all rights reserved © 2018borneo exotics (private) limited

All Rights Reserved © 2018 Borneo Exotics (Private) Limited

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Exclusive guidance for Borneo Exotics’ Customers

Introduction Establishing plantlets from tissue culture at home as well as the professionals can, is not difficult, provided a few simple rules are followed. When you receive your plantlets, they will have been growing on a sterile jelly (agar) that contains all the nutrients they need and will have never experienced dry air, or much change in temperature, or fluctuation in light levels. The process of establishing them is known as “weaning” and it’s exactly that: they need to be gradually weaned, or acclimatized, to normal growing conditions, where they will then have to withstand the presence of pathogens (bacteria and fungal spores which are everywhere), varying humidity, temperatures and light levels. This guide shows several possible cheap and easy options to achieve a successful result, but there are other ways too. You can use your ingenuity and available materials to modify the methods we describe. But whatever you do, the points in the next section need to be followed:

The Basics Before we get into the specifics, we’ll firstly give you the basic do’s and don’ts you need to bear in mind. Let’s start with the cautions:

Slowly does it! Any changes in conditions must be made slowly! This is key to success. Sudden changes in conditions may cause stress, or shock to your plantlets.

Humidity The plantlets you receive will have never known anything other than 100% humidity. Until they are weaned, if you leave them in dry air for even a moment, they will wilt, and you may lose them. As you remove them from their flasks, you need to put them straight into water at room temperature. Then they need to be kept completely wet always, until they are transferred into their humid weaning chamber as described later. No Direct Sun!!! Please never, expose your plantlets to direct sunlight until they are fully weaned and even then, do so slowly over a period of time. When they are unweaned, if you leave them in the sun, even in their original flasks, it will take only a couple of minutes to kill them. The greenhouse effect will cause a rapid heat build-up inside the flasks. This also applies to the weaning chamber you are going to construct. The best and easiest way to handle and grow the plantlets is under LED or fluorescent lights.

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Weaning Guide


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Factors for Successful Weaning Light Light levels are not critical at the outset but become more important as the plantlets establish and grow. We have experimented for many years and find that cheap LED desk lights about 20cm (8”) above the plantlets work extremely well. The type of light, or whether it’s warm-white or daylight doesn’t seem to matter. For example, the cheapest of the IKEA desk lights work just fine, and there is no need to buy expensive “grow lights”. In their natural habitat, the plants would have a 12-hour light-dark cycle, but our trials have shown that that for weaning, if it’s easier, you can leave the light on continuously and it doesn’t seem to make any difference. Humidity& Roots To start with, you will need to keep your plantlets in some sort of enclosure or “weaning chamber” to keep the humidity at close to 100%. This is because the roots that form in tissue culture do not work when the plantlet is no longer in sterile conditions. They will disappear, and new roots will have to form. Until that happens, the plantlets cannot uptake water. The plantlets you receive may not have roots at all, because they really aren’t necessary to start with. What is important, is that even if no roots are evident, there should be a little black area on the base of the plantlet. This is known as “callus tissue” and it’s from this that the new roots will emerge. In effect your plantlets are like cuttings, except that they already “know” that they need to form roots from this point. You should not need to use any rooting hormone.

Pathogens Pathogens are germs or fungi which are always in the environment. Just as with animals, plants have an immune system, giving them a natural defense against pathogens. But your new plantlets have been in sterile culture all their lives and will need to build up their immunity. They will also be stressed from the transport, which makes them more susceptible. This isn’t a problem, provided you observe the following precautions:

• Handle the plantlets very carefully, using clean hands, tweezers and scissors. We use a dilute chlorine solution (bleach at 5%) to sterilize instruments, but other alternative sterilizing


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agents may be used. Be sure that whatever chemicals you use are not poisonous to plantlets (or you)! If you don’t disinfect, then at the very least, make sure instruments are thoroughly washed before use and re-wash them between plantlet batches.

• If you can, we recommend you use a precautionary fungicidal dip just prior to planting out. This isn’t completely essential, but it may increase your overall success rate. We dip in a relatively harmless systemic fungicide: thiophanate methyl. It’s sold as ‘Topsin’ in some parts of the world and ‘Cleary’s 3336’ in the USA. Use at the manufacturer’s recommended dilution and observe all recommended safety precautions. There’s no need to wash the fungicide off the plantlet after dipping. If this fungicide isn’t available where you are, then ‘Captan’ is useful as well as relatively safe to handle. There are also others.

Temperature This becomes more critical after weaning, but general, lowland Nepenthes need warm conditions, between 24 deg C and 36 deg C, intermediate Nepenthes need temperatures of between 15 deg C and 26 deg C and highland Nepenthes need temperatures of between 12 deg C and 22 deg C (or lower for some highland varieties). These are all approximate temperature ranges; the web and the many Nepenthes forums are a great source of further information. Growing Media There are so many types of media used to grow Nepenthes that it’s beyond the scope of this guide to discuss them in any detail. A search of the web will provide a lot of information, but for weaning plantlets, we will recommend two which are popular:

• Sphagnum moss, preferably live, but dead moss still works. • Peat-moss.

Whatever media is used, it needs to be always wet and naturally acidic.

Nutrients Your plantlets will arrive already pumped-up with nutrients and we don’t recommend using any fertilizers during the weaning process. Once weaned after about 10 weeks, the plantlets will benefit from being treated with a foliar fertilizer. If you’re new to growing Nepenthes, there’s plenty of information available on the web. But don’t fall into the trap of thinking that just because these plantlets are carnivorous, that they don’t benefit from fertilizing – they do!

The Weaning Process For the first 7 weeks you need to try to keep the conditions as constant as possible while the plantlets establish, grow new roots and their first 1 or 2 new leaves. Please resist the temptation to open the weaning chamber to look at the plantlets too often, and never change anything suddenly. Because the weaning chamber should be sealed, there’s no need for watering, as everything should be wet at the outset and naturally stay that way. After about 7 weeks, you can gradually start to expose the plantlets to lower humidity until they are fully acclimatized to the outside conditions, which may take about another 3 weeks. We describe alternative ways this can be done in a later section.

Preparation Before your shipment of plantlets even arrives, it’s wise to get everything ready in advance. You will need to prepare:


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A suitable weaning chamber as described in the next section. A fungicide (optional) as described in the section on ‘Pathogens’ above. Tweezers (preferably the soft type sometimes used for contact lenses). Small scissors for trimming off dead leaves. Containers for soaking and washing the plantlets. Labels pre-marked and ready. Remember that most tissue culture plantlets look identical to

each other, so do be careful to keep your labels in order!

When your Plantlets Arrive Inspection Before opening any of the flasks, check them all carefully. Once the security seal is broken, any warranty is void. The plantlets will be mixed up with the agar due to shaking in transit, but you should be able to see that they are green and alive. Remember to do all handling of the flasks and plantlets out of direct sunlight.

typical tub of plantlets at the time of dispatch typical tub of plantlets after shipment

The tubs you receive are likely to look like the photo on the right, which is normal. The agar will have liquefied in transit due to vibration, but the plantlets should still appear green. If the plantlets are brown or a dark khaki-green, please do not open the tub and refer to our Replacement Warranty under the Ordering Guide and Terms& Conditions.

Opening the Tubs & Washing the Plantlets Deal with each tub one at a time. Immediately a tub is opened, the plantlets need to be removed and soaked in water. After checking that the plantlets in each tub look OK, open them and remove the plantlets. You should have already prepared suitable containers of clean water at room

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http://borneoexotics.net/docs/retail_TC.pdf


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temperature and labels for each variety. The volume of water should be sufficient for each batch of plantlets (at least ½L or 1 pint per batch) and must be changed between batches. Soak each batch of plantlets for 1 hour in the first container of water and for a further 15 minutes in another container of water, to remove all traces of agar. If any agar remains on the plantlets, it may cause rot. Take the opportunity to trim off any dead or damaged portions of leaves.

typical tub with lid removed typical (large size) plantlet pre-washing

We highly recommend pre-preparing your labels with all relevant information for your inventory system before commencing the planting-out process. To try to do this as you go along, is to risk mixing-up plantlets. At the tissue-culture stage, nearly all Nepenthes look identical – so beware! The safest way is to drop the label in with each batch of plantlets when you wash them and ensure the label stays with those plantlets right the way through until they are planted out and installed in their weaning chamber. After the second wash, soak each plantlet in the fungicide preparation for 10 minutes (if you have a suitable fungicide). As discussed in the section on Pathogens it’s not essential to use fungicide, but it possibly helps.

Planting-out Immediately after washing each batch of plantlets, plant them out straight from the fungicide soak. There’s no need to wash the fungicide off afterwards. If the plantlets appear to have no roots, don’t worry -as explained earlier, they aren’t necessary at the outset, since new ones need to form anyway. However, do gently firm each plantlet in, so that the media is in contact with the base of all the plantlets. The photos below show the procedure of planting out into a peat-moss media. During the entire planting-out process it’s essential to keep the plantlets wet by regular spraying.


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carefully prick out and firm in each plantlet

The tray or pot should then be immediately placed into the weaning chamber which you have already prepared.

The Weaning Process There are very many ways to achieve conditions of 100% humidity. Here we show 3 suggested methods, all of which we have tested and all of which work well. You need to have prepared the materials for this before the shipment arrives and certainly before you start planting-out. For all these methods, you can provide light during the weaning process using a desk LED reading lamp placed about 15cm (8in) above the level of the top of the plantlets. The strength of the lamp, or whether it’s warm-white or daylight doesn’t much matter. For example, the cheapest IKEA desk lamp works perfectly well. There’s no need to water the plantlets during the weaning process, just ensure that everything is very wet to begin with when the weaning chamber is sealed.

Method 1 - Plastic Bottle Take a plastic drink bottle large enough for the pot you’re using (here we are using a 1.5L bottle) and cut it into two halves. Place your pot in the lower section and you should be able to then push the top section over the bottom section as shown, sealing the unit entirely.

first 7 weeks – fully enclosed 8th week – cap removed 9th - 10thweeks- top half removed


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Leave to stand in LED light for 7 weeks and then remove the cap. After another week, remove the top half of the bottle as shown above. After another 2 weeks, the pot can be removed entirely from the bottle, as weaning will then be complete. Method 2 – Tub with Polythene Cover Take a clear plastic tub large enough for the pot you’re using. Be sure to leave enough headroom for the plantlets to grow during weaning. Place your pot of plantlets into the tub and fix a piece of polythene over the top using either an elastic band, or tape, to fully enclose the plantlets, then follow the steps shown below. After the 10th week, weaning will be complete.

first 6 weeks – keep fully enclosed 7thweek – cut small holes in the polythene

8thweek – cut larger holes in the polythene 9th - 10thweeks - remove polythene cover

Method 3 – Plastic Bag Place the pot inside a clear plastic bag, seal the top using an elastic band and follow the steps below.

first 7 weeks – keep fully enclosed 8th week – open bag and extend upwards 9th - 10th weeks – gradually roll bag down


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Here are 3 different Nepenthes varieties after 10 weeks of weaning using the plastic bag method:

3 different varieties after 10 weeks of weaning

Post-weaning Whichever method you use, the plantlets should have formed new roots by 10 weeks, as well 1 or 2 new leaves. They are then ready to be able to withstand outside conditions which are suitable for nursery-grown Nepenthes. Except for a few carefully bred hybrids, this doesn’t mean indoors in very low humidity, but in suitable temperature and humidity for the varieties you are growing. Fertilizing By the time the weaning process is complete, the plantlets will need feeding or growth will slow. We recommend using a fortnightly foliar spray of 20-20-20 or almost any household fertilizer or orchid feed at recommended dilution. Again, the web is a vast source of information for which types of Nepenthes benefit from which fertilizers. Robert Cantley Sri Lanka 20 Aug 2018

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