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Thread: Introduction of Nepenthes into Tissue Culture from Vegetative Tissue

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    Introduction of Nepenthes into Tissue Culture from Vegetative Tissue

    I've seen in various threads the question of introducing Nepenthes into TC via vegetative tissue (meristematic (apical or nodal)) from time to time. It sounds like it has been a difficult avenue for people, resulting in the fact that most, if not all, commercial TC production of Nepenthes comes from seed introduction and subsequent micropropagation. I would like to know if there has been any progress in this aspect of Nepenthes TC. If not, is there any demand for it? Is the current supply of TC produced Nepenthes, originating from seeds, adequately addressing the current demand for Nepenthes? The most obvious advantage of TC production derived from vegetative tissue is the mass propagation of a hybrid or species line that carries desirable traits that may be lost or modified if reproduced through seed production (leaving aside discussion of somaclonal variation).

    I have been debating recently whether or not to get back into doing TC. I have a degree in horticulture and have many years of experience doing TC. I used to work for a nursery that specialized in the introduction of new perennial plant cultivars to the US market. I had successfully introduced, from vegetative tissue, various plant species such as Hosta sp., Heuchera sp., Tiarella sp., Silene sp., Corydalis sp., x Heucherella sp., Brunnera sp., Dionaea m., D. capensis, Nicotiana sp., and others that I cannot remember (it has been over 15 yrs). At the nursery I worked for, I would be responsible for coming up with the protocols for introducing (stage I), multiplying (stage II), and rooting (stage III) the novel plant. We would then send the protocol and plant off to another TC lab for mass production. After working in that environment 40 hrs/week for several years, I ended up developing carpal tunnel and had to stop. Now, I work for a biotech company in the Bay Area. Since I haven't been doing TC for a number of years, my carpal tunnel has gone away. During my time doing TC, my contamination rate was very low, even when dealing with starting materials such as a Hosta where you are digging it up from the ground to get access to the growing bud. I did not rely on antibiotics or PPM, just good techniques and a dissecting microscope.

    I've recently got back into growing CPs. I noticed that there has been some discussion of Nepenthes TC here and there but little as of late. Before I embark on a potentially labor intensive project, I want to know from other CPers if its going to be worth my time. I have access to all the necessary equipments with my current job (flow hood, autoclave, scalpels, forceps, dissecting microscope, media, PGRs, etc.). What I lack is an incentive to move forward. If I were to start, I currently have N. x ventrata to play around with. Is this a particular easy plant to get vegetative tissue into culture without contamination? If so, I would need to find something more challenging. However, I might be getting a bit ahead of myself. First thing first, is there a need/desire?

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    Grey Moss's Avatar
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    TC using meristematic tissue of nepenthes has been done now, by at least 1 nursery in the US. It reportedly had low sucess rates, but it has now proven to be possible. I'd say there is certainly a demand for it. Lots of desirable cultivars and other individual nepenthes would definitely benefit from being more widely available.

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    Quote Originally Posted by Grey Moss View Post
    TC using meristematic tissue of nepenthes has been done now, by at least 1 nursery in the US. It reportedly had low sucess rates, but it has now proven to be possible. I'd say there is certainly a demand for it. Lots of desirable cultivars and other individual nepenthes would definitely benefit from being more widely available.
    Thanks for the info. Do you happen to know if the low success rate is due to high contamination rates, poor multiplication, or poor growth in TC?

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    Quote Originally Posted by WeR PlantFood View Post
    Thanks for the info. Do you happen to know if the low success rate is due to high contamination rates, poor multiplication, or poor growth in TC?
    It is a bit of all of the above. I have had some success in introducing Nepenthes from cuttings or from basals. They usually involve extreme steps in terms of sterilization, almost to the point of killing the explants; skillful dissection of the plant matetial; the use of antifungals and, often, antibiotics. Progress is generally slow and rescue cultures are the norm.

    It is far more economical to produce them from seed . . .
    “Sì perché l'autorità dell'opinione di mille nelle scienze non val per una scintilla di ragione di un solo . . ."

    -- Galileo "Biff" Galilei

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    Quote Originally Posted by BigBella View Post
    It is a bit of all of the above. I have had some success in introducing Nepenthes from cuttings or from basals. They usually involve extreme steps in terms of sterilization, almost to the point of killing the explants; skillful dissection of the plant matetial; the use of antifungals and, often, antibiotics. Progress is generally slow and rescue cultures are the norm.

    It is far more economical to produce them from seed . . .
    Hi BigBella! Thank you for your response. I've read through many of the threads on Nepenthes TC and noticed that you are one of the few individuals at the forefront of this topic and hope that I might be able to pick your brain a bit. Have you found very different media growth conditions for different species of Nepenthes or have they been fairly homogeneous? Do you still use PPM or have you moved away from it? I was an early user of PPM back in '99 when Carol Stiff sent me a sample to try (she was my prof in college). I was never really convince in its use and did not adopt it for general TC use.

    Although I don't doubt that it is far more economical to produce them from seed (one reason for my original post), it intrigues me to find out if my experience with TC combined with my experience in biotech (dealing with plant transformation, protein purification, and bioassays) can overcome some of those difficulties associated with Nepenthes TC. Setting that aside, is it your opinion that the current demand for Nepenthes is being met by the current method of TC via seed introduction? How is the industry dealing with the fact that seed derived TC plants will have undefined characteristics until after the plants are grown up? Do they just maintain a small batch of culture and quickly send out a few plants for ex vitro growth? Do they just discard into the trash or dump into the market a poorly performing clone (bad coloration or growth characteristics)? At the nursery I worked for, the plants from their breeding program that goes into culture always get evaluated ex vitro first. This process often takes years before a plant is selected to go into TC for mass production because the plant is evaluated for growth and culture characteristics in addition to phenotype. It seems that the current Nepenthes TC seems a bit backwards to me. However, if the wheel ain't broke, why fix it?

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    BigBella's Avatar
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    Quote Originally Posted by WeR PlantFood View Post

    Setting that aside, is it your opinion that the current demand for Nepenthes is being met by the current method of TC via seed introduction? How is the industry dealing with the fact that seed derived TC plants will have undefined characteristics until after the plants are grown up? Do they just maintain a small batch of culture and quickly send out a few plants for ex vitro growth? Do they just discard into the trash or dump into the market a poorly performing clone (bad coloration or growth characteristics)? At the nursery I worked for, the plants from their breeding program that goes into culture always get evaluated ex vitro first. This process often takes years before a plant is selected to go into TC for mass production because the plant is evaluated for growth and culture characteristics in addition to phenotype. It seems that the current Nepenthes TC seems a bit backwards to me. However, if the wheel ain't broke, why fix it?
    Nepenthes are a bit like any other ornamental plant; and strains are chosen for faster growth; pleasing coloration; and general vigor.

    Typically, the fastest growing plants in TC are most likely to reach market. They are quickly multiplied, while some other lesser cultures are either stored or even destroyed. Nowadays, multiple clones of more popular species are available.

    Most nurseries aren't comfortable with the notion of waiting upwards of ten years to get a flowering sized potentially fugly highlander; and they'd rather enjoy the current demand for plants just ex vitro . . .
    “Sì perché l'autorità dell'opinione di mille nelle scienze non val per una scintilla di ragione di un solo . . ."

    -- Galileo "Biff" Galilei

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    Quote Originally Posted by BigBella View Post
    Nepenthes are a bit like any other ornamental plant; and strains are chosen for faster growth; pleasing coloration; and general vigor.
    I guess there is a bit of an assumption that the characteristics for that particular stain is stable and lightly to be pass on to its progeny. Although this is not guaranteed coming from sexual reproduction.


    Most nurseries aren't comfortable with the notion of waiting upwards of ten years to get a flowering sized potentially fugly highlander; and they'd rather enjoy the current demand for plants just ex vitro . . .
    I find it a little funny considering all the criticism the nursery I worked for got for trialing their plants too quickly (2-4 years vs. the more standard 4-5 yrs). With that said, it is easy to understand the economics behind not waiting upwards of ten yrs to sell a plant. However, it is not always necessary to trial a plant to the flowering stage as long as the characteristics marketed is stable and reproducible. I guess consider us lucky that we're not dealing with trees (fruit or ornamental) where the trial periods could easily be a decade or two.
    Last edited by WeR PlantFood; 09-04-2018 at 06:12 PM.

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