hey everyone,
I'm writing a book! Well, 2, but the one I want to talk about here is a book about my perception of the world and its life. I am writing about 100 (or more) species I've met along my journey here, and how I feel about each one of them. I do not know when it will be finished, but I wanted to share a few paragraphs from the plant section
Kingdom Plantae
I love plants. There are a few specific groups I’m especially keen on, and we’ll get to those soon enough. But plants in general are also a passion. I need to know my plants if I’m going to rear moths successfully. Please note for this section that plant taxonomy is different from animal taxonomy, which I’m more familiar with. Plants have less clear distinctions between phyla and classes, and so the taxonomic rankings in this section need to be taken with a few grains of salt.
Phylum Tracheophyta
The phylum Tracheophyta is the phylum of vascular plants. Plants are initially split from their ancestors the algae by being necessarily multicellular, specialized (with tissue types and sometimes organs), and by having a lining of sterile (empty) cells around the gametes. This last characteristic is something no alga except for the plants has. Plants are typically said to be vascular or non-vascular. Non-vascular plants include mosses and hornworts, and these plants are very small and can only exist in very wet areas. We’re going to skip the non-vascular species here. It’s also important to note that plants don’t necessarily have to be autotrophic to be considered plants, although autotrophy is the ancestral state.
Class Lycophyta
The lycophytes are our first group of vascular plants. Without true leaves, they are limited in their size, but not in their mystic qualities.
Order Lycopodiales
Family Lycopodiaceae
This is a family that fascinates me. I didn’t learn about it until a few years ago, actually. They are called “princess pine” at least in New England and used to be harvested excessively for wreath making. Some species are endangered or rare as a result of this. What’s surprising about this family is that the spores of these plants (very much unlike typical/true fern spores) require a mycorrhizal symbiont to germinate. Much like orchid seeds (which we’ll get to later), these spores can be induced to grow on a tissue culture medium. Enter Dr. Dean Whittier. He was the one who discovered you could use TC medium to germinate the spores of these plants. Over a decade later, Brian Aikins of American Fern Society sends me spores of 4 Lycopodiaceae species. I attempted to get spores to germinate on 3 different formulations. One was a Knudson medium, one was called Parker Thompson Fern medium, and one was called Murashige fern medium. I poured plates of each medium. I took the tiny spores of each species and subjected them to a vacuum treatment to force water into them, a 10-minute bleach bath followed by a rinse with sterile water, and being plated. The vacuum treatment is a really cool process btw. It works by using water with a surfactant, which is a fancy term for something that resists floating by objects. Add a bit of detergent to some water and add spores. Then, swirl the solution and subject to vacuum pressure. Air is evacuated from the vessel and water enters its void, including the microscopic voids in the spore surface. This causes the spore to take up water, swell, and sink. This is CRUCIAL for surface-sterilizing the spore. Remember that tissue culture must be done in a sterile environment, and unbleached fern spores are anything but sterile.
So I then took my imbibed spores and put them in a solution of 10% bleach and a little surfactant to make sure the bleach really got onto that spore. I let the spores soak for 10 full minutes. I filled the bleaching vial to the brim, and then I surface sterilized the capped vial and put it within the flow hood. I shook it back and forth a few times to resuspend the spores. I have discovered you can make your own sterile rinse station for the next step. Get a pint mason jar. Fold a coffee filter in fourths and stuff it in the jar and cap it. Put the jar ring on loosely and autoclave/pressure cook for about 1 hour. Tighten the jar ring and then, when you must use it, you just pull out the coffee filter, make it into a cup, fit it over the rim and that sterilized ring will hold it in place! The air inside of the jar is also sterile (and yes it matters). I poured the bleach with spores into the filter after the appropriate time and rinsed them with sterile water. Then with a sterile pipette I dripped some spore water over the media and swirled the plates before sealing with parafilm, labeling, and storing in the dark.
Let’s pause with this story line and introduce another one. Whittier and his work, to my knowledge, didn’t ever see sporophytes forming in the species I’m currently working on. I wonder if using the medium I had success on (which I will reveal soon) but with different amounts of glucose or more nitrogen or something would help. I, at present, have yet to see sporophytes. I have more spores, and I plan on doing experiments with different concentrations of different things (the glucose concentration of the medium I experimented with earlier was 0.5%, or 5 grams of glucose per liter of medium) such as glucose and nitrogen to see if that influences germination rate, time, or sporophyte formation. I’ve read that similar species’ spores germinate better at a low glucose concentration but the subsequent gametophytes grow more rapidly at a higher concentration. Also, glucose is better for these gametophytes than sucrose is. I do not, however, know what is required for the formation of a sporophyte.
It took about a year. I kept the petri plates without any germination for an additional year. But 11 months after I sowed the spores, I noticed a few tiny whitish blobs on 1 medium. They were maybe the size of a grain of salt. I reckon each plate had 10,000 spores and only a handful of gametophytes showed up. Which basically means there was a 0.01% germination rate. Ok maybe more like 0.05%. The only medium with any germination after 2 years was the Parker-Thompson fern medium. A few months later, I put some of the cultures in the light and kept some in the dark. This was because I suspected that light would trigger the gametogenesis necessary for sporophyte formation. So this section will be dedicated to those 4 species. What did I observe with them? Did any make it to sporophyte? Hehe let’s see!
Dendrolycopodium dendroideum
I just wanna say that I have this species near my childhood home and I’d love to take some spores of it one day. I did get spores from Brian Aikins so I will have things to report on from this species.
Lycopodium clavatum
I almost got shot over this species. Why? Well because I was under the impression that we NEEDED a member of this genus for a teaching lab at URI. I had sections of D. dendroideum, which had formerly been classified as a Lycopodium. Not knowing this, I wandered into a shooting range where I found a clump of actual Lycopodium. I have gametophytes of this species at present.
Diphasiastrum digitatum
Diphasiastrum tristachyum
I'm writing a book! Well, 2, but the one I want to talk about here is a book about my perception of the world and its life. I am writing about 100 (or more) species I've met along my journey here, and how I feel about each one of them. I do not know when it will be finished, but I wanted to share a few paragraphs from the plant section
Kingdom Plantae
I love plants. There are a few specific groups I’m especially keen on, and we’ll get to those soon enough. But plants in general are also a passion. I need to know my plants if I’m going to rear moths successfully. Please note for this section that plant taxonomy is different from animal taxonomy, which I’m more familiar with. Plants have less clear distinctions between phyla and classes, and so the taxonomic rankings in this section need to be taken with a few grains of salt.
Phylum Tracheophyta
The phylum Tracheophyta is the phylum of vascular plants. Plants are initially split from their ancestors the algae by being necessarily multicellular, specialized (with tissue types and sometimes organs), and by having a lining of sterile (empty) cells around the gametes. This last characteristic is something no alga except for the plants has. Plants are typically said to be vascular or non-vascular. Non-vascular plants include mosses and hornworts, and these plants are very small and can only exist in very wet areas. We’re going to skip the non-vascular species here. It’s also important to note that plants don’t necessarily have to be autotrophic to be considered plants, although autotrophy is the ancestral state.
Class Lycophyta
The lycophytes are our first group of vascular plants. Without true leaves, they are limited in their size, but not in their mystic qualities.
Order Lycopodiales
Family Lycopodiaceae
This is a family that fascinates me. I didn’t learn about it until a few years ago, actually. They are called “princess pine” at least in New England and used to be harvested excessively for wreath making. Some species are endangered or rare as a result of this. What’s surprising about this family is that the spores of these plants (very much unlike typical/true fern spores) require a mycorrhizal symbiont to germinate. Much like orchid seeds (which we’ll get to later), these spores can be induced to grow on a tissue culture medium. Enter Dr. Dean Whittier. He was the one who discovered you could use TC medium to germinate the spores of these plants. Over a decade later, Brian Aikins of American Fern Society sends me spores of 4 Lycopodiaceae species. I attempted to get spores to germinate on 3 different formulations. One was a Knudson medium, one was called Parker Thompson Fern medium, and one was called Murashige fern medium. I poured plates of each medium. I took the tiny spores of each species and subjected them to a vacuum treatment to force water into them, a 10-minute bleach bath followed by a rinse with sterile water, and being plated. The vacuum treatment is a really cool process btw. It works by using water with a surfactant, which is a fancy term for something that resists floating by objects. Add a bit of detergent to some water and add spores. Then, swirl the solution and subject to vacuum pressure. Air is evacuated from the vessel and water enters its void, including the microscopic voids in the spore surface. This causes the spore to take up water, swell, and sink. This is CRUCIAL for surface-sterilizing the spore. Remember that tissue culture must be done in a sterile environment, and unbleached fern spores are anything but sterile.
So I then took my imbibed spores and put them in a solution of 10% bleach and a little surfactant to make sure the bleach really got onto that spore. I let the spores soak for 10 full minutes. I filled the bleaching vial to the brim, and then I surface sterilized the capped vial and put it within the flow hood. I shook it back and forth a few times to resuspend the spores. I have discovered you can make your own sterile rinse station for the next step. Get a pint mason jar. Fold a coffee filter in fourths and stuff it in the jar and cap it. Put the jar ring on loosely and autoclave/pressure cook for about 1 hour. Tighten the jar ring and then, when you must use it, you just pull out the coffee filter, make it into a cup, fit it over the rim and that sterilized ring will hold it in place! The air inside of the jar is also sterile (and yes it matters). I poured the bleach with spores into the filter after the appropriate time and rinsed them with sterile water. Then with a sterile pipette I dripped some spore water over the media and swirled the plates before sealing with parafilm, labeling, and storing in the dark.
Let’s pause with this story line and introduce another one. Whittier and his work, to my knowledge, didn’t ever see sporophytes forming in the species I’m currently working on. I wonder if using the medium I had success on (which I will reveal soon) but with different amounts of glucose or more nitrogen or something would help. I, at present, have yet to see sporophytes. I have more spores, and I plan on doing experiments with different concentrations of different things (the glucose concentration of the medium I experimented with earlier was 0.5%, or 5 grams of glucose per liter of medium) such as glucose and nitrogen to see if that influences germination rate, time, or sporophyte formation. I’ve read that similar species’ spores germinate better at a low glucose concentration but the subsequent gametophytes grow more rapidly at a higher concentration. Also, glucose is better for these gametophytes than sucrose is. I do not, however, know what is required for the formation of a sporophyte.
It took about a year. I kept the petri plates without any germination for an additional year. But 11 months after I sowed the spores, I noticed a few tiny whitish blobs on 1 medium. They were maybe the size of a grain of salt. I reckon each plate had 10,000 spores and only a handful of gametophytes showed up. Which basically means there was a 0.01% germination rate. Ok maybe more like 0.05%. The only medium with any germination after 2 years was the Parker-Thompson fern medium. A few months later, I put some of the cultures in the light and kept some in the dark. This was because I suspected that light would trigger the gametogenesis necessary for sporophyte formation. So this section will be dedicated to those 4 species. What did I observe with them? Did any make it to sporophyte? Hehe let’s see!
Dendrolycopodium dendroideum
I just wanna say that I have this species near my childhood home and I’d love to take some spores of it one day. I did get spores from Brian Aikins so I will have things to report on from this species.
Lycopodium clavatum
I almost got shot over this species. Why? Well because I was under the impression that we NEEDED a member of this genus for a teaching lab at URI. I had sections of D. dendroideum, which had formerly been classified as a Lycopodium. Not knowing this, I wandered into a shooting range where I found a clump of actual Lycopodium. I have gametophytes of this species at present.
Diphasiastrum digitatum
Diphasiastrum tristachyum
