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TRAP (Target Region Amplification Polymorphism) marker primer combination for genetic diversity analysis on wild sibling species of Sinkiang amaryllidaceae and application of TRAP marker primer combination

A technique for genetic diversity and labeled primers, applied in the field of TRAP primer combinations

Active Publication Date: 2018-11-13
XINJIANG AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

TRAP molecular markers have the characteristics of simple operation, good reproducibility, high efficiency, and good stability. Allium cepa , L.) genotypes. Scientia Horticulturae , 2015, 190: 123-127), short-day onion germplasm identification was successfully applied (Kisha TJ, Cramer CS. Determining redundancy of Short-day Onion accessions in agermplasm collection using Microsatellite and Targeted Region Amplified Polymorphic Markers. Journal of the American Society for Horticultural Science American Society , 2011, 136(2): 129-134), but there are no reports on the construction of the genetic map of Allium plants, the markers of important traits, the cloning of related genes, etc., especially the combination of TRAP marker primers and the application of this The analysis of genetic diversity of wild relatives of onion and garlic in Xinjiang has not been reported yet

Method used

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  • TRAP (Target Region Amplification Polymorphism) marker primer combination for genetic diversity analysis on wild sibling species of Sinkiang amaryllidaceae and application of TRAP marker primer combination
  • TRAP (Target Region Amplification Polymorphism) marker primer combination for genetic diversity analysis on wild sibling species of Sinkiang amaryllidaceae and application of TRAP marker primer combination
  • TRAP (Target Region Amplification Polymorphism) marker primer combination for genetic diversity analysis on wild sibling species of Sinkiang amaryllidaceae and application of TRAP marker primer combination

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0027] Example 1. TRAP primer design

[0028] Search the EST sequences of Allium plants from the NCBI database, design primers based on their gene sequences, and design 3 fixed primers (FY-F1, FS-F1, FS-F2) and 3 random primers (FY-R1, FS- R1, FS-R2).

Embodiment 2

[0029] Example 2 Analysis of the genetic diversity of wild relatives of onion and garlic in Xinjiang by using the combination of TRAP molecular marker primers includes the following steps:

[0030] (1) Material

[0031] Altai garlic, Xinjiang garlic, healthy garlic, multi-seed garlic, Qitai garlic, Kanas garlic, Hongqi Daban garlic, edge-leaved garlic, scallion and small mountain garlic are selected in the original environment, and a total of 10 wild relatives of Xinjiang onion and garlic are selected. The species is the experimental material, and the sample material for TRAP analysis is taken from fresh leaves ( figure 1 ), the silica gel quickly dries the sample. The details of the source of materials are shown in Table 1.

[0032]

[0033] (2) Genomic DNA was extracted from wild relatives of onion and garlic in Xinjiang by the improved SDS method.

[0034] Using the mixed DNA of samples from different populations as the template DNA, 9 pairs of TRAP primer combinat...

experiment example 2

[0049] Experimental Example 2 Summary

[0050] Using the primer combination provided by the present invention, the genetic diversity analysis of Xinjiang onion and garlic wild relatives was realized by TRAP-PCR amplification, in order to realize the taxonomic identification and genetic relationship of Xinjiang onion and garlic wild relatives based on TRAP molecular markers Identification and other technical support were provided, and the efficiency and applicability of the TRAP primer combination in the study of wild relatives of onion and garlic in Xinjiang were further elaborated.

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Abstract

The invention belongs to the field of molecular biology DNA (Deoxyribonucleic Acid) marking techniques and application and discloses a TRAP (Target Region Amplification Polymorphism) marker primer combination for genetic diversity analysis on wild sibling species of Sinkiang amaryllidaceae and the application of the TRAP marker primer combination. The primer combination provided by the invention has a nucleotide sequence of SEQ ID No.1 to SEQ ID No.6 as shown in the specification. A TRAP-PCR (Polymerase Chain Reaction) system provided by the invention has a dNTPs (Deoxynucleotide) concentration of 0.250mmol / L, a primer concentration of 0.250mu mol / L, a Taq enzyme amount of 0.750 U and a template DNA amount of 75.000ng, the obtained primer combination which is clear in strip, good in repeatability and high in polymorphism is capable of accurately and reliably carrying out genetic analysis, and due to the high efficiency and the practicability of the primer combination provided by the invention, the application of TRAP markers to study on wild sibling species of Sinkiang amaryllidaceae is made up.

Description

technical field [0001] The invention belongs to the field of DNA marker technology and application of molecular biology, and in particular relates to a TRAP primer combination for analyzing the genetic diversity of wild relatives of onion and garlic in Xinjiang and its application. Background technique [0002] Onion and garlic plants belong to the genus Allium (Amarylllidaceae) according to APG IV. Allium L.) Herbaceous perennials. The wild relatives of garlic, leeks, scallions and onions are stored in mountainous forests and grasslands, foothill wastelands, arid deserts and arid grassland shrubs in the north of the Tianshan Mountains, south of the Altai Mountains, and north of the Kunlun Mountains in Xinjiang. Due to the special geographical environment, some wild relatives of onion and garlic plants are only distributed in Xinjiang in China, which are not only important ornamental, edible, medicinal, vegetable and rearing plant resources, but also valuable resources for...

Claims

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Application Information

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IPC IPC(8): C12Q1/6895C12N15/11
CPCC12Q1/6895C12Q2600/156
Inventor 林辰壹王丹丹
Owner XINJIANG AGRI UNIV