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Method for inducing maize microspore to form embryoid

A technology of microspores and embryoids, applied in the field of genetic breeding, can solve problems such as complicated operation steps, unfavorable popularization and application, and not very mature, so as to improve the selection effect, speed up the breeding process, and simple operation

Inactive Publication Date: 2012-03-28
WUHAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The cultivation technology of microspores of gramineous crops was carried out relatively late, and it was not very mature. There were reports on the cultivation of microspores of maize (Isolated microspore culture of maize: effects of isolation technique, reduced temperature, and sucrose level. Plant Cell Rep. ,1990,8:628-631), but it needs to use a special instrument (Waring MC-2), the operation steps are complicated, and it also increases the chance of pollution, which is not conducive to the popularization and application of this technology

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0028] 1) When corn grows in the field to the big bell mouth period (about 50 days), select the robust growth and late mononuclear tassels, wrap the tassels with plastic wrap, and cool them at 5°C for 7 days;

[0029] 2) Peel off the spikelets from the tassels, immerse them in 75% alcohol (volume ratio) for 30 seconds, and then use 0.1% Hgcl 2 The solution (mass to volume ratio) is sterilized for 10 minutes, washed with sterile water for 3 times, then transferred to a centrifuge tube, and the extract is pre-cooled at about 4°C. The composition of the extract is 150 mg / l (NH 4 ) 2 SO 4 +3000 mg / l KNO 3 +150 mg / l CaCl 2 ·2H 2 O + 450 mg / l MgSO 4 ·7H 2 O +600 mg / l KH 2 PO 4 +60 g / l sucrose+2.5 g / l 3-(N-morpholine) propanesulfonic acid, add KOH to adjust the pH to 6.0. The spikelet was squeezed with a glass rod to obtain a microspore suspension, filtered through a 100-mesh stainless steel sieve, and centrifuged at 100 g for 3 min. After the supernatant was discarded, the extract ...

Embodiment 2

[0037] 1) When corn grows to the big bell mouth period (about 50 days) in the field, select the robust growth and late mononuclear tassels, wrap the tassels with plastic wrap, and cool them at 8°C for 14 days;

[0038] 2) Peel off the spikelets from the tassels for surface disinfection (first immerse in 75% alcohol (volume ratio) for 30s, then use 0.1% Hgcl) 2 Sterilize the solution (mass to volume ratio) for 10 minutes), rinse with sterile water for 3 times, transfer to a centrifuge tube, and add the pre-cooled extract at about 4°C. The extract is 150 mg / l (NH 4 ) 2 SO 4 +3000 mg / l KNO 3 +150 mg / l CaCl 2 ·2H 2 O + 450 mg / l MgSO 4 ·7H 2 O +600 mg / l KH 2 PO 4 +60 g / l sucrose+2.5 g / l 3-(N-morpholine) propanesulfonic acid, add KOH to adjust the pH to 6.0. The spikelet was squeezed with a glass rod to obtain a microspore suspension, filtered through a 100-mesh stainless steel sieve, and centrifuged at 100 g for 3 min. After the supernatant was discarded, the extract was added for...

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Abstract

The invention relates to a method for inducing maize microspore to form embryoid, and belongs to the field of plant breeding. The method comprises the main steps of: selecting proper tassel for sterilization; extruding spikelet in extracting solution, sieving, and centrifuging to collect the microspore; inoculating into an induction medium, and performing heat shock treatment in an oven with the temperature of between 32 and 36 DEG C for 24 to 48 hours; and putting into a culturing room with the temperature between 25 and 28 DEG C, and performing culture under dark on a shaker for 20 to 25 days to form the embryoid. The method for inducing the maize microspore to form the embryoid has the advantages of high embryoid formation rate, short culture period, simple operation and the like. Whenthe method is used for maize inheritance breeding, the breeding course can be quickened greatly, and the selection effect is obviously improved.

Description

Technical field [0001] The invention relates to a method for inducing corn microspores to form embryoid bodies, and belongs to the field of genetic breeding. Background technique [0002] Maize is one of the main food crops in my country. Because it is a cross-pollinated crop, it takes 5-6 years to select an inbred line through conventional breeding, which consumes a lot of manpower and material resources, and sometimes its homozygosity cannot be guaranteed. Haplotypes can not only shorten the breeding period, but also improve the purity, which has great application value. Obtaining homozygous double haploid plants through anther culture has been widely used in crop variety improvement. The microspore culture technology developed on the basis of anther culture is arousing great interest of many researchers. [0003] Microspores are a brief but important stage in the development of male gametophytes in the life history of higher plants, including cells from the dissociation of the ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A01H4/00C12N5/04
Inventor 李立家何世斌李慧谭珺隽张琦
Owner WUHAN UNIV
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