Marigold pollen differential medium and differentiation culture method

A technology of differentiation medium and culture method, which is applied in the field of marigold pollen differentiation medium and differentiation culture, can solve the problems of low callus induction rate, inability to accurately judge, low haploid, etc., and improve the differentiation rate of green shoots , the effect of promoting callus differentiation

Active Publication Date: 2019-12-10
济南易通城市建设集团股份有限公司
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AI Technical Summary

Problems solved by technology

Although the research on marigold anther culture has been reported, there are problems such as low callus induction rate, low differentiation rate, and low plant regeneration rate.
And in the regenerated plants obtained from anther culture, diploids account for the majority, aneuploids still have a small part, an...

Method used

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  • Marigold pollen differential medium and differentiation culture method
  • Marigold pollen differential medium and differentiation culture method
  • Marigold pollen differential medium and differentiation culture method

Examples

Experimental program
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Effect test

Embodiment 1

[0028] A kind of marigold pollen callus differentiation culture method, comprises the steps:

[0029] 1) Two marigold varieties 'Antigua' and 'Discovery' were used as test materials, and the pollen microspores were isolated for induction culture. When the callus grew to a size of 2-3 mm, the callus was inoculated into The light and dark cultures were alternately cultivated on the differentiation medium. The temperature of the light culture was 25°C, the light time was 12h / d, the light intensity was 1500lx, and the temperature of the dark culture was 22°C. After 3~4 weeks of culture, the callus differentiated into test-tube plantlets , when culturing for 30 days, the green shoot differentiation rate was counted, and the green shoot differentiation rate (%)=the number of calli that differentiated into green shoots / the total number of inoculated calli × 100%;

[0030] The formulation of the differentiation medium used is: KNO 3 1800mg / L, NH 4 NO 3 1450mg / L, KH 2 PO 4 ·H 2 O...

Embodiment 5

[0050] Embodiment 5: Ploidy identification test of regenerated plants

[0051] Adopt flow cytometer to carry out ploidy identification to the regenerated plant that embodiment 1 obtains, and specific method is: take more than 1cm 2 The leaves of the plant were chopped with a sharp blade in a 55mm petri dish, and 400uL of extract (15mol / LTris-HCl (pH7.5), 80mol / LKCl, 20mol / LNaCl, 20mol / LEDTA-Na 2 , 15mol / L mercaptoethanol, 0.05% Triton X-100). Leave it for 1 minute, then filter it with a 50 μm microporous nylon mesh, collect the filtrate in a standard test tube for on-machine testing, then centrifuge at 1000r / min for 8 minutes, discard the supernatant, and then add 1mL of propidium iodide staining solution , stained in the dark and low temperature for 20 minutes, and then tested on the machine, the results are shown in Table 5.

[0052]

[0053] It can be seen from Table 5 that in the regenerated plants of 'Antigua', the haploid rate was 35.6%, and the diploid rate was 56....

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Abstract

The invention discloses a marigold pollen differential medium and a differentiation culture method. The medium comprises KNO3, NH4NO3, KH2PO4.H2O, MgSO4.7H2O, CaCl2.2H2O, MnSO4.4H2O, ZnSO4.7H2O, H3BO3, KI, Na2MoO4.2H2O, CuSO4.5H2O, CoCl2.6H2O, ferrous glycine, inositol, thiamine hydrochloride, pyridoxine hydrochloride, niacin, threonine, asparagine, retinol, an air-plant herb extraction solution,6-BA, NAA, abscisic acid, vorinostat, 5-aminolevulinic acid, maltose, betaine and agar. By means of culture using the medium and the method, the plant differentiation rate of 70% or above can be obtained, and a certain technical support is provided for establishment of a marigold pollen high-efficiency culture system.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to a marigold pollen differentiation medium and a differentiation cultivation method. Background technique [0002] marigold( Tagetes erecta L.) is an annual herbaceous plant of the genus Marigold in the Compositae family. It is native to Mexico. Its growth temperature is 15~20°C. In a high temperature environment above 30°C, the plants grow leggy and have few flowers. Below 10°C, it can grow but the speed slows down. The growth cycle is lengthened. Marigold is more resistant to drought and likes a warm, humid and sunny environment. It is a multifunctional and multipurpose plant. First of all, marigold has high ornamental value, not only can be cultivated in the open field to beautify streetscapes and courtyards, but also can be potted to arrange mobile flower beds. Marigolds can be divided into three types: high, medium and short according to their plant height. The sho...

Claims

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Application Information

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IPC IPC(8): A01H4/00
CPCA01H4/001A01H4/008
Inventor 李传传
Owner 济南易通城市建设集团股份有限公司
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