is any one or more of nucleotide sequences S3001-S3004; upstream and downstream primer sequences of the nucleotide sequences S3001-S3004 are shown in SEQ ID No.1-8. The potato low-temperature sweetening resistant molecular marker combination provided by the invention has the beneficial effects that related molecular marker polymorphic analysis and low temperature reducing sugar content determination are carried out on varieties (lines) with different low-temperature sweetening resistant capacities, correlation between the molecular marker and low-temperature sweetening resistance is analyzed, screening is carried out for obtaining a molecular maker combination used for establishing a low-temperature sweetening resistant breeding molecular marker-assisted selection system, the molecular maker combination has relatively high accuracy on low-temperature sweetening resistant potato genotype screening, and technical support is provided for identification of a potato low-temperature sweetening resistant strain and low-temperature sweetening resistant breeding.

Description

technical field [0001] The invention relates to the fields of molecular biology and genetic breeding, in particular to a combination of potato low-temperature saccharification-resistant molecular markers and its application in potato low-temperature saccharification-resistant breeding. Background technique [0002] Potatoes are (Solanum tuberrosum L.) the fourth largest food crop in the world. Potato processing plays an important role in the potato industry. Among them, fried potato products such as potato chips and French fries are popular because of their convenience and unique flavor, accounting for the largest share. However, the frying process has higher requirements on the quality of potato tubers. In addition to the appearance qualities such as large potato pieces, shallow bud eyes, and white potato flesh, the more important thing is that the dry matter content must be above 20%, and the reducing sugar content must be low. At 0.4% (fresh weight). If the reducing suga...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68C12N15/11
CPCC12Q1/6895C12Q2600/13C12Q2600/156
Inventor 宋波涛肖桂林谢从华柳俊黄维曹红菊彭晓君
Owner HUAZHONG AGRI UNIV
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