Protoplast electrofusion method for eggplant breeding

A protoplast and electrofusion technology, applied in biochemical equipment and methods, fusion cells, and botanical equipment and methods, etc., can solve the problems of long cycle, poor predictability, low efficiency, etc. The effect of enhancing regenerative activity

Pending Publication Date: 2020-12-11
HARBIN CITY ACAD OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] In order to solve the problems of long period, low efficiency and poor predictability of the existing eg

Method used

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Examples

Experimental program
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Example Embodiment

[0034] Example 1

[0035] A protoplast electrofusion method for eggplant breeding, comprising the steps of:

[0036] Step 1. Enzymatic hydrolysis to prepare protoplasts:

[0037] Take the tender leaves of 1# eggplant seedlings and the tender leaves of 2# eggplant seedlings as parents to prepare protoplasts, cut the tender leaves of the seedlings into tissue pieces of the same size and place them in CPW13M solution for pretreatment for 30min respectively, filter to remove the CPW solution, and the tissue The slices were washed in CPW9M solution for 2 to 3 times;

[0038] Put the pretreated tissue pieces into the enzymolysis solution according to the mass-volume ratio of the young leaves of the eggplant seedlings to the enzymolysis solution, respectively seal the parafilms and place them at a certain temperature and rotation speed. solution to obtain two parental enzymatic hydrolysis products;

[0039] Step 2, separation and purification of protoplasts:

[0040] The enzymoly...

Example Embodiment

[0048] Example 2

[0049] A protoplast electrofusion method for eggplant breeding, comprising the steps of:

[0050] Step 1. Enzymatic hydrolysis to prepare protoplasts:

[0051] Take the tender leaves of 1# eggplant seedlings and the tender leaves of 2# eggplant seedlings as parents to prepare protoplasts, cut the tender leaves of the seedlings into tissue pieces of the same size and place them in CPW13M solution for pretreatment for 30min respectively, filter to remove the CPW solution, and the tissue The slices were washed in CPW9M solution for 2 to 3 times;

[0052] In this embodiment, the CPW13M solution is prepared by using a mannitol solution with a mass-volume ratio of 13% as a solvent and CPW salt as a solute; the CPW9M solution is prepared with a mannitol solution with a mass-volume ratio of 9% as a solvent, with CPW salt Formulated for solutes; CPW salts are formulated as KH 2 PO 4 27.2mg / L, KNO 3 101.0mg / L, CaCl 2 ·2H 2 O 1480.0mg / L, MgSO 4 ·7H 2 O 246.0...

Example Embodiment

[0064] Example 3

[0065] A protoplast electrofusion method for eggplant breeding, comprising the steps of:

[0066] Step 1. Enzymatic hydrolysis to prepare protoplasts:

[0067] Take the tender leaves of 1# eggplant seedlings and the tender leaves of 2# eggplant seedlings as parents to prepare protoplasts, cut the tender leaves of the seedlings into tissue pieces of the same size and place them in CPW13M solution for pretreatment for 30min respectively, filter to remove the CPW solution, and the tissue The slices were washed in CPW9M solution for 2 to 3 times;

[0068] In this embodiment, the CPW13M solution is prepared by using a mannitol solution with a mass-volume ratio of 13% as a solvent and CPW salt as a solute; the CPW9M solution is prepared with a mannitol solution with a mass-volume ratio of 9% as a solvent, with CPW salt Formulated for solutes; CPW salts are formulated as KH 2 PO 4 27.2mg / L, KNO 3 101.0mg / L, CaCl 2 ·2H 2 O 1480.0mg / L, MgSO 4 ·7H 2 O 246.0...

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Abstract

The invention relates to a protoplast electrofusion method for eggplant breeding, and belongs to the technical field of plant breeding. In order to solve the problems of long period, low efficiency and poor predictability of an existing eggplant breeding method, the invention provides the protoplast electrofusion method for eggplant breeding. The method comprises the following steps: respectivelycarrying out enzymolysis on eggplant donor and receptor tissues to prepare protoplasts, separating and purifying the protoplasts, and respectively carrying out passivation treatment, fusion promotiontreatment and electrofusion treatment on the donor and receptor protoplasts. Hybrid cells formed by electrofusion of the passivated donor and receptor can recover division to form calluses because thephysiological metabolism is complementary, and only the hybrid cells can obtain regenerated plants. The screening system is simple and time-saving, the obtained hybrid plants are parent genes, no unsafe factors exist, and thereby popularization and use are easy to carry out. According to the method, a fusion rate of two parents of eggplants can be remarkably improved, and the efficient transfer of excellent quality characteristics such as disease resistance and stress resistance of eggplants is realized.

Description

technical field [0001] The invention belongs to the technical field of plant breeding, in particular to a protoplast electrofusion method for eggplant breeding. Background technique [0002] Eggplant is a fruit and vegetable, rich in nutrition, high in protein, vitamins and various trace elements. It is the main vegetable variety in summer and autumn in northern my country. However, some original local varieties are seriously degraded, low in yield, and poor in quality. It is difficult to adapt to the needs of greenhouses, film-covered and other protected areas and the quality requirements of consumers. Therefore, it is necessary to seek early maturity, high quality, high yield, and resistance New eggplant varieties with strong sex are the top priority of current breeding work. [0003] The conventional breeding technology currently used in breeding has the following three defects: first, the breeding cycle is long, and it generally takes seven or eight years or even more t...

Claims

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Application Information

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IPC IPC(8): C12N15/05C12N13/00C12N5/14
CPCC12N5/14C12N13/00C12N15/02
Inventor 李烨姜景彬曹必好李炜许向阳李景富李宇
Owner HARBIN CITY ACAD OF AGRI SCI
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