Fighting Fusarium wilt to beat the bananapocalypse

Dr. Sarah Schmidt (@BananarootsBlog), Researcher and Science Communicator at The Sainsbury Laboratory Science. Sarah got hooked on both banana research and science writing when she joined a banana Fusarium wilt field trip in Indonesia as a Fusarium expert. She began blogging at https://bananaroots.wordpress.com and just filmed her first science video. She speaks at public events like the Pint of Science and Norwich Science Festival.

 

Every morning I slice a banana onto my breakfast cereal.

And I am not alone.

Every person in the UK eats, on average, 100 bananas per year.

Bananas are rich in fiber, vitamins, and minerals like potassium and magnesium. Their high carbohydrate and potassium content makes them a favorite snack for professional sports players; the sugar provides energy and the potassium protects the players from muscle fatigue. Every year, around 5000 kg of bananas are consumed by tennis players at Wimbledon.

But bananas are not only delicious snacks and handy energy kicks. For around 100 million people in Sub-Saharan Africa, bananas are staple crops vital for food security. Staple crops are those foods that constitute the dominant portion of a standard diet and supply the major daily calorie intake. In the UK, the staple crop is wheat. We eat wheat-based products for breakfast (toast, cereals), lunch (sandwich), and dinner (pasta, pizza, beer).

In Uganda, bananas are staple crops. Every Ugandan eats 240 kg bananas per year. That is around 7–8 bananas per day. Ugandans do not only eat the sweet dessert banana that we know; in the East African countries such as Kenya, Burundi, Rwanda, and Uganda, the East African Highland banana, called Matooke, is the preferred banana for cooking. Highland bananas are large and starchy, and are harvested green. They can be cooked, fried, boiled, or even brewed into beer, so have very similar uses wheat in the UK.

In West Africa and many Middle and South American countries, another cooking banana, the plantain, is cooked and fried as a staple crop.

In terms of production, the sweet dessert banana we buy in supermarkets is still the most popular. This banana variety is called Cavendish and makes up 47% of the world’s banana production, followed by Highland bananas (24%) and plantains (17%). Last year, I visited Uganda and I managed to combine the top three banana cultivars in one dish: cooked and mashed Matooke, a fried plantain and a local sweet dessert banana!

 

Three types of banana in a single dish in Uganda.

Another important banana cultivar is the sweet dessert banana cultivar Gros Michel, which constitutes 12% of the global production. Gros Michel used to be the most popular banana cultivar worldwide until an epidemic of Fusarium wilt disease devastated the banana export plantations in the so-called “banana republics” in Middle America (Panama, Honduras, Guatemala, Costa Rica) in the 1950s.

Fusarium wilt disease is caused by the soil-borne fungus Fusarium oxysporum f. sp. cubense (FOC). The fungus infects the roots of the banana plants and grows up through the water-conducting, vascular system of the plant. Eventually, this blocks the water transport of the plant and the banana plants start wilting before they can set fruits.

Fusarium Wilt symptoms

Fusarium Wilt symptoms

The Fusarium wilt epidemic in Middle America marked the rise of the Cavendish, the only cultivar that could be grown on soils infested with FOC. The fact that they are also the highest yielding banana cultivar quickly made Cavendish the most popular banana variety, both for export and for local consumption.

Currently, Fusarium wilt is once again the biggest threat to worldwide banana production. In the 1990s, a new race of Fusarium wilt – called Tropical Race 4 (TR4) – occurred in Cavendish plantations in Indonesia and Malaysia. Since then, TR4 has spread to the neighboring countries (Taiwan, the Philippines, China, and Australia), but also to distant locations such as Pakistan, Oman, Jordan, and Mozambique.

Current presence of Fusarium wilt Tropical Race 4. Affected countries are colored in red.

In Mozambique, the losses incurred by TR4 amounted to USD 7.5 million within just two years. Other countries suffer even more; TR4 causes annual economic losses of around USD 14 million in Malaysia, USD 121 million in Indonesia, and in Taiwan the annual losses amount to a whopping USD 253 million.

TR4 is not only diminishing harvests. It also raises the price of production, because producers have to implement expensive preventative measures and treatments of affected plantations. These preventive measures and treatments are part of the discussion at The World Banana Forum (WBF). The WBF is a permanent platform for all stakeholders of the banana supply chain, and is housed by the United Nation’s Food and Agricultural Organization (FAO). In December 2013, the WBF created a special taskforce to deal with the threat posed by TR4.

Despite its massive impact on banana production, we know very little about the pathogen that is causing Fusarium wilt disease. We don’t know how it spreads, why the new TR4 is so aggressive, or how we can stop it.

Fusarium Wilt symptom

Fusarium Wilt symptoms in the discolored banana corm.

Breeding bananas is incredibly tedious, because edible cultivars are sterile and do not produce seeds. I am therefore exploring other ways to engineer resistance in banana against Fusarium wilt. As a scientist in the 2Blades group at The Sainsbury Laboratory, I am investigating how we can transfer resistance genes from other crop species into banana and, more recently, I have been investigating bacteria that are able to inhibit the growth and sporulation of F. oxysporum. These biologicals would be a fast and cost-effective way of preventing or even curing Fusarium wilt disease.

 

Twitter:           @BananarootsBlog

Email:              mailto:[email protected]

Website:          https://bananaroots.wordpress.com

A taste of CRISPR

Dr Craig CormickThis week’s blog was written by Dr Craig Cormick, the Creative Director of ThinkOutsideThe. He is one of Australia’s leading science communicators, with over 30 years’ experience working with agencies such as CSIRO, Questacon and Federal Government Departments.

So what do you think CRISPR cabbage might taste like? CRISPR-crispy? Altered in some way?

Participants at the recent Society for Experimental Biology/Global Plant Council New Breeding Technologies workshop in Gothenburg, Sweden, had a chance to find out, because in Sweden CRISPR-produced plants are not captured by the country’s GMO regulations and can be produced.

Professor Stefan Jansson, one of the workshop organizers, has grown the CRISPR cabbage (discussed in his blog for GPC!) and not only had it included on the menu of the workshop dinner, but also had samples for participants to take away. Some delegates were keen to pick up the samples while others were unsure how their own country’s regulatory rules would apply to them.

 

 

Regulatory issues

The uncertainty some delegates felt about the legality of taking a CRISPR cabbage sample home was a good demonstration of the diversity of regulations that apply – or may apply – to new breeding technologies, such as CRISPR and gene editing – and there was considerable discussion at the workshop on how European Union regulations and court rulings may play out, affecting both the development and export/import of plants and foods produced by the new technologies.

A lack of certainty has meant many researchers are unable to determine whether their work will need to be subjected to costly and time-consuming regulations or not.

The need for new breeding technologies was made clear at the workshop, which was attended by 70 people from 17 countries, with presentations on the need to double our current food production to feed the world in 2050 and reduce crop losses caused by problems such as viruses, which deplete crops by 10–15%.

The two-day workshop, held in early July, looked at a breadth of issues, including community attitudes, gene editing success stories, and tools and resources. But discussions kept coming back to regulation.

Outdated regulations

Regulations of gene technologies were largely developed 20 years ago or so, for different technologies than now exist, and as a result are not clear enough for researchers to determine whether different gene editing technologies they are working on may be governed by them or not.

The diversity of regulations is also going to be an issue, for some countries may allow different gene editing technologies, but others may not allow products developed using them to be imported.

That led to the group beginning to develop a statement that captured the feeling of the workshop, which, when complete, it is hoped will be adopted by relevant agencies around the world to develop their own particular positions on gene editing technologies. It would be a huge benefit to have a coherent and common line in an environment of mixed regulations in mixed jurisdictions.

CRISPR cabbage

And as to the initial question of what CRISPR cabbage tastes like – just like any cabbage you might buy at your local supermarket or farmers market, of course – since it is really no different.

 

Want to read more about CRISPR? Check out our interview with Prof. Stefan Jansson or our introduction to CRISPR from Dr Damiano Martignago.

Brazil’s transgenic sugarcane stirs up controversy

By Luisa Massarani

This article was originally published on SciDev.Net. Read the original article.

[RIO DE JANEIRO] A genetically modified (GM) cane variety that can kill the sugarcane borer (Diatraea saccharalis) has been approved in Brazil,  to the delight of some scientists and the dismay of others, who say it may threaten Brazilian biodiversity.

Brazil is the second country, after Indonesia, to approve the commercial cultivation of GM sugarcane. The approval was announced by the Brazilian National Biosafety Technical Commission (CTNBio) on June 8.

Sugarcane borer is one of the main pests of the sugarcane fields of South-Central Brazil, causing losses of approximately US$1.5 billion per year.

“Breeding programmes could not produce plants resistant to this pest, and the existing chemical controls are both not effective and severely damaging to the environment,” says Adriana Hemerly, a professor at the Federal University of Rio de Janeiro, in an interview with SciDev.Net.

“Studies conducted outside Brazil prove that protein from genetically modified organisms harms non-target insects, soil fauna and microorganisms.”

Rogério Magalhães

“Therefore, the [GM variety] is a biotechnological tool that helps solve a problem that other technologies could not, and its commercial application will certainly have a positive impact on the productivity of sugarcane in the country.”

Jesus Aparecido Ferro, a member of CTNBio and professor at the Paulista Júlio de Mesquita Filho State University, believes the move followed a thorough debate that began in December 2015 — that was when the Canavieira Technology Center (Sugarcane Research Center) asked for approval to commercially cultivate the GM sugarcane variety.

“The data does not provide evidence that the cane variety has a potential to harm the environment or human or animal health,” Ferro told SciDev.Net.

To develop the variety, scientists inserted the gene for a toxin [Cry] from the bacterium Bacillus thuringiensis (Bt) into the sugarcane genome, so it could produce its own insecticide against some insects’ larvae.

This is a technology that “has been in use for 20 years and is very safe”, says Aníbal Eugênio Vercesi, another member of the CTNBio, and a professor at the State University of Campinas.

But Valério De Patta Pillar, also a member of the CTNBio and a professor at the Federal University of Rio Grande do Sul, points to deficiencies in environmental risk assessment studies for the GM variety — and the absence of assessments of how consuming it might affect humans and animals.

According to Pillar, there is a lack of data about the frequency with which it breeds with wild varieties. Data is also missing on issues such as the techniques used to create the GM variety and the effects of its widespread use.

Rogério Magalhães, an environmental analyst at Brazil’s Ministry of the Environment, also expressed concern about the approval of the commercial transgenic cane.

“I understand that studies related to the impacts that genetically modified sugarcane might have on Brazilian biodiversity were not done by the company that owns the technology,” said Magalhães in an interview with SciDev.Net. This is very important because Brazil’s climate, species, and soils differ from locations where studies might have taken place, he explained.

Among the risks that Magalhães identified is contamination of the GM variety’s wild relatives. “The wild relative, when contaminated with transgenic sugarcane, will have a competitive advantage over other uncontaminated individuals, as it will exhibit resistance to insect-plague that others will not have,” he explained.

Another risk that Magalhães warns about is damage to biodiversity. “Studies conducted outside Brazil prove that Cry protein from genetically modified organisms harms non-target insects, soil fauna and microorganisms.”

Magalhães added that some pests have already developed resistance to the Bt Cry protein, prompting farmers to apply agrochemicals that are harmful to the environment and human health.

This piece was originally published by SciDev.Net’s Latin America and Caribbean desk.

 

This article was originally published on SciDev.Net. Read the original article.

Striga hermonthica – a beautiful but devastating plant…

This week’s post was written by Caroline Wood, a PhD candidate at the University of Sheffield.

When it comes to crop diseases, insects, viruses, and fungi may get the media limelight but in certain regions it is actually other plants which are a farmer’s greatest enemy. In sub-Saharan Africa, one weed in particular – Striga hermonthica – is an almost unstoppable scourge and one of the main limiting factors for food security.

Striga is a parasitic plant; it attaches to and feeds off a host plant. For most of us, parasitic plants are simply harmless curiosities. Over 4,000 plants are known to have adopted a parasitic mode of life, including the seasonal favorite mistletoe (a stem parasite of conifers) and Rafflesia arnoldii, nicknamed the “corpse flower” for its huge, smelly blooms. Although the latter produces the world’s largest flower, it has no true roots – only thread-like structures that infect tropical vines.

When parasitic plants infect food crops, they can turn very nasty indeed. Striga hermonthica is particularly notorious because it infects almost every cereal crop, including rice, maize, and sorghum. Striga is a hemiparasite, meaning that it mainly withdraws water from the host (parasitic plants can also be holoparasites, which withdraw both water and carbon sugars from the host). However, Striga also causes a severe stunting effect on the host crop (see Figure 1), reducing their  yield to practically nothing. Little wonder then, that the common name for Striga is ‘witchweed’.

Striga-infected sorghum

Figure 1: Striga-infected sorghum. Note the withered, shrunken appearance of the infected plants. Image credit: Joel Ransom.

 

Several features of the Striga lifecycle make it especially difficult to control. The seeds can remain dormant for decades and only germinate in response to signals produced by the host root (called strigolactones) (Figure 2). Once farmland becomes infested with Striga seed, it becomes virtually useless for crop production. Germination and attachment takes place underground, so the farmer can’t tell if the land is infected until the parasite sends up shoots (with ironically beautiful purple flowers). Some chemical treatments can be effective but these remain too expensive for the subsistence farmers who are mostly affected by the weed. Many resort to simply pulling the shoots out as they appear; a time-consuming and labor-intensive process. It is estimated that Striga spp. cause crop losses of around US $10 billion each year [1].

Certain crop cultivars and their wild relatives show natural resistance to Striga. Here at the University of Sheffield, our lab group (headed by Professor Julie Scholes) is working to identify resistance genes in rice and maize, with the eventual aim of breeding these into high-yielding cultivars. To do this, we grow the host plants in rhizotrons (root observation chambers) which allow us to observe the process of Striga attachment and infection (see Figure 3). Already this has been successful in identifying rice cultivars that have broad-spectrum resistance to Striga, and which are now being used by farmers across Africa.

 

Life cycle of Striga

Figure 2: Life cycle of Striga spp. A single plant produces up to 100,000 seeds, which can remain viable in the soil for 20 years. Following a warm, moist conditioning phase, parasite seeds become responsive to chemical cues produced by the roots of suitable hosts, which cause them to germinate and attach to the host root. The parasite then develops a haustorium: an absorptive organ which penetrates the root and connects to the xylem vessels in the host’s vascular system. This fuels the development of the Striga shoots, which eventually emerge above ground and flower. Figure from [2].

 

But many fundamental aspects of the infection process remain almost a complete mystery, particularly how the parasite overcomes the host’s intrinsic defense systems. It is possible that Striga deliberately triggers certain host signaling pathways; a strategy used by other root pathogens such as the fungus Fusarium oxysporum. This is the focus of my project: to identify the key defense pathways that determine the level of host resistance to Striga. It would be very difficult to investigate this in crop plants, which typically have incredibly large genomes, so my model organism is Arabidopsis thaliana, the workhorse of the plant science world, whose genome has been fully sequenced and mapped. Arabidopsis cannot be infected by Striga hermonthica but it is susceptible to the related species, Striga gesnerioides, which normally infects cowpea.  I am currently working through a range of different Arabidopsis mutants, each affected in a certain defense pathway, to test whether these have an altered resistance to the parasite.  Once I have an idea of which plant defense hormones may be involved (such as salicylic acid or jasmonic acid), I plant to test the expression of candidate genes to decipher what is happening at the molecular level.

Striga-infected Arabidopsis

Figure 3: One of my Arabidopsis plants growing in a rhizotron. Preconditioned Striga seeds were applied to the roots three weeks ago with a paintbrush. Those that successfully attached and infected the host have now developed into haustoria. The number of haustoria indicates the level of resistance in the host. Image credit: Caroline Wood.

 

It’s early days yet, but I am excited by the prospect of shedding light on how these devastating weeds are so effective in breaking into their hosts. Ultimately this could lead to new ways of ‘priming’ host plants so that they are armed and ready when Striga attacks. It’s an ambitious challenge, and one that will certainly keep me going for the remaining two years of my PhD!

 

You can follow my journey by reading my blog and keeping up with me on Twitter (@sciencedestiny).

 

References:

[1] Westwood, J. H. et al. (2010). The evolution of parasitism in plants. Trends in Plant Science, 15(4): 227-235.

[2] Scholes, J. D. and Press, M. C. (2008). Striga infestation of cereal crops – an unsolved problem in resource limited agriculture. Current Opinion in Plant Biology, 11(2): 180-186.

Just add water: Could resurrection plants help feed the world?

This week we spoke to Professor Henk Hilhorst (Wageningen University and Research) about his research on desiccation tolerance in seeds and plants.

 

Could you begin by telling us a little about your research?

I am a plant physiologist specializing in seed biology. I have a long research record on various aspects of seeds, including the mechanisms and regulation of germination and dormancy, desiccation tolerance, as well as issues in seed technology. Being six years from retirement now, I decided to extend my desiccation tolerance studies from seeds to resurrection plants, which display vegetative desiccation tolerance. I strongly believe that unveiling of the mechanism of vegetative desiccation tolerance may help us create crops that are truly tolerant to severe drought, rather than (temporarily) resistant.

 

How did you become interested in this field of study, and how has your career progressed?

As with many things in life, it was coincidence. I majored in plant biochemistry and applied for a PhD position in seed biology. After obtaining the degree I was offered a tenure track position in seed physiology by the Laboratory of Plant Physiology at Wageningen University, where I still work as a faculty member. My career has progressed nicely and I am an authority in the field of seed science, editor-in-chief of the journal Seed Science Research, and will become the President of the International Society for Seed Science in September of this year.

I see my current work on vegetative desiccation tolerance as a highlight in my professional life. I have always been more interested in the desiccation tolerance of seeds until about five years ago, when my current collaborator Prof Jill Farrant of the University of Cape Town, South-Africa, made me enthusiastic about these wonderful resurrection plants. We started to work together and published our first study recently in Nature Plants.

Read the paper here ($): A footprint of desiccation tolerance in the genome of Xerophyta viscosa.


 

In your recent paper, you sequenced the genome of the resurrection plant, Xerophyta viscosa, which can survive with less than a 5% relative water content. How is it possible for a plant to lose so much of its water and still survive?

These plants have a lot of characteristics that we’ve seen in seeds. They display protective desiccation tolerance mechanisms in their leaves, including anti-oxidants, protective proteins, and even dismantle their photosynthetic machinery during periods of drought. Even the cell wall structure and composition of resurrection plants resemble those of seeds. We are currently working on a paper describing the striking similarities between seeds and resurrection plants.

 

What was the most interesting discovery you made upon sequencing the genome of the resurrection plant?

First, the similarities between resurrection plants and seeds listed above were also apparent at the molecular level. For example, previous work suggested that the “ABI3 regulon”, consisting of about 100 genes regulated by the transcription factor ABI3, is specific to seeds, but we found that it is almost completely present (and active) in the leaves of Xerophyta viscosa too!

Secondly, we found “islands” or clusters of genes specific for desiccation tolerance that aren’t found in other species. Many of these regulate secondary metabolite pathways.

 

How challenging was it to sequence the genome of this plant? How did you overcome any difficulties?

It was very challenging. First, the species is an octoploid, meaning it has eight copies of each chromosome. This meant that we had to sequence its genome at very high coverage and employing the most advanced sequencing facilities, e.g. PacBio. Getting funding for this complex analysis was another challenge. We then took almost a year to assemble the genome and annotate it at the desired quality.

 

Xerophyta viscosa

Xerophyta viscosa before and after the rains. Image credit: Prof. Henk Hilhorst.

 

You identified some of the most important genes involved in desiccation tolerance. Is it possible to translate this work into other species, such as crops that may be threatened by drought as the climate changes?

That will be our ultimate goal. It’s important to remember that desiccation-sensitive plants, including all our major crops, produce seeds that are desiccation tolerant. This implies that the information for desiccation tolerance is present in the genomes of these crops but that it is only turned on in the seeds. We are trying to determine how this is localized, in order to find a method to turn on the desiccation tolerance mechanism in vegetative parts of the (crop) plant too. In parallel we are expressing some of the key transcription factors from Xerophyta viscosa in some important crops to see how this affects them.

 

Are there any other interesting aspects of Xerophyta viscosa biology?

Contrary to plants that wilt and ultimately die because of (severe) drought, leaves of resurrection species do not show such stress-related senescence. This is related to the engagement of active anti-senescence genes during the drying of the leaves of resurrection species. We are currently investigating these senescence-related mechanisms too.

 

Rose of Jericho (Anastatica hierochuntica)

The rose of Jericho (Anastatica hierochuntica) is another resurrection plant. Image credit: FloraTrek. Used under license: CC BY-SA 3.0.

 

Do you expect to find that different types of desiccation-tolerant plants use the same subset of genes to survive drought, or could they have developed other pathways to resilience?

We expect that the core mechanism is very similar among the resurrection species but that each species may have adapted to its specific environment.

Funding permitting, we will sequence the genomes of at least another ten resurrection species to further clarify the various evolutionary pathways to desiccation tolerance and, importantly, to discriminate between species-specific and desiccation tolerance-specific genes.

 

What advice do you have for early career researchers?

Stick to what you believe in, even if you have to (temporarily) be involved in research that you appreciate less, e.g., because of better funding opportunities.

 


Read Henk’s recent paper in Nature Plants here ($): A footprint of desiccation tolerance in the genome of Xerophyta viscosa.

Roots of a second green revolution

This week we spoke to Professor Jonathan Lynch, Penn State University, whose research on root traits has deepened our understanding of how plants adapt to drought and low soil fertility.

 

 

Could you begin by giving us a brief introduction to your research?

We are trying to understand how plants adapt to drought and low soil fertility. This is important because all plants in terrestrial ecosystems experience suboptimal water and nutrient availability, so in rich nations we maintain crop yields with irrigation and fertilizer, which is not sustainable in the long term. Furthermore, climate change is further degrading soil fertility and increasing plant stress. This topic is therefore both a central question in plant evolution and a key challenge for our civilization. We need to develop better ways to sustain so many people on this planet, and a big part of that will be developing more resilient, efficient crop plants.

 

Drought is devastating for crops

Drought and low soil fertility are devastating for crops. Image credit: CIAT. Used under license: CC BY-SA 2.0.

 

What got you interested in this field, and how has your career developed over time?

When I was 9 years old I became aware of a famine in Africa related to crop failure and resolved to do something about it. I studied soils and plant nutrition as an undergraduate, and in graduate school worked on plant adaptation to low phosphorus and salinity stress, moving to a research position at the CIAT headquarters in Colombia. Later I moved to Penn State, where I have maintained this focus, working to understand the stress tolerance of staple crops, and collaborating with crop breeders in the USA, Europe, Africa, Asia, and Latin America.

 

Your recent publications feature a variety of different crop plants. Could you talk about how you select a species to study?

We work with species that are important for food security, that grow in our field environments, and that I think are cool. We have devoted most of our efforts to the common bean – globally the most important food legume – and maize, which is the most important global crop. These species are often grown together in Africa and Latin America, and part of our work has been geared to understanding how maize/bean and maize/bean/squash polycultures perform under stress. These are fascinating, beautiful plants with huge cultural importance in human history. They are also supported by talented, cooperative research communities. One nice feature of working with food security crops is that their research communities share common goals of achieving impact to improve human welfare.

 

Common bean (Phaseolus vulgaris)

The common bean (Phaseolus vulgaris) is an important staple in many parts of the world. Image credit: Ervins Strauhmanis. Used under license: CC BY 2.0.

 

Many researchers use Arabidopsis thaliana for plant research, but are crops better suited for root research than the delicate roots of Arabidopsis? Are crop plants more or less difficult to work with in your research than Arabidopsis?

The best research system is entirely a function of your goals and questions. We have worked with Arabidopsis for some questions. Since we work with processes at multiple scales, including crop stands, whole organisms, organs, tissues, and cells, it has been useful to work with large plants such as maize, which are large enough to easily measure and to work with in the field. The most interesting stress adaptations for crop breeding are those that differ among genotypes of the same species, and at that level of organization there is a lot of biology that is specific to that species, that cannot readily be generalized from model organisms with very different life strategies. There has been considerable attention to model genomes and much less attention to model phenomes.

 

You have developed methodologies for the high-throughput phenotyping of crop plants. What does this technique involve and what challenges did you have to overcome to succeed?

We have developed multiple phenotyping approaches – too many to summarize readily here. Our overall approach is simply to develop a tool that helps us achieve our goals. For example, we have developed tools to quantify the root architecture of thousands of plants in the field, to measure anatomical phenotypes of thousands of samples from field-grown roots, to help us determine which root phenotypes might affect soil resource capture, etc. Working with geneticists and breeders, we are constantly asked to measure something meaningful on thousands of plants in a field, in many fields, every season. ARPA-E (the US Advanced Research Projects Agency for Energy) has recently funded us to develop phenotyping tools for root depth in the field, but this is the first time we have been funded to develop phenotyping tools – generally we just come up with things to help us do our work, which fortunately have been useful for other researchers as well.

 

Could you talk about some of the computational models you have developed for investigating plant growth and development?

The biological interactions between plants and their environment are so complex, we need computational (in silico) tools to help us evaluate them. Increasingly, in silico tools can integrate information across multiple scales, from gene expression to crop stands. These tools also allow us to evaluate things that are difficult to measure, such as phenotypes that do not yet exist, or future climates. In silico biology will be an essential tool in 21st Century biology, which will have access to huge amounts of data at multiple scales that can be used to try to understand incredibly complex systems, such as the human brain or roots interacting with living soil. Our main in silico tool is SimRoot, developed over the past 25 years to understand how root phenotypes affect soil resource capture.

Check out a SimRoot model below:


 

You have been working on breeding plants that have improved yield in soils with low fertility. What have you achieved in this work?

In collaboration with crop breeders and colleagues in various nations we have developed improved common bean lines with better yield under drought and low soil fertility that are being deployed in Africa and Latin America, improved soybean lines with better yield in soils with low phosphorus being deployed in Africa and Asia, and are now working with maize breeders in Africa to develop lines with better yield under drought and low nitrogen stress. Many crop breeders are using our methods for root phenotyping to target root phenotypes in their selection regimes in multiple crops.

 

What piece of advice do you have for early career researchers?

You are at the forefront of an unprecedented challenge we face as a species – how to sustain 10 billion people in a degrading environment. Plant biologists are an essential part of the effort to reshape how we live on this planet. Do not doubt the importance of your efforts. Do not lose sight of the very real human impact of your scientific choices. Do not be deterred by the gamesmanship and ‘primate politics’ of science. You can make a difference. We need you.

Synthetic biology in chloroplasts

Dr Anil Day, University of Manchester

Dr Anil Day, University of Manchester

This week we spoke to Dr. Anil Day, a synthetic biologist at the University of Manchester who has developed an impressive array of tools and techniques to transform chloroplast genomes.

 

Could you begin by giving our readers a brief overview of synthetic biology?

Synthetic biology involves the application of engineering principles to biological systems. One approach to understanding a biological system is to break it down into smaller parts, which can be used to design new properties. These redesigned pieces can be reassembled into a new system, tested experimentally, and refined in an iterative process. Synthetic biology projects that are underway in our lab include designing plastids such as chloroplasts with new metabolic functions, and in the longer term the design and assembly of synthetic chloroplast genomes.

 

Anil Day examines transformed plants

Dr. Anil Day examines a cabinet of transformed plants. Credit: Dr. Anil Day.

Why do you use chloroplasts for synthetic biology systems?

Chloroplasts have a relatively small genome, coding for about 100 genes. Importantly, exogenous (foreign) genes coding for new functions can be precisely introduced into the chloroplast genome. All of the plastids within a plant contain the same genome so, once established, the user-designed reprogrammed plastids will be present throughout the plant. Chloroplasts can also produce very high levels of protein; researchers have achieved expression levels where over 70% of the total soluble protein in the leaves is the engineered protein. Expression in tomato fruit is also possible.

Multiple genes can be introduced into chloroplasts and expressed coordinately, allowing the metabolic engineering of more complex processes. The upper size limit for insertions is not known but is likely to be above the 50,000 nucleotide insertion achieved to date. Furthermore, chloroplasts and other plastids are important metabolic hubs and contain a wide variety of chemical substrates useful for metabolic engineering.

Plastids in plants

Plants have several types of plastids, including green photosynthetic chloroplasts, pigment-containing chromoplasts, and starch-containing amyloplasts. Credit: Dr. Anil Day.

 

Could you describe the current state of our ability to engineer chloroplasts?

Chloroplast engineering is routine in many labs around the globe. Although there are multiple chloroplasts in every cell, the process of converting all the chloroplasts to a single population of engineered genomes is not an issue. Most researchers use the tobacco plant because it is easily transformed, but other crops are amenable to transformation, including oilseed rape, soybean, tomato, and potato (cereals such as rice and wheat are more problematic). There has been progress with developing the inducible expression of exogenous genes in chloroplasts too.

 

What challenges/differences do you face when transforming chloroplast genomes when compared to the nuclear genome?

Typical genetic modification of the DNA in the nucleus is performed by introducing exogenous genes in T-DNA. T-DNA is transferred to the plant using the bacterium Agrobacterium tumefaciens, which is an efficient process, but the T-DNA integrates ‘randomly’ at many sites within chromosomes and different lines can have variable expression levels due to positional effects and gene silencing.

A. tumefaciens-mediated gene delivery systems do not work for chloroplast transformation. Most chloroplast transformation labs introduce genes into plastids by blasting cells with gold or tungsten particles coated with DNA. Because chloroplast genomes are present in multiple copies per cell, the process of converting all resident chloroplasts to the transgenic genome requires a continued period of selection. This means that the isolation of chloroplast transformants can take slightly longer than nuclear transformation. In our lab, we speed up this process by using restoration of photosynthesis to select chloroplasts with exogenous genes. Once plants with a uniform population of transgenic plastid genomes have been isolated, the transgenes are stable and inherited through the maternal line.

For the novice, I would say nuclear transformation using A. tumefaciens is easier to accomplish than chloroplast transformation.

 

Edited chloroplasts

A tobacco plant containing leaf areas with edited (pale green) and normal (darker green) chloroplasts. Credit: Dr. Anil Day.

Last year you reported that chloroplasts degrade in mature sperm cells just prior to fertilization. Could you elaborate on how this might be utilized in future crop breeding?

Chloroplasts are inherited from the female parent in wheat. This is useful because it restricts the pollen-mediated spread of chloroplast-localized transgenes into the environment. Previously, no-one had studied the mechanism of maternal chloroplast inheritance in wheat using modern cell biology tools. With our collaborators Lucia PrimavesiHuixia Wu, and Huw Jones at Rothamsted Research, we developed an efficient method to observe small non-green plastids in wheat pollen in real time. We found that the plastids were destroyed during the maturation of sperm cells, which explained the absence of paternal plastids in the offspring.

This discovery has applications in crop breeding. Anther culture is a powerful technique where new homozygous plants can be produced by doubling the chromosome numbers of haploid plants regenerated from pollen. This technique has been challenging in cereals, as chloroplast degradation in pollen leads to a high percentage of albino plants (in some cases 100% albinos). Understanding how to prevent the destruction of plastids in pollen sperm cells will improve this technique in cereals, which could speed up crop breeding in the future.

 

Selection of transformed plants

Transformed plantlets are selected by their ability to survive on a herbicide-containing agar plate, and can then be grown up into mature plants. Credit: Dr. Anil Day.

 What sorts of processes have you successfully transformed into chloroplasts, and what kinds of results have you achieved?

We have expressed a variety of exogenous genes in chloroplasts, from those conferring resistance to herbicides to vaccine epitopes and pharmaceutical proteins:

  • Plants expressing the bar gene in chloroplasts were resistant to the herbicide glufosinate (also known as phosphinothricin).
  • A chloroplast-expressed viral epitope was used to identify samples of human blood infected with the hepatitis C virus.
  • Human transforming growth factor 3 (hTGFβ3), a potential wound healing drug, accumulated to high concentrations in chloroplasts, and could be processed to a pure active form resembling clinical grade hTGFβ3.
  • In collaboration with Ray Dixon, Cheng Qi, and Mandy Dowson-Day at the John Innes Centre, we investigated the feasibility of introducing nitrogen-fixing genes into chloroplasts. This work was initiated in a unicellular green alga with the bacterial nifH gene.

 

What is the cutting edge of chloroplast transformation research?

Chloroplast genes are important for plant growth and development but they are difficult to improve by conventional breeding methods. We recently developed a method to edit plastid genomes, which allows beneficial single point mutations to be introduced into chloroplast genes. This is important because the resulting plants have an identical genome to the original cultivar apart the single base substitution, potentially leading to a new class of biotech crop.

Lentils under the lens: Improving genetic diversity for sustainable food security

This week’s post comes to us from Crystal Chan, project manager of the Application of Genomic Innovation in the Lentil Economy project led by Dr. Kirstin Bett at the Department of Plant Sciences, University of Saskatchewan.

 

Could you begin with a brief introduction to your research?

Our research focuses on the smart use of diverse genetic materials and wild relatives in the lentil (Lens culinaris) breeding program.

Canada has become the world’s largest producer and exporter of lentils in recent years. Lentils are an introduced species to the northern hemisphere and, until recently, our breeding program at the University of Saskatchewan involved just a handful of germplasms adapted to our climatic condition. With dedicated breeding efforts we have achieved noteworthy genetic gains in the past decade, but we are missing out on the vast genetic diversity available within the Lens genus. This is a major dilemma faced by all plant breeders: do we want consistency (sacrificing genetic diversity and reducing genetic gains over time) or diversity (sacrificing some important fixed traits and spending lots of time and resources in “backcrossing/rescue efforts”)?

 

In our current research, we use genomic tools to understand the genetic variability found in different lentil genotypes and the basis of what makes lentils grow well in different global environments (North America vs. Mediterranean countries vs. South Asian countries). We will then develop molecular breeding tools that breeders can use to improve the diversity and productivity of Canadian lentils while maintaining their adaptation to the northern temperate climate.

 

What first led you to this research topic?

Dr. Albert (Bert) Vandenberg, professor and lentil breeder at the University of Saskatchewan, noticed one of the wild lentil species was resistant to several diseases that devastate the cultivated lentil. After years of dedicated breeding effort, he was able to transfer the resistance traits to the cultivated lentil, but it took a lot of time and resources. We began looking into other beneficial traits and became fascinated with the domestication and adaptation aspects of lentil – after all lentil is one of the oldest cultivated crops, domesticated by man around 11,000 BC! With the rapid advance in genomic technology, we can start to better understand the biology and develop tools to harness these valuable genetic resources.

 

You have been involved in the development of tools that assist researchers to build databases of genomics and genetics data. Could you tell us more about projects such as Tripal?

Over the past six years, Lacey Sanderson (bioinformaticist in our group) has developed a database for our pulse research program at the University (Knowpulse, http://knowpulse.usask.ca/portal/). The database is specifically designed to present data that is relevant to breeders, as our group has a strong focus on variety development for the Canadian pulse crop industry. Knowpulse houses genotypic information from past and on-going lentil genomics projects, and includes tools for looking up genotypes as well as comparing the current genome assembly (currently v1.2) and other sequenced legume genomes. The tools are being developed in Tripal, an open-source toolkit that provides an interface between the data and a Drupal web content management system, in collaboration with colleagues at Washington State University.

 

At the moment we are developing new functionalities that will allow us to store and present germplasm information as well as phenotypic data. We are also working with our colleagues at Washington State University (under the “Tripal Gateway Project” funded by the National Science Foundation) to enhance interconnectivity between Knowpulse and other legume databases, such as the Legume Information Service (LIS) and Soybase, to facilitate comparative genomic studies.

How challenging are pulse genomes to assemble? How closely related are the various crops?

We had the fortune to lead the lentil genome sequencing initiative thanks to the support from producer groups and governments across the globe.  The lentil genome is really challenging to assemble! We see nice synteny between lentil and the model legume, medicago, however the lentil genome is much bigger. We see a significant increase in genome size between chickpea and beans versus lentil (and pea for that matter), yet we have evidence to show that genome duplication is not the cause of the size increase. There are a lot of very long repetitive elements sprinkled around the genome, which makes its sequencing and proper assembly very challenging. Not to mention understanding the role of these long repetitive elements in biological functions…

 

What insights into crop domestication have you gained from these genomes?

That’s what we are working on right now under the AGILE (“Application of Genomics to Innovation in the Lentil Economy”) project. Stay tuned!

 

Do you work with breeders to develop new cultivars? What sorts of traits are most important? 

Breeding is at the core of our work – both Kirstin and Bert are breeders (Kirstin has an active dry bean breeding program when she’s not busy with genomic research). All our research aims to feed information to the breeders so that they can make better crossing and selection decisions. Our work in herbicide tolerance has led to the development and implementation of a molecular marker to screen for herbicide resistance. With that marker we save time (skipping a crossing cycle) and forego the herbicide spraying test for all of our early materials.

Disease resistance and drought tolerance are also important for the growers. Visual quality (seed shape, size, color) are very important too as our customers are very picky as to what sort of lentils they like to buy/eat.

What does the future of legume/lentil agriculture hold?

Lentils have been a staple food in many countries for centuries and have been gaining popularity in North America in recent years as people are looking for plant-based protein sources. Lentils are high in fibre, protein, and complex carbohydrates, while low in fat and calories, and have a low glycemic index. They are suitable for vegetarian/vegan, gluten-free, diabetic, and heart-smart diets. Lentils also provide essential micronutrients such as iron, zinc and folates. Lentils are widely recognized as nutrient-dense food that could serve as part of the solution to combat global food and nutritional insecurity.

In modern agriculture, adding lentil or other leguminous crops in the crop rotation helps improve soil structure, soil quality, and biotic diversity, as well as enhancing soil fertility through their ability to fix nitrogen. Because pulse crops require little to no nitrogen fertilizer, they use half of the non-renewable energy inputs of other crops, reducing greenhouse gas emissions.

2016 was marked by the United Nations as the International Year of Pulses, which was great as many people have become more aware of the benefits of pulse crops on the plate and in the field.

 

Follow us on twitter (@Wildlentils) for research updates!

 

All images are credited to Mr Derek Wright.

Mother grain genome: insights into quinoa

Sales of quinoa (Chenopodium quinoa) have exploded in the last decade, with prices more than tripling between 2008 and 2014. The popularity of this pseudocereal comes from its highly nutritious seeds, which resemble grains and contain a good balance of protein, vitamins, and minerals. The nourishing nature of quinoa meant it was prized by the Incas, who called it the “Mother grain”.

Quinoa

Quinoa is a popular ‘grain’, but it is more closely related to spinach and beetroot than cereals like wheat or barley. Image credit: Flickr user. Used under license: CC BY 2.0.

Quinoa is native to the Andes of South America, where it thrives in a range of conditions from coastal regions to alpine regions of up to 4000 m above sea level. Its resilience and nutritious seeds means that quinoa has been identified as a key crop for enhancing food security, but there are currently very few breeding programs targeting this species.

The challenge of improving the efficiency and sustainability of quinoa production has so far been restricted by the lack of a reference genome. This week, a team of researchers led by Professor Mark Tester (King Abdullah University of Science & Technology; KAUST) addressed this issue, publishing a high-quality genome sequence for quinoa in Nature. They compared the genome with that of related species to characterize the evolution and domestication of the crop, and investigated the genetic diversity of economically important traits.

 

The evolution of quinoa

Tester and colleagues used an array of genomics techniques to assemble 1.39 Gb of the estimated 1.45-1.50 Gb full length of quinoa’s genome. Quinoa is a tetraploid, meaning it has four copies of each chromosome. The researchers shed light on the evolutionary history of this crop by sequencing descendants of the two diploid species (each containing two sets of chromosomes) that hybridized to generate quinoa; kañiwa (Chenopodium pallidicaule) and Swedish goosefoot (Chenopodium suecicum). Comparing these sequences to quinoa and other relatives, the team showed that the hybridization event likely occurred between 3.3 and 6.3 million years ago. A comparison with other closely related Chenopodium species also suggested that, contrary to previous predictions, quinoa may have been domesticated twice, both in highland and coastal environments.

Quinoa field

Quinoa field. Image credit: LID. Used under license: CC BY-SA 2.0.

 

Washing away quinoa’s bitter taste

Quinoa seeds are coated with soap-like chemicals called saponins, which have a bitter taste that deters herbivores. Saponins can disrupt the cell membranes of red blood cells, so they have to be removed before human consumption, but this process is costly, so quinoa breeders are always looking for varieties that produce lower levels of saponins.

Sweet (low-saponin) quinoa strains do occur naturally, but the genes that regulate this phenotype were previously unknown. Tester and colleagues investigated sweet and bitter quinoa strains and discovered that a single gene (TRITERPENE SAPONIN BIOSYNTHESIS ACTIVATING REGULATOR-LIKE 1 [TSARL1]) controls the amount of saponins produced in the seeds. The low-saponin quinoa strains contained mutations in TSARL1 that prevented it from functioning properly. This is a key target for the improvement of quinoa in the future, although farmers will have to find new ways to protect their crops from birds and other seed predators!

Quinoa flowers

Quinoa flowers. Image credit: Alan Cann. Used under license: CC BY-SA 2.0.

 

Quality quinoa

The high-quality reference genome for quinoa generated by Tester and colleagues is likely to be vital for allowing many exciting improvements in the future. Breeders hoping to improve the yield, ease of harvest, stress tolerance, and saponin content of quinoa can develop genetic markers to speed up breeding for these key traits, improving the productivity of quinoa varieties and enhancing future food security.

 


Read the paper in Nature: Jarvis et al., 2017. The genome of Chenopodium quinoa. Nature. DOI: 10.1038/nature21370

Thank you to Professor Mark Tester (KAUST) for providing information used in this post!

The future of phenotyping

This week’s post was written by Dr Kasra Sabermanesh, Rothamsted Research.

I am a post-doctoral research scientist within Rothamsted Research’s BBSRC-funded 20:20 Wheat® program, which aims to provide the knowledge base and tools to increase the UK wheat yield potential from 8.4 to 20 tons of wheat per hectare within the next 20 years. Field phenotyping is one component of this program and facilitates the non-destructive monitoring of field-grown crops. Traditional methods of field phenotyping require huge human effort, which consequently limits the accuracy, frequency, and number of different measurements that can be taken at one time. Fortunately, Rothamsted has an exciting solution to this problem.

The Field Scanalyzer

Kasra Sabermanesh

Dr Kasra Sabermanesh shows off the Field Scanalyzer. Image credit: Rothamsted Research

Our field phenotyping platform, the Field Scanalyzer (constructed by LemnaTec GmbH and being further developed by ourselves), supports a motorized measuring platform with multiple sensors that can be accurately positioned anywhere within a dedicated field. The sensor array comprises a high-definition RGB camera, two hyperspectral cameras, a thermal infrared camera, a system for imaging chlorophyll fluorescence and twin scanning lasers for 3D information capture. Together, these sensors generate a wealth of data about crop growth, architecture, performance, and health. The Field Scanalyzer operates autonomously and in high-throughput, meaning it can take a lot of non-destructive measurements without human supervision, throughout the crops lifecycle, with high-accuracy and reproducibility. (You can read more about the Field Scanalyzer in our recent paper: http://www.publish.csiro.au/FP/pdf/FP16163).

We are currently using the Field Scanalyzer to identify new characteristics of crops that relate to performance, as well as identifying new genetic diversity for existing traits. Outputs from either of these research components can be delivered to breeders. We are screening approximately 400 wheat varieties, but also imaging some oilseed rape and oat plants.

Rothamsted Research

The scanalyzer. Image credit: Rothamsted Research

The big data problem

The Field Scanalyzer at Rothamsted is a world first, so we initially had to develop all the necessary image acquisition protocols and image processing tools, in order to exploit its full capabilities. A number of image processing tools are available; however, they are not suitable for field-grown crops, as they were not developed for complex canopies consisting of hundreds of plants in highly dynamic ambient conditions. The platform can generate up to 100 TB data with a year’s continuous operation (using all of the sensors). That’s why I work with two other post-docs to develop robust computer vision tools to automate the way we extracting quantitative image datasets. We are also validating the accuracy of the values extracted from our images by comparing them with measurements obtained manually.

Approximately 1.5 years have passed since we first began operating the Field Scanalyzer, and we have now optimized all of our image acquisition protocols and have collected a full seasonal dataset. With the good quality images stored in our database, we have developed some robust tools to automatically extract the information about some key growth stages (ear emergence and flowering), as well as quantifying height and the number of some plant organs. We are still just scraping the surface though, and have a list of traits for which we want to develop computer vision tools, in order to automatically analyze them.

Take to the skies: Drones for data collection

Some of my colleagues work with drones (UAVs) to capture information about crop height, plant density (Normalized Difference Vegetation Index), and canopy temperature from large-scale field trials containing 5000 plots. They also fly the UAVs over our Field Scanalyzer site, so we can compare data collected from the higher flying UAV with those collected from the Field Scanalyzer at close proximity. The way we see it, UAVs can image large fields in a very short time (15 min), so if we notice something interesting using the UAV at the large plot-scale, we can put the material under the Field Scanalyzer for high-resolution phenotyping. On the other hand, with the Field Scanalyzer, once we gain a better understanding of which trait/s we need to focus on, when we should be looking at them, and exactly which sensor/s are required to quantify the trait, we can deploy drones with the necessary sensors (once the sensors are portable enough) to collect this information at field-scale and at the appropriate time.

Drones at Rothamsted

Taking to the skies: Drones are used for large-scale phenotyping at Rothamsted. Credit: Rothamsted Research.

The future of phenotyping

I envision that the future of phenotyping technology will focus on reducing the cost and size of cameras/sensors, ultimately increasing their portability and accessibility. This will result in more sophisticated cameras being attached to UAVs (as many of sensors we currently use far out-weigh a UAV’s payload). Parallel to this, research efforts are focusing on developing image processing systems that efficiently extract quantitative information about the crops from acquired images. Together, phenotyping systems such as low-flying UAVs that generate easily interpreted data outputs could be developed, which may be more widely adopted by breeders and farmers to get a deeper insight into their crop’s health and performance.