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Method for screening new triploid germplasm from seedlings of white flesh loquat

A white sand loquat and triploid technology, which can be used in individual particle analysis, material inspection products, instruments, etc., can solve the problems of cumbersome process, low screening efficiency, time-consuming and laborious, etc.

Inactive Publication Date: 2020-02-04
NANJING AGRICULTURAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The identification of loquat triploids used to be mainly through root tip chromosome compression and chromosome counting under a microscope. This method is not only cumbersome, but also time-consuming and laborious, and the screening efficiency is low.

Method used

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  • Method for screening new triploid germplasm from seedlings of white flesh loquat
  • Method for screening new triploid germplasm from seedlings of white flesh loquat
  • Method for screening new triploid germplasm from seedlings of white flesh loquat

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0028] The screening of embodiment 1 nucleus separation liquid

[0029] 1.1 Test materials and instruments:

[0030] In October 2018, the annual and biennial Suzhou Dongshan Baisha loquat seedlings in Jiangsu Baisha Loquat Resource Garden (Jiangsu Taihu Evergreen Fruit Tree Technology Promotion Center) were used as test materials.

[0031] The flow cytometer adopts CytoFLEX from BECKMAN COULTER Company, and selects the PE channel for sample loading and detection.

[0032] 1.2 Flow cytometry identification:

[0033] Method I:

[0034] Extract configuration:

[0035] MgSO4 buffer configuration: configure the buffer to 10mmol L -1 MgSO4 7H2O, 50mmol L -1 KCl, 5mmol L -1 HEPES, a mixture of 0.25% (v / v) Triton X-100, wherein HEPES is first configured to 1mol L - 1 HEPES solution: 23.8g of HEPES was dissolved in 90ml of water, adjusted to pH=8.0 with NaOH, and then added with water to make up to 100mL.

[0036] Extract A: MgSO 4 Add 1% (w / v) polyvinylpyrrolidone PVP to the...

Embodiment 2

[0057] According to the method of the present invention, 924 Baisha loquat seedlings were screened for triploids. According to the morphological parameter standards recorded in Example 1, 17 suspected triploid Baisha loquat strains were initially screened out.

[0058] Take the young leaves of the suspected triploid Baisha loquat, wash them with sterilized double distilled water, and absorb the water with absorbent paper. Simply shred the leaves and put them in a centrifuge tube, avoiding leaf veins and other parts as much as possible. 1 / 3 of the tube, too much is not conducive to sample grinding, and too little is not conducive to extracting a sufficient amount of complete cells. Add 6-8 clean zirconia beads, add extract solution A to the two-milliliter mark, and add no less than 1.5 mL of extract solution. If the sample cannot be ground immediately after the addition, the centrifuge tube should be placed on ice. Adjust the sample grinding machine to 50Hz, adjust the sample ...

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Abstract

The invention discloses a method for screening new triploid germplasm from seedlings of white flesh loquat, and the method includes the following steps of (1) performing preliminary screening of triploids from the population of seedlings based on the morphological characteristics of triploids of the white flesh loquat through morphological observation, wherein those that meet the following conditions are suspected triploids of white flesh loquat; and (2) further identifying the ploidy of the suspected triploids of white flesh loquat by flow cytometry. The invention is simple in solution preparation, easy to start operation steps, high in screening success rate and reliable in results, and has obvious advantages compared with other methods currently applied to the flow-ploidy screening. Compared with the traditional chromosome counting technology, the screening efficiency can be improved by 4-5 times by adopting the method. Through the method of the invention, large-scale and rapid identification of triploid seedlings of white flesh loquat can be realized.

Description

technical field [0001] The invention belongs to the field of biological detection and relates to a method for screening new triploid germplasm from Baisha loquat seedlings. Background technique [0002] Baisha loquat is deeply loved by consumers because of its delicate meat and high sweetness. Suzhou, Jiangsu is one of the main production areas of Baisha loquat. Most varieties of Baisha loquat have many and large cores, which affects their eating experience. Breeding triploid Baisha loquat can effectively solve this problem, and it is of great significance to promote the development of loquat industry. Seedling selection is an important way for the cultivation of new varieties of loquat. Among the main cultivars of loquat at present: varieties such as Baiyu (Qi Zihong, 2001), Jiefangzhong (Li Bilian, 1999), Dawuxing (Chen Guihu, etc., 2002) are all obtained through seeding. Selected and bred. Among the several ways of loquat triploid breeding, seed selection is an importa...

Claims

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Application Information

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IPC IPC(8): G01N33/00G01N21/64G01N15/14
CPCG01N15/14G01N21/6428G01N33/0098G01N2015/1006
Inventor 王三红荣志豪
Owner NANJING AGRICULTURAL UNIVERSITY
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