Best PracticesBest Practices
For researchers using For researchers using Biotech Support Facility & Rightmire Hall Biotech Support Facility & Rightmire Hall Growth Chambers and GreenhouseGrowth Chambers and Greenhouse
IntroductionsIntroductions Gary Posey, Greenhouse SuperintendentGary Posey, Greenhouse Superintendent
Day to Day at BSF & RightmireDay to Day at BSF & Rightmire posey.8 292-9241posey.8 292-9241
Joan Leonard, Program ManagerJoan Leonard, Program Manager Day to Day at Bio Sci GreenhouseDay to Day at Bio Sci Greenhouse Oversee renovations and implementation of Oversee renovations and implementation of
best practices at BSF & Rightmirebest practices at BSF & Rightmire leonard.4 292-7904leonard.4 292-7904
Why?Why?
1.1. Reduce/ eliminate pest and disease Reduce/ eliminate pest and disease issues & improve plant healthissues & improve plant health
2.2. Increase experiment successIncrease experiment success3.3. Improve uniformity and repeatabilityImprove uniformity and repeatability4.4. Abide by all regulations set by NIH, Abide by all regulations set by NIH,
APHIS, EPA, ODA and Federal lawsAPHIS, EPA, ODA and Federal laws
Planting & Growing Planting & Growing ““DoDo’’ss ””
DO use DO use clean and sterile clean and sterile pots and trayspots and trays DO DO labellabel all pots/flats with researcher name -unlabeled plants will all pots/flats with researcher name -unlabeled plants will
be considered abandoned and will be disposed of.be considered abandoned and will be disposed of. DO surface sterilize seed (10% bleach, 10 min.)DO surface sterilize seed (10% bleach, 10 min.) DO follow DO follow order of entry order of entry to chambers/growth rooms/greenhouseto chambers/growth rooms/greenhouse DO water and fertilize properly DO water and fertilize properly
(use care (use care not to overwaternot to overwater)) DO wear dedicated DO wear dedicated lab coat lab coat for each growing spacefor each growing space DO know how to ID thrips, fungus gnats and signs of INSVDO know how to ID thrips, fungus gnats and signs of INSV Do make sure all staff and students have read the best Do make sure all staff and students have read the best
practices and met with Gary for a training session.practices and met with Gary for a training session.
Potting Room 100 BSFPotting Room 100 BSF No seeds are permitted in the potting No seeds are permitted in the potting
area, room 100 BSF to prevent area, room 100 BSF to prevent contamination of mediacontamination of media
Fill pots in 100 BSF then plant seeds in Fill pots in 100 BSF then plant seeds in room 160A, the greenhouse or your labroom 160A, the greenhouse or your lab
For Arabidopsis, 3 scoops of media = 1 For Arabidopsis, 3 scoops of media = 1 flat. Incorporate 2 TBL of osmocote for flat. Incorporate 2 TBL of osmocote for each flat prepared.each flat prepared.
Photo courtesy of Purdue University
Proper WateringProper Watering Arabidopsis IS NOT AQUATIC!Arabidopsis IS NOT AQUATIC! Overwatering: #1 cause of plant diseases, pest problems Overwatering: #1 cause of plant diseases, pest problems
and plant death! Too much-Too often.and plant death! Too much-Too often. Overwatering weakens roots and allows pests and Overwatering weakens roots and allows pests and
diseases an opportunitydiseases an opportunity NeverNever leave plants in a tray of standing water. Soil will leave plants in a tray of standing water. Soil will
reach saturation within 10 minutes—excess water in reach saturation within 10 minutes—excess water in trays after this will only weaken the plants.trays after this will only weaken the plants.
Green algae growing on soil surface is sure indication of Green algae growing on soil surface is sure indication of overwatering.overwatering.
Plants found standing in trays of water will have holes Plants found standing in trays of water will have holes poked into the trays!poked into the trays!
QuantityQuantity Watch nested flats: water can get Watch nested flats: water can get ““trappedtrapped”” Only water pots/cells that are dry instead of whole flat.Only water pots/cells that are dry instead of whole flat. Know if plants need water—what is dry?Know if plants need water—what is dry? If bottom watering whole flat, add only enough water to If bottom watering whole flat, add only enough water to
cover bottom of flat. This should get taken up within a cover bottom of flat. This should get taken up within a few minutes. If not, then plants were not thirsty.few minutes. If not, then plants were not thirsty.
Pour off excess water after 10 minutes! Pour off excess water after 10 minutes! Never leave standing in water longer than 30 minutes. Never leave standing in water longer than 30 minutes.
Water not used in 30 minutes will deprive roots of Water not used in 30 minutes will deprive roots of oxygen.oxygen.
FrequencyFrequency Check daily BUT Check daily BUT only water as needed!only water as needed! Water individuals when dry rather than watering all Water individuals when dry rather than watering all
plants in flat at same timeplants in flat at same time DonDon’’t follow a rigid watering schedulet follow a rigid watering schedule Avoid Avoid ““watering insurancewatering insurance””
This is overwatering to compensate for not checking on plants This is overwatering to compensate for not checking on plants daily.daily.
Heavy watering on Friday will make sure plants donHeavy watering on Friday will make sure plants don’’t need any t need any over weekend and possibly even until following Friday or over weekend and possibly even until following Friday or longer!longer!
No need to overwater to make it through the weekend. A well-No need to overwater to make it through the weekend. A well-watered plant on Friday will be fine on Monday.watered plant on Friday will be fine on Monday.
Rarely have we seen underwatered plants! This is easily fixed Rarely have we seen underwatered plants! This is easily fixed by adding water. Overwatering cannot be easily fixed.by adding water. Overwatering cannot be easily fixed.
Disposal Disposal ““DODO’’ss””
DO cover all plants during transport and disposalDO cover all plants during transport and disposal DO DO promptlypromptly dispose of unnecessary plants in dispose of unnecessary plants in yellowyellow
steam carts (no plastics, no stakes, etc)steam carts (no plastics, no stakes, etc) DO keep growing spaces and adjacent areas, and DO keep growing spaces and adjacent areas, and
potting room 100 clear of debris/dirty pots/dead plantspotting room 100 clear of debris/dirty pots/dead plants DO report infestations of pests and diseases to DO report infestations of pests and diseases to
managermanager DO immediately discard all INSV infected plantsDO immediately discard all INSV infected plants DO make sure all lab staff are educated in DO make sure all lab staff are educated in
procedures and are following them. This is the procedures and are following them. This is the responsibility of the PI.responsibility of the PI.
Handling Handling ““DONDON’’TsTs””
DONDON’’T leave chamber/rooms doors T leave chamber/rooms doors hanging openhanging open
DONDON’’T move plants from one T move plants from one chamber/room to anotherchamber/room to another
““DONDON’’T put all your eggs in one basket.T put all your eggs in one basket.”” (Stagger germination dates and location (Stagger germination dates and location of valuable plants to increase chances of of valuable plants to increase chances of success.)success.)
Thrips IDThrips ID Western Flower ThripsWestern Flower Thrips
Adults are less than 1/20th of an inch (<2mm) long
Eggs are laid in epidermis on underside of leaf
Only larval stages acquire INSV while feeding
Pupal stages occur in soil
Adults spread INSV when feeding
Fungus Gnat IDFungus Gnat ID
Larvae feed on plant roots & spread diseases
Slows plant growth & can kill seedlings
Thrive in wet soils
Best control is prevention through proper watering—do not overwater
Powdery MildewPowdery Mildew
Common in crowded conditions with poor air circulation, low light and dampness—AKA a growth chamber!
Spores are short-lived and need moisture to germinate.
Sleeved plants are especially susceptible—cut holes in sleeves for better air flow.
Water carefully!
INSV in ArabidopsisINSV in Arabidopsis Dead LeavesDead Leaves
INSV in ArabidopsisINSV in Arabidopsis Yellow Mid-RibYellow Mid-Rib
INSV in ArabidopsisINSV in Arabidopsis Purple LeavesPurple Leaves
INSV in ArabidopsisINSV in Arabidopsis Random DeathRandom Death
INSV HandlingINSV Handling Plants testing positive for INSV will be immediately Plants testing positive for INSV will be immediately
removed from growth spaces and removed from growth spaces and destroyed.destroyed. No new plants will be permitted in an infected No new plants will be permitted in an infected
space. space. Any plant to be removed from an infected space Any plant to be removed from an infected space
must be bagged before removal. must be bagged before removal. Anyone working in an INSV infected space is to Anyone working in an INSV infected space is to
immediately exit the building when work is immediately exit the building when work is completed- they may return the next day.completed- they may return the next day.
BeneficialsBeneficials N. cucumerisN. cucumerisFeeds on Thrips larvaeFeeds on Thrips larvae
BeneficialsBeneficials Hypoaspis milesHypoaspis milesFeeds on Thrips pupae and Fungus Gnat larvaeFeeds on Thrips pupae and Fungus Gnat larvae
BeneficialsBeneficials NematodesNematodesfeeds on Fungus Gnat larvaefeeds on Fungus Gnat larvae
Space RotationSpace Rotation It is critical to break the thrips lifecycle by It is critical to break the thrips lifecycle by
enforcing a chamber clean out every 3 months.enforcing a chamber clean out every 3 months.
All chambers in 005 Rightmire and in Biotech Support All chambers in 005 Rightmire and in Biotech Support Facility are common shared space. Facility are common shared space.
Researchers requiring a chamber with special temp or Researchers requiring a chamber with special temp or daylength for a specific project will need to pre-arrange daylength for a specific project will need to pre-arrange space with Gary.space with Gary.
There will no longer be any continuous cycling in There will no longer be any continuous cycling in chambers—all chambers will enter the rotation cycle.chambers—all chambers will enter the rotation cycle.
Space Rotation DetailSpace Rotation Detail
1 Planting Window ~6 weeks Maturation Window ~6-7 weeks
Cleanout
Planting Window
2 Planting Window Maturation Window
Cleanout
• Growth spaces will have a 6 week window for planting. This window may close sooner than 6 weeks IF space is full or INSV is detected. (Check for available space BEFORE planning to plant!)
• Spaces will have posted on them the last date for new plants. No new plants will be allowed in the space after the end date —even if there is space available.
• No plants are permitted to move from one space to another. • Once a space is empty, it will be cleaned by GH staff and heat
treated for 4 days, then reopened for planting.
Order of EntryOrder of Entry Order of entry is posted on the dry erase boards in the Order of entry is posted on the dry erase boards in the
BSF and Rightmire 005. Information is updated weekly BSF and Rightmire 005. Information is updated weekly based on actual pest counts.based on actual pest counts.
Users should note these daily as they go about their Users should note these daily as they go about their work to make sure they are following the correct order work to make sure they are following the correct order of entry.of entry.
Users who have entered a space lower on the list Users who have entered a space lower on the list cannot enter or re-enter a space above it on the list cannot enter or re-enter a space above it on the list until the next day.until the next day.
Dedicated lab coats are required for all growth spaces.Dedicated lab coats are required for all growth spaces.
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