Agronomy Team:Don C. Wilkerson

Isaac Wong

Stephanie Akins

Nicotiana benthamianaNicotiana: A genus of short-leafed plants in the nightshade family including Nicotiana tabacum

Nicotiana benthamiana: A close relative of tobacco belonging to the same genus of short-leafed plants and used as a model organism

Nicotiana tabacum: The common tobacco plant cultivated worldwide to produce tobacco leaf for cigarettes

Grow Pods:

Production Protocol:

Nicotiana bentamiama plants are grown using a Standard Production Protocol (SPP) developed by GCON/Caliber Biotheraputics. This SPP has evolved through a series of platform optimization studies/trials/evaluations, designed to reach targeted expression levels for a broad range of monoclonal antibodies.

Seeding/Plug Production

Transplant 21 Days

Grow 10 Days

Infiltration

Post Infiltration 7 Days

Harvest

Standard Production Protocol

Total Production Time = 38 Days

Seeding:The SPP begins by direct seeding in to Styrofoam plug trays, each containing 240 rockwool plugs. Prior to seeding, plugs are pre-soaked with a pH adjusted nutrient solution to ensure optimum germination and growth. Following seeding, trays are placed in a controlled environment Pod at 25⁰C and 75% humidity. Supplemental light is provided 24 hours/day by LED fixtures, delivering 60 microMoles/second/m2 at tray height. All trays are sub-irrigated 3X/day at 8 hour intervals, using a pH adjusted, hydroponic nutrient solution.

90% of all seed germinate within 3 – 5 days. Seedlings (plugs) will remain in this environment for approximately 21 days.

21 day old plugs are then hand transplanted in to trays containing pre-dibbled sheets of an expanded polypropylene growing substrate (SuretoGrow). Each tray is fitted with a perforated lid, designed to hold the substrate/plants in place during vacuum infiltration. Each Grow tray contains 60 plants.

Transplanting:

Transplanted trays are then placed in a controlled environment Pod at 25⁰C and 60% humidity. Supplemental light is provided 24 hours/day by LED fixtures, delivering 120 microMoles/second/m2 at tray height. All trays are irrigated 3X/day at 8 hour intervals, using a pH adjusted, hydroponic nutrient solution. Plants are grown under these conditions for 10 days prior to vacuum infiltration.

Grow:

Infiltration:

Following vacuum infiltration, plants are placed in a post-infiltration (PI) Pod at 25⁰C and 60% humidity. Supplemental light is provided 18 hours/day by LED fixtures, delivering 120 microMoles/second/m2 at tray height. All trays are irrigated 3X/day at 8 hour intervals, using a pH adjusted, hydroponic nutrient solution. Plants are grown under these conditions for 10 days prior to harvest.

Post Infiltration:

Harvest: