A kind of method of obtaining the pseudo-thrift grass mutant
A technology for the generation of mutants of Pseudomonas pratense, which is applied in the field of obtaining mutants of Pseudomonas spp., can solve the problems of low-temperature plasma treatment of plant somatic cell mutation breeding, etc., and achieve the goals of shortening the breeding cycle, low cost, and accelerating the breeding process. Effect
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[0034] The method for obtaining the mutant of C. chinensis of the present invention comprises taking the callus of C. sylvestris and performing low-temperature plasma mutagenesis to obtain the mutagenized callus. The present invention has no special limitation on the type and source of the sage grass, and the conventional sage grass in the field can be used. The method for preparing the callus of P. chinensis according to the present invention preferably comprises: taking the sterilized young ears of P. spp. ; The induction medium uses MS medium as the basic medium, and also includes 1-5 mg / L of 2,4-dichlorophenoxyacetic acid, 0.04-0.2 mg / L of 6-benzylaminopurine, and 20-40 g / L of sucrose And plant gel (fixed support callus) 1 ~ 5g / L. The young ears of C. chinensis in the present invention are preferably the young ears of C. chinensis after the leaf sheaths have been removed. The method of the removal is not particularly limited in the present invention, and conventional meth...
Embodiment 1
[0049] Take the fresh young ears of Pseudomonas chinensis strain 'Common', peel off the leaf sheaths, disinfect and sterilize with 75% alcohol, and then inoculate them into the induction medium (the medium formula is: MS medium, add 2,4-dichlorophenoxy Acetic acid 3mg / L and 6-benzylaminopurine 0.1mg / L, additional sucrose 30g / L, phytogel 3g / L, pH 5.8), cultivated in dark at 25±1°C for 3 weeks, to obtain light yellow, granular embryogenic callus.
[0050] Put 20 embryogenic calli with a diameter of about 0.5 cm in a glass petri dish, seal it with a parafilm, place it in a low-temperature plasma processor, and accept low-temperature plasma treatment generated by glow discharge. The processing parameters are as follows: the processing power is 300W, the processing time is 90s, and the medium is a mixed gas of helium and air (He:air=8:1).
[0051] The embryogenic callus after the low-temperature plasma treatment was inoculated into the differentiation medium (the formula of the me...
Embodiment 2
[0056] Take the fresh young ears of Tifblair variety 'Tifblair', peel off the leaf sheaths, disinfect and sterilize with 75% alcohol, and then inoculate them into the induction medium (the medium formula is: MS medium, add 2,4-dichlorophenoxy Acetic acid 3mg / L and 6-benzylaminopurine 0.1mg / L, additional sucrose 30g / L, phytogel 3g / L, pH 5.8), cultivated in dark at 25±1℃ for 3 weeks to obtain light yellow, granular embryogenic callus.
[0057] Put 20 embryogenic calli with a diameter of about 0.5 cm in a glass petri dish, seal it with a parafilm, place it in a low-temperature plasma processor, and accept low-temperature plasma treatment generated by glow discharge. The processing parameters are as follows: the processing power is 400W, the processing time is 90s, and the medium is a mixed gas of helium and air (He:air=8:1).
[0058]The embryogenic callus after the low-temperature plasma treatment was inoculated into the differentiation medium (the formula of the medium was: MS ...
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