Molecule mark identification method for plum pollen autocopulation non-compatible genotype

Abstract

The invention discloses a molecular marking identifying method of autocopulation uncongenial gene type (S gene type) of plum in the biological technical domain, which is characterized by the following: adopting marked primer to augument different kinds of plum DNA; obtaining the augumented segment with 1300bp; marking pollen SFB gene; using MboI enzyme augument product to produce enzyme-cutting band; displaying different enzyme-cutting bands according to different unit typed pollen SFB gene; affirming autocopulation uncongenial gene type; setting the same band as the same S gene type and vice versa.

Description

1. Technical field

[0001] The invention provides a molecular marker method for identification of plum pollen self-incompatibility genotype (S genotype for short), belongs to the field of biotechnology, and is used for identification of fruit tree self-incompatibility genotype varieties and selection of breeding parents and the configuration of its pollinating varieties. 2. Background technology

[0002] The plum fruit is sweet in flavor and rich in nutrition. The fruit contains a variety of vitamins, minerals, organic acids, etc., and is a fresh fruit with high nutritional value. Prunus belongs to the gametophytic self-incompatibility fruit trees. Most varieties are self-fertile or have a very low rate of self-fertilization. In production, it is necessary to configure suitable pollination trees to ensure fruit setting. In addition, there is also the phenomenon of hybrid incompatibility between varieties, so there is a certain blindness in the allocation of pollinated variet...

Examples

Embodiment Construction

[0024] 1. Refer to the method of Doyle and Doyle (Doyle JI, DoyleJL. A rapid DNA isolation procedure for small quantities of fresh leaf tissue. Phytochem Bull, 1987, 19: 11-15) to extract the genomic DNA of 11 plum varieties: banana plum, royal Diamond, 82-1-109, Santa rosa ShSk, Younai, Wixon, Rose Plum, Nvgelei, Huangpi Plum, Black Jewel, Gaixian Dali, all varieties come from the National Germplasm Bank of Xingli (Liaoning Xiongyue)".

[0025] 2. By comparing the conserved regions of cherry, plum, almond and apricot pollen SFB genes, design general primers for plum pollen SFB gene according to the conserved sequence:

[0026] Forward primer 5′-TTKSCHATTRYCAACCKCAAAAG-3′

[0027] Reverse primer 5′-CAMRAATTCGATTTCGYCATATTTC-3′

[0028] The plum genomic DNA was amplified with the designed universal primers.

[0029] Reaction system: 25μL total reaction volume, 10×PCR Buffer 2.5μL 3.0mM MgCl 2 , 0.2mM dNTP, each primer 0.1μM, 20-50ng template, Taq enzyme 1.0U.

[0030] The ...

1. Technical field

[0001] The invention provides a molecular marker method for identification of plum pollen self-incompatibility genotype (S genotype for short), belongs to the field of biotechnology, and is used for identification of fruit tree self-incompatibility genotype varieties and selection of breeding parents and the configuration of its pollinating varieties. 2. Background technology

[0002] The plum fruit is sweet in flavor and rich in nutrition. The fruit contains a variety of vitamins, minerals, organic acids, etc., and is a fresh fruit with high nutritional value. Prunus belongs to the gametophytic self-incompatibility fruit trees. Most varieties are self-fertile or have a very low rate of self-fertilization. In production, it is necessary to configure suitable pollination trees to ensure fruit setting. In addition, there is also the phenomenon of hybrid incompatibility between varieties, so there is a certain blindness in the allocation of pollinated variet...

Examples

Embodiment Construction

[0024] 1. Refer to the method of Doyle and Doyle (Doyle JI, DoyleJL. A rapid DNA isolation procedure for small quantities of fresh leaf tissue. Phytochem Bull, 1987, 19: 11-15) to extract the genomic DNA of 11 plum varieties: banana plum, royal Diamond, 82-1-109, Santa rosa ShSk, Younai, Wixon, Rose Plum, Nvgelei, Huangpi Plum, Black Jewel, Gaixian Dali, all varieties come from the National Germplasm Bank of Xingli (Liaoning Xiongyue)".

[0025] 2. By comparing the conserved regions of cherry, plum, almond and apricot pollen SFB genes, design general primers for plum pollen SFB gene according to the conserved sequence:

[0026] Forward primer 5′-TTKSCHATTRYCAACCKCAAAAG-3′

[0027] Reverse primer 5′-CAMRAATTCGATTTCGYCATATTTC-3′

[0028] The plum genomic DNA was amplified with the designed universal primers.

[0029] Reaction system: 25μL total reaction volume, 10×PCR Buffer 2.5μL 3.0mM MgCl 2 , 0.2mM dNTP, each primer 0.1μM, 20-50ng template, Taq enzyme 1.0U.

[0030] The ...

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