Method for identifying litchi varieties by using wide targeted metabonomics technology
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A metabolomics and variety technology, applied in the field of litchi variety identification, can solve the problems of low efficiency, hindering substantial breakthroughs in molecular marker-assisted selection breeding technology, and poor reliability.
Pending Publication Date: 2020-09-29
POMOLOGY RES INST GUANGDONG ACADEMY OF AGRI SCI
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[0004] However, because most of these germplasm resources have unclear genetic relationships or sources, the phenomenon of mixed varieties and substandard ones is serious. In order to improve breeding efficiency, discover and utilize excellent germplasm, and protect the interests of consumers to the greatest extent, it is necessary to find a An effective tool for litchi germplasm identification, providing more effective development means for the development of litchi industry
[0005] At present, observation of morphological characteristics and traits is still the main method for identification of litchi varieties. Practice has shown that this method has low efficiency and poor reliabi...
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Embodiment 1
[0042] Taking the Identification of "Guiwei" Litchi as an Example
[0043] 1. Mass spectrometric identification of metabolites in litchi variety library, construction of peak area database, screening of differential metabolites and construction of heatmap.
[0044] (1) Preparation of lychee samples, the process is as follows: 10 known conventional commercial varieties ("Baitangying", "Feizixiao", "Feicui", "Guiwei", "Jinbaoyin", "Nuomici" ", "Sanyuehong", "Xianjinfeng", "Yujinqiu", respectively referred to as BTY, FZX, FC, GW, JBY, NMC, SYH, XJF, YJQ in this application) litchi samples in the mature period ( Maturity is 100%) after picking, the pulp is cleaned, stripped, vacuum freeze-dried, and ground to powder with a grinder; 100 mg of powder is accurately weighed and dissolved in 1.0 mL of 70% (v / v) methanol extract; The dissolved samples were vortexed rapidly 3 times, 5min each time, sonicated once (350W, 35kHz), and refrigerated overnight at 4°C; after refrigerated centr...
Embodiment 2
[0055] Take "Yujinqiu" litchi as an example
[0056] 1. Mass spectrometric identification of metabolites in litchi variety library, construction of peak area database, screening of differential metabolites and construction of heatmap.
[0057] (1) The preparation of lychee sample, process is: after the lychee sample of known variety name (same as embodiment 1) is plucked at the full maturity stage (maturity is 90%), the pulp is cleaned and peeled off to get the pulp, and then vacuum freeze-dried, Use a grinder to grind to powder; accurately weigh 100mg of powder, and dissolve it in 1.0mL70% (v / v,) methanol extract; the dissolved sample is vortexed rapidly 3 times, 5min / time, ultrasonicated 1 time (350W , 35kHz), 4°C refrigerator overnight; after refrigerated centrifugation (rotational speed 10,000g, 10min), absorb the supernatant, filter the sample with a 0.22μm microporous membrane and put it into a sample vial for LC-MS / MS analysis . At least 3 biological replicates were p...
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Abstract
The invention discloses a method for identifying litchi varieties by using a wide targeted metabonomics technology. The method comprises the following steps: (1) obtaining a known litchi variety endogenous small molecule metabolite peak area database through litchi sample preparation, litchi sample detection and analysis, litchi sample qualitative and quantitative analysis and metabonomics data processing and analysis, screening to obtain 69 common difference significant metabolites, and generating a heatmap; and (2) obtaining the peak area of the endogenous small molecule metabolite of the unknown variety, combining the peak area with the peak area database of the known variety, carrying out multivariate statistical analysis, carrying out clustering analysis on the screened common differential metabolite, and judging the known variety which is firstly gathered with the variety to be identified as the variety to be identified or the similar variety. According to the method, the endogenous small molecule metabolites are obtained by utilizing a wide targeted metabonomics analysis technology, common difference significance metabolites of litchis are obtained through multivariate statistical analysis, and a visual map is generated and can be used for litchi variety identification.
Description
technical field [0001] The invention belongs to the technical field of litchi variety identification, and in particular relates to a method for identifying litchi varieties by using extensive targeted metabolomics technology. [0002] technical background [0003] Litchi (Litchi chinensis Sonn.) is a typical subtropical evergreen fruit tree. It has been cultivated in China for more than 2,000 years. In 2018, the cultivated area of litchi in my country was 552,000 ha 2 , the output reached 2.297 million tons, and the cultivation area and output ranked first in the world. After long-term natural selection and artificial cultivation, my country's litchi has formed a large number of ecological types and local varieties, and the germplasm resources are extremely rich. Up to now, there are more than 500 varieties, and there are more than 30 varieties sold on a commercial scale. , which is a unique advantage of our country. [0004] However, because most of these germplasm resour...
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