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Method for predicting wheat heterosis by mitochondrial proteome technology

A proteome and technology prediction technology, applied in the field of agricultural bioengineering, can solve problems such as inability to predict, and achieve the effect of comprehensive species and information

Inactive Publication Date: 2013-08-28
TIANJIN NORMAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The above methods are very helpful for us to understand the formation mechanism of heterosis, but it is not yet possible to predict

Method used

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  • Method for predicting wheat heterosis by mitochondrial proteome technology
  • Method for predicting wheat heterosis by mitochondrial proteome technology
  • Method for predicting wheat heterosis by mitochondrial proteome technology

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0041] (1) Obtaining the hybrid F1: the female parent of the cultivated wheat Jing 411 with excellent agronomic traits (gifted by the School of Agricultural Biotechnology, China Agricultural University), and the translocation line Ta-6DL / 6VS resistant to powdery mildew wheat as the male parent (Nanjing Agricultural University donated by the Institute of Agricultural Biotechnology) to obtain hybrid offspring F1: Jing 411 (♀)×Ta-6DL / 6VS (♂); the detailed process of hybridization includes flowering adjustment, ear selection, whole ear, emasculation, harvesting pollination, pollination and harvesting.

[0042] Adjust flowering period: Jing 411 is the female parent, sow in stages every 5 days, the wheat translocation line Ta-6DL / 6VS is the male parent and Jing 411 are sown at the same time, and the plants that meet the flowering stage are crossed.

[0043] Ear selection: before emasculation of the female parent, select a well-developed, robust main stem ear with the typical charact...

Embodiment 2

[0052] Methods for the detection of mitochondrial proteins.

[0053](1) Two-dimensional electrophoresis method: the first isopoint focusing electrophoresis (IEF): 30% acrylamide and methylene bisacrylamide gel, containing 10% APS, 2% NP40, 8 mol / L urea, ampholyte stock solution pH (3-10) and pH (5-7); the upper channel buffer of IEF gel is 0.02mol / L NaOH, and the lower channel buffer is 0.01 mol / L H 3 PO 4 solution; power on, 200 V×15 min, 300 V×30 min, 400 V×60 min for pre-electrophoresis; Rinse with double distilled water 2-3 times. Pipette 40 microliters of the protein to be tested into the sample, gently cover the sample with NaOH buffer until the tube is full, fill the upper tank with 0.02mol / L NaOH buffer, run electrophoresis at 400 V for 14 hours, and electrophoresis at 800 V for 1 hour. After stopping the electrophoresis, peel off the gel strip, wash it with deionized water for 1-2 times, and quickly put it into the equilibrium solution (2% SDS, 10% glycerol, 5% β-m...

Embodiment 3

[0056] Method for predicting heterosis in wheat: cut the mitochondrial protein target spot from the gel, grind it into powder and wash it with water; wash the gel with a mixture of 50 mM ammonium bicarbonate and acetonitrile (volume ratio 1:1) for 15 minutes to fade, Then immerse in 100% acetonitrile for dehydration for 10 minutes, discard the acetonitrile, and dry the sample in a vacuum desiccator for 15 minutes; place the dried gel powder in 25mM ammonium bicarbonate solution containing 50ng trypsin to swell and incubate at 37°C overnight; extract the gel with 50% acetonitrile, 5% trifluoroacetic acid to collect the peptide mixture; mitochondrial protein samples are mixed with α-cyano-3-hydroxycinnamic acid saturated with 50% acetonitrile, 0.1% trifluoroacetic acid in equal volumes , placed on the sample target; peptide mass was detected by ABI4700 instrument, and the mass accuracy was between 10-50ppm; peptide mass fingerprint (PMF) was searched in the NCBI database with MAS...

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Abstract

The invention discloses a method for predicting wheat heterosis by a mitochondrial proteome technology. The invention mainly relates to cultivation of wheat Jing 411, wheat translocation line Ta-6DL / 6VS and its hybridization progeny F1 (Jing 411(female)*Ta-6DL / 6VS(male)). By separating the mitochondrial proteome of the material, and combining a MALDITOFMS technology by utilizing bidirectional gel electrophoresis, obvious difference showing in leaf mitochondrial proteome between wheat hybrid and amphiphilic is discovered, and the difference proteins has four kinds of expression modes: an additional type (ADT), a prejudiced amphiphilic type (PDT), a higher than amphiphilic type (HPDT) and a lower than amphiphilic type (LPDT), wherein expression frequencies of the four modes are respectively 15.2%, 54.5%, 24.2% and 6.1%. Wherein, spot 22 (alcohol dehydrogenase) continuously shows higher than amphiphilic type expression in different growth periods of the wheat hybrid F1 and amphiphilic. The protein can be used for advantage prediction of the hybrid F1.

Description

[0001] Statement on funding research or development: [0002] This invention is carried out under the support of the National Natural Science Foundation of China (31071671) and the major support project of Tianjin Science and Technology Commission (10ZCKFNC00200). technical field [0003] The invention belongs to the technical field of agricultural bioengineering, and relates to a technique for predicting wheat heterosis dynamic mitochondrial proteome and its application. Background technique [0004] Heterosis refers to the phenomenon that the offspring of crops exceed their parents in vigor, growth vigor and yield (Song et al, 2009). Because of its huge commercial value, it has always been highly valued by academic circles and production units. The successful utilization of rice heterosis in my country has reduced the rice cultivation area from 36.5 Mha in 1975 to 30.5 Mha in 2000, but the output has increased from 12.8 billion tons to 18.9 billion tons, that is, from 3....

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A01H1/02G01N27/447G01N27/62
Inventor 彭永康范宝莉刘晓颖王振英陈宏
Owner TIANJIN NORMAL UNIVERSITY
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