Molecular-specific marker primers and detection methods for cherry blossom cultivar "Sakiyahime"
A marker primer and specific technology, applied in the direction of DNA/RNA fragment, recombinant DNA technology, etc., can solve the problems of difficult identification of phenotypic characteristics, difficulty in application and promotion of garden plants, and difficulty in breeding of new varieties, and achieve the effect of simple method.
Active Publication Date: 2017-08-25
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
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Embodiment 1
[0040] (1) Extraction of Genomic DNA of Cherry Blossom Varieties:
[0041] 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.
[0042] (2) Design specific PCR amplification primers, the sequence of the primer pair is:
[0043] The upstream primer: 5′-TGAGTCCAAACCGGACCGAGAAG-3′ and the downstream primer: 5′-GACTGCGTACGAATTGACGGGAAT-3′ were synthesized by Shanghai Bioengineer...
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The invention relates to a molecular specific marker primer with high specificity for Sakura variety 'Cerasus yedoensis Sakuyahime' and a method capable of rapidly identifying the Sakura variety 'Cerasus yedoensis Sakuyahime'. The primer has the following sequences: an upstream primer: 5'-TGAGTCCAAACCGGACCGAGAAG-3', and a downstream primer: 5'-GACTGCGTACGAATTGACGGGAAT-3'. The molecular specific marker primer disclosed by the invention can realize rapid preliminary identification of the Sakura variety 'Cerasus yedoensis Sakuyahime', and the method is simple, rapid and accurate and is a molecular method which cannot be replaced for identifying apparent characteristics of the Sakura variety.
Description
[0001] (1) Technical field [0002] The invention relates to a molecular-specific marker primer of the cherry blossom variety "Sakiya Hime" and a method for rapidly identifying the cherry blossom variety "Sakiya Hime" by using the molecular-specific marker primer. [0003] (2) Background technology [0004] 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 every year, and the cherry blossom industry has...
Claims
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IPC IPC(8): C12Q1/68C12N15/11
CPCC12Q1/6895
Inventor 柳新红徐梁李海波王青华赵庆杰郭佳余建国
Owner ZHEJIANG FORESTRY ACAD