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Method for inducing anther calluses of sainfoin and special culture medium of method

A technology of callus induction and callus, which is applied in horticultural methods, botanical equipment and methods, applications, etc., can solve the problems of poor repeatability and low frequency of anther induction, and achieve the effect of increasing induction rate and cultivating efficiency

Inactive Publication Date: 2019-09-10
郭根霞
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, there are few domestic and foreign literature reports on the cultivation of anther cultures, and there are problems such as low frequency of anther induction and poor repeatability, so there is still a certain distance from the establishment of a mature culture system.

Method used

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  • Method for inducing anther calluses of sainfoin and special culture medium of method
  • Method for inducing anther calluses of sainfoin and special culture medium of method
  • Method for inducing anther calluses of sainfoin and special culture medium of method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0024] 1.1 Materials

[0025] The two species of red bean grass tested are: common red bean grass and Caucasian red bean grass.

[0026] 1.2 Method

[0027] 1) Collection of inflorescences: In the early flowering stage of the red bean grass, the inflorescences of the red bean grass are collected at 08:00-09:00 in the morning on a sunny day when there is no dew, and then stained with DAPI (4,6-diamidino-2-phenylindole) Inflorescences in the middle and late stages of uninucleate pollen development were screened by the method as test materials.

[0028] 2) Inflorescence pretreatment and disinfection: rinse the inflorescences with tap water, put them into a petri dish covered with moist filter paper, and place them in a low temperature treatment at 4°C for 48 hours. During this period, spray water once a day to keep the inflorescences and filter paper always moist; The inflorescences were centrifuged at a speed of 2500r / min for 8 minutes; the pretreated inflorescences were place...

Embodiment 2

[0031] Embodiment 2: The effect of centrifugation on the induction of red bean herb anther callus

[0032] In the step of inflorescence pretreatment and disinfection, set no centrifugation as the control (CK), set 1000, 1500, 2000, 2500, 3000r / min 5 test groups for centrifugation, and the centrifugation time is 8min. Other operation methods and examples 1, the callus induction rates of the two varieties of Ormosia chinensis were counted, and the results are shown in Table 1.

[0033]

[0034] It can be seen from the results in Table 1 that compared with the control (CK), centrifugation at 1000-3000r / min can promote the induction of anther callus, and the callus induction rate has increased; when the centrifugal speed is 2500r / min min, the callus induction rate was the highest, which were 44.65% for common red bean grass and 40.49% for Caucasian red bean grass, which were significantly different from the control.

Embodiment 3

[0035] Example 3: Effect of gamma-aminobutyric acid on the induction of red bean herb anther callus

[0036] When the callus is induced and cultured, the preparation of other components is the same as that of the induction medium in Example 1, and the concentration of γ-aminobutyric acid is respectively 0, 2.0, 4.0, 6.0, 8.0, 10.0mg / L of 6 kinds of induction medium , inoculated the anthers of Ormosia japonica in the above induction medium for cultivation, and counted the callus induction rates of the two varieties of Ormosia japonica, and the results are shown in Table 2.

[0037]

[0038] From the results in Table 2, it can be seen that compared with the control (γ-aminobutyric acid concentration is 0), the addition of γ-aminobutyric acid at the test concentration in the induction medium can improve the induction of the anther callus of the red bean grass Effect, in the range of 2.0-6.0mg / L, as the concentration of γ-aminobutyric acid increases, the callus induction rate i...

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Abstract

The invention provides a method for inducing anther calluses of sainfoin and a special culture medium of the method, and belongs to the technical field of pasture biology. The method includes, firstly, carrying out low-temperature and centrifugal pretreatment on the screened sainfoin inflorescence, which is beneficial to promoting the formation of anther calluses; and then taking off the anthers and inoculating on the callus induction culture medium for culture to obtain calluses. The callus induction culture medium is optimized on the basis of a basic culture medium, appropriately improves the concentrations of KNO3, KH2PO4, vitamins, sucrose and the like, adjusts the proportion of conventional plant hormones, and adds L-proline, gamma-aminobutyric acid, cyclic guanosine phosphate, brassinolide, N-methylmorpholine, fulvic acid, butyl hydroxy anisole, dithiothreitol and an astragalus sinicus extract, and thus the induction rate of the sainfoin anther calluses is effectively improved, so that the sainfoin anther culture efficiency is improved.

Description

technical field [0001] The invention relates to the field of herbage biotechnology, in particular to a method for inducing callus of red bean grass anthers and a special culture medium thereof. Background technique [0002] Red bean grass (Onobrychis viciaefolia Scop), also known as donkey bean and donkey hi bean, is native to Europe and is a perennial herb of the family Fabaceae. Red bean grass likes warm and cool, dry climate, and has great plasticity to adapt to the environment. It is resistant to drought, cold, early frost, late autumn precipitation, and other unfavorable factors. grow on soil. There are more than 120 kinds of plants of the genus Ormosia in the world, most of which are wild species. There are wild species distributed in the Tianshan Mountains and the northern foothills of the Altai Mountains in Xinjiang, my country. Most of the red bean grass cultivated in my country is imported from other countries and regions, and is planted in 23 provinces and regio...

Claims

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Application Information

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IPC IPC(8): A01H4/00
CPCA01H4/001A01H4/005
Inventor 郭根霞
Owner 郭根霞
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