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A Molecular Detection Method for Rapid Identification of Plum/Apricot Plum Strains

An identification method, real plum technology, applied in the determination/inspection of microorganisms, chemical instruments and methods, botany equipment and methods, etc.

Active Publication Date: 2021-07-06
BEIJING FORESTRY UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In recent years, despite the completion of genome sequencing and genome resequencing in plum blossoms, there are still a large number of genes and their functions to be discovered, especially those related to traits of plum blossoms responding to seasonal climate change and related to plum blossom strains

Method used

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  • A Molecular Detection Method for Rapid Identification of Plum/Apricot Plum Strains
  • A Molecular Detection Method for Rapid Identification of Plum/Apricot Plum Strains
  • A Molecular Detection Method for Rapid Identification of Plum/Apricot Plum Strains

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0061] Example 1 Mining SNP molecular markers related to plum blossom stem color traits based on GWAS analysis

[0062] 1. Determination of phenotypic changes in plum blossom stem color

[0063] When plum blossoms respond to seasonal climate change, the color of the annual branches of Xingmei changes obviously along with the occurrence of leaf fall, while the annual branches of Zhenmei maintain the original green traits. Using the Royal Horticultural Society color chart (RHS) to measure the color of the annual branches of different strains of plum blossoms before and after defoliation. The results showed that there was no obvious difference in the color of the annual branches of different strains of plum blossoms before defoliation, but the differences in traits after defoliation were prominent. The annual stem color of apricot plum was dark purple (Royal Horticultural Society color card is 183-B), and the annual stem color of real plum was green ( The Royal Horticultural Soc...

Embodiment 2

[0067] Example 2 Cloning and analysis of PmEIL gene in GWAS-associated region

[0068] The total RNA extraction of plum blossom variety ('Jiangmei') was carried out according to the instructions of the plant total RNA extraction kit of Tiangen Biochemical Technology (Beijing) Co., Ltd.

[0069] The total RNA was reverse-transcribed into cDNA by the kit, and the ORF of the EIN3 gene was cloned by PCR using the cDNA as a template. The reaction system is template (cDNA) 100-150ng, PrimeSTAR HS (Premix) 25μL, upstream primer (EIL_F, SEQ ID NO.3) 0.3μM, downstream primer (EIL_R, SEQ ID NO.4) 0.3μM, ddH 2 O was added to 50 μL. Reaction conditions: 98°C for 2min; 98°C for 10s, 56°C for 15s, 72°C for 60s (35 cycles); 72°C for 5min; store at 4°C.

[0070] The target band was recovered by gel electrophoresis, ligated, transformed, and sequenced to obtain the target sequence. Combined with DNAMAN software to compare the sequence of EIN3 gene ORF, the matching degree was 98.87%. There...

Embodiment 3

[0072] Example 3 Analysis of PmEIL gene expression changes under simulated seasonal climate change conditions

[0073] 1. Treat plum blossoms with low temperature and short sunshine

[0074] 4-week-old real plums (‘Jiangmei’) and apricot plums (‘Yanxing’) with consistent growth were selected as test materials. First, adapt to the culture under the uniform condition of 24°C long-day light (16 / 8h light / dark) for 2 days, and then carry out 24°C long-day light (16 / 8h light / dark) and 24°C short-day light (8 / 16h light / dark) respectively , 6°C long-day light (16 / 8h light / dark) and 6°C long-day light (8 / 16h light / dark) conditions, cultivated continuously for 4 weeks, observed the growth of apricot plum and real plum, and compared the plant height and leaf number .

[0075] 2. Analysis of PmEIL gene expression

[0076] The total RNA of plum blossoms was extracted using the RNAsimple Total RNA Extraction Kit of Tiangen Biochemical Technology (Beijing) Co., Ltd., and the extraction me...

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Abstract

The invention relates to the technical fields of plant genetic engineering and molecular biology, in particular to a molecular detection method for quickly identifying true plum / apricot plum strains. The present invention provides plum blossom PmEIL protein, the sequence of which is shown in SEQ ID NO.1. PmEIL protein is involved in the response of plum blossoms to seasonal climate changes. There are obvious differences in the regulation methods of real plum and apricot plum strains by seasonal climate changes such as temperature and sunshine. Quickly and accurately identify plum blossom strains, shorten the breeding cycle, and greatly reduce the breeding workload. The invention also provides a SNP molecular marker related to the PmEIL gene and related to plum blossom stem color, which provides a basis and a molecular tool for the identification of plum blossom stem color traits and the identification of true plum / apricot plum strains.

Description

technical field [0001] The invention relates to the technical field of plant genetic engineering and molecular biology, in particular to plum blossom ethylene-insensitive protein PmEIL, SNP molecular markers related to plum blossom ethylene-insensitive protein PmEIL and plum blossom stem color, and the use of ethylene-insensitive protein PmEIL and plum blossom Molecular detection method for rapid identification of real plum / xingmei strains with stem color-related SNP molecular markers. Background technique [0002] Plum blossom (Prunus mume Sieb.et Zucc.) is a plant of the genus Prunus (Rosaceae) in the family Rosaceae. It is one of the ten traditional famous flowers in China. It has important ornamental and economic value. It has a history of more than 3,000 years of introduction and cultivation (Chen Junyu. Illustrated Records of Chinese Plum Blossom Varieties [M]. China Forestry Press, 2010). Plum blossoms are divided into three major systems, namely the Zhenmei system (...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07K14/415C12N15/29C12N15/11C12Q1/6895
CPCC07K14/415C12Q1/6895C12Q2600/13C12Q2600/156
Inventor 郑唐春张启翔李平李璐璐程堂仁王佳袁存权
Owner BEIJING FORESTRY UNIVERSITY
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