Rooms 102 and 110 are designated areas for growing Arabidopsis. A 16 hour photoperiod is maintained in both rooms. Lighting cycles are adjusted for seasonal changes in temperature and photoperiod.

Room 102 - General Information:

Used for non transgenic material only. Do not transfer plants, soil, waste or waste containers between 102 and 110. Waste from 102 does not require autoclaving. Storage of supplies, equipment, or personal items is not permitted in the room. Benches are to be used for growing space only, unattended supplies or personal items will be disposed of. Placing anything other than bacterial treated, dome enclosed trays under benches is not permitted. Anything else will be disposed of. The door is to be closed but unlocked at all times. Pesticide treatments begin promptly at the scheduled time. Anyone present will be asked to exit.

Procedures and Cultures:

Soil Preparation:

Users must schedule for planting needs. Twelve hours are required between soil preparation and use. Users request greenhouse staff to fill growing containers with treated soil. Soil is treated at 2 PM Monday thru Thursday. Friday soil is prepared before 12:00. Soil cannot be used within 12 hours of treatment. Soil requested by 2 PM will be available for use the following day. All soil for greenhouse maintained plants must be chemically treated. This includes seed sowing and transplanting. Transporting lab or growth chamber grown plants in untreated soil to the greenhouse is not recommended.

Soil is to be treated and prepared by the greenhouse staff only.

The greenhouse staff will prepare treated soil for plants started in growth chambers if they are intended to be transferred to the greenhouse. Autoclaving of soil does not provide any protection from subsequent infestation or infection by harmful organisms. Sterile autoclaved soil placed on a greenhouse bench can be invaded by disease or pests within minutes. Only the greenhouse staff are to treat and prepare soil for greenhouse use. Pre treatments and Chemical applications are essential and necessary to maintain a greenhouse environment within acceptable disease and pest limits.

Preparing growing Containers:

Use only clean, disinfected containers and inserts. User selects desired container and inspects tray for serviceability. Trays should not have excessive drainage holes.

Trays with holes greater than 3 cm should be sealed with autoclave tape. Discard if tray cannot be repaired. Check to assure tape or markings from prior use have been removed. Entire tray area cannot be completely filled with inserts. A small area must remain open to allow for irrigation. Position inserts in tray so drainage hole is visible in the open area.

Apply tape with user name and date to tray. Cover with clear dome. Request greenhouse staff fill container with treated soil. After treatment, colored labels will be placed in the growing medium to indicate treatments. A note will be placed on the clear dome indicating when the tray is safe for use and the length of time within which it can be used. Soil not used within 3 days of treatment will be discarded promptly by the greenhouse staff. Over aged soil, tray and inserts will be disposed of without notice.

Transplanting / Sowing:

Remove label from dome. Tray is ready for use. Do not water, soil is sufficiently wetted. Additional watering will dilute the pesticide treatment.

Plant Care:

Greenhouse staff perform irrigation and fertilizer treatments daily. Plants are checked several times and irrigated with consideration to moisture content, daily weather conditions, light and humidity levels, timing of additional pesticide treatments, etc. All irrigation is best left to the greenhouse staff. User intervention with irrigation complicates, and can interfere with, an assortment of fertilizer and pesticide treatments. Trays are evaluated individually and treated accordingly. If users find irrigation frequencies inadequate notify greenhouse staff and provide specific instructions. Requests, questions, and comments are welcome.

Drainage Concerns:

Trays are used to stabilize and transport growing containers. They are not a convenient receptacle for trash or debris. Use the waste container present in the room for the disposal of all pipette tips, rouged seedlings and trash. Drainage holes are clogged by the accumulation of soil and debris. Do not place plants or plant parts in the tray when removing or thinning young seedlings.

Air Cons:

Air cons provide an environment conducive to the development of disease and insect infestation. Air cons limit pesticide application methods and materials available. They greatly increase labor involved in pesticide treatments and cleaning of materials. Inspect plants closely before enclosing in air cons. If treatment is required, request treatment before using. If infestation is noticed after plants are enclosed, request treatment. The effectiveness of the treatment is related to timing from infestation. Plants requiring treatment after enclosure may have liquid pesticide residue in the air con base. Gloves should be worn when handling air cons that have been treated with pesticides.

Identifying Treatments:

The use of colored labels assists in identifying pesticide and nutritional treatments performed on a large number of independently tended growing containers. This system prevents unnecessary and potentially harmful duplication of treatments and indicates future treatments that may be required. These are solely for use by the greenhouse staff. Do not move, remove, add, mix, write on, re-label, dispose of, apply tape to or otherwise tamper with colored labels. Colored labels are recycled as projects are completed.

If moving individual inserts between trays, the insert can ONLY be moved to a tray containing identical colored labels. If the insert being moved contains colored labels, then identical colored labels MUST be inserted into the original tray.

Prompt Harvest:

Crop cycles should be completed in the shortest possible time. Potential for infestation and insect population increase with time. As crop time increases so do insect and disease levels. Contaminated plants compromise all plants in the room. Seed must be harvested when it is mature. Any trays that have no user identification, that are mature, orphaned or abandoned will be discarded by the greenhouse staff without notice.

Recycling supplies:

When plants are harvested all supplies are to be washed immediately. Remove any tape or markings identifying prior use. Used supplies are washed by user. Inserts and white labels should be disposed of. Colored labels, pots, trays, domes and air cons should be reused as long as they are serviceable. Trays etc must be stacked to permit rapid drying. Nested or consolidated supplies will not dry. Supplies are to be washed in the room they were used in and dried on bench space allocated to user. There is not adequate room for drying used supplies in quantity. Immediate handling minimizes space required for drying supplies and the time required to accomplish this. The greenhouse staff will store all properly washed used supplies. Abandoned, unwashed supplies, unserviceable, and used supplies that are not properly positioned for drying will be disposed of without notice.

Insert stability:

Inserts and small pots must remain stable and upright when the tray is irrigated. Small individual uncontained pots drift and fall over when the tray is flooded. Placing small containers in a larger insert will allow for watering without disturbing the plants.

Watering:

All Arabidopsis are sub-irrigated. Any plants requiring watering must be in a tray or container that will slowly drain. Plants that are not in trays will not be watered.

Bacterial treatments:

After plants have been sprayed with bacteria the tray is covered with a dome to maintain high humidity and delay drying. Check plants before treating. Infested plants that are in a closed tray fall between needed pesticide treatments if the treatment is made the same day as pesticides are applied. Check plants 1 or 2 days before treating and request any needed pesticide treatments prior to treating with bacteria.

Seed Harvest:

Mature seed is shed freely and widely dispersed. Other trays are exposed to contaminant seed. Prompt harvest minimizes seed contamination. Harvest techniques and awareness of seed dispersal when harvesting also limit the problem. Seed needs to be harvested in the room it was produced. Any neglected, mature plants shedding seed that are not labeled for user ownership may be discarded without notice.

Transferring Plants:

As a technique for prevention of introducing pests, plants should not be moved from the greenhouse to the labs. Plant transfer policies are established by individual labs.

Room 110 - General Information:

Licensed for housing transgenic plants. Used for transgenic plants only. All soil, plants, plant material, containers and supplies must be autoclaved when projects are complete.

Plant Culture:

The same as room 102.

User Responsibilities:

• Place all waste in unmarked autoclave bags. Users supply bags.

• Containment of plants transported to and from greenhouse.

• Locating transgenic plants in room 110 only.

• Requesting properly prepared soil for greenhouse use.

• Prompt harvesting of plants and recycling of supplies.

• Leave room in condition it was entered.

• Maintain supply cabinet in clean, organized condition.

• Keep door closed and locked at all times

• Notify greenhouse staff if introducing infected or infested plants.

• Containing waste within the room.

• Occupy allocated space only.

• Notify greenhouse staff of cultural problems or deficiencies.

• Floor is kept free of equipment and supplies, supplies left on the floor

• Are subject to disposal.

• Greenhouse Staff Responsibilities:

• Prepare soil for greenhouse use.

• Daily plant care, watering and fertilizing.

• Monitor and regulate environmental conditions and controls.

• Provide all chemical pest and disease control operations.

• Storage of properly cleaned recyclable supplies.

• Assist with stocking hallway storage cabinet.

• Request service for facility deficiencies.

• Assist in maintaining room in safe, clean condition.

• Consultation for cultural concerns.