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Molecular marker primer composition for identifying inter-hybrid seeds of pears and apples and application thereof

A primer composition and molecular marker technology, applied in the field of molecular genetics and breeding, can solve the problems of less polymorphic sites, long waiting time, cumbersome and time-consuming operation, etc., and achieve improved accuracy and efficiency, stable and reliable results, and specificity. good effect

Active Publication Date: 2019-03-19
NANJING AGRICULTURAL UNIVERSITY
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AI Technical Summary

Problems solved by technology

Generally based on the method of morphological observation, identification is usually mainly based on the morphological characteristics of leaves, flowers, and fruits. Although the method is simple and intuitive, the identification process is subject to the subjective influence of the observer; at the same time, the observation index is often affected by the external environment. The influence of factors will also interfere with the identification results; while fruit trees usually have a childhood period of 5-8 years, it takes a long time to observe the fruit shape, and it is difficult to be identified in a short time
Using the chromosome identification method, species with different chromosome numbers can be easily identified. However, pears and apples belong to the Rosaceae family and have the same number of chromosomes (2n=34), so it is difficult to distinguish
The method of isoenzyme marking has been used to identify distant hybrid varieties, but its development is restricted by the lack of polymorphic sites and cumbersome and time-consuming operations.

Method used

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  • Molecular marker primer composition for identifying inter-hybrid seeds of pears and apples and application thereof
  • Molecular marker primer composition for identifying inter-hybrid seeds of pears and apples and application thereof
  • Molecular marker primer composition for identifying inter-hybrid seeds of pears and apples and application thereof

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Embodiment 1

[0027] Molecular markers for the identification of hybrids between pear and apple genera based on genome-specific sequences were obtained by the following methods:

[0028] a) Using 'Hongxing'×'Coomice', 'Jinguan'×'Dangshansuli', 'Fuji'×'Coomice' for intergeneric hybridization, 73, 37 and 201 plants were obtained respectively—a total of 311 F1 offspring of apple and pear plants.

[0029]b) The DNA extracted based on the CTAB method has a higher concentration and better quality, and the operation steps for extracting DNA are as follows:

[0030] (1) Collect the disease-free and clean young leaves of the parents and hybrid offspring in April in spring, store them in ziplock bags, and freeze them quickly in liquid nitrogen. Take 100mg leaves, add liquid nitrogen to the mortar and grind them thoroughly; then transfer the ground leaves to a 1.5ml centrifuge tube, add 400μl CTAB buffer, mix well and put them in a 65°C water bath for 30min, every Shake once every 10 minutes to full...

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Abstract

The invention discloses a molecular marker primer composition for identifying inter-hybrid seeds of pears and apples and an application thereof. Specific sequences of pear and apple genomes are detected by a method of whole genome comparison, seven pairs of molecular marker specific primers P13, P31, P33, A31, A33, A34 and A37 are developed newly, and positive and negative primer sequences are asshown in a table 1. By identifying the inter-hybrid offsprings of pears and apples by means of the specific primers, specific amplified strips of the pears, apples and hybrid offsprings can be obtained, separately, that is, the inter-hybrid seeds of the pears and the apples are detected truly. The molecular marker based on the specific sequences of the genomes can be used for identifying the inter-hybrid seeds of pears and apples and has important theoretical and practical guiding meaning in improving genetic breeding progress of fruit trees and accelerating molecular assisted selective breeding.

Description

technical field [0001] The invention belongs to the field of molecular genetic breeding, and provides a molecular marker primer composition for identifying hybrids between pear and apple genera and applications thereof. Background technique [0002] Apple (Malus Domestica Borkh) and pear (Pyrus L.) belong to the Rosaceae family and are important fruit tree species cultivated worldwide. At the same time, my country is the main origin of pears and apples, with a very long history of cultivation and rich genetic diversity , there are many local varieties with good quality, which are still the main planted varieties at present. However, along with the continuous improvement of economic development and people's living standards, the market needs more abundant and high-quality varieties of pears and apples to meet consumer demand. [0003] Distant hybridization, as a breeding method that breaks through the gene pool of the population and obtains excellent genetic material from oth...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6895C12N15/11C12Q1/6806
CPCC12Q1/6806C12Q1/6895C12Q2600/13C12Q2531/113
Inventor 吴俊李胜男张明月张绍铃齐开杰李甲明李晓龙薛蕾谢智华
Owner NANJING AGRICULTURAL UNIVERSITY
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