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Lilium tenuifolium efficient genetic transformation system based on somatic embryogenesis

A genetic transformation system, the technology of lily lily, which is applied in the field of bulbous flower biology, can solve the problems of unreported genetic transformation of lily lily, and achieve the effects of avoiding the generation of a large number of chimeras, good preservation effect, and reduced screening work

Active Publication Date: 2018-07-20
SHENYANG AGRI UNIV
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Problems solved by technology

Most of the current researches on Lilium tenifolia focus on taxonomy, biology, asexual reproduction technology, etc. Chen Lijing et al. (2013) used scales as explants to initially discuss the tissue culture technology of Lilium tenifolia. Genetic transformation has not been reported

Method used

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  • Lilium tenuifolium efficient genetic transformation system based on somatic embryogenesis
  • Lilium tenuifolium efficient genetic transformation system based on somatic embryogenesis

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example 1

[0034] Example 1 pair carrying the target gene pTCK- S1 Transformation of the plant binary expression vector of -Ri to obtain transgenic plants

[0035] pTCK- S1 -Ri is a plant expression vector constructed on the basis of pTCK303 vector, containing hygromycin phosphotransferase gene and GUS reporter gene, and the target band is 269 bp long. In this example, Lilium tenifolia was purchased from a nursery stock seed sales office, and all biochemical preparations in this example were purchased from the market.

[0036] 1. Obtaining of Embryogenic Callus

[0037] Select aseptic seedlings of Lilium microphylla with a small bulb diameter of 1-1.2 cm, and cut the scales into 0.5 cm 2 Small pieces, concave side up, inoculated on somatic embryo induction medium [MS + 1.0 mg·L -1 Picloram (Picloram) + 0.2 mg·L -1 NAA (naphthalene acetic acid)]. The temperature of the culture room is 25±1°C, and an air-tight sealing film is used to ensure that the humidity in the tissue culture b...

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Abstract

The invention relates to a lilium tenuifolium efficient genetic transformation system based on somatic embryogenesis. The system comprises the following process steps of embryogenic callus induction through a somatic embryogenesis technology system, screening antibiotic hygromycin and kanamycin sulfate screening at the proper concentration, agrobacterium infection solution preparation, infection and co-culture, resistant callus screening and subculture; resistant somatic embryo germination and seedling forming, transgene plant GUS tissue dyeing and PCR detection. The lilium tenuifolium embryogenic callus is used as a receptor material; the somatic embryo is generated through induction after the genetic transformation for the first time, and is further and directly developed into a completeplant; the conversion rate is obviously improved in comparison with other kinds of lilium tenuifolium in the existing report, and can reach 29.17 percent. The system has the advantages that the recipient cell cardinal number is great; the somatic embryo induced after the conversion is a single cell source; the generation of a chimera is well avoided; the transformation generation exogenous geneshereditary stability is high; the aberration rate is low.

Description

technical field [0001] The invention belongs to the field of bulbous flower biotechnology, and specifically relates to the establishment of a stable genetic transformation system for lily, that is, to establish a stable and efficient genetic transformation system for lily lily based on the Agrobacterium-mediated method using induced embryogenic callus as a receptor. transformation system. Background technique [0002] Plant genetic transformation is a unique DNA cross-kingdom transfer technology. It is a plant genetic improvement technology that specifically introduces foreign genes into the plant genome, allowing them to be inherited and stably expressed in offspring plants. This technology is the core of plant genetic engineering. The establishment and development of transgenic technology enables people to introduce specific genes into different plants for expression according to their needs, providing a new way for plant genetic improvement. lily( Lilium ) is an importa...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/82A01H5/00A01H4/00A01H6/56
CPCA01H4/00C12N15/8205
Inventor 孙红梅严瑞王志平王春夏李宏宇
Owner SHENYANG AGRI UNIV
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