|Table of Contents|

Establishment of Efficient Transgenic System and Introduction of EPSP Gene Into Lilium

《北方园艺》[ISSN:1001-0009/CN:23-1247/S]

Issue:
2023年04
Page:
46-53
Research Field:
Publishing date:

Info

Title:
Establishment of Efficient Transgenic System and Introduction of EPSP Gene Into Lilium
Author(s):
ZHANG Jie; GUO Wenjie; CAI Xuanmei
(Biotechnology Institute，Fujian Academy of Agricultural Sciences，Fuzhou,Fujian 350003)
Keywords:
Lilium; EPSP gene; transformation; transgene
PACS:
-
DOI:
10.11937/bfyy.20222378
Abstract:
Ultrathin sections (0.5-1.0 mm) of aseptic bulbs of Lilium orientalis ‘Siberia’ were used as test materials.Callus induction culture conditions were screened by PIC and 2，4-D at different concentrations.The tolerance to Cef，Kan and Hgy was tested by direct section differentiation and callus differentiation.The effects of different drying pretreatment，MES and AS on the transformation effect were analyzed，and the positive rate of gene transformation and transformation effect were verified by the introduction of EPSP gene,in order to optimize the technical parameters of Lilium genetic transformation，establish an efficient genetic transformation system and introduce EPSP.The results showed that PIC had a good effect on callus induction.MS+1 mg?L-1 PIC+0.3 mg?L-1 NAA was the best medium for callus induction.Antibiotic sensitivity test showed that 75 mg?L-1 Kan or 20 mg?L-1 Hyg combined with 250 mg?L-1 Cef was suitable for resistance screening.The positive rates of ultrathin scales and callus were significantly increased after drying and preculture for 60 minutes and 30 minutes.MS+10 mmol?L-1 MES+200 μmol?L-1 AS treated with heavy suspension，the ultra-thin scales obtained an efficient conversion rate of 48.3%，which was better than callus transformation.At the same time，EPSP gene was transformed into Lilium，and molecular detection indicated that transgenic positive strains had been obtained.In this study，two sets of efficient genetic transformation systems of ultra-thin section and callus as the receptor were established，and EPSP gene was successfully introduced to obtain glyphosate resistance of 800 mg?L-1，laying a technical foundation for the subsequent production and application of transgenic technology.

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Last Update: 2023-03-21