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Tree Cell Engineering Lab: LICA

Site Orléans: INRAE - Centre Val de Loire - 2163, Avenue de la pomme de pin - CS 4001 Ardon - 45075 Orléans Cedex 2

 https://www6.inrae.fr/lica_eng/

The LICA is a technical platform inaugurated in 2017 on the Orleans site of the INRAE Val de Loire Center. It is intended to house in a single location and for the duration of studies involving the production and characterization of genetically modified or edited trees. It is a confined building of the L2S2 type that complies with biological safety regulations. Thus, LICA's vocation is to meet the research, training and service needs of public or private research teams involving the production, multiplication and characterization of genetically modified forest resources for experimental purposes and aiming to increase our knowledge of the physiology of forest trees and their capacity to adapt to different environmental constraints. 

LICA brings together the expertise of researchers, engineers and technicians and the equipment of the Integrated Biology Unit for the Development of Tree and Forest Diversity (BioForA) and the Orleans Genetics and Forest Biomass Unit (GBFOR) of INRAE.

LICA has a Charter of use and the services offered are presented on its website (https://www6.inrae.fr/lica/Demande-de-Collaboration).

The activities developed at LICA mainly concern :

• The multiplication and breeding of forest trees from in vitro culture. This activity benefits from laboratories, in vitro culture chambers and automated greenhouses.
• Genetic transformation by co-culture of Agrobacteria to carry out functional genomic studies mainly in poplar, a model species for forest trees. The proposed techniques allow ectopic expression, expression reduction (by RNA interference) and KO (by genome editing) of the targeted genes.
• Basic molecular biology techniques to select the most promising events on trees grown under controlled conditions.
• Phenotyping techniques to evaluate the effects of the modification of the expression of target genes. LICA is particularly well equipped to carry out microphenotyping, especially in wood, the emblematic tissue of trees, thanks to its equipment and skills in histology (fluorescence stereomicroscopy, fluorescence microscopy, laser confocal microcopy, etc.) and molecular imaging (immunolocalizations, in situ hybridization, infrared microspectroscopy).

A support team of four permanent employees, assisted by non-permanent employees, ensures the supervision of the various projects and technological developments carried out in the LICA technical platform. LICA is backed by a BioForA scientific team, an expert in the functional genomics of wood formation. Governance is ensured by the LICA Scientific Council (6 members, including 3 external members).

LICA's services are open to all public and private research actors. Any questions or requests for collaboration or access to the equipment and services offered by LICA should be addressed to Gilles Pilate. The conditions of access are described in the Charter of Use and on the LICA website. For the design, implementation and management of user projects, LICA undertakes :

• Provide training and supervision by qualified personnel.
• Offer efficient and operational installations and equipment.
• Make available the techniques associated with these materials.
• Regularly inform users of the progress of their project.
• To provide the results of experiments as soon as they are available.
• Respect the confidentiality of the nature and results of the projects carried out.

After examination by the Scientific Committee of the requests submitted concerning their feasibility, we propose the type of project that best meets the requests (collaborative research project, use of equipment by the applicant after training, service provided by the platform's staff). We provide the necessary health and safety supervision for access to and use of LICA laboratories and equipment, in accordance with current French regulations on GMOs.

Each approved project will be quoted.

Rates differ depending on the type of user:

• Marginal cost (cost of using labs/equipment and labor excluding public salary) for academic users.
• Full cost (cost of use of laboratories/equipment and manpower excluding public salary + structural charges) for private users.

The LICA has the following equipment:

• An in vitro culture laboratory equipped for the preparation and autoclaving of growth media, for transplantation under laminar flow hood and for the rearing of in vitro plants in culture chambers.
• A molecular biology laboratory equipped for cloning, microbiology (PSM bacteriological hood) and nucleic acid extraction.
• An experimental greenhouse type S2 (150 m2) with four units for acclimatization and tree breeding. A large unit (75 m2) with a height of 5-6m allows a good development of the aerial part of the trees. A pit of 1 meter depth is also available allowing a good development of the roots.
• A histology laboratory equipped to carry out sections of plant tissues, their stainings and their observations by various techniques of optical microscopy. For this purpose, the laboratory has a LEICA RM2155 rotary microtome and a Thermo HM 650 vibratome. The laboratory is also equipped with :
  • a Leica fluorescence stereo-microscope which allows the in vivo detection of fluorescent proteins of GFP type
  • an Olympus inverted microscope for observations of cell cultures or tissue sections in transmitted light and epifluorescence
  • a Zeiss confocal microscope for the realization of optical sections and the acquisition of images in multiple fluorescences.
• An infrared spectroscopy laboratory with hygrometry and temperature control equipped with
  • an infrared spectrometer allowing the acquisition of near (NIR) and medium infrared spectra (MIR, FT-MIR, ATR-FT-MIR).
  • an imager coupled to the spectrometer allowing to realize high resolution infrared images in near and medium infrared