Research Methods: The Path of Least Resistance

Friends, family, and colleagues,

Forgive me for not writing sooner. I’m wrapping up the last of my experiments and unfortunately, that must take priority over blogging.

Measuring Pathogenicity

The last test I’ve been conducting is a test for pathogenicity. Pathogenicity is defined as an organism’s basic ability to parasitize a susceptible host. This can be broken down further by an organism’s “virulence” which is often characterized by having certain genes for enzyme or toxin production that makes it even better at parasitizing. For this test, we aren’t going that in-depth with our tests.

The Disease Triangle

Before I explain the experimental setup, I need to back up and explain an important concept in plant pathology, the disease triangle.

The triangle, as one would expect, consists of three sides. Each of these sides represents an aspect of a successful infection. In order to have infection, all three of the following aspects must be present.

Courtesy APS

First, a virulent pathogen must be present. This goes without saying, but if the pathogen isn’t present, then there is no disease. Second, a susceptible host must be present. Finally, the environment must be favorable for the pathogen to infect.

The disease triangle, while simple in its design, is incredible versatile for plant pathology research. Keeping two of the sides fixed allows researchers to conduct various experiments in a simple, straightforward manner.

For example, a plant breeder may screen for resistance by keeping the pathogen and favorable environment fixed which leaves the breeding lines to be the variable. Still another researcher interested in a fungal population can fix the host and the conditions and screen the population to gain insight on how pathogenic the population is on a susceptible host. These are simple examples and the variables that are changes can be much more subtle than what’s described here. The point is that the possibilities are nearly limitless.

If this disease triangle really interests you, I'd recommend visiting the APS website.

The detached leaf

For this experiment, I’m using a method called the detached leaf assay. Just as it sounds, it involves a leaf that has been detached from the plant. This method is fast, consistent and takes up very little space. Originally, the plan was to do this and a seedling test. However, the lab I’m in is not quite right for seedling tests. The growth chamber I’m in is set up for a process called tissue culture. So it was decided that a closed system like this would work better.

The leaf is exised (cut) from the plant and then is dipped in 75% ethanol to kill any insect pests. After a few minutes air-drying, the leaf is placed on a piece of filter paper which is then placed inside a glass petri dish. The paper is wetted with a couple milliliters of water and the pathogen is transferred from an agar plate. The petri dish is then sealed and measured at intervals of 48 hours for 6 days.

"Where are the controls?"

Whenever looking at an experiment, one question that scientists often ask is “what were the controls”. Without controls, there is nothing to compare the treatments to. In this experiment, I started out with a simple control, an untreated leaf. I followed all the instructions, except adding the pathogen. This allows me to see the effect that cutting the leaf and placing it into the chamber will have. A second control I used included a blank agar plug on an untreated leaf. This allows me to see any effect that the agar or anything in the agar may have on the leaf. Without these controls, it would be difficult to draw any useful conclusions from the data collected.


Rating system

The rating system is simply a 0 to 10 scale based on how much of the leaf tissue is necrotic (dead). A 0 is typical for our controls, as there’s nothing to kill the leaf and 10 is a really pathogenic isolate! I’ve added some photos so you see what some of the results look like.

No Pathogen? No problem!-Untreated plate (6 days)

Weakly pathogenic isolate (6 days)

Moderately pathogenic isolate (6 days)

Highly pathogenic isolate (6 days)

Quick Update-I'm coming home (soon).

I'm only here one more week. So I should be able to post a little more and catch up on a few more of my adventures while I've been in Nanjing. Stay tuned.

Mason

Research methods: Resistance is futile

Friends, family, and colleagues,

This post will discuss the main topic of my dissertation, fungicide resistance. To bring you up to speed on my pathogen, check out this publication

What is fungicide resistance?

Fungicide resistance or fungicide insensitivity is the phenomena where fungi can become less sensitive to fungicides after repeated exposure. In its simplest terms, when a fungal population (let’s say a population of Didymella bryoniae, my pathogen) is exposed to a fungicide, most of the fungi die off. However, as there is variation in the population and some of those variants have mutations that allow them to grow and reproduce in the presence of said fungicide.

If this sounds like something you’d read in Darwin’s “On the Origin of Species”, it’s because the process of fungicide resistance is a form of selection.

Why is fungicide resistance worth studying?

With many crops, there are multiple tools to combat pathogens. Growers can grow resistant cultivars, in which the plants are less susceptible to the pathogen. Growers can change the planting date in some cases to avoid the pathogen. They can also use treated seed to reduce the initial pathogen load.  With watermelon, there is no commercial resistance available. Growers in Florida can grow early, which helps avoid some pathogens. Seeds are regularly treated, but problems still persist.

The primary way that growers manage gummy stem blight is through the use of fungicides. This is problematic as fungicides can be expensive and resistance has been reported. It is important to characterize populations in the major growing regions to determine how much of a problem it really is.

So, why did you go to China to do this work?

We live in a global economy and 22% of all the watermelon seed imported into the US comes from China. In fact, if we look at the top 5 nations that export watermelon seed into the US, it accounts for nearly ¾ of all the seeds imported. The other four countries are Egypt (21%), Chile (17%), Peru (9%), and Israel (5%).

Given that D. bryoniae can be transported long distances on the seed, it is worth exploring potential risks of selecting for fungicide resistance in seed production areas as well as commercial production areas.

How is fungicide resistance measured?

There are many ways to measure fungicide resistance. For the purpose of my trip, we use a simple method called an amended agar assay. We grow the fungus on a nonamended plate and then we take a 7mm agar plug from that plate and place it on an amended plate.

All my agar plugs are placed face down so the fungi has to grow on the new plate

My fungus grows well on a media called Potato Dextrose agar, or PDA. Amended agar is exactly what it sounds; it is amending the agar with the fungicide. For this test, we add a specific amount of a single fungicide and compare its growth with an unamended fungicide.

There’s a lot of ways to measure, but I prefer a simple method. Typically, fungi will grow in a radial pattern, meaning that it will grow out from the center in a circle. I measure two points that are perpendicular (90^o from each other) at three time points. The time points for this fungus that work
best are 48, 72, and 96 hours. I’m very fortunate to have a fast growing fungus.

Control plate (right) with my 5 fungicide plates (center).
The top plates incidcate the this isolate is resistant to those fungicides while being sensitive to the bottom.

How much fungicide is used?

The rates of each fungicide that we’re testing varies. Fortunately, research has been completed to figure out a “baseline sensitivity”. The Fungicide Resistance Action Committee (FRAC) defines baseline sensitivity as:

“A profile of the sensitivity of the target fungus to the fungicide constructed by using biological or molecular biological techniques to assess the response of previously unexposed fungal individuals or populations to the fungicide.” 

This is simply how sensitive a population is to a specific fungicide. It has to be done for every fungicide I wish to test in order for my method to work. If it hasn’t been done, I get to calculate the baseline EC₅₀ with a lot of doses of the fungicide_. In my methodology, the fungal population is assumed to be sensitive to the fungicide at the EC₅₀. This is my discriminatory dose.

What is EC₅₀?

The EC₅₀ indicates the dose required to inhibit growth by 50%. So, for an isolate sensitive to a fungicide, say boscalid, the EC₅₀ is about 0.0034 mg/L. For every liter of agar I pour, there is 0.0034 milligrams of active ingredient in that agar. If the fungus is sensitive, it still grows (usually), but the growth is much smaller compared to a petri dish without the fungicide.

To give you an example of how small this is, take a typical 81mg aspirin tablet. Take that tablet and cut it in half. Repeat this halving for 10 more times. Now you have about 0.0034mg to put into a liter of agar.

I hope this was clear enough. If you have specific questions about fungicide resistance, feel free to leave them in the comments section. There are many different setups and I’ve given you an oversimplified method of what I am doing.

Mason

Watermelon industry day

Friends, Family, and Colleagues,

A few weeks ago, JAAS hosted an industry day for the watermelon industry in the Jiangsu province. It was similar to any industry convention you'd go to in the US, I've decided to make this a picture-heavy post to give you an idea of the varieties that are being produced on the other side of the world.

There were a LOT of free samples. It was mostly watermelon, but they did have muskmelon (which they call sweet melon over here).

Banner at the entrance

Opening remarks from the coordinators

Not all the watermelons were round...

There was a good turnout. Note the little kid pointing in my direction.
Americans are uncommon in this region of Nanjing.

This variety is translated to "ice cream" in English. Note the yellow and orange flesh. It was very sweet.

This was one of the trellis exhibits in the back of the building.
I'm here with Yuan, one of the students.

There is a tomato industry day this Saturday that I may check out. I don't know if there will be as many free samples. However, I could be mistaken, the Chinese love their tomatoes.

All is well over here.

Mason

Research Methods: Single Spore Isolation

Friends, family, and colleagues,

As part of my blog, I’d like to share with you, my readers, what I’m actually doing now that I’m settled into the institute. For the sake of my research, I won’t be discussing any results. That’s for once the seminars begin. For the sake of your sanity, I’m also keeping the methods very general, since I want this to reach to everyone. 

In plant pathology, it is said that the journey of a thousand experiments begins with a single spore. Ok, no one in my field says that, I just needed a cheesy introduction. 

The first step when I arrived to JAAS was to take the collection of isolates and culture them from a single spore. This technique is appropriately called the single spore isolation. This is done with most fungi for various reasons. The most important is uniformity. If you have a pure culture (that is, only one organism growing in your plate), there is still no way to ensure that the one organism is a single lineage or if it’s a diverse population. You could have 15 different individuals of the same organism in there, and that could cause some inconsistencies with experiments. Sometimes the fungi is grey or white, next time it’s green! 

This is NOT a pure culture. Some contaminants are clearly present

This is a pure culture. There's only one organism growing in there.
But is it all the same organism or a population of the same organism?

First step: Get spores

It may surprise you that in order to do a single spore isolation, you need spores. Now, that can be challenging for some fungi, since they don’t make spores. I had a labmate who worked on a pathogen called Sclerotium rolfsii that does not produce spores. There are some methods that can be used if that happens. However, that’s a whole different topic that I’m glad to avoid.

For my fungus, it produces two types of spores, asexual (called conidia) and sexual (called ascospores). Each kind of these spores is produced in a separate structure. Conidia are borne on a structure called a pycnidia, whereas ascospores are borne on either a perithecia or a pseudothecia (don’t get too hung up on the names, it has to do with the shape and structure of the fruiting body). For the sake of simplicity, conidia are the ones I want.

This a perithecium with ascospores emerging.

Pycnidia with conidia being released.
Each spore is a clonal copy of its parent

So how do I get these conidia?

The fungus has to be producing these pycnidia in order to get spores. There are a few methods that I use to get said spores. First, adding light to the growth chamber encourages the fungus to grow. Just as plants respond to light, so too will fungi. It isn’t essential for their growth, but it triggers certain responses, including pycnidiation. Adding in UV light also encourages pycnidia to form. Another method is to grow the fungus on a minimal media. If the fungus runs out of food, the next step is to make spores to find a better environment. 

I have spores! So now what?

So once we have spores, we have to count them. The best way is to take some of the spores from a petri plate and scrape them into some sterile water. At high enough concentrations, you can actually measure optical density (i.e. how “blurry” the spores are in water). However, there’s another way if fungus doesn’t produce billions of spores. There is a device called a hemocytometer (sometimes spelled with an "a" hemacytometer). Originally, it was produced to count red blood cells. It is also used to do sperm counts in both animals and humans. Fellas, if you ever go in for fertility testing, there’s a good chance that one of these will be used to assess your sperm.

Thanks Wikipedia!

So, the spore suspension is placed on the hemocytometer, and on the surface, there is a grid pattern. That grid pattern is viewed under a compound microscope. Within each grid is a known volume, typically 0.1µl. That’s one microliter, take your liter of cola and divide it by one million, Farva. We count the number of spores and from that concentration, we can adjust out suspension of spores to a desired amount. I’ll spare you all the math.

A typical grid pattern on a hemacytometer,
The red area (letter A for color blind) is 0.1ul in volume

I did the math! Now what do I do?

Once the spore suspension has been adjusted the suspension is then dropped onto another petri plate. For the sake of simplicity, I’ll go through an example. Say the suspension is adjusted to 1000 spores per 1ml of water, each one of those little droplets is going to be 1µl, which is 1/1000 of 1ml. In theory, there should be 1 spore within that aliquot. This is repeated several times as there is a lot of variance within that suspension. The conidia often come out of the pycnidia in a ribbon and are held together by a gel-like substance; adding a detergent reduces the gelling. Often, it’s called a “spore horn”. Also, this assay does not distinguish between viable cells and dead ones, so more chances are better chances. After the suspension has been dropped, the plate is left alone for 24 hours. 

If you mess with the spores, you sometimes get the horns.

I waited 24 hours, what’s the next step?

After the 24 hours, the plate is observed under the microscope and the spores will have germinated. Now comes the tricky part, an aliquot containing exactly one spore must be found. Given the variance, it’s not uncommon to find one aliquot with 6 germinated spores and the next aliquot to have none. After a bit of patience and luck, one perfect aliquot is found! We use a cork borer to carefully cut the agar chunk out of the original plate and that chunk is then plated onto a fresh, clean plate. After 48 hours, the single spore will have grown to the size where it is detectable to the naked eye. 

I successfully single-spored, now what do I do with it?

Now you can run all of your experiments! 

This is only the first step in the research journey. From start to finish it takes about 3 days to complete, assuming the culture is already creating spores. For the sake of consistency, nothing can be done before the single spore isolation, so it becomes a crucial task if working with fungi. I plan on generally describing some of the other experiments I’m running in the lab. I know I used a lot of technical terms, so feel free to ask for clarification in the comments.

I hope you found this interesting. If you have questions, leave them in the comments.

Mason

Top ten: things that are common in the US, but are rare in China

 Friends, family, and colleages,

In an effort to make a few shorter posts for those who don't have all day to read my ramblings on China, I have a series of top-ten lists.

1. Social media: the Chinese government has blocked most social media, including Facebook, twitter and Instagram. I think it has something to do with cutting down on dissention/organized protests. I use the VPN to get around this.
2. Google: all things google are blocked. Which makes it difficult, since my default email is powered by google. Again, the VPN is useful in this regard.
3. Cheese: In the US, we smother a lot of dishes in cheese. Here, I’ve had one or two dishes with cheese (one of which was a pizza shop). Cheese in the markets are very expensive as well.
4. Water fountains: If you drink water all day like most Americans, you’ll have a hard time adjusting to China. Though nearly every office has a water boiler for tea.
5. Wifi: In my office, the only way I can get WiFi is to plug in a little device into my laptop and it turns computer into a mini-router. There are no dedicated routers in my office, though there are routers in the mall
6. Refrigerators: In the US, there’s one in nearly all the student offices, plus one in the break room. Refrigeration is rare except for essential activities. I don’t know about individual apartments.
7. Soda: It’s available in some stores, but it isn’t nearly as ubiquitous as it is in the US. It’s rarely served in restaurants here.
8. Gasoline-powered scooters: Both at UF and China, there are plenty of scooters, but most of the scooters here are electric (and subsequently very stealthy, look both ways before crossing the street!)
9. Silverware: chopsticks for days!
10. Coffee: In China, green tea is much more popular. As a result, I haven’t had a cup of coffee in several weeks. There are a few coffee-shops in the mall, but the prices are about the same as a starbucks in the US.

In China, they eat pizza with their hands, but they wear gloves to keep their hands clean.

Mason

Energy and Development

Friends, family, and colleagues,

First, let me apologize for not posting in a bit. To say I've been busy is an understatement. I've been making a lot of progress with the research, which is good. After all, that's the primary reason I'm here.

For this post, I want to share my observations on two topics that go hand in hand; energy and development.

Energy:

Energy is used slightly differently in China compared to the US. The resource isn’t scarce, but there are a lot of people. Air conditioning is rarely used. Refrigeration is used for essential functions. We have two coolers in the three labs we share here. There are no mini-fridges in the offices. Water is served hot (for safety) and they don’t see a need to cool it down afterwards. Public transportation is encouraged and is very affordable. I took the bus the other day for 2 RMB (about 33 cents American).

Green/alternative/renewable energy is a big topic both here and in the US. However, the questions they ask are entirely different. In the US, the typical question is “How can we use renewable energy to make more electricity?” In China, however, the questions are often “How can we use renewable energy to use less electricity?” In China, energy conservation is heavily encouraged. From what I could gather from the energy conservation is also pitched to the people as good citizenship/patriotism. That isn’t necessarily a bad thing, though I don’t know how that would be received in western cultures. Most of the energy conservation is pitched as a saving money on bills.

Take for example hot water. In both the US and China, each home typically has a water heater, the purpose of this device is to heat water and make showers bearable (among other uses). In my household growing up, this was in the basement. I’m guessing it was placed there to be close to the water pump. In the US, you can typically buy an electric or a gas powered heater. Now, in China, the hot water situation is a bit different. It is much more common to find a solar heater, which uses the power of the sun to heat the water. These are found on the roof of buildings in nearly every household. I have a photo of one posted below. (Note: These are broad observations and I’m sure there are exceptions in both cases.)

A typical solar heater in China.
While these are available in the US, they're not nearly as prevalent as they are here.

Development:

China is growing fast. When I say fast, I mean as fast as possible. Large cranes are a common sight in every city. When a building is completed, move-in is immediate. The institute is in a brand new building, and there is still construction dust on the door frames and wrappers on the door handles. My dorm room just got the screens last night and the guy installed them in seconds and quickly left to install the next ones. The packing tape was still attached. There is an intense demand for building space and housing, which contributes to the boom in construction.

It’s not uncommon to see multiple cranes going in a building complex. I have probably seen over 200 cranes since I’ve been here. I’m told that large trucks are not allowed to deliver materials during the day. At night, however, numerous trucks can be seen on the city streets (and even more so on the city’s highways). I’ve also been reading reports that Chinese development is slumping as of late.

From what I can ascertain, a lot of the buildings being erected are apartment complexes. Housing is in extremely short supply. This shortage has resulted in very high housing prices. An older flat in Nanjing is priced around $400,000. As a result, a lot of the people rent, which, at 3,000RMB a month, it isn’t cheap either.

Even in the slump, the growth rate of China is astronomical!

Hope all is well. I will be home soon! The Wi-Fi at the mall is really buggy, so I apologize for not posting more photos. I'll try to post tomorrow when there are fewer people on the routers.

Mason

Culture Shock: Chicken fingers?

Friends, family, and colleagues, 

These posts titled "Culture shock" will include items that are starkly different from something we would see in the US. While I'm not necessarily shocked by them, I notice that they are very unusual compared to what we would normally see in everyday America. I hope you find these amusing and entertaining.

So, one of the students offered me a snack today. Not wanting to be rude, I accepted it without hesitating. I bit into it and noticed that it was salty, yet spicy. Only then did I look down to see what I was eating. 

It still has the claw. I guess so you can pick your teeth afterwards.

When I was a little kid, we used to slaughter the extra roosters and eat them. When I was 8 years old, I thought it would be so cool to keep all the chicken feet. So, my father being the loving man he is, gave me all the feet, which I kept in a paper bag for well over 2 months. Looking back, that was really gross. Even then, I would never think about eating them. 

So, not being rude, I kept eating it. The texture is about what you'd expect, rubbery and chewy. I didn't eat it whole, rather I just gnawed on it for a while. At least it was seasoned well. 

Comfort zones are overrated anyway.

Mason

Cooperative Extension Part 2

Friends, family, and colleagues,

This is the continuation of our outing in Dongtai. You can find part 1 on a previous post.

After we talked to the grower at Dongtai, we traveled to another operation with a “new disease” in their operation. As extension folks, we love disease, but to really get an extension pathologist excited, use the words “new disease”. The symptoms were described as a swelling at the base of the rootstock plant and wilting of the scion plant. This was a problem only on the grafted watermelon, though his nongrafted melons had wilting as well. While we drove to the field, Dr. Paret and I speculated on what we might find. Is it a bacterium? A virus? Herbicide damage?

Typical operation in Dongtai, Jiangsu Province. Melons are grown under protected structures.

Grafting vegetables

As I mentioned in an earlier post, nearly all the watermelon in China is grafted. To graft a watermelon, the seedling is cut and then reattached onto a closely related species (gourd or pumpkin usually). The top portion (the watermelon) is called the scion while the bottom portion is called the rootstock. This is very popular in the US for tree fruit production to dwarf the plants to make harvesting easier. There is a lot of utility in grafting one plant onto another. You may find more information here.

The purpose of grafting in this context is to maintain resistance to a pathogen Fusarium oxysporum. This is a soilborne pathogen that infects the roots and colonizes the vascular tissue. The fungus, coupled with the host response, wilts the plant. The disease is appropriately called Fusarium wilt. However, Fusarium wilt is very host specific. Each species can be broken down further by forma speciales, which is latin for form species. This explains what the host of the pathogen is. For example, the forma speciales that goes to watermelon (Fusarium oxysporum f.sp. niveum) cannot infect closely related species, such as pumpkin or squash. So, if you have a problem with Fusarium wilt in your watermelon field, one solution is to graft your watermelon onto a pumpkin, gourd, or squash.

A grafted melon with the graft union in blue.
There was also some gummy exudation (red), which is a classic symptom of my disease

Now here is where it gets (disease) complex

However, Fusarium oxysporum often has help in infecting plants. A small roundworm, called a nematode lives in the soil and feeds on the root of plants. These create wounds which are exploited by the Fusarium to cause infection. This is known in my field as a disease complex, where the disease is more severe in the presence of another pathogen. Unlike the Fusarium, nematodes have very broad host ranges. They are not picky eaters. They can attack the roots of squash, pumpkin, gourd, watermelon and many more. By itself, nematodes can cause a lot of damage to crop roots resulting in wilting and stunted plants.

Investigative plant pathology! Crop Scene Investigation

I think you know where this is going. We get to the field and the grower points to one of the symptomatic plants. The rootstock is fat, but the scion is wilted and dying. Dr. Paret pulls up the plant and the first thing I notice are galls on the root. We have found our first clue. The roots are sparse and covered in swollen galls, which are characteristic of nematodes. When the nematodes feed, the roots respond by dividing their cells rapidly and expanding the cells they already have (known as hyperplasia and hypertrophy respectively). 

Courtesy: http://www.expert-nutrition.com/muscle-hypertrophy.html

Looking for more clues

The next step to a diagnosis is to cut the stem open with a knife to check the vascular tissue. We find that the grafted plants are clean. There is no visible discoloration which we would see if there was Fusarium wilt. The swelling was caused by the few roots left trying to push the water up, but not being able to make it past the graft union. The scion wilted while the rootstock swelled. How cool is that?!

Galling caused by root knot nematode

Stem swelling. Note the tiny amount of roots.
Notice that there is no discoloration in the vascular tissue.
This plant is not infected with Fusarium wilt.

The second “crime scene”

Next, the grower takes us to some non-grafted plants. He thought that maybe the rootstocks were defective, but the same thing was happening. We pull one up, with only a few roots (lots of nematode damage). We cut open the stem to be met by a brilliant orange discoloration! You have Fusarium wilt! Ok, we weren’t THAT excited, but it was close.

Classical symptoms of Fusarium wilt. Note the orange discoloration along the vascular tissue.

Delivering the news

We explain (though a translator, I don’t know the word for nematode in Chinese) about the problems this grower has and some possible solutions. He peppers us with many questions about rotating fields, what this means for his production, et cetera. The cool part about cooperative extension is because the grower is a captive audience, he is truly listening to what we have to say. What’s even cooler is his 17-year-old son is listening even more intently (and taking LOTS of photos of the symptoms), probably because he will someday inherit this farm and wants to know what problems he will face.

It is experiences like this one that make it so rewarding to work in the applied sciences. I think all sciences have merit and every finding benefits mankind in some way, shape, or form. The extension work we do isn’t simply cataloged in a journal that only a few academics will read. It’s disseminated into real world situations. So while we didn’t find any new disease, we did help a grower solve some problems and educated him (and his son) on possible solutions. That’s what I love about my job!

Dr. Paret explaining the symptoms which is then translated via Dr. Ren (grey shirt).

I also think it's amazing that you can travel thousands of miles to find the exact same problems that you see in your own home. Experiences like this one are extremely rewarding. Who knew two guys from Florida would find the same pathogens a world away?

Mason

Cooperative Extension Part 1

Friends, family, and colleagues,

Last week, we had the opportunity to do some collecting and that involved going out in the field. In academia, a lot of time is spent in an office answering phone calls, responding to email, and other boring office tasks. However, there are times when they do let us out to play. This is an extension specialist’s favorite activity (or at it’s least my favorite activity anyway). Field visits are an opportunity to take what we have seen in a book or in a presentation and apply it to a grower’s situation. 

"A pathologist is the only person who is excited to see a disease in the field"-Mathews L. Paret

What is Cooperative Extension?

Cooperative extension is a unique wing of academic research. It is an integral component of any college of agriculture in the US. The overall purpose of extension is to take the information gained at academic institutions and to disseminate said information to the public. For my field, that means taking the knowledge I gain from my research and applying it to watermelon growers’ operations. Cooperative extension was established over 100 years ago with the Smith-Lever act of 1914. 

Office Visit

This day, we were out in Dongtai, Jiangsu province. We started out talking to some staff at the ministry of agriculture. From what I could gather, they appeared to be a provincial level governmental office. (think Florida Department of Agriculture and Consumer Services-FDACS or the Delaware Department of Agriculture-DDA). Their job was mostly policy and grower support from what I gathered. None of them spoke English, so we got most of our information through a translator. 

Ministry of Agriculture building. There were three more buildings surrounding it.

Field Visit

The grower was harvesting one of the institute's varieties "Sumi #8", it's a sweet melon weighing about 4 kilograms with yellow flesh.

After a lengthy meeting at the ministry of agriculture, we drove out to the first operation. They were spraying fungicide, insecticide, miticide, and what appeared to be a foliar fertilizer in a 55-gallon cocktail mix. Their philosophy was to spray for everything just in case. This is not only wasteful and environmentally harmful, but fungicide resistance can develop if products aren’t rotated properly.

Here's the tank-mix of all the chemicals.
Not quite as advanced as the sprayers we use at the university.

This is a copper compound used for control of bacteria and fungi.
Many bacteria are tolerant/resistant to copper

What is Fungicide Resistance?

One phenomena of the diseases I work on is that repeated exposure to the same fungicide time and time again can select for resistance. This means that the sensitive isolates are controlled, but the resistant isolates keep growing without any inhibition. If you’re a fungus that is resistant, this means that you will continue to grow and reproduce. This is problematic for the grower since 1) it’s expensive to use something that doesn’t control the disease and 2) it can cause problems for other growers or the same grower in other years. The collective term is fungicide resistance or fungicide insensitivity. This is a serious problem in hospitals with “superbugs” and insect pests, which is caused by the same phenomena. 

This is Imidacloprid, an insecticide, mixed with another insecticide.
It is the compound often blamed for honeybee decline in the US.
Some species of insects are also reported to be resistant.

Extension in action: Delivering Recommendations and Sampling the Goods

We talked to the grower for a bit about what he was using and we discussed the importance of rotating fungicides and to use only fungicides for control of gummy stem blight. He was also using some antibiotics, which we explained won’t control gummy at all and could cause problems down the production line. He was very grateful for the advice and we ate lots of watermelon in his make-shift home.

Dr. Paret with the grower and some ministry of agriculture officials. Notice the table, that's the remains of two watermelons and one cantaloupe after we ate. They were sweet as candy!

I’m making this a 2 part series, since this day was really neat with a lot of extension opportunities. I’ll post the second part soon.

Mason

Small World: Coca-Cola

Friends, family, and colleagues,

In this series "small world", I will attempt to bridge the massive cultural and geographic divide readers see in the "culture shock" series with facets that can be used to relate to one another. I will post similarities we share that can be used to show that even a world away, some things are still familiar to us.

On Saturday, we went to the Nanjing Museum. Some of the students missed breakfast, so we stopped at the supermarket to grab a bit to eat. One of them bought me a coke. It was a welcome sight!

The bottle has the trademark Coca-Cola signature sketched into the plastic. The characteristic red label was written almost completely in Chinese. The bottle itself was a bit taller than the typical US bottle. The volume was comparable to the US at 600ml (20.29 fl oz). The flavor was very similar, though the sugar content seemed to be a bit lower than in the US, which is typical of most things found in China. I wasn't able to read the calorie content, since the energy was measured in KiloJoules (KJ) instead of calories.

Mason

Top Ten: Tips for traveling to China

Friends, family, and colleages,

In an effort to make a few shorter posts for those who don't have all day to read my ramblings on China, I have a series of top-ten lists.

1. Be patient-The flights over here totaled about 26 hours from Gainesville, Florida. Make sure to bring a book (or 2 or 3) and a podcast or two.
2. Try new food- This is not the time to be a picky eater. In addition to lots of rice, there's a lot of meat and fish dishes to try. I was never a big fan of fish, but I got over that very quickly, mainly out of necessity. 
3. Don't be a germaphobe- Your chopsticks will likely pick food out of the same bowl as everyone else. Just make sure you're up to date on your Hepatitis B vaccinations and you'll be fine.
4. Have an English-speaking friend-This makes traveling and going to the market easier.
5. Bring plenty of deodorant- In the summer, you will sweat a lot and deodorant isn't very common in markets.
6. Bring cash to exchange-Even if your credit card is cleared for international travel, it doesn't necessarily mean that banks will accept it. Cash is still king in China.
7. Make sure you're vaccinated-Check the CDC website before you go and make sure you have all the vaccinations. One preventable disease is enough to side-line your entire trip and put you in real danger.
8. Learn a few phrases in Chinese-Even though a lot of Chinese people speak a little English, a few phrases will be helpful (and at the very least, make the locals chuckle).
9. Set up a VPN-This will allow you to connect to a virtual private network in the US. The great firewall is a thing, and it blocks a lot of sites including this blog and Facebook. The VPN allows you to tunnel though the firewall and connect to "blocked" sites.
10. Have Fun-Take lots of photos and explore a lot. There's a lot of amazing things just in the city where I am staying. 

I found some English-speaking friends quickly at JAAS. They are very welcoming.

I hope these tips will be interesting and useful to anyone planning on traveling abroad. Generally, speaking, I think these tips could be applied to any country for any trip. 

Mason

First impressions; safety, spitting, and selfies

 Friends, family, and colleagues,

So, I’ve been here over a week and I’ve had some time to make more observations on my experience so far in China.

Safety.

I have felt completely safe during my time here. As I mentioned in an earlier post. Security is overt but not overbearing. They are there to keep everyone safe, especially foreigners. The campus also has a gate-keeper and a series of guards at the front gate. They aren't there to intimidate people, just to keep everyone safe. They’re always friendly to me when I enter or exit. Some of them have tried to practice English with me. Probably the most dangerous part of the day is crossing the street. Vehicle lanes are largely suggestions in the city and even more so in the rural parts. To add to the complexity, the city also has moped scooters and bicycles in separate lanes (which don’t always stop at stop-lights). I've found that following the locals is the best way to go. Even with the crazy driving, there are surprisingly few accidents. I've been here 10 days and I've seen 1 accident (a small fender bender between two cars) even with the hundreds of kilometers traveled.

Not sure if you'll make the light, they'll let you know how long you have...

...And how long that light is going to be red

For every car in the city, there are about 2 or 3 scooters

Spitting

Spitting is fairly common in the city streets. I’ve also seen my fair share of nose blowing (see snot-rockets). That’s about as much time as I’m comfortable spending on this topic (I needed another S-word for the alliteriation). Closed-toe shoes and staying off the grass are good advice.

Selfies

Selfies are very popular in the US and are even more popular in China. I've seen more selfie sticks than I can count in the first week I've been here. I don’t know where all these photos are going (remember that social media is blocked), but they love taking photos. I was afraid that I would look like a tourist taking all these photos, but then I realized that everyone else was doing the same thing. I've also seen several folks with spare battery packs on their phones for added power.

We saw this girl at the Mausoleum.
She must have taken over 200 photos of herself

Just like people, selfie sticks come in all shapes and sizes

So, for those of you who want the cliff-notes version:

1. China is incredibly safe. 
2. People like to spit in China, watch your step!
3. Chinese love taking selfies.

Mason

The party is over, get back to work!

Friends, family, and colleagues,

Dr. Paret has gone back to the US and I've moved from the hotel to the dorm room. It’s a step down from all the showcasing that I saw last week with the flashy meals and excessive amounts of food. I expected this much, this isn't exactly a vacation!

Dr. Paret showing one of the scientists classical symptoms of root knot nematode (RKN) on a watermelon rootstock

The dorm room is nice, especially when compared to some of the older apartments in the area. It’s mostly furnished. It does have a bed frame (no mattress), and a cabinet dresser. From the sound of it, mattresses really aren't a thing here. There is also a stool and a desk along with a bathroom (with a sit-down toilet thankfully!). It was a bit dusty when I moved in, which is a symptom of being brand new and no one living in it for a while. I tried mopping, which helped a little bit. The room itself is very quiet and has that strange echo sound you get whenever you move into or out of a completely empty apartment.

The dorm was built in 2012, and is one of the nicest housing options available to students at JAAS.

There's a lot of little sitting areas outside on the campus

Some students live in the old apartments across the pond. Due to housing shortages, to buy even an old apartment is very expensive

The academy itself is a very large campus, it houses about 2300 employees. Most of the graduate students and faculty live on site. The main road is lined with what I think are sycamore trees. Lunch which used to be never-ending has now been replaced by three meals from the academy’s cafeteria. It’s nothing close to the food I had a week ago, but it will have to make due. You cannot beat the price though. I ate breakfast for 2 Yuan, about 33 cents American. They had these Chinese onion pancakes that are pretty good for institutionalized food. Lunch and dinner run about 8 to 10 Yuan ($1.30-$1.60).

The labs are well stocked by Chinese standards. We definitely produce a lot of waste compared to the Chinese labs.  The sizes of the Chinese and the size of this American are comically incompatible. I had on a pair of small gloves and a medium lab-coat and Dr. Li couldn't stop laughing. I’m pretty sure this is what a T-Rex would feel like if they worked in a lab. Labeling items is a challenge, as they don’t have any of those tape dispensers in all of our labs. Fortunately, the sharpie marks come off with ethanol. They have a very small autoclave in the lab. It works well enough for bottles and small items. It's also nice to have my own little autoclave in the lab.

In addition to the dorm and the labs, they also have a graduate student lounge and pingpong table. I’m sure I’ll eventually get roped into a game. The students here are very welcoming. Just as in the US, there’s a lot of young women in the field of STEM. They spend most of their waking hours either in the laboratory or in the fifth floor reading center. 

All in all, I'm settling in and meeting people. I’ll try to explore the area. So far, the only place I've been is the forest mall across the street. It’s a modern mall with a lot of free WiFi which lets me use the VPN and talk to you!

Mason