The Perils of Plant Monogamy

Biology concepts – pollination, single pollinator, co-evolution, co-divergence

What’s bugging her, it’s supposed to be a party!

Imagine the best party of the year – it’s cold outside, but hot inside. The food is great, all your friends are there, everyone is receptive to flirtations by the opposite sex, it lasts for two nights in a row where it is, and then picks up in a new location again and again. Where is it and how do I get invited?

Last week we learned that the Philodendron selloum flower becomes endothermic for a 2 day period each year in order to facilitate its pollination. In this state, it attracts a single species of beetle, which parties down inside the flower and then takes the pollen to the next party - sort of BYOP.

The heating of the P. selloum spadix evaporates and spreads a pheromone that attracts male Cyclocephala beetles. Pollination by beetles (cantharophily) is not one of the most common mechanisms for the spread of pollen to ovules; pollination by bees (melittophily), butterflies (psychophily), or even the wind (anemophily) is more common. In most cases of cantharophily, the flowers are big, white, strong-smelling, and the male flowers are usually eaten but the ovaries are protected. This is exactly the scenario in P. selloum.

When the male beetles enter the flower, the angle of the spadix makes the male flowers available to be eaten, while the covering of the spathe discourages the beetles from leaving the pit. The nectar, pheromone, and flowers help draw the beetles in, but it is really the atmosphere and the company that keep them there.

Female beetles are drawn to the warm temperatures as well, as this affords the beetles the ability to feed and mate through the night, when the ambient temperature outside the flower would require them to slow down their activity (being ectotherms).

The Cyclocephala beetles, but there are other examples.

Orchids are famous for finicky polli beetles follow the

pheromone back to the flower, and after partying, the

flower closes down and kicks them out.

As the party winds down in this first P. selloum, the temperature is reduced and the spathe starts to close down on the spadix, forcing the beetles out – it’s closing time, you don’t have to go home, but you can’t stay here (lyrics by Third Eye Blind). As the beetles leave the flower two things happen, first they pass the viscidium, which coats the beetles with a sticky substance, and then they pass by the pollinia, which covers them with pollen grains.

The next night, a new P. selloum is ready to open its doors for the party. When the previous night’s revelers show up with their coating of pollen, they head directly to the nectar bar at the bottom of the pit, right where the female flowers are located. While getting their first drink of the night, they deposit the pollen on female flowers that reside there and pollinate the plant. The beetles do the work, but their rewards (increased mating time, increased food) are as important to them as the pollen is to P. selloum.

Pollinators can various animals.Non-animal 

pollinators work in some cases too,

such as wind and water.

This is one type of pollination party, but not the only one. Most plants invite a variety of different pollinators, or a plant might self-pollinate. In some orchids of colder regions where pollinators are particularly rare, self-pollination can be a last resort. If the flower is not visited by any pollinator, the caudicle (the stalk on which the pollen resides) will shrivel up in a particular shape, dropping the pollen directly on the stigma (containing the eggs).

Most plants invite pollinators of different species.  One bee may be a particularly effective pollinator of a particular plant, but that plant is probably also visited by a fly, a butterfly, a bird, a beetle, etc.  Few plants have a single pollinator, but P. selloum is one exception. While Cyclocephala beetle may pollinate other plants as well, it is the only species that pollinates P. selloum.

Single pollinators provide advantages to the plant. The need to attract only one species reduces the energy a plant must expend to attract multiple pollinators. Some pollinators are attracted to color, some to different scents, some to different UV patterns, some to different nectars. To draw different pollinators the plant will have to have many attractants, and this costs energy. 

The more pollinators a flower depends on, the more energy the plant must spend on attractants. For instance, some flowers use color and UV patterns, as on the left. Some flowers use nectar and visible light patterns, as with the pitcher plant. The red flower is rafflesia, the largest flower in the world. It smells like rotting flesh to attract flies. Some flowers use mimicry, like the bee orchid on the right, it looks like a female bee and attracts males that will try to mate with it.

Another advantage is seen after the pollen is gathered on the pollinator. In order for a pollination to be successful, the pollen must be delivered to the female organs of a plant of the same species. If a pollinator species has developed a relationship with a certain plant (attracted by a specific odor or color, etc.) then the chances are higher that it will visit another plant of the same species after gathering pollen. This increases the chances of cross-pollination.

The dependence of a plant on a specific pollinator amplifies the plant’s vulnerability if there is a decrease in the pollinator numbers. It has no second option for pollination. This is one reason why cross-pollination is preferred to self-pollination. Evolution does not anticipate the future; it proceeds as if the pollinators are present in good numbers. The plant needs the greatest diversity of gene mutations and rearrangements in order to adapt to unanticipated changes in pollinator number, behavior, or preference. This diversity is provided by cross-pollination with another plant, not by reiteration of existing gene patterns by pollination with the plant’s own genome.

The disadvantage of employing a single pollinator is becoming more obvious. Recent years have seen large decreases in wild pollinator populations. Honeybees have experienced colony collapse disorder, and 2011 figures indicate that 10% of American bumblebee species are near extinction. More than 40 species of pollinating insects in the US are endangered, and even more shocking, 1200 vertebrate species of pollinators are termed “at risk.” If this many pollinator species are in decline, even plants pollinated by multiple species might feel the pinch. Can you imagine how many plants that rely on a single species of pollinator might be in danger of extinction?

Many orchids use a single pollinator. Orchids are the most 

diverse flower group, as show by the Dracula orchid, 

the spectacle orchid, and the small tongue orchid, left to right.

P. selloum is an interesting exception to the rule of multiple pollinators, but there are other examples. For instance, orchids are famous for finicky pollination. There are >25,000 species of orchids, the largest group of plants (in contrast, all birds represent only ~10,000 species), and single pollinators are responsible for the propagation of thousands of them. In South America and South Africa, the number of single pollinator species is quite high, including many orchid species. (Why? I have no idea.)

The specific interactions between the single pollinator and the plant it pollinates are often a result of co-evolution. In technical terms, this means describes the reciprocal natural selection and evolutionary change that occurs between two species by exertion of selective pressure on each other. The two species could be trying to outfox one another, like a parasite and its host, or could be working together, like the pollinator and pollinated.

As the two interacting species interact, they may evolve so that they rely solely on each other for that particular interaction. This could also lead to each species diverging from its closest relatives. This particular type of co-evolution is called co-divergence.

Co-divergent speciation can be seen in the host parasite relationship between the malaria parasite, Plasmodium falciparum, and humans. When humans and chimps diverged (about 4-7 million years ago), some P. falciparum evolved to infect only chimps, while others followed human evolution and became specific for humans.

The Darwin hawk moth wasn’t known when the star orchid

was first described. Charles Darwin just predicted it must exist.

Predicting the existence of a moth with 35 cm tongue didn’t win   
Darwin many fans, but he was right.

In similar fashion, there are numerous plants that have co-evolved with a pollinator. The Angraecum sesquipedale orchid (star orchid) is a classic example. Charles Darwin was sent several examples of this flower and described them in an 1862 publication. Darwin noted that the nectar of this flower was located deep within a hollow spur. To reach the nectar, a pollinator would bump into the pollen and it would stick.

 

However, the tube was so narrow, that no known insect could have been considered a pollinator of this plant. Darwin predicted that an insect with a 30-35 cm proboscis (tongue-like appendage) would be found pollinating A. sesquipedale. He was ridiculed for such a bold proposition, but 40 years later, just such an insect was discovered, the Xanthopan morganii praedicta moth (named for Darwin’s prediction).

Nature is full of exceptions to the rule of multiple pollinators, including snapdragons that need a bee of specific weight to trip the opening mechanism of the flower. Several orchids that use the same single pollinator place the pollen on different parts of the pollinator’s body, so that female flowers of the same species will come into contact with the correct pollen. These are still exceptions, as the vast majority of plants use multiple pollinators – they just aren’t as interesting.

We have seen a plant that can become endothermic in order to pollinate. Next time we will look at a mammal that has gone the other direction, but for the same reason - survival.

Chupp AD, Battaglia LL, Schauber EM, & Sipes SD (2015). Orchid-pollinator interactions and potential vulnerability to biological invasion. AoB PLANTS, 7 PMID: 26286221 Whitehead MR, & Peakall R (2014). 

Pollinator specificity drives strong prepollination reproductive isolation in sympatric sexually deceptive orchids. Evolution; international journal of organic evolution, 68 (6), 1561-75 PMID: 24527666

For more information, classroom activities and laboratories on P. selloum, pollinators and co-evolution, see:

P. selloum –

http://www.floridata.com/ref/p/phil_bip.cfm

http://faculty.ucc.edu/biology-ombrello/pow/Philodendron.htm

http://www.exoticrainforest.com/Philodendron%20bipinnatifidum%20pc.html

http://www.pburch.net/plants/C3397731/E20060506081651/

pollinators –

http://www.pbs.org/wgbh/nova/nature/pollination-game.html

http://pollinatorlive.pwnet.org/teacher/lessons.php

http://www.smithsonianeducation.org/educators/lesson_plans/partners_in_pollination/lesson1_main.html

http://www.nebraskawildlife.org/ww08-PollinatorCurriculum.htm

www.mbgnet.net/bioplants/downloads/pollination.pdf

www.nybg.org/files/pollination.pdf

www.ableweb.org/volumes/vol-22/12-puterbaugh.pdf

www.ppdl.purdue.edu/ppdl/expert/Pollinate_Tomato_Pumpkin.html

www.infocusmagazine.org/6.3/env_pollinators.htm

www.pollinator.org/Resources/facts.Gardeners.pdf

co-evolution –

www.pbs.org/americanfieldguide/teachers/insects/insects.pdf

http://www.teachersdomain.org/resource/tdc02.sci.life.evo.lp_speciation/

www.stanford.edu/group/stanfordbirds/text/essays/Coevolution.html

http://www.ucl.ac.uk/~ucbhdjm/courses/b242/Coevol/Coevol.html

http://evolution.berkeley.edu/evosite/evo101/IIIFCoevolution.shtml

http://bioweb.cs.earlham.edu/9-12/evolution/HTML/converge.html

http://www.universityofcalifornia.edu/news/article/4463

http://www.teachersdomain.org/resource/tdc02.sci.life.evo.lp_speciation/

I’ll Fly Home—Or Not

The snowy owl is sedentary, meaning it does not

migrate. The males are almost perfectly white,

while the females and juveniles can have black

barring. They sit, look, and listen for their prey,

which includes small rodents and even other birds.

Their hearing is good enough to let them target a

mouse under the snow from hundreds of yards away.

The Arctic tern travels from north of the Arctic Circle to Antarctica and back again every year. On the other hand, the Snowy Owl lives in the arctic region year-round; it doesn't migrate at all.

The Question of the Day:

Why do some birds migrate while other birds stay in one place?

The possible explanations are many. Maybe the type of food they eat is present only part of the year, or maybe they can’t stand the cold temperature. They might need to have their babies in a place away from predators, or perhaps migration is an evolutionary holdover that had a reason in the past, but no longer is necessary.

How could you start to determine the reason for migration in only some bird species? I would start by looking at how closely related the migratory and non-migratory species are. Maybe all the birds that migrate are more closely related to one another than to the birds that don’t move around during the year. This would suggest that there is a genetic basis for why only some species migrate.

One research group did just this in 2007. The looked at 379 species of flycatchers, a closely related group of birds. They found that almost equal numbers of species were migratory or resident, so it doesn’t appear that genetic relatedness is the answer.

The painted bunting is among the most colorful

birds in North America. This male has several colors,

while the female is a brilliant green. They breed in

south central US or on the southern east coast, but

winter in Central America; therefore, they are

seasonally migratory.

Maybe the need to migrate has to do with the geographic region. About 90% of birds in the arctic migrate, some are present there only in the mid-summer months. The arctic tern is a good example. Arctic terns move with the summer, breeding in the arctic in May-July, moving down along continental coasts to arrive in Antarctica for the months of December to February. The entire distance traveled could be as much as 32,000 km (20,000 miles) in a single year.

Similar to the arctic region, the east coast of North America has species that migrate and species that are sedentary. About 80% of the birds species of the east coast move south during the colder months, but on the Pacific coast, almost all the bird species are non-migratory.

So migration is not due to the type of geography around the birds. However, the east coast of North America does have larger temperature swings than the west coast, so maybe it is just that some birds can’t deal with the cold. 

Some non-migratory birds can control the amount

of blood that travels to the legs in order to conserve

body heat. This works even better if they can reduce

the amount of contact with the cold surfaces, so some

of these birds perch on one leg at a time.

Many birds that do not migrate have special adaptations to deal with the cold. Trying to keep a constant body temperature (endothermy) takes a lot of energy, and birds live right on the edge of having enough energy anyway. Flying requires a huge amount of energy and they must eat almost constantly just to keep enough carbohydrates in their system to be able to move around to find more food.

Burning more energy to keep warm might tip them over the edge into starvation. To alleviate this problem, many birds can allow parts of their bodies cool down to freezing or near freezing, while keeping their internal organs at a temperature that will preserve their function. Blood flow is a major way to keep parts of the body warm, a duck standing on the ice can reduce the blood flow to its feet and reduce the amount of heat lost to the cold ice. The duck’s chest may be 40˚C, but its feet could be just one degree above freezing.

But let us look again at the arctic tern. It migrates from the north polar region to the Antarctic region in such a way that it sees two summers each year. But these are summers in name only. The arctic summer has an average temperature from -10˚C to 10˚C, so much of the time the tern is there, the temperatures are near zero.

The arctic tern has a ghastly commute each year.

The trip is even more amazing when you consider

that during its yearly molting, the tern flies very

little.  So, all that distance must be fit in to just a part

of the year, not the entire 365 days. I guess they

vacation by NOT traveling.

Then when they reach the Antarctic, the summer there has an average temperature of -2˚C to 2˚C. This is hardly a balmy vacation destination for the tern. The temperatures in both its breeding grounds and wintering grounds would require it to have elaborate temperature control and energy-saving adaptations. Therefore, inability to tolerate cold temperatures is not the reason for migration, at least not for many birds.

The group who carried out the 2007 study concluded that the main reason that only some flycatcher species migrate is not due to what they eat, or when they breed, or what is trying to et them, but to how available their food source is. Whether they are fruit eaters, or insect eaters, or seed eaters, how easy it is to find their food is the most common reason that migration has evolved for a specific species in a specific location.

The food availability hypothesis is supported by certain types of migration that are common in North America. Irruptive migration is characterized by a population moving to another place, but there is no yearly, seasonal, or geographic pattern. The birds may migrate one year, then not again for a dozen years, or they might go for several years in a row. North American seed-eating birds are famous for these migrations. The distance and number of individuals that migrate are also not very predictable, and this all makes it sound like the movement is linked to food availability. However, it could also be to escape some population explosion in a predator species or for some other reason.

It isn’t only birds that might undergo partial migration.

Some crab species will migrate for breeding purposes,

like these Christmas Island Red Crabs.  Individuals

that won't breed just don’t make the trip. They may be

too young, too old, too lazy. In other cases, when

populations migrate away from the breeding grounds,

some individuals may remain there the year round.

Another type of migration is partial migration, a pattern wherein not all birds of a species in a certain location will migrate, only some of them leave in non-breeding times, while others stick around year-round. Food may be available for some, but not all, or the environment may be unsuitable for some weaker individuals to have enough time to forage for a sufficient amount of food. These (and other reasons) might explain why partial migration exists, but one question remains, who stays and who goes? The choices could be based on age, altruism, suitability, dominance… laziness?

As an aside, it isn’t just birds that migrate. Mammals move from place to place, sometimes with a defined pattern during the year, but sometimes they just follow the food, a process called random migration. And some insects migrate as well.

For many years, the migration of the Monarch butterfly was believed to be the longest insect migration. But this is not the typical migration we think of, where an individual moves from one place to another and then back again. The migration of the monarch butterfly takes four generations to complete. Some generations are born and fly a long distance to lay their eggs, while others are born, live, and reproduce in a small area. But altogether, this butterfly moves from as far north as Canada to the high mountains of Mexico and back each year, about 7000 km (4400 miles).

Globe skimmer dragonflies breed in freshwater pools,

so they migrate from India’s monsoon season to the rainy

season in East Africa, all in search of a place to lay

eggs. They make stopovers on the Indian Ocean islands,

but only to rest, because there are very few

pools of freshwater on these coral cay islands.

A few years ago, a biologist in the Maldive Islands started to wonder about the movement of globe skimmer dragonflies where he lived. They seemed to be plentiful in some periods and absent in others. He started to track them, and found that they have an even larger migration pattern than the monarch butterfly. What is more, they fly long distances over the ocean with no place to stop and rest.

Over a series of generations, the dragonflies move from India to the Maldives, some 600-800 km across the open sea. Then they move to east Africa, from Uganda to Kenya and Mozambique. In January, they start back toward India, and complete their migration of more than 18,000 km (>11,000 miles).

So birds may get most of the publicity, but insects hold their own in the migration game. Of course, it does take four generations of dragonflies or butterflies to make their complete journey, where a single arctic tern may make its entire 20,000 mile trip thirty times in its lifetime. O.K., they are both pretty impressive when you consider most people need a car to go down the street to the grocery store and back.



Davenport LC, Goodenough KS, & Haugaasen T (2016). Birds of Two Oceans? Trans-Andean and Divergent Migration of Black Skimmers (Rynchops niger cinerascens) from the Peruvian Amazon. PloS one, 11 (1) PMID: 26760301

Ahola MP, Laaksonen T, Eeva T, & Lehikoinen E (2007). Climate change can alter competitive relationships between resident and migratory birds. The Journal of animal ecology, 76 (6), 1045-52 PMID: 17922701

Hobson KA, Anderson RC, Soto DX, & Wassenaar LI (2012). Isotopic evidence that dragonflies (Pantala flavescens) migrating through the Maldives come from the northern Indian subcontinent. PloS one, 7 (12) PMID: 23285106

Big Bugs, Little Bugs

The titan beetle (Titanus giganteus), is not necessarily

a gentle giant. Its jaws can snap pencils and easily cut

into human flesh…. and they fly. Calm down, the adults

don’t feed, they just look for mates, so you won’t wake

to one nibbling your toes away.

Today, the biggest insects are goliath beetles, atlas moths, and giant stick insects. But during the carboniferous period (360 -300 million years ago), there were millipedes that were 2 m (6 ft) long and dragonflies (order Protodonata) the size of eagles!

Today’s question is:

How did insects get so big in the carboniferous period, and if they were big once, why are they so much smaller now?

It wasn’t just the insects that grew large way back then, the first large plants flourished in this same time period. Some ferns grew to be 20 m (65 ft) or more in height, and the diameters of trunks were increased. While not as big as today’s largest plants, the change was significant, as plants before this period did not exceed 3 to 5 ft in height.There was plentiful carbon dioxide in the atmosphere and the environment was warm all year round. This allowed lots of photosynthesis and lots of growth.

Nice picture to give scale, but humans and the

Arthropleura never co-existed. Even though the

species alive today aren’t as big, you still have

to beware. Many centipedes are venomous and

some millipedes can emit hydrogen cyanide gas.

Plants were evolving lignan in this carboniferous period (carbonis = coal, and ferrous = producing). Lignan is the stiffest of the plant molecules and is what allows them to grow tall.  This is also what gives the carboniferous period its name, as the lignan of plants is the major component of the coal that formed from their remains.

Horsetails, another type of plant of the carboniferous age and which are still around today, also grew much bigger. Horsetails today do not get any taller than about 1 m (3 ft), but in the carboniferous period, they were often 10-15 m (33-48 ft) tall.

But it was the arthropods that get all the publicity. Scorpions almost a meter in length deserve to have a lot of attention paid to them! And consider the yuck factor of a 7 inch cockroach scurrying around at your feet.

The plants got big during the Carboniferous period.

Some lycophyte trees were 30 m (98 ft) tall and had

trunks of 2 m (6.5 ft) diameter. Their closest

relatives today are the club mosses, which are about

20 cm or less in height. Talk about deflating your ego.

What allowed these animals to grow so large? Scientists think it was related to the lignan. With lignan, the plants could grow larger and support more photosynthetic material. The carboniferous period is when the first forests appeared.

With more and bigger plants, more carbon dioxide was converted to carbohydrate, and more oxygen was produced as a result. The oxygen content of the air in the carboniferous period reached levels of 35% or more (today it is about 21%).

More oxygen in the air meant that more oxygen could be transferred into the blood of animals.  They could carry out more oxidative phosphorylation and produce more cellular energy (ATP), especially since there was all this plant material around to eat to gain carbohydrates (or plant-eaters to hunt down and eat). This growth spurt especially applied to animals without traditional circulatory systems. Insects, for instance.

Insects use spiracles on the sides of their bodies to take in air. The oxygen and other gases are moved through a system of smaller and smaller tubes (called trachea) to bring the oxygen to all the cells of the body. The carbon dioxide produced during cellular respiration is removed in the same way.

The spiracles of a flea are on the side of its abdomen

and the air travels through the tracheae to bring

oxygen to every cell. It seems like he would drown if

he took a dip in the hot tub.

This is not a particularly efficient way to move gases in and out of cells. A slightly bigger bug must have a much more voluminous system of trachea, and at some point, the respiratory system would have to be bigger than the entire volume of the insect! There would be no room for all the other organs. But with a high concentration of oxygen, the spiracle/tracheae system is efficient enough, and the insects can grow very large.

High oxygen in the air also meant high oxygen in the water. Carboniferous era fish and amphibians grew large as well. Some toothed fishes of this time were impressive predators, and were more than 7 meters (23 ft) in length. Isopods in the oceans were also huge. Even today some of these crustaceans can be pretty impressive. Bathynomus giganteus can grow to more than over 16 inches in length. 

So big plants brought big oxygen levels, which brought big animals. But why are the arthropods so much smaller today as compared to then? Well, the oxygen levels are lower now, so according to a 2006 study the inefficient spiracles system could not support the large body. Insects had to get smaller.

Here is an isopod that grabbed a ride when a deep sea remotely operated 

vehicle was recovered. They look even creepier from the front with 

silver eyes. Isopods are related to shrimp and crabs; I think we’re going 

to need much more butter and lemon.

But there is an additional hypothesis that may also contribute to the small size of many insects, especially flying insects. According to a 2012 study, the size of flying insects is related to another aspect of oxygen. When the explosion of plants in the carboniferous period raised the oxygen levels, the air became more dense (oxygen is a heavier gas). The insects were able to become better fliers, since their wings could move more air.

The data says that one reason flying insect

got smaller was to avoid being easy catches for the birds

that were becoming better fliers and hunters. Really?

Name me a couple birds that would go after this guy if

he was flying today.

Millions of years later, birds evolved. As they became better fliers (their ability was also based on their ability to move air over their wings), they became better hunters. Better hunting birds could catch flying insects (and terrestrial insects for that matter) better. So it became a disadvantage to be big. The smaller insects now had a reproductive advantage; they were the only ones surviving to have offspring. Over a period of time, the insects grew smaller on the whole.

So today we have fairly small arthropods and insects, although my wife has personally never seen a small insect. According to her they are all large enough to carry off small children and have evil looks in their eyes.

Matthew E. Clapham1 and Jered A. Karr (2012). Environmental and biotic controls on the evolutionary history of insect body size Proc. Natl. Acad. Sci. USA DOI: 10.1073/pnas.1204026109

Next week – ideas for long studies on the nature of science.

It’s All in the Numbers - Sizes in Nature

If all the animal species are broken up into groups, the light 

blue section includes insects, and the rest of the 

circle colors represent every other animal on Earth!

Comparisons help to make very big or very small numbers meaningful, and biology is chock full of big and small numbers. For instance, there are more insects in the world than there are humans. By more, I mean ALOT MORE, something like 1.5 x 10¹⁸ insects. But what does that number mean? Consider looking at it this way; the world population hit 7 billion last year and that's a big number, but even if we were to double our population again in the next ten minutes, there would still be 100 million insects for every human on earth. This certainly makes an impression, but it seems small when comparing the most numerous organisms, bacteria, to humans.

Bacteria outnumber us by orders of magnitude more than insects do; they live everywhere, in every environment. They have been found in 0.5 million year-old permafrost as well as 40 miles up in the atmosphere. There are approximately 100 million to 1 billion bacteria in every teaspoon of dirt, so in total there are currently 5 x10³⁰ bacteria carrying out their daily routines. That means there are about 5 x 10¹⁹ living bacteria (that is 50,000,000,000,000,000,000) for every person who has EVER LIVED. Another way of visualizing this might be to imagine that each bacterium is a penny being stacked. The column would be a trillion light years high. That’s about five times the diameter of the observable universe.

Nanobacteria are still controversial, the 0.2 µm diameter is 

close to the smallest size that could still hold DNA. 

For comparison, the white line in panel A is 1 µm long, 

and in Panel C the line is just 0.1 µm.

While the redwoods might be slightly taller than the sequoias, 

the mass of the sequoias is much greater because the 

trunks have such a large diameter.

Even using comparisons and analogies, these numbers are almost too big to comprehend. It isn’t much easier when talking about sizes. The scale of life is amazing, from the smallest bacteria (called nanobacteria), just 0.2 µm in size (1/5,000,000 of a meter), to the biggest living thing on Earth, a Giant Sequoia called General Sherman. This behemoth of a tree is more than 83 meters (272 ft.) in height and 1,225,000 kilograms (2,701,000 lb.) in mass. This means that from smallest to largest, life spans more than eight orders of magnitude. In terms of biomass, the difference between the smallest bacterium and General Sherman is even greater, about 1 x 10²³, about the same as difference in mass as one human compared to seven Earths.

On a smaller scale, the difference in size between bacteria and nucleated cells (eukaryotic cells) is still pretty stunning. A single macrophage cell of your immune system can ingest more than 100 bacteria without flinching, and macrophages are nowhere near the biggest eukaryotic cells. These different sizes demand some distinctions in how cells conduct their business; for example, how they move molecules into and within themselves.

A macrophage reaching out and ingesting bacteria.

The bacteria are the small, connected rods.

Eukaryotic cells, unlike prokaryotic cells (bacteria and Archaea), have specialized systems, like actin filaments, cytoskeleton, and microtubules. These apparatus are designed to act like conveyor belts; they carry different molecules through the cell to their needed destinations. Eukaryotes also have specific receptors for bringing in specific molecules. These are fast systems of uptake and movement, and can work against a concentration gradient.

The cytoskeleton of the eukaryotic cell stretch out like fibers.

They help it move, can convey molecules from place to place,

and holds the cells shape.

Unfortunately, bacteria only have diffusion to move molecules around their insides. This makes things doubly hard on them because bacteria have limited access to resources; most often they meet up with few molecules that are important to them (being a small cell in a big environment). Therefore, they need to get as many of these resources into their cell as possible and move throughout their entire volume quickly.

Diffusion is the movement of molecules from places where there a lot of them toward places here there are fewer of them (from high concentration to low concentration). Think of a crowd pouring out onto the football field after a big win. You start with many people in the stands and very few on the field, but end up with about an equal number of people in all parts of the stadium. Bacteria count on consuming their nutrients this way. Important molecules diffuse into the cell, and then get metabolized for energy or other building blocks. This breaking down and reassembly of molecules helps ensure that the concentration of important molecules is always lower inside the cell, so diffusion into the cell can continue. Importantly, as the width or length of a cell doubles, the volume increases by a factor of eight; therefore, prokaryotic cells remain small so that they can get molecules everywhere they need them quickly. It is the only way for diffusion to remain profitable for them.

Diffusion is the movement of from where there are 

many to where there are few. If it is water 

molecules that are moving, then call it osmosis.

Diffusion is not quite as simple as people pouring out the stands. There are several aspects of this process that are important to bacteria. The first of these is the diffusion rate, which is based on a diffusion coefficient for each different molecule, and the liquid it is moving through. For oxygen moving through water, the diffusion rate is about 1 mm/hr. This means that for an average sized bacteria it only takes 1 millisecond (1/1000th of a second) for an oxygen molecule to travel across the entire cell.

There is also the mixing rate; this refers to the time it takes for a molecule that enters the cell to have an equal probability of being found in any part of the cell. A 1µm (1/1,000,000 of a meter) bacterium has a mixing time of roughly 1 millisecond. But since the volume increases by a factor of eight as the size doubles, it would not take much growth for the mixing time to become problematic if a cell was to rely on diffusion alone.

Finally, there is the issue of traffic time. Every reaction that takes place in a cell involves two or more molecules finding one another and then interacting. In both prokaryotic and eukaryotic cells there are some systems designed to help bring molecules together, but in the end, it is basically luck – they have to run into one another. The number of molecules can affect this time; say you want molecule A to meet molecule B. If the cell contained only one of each molecule, this could take a while, but if there are 1000A’s and 1000B’s, then the traffic time will be decreased considerably. For average sized bacteria, traffic times exist in the range of 1 second, but again, if they are much bigger, the chances of molecules meeting their partners goes down dramatically.

If the bacterium grows too big, the diffusion rate, mixing time, and traffic time can become too long to permit survival. Therefore, size limitations seem to be set for bacteria. However, some bacteria just have to be rule breakers. There are two excellent examples of bacteria that have evolved ways to overcome the diffusion problems associated with increased size, and we'll start to look at them next week.



Schulz, H., & Jørgensen, B. (2001). Big Bacteria Annual Review of Microbiology, 55 (1), 105-137 DOI: 10.1146/annurev.micro.55.1.105



For more information on numbers in nature, diffusion, and cytoskeleton, as well as web-based activities and experiments, go to:

Cell size and volume:
http://staff.jccc.net/pdecell/cells/cellsize.html
faculty.massasoit.mass.edu/whanna/121_assets/15-week_2_prelab.pdf
http://www.youtube.com/watch?v=qdvKM1m0jnE
http://www.cellsalive.com/howbig.htm
www.nsa.gov/academia/_files/collected_learning/high.../surface_area.pdf
www.smccd.net/accounts/bucher/modules/DuzSizeMatter.pdf
http://www.accessexcellence.org/AE/AEC/AEF/1996/deaver_cell.php


scaling in nature:
http://www.nature.com/scitable/content/the-sizes-of-organisms-span-21-orders-15321100
http://learn.genetics.utah.edu/content/begin/cells/scale/
http://www.dnatube.com/video/596/Size-Analogies-of-Bacteria-and-Viruses
http://www.smithsonianeducation.org/educators/lesson_plans/size_shapes_animals/index.html


diffusion:
http://www.biologycorner.com/bio1/diffusion.html
http://highered.mcgraw-hill.com/sites/0072495855/student_view0/chapter2/animation__how_diffusion_works.html
http://staff.jccc.net/pdecell/cells/diffusion.html
http://hyperphysics.phy-astr.gsu.edu/hbase/kinetic/diffus.html
http://www.wisc-online.com/objects/ViewObject.aspx?ID=ap1903
http://www.biologycorner.com/2009/09/16/diffusion-lab/
http://chem.lapeer.org/Bio1Docs/Diffusion.html
http://www.biologyjunction.com/osmosis__diffusion_in_egg_lab.htm
http://phet.colorado.edu/en/contributions/view/3415


cytoskeleton:
http://www.cellsalive.com/cells/cytoskel.htm
http://www.youtube.com/watch?v=5rqbmLiSkpk
http://www.biochemweb.org/cytoskeleton.shtml
http://www.biology.arizona.edu/cell_bio/tutorials/cytoskeleton/page1.html
http://www.biology.arizona.edu/cell_bio/tutorials/cytoskeleton/main.html
http://www.youtube.com/watch?v=zlYyoi5vpE8

What’s So Repelling About Repellents?

Biology concepts – thermosensing, repellent, odor receptors, gustatory receptors, semiochemcials

Science explains our world, and then technology and engineering

build a model of that for our use. The better we know how our

universe works, the better we can make use of it. In the 1985

film Real Genius, this difference is stated when the scientist

students ask what a 6 megawatt laser might be for, one student

says, “Let the engineers figure out a use for it.” In this case, they

used it to fill a house with popcorn.

Science exists to describe our universe in terms of rules and mechanisms; what is and how it comes to be. Knowing that something exists is only half the equation. Science seeks to explain how something exists in terms of the rules of the universe. Observation is good, but it only shows us the question – mechanisms of action and interactions show us the answers.

As an example – we know that certain naturally occurring oils and well as some man made chemicals keep mosquitoes from feeding on us. This is the observation. But the question is – how do mosquito repellents work? The answer is more interesting and more complicated than you would initially think. Repellents rarely repel.

Investigating how chemicals keep us from getting bitten will teach us about how the living systems work, will give us a better understanding of our universe, and then give us better insect repellents. Don’t think that’s important? Consider the hundreds of millions of people who are infected every year (several million die) with mosquito-borne diseases (malaria, encephalitis, dengue fever, yellow fever, filiariasis). So yes, we need more repellents.

Mosquito borne diseases can be unpleasant at best. Top left is

filariasis, a worm is transmitted via mosquito and it clogs up

your lymphatic vessels, so that body parts swell from excess

fluid. Top right – malaria can result in so much red blood cell

lysis that your spleen (the guy who cleans them up) can

rupture. Bottom left – Dengue fever is often called breakbone

fever, the pain is not something an image can express. But the

hemorrhagic form of the disease can produce some bleeding

in weird places. Oh, and it can kill you too. Bottom right –

yellow fever is caused by a virus transmitted by mosquito. Your

liver breaks down and causes your whole body to turn yellow

and you bleed into your skin.

We should start with the repellents for which we have good ideas of their mechanism of action. But there aren’t any. We have some hypotheses and working ideas of the modes of action of mosquito repellents, but nothing is definitive yet. Let’s look at two of them and see if we can find some common pathways.

Citronella oil

Citronella is a combination of many different natural oils produced in lemongrass plants (Cymbopogon nardus and Cymbopogon winteratu). As a natural oil and a flavoring in Asian cooking, one would think that citronella oil would be considered just about the safest insect repellent this side of a slap with an open palm.

But no, Canada says that one small component of citronella oil called methyleugenol, can increase the likelihood of tumor formation in rats. Of course this was when methyleugenol was distilled from the oil, given by itself in large doses, and introduced directly into the stomach. But Canada is still in the process of banning citronella oil as an insect repellent. Of course, you can still eat thai food in Canada, which is often flavored with lemongrass.

The EU, on the other hand, said that the repelling function of citronella oil hadn’t been proven and it was deemed illegal to use in the EU in 2006. Oh, you could eat it, and use it soap or perfumes, you just couldn’t use it to keep mosquitoes away. They reconsidered in 2014 and some restricted uses of citronella oil as a repellent are now allowed.

Citronella oil comes from the lemongrass plant (Cymbopogon

nardus or Cymbopogon winteratu). There are two major species

for acquiring the oil, and the oil from each is a little different in

the percentage of each chemical. Lemon grass is also used in

cooking, the woody stalks are used with extra long cook times.

The torches that burn citronella oil work pretty well, but you

have to stay in the volatilized cloud of oil for them to be efficient.

Despite these issues, the U.S. Environmental Protection Agency (EPA) says citronella is safe and effective as an insect repellent. One weird side issue – you can take all the lemongrass you want from the US to Canada, where its oil is under attack, but you can’t bring any lemongrass from Canada to the US, where it is considered safe. Hmmmm.

Citronella oil probably works in a couple of ways. It's strong and sweet smelling, so it covers up and dilutes the odors that mosquitoes use to find you. If they’re detecting all the citronella in the air, then they aren’t smelling you. But research also shows that citronella oil activates TRPA1 ion channels. In us, they detect cold and noxious chemicals and are interpreted as pain. It is very possible that the detected signals in mosquitoes just come through as something unpleasant and to be avoided.

In this way, citronella would be an actual repellent. It repels on contact as well, as the taste is thought to activate bitter taste receptors and contact greatly reduces feeding time.

But citronella only seems to work when you are in the cloud produced by burning the candles or torches, or within the area of the spray. And if you’re using an oil or cream with citronella, it should really be reapplied every 30-45 minutes - not the most user-friendly method for discouraging pests.

DEET

World War II in the Pacific was an insect nightmare for the US Army. In response to the plethora of insect-borne disease that ran through the allied forces, defense scientists starting looking for better insect repellents. In 1946, their efforts produced N,N-Diethyl-meta-toluamide, or DEET.

Just how they came up with DEET is a mystery to me, it must have been a massive exercise in trial and error. Why? Because we know less about how DEET works than we do about citronella oil. And that’s with the benefit of 40 years of research. They didn’t have a clue how it worked or even what systems it was targeting when developed in the 40’s.

Guess which hand has been treated with DEET. The

mosquitoes come very close to the hand that was treated,

but don’t land on it. This argues that DEET is less repelling,

than it is disguising. On the right, the structure of DEET is

similar to several human semiochemicals, it fits into the lock

and key system of several odor receptors and activates or

inhibits them.

Originally it was believed that DEET disrupted the mosquito’s ability to detect semiochemicals (octenol) produced by mammals, especially humans, so mosquitoes couldn’t find a mammalian host to feed on. Then they played around with the idea that it blocked detection of CO₂.

More recent studies have been more rigorous, but haven’t helped solve the puzzle. A 2008 study suggested that DEET was actually repellent; the mosquitoes didn’t like the smell and would avoid it. But other studies have shown different mechanisms of action.

A study in the journal Nature in 2011 found that mosquito odor receptors could be confused by DEET. The receptors for octenol were less responsive in the presence of DEET, but other receptors more more responsive.  The conclusion of the study was that odorants from humans could be detected, but their pattern was confused, so the mosquito didn’t recognize the target as a target. It’s as if we disappear from the mosquitoes radar when we wear DEET.

A 2010 study showed similar results. DEET activated certain odor receptors but not others when given alone, but the opposite effects were seen when DEET was given in the presence of things from human sweat that would normally attract a mosquito. Once again, the signals were confused. This is really more of a chemical disguise for us, not a repellent. Next time your kids go outside, you should insist that they apply their mosquito confusant.

However, a 2013 study in the Journal of Vector Ecology found that heat and moisture were critical elements for recognition of targets by female mosquitoes, and that DEET messed not with odor, but with detection of heat and/or moisture. Different from the other studies, but still more of a masking than a repellent.

Something a little disturbing. Mosquitoes can learn to ignore

DEET. Most mosquitoes will be confused by DEET and never

find you. But if they do and then are repelled by the taste, they

learn from that and the second taste is not repellent. Hopefully

they just don’t find you a second time.

There was an interesting study from 2013 that showed that if you mutate or knock out Orco, one of the co-receptors (a protein that works with many different odor receptors so that they can function properly), then two things happened. One, DEET didn’t have any effect on the mosquitoes, and two, mosquitoes that normally preferred humans greatly would then settle for any mammal.

Weird - Orco is needed for both DEET to work and for mosquitoes to find humans more attractive. I haven’t figured that one out yet. The researchers showed that DEET only maintained an effect on the Orco mutant mosquitoes when they landed on a DEET covered surface, and then they didn’t like it at all.

This suggested that DEET might have more than one mechanism, confusion in the air and repellent taste on contact. Older studies supported this idea, as a couple of studies in 2005 and 2006 showed that contact with DEET would reduce feeding behavior in mosquitoes and one in 2010 showed that fruit fly bitter taste receptors are activated by DEET.

So, we have studies that say DEET is a confusant rather than a repellent, others that say it is a true obnoxious smell that they can’t stand, and yet others that say DEET is confusing to the smell and repellent to the taste. But there are more. Other studies suggest that DEET actually inhibits the smelling of anything, while others say that it inhibits an important protein called cytochrome p450.

Used commercially since the 1950’s, DEET has been the gold standard for efficiency for many years. Although it has to be used at fairly high concentrations, it can keep mosquitoes away for 4-6 hours at concentrations where citronella oil might work for less than an hour. At 100% concentration, DEET is active for more than 12 hours. What’s more, if you combine DEET with 5% vanillin, it works two hours longer!

A lime with cloves stuck in it as a mosquito repellent – really?

Well, lime is kind of like citronella oil, and clove has eugenol,

which acts on TRPV1 ion channels. But how many would you

have to have, or do you wear them like earrings? Penny royal

contains menthol and mosquitoes stay away from it. But it

also has toxins that will kill you.

As good as DEET is, people still question whether it’s safe. The EPA in a 2014 review said that DEET is safe for human use and poses no identifiable risks for human health, even in children. But this doesn’t keep people from suspecting chemical usage of carrying negative effects.

On the other hand, DEET dissolves plastic, foam rubber, spandex, gore-tex, and nylon. I can see where this might make people leery about slathering it on their skin for hours at a time. And a few people are allergic to DEET, so the best current repellent isn’t without some negatives.

One last point – a newer repellent called picaridin is almost as effective as DEET and doesn’t eat your back packing equipment and clothes. The interesting point is that picaridin is a synthetic version of piperine, the spicy chemical in black peppercorns. Add to this that menthol is also a fairly decent mosquito repellent, and we have some good arguments that TRP receptors might be involved in repelling activity – as with citronella oil. Piperine is a TRPV1 agonist, and menthol activates TRPM8 and TRPV1. All our talk about spicy food and heat/cold receptors has an impact even in the spread of malaria and other deadly diseases!

Next week, another question to answer - do sunflowers really turn with the sun?

DeGennaro M, McBride CS, Seeholzer L, Nakagawa T, Dennis EJ, Goldman C, Jasinskiene N, James AA, & Vosshall LB (2013). orco mutant mosquitoes lose strong preference for humans and are not repelled by volatile DEET. Nature, 498 (7455), 487-91 PMID: 23719379

Stanczyk NM, Brookfield JF, Field LM, & Logan JG (2013). Aedes aegypti mosquitoes exhibit decreased repellency by DEET following previous exposure. PloS one, 8 (2) PMID: 23437043

Klun JA, Kramer M, & Debboun M (2013). Four simple stimuli that induce host-seeking and blood-feeding behaviors in two mosquito species, with a clue to DEET's mode of action. Journal of vector ecology : journal of the Society for Vector Ecology, 38 (1), 143-53 PMID: 23701619