DNA (deoxyribonucleic acid) fragment, primers and method for identifying gender of salak

A technology of snake skin fruit and identification method, which is applied in the field of molecular biology and can solve the problems of difficulty in obtaining steps and process data, inability to identify snake skin fruit gender and the like.

Active Publication Date: 2012-05-02
RES INST OF TROPICAL FORESTRY CHINESE ACAD OF FORESTRY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In 2009, it was reported in Indonesia that isoenzymes could be used to identify the sex of the snake skin fruit seedlings, but the specific steps and process data of this technology were difficult to obtain, so the sex of the snake skin fruit seedlings could not be identified in domestic production. The consequences of seedling afforestation are still unavoidable

Method used

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  • DNA (deoxyribonucleic acid) fragment, primers and method for identifying gender of salak
  • DNA (deoxyribonucleic acid) fragment, primers and method for identifying gender of salak
  • DNA (deoxyribonucleic acid) fragment, primers and method for identifying gender of salak

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Embodiment 1

[0015] Genomic DNA was extracted from 10 male and female snake skin fruit strains whose sex had been determined, followed by RAPD amplification and electrophoresis according to the following method.

[0016] Step 1: Leaf Collection

[0017] Take the upright and unexpanded leaves of the snake skin fruit as the first leaf, and the leaves of the third leaf counted down as the collected leaves, take a 5 cm × 5 cm leaf on the leaf, wipe off the dust and water on the surface of the leaf sample, and Place in a plastic bag, then seal the plastic bag and place in an ice pack.

[0018] Step 2: Genomic DNA Extraction

[0019] Add 4 ml of 2% CTAB extract (CTAB20 g / L, NaCl 81.816 g / L, EDTA9.306 g / L, Tris-HCl 12.114 g / L, pH8.0) into a 10 ml centrifuge tube, then add 2% β-mercaptoethanol and 5% PVP-40, incubated at 65°C. Weigh 1.2 grams of the leaf sample of the snake skin fruit, cut the leaf sample into pieces, put it into a mortar, add liquid nitrogen and grind it to powder. Add the gr...

Embodiment 2

[0027]

[0028]

[0029]

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Abstract

The invention discloses a DNA (deoxyribonucleic acid) fragment, primers and a method for identifying gender of salak. The DNA specific fragment for identifying the gender of salak has a nucleotide sequence as shown in SEQ ID NO.1. Identifying primers are as shown in SEQ ID NO.2 and SEQ ID NO.3. The identifying method comprises the steps of: the identifying primers are taken as amplification primers, the genome DNA of a sample serves as a template to carry out PCR (polymerase chain reaction), the PCR product is subjected to electrophoresis detection, the sample with a fragment of 1579bp band, occurring in the electrophoresis results, is a male plant, while the sample without a band at the same part is a female plant. The primers and method disclosed by the invention can accurately identifythe gender, i.e. male and female of salak. Compared with the traditional method for identifying the gender of salak plant by naked eyes, the method disclosed by the invention has the advantages of being simple and convenient and being capable of identifying anytime, thus creating conditions for effectively and reasonably planting salak.

Description

Technical field: [0001] The invention belongs to the field of molecular biology, and in particular relates to a DNA fragment, a primer and a method for identifying the sex of snake skin fruit. Background technique: [0002] Snake skin fruit (Salacca zalacca) is a tropical fruit tree with unique fruit shape, rich in calcium and iron, convenient fruit picking, and suitable for cultivation in Hainan and Yunnan of my country. Sexual reproduction is an important way for the reproduction of Snakeskin fruit, but Snakeskin fruit is a dioecious plant, and the sex of the seedlings produced by sexual reproduction is difficult to identify with naked eyes. Afforestation of unidentified seedlings will easily lead to uneven distribution of female plants, high restoration costs and low yield per unit area. The traditional visual identification of the sex of the snake skin fruit plant needs to wait for the plant to start to reproduce and grow, but the age of the first reproductive growth of...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68C12N15/11
Inventor 尹天光李荣生李发根杨锦昌邹文涛李建光
Owner RES INST OF TROPICAL FORESTRY CHINESE ACAD OF FORESTRY
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