More than 10,000 kilometers were crossed by scientists, including three states: Bahia, Mato Grosso and Pará. The expeditions made the genetic collection of Embrapa's Vanilla Gene Bank, in Distrito Federal, grow from 70 to 130 specimens of aromatic species. The new collection was created in 2022 with materials collected in Goiás, but the goal is to invest in continuous plant collecting expeditions to make it one of the most representative in the world. There is a potential market for Brazilian vanilla species, especially in haute cuisine. For this purpose, it is important to know and conserve them. Genomic tools are very important in that sense. A booklet was prepared to help organize the production chain for this crop in the country. More than 10,000 kilometers were covered, by roads that cross the Cerrado, the Atlantic Rainforest and the Amazon Rainforest biomes, through several cities and towns in the country. This was the path Embrapa's team followed in search of Brazilian vanilla species. The result: 19 species collected and 60 specimens incorporated into Embrapa's Vanilla Gene Bank, which started in July 2022, in Brasília, DF. After three expeditions in Bahia, Mato Grosso and Pará, the collection went from about 70 to 130 specimens of aromatic species. In the last trip alone, to Pará, 16 species were brought to Brasília, some of which were collected in their natural habitat within the city limits of Belém, Cametá and Tomé-Açu and outskirts, while others came from local orchidists, who already had some of these plants. "This corresponds to about 30% of the species that occur in Brazil", reports the Embrapa Cerrados researcher Fernando Rocha, who was responsible for the activity. “We are assembling one of the most representative collections of Brazilian species in the world, especially those of the groups Vanilla odorata and hostmannii. With each expedition, we expand the known occurrence of any given species and the diversity of Embrapa's gene bank," Rocha adds. Roberto Vieira, a researcher at Embrapa Genetic Resources and Biotechnology and coordinator of the project, says that, the collection will enable the identification of plants with agronomic potential to inform the domestication of vanilla in Brazil and help preserve those species. Product with a lot of potential Brazil has no tradition in the cultivation of vanilla despite having a large diversity of native species in the country. “Brazilian vanilla has the potential to generate income for smallholders. We have prospects that it can have its use expanded in gastronomy and other areas, and can be associated with gastronomic tourism and production diversification, along with other Cerrado products," states Luciano Bianchetti, another researcher at Embrapa Genetic Resources and Biotechnology. Companies and scientists who are interested in the potential of Brazilian species are already in contact with the team. Vanilla is a climbing plant of the orchid family, belonging to the genus Vanilla. It is the only orchid to produce aromatic fruits, the vanilla beans, one of the most appreciated and valued spices in the world, mainly used in the food and cosmetic industries. Nowadays there is a high demand for vanilla products from natural sources in the national and international markets. Brazilian species have unique characteristics and can conquer important markets like haute cuisine. A bean pod can cost about 50 Brazilian reais or more, which opens a new niche for Brazilian farmers. "Many species that occur in Brazil are still unknown in the rest of the world. Its inclusion in promising markets can enhance regional products and generate benefits for rural communities and other interested parties,” states Zenilton de Miranda, an analyst at Embrapa Cerrados. Embrapa's Gene Bank The role of a gene bank is to conserve the largest genetic variability available. It is enriched through efforts to collect genetic material and through exchange with other institutions. Embrapa's Vanilla Gene Bank started its activities less than two years ago and already offers a large representativeness of the species from South America. Despite its economic importance, Vanilla species are underrepresented in germplasm collections around the world Conservation to prevent losses Around the world there are about 110 known species of the genus. Out of those, only 30 to 40 are aromatic and occur only in tropical and subtropical regions of the Americas. Out of the estimated total for the species, about 40 occur in Brazil, and the Amazon and Atlantic rainforest biomes have the greatest diversity. The human presence in the environments where vanilla naturally occurs has endangered the survival of these species. "The ideal is that in situ conservation occurs at the genetic material's place of origin," Rocha warns. However, to prevent specimens from being lost, research institutions search for ways to preserve the planet's biodiversity. Bianchetti, who also participates in the plant collecting expeditions, explains: “Ex situ (out of habitat) conservation of Vanilla species diversity and variability within species are a couple of the goals for Embrapa's Gene Bank.” With the expeditions, materials from about 150 different accessions joined the corporation's collection. The researcher Roberto Vieira justifies the importance of this diversity, especially because there are few materials collected in Brazil: "In order to develop the production chain that is mostly related to gastronomy, it is important to learn the variability between species, including their aromas. For agronomic improvement aimed at productivity, disease resistance and other desirable traits, other materials may have a lot of relevance. As for science, we were able to find materials that had not been recorded in other regions or even described," he adds. In the Atlantic Rainforest, more specifically in the Serra do Conduru State Park in Bahia, where there was an expedition in November 2022, they found Vanilla bicolor, a species with few records outside the Amazon biome, a large population of species of the Vanilla odorata, group, and a species yet to be confirmed. The coordinator explains that for the gene bank, diversity means more safety: “We have collected materials in areas that are not very conserved, as they are subject to different kinds of damage. The most important thing is to have access to those materials, so that they are not lost.” Vanillas from Brazil: a rescue project In two years, five vanilla collecting expeditions were carried out in Brazil. The first two expeditions in Goiás state provided the material that started Embrapa's Gene Bank. Another three took place in the second half of 2022 and in 2023. Even though these types of trips are planned in detail, they are almost an adventure. They include boat trips, walks under the scorching sun and rain, crossing flooded lands, and so many other difficulties that can appear when one is looking for genetic material in natural areas. “This trip was the most complex one because the distances involved have no parallel outside the Amazon," Rocha says. Despite this, the result was very positive: "We found support with Embrapa Eastern Amazon, the Goeldi Museum and local collectors, such as orchidist Tiago Carneiro, who turned this trip into the richest one in terms of collected species." DNA analysis for species identification Identifying a vanilla species is no easy task. Although there is a difference between the leaves of the species, confirmation generally depends on the analysis of their flowers. The problem is that the time from seedling to blooming can take up to three years, according to Embrapa researcher Marília Pappas. The use of genomic tools could make such activity easier and faster. The researcher collects leaves from all the samples that reach the Gene Bank and extracts their DNA, a structure that carries the genetic information of an organism. That is how she ensures there is a backup of the genetic information of those materials in Embrapa's Plant DNA Bank, of which she is a curator, a safety net in case the plant does not survive in the collection. "DNA analyses not only can help us learn more about the genetic diversity of the materials in the Gene Bank and thus about the diversity in the areas that were sampled, but also accelerates species identification", she explains. That is the case of some plants that come from the fields without species confirmation based on their morphology. Genetic analysis can benefit those who are interested in cultivating vanilla, as it ensures buyers that the seedling they are acquiring is actually of the intended species, and allows seedling suppliers to certify their production. The genomic methodologies used for vanillas are the same as those applied to commercially cultivated species, such as commodities. "It is common to have little or no genomic information available for species of little economic appeal, which are known as neglected species. Therefore, it is necessary to generate the initial information and thus build a database to support research in several areas”, she points out In addition, Pappas is looking for molecular markers to identify Brazilian vanilla species early. "Our first goal is to obtain a panel of molecular markers for the many different national species, which will allow their early identification, when they are still in the seedling stage. Hence it will be possible to generate information on the diversity of natural populations and significantly contribute to species conservation and to the strengthening of the production chain," she argues. From Brasília to Mato Grosso Out of a total of about 40 Brazilian Vanilla species, less than half (between 15 and 17) bear aromatic fruits. They are mostly spread across the Amazon and Atlantic Rainforest biomes. While the north of the state of Mato Grosso belongs to the Cerrado biome, it borders the south of the Brazilian Amazon. "The choice of region for the plant collecting expedition was Itaúba and neighboring towns, as it is transitional between two biomes and houses a lot of both biodiversities, and thus usually offers high species diversity", Rocha explains. The premise was proven correct. That expedition resulted in the largest collection of 2022, which demonstrates that the site can be considered a center of diversity for the genus. "In a very small area, covering a few town limits, 16 accessions belonging to probably eight or nine species were collected in just five days," Rocha says. Despite the increased representativeness of Embrapa's Vanilla Gene Bank, much remains to be done. “In a short span of time, we managed to collect a significant amount of the species that are known in Brazil. But when we look at the data, both from the literature and herbaria as well as those we collected, we have noticed that there is a large sampling gap in the countryside of Brazil, especially in southern Pará and Amazonas, combined with Amapá”, the Embrapa Cerrados researcher assesses. Challenges of setting up cultivation systems The expeditions actively sought local growers. In all the Brazilian states they crossed – Goiás, Bahia, Mato Grosso and Pará –, visited properties had difficulties in the production of Brazilian vanillas. Some grow them under screens, others, in agroforestry systems. But there are common issues: inadequate soil moisture and plant densification, several diseases caused by fungi and viruses, and pest attacks. Those field observations are important for research as, in addition to preserving the genus, there is the aim to develop a system for the production of Brazilian vanillas. “This stage [of contact with people who work with vanilla] is essential to structure the production chain for Brazilian vanilla species. Nowadays most of the exploitation of native species, especially in Goiás, takes place through extractivism”, the researcher Roberto Vieira explains. In general, the information available on the cultivation of such orchids is in foreign languages, which makes it difficult for farmers to cultivate the species. Hence the relevance of the first of the project's results for the organization of the production chain for the crop: the booklet Cultivo de Baunilha – Práticas Básicas [Vanilla Cultivation – Basic Practices]. The publication gathers information on good practices in vanilla cultivation and management in Brazil to obtain quality fruits, based on knowledge of cultivation systems, phytosanitary aspects, flowering and pollination, post-harvest processes, and processing. The project also includes training for farmers, gastronomy professionals and quilombola (escaped slave-descending) communities. Vieira emphasizes the importance of disseminating such knowledge: "Studying their cultivation and societal awareness are fundamental for the conservation of vanillas. Management is crucial for the establishment of the Brazilian vanilla production chain, which includes the set-up of nurseries to produce seedlings for different markets and for family farming." From trip prep to vanilla materials The success of the trips searching for species depends on good planning. The first step is to establish a geographic range. The goal can be searching for a specific species or one that is about to be lost. In the case of the expeditions by Embrapa's team, they actively sought areas where there is a concentration of different species. For that purpose, they consulted with national and foreign herbaria, places where dehydrated plants are preserved with information on the place from which they were collected. “It is possible to visualize all the plants that have been historically collected in Brazil in order to have an idea of where they are and the regions with the highest number of species, which is essential to define the trip itinerary”, the researcher Fernando Rocha says. With a good literature review, it is possible to trace the itinerary with possible collection points. The person responsible for the vanilla collection activity adds: "In addition, it is essential to establish contacts with people who know the species, with researchers who have already performed collections in the intended region, and with guides with proven experience in the subject".