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Specific marker primers for sakura varieties including Plena Rosea, Sakuyahime and Ujou-shidare

A technology of eight-layer red branch weeping and labeling primers, which is applied in the direction of recombinant DNA technology, microbial measurement/inspection, biochemical equipment and methods, etc., and can solve the difficult identification of phenotypic characteristics, the application and promotion of garden plants, and the selection of new varieties Difficulties and other problems, to achieve the effect of simple methods

Active Publication Date: 2015-05-13
ZHEJIANG FORESTRY ACAD
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Problems solved by technology

After long-term investigation work, we found that 20 late cherry varieties include Hongli (Cerasus serrulata'Benigasa'), Honghua (C.serrulata'Kouka'), Yiye (C.serrulata'Hisakura'), Fulushou ( C.serrulata'Contorta'), Samantabhadra (C.serrulata'Albo-rosea'), Songyue (C.serrulata'Superba'), Yujin (C.serrulata'Grandifora'), Hongfeng (Cerasus×sieboldii' Beni‐yutaka'), Royal Clothes Yellow (C.serrulata'Gioiko'), Guanshan (C.serrulata'Kanzan'), Lantern (C.serrulata'Ojochin'), Eightfold Red Branch Weeping (C.spachiana'Plena Rosea' ), Ichihara Huwei (C.serrulata'Albo Plena'), Sakuyahime (Cerasus×yedoensis'Sakuyahime'), Suzaku (C.serrulata'Shujaku'), Yang Guifei (C.serrulata'Mollis'), Yuqing Zhiwei (C.spachiana'Ujou‐shidare'), Chrysanthemum weeping cherry (C.serrulata'Plena-pendula'), Hirano sister back (C.serrulata'Imose'), Asahiyama cherry (C.serrulata'Asahiyama') in the garden It is widely used, but the phenomenon of "same names and different objects" and "same objects and different names" of these varieties is also very serious, and it is difficult to identify them from the aspect of phenotypic characteristics, which brings many difficulties to the application, promotion and selection of new varieties of garden plants. Therefore, Focusing on the development of stable and specific DNA fingerprints for these varieties at the molecular level is the scientific way to achieve accurate and rapid identification of cherry blossom varieties

Method used

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  • Specific marker primers for sakura varieties including Plena Rosea, Sakuyahime and Ujou-shidare
  • Specific marker primers for sakura varieties including Plena Rosea, Sakuyahime and Ujou-shidare
  • Specific marker primers for sakura varieties including Plena Rosea, Sakuyahime and Ujou-shidare

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Embodiment 1

[0033] (1) Extraction of Genomic DNA of Cherry Blossom Varieties:

[0034] Take 0.01g of the young leaves of the cherry blossom variety to be tested, add liquid nitrogen and grind them thoroughly, and use the SDS-CTAB method to extract the genomic DNA. After multiple extractions, the crude genomic DNA extract of the cherry blossom variety is obtained. 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.

[0035](2) Design specific PCR amplification primers, the sequence of the primer pair is:

[0036] The upstream primer: 5′-AGCGGCCGCCGCTAGATGAG-3′ and the downstream primer: 5′-AGCGGCCGCCGAGACAAAGA-3′ were synthesized by Shanghai Bioengineering Te...

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Abstract

The invention relates to a pair of high-specificity molecular specific marker primers for sakura varieties including Plena Rosea, Sakuyahime and Ujou-shidare, and a method for quickly identifying the sakura varieties including Plena Rosea, Sakuyahime and Ujou-shidare. Primer sequences are as follows: an upstream primer: 5'-AGCGGCCGCCGCTAGATGAG-3'; a downstream primer: 5'-AGCGGCCGCCGAGACAAAGA-3'. The specific marker primers can be used for quick preliminary identification of the sakura varieties including Plena Rosea, Sakuyahime and Ujou-shidare, and the method is simple, fast and accurate and is a molecular means irreplaceable by apparent characteristic identification of the sakura varieties.

Description

(1) Technical field [0001] The present invention relates to molecular-specific marker primers for cherry blossom varieties "Yaehongzhidae", "Sakiye Hime" and "Yuqingzhidre", and use the primers to detect cherry blossom varieties "Yaehongzhidae", "Sakiyahime" and A method for rapid identification of "Rainy Branches". (2) Background technology [0002] Cherry blossoms belong to the genus Cerasus of the Rosaceae (Rosaceae) and are world-renowned ornamental plants. At present, there are more than 300 known varieties of cherry blossoms, which have great application value in gardens because of their beautiful plant shape, bright colors and long flowering period. In recent years, cherry blossoms have been planted on a large scale in various parts of my country and used in parks, schools, streets, courtyards and other green spaces, becoming one of the main ornamental flowers and trees in early spring. Beijing, Wuhan, Qingdao, Shanghai and other cities hold cherry blossom festivals...

Claims

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Application Information

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IPC IPC(8): C12Q1/68C12N15/11
CPCC12Q1/6895
Inventor 徐梁李海波杨少宗魏海龙赵庆杰柳新红
Owner ZHEJIANG FORESTRY ACAD
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