Measuring non-photochemical quenching in a few seconds without an initial long dark acclimation.
Over the past 3 years, many MultispeQ users have noticed that the NPQ(T) parameter (and ΦNPQ) can be a powerful predictor of plant stress, either biotic or abiotic. The NPQ(T) parameter has also correlated with crop yields in some PhotosynQ projects, like this project from Malawi.
Indeed, one of the big breakthroughs with the MultispeQ is the ability to estimate NPQ (Non-Photochemical Quenching) without a long dark acclimation period, which allows us to develop robust protocols that take less than 20 seconds. So how is the NPQ(T) parameter derived and how does it compare to the established NPQ parameter?
Tietz et al. out of the Kramer Lab have just published a paper in Plant, Cell and Environment describing the parameter and its derivation. Congratulations!
Tietz, S., Hall, C. C., Cruz, J. A., Kramer, D. M. (2017) NPQ(T): a chlorophyll fluorescence parameter for rapid estimation and imaging of non-photochemical quenching of excitons in photosystem-II-associated antenna complexesPlant. Cell Environ. 40(8), 1243–1255. doi:10.1111/pce.12924
The first version of the MultispeQ, the MultispeQ beta has been a great instrument, workhorse and proof of concept for the PhotosynQ platform and its utility in phenotyping plants outside the lab in large sample sizes. There are still MultispeQ beta instruments out there which are in use. We have decided with a heavy heart to stop the active development for them, since they have already exceeded their anticipated lifespan and focus our limited resources on the new MultispeQ v1.0.
So, what does it mean?
You will still be able to use the instruments and use the existing protocols, as well as create your own new ones. If there will be a change that breaks the compatibility with the Platform, we will give you enough of a heads up, so you can finish your experiments before we release the update. Since all the informations about the instrument is open, we hope that fixes or improvements might be made by the community to extend the instrument’s lifetime.
We are no longer supporting hardware fixes, mainly, because we don’t have parts in stock any more. Pieces like the light guides were custom made and can’t be ordered. Electronic parts can be ordered and we are more than happy to point you to where to source the needed parts. Just let us know and write to firstname.lastname@example.org. Otherwise, we advice you to get the new MultispeQ v1.0 for your future data collection.
There are no more measurement protocols developed for the old MultispeQ beta. The structure and some of the commands have changed when we introduced the new MultispeQ v1.0 and some of the new features would need a complete re-write of the instruments software (firmware).
We are no longer updating the firmware since we want to focus our limited resources on the current instruments and make sure they receive updates and improvements on a regular bases. As long as the communication protocol doesn’t change, the instruments can be used with the apps and submit data to the PhotosynQ platform.
We would like to say thank you again to all the beta testers not only for testing the MultipseQ beta, but the PhotosynQ platform as a whole. We learned a lot and the new MultispeQ v 1.0 has benefitted from those experiences a lot.
Does it seem like your MultispeQ measurements are taking too long? You heard each measurement should take 15 seconds but you feel like it is taking almost a minute? There is a simple and easy solution!
One of the most common issues that users of the MultispeQ v1.0 are reporting is that their measurement time has dramatically increased when using the “Leaf Photosynthesis MultispeQ v1.0” protocol, taking up a minute to finish. What the heck is going on here, are all these devices lagging or broken? No! In fact this issue is due to a new command we introduced into the protocol.
Before, when you wanted to take a measurement in the field, you would probably answer project questions on your phone, clamp the leaf using your MultispeQ, and then hit measure in the app. It would take 15 seconds, and everything was great, so why did we change this order of operations? Well, there was always a delay from when you hit measure to when the actinic lights turned on, during which time the leaf begins to dark adapt.
Now, we have added an open/close start command that it is really cool and will hopefully make your measurements easier. The open/Close start is now at the beginning of the protocol code, because it happens before anything else. Utilizing the magnet in the MultispeQ, we are able to detect when the devices clamp is fully open, and when it closes. Using this, users are now able to fill out their project questions, hit measure, find their leaf, and then clamp the leaf just like you normally would. The difference? Now the actinic light turns on, and the measurement starts as soon as the MultispeQ senses that the clamp is fully closed.
Another advantage of this feature is that it provides you with the option to hit measure, set the phone down, and use two hands to make sure you properly clamp the leaf. This can be especially useful when it is difficult to properly clamp the leaf, such as with small leaves.
If the device does not detect that the clamp fully opened and then closed, it will wait for 30 seconds and then automatically start the measurement. This means that if you clamp the leaf and then hit measure, the measurement will seem to take 45 seconds. Also, if you are hitting measure first, then clamping the leaf, and it is still taking 45 seconds to complete a measurement, take care to make sure that the clamp is closing completely.
Hopefully this clears up some confusion about best measurement practices and will help you get out there and up your data production. We here at PhotosynQ are cooking up bigger and better things for the next round of firmware and protocol updates. We are hoping we can mimic the ambient light through the PAR sensor and shine that from the device before we clamp so the leaf will never begin to dark adapt, pretty neat! Stick around for these updates.
Focusing on how the community is using PhotosynQ technologies. This month we are highlighting Matthew Daniel, an Arborist from Australia and one of PhotosynQ’s most active users.
Matthew is the director ofTree Preservation Australia and CEO of Global Urban Forest Pty Ltd, a company dedicated to the relationship between soil and tree health and the science of urban forestry. He travels to many cities in Australia and abroad collecting data on tree and soil health and prescribing proper health care programs to ensure that cities have happy, healthy trees. These prescriptions include compost tea’s, deep root soil injections, and vascular stem injections of specially formulated microbial plant and soil health inoculants and organic stimulants. However, his job was limited by the lack of affordable tools and the ability to share data and results with others.
How Matthew Connected with PhotosynQ
Matthew Daniel was born and raised in Tasmania, where he first developed his interest in trees and the outdoors in general. We asked him what his first memories with trees were and he told us “When I was 5 years old my uncle found me 40 feet up a tree, freaked my mum out”, no doubt this guy was destined to climb trees for a living! He would eventually receive training and certifications for arboriculture, working near high voltage power lines and in confined spaces, become a partner in Tree Preservation Australia and eventually founded Global Urban Forest Pty Ltd.
From the moment he first heard about PhotosynQ he knew “it was exactly what I needed to understand the tree health response to soil health intervention.” He has now completely integrated the PhotosynQ platform into his workflow saying “I use PhotosynQ before, during and after all the trees I work with.” That is a lot of trees!
He successfully applied to be a PhotosynQ beta tester and created his main project tree health calculator – Beta/Experts Program – 2015-2017, which has over 8,000 data points. Matthew recently received the new MultispeQ v1.0 and is working hard to build collaborations with community groups across Australia.
Ultimately, Matthew wants to use PhotosynQ as “an international collaborative platform to be a major part in quantifying and mitigating climate change.” Matthew believes that a global arborist community that shares data on a common, open platform can do more than improve the health of individual urban trees. It can also help to mitigate one of the causative factors of the Urban Heat Island Effect, namely a lack of vegetation in cities. Lack of vegetation in cities decreases levels of both evapotranspiration and carbon dioxide removal. It has been observed over the last century that cities are significantly warmer than the rural areas that surround them. The UHI effect affects many areas of life, such as the weather, health and the environment. It willincrease the production of rain clouds and thunder over your city, make youmore prone to violence during heat waves, increase your electricity bill, and evenkill off fish in lakes and streams just outside your city if mitigation processes are not undertaken. By going around Australian cities and learning about the trees that are present, Matthew and a global arborist community can learn which trees are most effective at mitigating the UHI effect and make your city a more enjoyable environment.
He knows this will not be easy, but it’s the potential that is driving Matthew. The potential to build a global arborist community that can collect data for cities that they can then utilize to manage their urban tree population, the potential to reduce the urban heat island effect, and finally, the potential to mitigate the effects of climate change.
Thank you to Matthew Daniel for allowing us to write about him and taking the time to answer all our questions. Hope to see you back in the US soon!
The PhotosynQ project started in David Kramers lab at Michigan State University, but it is by far not the only project, that is devoted to gain a deeper understanding into how plants do photosynthesis and how it is regulated. See what else is developed in this lab to study plant photosynthesis, all with the ultimate […]
Now that the MultispeQ v1.0 instruments are released, we want to share some stories of how the community is using PhotosynQ technolgies. Today we want to focus on Isaac Osei-Bonsu from Ghana, currently a PhD student in the Kramer Lab at Michigan State University.
Isaac Osei-Bonsu has been a PhotosynQ user since the early beta days in 2015. Over that time, he has collected over 13,000 measurements and created 46 PhotosynQ projects. As we worked towards releasing the new instruments, Isaac was often tasked with testing out new MultispeQ prototypes, and some iterations did not work so well! At the end of the day, Isaac collected over 3,000 measurements with MultispeQ v1.0 prototypes and his feedback helped us to modify and improve the MultispeQ throughout the design and manufacturing process.
Isaac hails from the west African nation of Ghana, where he was a research scientist for the Council for Scientific and Industrial Research-Crops Research Institute (CSIR-CRI). In Isaac’s own words, “The company is a National Agricultural Research institution which focuses on research on different crop species with the aim to improve agriculture in Ghana.” He studied a wide variety of crops including cowpea, peanut, pepper, eggplants, citrus, mangoes, avocado, pear, papaya and watermelon.
Ghana, like many countries in sub-Saharan Africa, is struggling to achieve food security and develop a robust economy. This means that the government wants farmers to produce more high value crops, like cocoa, for international markets. However, this comes at the expense of land for growing food crops, which is why it is so important to improve the productivity and efficiency of important food crops.
What Isaac is doing now
Isaac arrived at MSU with the help of a Legume Scholars Award which he received from the Consultative Group on International Agricultural Research (CGIAR) in 2015. The Legume Scholars Program supports the graduate study of young scientists from developing countries so they can pursue research careers involving grain legumes (http://legumelab.msu.edu/training/legume_scholars).
Isaac’s is studying the photosynthetic response of grain legumes to abiotic stress in order to improve grain legume production. His interest with automatic plant phenotyping was piqued “by the simple yet powerful nature of the MultispeQ device, connected to the PhotosynQ platform, and its possible use for rapid phenotyping in the field.” Manual phenotyping can be extremely slow and not entirely accurate. The MultispeQ instrument and the PhotosynQ platform make it easy and quick. He now uses the MultispeQ in most of his experiments. He doesn’t just use PhotosynQ because it is easier, but also because it allows him to develop a deeper understanding of abiotic stress responses in grain legumes.
Thank you to Isaac for taking some time out of your busy days to answer all of our questions. Having Isaac around our lab is a pleasure and his input into the PhotosynQ platform and MultispeQ instruments have been invaluable. We hope our instruments can help him as much as he has helped us and wish him luck in his continued research.
Our most recent website update probably got lost in the excitement about the news that we had started shipping the new MultispeQ v1.0. We have continued to work on the tutorials and data analysis and are happy to announce another update to https://photosynq.org. There are several libraries out there to graph data in the browser and when we started the platform, we decided to use Flot, a great, performance library, which is unfortunately no longer actively developed. Implementing new methods of plotting would have required us to build certain features from scratch. Instead we are now using Plot.ly which opened up its libraries to the public. It not only provides the library, but also its own platform to create and manipulate plots, as well as libraries for popular languages like Python orR. We would like to give you a quick update on what has changed and improved.
New and Improved Tutorials
While some people really like using YouTube videos to learn new technologies, others don’t find them very helpful. So, we updated the tutorials (available in the learn more menu from the top of the page) to include text pages that include step by step directions, pictures and screen caps to get you started with PhotosynQ. Also, you can now download a Getting Started manual in pdf form if you need to refer to the tutorials while working offline! We kept the the video’s for those you like them, you can find them in their own videos tab on the tutorials page.
New Plot Options
Scatter plots are a great way of looking at data down to individual measurements. At the same time it can be hard to see trends in a big cloud of points. We hope these new plotting options will help analyzing your data.
The 2D contour plot allows you to do exactly that. With additional histograms they are a great way to visualize populations within a big data set.
In addition to the already existing histograms, you can now plot two parameters as a 2D histogram.
Box plots are available now. Like the bar charts, you can use a category as well.
New Parameter – Time of Day
We added the parameter Time of Day, showing the time, when the measurement was recorded as a number (e.g. 1:30 pm = 13.5). This will be helpful to plot time dependent trends, which could not be visualized using a regular timescale, which could be the case when measurements were recorded over multiple days.
New Data Selection and Plot Capturing
With the new ways to plot data comes the ability to select a set of data points in a scatter plot using the box or lasso selection. It can be used to generate a selection based Series, or the data can be saved with the measurement identifiers as well as the meta data including time and the project questions and answers. The little camera icon enables you to save the current graph as an image (png or jpeg), including the graphs’ legend.
Using Python and R
Analyzing big datasets using the data viewer inside the browser might be difficult. Or you might want to do your own statistical analysis or plot the data in a way that is not available. For this reason we made packages for Python and R to help you get the most out of your data.
On your user page, you can now see your Network. Your Network includes people who collaborate with you and have joined one or more of your Projects, as well as all Project Leads of projects you have joined.
Tags and Project Categories are now displayed on your user page.
Tags are now links and can be searched. Just click on a tab or type your tag into the search field (e.g. #beans).
What’s No Longer Available
Plots – We said goodbye to a few features, because there was no good way of adding it to the new plotting library or the available options didn’t turn out to be all that useful.
The options of plotting spline lines and area charts.
“Time (normalized)” is no longer available. It got replaced by the parameter “Time of Day”
Dashboard – To make the dashboard a little easier we decided to remove the following options.
The Panel with a pie chart of total submitted and flagged. The totals are added to the other data quality panels.
We hope that these new features will help you to analyze your collected data.