Analysis method for researching formation of variegation of nelumbo nucifera based on combination of transcriptomics with proteomics TMT

A technology that combines proteins and analysis methods, applied in the application field of new technologies in hydropower engineering, can solve problems such as unreported invention patents, and achieve the effects of wide application range, accurate quantification, and fast analysis speed

Pending Publication Date: 2019-10-15
CHINA THREE GORGES CORPORATION
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Problems solved by technology

Proteomics is mostly used in medical research, and it is used in the study of plant mottling formation. After searc

Method used

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  • Analysis method for researching formation of variegation of nelumbo nucifera based on combination of transcriptomics with proteomics TMT
  • Analysis method for researching formation of variegation of nelumbo nucifera based on combination of transcriptomics with proteomics TMT
  • Analysis method for researching formation of variegation of nelumbo nucifera based on combination of transcriptomics with proteomics TMT

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Embodiment 1

[0072] 1. Collect the flower buds on the first day of water emergence (the uncolored petals are denoted as S_1) and the flower buds on the 15th day of water emergence (the white area of ​​the petals is S2_1 and the coloring spot area of ​​​​the petals is S2_2), and the fresh petals are collected and quickly frozen with liquid nitrogen , stored at -80°C.

[0073] 2. Extraction of RNA. Because the petals of lotus contain more polysaccharides and polyphenols, the RNA in the petals of lotus ‘Dasajin’ was rapidly extracted using the Magen kit RNA.

[0074] 3. Build the library. After the sample is qualified, the magnetic beads with Oligo(dT) are used to enrich the mRNA of eukaryotes by combining A-T complementary pairing with the polyA tail of the mRNA. Then fragmentation buffer was added to break the mRNA into short fragments, using the mRNA as a template, a strand of cDNA was synthesized with six base random primers (random hexamers), and then a second strand of cDNA was synthe...

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Abstract

The invention relates to an analysis method for researching formation of variegation of nelumbo nucifera based on combination of transcriptomics with proteomics TMT, and provides a new method for disclosing a molecular mechanism of the formation of the variegation of plants. Samples of "Dasajin" nelumbo nucifera petals are collected, through transcriptome Illumina HiSeq sequencing, expression difference genes on an anthocyan metabolic pathway are screened, besides, proteins in the samples are extracted, the concentration of the proteins is determined, the quality is detected, through TMT labelmarking and 2D-LC-MS analysis, bioinformatics analysis is performed through combining with the proteomics, the proteins for adjusting and controlling expressing level differences on the anthocyan metabolic pathway are screened, the genes having differences in the transcriptome are related into the proteomics, and key proteins and corresponding genes for adjusting and controlling the variegation of "Dasajin" nelumbo nucifera are screened, so that the purpose of understanding a manner of the proteins and the genes for adjusting the formation of the variegation of the "Dasajin" nelumbo nuciferais achieved, and a new thinking and a new method are provided for the formation mechanism of the variegation of other plants.

Description

technical field [0001] The invention relates to the field of application of new technologies in hydropower engineering, in particular to a method for analyzing the formation of lotus spots based on transcriptomics combined with proteomics TMT. Background technique [0002] Lotus (Nelumbo nucifera) belongs to Nelumbonaceae Nelumbo. As an ancient perennial aquatic herbaceous flower, it has high ornamental value, and flower color is one of its important ornamental elements, which determines the ornamental value of lotus. . However, there are few studies on the formation mechanism of lotus flower spots, so it is imperative to use omics methods to study the formation mechanism of lotus flower spots, which will open up a new research path for the study of plant flower spot formation mechanisms. With the completion and continuous development of science and technology, it is not possible to comprehensively, effectively and accurately study the physiological metabolism of plant vari...

Claims

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Application Information

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IPC IPC(8): C12Q1/6895G01N33/68G16B20/00G16B30/00G16B40/00
CPCC12Q1/6895G01N33/6848G16B20/00G16B30/00G16B40/00C12Q2600/158C12Q2600/13G01N2333/415
Inventor 李林宝赵凯歌张国禹黄桂云吴笛吴锦华杨兰芳马晓波陈会员镇巧玲邱利文张海波张俊胡梅香张定军汪磊望雄英李翩翩
Owner CHINA THREE GORGES CORPORATION
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