Startup creates transgenic sugarcane with enhanced pest resistance

By Suzel Tunes*  |  FAPESP Innovative R&D – The sugarcane borer Diatraea saccharalis is the worst threat to this crop in Brazil, causing annual losses estimated at about a billion dollars. The moth’s larvae are only about 20 mm long but can ruin a field of sugarcane by tunneling through the stems. To combat it, PangeiaBiotech uses even smaller organisms and genetic engineering tools. The startup based in Campinas, in the state of São Paulo, develops genetically modified (GM) sugarcane varieties that associate expression of two bioinsecticidal proteins from the bacterium Bacillus thuringiensis (Bt) with a gene from another microorganism, Agrobacterium sp., that increases the plant’s tolerance of the herbicide glyphosate. Bt genes have been used in many GM crops to control pests biologically for over two decades.

This BtRR technology, as it is known, was developed with support from FAPESP’s Innovative Research in Small Business Program (PIPE), the Brazilian Company of Research and Industrial Innovation (EMBRAPII), and the Brazilian Agricultural Research Corporation (EMBRAPA), which is conducting tests on its experimental plots in Brasília. The next step is to find commercial partners interested in licensing the technology. The startup plans to bring its first variety to market by the time the 2022-23 crop is planted. “We hope to have 20% of Brazil’s sugarcane acreage growing our GM varieties by 2030,” says agricultural engineer Paulo Cezar de Lucca, who designed the project and founded the firm in 2015.

The firm, which is hosted by INCAMP, the University of Campinas’s tech incubator, is not the first developer of GM sugarcane in Brazil. The first GM variety, CTC20BT, was produced in the laboratories of the Sugarcane Technology Center (CTC), maintained by growers and mill owners in Piracicaba, state of São Paulo. CTC20BT was approved in 2017 by CTNBio, Brazil’s biosafety watchdog. CTC won approval for its second GM variety (CTC9001BT) the following year. Both also use a Bt gene to express a Cry protein. Cry proteins are bioinsecticidal substances secreted in crystal form by Bt. When they are ingested by insect pests, they bind to gut receptors, causing fatal damage to the digestive system.

PangeiaBiotech’s novel varieties have taken the technological evolution of sugarcane a step forward by using two different Cry proteins. “Double transgenics has been a feature of crops such as corn and soybeans for some time. Now we’re bringing it to sugarcane,” de Lucca says. For agricultural engineer Hugo Molinari, a researcher at EMBRAPA Agroenergy Unit and a participant in the project, using two proteins with insecticidal properties makes the technology more durable and reduces the likelihood that pests will develop resistance.

The varieties developed by the startup also contain the glyphosate-tolerant gene cp4-epsps from Agrobacterium, which is naturally found in soil. “Resistance to glyphosate in sugarcane is an innovation. Growers can reduce the amount of agrochemicals they use. No sugarcane varieties on the market are resistant to both the borer and glyphosate,” de Lucca says

Herbicide is typically applied between the rows of sugarcane, taking care not to damage the crop. This operation requires the use of tractors and is costly (mainly owing to the cost of diesel fuel) and time-consuming. “If herbicide-resistant sugarcane is used, it can be sprayed by airplane, which saves fuel,” de Lucca says. It also saves herbicide, according to Molinari.

Another innovation under development at PangeiaBiotech is a GM variety resistant not just to the borer and glyphosate but also to Sphenophorus levis, a billbug in the weevil family. “Sugarcane is now harvested mechanically without burning, so billbugs and spittlebugs [Mahanarva fimbriolata] aren’t destroyed by fire as they used to be. They live in the straw and multiply,” Molinari says. The annual crop losses caused by S. levis in Brazil are estimated at some USD 372 million, and the chemical and biological weapons available are not very effective.

Bringing its BtRR varieties to market will consolidate a change in PangeiaBiotech’s business model. According to de Lucca, the firm began by offering genetic engineering services based on the US concept of a plant transformation facility, which did not exist in Brazil at the time. “This project has made strides, and currently services 25 research centers in Brazil,” he says. “They send us the gene of interest, and after four months we send back the modified plant. This enables researchers to focus on discovering new genes. They can quickly see how their theories work out in practice.” The firm supplies GM tobacco, tomato and corn, as well as sugarcane.

The double transgenics sugarcane project and reformulated business plan arose in 2017 from the relationship with EMBRAPA Agroenergy, then one of the firm’s customers. “Our partnership with EMBRAPA enabled us to take a great leap forward. We can now produce our own varieties,” says de Lucca, who also plans to continue providing services.

Other customers include the University of Campinas’s Molecular Biology and Genetic Engineering Center (CBMEG), the Sugarcane Center at the Campinas Institute of Agronomy (IAC), and the Inter-University Network for the Development of the Sugar and Energy Sector (RIDESA), with ten federal universities.

Monalisa Sampaio Carneiro, a professor at the Federal University of São Carlos (UFSCar), one of the ten, claims that RIDESA is responsible for developing more than half the genetically improved varieties grown in Brazil, and now uses the plant transformation services provided by PangeiaBiotech to obtain GM versions of its varieties. “By receiving plants that have already been genetically transformed by PangeiaBiotech, researchers and sugarcane breeders can save up to two years of hard work,” she says.

José Antônio Bressiani, an agricultural engineer who heads the farmer-oriented activities of Brazilian biotech company GranBio, commissioned PangeiaBiotech to develop a GM sugarcane variety currently in field testing.

Another group of specially bred varieties is known as “energy cane”. These are hardier and have a higher fiber content than other varieties. Their straw and bagasse are used to produce second-generation ethanol. GranBio plans to launch a GM variety of energy cane that will be resistant to borers and herbicides. PangeiaBiotech is participating in this project. GranBio is also developing another GM variety designed to be drought-resistant and have more biomass, also with support from FAPESP.

For agricultural engineer Gonçalo Amarante Guimarães Pereira, affiliated with the University of Campinas’s Institute of Biology (IB-UNICAMP), startups like PangeiaBiotech can play a key role in the development of the Brazilian sugar and energy industry, focusing on the global market for biofuels. “Startups are small and nimble, with significant capacity to innovate,” he says.

Pereira has headed the university’s Genomics and Gene Expression Laboratory since its inception in 1997 and was GranBio’s chief scientist between 2012 and 2016. In recent years, he says, genetic improvement of sugarcane has not guaranteed the leaps in yield seen in other crops, such as corn, soybeans, and wheat, after the development of GM varieties. “There’s a limit to traditional genetic improvement. It takes about ten years to breed a new variety, while a novel variant of a microorganism capable of attacking it can pop up in a matter of days,” he says, adding that hardier and more productive plants can be developed via transgenics faster than via conventional breeding.

Although hundreds of sugarcane varieties have been created to suit different climate and soil conditions, some 20 dominate the market and yields have remained more or less stable in recent years. PangeiaBiotech expects to contribute to a change in this situation a quarter of a century after the first Bt corn and cotton varieties were bred in the US.

This lag can be explained commercially and scientifically, according to the researchers. For Molinari, sugarcane is an important energy source but market demand for it is not sufficient to attract investment in research by multinationals. “Sugarcane isn’t global. It’s grown only in the tropics. It’s small compared to the gigantic market for soybeans or wheat,” he says.

The scientific rationale has to do with complexity. “The sugarcane genome is much larger and more complex than those of other plants,” says Michael dos Santos Brito, a biologist at the Federal University of São Paulo’s Institute of Science and Technology (ICT-UNIFESP). The most complete sequencing of the genome to date was not concluded until late 2019, mapping 373,869 genes, or 99.1% of the total.

The sequencing resulted from the FAPESP Bioenergy Research Program (BIOEN), launched in 2008 to stimulate bioenergy production in Brazil. Also under the auspices of BIOEN, Brito is conducting a project to identify and characterize novel plant promoters for sugarcane. Promoters are DNA sequences that turn genes on or off. He plans to create a database for use in future research as part of the project. “Sugarcane isn’t a simple plant,” he says. “Research on sugarcane requires a lot of resources. We must make good use of the know-how developed so far, putting us at the forefront of global R&D in this field.”

* Pesquisa FAPESP magazine