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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 eggplant breeding methods, the present invention provides a protoplast electrofusion method for eggplant breeding

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

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

[0036] Step 1. Preparation of protoplasts by enzymatic hydrolysis:

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

[0038] Put the obtained pretreated tissue slices into the enzymolysis solution according to the mass volume ratio of the tender leaves of the eggplant seedlings and the enzymatic solution, respectively seal the sealing film and place it at a certain temperature and rotation speed, and carry out the enzymatic treatment in the dark. hydrolyzed to obtain the enzymatic hydrolysis products of two kinds of parents respectively;

[0039] Step...

Embodiment 2

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

[0050] Step 1. Preparation of protoplasts by enzymatic hydrolysis:

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

[0052] In this embodiment, the CPW13M solution is prepared with a mannitol solution with a mass volume ratio of 13% as a solvent and CPW salt as a solute; Formulated for solutes; CPW salts 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 246.0mg / L, KI 0.16mg / L, CuSO 4 ·5H 2 O 0.025mg / L, pH 5.8.

[0053] Use the aqueous solution of CPW salt as a solvent to prepare an enzymolysis solu...

Embodiment 3

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

[0066] Step 1. Preparation of protoplasts by enzymatic hydrolysis:

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

[0068] In this embodiment, the CPW13M solution is prepared with a mannitol solution with a mass volume ratio of 13% as a solvent and CPW salt as a solute; Formulated for solutes; CPW salts 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 246.0mg / L, KI 0.16mg / L, CuSO 4 ·5H 2 O 0.025mg / L, pH 5.8.

[0069] The enzymolysis solution is prepared with the aqueous solution of CPW salt as a ...

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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
CPCC12N15/02C12N13/00C12N5/14
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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