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Specific labeling primers for Sakura Yae, Sakuya Hime and Yuqing Zhida

A technology of eightfold red branch hanging and rain branch hanging, applied in the direction of recombinant DNA technology, DNA / RNA fragments, etc., can solve the problems of difficult identification of phenotypic characteristics, application and promotion of garden plants, and difficulty in breeding new varieties, etc., to achieve the method simple effect

Active Publication Date: 2017-09-22
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 (Cerasusserrulata'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'), Yae Red Branch Weeping (C.spachiana'Plena Rosea') , Ichihara Huwei (C.serrulata'Albo Plena'), Saki Yeji (Cerasus×yedoensis'Sakuyahime'), Suzaku (C.serrulata'Shujaku'), Yang Guifei (C.serrulata'Mollis'), Yuqing Zhiwei ( Application of C.spachiana'Ujou‐shidare'), Chrysanthemum weeping cherry (C.serrulata'Plena-pendula'), Hirano sister back (C.serrulata'Imose'), Asahiyama cherry (C.serrulata'Asahiyama') in gardens It is extremely widespread, 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 from the aspect of phenotypic characteristics, which brings many difficulties to the application, promotion and selection of new varieties of garden plants. 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 labeling primers for Sakura Yae, Sakuya Hime and Yuqing Zhida
  • Specific labeling primers for Sakura Yae, Sakuya Hime and Yuqing Zhida
  • Specific labeling primers for Sakura Yae, Sakuya Hime and Yuqing Zhida

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Embodiment 1

[0033] (1) Extraction of genomic DNA of cherry blossom varieties:

[0034] Take 0.01 g of young leaves of the cherry cultivar to be tested, add liquid nitrogen to grind thoroughly, and use the SDS-CTAB method to extract the genomic DNA. The crude DNA extract was purified by Magabio nucleic acid purification kit (Bioer, Hangzhou, China), and then 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 until use.

[0035](2) Design specific PCR amplification primers, and the sequences of the primer pairs are:

[0036] Upstream primer: 5'-AGCGGCCGCCGCTAGATGAG-3' and downstream primer: 5'-AGCGGCCGCCGAGACAAAGA-3', synthesized by Shanghai Bioengineering Technology Co., Ltd.

[0037] (4) PCR amplification:

[0038] Composition of PCR reaction solution: 10×PCR Buffer...

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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 "Yaehongzhitu", "Sakuyeji" and "Yuqingzhituo", and the use of the primers for the sakura varieties "Yaehongzhitu", "Sakuyeji" and "Sakuyeji". A method for rapid identification of "rainy branches". (2) Background technology [0002] Cherry blossoms belong to the genus Cerasus of the Rosaceae family and are world-famous ornamental plants. At present, there are more than 300 varieties of cherry blossoms, which have great application value in gardens because of their beautiful plant shape, bright flower color and long duration of the entire flowering period. In recent years, cherry blossoms have been planted on a large scale throughout 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...

Claims

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Application Information

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Patent Type & Authority Patents(China)
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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