How to Breed & Flask Phalaenopsis Orchids (Steps for Dry Seeds & Green Pod Propagation Methods) Tips on breeding orchids - at home!

In Flasking & Invitro Propagation
Scroll this

I flasked my first orchid seeds on November 7th, 2018, and while my preference is to avoid creating “exploratory content” and provide advice based on experience; in this case, it’s all I’ve got. I also found information about flasking orchids…sparse or not thorough. I plan on refining and updating this post and making it as valuable as possible so check back regularly. Once I’m a pro orchid breeder and flasker, I’ll make a video on my YouTube channel.

If you’re considering breeding orchids or flasking your own seeds, I hope you find this helpful. If you’re hesitant about flasking because you’ve never deflasked any orchid seedlings, I can tell you it’s not as tough as you may think. If you’re curious about the process, follow along in my deflasking orchid seedlings post where I deflasked my first set of phalaenopsis seedlings in September 2017.

This is a long post; you don’t have to read it all! These are the sections:

Video Slideshow: a quick flasking outlined later in this article

You might be wondering…

Q: Why do you need to flask orchid seeds? Why can’t we just plant them in a pot? They grow fine in the wild…right?

Answer: You need to grow orchids in a flask because orchid seeds don’t have the nutrient packs that other plants have to sustain the embryo. In nature, orchids form a symbiotic relationship with fungi called, mycorrhizae, which IS NOT the same fungus as a “mushroom”)—and through this relationship, the fungi provides sugar and necessary nutrients for the orchid…and even if there may be no (or very low) light for the orchid to photosynthesis, the fungus helps it grow and become a plant large enough to reach the light and then capable of sustaining it’s own life through photosynthesis.

Understand Orchid Biology & Seedling Development

Orchid Seed Development in Nature

In the wild an orchid will produce thousands of seeds from just one seed pod. The seeds are so light, they carry on a breeze and are dispersed everywhere; on leaves, on branches, on the trunk, on moss, and of course on the ground. Then the rains come and they pull the seed down into the crevasses and nooks, through cracks in the bark, through the faults of rocks, and into the moss – there, the seeds end up in dark and damp ideal place where both fungi thrive and where a tiny baby orchid will not dry out. The rest of the seeds generally do not make it past the germination stage because they just don’t land in the right place. Though the majority of wild seed will never become a plant, it’s a game of numbers which is why they produce so many seeds.

Unlike the seeds of regular plants, orchid seeds lack any form of nutrient storage. If you think of a pea or bean – the two chunky halves inside the skin…those are not the embryo itself but are in fact food for the embryo. Because orchids lack that nutrient reserve, they depend on fungi to survive.

Why we have to flask orchid seeds

Fortunately, scientists know it’s not the fungus specifically that’s required for an orchid to live, but rather it’s the nutrients a fungus provides the embryo. So with science we can look beyond fungi and understand exactly what the embryo requires and we can create those vital nutrients for developing orchids in a flask! THIS is why orchid seedlings must be flasked.

Bye mycorrhizae *polite wave*

 

Important Considerations & Roadblocks When Breeding Orchids

When breeding orchids, you will come up against challenges and considerations; this is important to work out early on.

BE DILIGENT – Label Everything

Do you know what’s a pain? Labeling everything. Do you know what’s even worse? Having a flower bloom 3 years later only to find out you had it labeled incorrectly and now don’t remember the original parents that you attempted at the time you made the cross. Another example, You’re flasking seeds in the lab, you’re doing multiple seed pods at a time, and then you get the flasks mixed up (because the seeds all look the same once they’re in a flask). You’re investing years of your time into this…take the extra effort to be diligent and label everything you do. Start a spreadsheet to track your hybrids. Record your attempts – the more info you track, the more you’ll know in the future about your successes and failures. And start a notebook so you can jot down items as you flask or are working so you don’t forget them…i’m terrible for “I’ll remember this later” (and then I put it off and forget all of the details).

Struggles of getting viable seed: Sterile plants, Difficult breeders, & Seed Infertility

When breeding orchids, something you should beware of is the relatively high occurrence of failure before seed is even produced; I would say of all the hybrids I’ve tried to make with my phals, I’ve only had about 8% success for pod creation – that’s like 1 pod for every 13 attempts (26 flowers burnt for one pod)! I work mostly with hybrids, so this is more of a common place issue, than with species, but here’s what you should know: the gene count of plants can affect it’s ability to produce seed (1n, 2n, 3n, 4n, etc); 1n (or “anuploid”) and 3n (triploid) plants are difficult to breed and/or can produce sterile seed (no embryo); sometimes too hybrids will create sterile offspring (like how a Mule is the sterile offspring of a horse and a donkey). This area of breeding is where it gets quite complicated; however, understand that you may experience high failure and relatively few seed pods simply because of genetics.

Should you breed hybrids or species orchids?

This is personal preference, but it comes down to what your goals are; do you want to make new hybrids or improve on old ones? or would you rather ensure that the purity of orchid species stays true? You can always do both; but it’s a consideration non-the-less.

Understand the difference between “sterilized” vs “disinfected”

Sterilized means there is no chance pathogens survived – heat is the way to sterilize. Disinfect is using chemicals to attempt to kill all pathogens…but sometimes they can survive. You sterilize the media and cutting tools, you disinfect the seeds, your gloves, and working space. It’s important to understand this distinction because you can see where potential contamination will happen.

Cleanliness is next to Godliness: Keep your workspace sterile and your seeds and seedpods disinfected

This is another challenge for me personally; you must keep your workspace absolutely disinfected. A single spore or bacterial cell (which is ALLLLL over your skin) can destroy all of your hard work. You need to treat yourself as the #1 vector, and you have to be SUUUUPER careful when moving things around. You’ll likely still get contamination, but how well you attend to this early on will determine how much success you experience.

Tips for keeping your work area sterilized and disinfected: Heat is the #1 way to kill pathogens. 121C is the benchmarked temp at 15minutes. That means 15psi on your pressure cooker and no lower. Fire (open flame) is a great way to kill viruses and kill anything on your tools.

Viruses start from the first cut of a seed pod – Flame all tools before use

Viruses are a different monster than bacterial and fungal infections…you don’t see them, they won’t show up in flask, and they can be transmitted to all of your seedlings simply by re-using a scalpel used to cut infected plant tissue. This means you can’t go slicing and dicing without flaming that blade between cuts. Why do you think viruses in orchids are so prevalent? It’s because tissue culture, stem propagation, is easy to cross contaminate if you don’t have absolute stringent sterilization techniques. A single unflamed scalpel can transfer a virus from one plant to all of your plants…including the clones. Same goes for seedlings – so be diligent.

Getting Started with Breeding: Timeline

Before we get into the processes of orchid breeding, I’d like to instill in your mind what type of commitment you’re making…

How long does it take for an orchid to go from flower, to seed, to flask, to a new flowering orchid?

The short answer is: A minimum of about 2.5 years; an average of 3-4 years; but up to 15 years in the case of some slow-growing Paphiopedilum.

The general stages of orchid orchid breeding & invitro flasking are:

  • Pollinating the flower, waiting until pod matures (~3-6 months for most phals but times vary by genera);
  • Preparing the media (1 day – including cooling time) – optional: wait 2 weeks to confirm no contamination;
  • Cutting the pod from the orchid,
  • Sterilizing and flasking clean seeds into a “Mother flask” (1-2 hours),
  • Waiting for seeds to grow and become protcorm like bodies (PLBs) (~4-12 weeks);
  • Optional: “Spread” replate #1 PLBs in new flask – only if germinated seeds are very dense & mounding on top of one another (1 day media, 2h flasking);
  • Replating #2 PLBs from mother flask into transplant (aka replate) flasks (1 day media, 2h flasking);
  • Wait for PLBs to form plantlets (~3-6 months);
  • Replate #3 – final transplant (1 day media, 2h flasking);
  • Wait for plantlets to grow into large seedlings (~3-6 months);
  • Deflasking (1-2h);
  • Growing seedlings to flowering-sized plants (~1-2.5 years).

 

Understanding what you’re getting into, if you’re still on board, let’s talk about orchid sex…

Step 1 – Pollinating an Orchid Flower – a Phalaenopsis

Before we skip right into pollination, there’s a quick lesson you need with regards to orchid pollen:

Orchid Pollinia

Unlike standard plants that have “powdery pollen”, orchids have cohesive masses of pollen known as pollinia. The pollinia is often hidden and many orchid have evolved specialized pollinators. Generally, the pollinia is stuck to a bug (the pollinator) with a sticky adhesive. Then the pollinator carries the pollen mass the next flower and pollination is achieved.

 

How to Pollinate a Phalaenopsis Orchid flower

Pollination Tools
  • Toothpick
  • Water – a very small amount works – in a shot glass
  • 2 flowers – one for pollinating, one for pollen
How to Pollinate an Orchid
Steps to Pollenating a Phalaenopsis Flower
  1. Grab a new toothpick
  2. Carefully pull the anther cap off the pollen parent and drop it onto a piece of paper or plate
  3. Carefully pull the cap off of the pollen – this is harder than it looks, and I frequently lose pollen that goes flinging off into the air
  4. Once the pollen is free from any stickies or bio mass, you want to transfer it to the female section of the flower
    *this must be done quickly or you risk dropping the pollen if the water dries:
  5. Dip toothpick into water (about 1/4″)
  6. With a light coating of water on the tooth pick, move it over a single pollen packet, and make light contact; the water tension should grab the pollinia
  7. Move diligently to the flower, and using your free hand, invert the flower upside down
  8. Position pollen into the flower and put the pollinia near the back of the flower – put slight amount of pressure to ensure it sticks to the sugary goop
  9. Slowly release flower to regular position
  10. Wait 24-48h and watch for the reception site to close over the pollen
  11. If the flower closes, and the pollen is accepted, the flower will begin to turn green or whither and the ovary will elongate. If the ovary dies, the pollination has not been accepted.

 

Photos of Successful vs. Failed Pollenations
Pollinated Flower Starting a Seed Pod

 

Failed phalaenopsis pollination attempts

Questions Regarding Flower Pollination

Should you use pollen from a flower you plan on pollinating (swap pollen for example)?

You can, I have, and I got a seed pod from the plant…that said, upon further reflection, I think it might be better to NOT use pollen from a flower you’re intending to pollinate, only because a flower will fade within 48 hours after it’s pollen has been removed. You probably want to give the plant maximum amount of time to close and deal with the pollen rather than jump-starting it into a fade.

Can I split or divide pollinia to fertilize more flowers from one flower? 

Yes; flame the blade of your scalpel before and after you cut the pollinia

Can I use multiple pollinia in a single flower?

Yes; but only do so if they’re from the same parent (don’t want a mix bag of babies) and, only do so if the pollinia is exceptionally small (example: Phal equestris)

How long does it take a pollinated orchid flower to mature into a pod?

Once a flower has been pollinated, you’re looking at about 4 months (120 days) to 8 months (240 days) for that flower pod to mature and produce viable seed. There is a document on green pod maturation times on the AOS website. But again, keep in mind individual species (and hybrids) may vary.

 

The Orchid Seed Pod

As the orchid seed pod forms, the ovary will swell as the embryos grow. The pod is full of chaff a white course, but fluffly looking material that helps push seed out of the pod once it’s ripe. Seeds nearest the column (where the flower was) will have a higher likelihood of being viable as the pathway from pollen to the eggs flow from the flower toward the stem. The image below (taken and observed by Allan Black) illustrates how the coloured seed hold embryos while the white seeds are sterile and have no viable embryo.

Photo of Orchid Seed Capsule
By Allan Black (Hobby Orchid Grower & Breeder)

Allan Reports:

Initially, I assumed the tan seed was either contaminated or some of the alcohol that I was using to sterilize the capsule had leaked into the seed (causing it to discolor to tan). I initially flasked the white seed thinking that it may be less likely to be contaminated or damaged. Later, I look at both color of seeds under a microscope and discovered that the white seed did not contain any plant embryos and that the tan seed contain lots of embryos. I re-sowed the flasks with the tan seed.Allan Black

 

Step 2 – Deciding the right method: Dry Seed vs. Green Pod

When it comes to sowing orchid seeds, you have two options: dry seed or green pod. A green pod means the pod is…still…green; and therefore the seeds are tightly bound within and still maturing. The thing about a green pod is, while the seeds are still maturing, you can flask them after about the half-way point. Dry seed, the other method, is when the seed pod has split open and the seeds are exposed to air…this means the seeds themselves are potentially contaminated with pathogens. Most people opt for the green pod method because there are fewer risks of contamination; however there are benefits and drawbacks to each method.

Why Flask Dry Orchid Seed

Advantages to Dry Seed Sowing
  • Long-term Storage & Future Practice; seeds can be stored for many years (though most report that viability declines quickly after pod “pops”) giving you a chance to try and try again if your first attempts fail.
  • Mature Seed; seeds are guaranteed to be mature (assuming the embryos are viable)
  • Easier; you put the seeds into a syringe and disinfect – then you blot a bit in each flask. For me this is easier than dealing with the green pod.
Disadvantages of Dry Orchid Seed Sowing
  • Greater risk of contamination b/c seeds are exposed to air before going to the media
  • Pod time (longer); takes on average 50-100% longer time to create a fully-ripe seed pod
  • Disinfecting process can kill seeds; if solution is too toxic, it kills the seed; if solution is not concentrated enough, pathogens invade.

Why Flask Green Pod

Advantages to Green Pod Orchid Seed Sowing
  • Lower risk of contamination; seeds are not exposed to bacteria or fungi provided the pod does not rupture and proper sterilization of the pod has taken place
  • Shorter maturation period; around 120 days vs. 220 for phals for example
Disadvantages to Green Pod Orchid Seed Sowing
  • One shot, no retries; if you have a contamination issue or your media is bad/wrong, you’re whole attempt is done.
  • Seeds may not be mature enough; some seeds take longer to mature and it depends on the species, not just the genera of orchid.
  • More “hands on”; if you’re working in a glove box, trying to fumble with and cut an orchid seed pod through foggy vision is incredibly difficult – to make matters worse, bleach makes the seed pod quite slippery so while it is supposed to be an easier method, I found it much more cumbersome than dry seed flasking.

 

Is it better to flask dry orchid seed, or a green pod?

Answer: Up to you! I wanted to try both methods and I prefer dry seed because it’s easier to get the seeds into onto the media. For you, the aspect of green pod may be more enticing because it has less chance for pathogens and is therefore overall more reliable for getting seedlings.

Here’s a video where I cover my justification for doing both dry seed and green pod: 

 

Understanding life-cycle of an orchid seedling

From Seed to Plant (and what’s in between – PLB protocorm like bodies), orchids go through 5 phases as they transition from an embryo to a “regular plant”; those stages are:

  1. [~80-250 days] Pollination & Seed Pod Maturation
  2. [2-3 weeks] Seed Sowing into Mother Flask, swelling of embryo, & “greening” of first PLBs
  3. [~3 months] Protocorm-Like Body (PLB) which is a green blob of tissue
    1. [~4 weeks] Small PLB (into spreading flask – if too many)
    2. [~6 weeks] Medium PLB
    3. [~2 weeks] Large PLB (into first replate)
  4. [~12th week] Differentiation – where the PLB establishes “up” vs “down” and roots and leaves begin
  5. [6-15 months] Vegetative Stage (into final replate)
  6. Transitional Stage (deflask)

 

Image of Orchid Development Stages in flask

 

Step 3 – Flasking Orchid Seeds

Okay, so you’ve gotten through all of the grit and you’re ready to flask, eh? Let’s dive in:

Checklist: What you will need to sow orchid seeds invitro

  • Orchid seed (a green pod or dry orchid seed)
  • Flasks and/or Vials
  • Orchid Seedling Media (Either Phytotech P668 – Orchid Maintenance Media with Charcoal or Sigma 6668 Orchid Maintenance Media)
  • A weigh scale that measures 0 – ~500g (I got one off Amazon for $20 CDN)
  • A contaminant-free workspace: either a glove box or a laminar flow hood (for those who have ample space).
    You can make a glove box yourself – I followed these great instructions to make mine; I would recommend getting a container with a gasket – the weather-tight storage boxes should be better for this.
  • A pressure cooker or autoclave (for those with loads of spare cash)
    • Note: The pressure cooker MUST go to 15PSI – At 15PSI, the temp is 121C – which will kill all bacteria and spores.
    • Beware: at 10PSI, you will risk contamination; that means you cannot use those fancy electric pressure cooker like your “Instant Pot”…PS: I tried this, it resulted in contamination. 
  • Chemicals for disinfecting your workspace & tools: chlorine or isopropyl alcohol
    • Household Concentrated Bleach (Sodium chloride): 10:2 (assuming bleach is 8.25% chlorine, this should be ~1.5% chlorine)
    • Isopropyl Alcohol 70% – be careful if you’re using flames to sterilize tools
  • Chemicals for disinfecting Green Seed Pods:
    • Concentrated household bleach: 10:2 (assuming bleach is 8.25% chlorine, this should be ~1.5% chlorine)
  • Chemicals for disinfecting Dry Seeds: either chlorine or hydrogen peroxide
    • Pool Bleach (Calcium chloride, CaCl2) or Household Bleach (Sodium chloride, NaCl)
      *For disinfecting your seeds: target .1–.5% chlorine
      **Reportedly Household bleach (NaCl) is more toxic to some seedlings than pool bleach (CaCl2); I have yet to explore the pool bleach, but I will.
    • Hydrogen Peroxide 3%
      *Use either bleach for disinfecting seed OR hydrogen peroxide…not both
  • Small amount of dish soap (works as a surfactant and ensures bleach/peroxide makes contact with the seed surface.
  • Syringes/plungers for seed sterilization (dry seed only)
  • Tools for cutting: scalpel/sharp knife (green pod only)
  • Tools for manipulating seeds: spoon, dental pick, or other small metal tools

 

Steps & Recipe for Making Orchid Flask Media

Mother Flask Ingredients

  • P668 – Phytotech Orchid Maintenance Media with Charcoal
    13.5 – 15g/L
    *I misread a recipe for replates and used 19g – oops
  • Sugar
    5g/L
    *I use coconut water – 5 g of sugar means 1/4 of a can that has 20g of total sugar
  • Gelling Agent
    3-5g/L
    *I used agar w/ a gel strength of 1.5% – 6g was not enough; 7.5-8 would have been better.
  • Distilled Water
    1L
    *this recipe makes a lot of flasks…probably around 15-20; consider halfing if you’re only doing one or two pods

Mother Flask Media Directions

Add agar to a small dish of cold water—this will make the process of dissolving it much easier. Bring water, sugar and gelling agent / agar-water to a rolling boil and turn off heat.

Pour 1/8-1/4” of media in each flask, put lid on top and put into the pressure cooker for sterilizing. Pressure cook at 15PSI for 15 minutes – up to 20 minutes.

Turn off pressure cooker and leave flasks in the pressure cooker until time to flask—if you try to rapidly cool the pressure cooker (in the sink with cold water), or if you open the seal too soon, you’ll risk contamination as the air rushing in to the pressure cooker brings pathogens.

Photo of Dry Phalaenopsis Seed Pod & Syringe 

 

Steps to sowing seed in mother flask (Flasking 1 of 3)

Assuming you have your glove box made (or a laminar flow hood) and all items from the checklist, you can proceed with the following steps:

  1. Prepare the Orchid Media, pour media into the flasks & vials, then sterilize at 15PSI for 15-20 mins.
  2. Allow Media to cool for 24 hours by leaving the pressure cooker and all contents undisturbed until ready for flasking (this will reduce contamination)
  3. Move all tools & sterilized orchid flasks into the glove box / working area
  4. Spray all items with bleach solution to sterilize the inside of the box; wait 15-20 minutes
  5. Spray again, and then proceed with work
  6. Sterilize seed or seed pod
    • Green Pod Instructions
      Video: Sowing Green Pod Orchid Seeds

      • Using bleach (Sodium chloride) – Goal concentration: .8-1.5% active chlorine
    • Dry Seed Instructions
      Video: Sowing Dry Orchid Seeds
      Tip: If you notice the pod yellowing, get a letter envelope and put it around the pod; the pod will split from the bottom, and the seeds will start to fall down – you’ll want to catch those (but keep them separate from the pod seeds, as there’s a higher chance these will be contaminated).

      • If using bleach (Sodium chloride) – Goal: .1-.5% active chlorine
        *coming soon*
      • If using pool chlorine -(Calcium chloride) – Goal: .1-.5% active chlorine
        *coming soon*
      • If using peroxide – 3%
        *coming soon*
  7. For each flask:
    1. After seeds or pod have been sterilized, transfer seed into flask
    2. Wipe inside of flask lid and mouth with 10-15% bleach solution (to prevent contamination)
    3. Place lid on flask
    4. Proceed to next flask
  8. When all flasks are complete
    1. Spray all flasks with bleach, then take them out one-by-one and;
    2. Place a thin strip of saran wrap around the top of the lid, covering the sides of the flask which will help prevent pathogens from getting in; alternately. Some orchid breeders use a larger single piece of tin foil and wrap it over the entire lid. I don’t like this because it blocks light from the top-down.
    3. Label each flask
  9. Place flasks near indirect light specifically avoiding any direct sun which may overheat the seedlings
  10. Wait 4 weeks (or longer) until you see protcorm like bodies
  11. Move to a brighter location, still avoiding direct sun
  12. If after 4 weeks the PLBs are mounding heavily, replate them into a “spread” of this same media recipe but in new flasks (simply spreading the seedlings out so they all get access to light);
  13. Wait 4-12 weeks until the PLBs swell heavily and start to differentiate (go from PLBs to plants w/ leaves);
  14. replate

 

[Immediately After Flasking]
Photos Sown Orchid Seedlings

 

[After 14-21 days]
Photos of the Flasked Orchid Seedlings
White bacterial contamination 🙁 – Phalaenopsis protocorms in flask
Bacterial contamination taking over Phalaenopsis protocorms 🙁
White fungal contamination in flask
Contamination-free test tube with phal seedling protocorms

 

After you’ve sown orchid seeds – caring for flasks

I honestly didn’t think this would be a thing I’d have to worry about. I figured that flasking would be the bulk of the effort, but as you can see, I started to get contamination in the large flasks after about 2-3 weeks. All of the little test tubes were contamination free while the big flasks all eventually got contaminated. After a bit of research here’s what I discovered:

Temperature Changes Can Cause Flask Contamination

I live in Canada; it’s winter; between day and night I have a swing from about 25C on warm days, down to 15C (lowest) on cool days. That 10C difference causes an expansion and contraction in the air within the flask. On my larger flasks (not threaded) air flowed more easily and this lead to contamination. All of my threaded (air-tight) test tubes were fine and yielded no contamination, AND none of my flasks that were waiting to be sown in the glove box experienced contamination either.

So what was the fix?

Orchid Seed Flask Incubation Chamber – Temperature Stabilization

I *just* started this experiment (last night on Dec 11th, 2018), so this is included for your info, but not as a recommendation—yet. I created an incubation chamber (something I used to do when I bred reptiles and needed to incubate their eggs). Using a Tupperware box, an aquarium heater, a bunch of water, and a cap full of bleach (to keep any pathogens from growing in the box), I created a temperature-controlled environment. I set the temperature to 87F/30C (in accordance with the AOS’s doc on orchid seed sowing where it mentions a “temperature controlled room”) and placed a few flasks inside. I am concerned about how humid it is inside the box…but we’ll see how it goes.

Photos of the Orchid Flask and Seedling Incubation Chamber

As a plan B – my buddy Dan has noted that he uses pink styrofoam and a reptile heat pad to do the same job. As long as the temp from the pad is consistent and there are no breezes, in theory this should also keep the temp inside the flask consistent and prevent that ebb and flow of air as the temps outside change.

I will update this if I experience success…or remove it all together if it results in disaster.

Spreading: Taking PLBS from one mother flask into new mother flask
(Optional, but best done if seedling density is high)

This is a step is only required if your protocorms are piling all over one another. What will happen if you don’t is the seedlings under the top layer will eventually die. Spreading them into a new mother flask allows both flasks the opportunity to feed ALL the seedlings. It’s always a risk for introducing contamination, so ideally you want to sow less seeds and avoid this step if possible.

Steps for Spreading Orchid Protocorm-like Bodies

Take the mother flask and a second un-sown flask with mother media and put them into the glovebox. disinfect your work area and outside of both flasks. Use a tool (spatula or whatever) to scoop PLBs from one flask into the second. Do your best not to touch anything other than the media or PLBs. Spread them in the new flask and redistribute the PLBs in the old flask. Your goal is to end up with two mother flask.

Photos of Spreading Orchid Protocorm-like Bodies

 

Light/No Light (phals vs… paphs) – “largely unknown – will do more research; but understand that light plays a role”
*coming soon*

Replating from the Mother Flask (Flasking 2 of 3)

Replate Flask Ingredients (for Protocorm Like Bodies (PLB) Transplant)

  • P668 – Phytotech Orchid Maintenance Media with Charcoal
    20g/L
  • Sugar
    5g/L
  • Banana *better if lightly green
    25g/L
  • Potato – cooked (can be microwaved) then mashed
    50g/L
  • Gelling Agent (or 8g agar)
    3-5g/L
  • Distilled Water
    950ml/L

 

Orchid Replate Instructions – Blend Potato and Banana with some distilled water. Like with the mother flask, add the agar to cold water, then mix all ingredients together. Check the pH – ideally the pH should be around 5.5-5.8.  The P668 media is usually adjusted, but adding banana and potato can alter the desired pH. Adjust the pH if needed (citric acid for more acidity, baking soda to increase alkalinity). Once that is done bring the mix to boil. Then fill the flasks (1/4” deep) and autoclaved.

Glovebox setup: to begin work in the glovebox, make sure you have these items:

  1. A small dish of peroxide (3%) with less than a drop of dish soap
  2. A small dish of bleach (25%)
  3. A spray bottle with bleach (20%)
  4. Tweasers and/or metal tools that have been sterilized with fire before starting work.

Once flasks are sterilized and cooled, place them into the glovebox with the mother flask. Spray the outside of all flasks and tools with the bleach solution (20% bleach, water, & a drop of dish soap). This step is important for preventing contamination of both your replates and your mother flask, so be diligent and liberally spray the flasks paying special attention to the lid and rim. Let everything sit wet with the bleach water for 10 mins, the proceed with transferring the protocorms to the replate media.

For best results: after each tool is used for a replate, put it into the bleach solution for 30 seconds, then transfer it to the peroxide solution to rinse off the bleach. Then proceed with the next replate. Once all the replates are completed, leave them under lights for the next 12 hours which will neutralize any peroxide. As an aside: the purpose of the peroxide in this case isn’t for sterilization, but it ensure that you’re able to rinse the bleach off without re-infecting the tool if you were to use a bowl of water.

 

Final Replating of Orchid Protocorm-Like Bodies (PLBs) – when, why, how (Flasking 3 of 3)

Final Replate Flask Ingredients (for Protocorm Like Bodies (PLB) Transplant)

  • P668 – Phytotech Orchid Maintenance Media with Charcoal
    25g/L
  • Sugar
    5g/L
  • Banana *better if lightly green
    25g/L
  • Potato – cooked (can be microwaved) then mashed
    75g/L
  • Gelling Agent (or 8g agar)
    3-5g/L
  • Distilled Water
    950ml/L

 

Orchid Replate Instructions – Blend Potato and Banana with some distilled water. Like with the mother flask, add the agar to cold water, then mix all ingredients together. Check the pH – ideally the pH should be around 5.5-5.8.  The P668 media is usually adjusted, but adding banana and potato can alter the desired pH. Adjust the pH if needed (citric acid for more acidity, baking soda to increase alkalinity). Once that is done bring the mix to boil. Then fill the flasks (1/4” deep) and autoclaved.

*coming soon*

 

Deflasking Orchid Seedlings

So you’ve made it this far eh? I created a similar type post for my first deflasking – you can follow along in this other article.

Information & Resources for Orchid Breeding, Seed Flasking, & Media

 

Special Thanks

This post was a culmination of research and advice from my orchid friends around the world. I’d like to give a thanks to Adeljean Ho – part owner of Neblina Orchids, Ken Avent – renowned orchid breeder, John Barnes, Pat – from Kingfisher Orchids, and the groupies over at the International Phalaenopsis Alliance (IPA) – including: Robert Bedard, CJ Mack, Ben Belton and others.