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Thin-shell hickory variety Moore characteristic sequences, marker primers and identification method

A technology of pecans with thin shell and marker primers, which is applied in biochemical equipment and methods, microbial determination/inspection, DNA/RNA fragments, etc., can solve instability, low yield, inconvenient variety identification, promotion of new variety cultivation, etc. problem, to achieve the effect of improved reliability and simple method

Active Publication Date: 2018-01-16
ZHEJIANG FORESTRY ACAD
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0003] However, the main problem faced by the existing pecan production areas in my country is low yield and instability. There are many reasons for this phenomenon, one of which is the lack of clear genetic relationship analysis of the existing introduced varieties Coupled with the confusion in the naming of some hybrid offspring, it is difficult to carry out effective parent selection and rational allocation, and it is not convenient for variety identification, promotion, communication, and cultivation of new varieties. Therefore, efforts have been made to develop stable and specific DNA fingerprints for these varieties at the molecular level. Marking is the scientific way to realize accurate and rapid identification of pecan varieties
[0004] The identification and genetic relationship analysis of pecan varieties based on SSR molecular markers have been reported at home and abroad, but the detection is cumbersome and the results are unstable

Method used

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  • Thin-shell hickory variety Moore characteristic sequences, marker primers and identification method
  • Thin-shell hickory variety Moore characteristic sequences, marker primers and identification method
  • Thin-shell hickory variety Moore characteristic sequences, marker primers and identification method

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Embodiment 1

[0038] (1) Extraction of Genomic DNA of Thin-shelled Hickory Nut Variety:

[0039] Take 0.01g of the young leaves of the pecan variety to be tested, add liquid nitrogen and grind them thoroughly, and use the SDS-CTAB method to extract the genomic DNA. thing. The crude DNA was purified by Magabio Nucleic Acid Purification Kit (Bioer, Hangzhou, China), and detected by 1.5% agarose gel electrophoresis and DNA / RNA UV spectrophotometer (GeneQuant Pro, GE Healthcare). Integrity, Purity and Concentration. DNA samples with OD260 / OD280>1.8 were used for subsequent PCR amplification. DNA extracts were stored in a -20°C refrigerator for later use.

[0040] (2) Design specific PCR amplification primers, the sequence of the primer pair is:

[0041]The upstream primer: 5'-TGTCAAGGACAAACGGGTGA-3'; and the downstream primer: 5'-TGGAGAAGCGAAGTAGTCAACC-3' were synthesized by Shanghai Bioengineering Technology Co., Ltd.

[0042] (4) PCR amplification:

[0043] Composition of PCR reaction s...

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Abstract

The invention relates to a pair of high-specificity molecular-specificity marker primers for a thin-shell hickory variety Moore, and a method for rapid identification of the thin-shell hickory varietyMoore. Primer sequences comprise the following sequences: an upstream primer: 5'-TGTCAAGGACAAACGGGTGA-3'; and a downstream primer: 5'-TGGAGAAGCGAAGTAGTCAACC-3'. The molecular-specificity marker primers can be used for rapid early identification of the thin-shell hickory variety Moore. The method is simple, rapid and accurate, and is an irreplaceable molecular method to identify the thin-shell hickory variety with apparent characteristics.

Description

(1) Technical field [0001] The invention relates to a characteristic sequence of the pecan variety Moore, a molecular specific marker primer, and a method for rapidly identifying the pecan variety Moore by using the molecular specific marker primer. (2) Background technology [0002] Shelled Pecans (Carya (Wangenh.) K.Koch) is the most economically valuable tree species in the Juglandaceae Hickory genus. Carya carya is a typical outcrossing plant. According to the existing production practice, the variety configuration of carya carya is one of the main factors affecting its yield. The U.S. is one of the origins of pecans and its central production area. Over the years, it has selected and named about 1,000 cultivars. It has been more than 100 years since the introduction of hickory nuts in my country, and there are dozens of varieties commonly used in production at present. Years of production practice have proved that the vast subtropical regions of my country are suitab...

Claims

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Application Information

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IPC IPC(8): C12Q1/6895C12N15/11
Inventor 彭华正朱汤军金群英叶华琳
Owner ZHEJIANG FORESTRY ACAD