A method of genetic transformation of lily musk based on shallow liquid culture
A genetic transformation method and liquid culture technology, applied in the field of genetic transformation of musk lily, can solve problems such as low transformation efficiency of gene gun method, and achieve the effects of being beneficial to callus induction, expanding surface area and improving transformation efficiency.
Active Publication Date: 2020-01-10
NANJING FORESTRY UNIV
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Problems solved by technology
The transformation efficiency of the gene gun method is low, and it is easy to generate chimeras
The cost of Agrobacterium-mediated method is low, and the transformation efficiency is high. However, the transformation of lily using Agrobacterium-mediated method is still in the exploratory stage.
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[0026] The present invention is further described in detail through the following examples, but the content of the present invention is not limited thereto.
[0027] As shown in the figure, a kind of musk lily genetic transformation method based on shallow layer liquid culture comprises the following steps:
[0028] (1) Pre-cultivation: the pre-medium is liquid MSO medium + picloram 4.0mg / L, pH is 5.8, the pre-medium has been prepared to a fixed volume, and the pre-medium is packaged in a high-pressure steam sterilizer at 121°C for 20min. In a petri dish (diameter 9 cm), the liquid culture medium is divided into about 6mL per dish in a clean bench, and it is better to just submerge the thin slices of the scales; Store in an environment with a temperature of 25°C, a humidity of 50% to 60%, a light intensity of 2000 lx, light / darkness = 16h / 8h, and culture on MSO solid medium containing 100g sucrose for subculture every 2 months , the callus induction rate of the middle scales ...
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Abstract
The invention discloses a method for genetic transformation of lily musk lily based on shallow liquid culture, including pre-cultivation, strain activation, Agrobacterium infection and co-cultivation, degerming and screening culture, differentiation and acquisition of resistant seedlings, and stereofluorescence microscopy Steps such as observation and detection; the present invention takes the scales of the musk lily 'Leishan' test tube bulb with stronger callus generation ability and faster callus proliferation speed as experimental material, adopts plant cell thin-layer section method, and uses shallow layer liquid culture as experimental material. Based on the study of genetic transformation of lily musk by Agrobacterium tumefaciens mediated method, the green fluorescent protein gene (GFP) was transferred into lily musk, and the regenerated plants were observed with an upright stereo fluorescence microscope. Research provides the basis.
Description
technical field [0001] The invention relates to the field of biotechnology, in particular to a method for genetic transformation of lily musk lily based on shallow liquid culture. [0002] technical background [0003] Liliaceae Lilium plants are mostly world-renowned ornamental flowers, suitable for potted plants, fresh cut flowers and garden greening, and also have edible and medicinal values, and are increasingly becoming the first choice for improving the quality of life. The main cut lily varieties on the market are mostly musk lily, oriental lily and Asiatic lily. Lily has strong regeneration ability, and scales, stem tips, leaves, pedicels, receptacles, etc. can be used as explants to redifferentiate through callus or directly induce regeneration of small bulbs or adventitious buds. However, most of the studies on tissue culture of lily are based on rapid propagation, and the research on the regeneration system for the purpose of genetic transformation has also been r...
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IPC IPC(8): C12N15/84C12N1/20A01H5/00A01H6/56
CPCC12N15/8205
Inventor 席梦利赵乙琏孙惠霖姜珊施季森
Owner NANJING FORESTRY UNIV