Primers for identifying sex of Diospyros kaki and application of primers

A persimmon tree sex technology, which is applied to primers for identifying the sex of persimmon trees and its application field, can solve problems such as non-gender identification, and achieve the effect of high sensitivity and simple method

Pending Publication Date: 2020-12-11
RES INST OF NON TIMBER FORESTRY CHINESE ACAD OF FORESTRY
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

It is worth noting that due to the limitation of experimental materials, this study did not conduct sex identification on the complete male plant of persimmon germplasm resources unique to my country

Method used

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  • Primers for identifying sex of Diospyros kaki and application of primers
  • Primers for identifying sex of Diospyros kaki and application of primers
  • Primers for identifying sex of Diospyros kaki and application of primers

Examples

Experimental program
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Effect test

Embodiment 1

[0040] Example 1 Molecular marker primer MF

[0041] Sangon Bioengineering (Shanghai) Co., Ltd. was commissioned to synthesize the molecular marker primer MF for identifying the sex of persimmon trees. The primer information is shown in Table 1.

[0042] Table 1

[0043]

Embodiment 2

[0045] Utilize the primer MF of distinguishing persimmon tree sex in the embodiment to carry out the application of distinguishing persimmon tree sex:

[0046] (1) Extract the DNA of persimmon leaves;

[0047] The genomic DNA of 86 persimmon trees (36 complete female plants, 33 complete male plants, and 17 monoecious plants) were extracted with the Ezup column type plant genomic DNA extraction kit (Shenggong, China). The specific methods are as follows:

[0048] a) Weigh 0.5g of fresh plant leaves, fully grind them in liquid nitrogen, and transfer them to a sterilized 1.5ml centrifuge tube;

[0049] b) Add 700ul of Buffer PCD preheated at 65°C and 14ul of β-mercaptoethanol to the above centrifuge tube, mix well, bathe in water at 65°C for 20min, and shake occasionally for 2-3 times;

[0050] c) Add an equal volume of chloroform to the mixture, mix well, and centrifuge at 12000rpm for 5min;

[0051] d) Pipette the supernatant into a new 1.5ml centrifuge tube, add the same vol...

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PUM

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Abstract

The invention discloses primers for identifying the sex of Diospyros kaki. The primers comprise an MF forward primer and an MF reverse primer and are shown as SEQ ID No. 1 and SEQ ID No. 2 in a sequence table. The application of the primers for identifying the sex of the Diospyros kaki comprises the following steps of extracting Diospyros kaki genome DNA; carrying out PCR amplification on the extracted DNA by utilizing the forward primer and the reverse primer; carrying out clone sequencing on a PCR product; and performing sex determination on the Diospyros kaki according to a sequencing result. The MF primer provided by the invention can be used for predicting male and female types of persimmon germplasm resources, whether male flowers blossom or not can be identified by detecting whethera specific nucleotide sequence exists in a sample or not under the condition that the gender source of the sample is unknown, the accuracy of an identification result is greater than 91%, and a simple, convenient, quick and effective molecular marker is provided for identifying the sex of Diospyros kaki.

Description

technical field [0001] The invention relates to the technical field of molecular identification. Specifically, it is a primer for identifying the sex of persimmon trees and its application. Background technique [0002] Persimmon (Diospyros kaki Thunb.) belongs to the persimmon family (Ebenaceae) of the genus Diospyros. It is an important woody food tree species native to my country. are getting more and more attention. Most cultivars of persimmon are completely female plants (only female flowers are opened), and a few are monoecious plants (female flowers and male flowers are opened at the same time). In addition, there are a small number of completely male plants (only male flowers) in the wild mountainous areas of Jiangsu, Hunan, Hubei, Jiangxi, Guangxi and other places in my country. [0003] Japanese scholars used monoecious and completely female persimmon trees as research materials, and analyzed their sex-determining loci, and found that the production of male flowe...

Claims

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Application Information

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IPC IPC(8): C12Q1/6879C12Q1/6895C12Q1/686C12N15/11
CPCC12Q1/6879C12Q1/6895C12Q1/686C12Q2565/125
Inventor 孙鹏傅建敏王丽媛李华威索玉静韩卫娟刁松锋王艺儒
Owner RES INST OF NON TIMBER FORESTRY CHINESE ACAD OF FORESTRY
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