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Best Practices Best Practices For researchers using For researchers using Biotech Support Facility & Biotech Support Facility & Rightmire Hall Growth Chambers Rightmire Hall Growth Chambers and Greenhouse and Greenhouse

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Best Practices. For researchers using Biotech Support Facility & Rightmire Hall Growth Chambers and Greenhouse. Introductions. Gary Posey, Greenhouse Superintendent Day to Day at BSF & Rightmire posey.8 292-9241 Joan Leonard, Program Manager Day to Day at Bio Sci Greenhouse - PowerPoint PPT Presentation

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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 success

3.3. Improve uniformity and repeatabilityImprove uniformity and repeatability

4.4. 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 Overwatering: #1 cause of plant diseases, pest

problems and plant death! Too much-Too often.problems 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 Green algae growing on soil surface is sure indication of overwatering.of 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. cucumeris

Feeds 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

NematodesNematodes

feeds 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.