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Cultivation method for barley microspores

A culture method, technology of microspores, applied in the field of agricultural biology, to achieve the differentiation potential of large green seedlings, and to promote the formation and differentiation of green seedlings

Inactive Publication Date: 2019-01-01
SHANGHAI ACAD OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0008] The purpose of the present invention is to provide a method for cultivating barley microspores. By performing low-temperature treatment on isolated florets, the callus yield and the amount of green shoots in the culture of barley microspores are improved, the regeneration ability is greatly improved, and the number of donor materials is solved. Less problems, simple operation, cost saving

Method used

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  • Cultivation method for barley microspores
  • Cultivation method for barley microspores
  • Cultivation method for barley microspores

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Experimental program
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Embodiment

[0030] 1. Materials

[0031] Three materials of barley 6-3-10, 6-3-20 and 6-3-23 were planted in pots in an artificial climate chamber with a light of 25 μmol m -2 ·s -1 , 12h·d -1 , humidity 70%, temperature set to light 22 dark 18 ℃.

[0032] 2. Method

[0033] 2.1 Material collection and ear pretreatment

[0034] Select the ears whose small flowers and microspores in the middle are in the early-middle stage of uninucleation, keep the base of the young ears at a length of 1.5cm, cut off the leaves, wrap them in wet gauze, put them in a fresh-keeping bag, and put them in a refrigerator at 5°C for low-temperature pretreatment for 2-3 weeks .

[0035] 2.2 Pretreatment of florets

[0036] Before inoculation, on the ultra-clean workbench, sterilize with 10% sanitin for 10 minutes, rinse with sterile water 4 to 5 times, pick 5 small flowers of tassels in each test tube (50ml), pour 12ml of extract, pH 5.8, and place 0d, 1d and 2d were treated in the refrigerator at 5°C, and...

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Abstract

The invention discloses a cultivation method for barley microspores. The cultivation method comprises the following steps of selecting young spikes, performing pretreatment, taking florets on the young spikes, performing disinfection, washing the florets with sterile water, performing inoculation to an extracting solution, and performing treatment for 1-2 d at 1-10 DEG C; performing rotary cuttingon the florets after inoculation, performing filtering and centrifugation, and collecting microspores; putting the collected microspores in the extracting solution for 1-3 d in the dark at 24-26 DEGC, and purifying the microspores with maltose; and washing the purified microspores with an induction medium, performing suspension, regulating the density of the microspores, performing inoculation to a culture dish, performing sealing, and performing cultivation for 15-24 d in the dark at 24-26 DEG C. In the cultivation method, by performing low temperature treatment on the florets in vitro, thecallus yield and the seedling formation amount in cultivation of the barley microspores are increased, the regeneration capacity is greatly improved, the problem of less amount of a donor material issolved, the operation is simple and feasible, and the cost is saved.

Description

technical field [0001] The invention belongs to the field of agricultural biotechnology, and in particular relates to a method for cultivating barley microspores. Background technique [0002] Plant microspores (Microspore) are haploid cells, which are produced by the tetrads formed by pollen mother cells after meiosis. Under in vitro stress conditions, they can switch from the original gametophyte development pathway to the sporophyte development pathway. , through embryonic development, forming a complete plant. [0003] Since haploid plants become double haploid plants (DHs) after chromosome doubling, alleles will not be lost due to segregation in subsequent generations, which can avoid serious segregation of hybrid offspring, thereby accelerating the process of trait recombination and shortening The number of years of breeding enables various traits to be displayed, so that good genes can be screened at the gametophyte level, which greatly improves the selection efficie...

Claims

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Application Information

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IPC IPC(8): A01H4/00
CPCA01H4/001A01H4/008
Inventor 郭桂梅陈志伟何婷陆瑞菊刘成洪黄剑华
Owner SHANGHAI ACAD OF AGRI SCI
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