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A method for identifying abortion-related proteins in chrysanthemum distant hybrid embryos

A technology of distant hybridization and related proteins, applied in measuring devices, instruments, scientific instruments, etc., can solve the problem of difficult to find and control embryo-related genes, and achieve high repeatability.

Inactive Publication Date: 2015-09-30
NANJING AGRICULTURAL UNIVERSITY
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Problems solved by technology

There are usually thousands of genes that control the development of plant embryos, and it is difficult to find the relevant genes that control embryo abortion directly from the genetic level

Method used

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  • A method for identifying abortion-related proteins in chrysanthemum distant hybrid embryos

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Embodiment Construction

[0025] The present invention will be described in further detail below in conjunction with specific embodiments.

[0026] The method provided by the present invention utilizes iTRAQ technology to quickly identify differential proteins between Chrysanthemum and closely related genera after distant hybridization and pollination of normally developed embryos and aborted embryos. The specific implementation examples are as follows:

[0027] 1. Distant hybridization and sampling

[0028] 1.1 Distant hybridization

[0029] Using the cultivated chrysanthemum 'Yuhua Luoying' (2n=6x=54) as the female parent, select well-developed flower buds, and emascate when the tongue-shaped flowers are just exposed, that is, remove all the bisexual tubular florets in the inner round, and cut off the tongue-shaped flowers at the same time. Flower petals until stigmas are visible (do not hurt stigmas), and bag strictly. When the stigma of the female parent protrudes and splits to present acute angl...

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Abstract

The invention discloses a method for identifying embryo abortion associated proteins of distant hybridization of chrysanthemum, and belongs to the field of chrysanthemum breeding and proteomics. The method comprises the steps of planting the small chrysanthemum as a female parent, taking wild chrysanthemum nankingense as a male parent for distant hybridization, cutting off pollinated inflorescences at different times after pollination, sampling ovaries with complete and plump morphological development and ovaries with abnormal and hollow morphological development respectively, immediately quick-freezing in liquid nitrogen after sampling, storing in a refrigerator at -80 DEG C, extracting proteins from samples by a TCA (Trichloroacetic Acid) / acetone precipitation method, quickly identifying differential proteins by an iTRAQ (Isobaric Tags for Relative and Absolute Quantitation) and mass-spectrometric technique, performing bioinformatic analysis, and obtaining the embryo abortion associated protein in the distant hybridization breeding of the chrysanthemum. The method can quickly and accurately identify the embryo differential proteins with different morphologies in different developmental stages during embryonic development of the chrysanthemum for the chrysanthemum and affinis wild species of the chrysanthemum, and a research foundation is provided for solving embryo abortion of the distant hybridization of the chrysanthemum.

Description

technical field [0001] The present invention relates to the fields of chrysanthemum hybrid breeding and proteomics, and relates to a rapid detection and identification method for the differential proteins of normal and aborted embryos in different developmental stages of embryos after chrysanthemum distant hybridization, in particular to a method for identifying aborted embryos of chrysanthemum distant hybridization Methods for educating related proteins. Background technique [0002] Chrysanthemum originates in China, and is one of my country's top ten traditional famous flowers, the world's four major cut flowers and top ten potted flowers, and occupies a very important position in flower production. At present, there are thousands of chrysanthemum varieties in China, but most of the varieties with excellent ornamental properties have poor resistance, while the wild resources of most closely related species contain excellent characteristics that many cultivated chrysanthem...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N30/89
Inventor 滕年军张凤姣黄至喆陈发棣房伟民蒋甲福管志勇陈素梅刘兆磊
Owner NANJING AGRICULTURAL UNIVERSITY
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