He attributes this revolution in large part to GCP, saying it “played the role of catalyst. It got things started. It set the foundation. Now we are in a position to do further molecular breeding in chickpeas.”
Led by Pooran, researchers from India, Ethiopia and Kenya worked together not only to develop improved, drought-tolerant chickpeas that would thrive in semiarid conditions, but also to ensure these varieties would be growing in farmers’ fields across Africa within a decade.
The 10-year Generation Challenge Programme, with the goal of improving food security in developing countries, aimed to leave plant genetic assets as an important part of its legacy.
Diagnostic, or informative, molecular markers – which act like ‘tags’ for beneficial genes scientists are looking for – are an increasingly important genetic tool for breeders in developing resilient, improved varieties, and have been a key aspect of GCP’s research.
Chickpeas, ready to harvest.
What is a diagnostic molecular marker?
Developments in plant genetics over the past 10–15 years have provided breeders with powerful tools to detect beneficial traits of plants much more quickly than ever before.
Scientists can identify individual genes and explore which ones are responsible for, or contribute to, valuable characteristics such as tolerance to drought or poor soils, or resistance to pests or diseases.
Once a favourable gene for a target agronomic trait is discovered and located in the plant’s genome, the next step is to find a molecular marker that will effectively tag it. A molecular marker is simply a variation in the plant’s DNA sequence that can be detected by scientists using any of a range of methods. When one of these genetic variants is found close on the genome to a gene of interest (or even within the gene itself), it can be used to indicate the gene’s presence.
To use an analogy, think of a story as the plant’s genome: its words are the plant’s genes, and a molecular marker works like a text highlighter. Molecular markers are not precise enough to highlight specific words (genes), but they can highlight sentences (genomic regions) that contain these words, making it easier and quicker to identify whether or not they are present.
Once a marker is found to be associated with a gene, or multiple genes, and determined to be significant to a target trait, it is designated an informative marker, diagnostic marker or predictive marker. Some simple traits such as flower colour are controlled by one gene, but more complex traits such as drought tolerance are controlled by multiple genes. Diagnostic markers enable plant breeders to practise molecular breeding.
Breeders use markers to predict plant traits
Hard work: a Ugandan bean farmer’s jembe, or hoe.
In the process known as marker-assisted selection, plant breeders use diagnostic molecular markers early in the breeding process to determine whether plants they are developing will have the desired qualities. By testing only a small amount of seed or seedling tissue, breeders are able to choose the best parent plants for crossing, and easily see which of the progeny have inherited useful genes. This considerably shortens the time it takes to develop new crop varieties.
“We use diagnostic markers to check for favourable genes in plants under selection. If the genes are present, we grow the seed or plant and observe how the genes are expressed as plant characteristics in the field [phenotyping]; if the genes are not present, we throw the seed or plant away,” explains Steve Beebe, a leading bean breeder with the International Center for Tropical Agriculture (CIAT) and GCP’s Product Delivery Coordinator for beans.
“This saves us resources and time, as instead of a growing few thousand plants to maturity, most of which would not possess the gene, by using markers to make our selection we need to grow and phenotype only a few hundred plants which we know have the desired genes.”
GCP supported 25 projects to discover and develop markers for genes that control traits that enable key crops, including bean and chickpea, to tolerate drought and poor soils and resist pests and diseases.
Genomic resources, including genetic maps and genotyping datasets, were developed during GCP’s first phase (2004–2008) and were then used in molecular-breeding projects during the second five years of the Programme (2009–2014).
“GCP’s philosophy was that we have, in breeding programmes, genomic resources that can be utilised. Now we are well placed, and we should be able to continue even after GCP with our molecular-breeding programme,” says Pooran.
A small selection of the rice diversity in the International Rice Research Institute gene bank – raw material for the creation of genomic resources.
Markers developed for drought tolerance
With climate change making droughts more frequent and severe, breeding for drought tolerance was a key priority for GCP from its inception.
Different plants may use similar strategies to tolerate drought, for example, having longer roots or reducing water loss from leaves. But drought tolerance is a complex trait to breed, as in each crop a large number of genes are involved.
Wheat, for example, has many traits – each controlled by different genes – that allow the crop to tolerate extreme temperature and/or lack of moisture. Identifying drought tolerance in wheat is therefore a search for many genes. In the particular case of wheat, this search is compounded by its genetic make-up, which is one of the most complex in the plant kingdom.
The difficulty of identifying genes that play a significant role in drought tolerance makes it all the more impressive when researchers successfully collaborate to overcome these challenges. GCP-supported scientists were able to develop and use diagnostic markers in chickpea, rice, sorghum and wheat to breed for drought tolerance. The first new drought-tolerant varieties bred using marker-assisted selection have already been released to farmers in Africa and Asia and are making significant contributions to food and income security.
Tanzanian sorghum farmer.
Markers developed for pests and diseases
A bumper harvest of cassava roots at the International Institute of Tropical Agriculture (IITA) in Nigeria.
Cassava mosaic disease (CMD) is the biggest threat to cassava production in Africa – where more cassava is grown and eaten than any other crop. A principal source of CMD resistance is CMD2, a dominant gene that confers high levels of resistance.
Nigerian GCP-supported researchers worked on identifying and validating diagnostic markers that are associated with CMD2. These markers are being used in marker-assisted selection work to transfer CMD resistance to locally-adapted, farmer-preferred varieties.
In the common bean, GCP-supported researchers identified genes for resistance to pests such as bean stem maggot in Ethiopia, as well as diseases such as the common mosaic necrosis potyvirus and common bacterial blight, which reduce bean quality and yields and in some cases means total crop losses.
Markers developed for acidic and saline soils
Sifting rice in Nepal.
Aluminium toxicity and phosphorus deficiency, caused by imbalanced nutrient availability in acid soils, are major factors in inhibiting crop productivity throughout the world. Aluminium toxicity also exacerbates the effects of drought by inhibiting root growth.
Diagnostic markers for genes that confer tolerance to high levels of aluminium and improve phosphorus uptake were identified in sorghum, maize and rice. The markers linked to these two sets of similar major genes have been used efficiently in breeding programmes in Africa and Asia.
Diagnostic molecular markers are, in their most essential form, data. That means they are easily stored and maintained as data in publicly accessible databases and publications. Breeders can now access the molecular markers developed for various crops through the Integrated Breeding Platform – a web-based one-stop shop for integrated breeding information (including genetic resources), tools and support, which was established by GCP and is now continuing independently following GCP’s close – in order to design and carry out breeding projects.
“We could not have done that much in developing genomic resources without GCP support,” says Pooran. “Now the breeding products are coming; the markers are strengthening our work; and you will see in the next five to six years more products coming from molecular breeding.
“For me, GCP has improved the efficiency of the breeding programme – that is the biggest advantage.”
Common beans are the world’s most important food legume, particularly for subsistence and smallholder farmers in East and Southern Africa. They are a crucial source of protein, are easy to grow, are very adaptable to different cropping systems, and mature quickly.
To some, beans are ‘a near-perfect food’ because of their high protein and fibre content plus their complex carbohydrates and other nutrients. One cup of beans provides at least half the recommended daily allowance of folate, or folic acid – a B vitamin that is especially important for pregnant women to prevent birth defects. One cup also supplies 25–30 percent of the daily requirement of iron, 25 percent of that of magnesium and copper, and 15 percent of the potassium and zinc requirement.
Unfortunately, yields in Africa are well below their potential – between 20 and 30 percent below. The main culprit is drought, which affects 70 percent of Africa’s major bean-producing regions. Drought is especially severe in the mid-altitudes of Ethiopia, Kenya, Malawi and Zimbabwe, as well as across Southern Africa.
“In the past, rains used to be very reliable and people were able to know the right time to plant to meet the rains in critical conditions,” she says. “Now these primary agriculture regions are either not receiving rain for long periods of time, or rains are not falling at the right time.”
Virginia recounts that during the 2011/12 cropping season there were no rains soon after planting, when it is important that beans receive moisture. Such instances can cut bean yields by half.
Steve Beebe in the field.
“Drought is a recurrent problem of rainfed agriculture throughout the world,” says Steve Beebe, a leading bean breeder with the International Center for Tropical Agriculture (CIAT). “Since over 80 percent of the world’s cultivated lands are rainfed, drought stress has major implications for global economy and trade.”
“It’s vital that we develop high-yielding drought-tolerant varieties so as to help farmers, particularly in developing countries, adapt to drought and produce sustained yields for their families and local economies,” says Steve.
For an overview of the work on beans from the perspectives of four different partners, watch our video below, “The ABCs of bean breeding”.
What makes a plant drought tolerant?
The question of what makes a plant drought tolerant is one that breeders have debated for centuries. No single plant characteristic or trait can be fully responsible for protecting the plant from the stress of intense heat and reduced access to water.
“It’s a difficult question to answer for any plant, including beans,” says Steve. “Once you do isolate a trait genetically, it can often be difficult to identify this trait in a plant in the field, for example, identifying the architecture and length of a plant’s roots.”
Phenotyping is an important process in conventional plant breeding. It involves identifying and measuring the presence of physical traits such as seed colour, pod size, stem thickness or root length. Gathering data about a range of such characteristics across a number of different plant lines helps breeders decide which plants to use as parents in crosses and which of the progeny have inherited useful traits.
Root length has long been thought of as a drought-tolerance trait: the longer the root, the more chance it has of tapping into moisture stored deeper in the soil profile.
Given, however, that it is difficult to inspect root length in the field, researchers at CIAT have been exploring other more accessible drought-tolerance traits they can more easily identify and measure. One of these is measuring the weight of the plants’ seeds.
Comparison between varieties in trials of drought tolerant beans at CIAT’s headquarters in Colombia.
Fat beans indicate plants coping with drought stress
“We measure seed weight because we are discovering that under drought stress, drought-tolerant bean varieties will divert sugars from their leaves, stems and pods to their seed,” says Steve. “We call this trait ‘pod filling’, and for us it is the most important drought-tolerance trait to be found over the last several years.”
Finding bean plants with larger, heavier seeds when growing under drought conditions indicates that the plants are coping well, and means farmers’ yields are maintained.
As part of GCP’s Legumes RI, African partners like Virginia have been measuring the seed weight of several advanced breeding lines, which can be used as parents to develop new varieties. These breeding lines have been bred by CIAT and demonstrate this pod-filling process and consequent tolerance of drought.
Although this measurement is relatively cheap and easy for breeders all over the world to do, Steve and his team are interested in finding an even more efficient way to spot plants that maintain full pods under drought.
“We are trying to understand which genes control this trait so we can use molecular-assisted breeding techniques to determine when the trait is present,” says Steve. Having identified several regions of genes related to pod filling, he and his team have developed molecular markers to help breeders identify which plants have these desired genes. “The use of molecular markers in selection significantly reduces the time and cost of the breeding process, making it more efficient. This means that we get improved varieties out to farmers more quickly.”
Bean farmer in Rwanda.
Molecular markers (also known as DNA markers) are used by researchers as ‘flags’ to identify particular genes within a plant’s genome (DNA) that control desired traits, such as drought tolerance. These markers are themselves fragments of DNA that highlight particular genes or regions of genes by binding near them.
To use an analogy, think of a story as the plant’s genome: its words are the plant’s genes, and a molecular marker works like a text highlighter. Molecular markers are not precise enough to highlight specific words (genes), but they can highlight sentences (genomic regions) that contain these words (genes), making it easier and quicker to identify whether or not they are present.
Beans from Rwanda.
Plant breeders can use molecular markers from early on in the breeding process to choose parents for their crosses and determine whether progeny they have produced have the desired trait, based on testing only a small amount of seed or seedling tissue.
“If the genes are present, we grow the progeny and conduct the appropriate phenotyping; if not, we throw the progeny away,” explains Steve. “This saves us resources and time because we need to grow and phenotype only the few hundred progeny which we know have the desired genes, instead of a few thousand progeny, most of which would not possess the gene.”
GCP has supported this foundation work, building on the extensive bean research already done by CIAT dating back to the 1970s, to develop molecular markers not only for drought-tolerance traits such as pod filling, but also for traits associated with resistance to important insect and disease menaces.
“Under drought conditions, plants become more susceptible to pests and diseases, so it was important that we also try to identify and include resistance traits in the drought-tolerant progeny,” says Steve.
Drought is but one plant stressor – diseases and pests wreak havoc too
Common bacterial blight on bean.
The bean diseases that farmers in Ethiopia, Kenya, Malawi and Zimbabwe continually confront are angular leaf spot, bean common mosaic virus, common bacterial blight and rust. Key insect pests are bean stem maggot and aphids.
“We’ve had reports of bean stem maggot and bean common mosaic virus wiping out a whole field of beans,” says Virginia. “Although angular leaf spot and common bacterial blight are not as damaging, they can still reduce yields by over 50 percent.”
Virginia says this is devastating for farmers in Malawi, many of whom only have enough land and money to grow beans to feed their families and sell what little excess there is at market to purchase other necessities.
“This is why we are excited by the prospect of developing not just drought-tolerant varieties, but drought-tolerant varieties with disease and pest resistance as well,” says Virginia.
Virginia’s team in Malawi – along with other breeders in Ethiopia, Kenya and Zimbabwe – are currently using over 200 Mesoamerican and Andean bean breeding lines supplied by CIAT to help breed for drought tolerance and disease and pest resistance. Although many do not yet have the capacity to do molecular breeding in their countries, thanks to advances in plant science it is becoming more feasible and cheaper to outsource molecular breeding stages of the process (see box above).
“With help from GCP and CIAT, we have successfully crossed a line from CIAT with some local varieties to produce plants that are high yielding and resistant to most common bean diseases,” Virginia says.
Malawian farmer Jinny Lemson grows beans to feed her livestock.
Ethiopia’s new bean breeders
Young women sorting beans after a harvest in Ethiopia.
“We’ve [CIAT] hosted several African PhD students here in Colombia and have conducted several workshops in Colombia and Africa too,” says Bodo.
“At the workshops we teach local breeders and technicians how to use genetic tools and markers for advanced breeding methods, phenotyping and data management. The more people there are who can do this work, the quicker new varieties will filter through to farmers.”
Bodo says he has found delivering the training both personally and professionally rewarding, especially “seeing the participants understand the concepts and start using the tools and techniques to develop new lines [of bean varieties] and contribute to the project.”
Daniel started as a GCP-funded Master’s student enrolled at Haramaya University, Ethiopia, evaluating bean varieties with both tolerance to drought and resistance to bean stem maggot. He eventually became the Ethiopian project leader for beans within GCP’s Legumes RI.
“Daniel is currently one of only a handful of bean breeders in Ethiopia who are using molecular-assisted breeding techniques to breed new varieties,” says Bodo. “It’s quite an achievement, especially now that he has taken on the lead role in Ethiopia.”
Buying and selling at a bean market in Kampala, Uganda.
For Daniel, learning about and using the new molecular-breeding techniques has been an exciting new challenge. “The most interesting part of the technology is that it helps us understand what is going on in the plant at a molecular level and lets us know if the crosses we are making are successful and the genes we want are present,” says Daniel. “All this helps improve our efficiency and speeds up the time it takes us to breed and release new varieties for farmers.”
By the end of 2014, Daniel and his team had finished the third year of trials and had several drought-tolerant lines ready for national trials in 2015 and eventual release in 2016.
“The IBP is a really fantastic tool,” says Daniel. “During the course we learnt about the importance of recording clear and consistent phenotypic data, and the IBP helps us to do this as well as store it in a database. It makes it easier to refer to and learn from the past. I’m now trying to pass on the knowledge I’ve learnt as well as create and implement a data-management policy for all plant breeders and technicians in our institute.”
Daniel says the challenge for his institute now is to build further capacity among staff – and to retain it. “At the moment we only have two bean breeders,” says Daniel. “It’s hard to retain research staff in Ethiopia as salaries are very low, so people move on to new, higher paying positions when they get the chance. It’s not unique to Ethiopia, but true of all Africa.”
Bean trials at KALRO in Kenya.
Kenya chasing higher bean yields
Across the border, Kenya has also been facing staffing issues.
And it’s a good thing too, as the country is in need of higher yielding beans to accommodate its population’s insatiable appetite for the crop. Out of the four target African countries, Kenya is the largest bean producer and consumer. As such, the country relies on beans imports from Ethiopia, Malawi, Tanzania and Uganda.
“A lot of families eat beans every day,” says David. “On average, the population eats 14–16 kilograms per person each year, but in western Kenya the average is over 60 kilograms.”
Githeri, a Kenyan staple food made with maize and beans.
Kenyans consume an average total of 400,000 tonnes of beans each year, consistently more than the country produces. Projected trends in population growth indicate that this demand for beans will continue to increase by three to four percent annually.
Even though the area planted to beans has been increasing, David says farmers and breeders need to work together to improve productivity, which is well below where it should be. “The national average yield is 100 kilograms per hectare, which can range from 50 kilograms up to 700 kilograms, depending on whether we experience a drought, or a pest or disease epidemic,” explains David. “The minimum target we should be aiming for is 1,200 kilograms per hectare.”
Such a figure may seem impossible, but David believes that new breeding techniques and the varieties KALRO are producing with the help of CIAT are providing hope that farmers can reach these lofty goals.
“We have several bean lines that are showing good potential to produce higher yields under drought conditions and also have resistance to diseases like rust and mosaic virus,” says David. “They are currently under national trials, and we are confident these will be released to farmers in 2015.”
Varieties fare differently in KALRO bean trials in Kenya.
Maturing bean pods.
“Many subsistence farmers have limited access to good quality bean seeds; they lack knowledge of good crop, pest and disease management; and they have poor post-harvest storage facilities,” says Godwill Makunde, who was previously a breeder at Zimbabwe’s Crop Breeding Institute (CBI) and leader of GCP’s Legumes RI bean project in Zimbabwe.
“Because beans are self-pollinating, which means each crop is capable of producing seed exactly as it was sown, farmers tend to propagate seed on farm,” says Godwill. “While this can be cost effective, it can reduce farmers’ access to higher yielding, tolerant lines, like the ones we are currently producing.”
In none of the partner countries of TLI and TLII are there formal systems for producing and disseminating bean seeds. Godwill and other partners are working with seed companies on developing a sustainable model where both farmers and seed companies can benefit.
Success built on a solid foundation
Field workers tend beans in Rwanda.
A key to the success of the beans component of GCP’s Legumes RI, according to Ndeye Ndack Diop, GCP’s Capacity Building Theme Leader and TLI Project Manager, has been partners’ existing relationships with each other.
BeanCAP released more than 1,500 molecular markers to TLI researchers, which have helped broaden the genetic tools available to developing-country bean breeders.
TLI was also able to leverage and advance previous BeanCAP work and networks. For example, it was through this collaboration that GCP was introduced to LGC Genomics, a company it then worked with on many other crop projects.
To sustain integrated breeding practices beyond the Programme’s close in 2014, GCP established Communities of Practice (CoPs) that are discipline- and commodity-oriented.
“GCP’s CoP for beans has also helped to broaden both the TLI and BeanCAP networks too,” says Ndeye Ndack. “The ultimate goal of the CoPs is to provide a platform for community problem solving, idea generation and information sharing.”
Developing physical capacity
Besides developing human capacity, GCP has also invested in developing infrastructure in Ethiopia, Kenya and Zimbabwe.
SARI now has an irrigation system to enable them to conduct drought trials year round. “We have 12.5 hectares of irrigation now, which we use to increase our efficiency and secure our research,” says Daniel. “We can also increase seed with this irrigation during the off-season and develop early generation seeds for seed producers.”
In Zimbabwe, CBI received specialised equipment that enables them to extract DNA and send it for genotyping in the UK.
Both SARI and CBI also received automatic weather stations from GCP for high-precision climatic data capture, with automated data loading and sharing with other partners in the network.
Delivering the right beans to farmers
Back in Malawi, Virginia says another important facet of the TLII project is that researchers understand what qualities farmers want in their beans. “It’s no use developing higher yielding beans if the farmer doesn’t like the colour, or they don’t taste nice,” she says. “For example, consumers in central Malawi prefer khaki or ‘sugar beans’, which are tan with brown, black or red speckles. While those in southern Malawi tend to prefer red beans. Farmers know this and will grow beans that they know consumers will want.”
Diversity at bean market in Masaka, Uganda.
Breeders in all four countries have been conducting workshops and small trials with farmers to find out this information. In Kenya, David finds farmer participation a great way to promote the work they are doing and show the impact the new drought-tolerant and disease-resistant lines can have.
“Farmers are excited and want to grow these varieties immediately when they see for themselves the difference in yield these new varieties can produce compared to their regular varieties,” says David. “They understand the pressure on them to produce more yields and are grateful that these varieties are becoming more readily available as well as tailored to their needs.”
For Steve, such anecdotes provide him and his collaborators with incentives to continue their quest to discover more molecular markers associated with drought tolerance, post-GCP.
“It’s a testament to everyone involved that we have been able to develop these advanced lines with pod-filling traits using molecular techniques, and make them available to farmers in six years instead of ten,” says Steve.