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Method for inducing dissociate microspore callus of eggplant and regenerating plant strain

A callus induction and callus technology, applied in the field of plant cultivation, can solve problems such as tissue interference, achieve the effects of uniform morphology, diverse plant types, and improved probability

Inactive Publication Date: 2009-02-25
INST OF VEGETABLE & FLOWERS CHINESE ACAD OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, there are still many problems to be solved in anther culture. Due to the interference of various parts of anther tissues, the culture products may not come from pollen, resulting in the production of somatic hybrids of different ploidy

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0031]A method for inducing eggplant free microspore callus and plant regeneration, comprising the following steps: (1) selecting anthers: selecting flower buds of eggplants and simendou at the full flowering stage on healthy plants growing under natural conditions, and passing through microscopic examination Observe the developmental stages of the microspores, and select the microspores at the uninucleate middle stage and the marginal stage. The corresponding external characteristics of the flower buds are that the corolla is 1-2 mm lower than the calyx and the corolla is 1-2 mm higher than the calyx. The anthers are yellow-green before and after the sepals are about to crack.

[0032] (2) Pre-cultivation of anthers: comprising the following steps:

[0033] a) Disinfect flower buds: use 70-75% alcohol to disinfect the surface of flower buds for 2 minutes, then soak in 6.5% sodium hypochlorite for 10-15 minutes, and finally wash with sterile water for 3 times, each time for 5 m...

Embodiment 2

[0054] A method for inducing eggplant free microspore callus and plant regeneration, comprising the steps of:

[0055] (1) Select anthers: select the flower buds of eggplant and Simendou at the full flowering stage on the strong and healthy plant under natural conditions, observe the development period of microspores by microscopic examination, select the microspores of the uninucleate middle stage and the marginal stage, corresponding The external features of the flower buds are that the corolla is 1-2 mm lower than the calyx to 1-2 mm higher than the calyx, and the anthers are yellow-green before and after the sepals are about to split.

[0056] (2) Pre-cultivation of anthers: comprising the following steps:

[0057] d) Disinfect flower buds: use 70-75% alcohol to disinfect the surface of flower buds for 2 minutes, then soak in 6.5% sodium hypochlorite for 10-15 minutes, and finally wash with sterile water for 3 times, each time for 5 minutes, absorb with sterile paper dry ...

Embodiment 3

[0077] A method for inducing eggplant free microspore callus and plant regeneration, comprising the steps of:

[0078] (1) Select anthers: select the flower buds of eggplant and Simendou at the full flowering stage on the strong and healthy plant under natural conditions, observe the development period of microspores by microscopic examination, select the microspores of the uninucleate middle stage and the marginal stage, corresponding The external features of the flower buds are that the corolla is 1-2 mm lower than the calyx to 1-2 mm higher than the calyx, and the anthers are yellow-green before and after the sepals are about to split.

[0079] (2) Pre-cultivation of anthers: comprising the following steps:

[0080] g) Disinfect flower buds: use 70-75% alcohol to disinfect the surface of flower buds for 2 minutes, then soak in 6.5% sodium hypochlorite for 10-15 minutes, and finally wash with sterile water for 3 times, each time for 5 minutes, absorb with sterile paper dry ...

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Abstract

The invention is a method of inducement of isolated microspore callus of eggplant and plant regeneration. The major procedures are as follows. Appropriate anther is selected for pre-cultivation. Then dissociation and collection of microspore are carried out. And then static shallow pan dark culture is carried out in fluid nutrient medium. After 20 to 30 days, callus emerges successively. Callus is taken for cultivation under sunlight after emergence. When the callus grows to the size with the diameter of 2 to 6 millimeters, callus is transferred to solid regeneration culture medium for further cultivation. After 30 to 45 days, callus begins to differentiate to form buds. When bud grows to size of 1 to 3 centimeters, callus is transferred to rooting culture medium. After 10 to 15 days, strong root is developed. By adopting the method, the probability of obtaining haploid is high and frequency of microspore callus genesis is high. The invention is not only the technological breakthrough of eggplant haploid breeding, the inducing system of isolated microspore for cultivation of embryoid can also be a good experimental system for carrying out research on plant cell differentiation and development, molecular marker and gene map.

Description

technical field [0001] The invention relates to a plant cultivation method, in particular to a haploid breeding method of eggplant. Background technique [0002] At present, in the eggplant breeding work, the selection of inbred lines is very important work, but there are often problems such as long regeneration period. Haploid breeding is one of the important breeding methods to solve this problem, which can greatly improve the efficiency of breeding selection, reduce the size of breeding groups, and shorten the breeding period. The application and significance of haploid in genetics and breeding: 1. Accelerate genetic purification and shorten the breeding period; 2. It can identify genetic segregation types from the gamete level and improve selection efficiency; 3. It can effectively use microbial genetic technology in higher plants; 4. Utilize The ploidy of pollen callus or plant tissue can induce ploidy plants; 5. Improve the utilization of distant hybridization in bree...

Claims

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Application Information

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IPC IPC(8): A01H4/00C12N5/04
Inventor 连勇刘富中陈钰辉徐涵孙振英马欣范适宋燕
Owner INST OF VEGETABLE & FLOWERS CHINESE ACAD OF AGRI SCI
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