Isidro Ovando-Medina

Genetic Diversity of Jatropha curcas in Southern Mexico

The importance of the Euphorbiaceous plant Jatropha curcas L. lies in its high-quality seed oil, ideal for the manufacture of biodiesel. As a result, its extensive cultivation has already reached several million hectares in Asia, Africa and Latin America, and implies certain challenges since it is not yet a fully domesticated plant, and with limited information on the agronomic processes and selected varieties for its cultivation. In addition, little is known about the biology, ecology, genetic diversity and geographic origin of its populations. Despite the growing body of knowledge on J. curcas, there are only a few studies on phenotypic and genotypic diversity in populations of Mexico, from where it is probably native. The greatest extension of J. curcas sown in Mexico is located in the state of Chiapas, for that reason this chapter describes diversity of populations of this state and other sites in Southern Mexico. To broadly understand the diversity of populations, our group has used two types of markers: direct adaptive value characteristics, such as floral characters and fatty acids in the seed, and neutral molecular markers such as AFLP and microsatellites. The results of the chemical studies showed that the content of seed oil in accessions ranged from 8.02 % to 54.28 %, with the proportion of unsaturated fatty acids varying from 74.5 % to 83.7 %. A discriminant analysis separated populations according to their geographical origin, which was verified with a Mantel test. Using the Monmonier’s algorithm, two genetic barriers were identified between the populations. The results are discussed according to the putative evolutionary significance of the seeds fatty acid composition for this tropical species. Studies on floral anatomy confirmed a large variation among Mexican J. curcas and revealed the existence of plants exclusively with female inflorescences, which could be useful in the development of future plantations. Molecular studies showed that Mexican J. curcas is moderately structured in subpopulations, but with a high diversity within populations. A conglomerate analysis revealed the highest coefficient of dissimilarity based on accessions reported so far. The existence of genetic barriers isolating some populations was confirmed with molecular data. The results are discussed with reference to the possible ancestry of the populations. Finally, we highlight the relevance of some of our findings in the scientific and productive realms.

Reproductive biology of the biofuel plant Jatropha curcas in its center of origin

In this work, we studied the main characteristics of flowering, reproductive system and diversity of pollinators for the biofuel plant Jatropha curcas (L.) in a site of tropical southeastern Mexico, within its center of origin. The plants were monoecious with inflorescences of unisexual flowers. The male flowers produced from 3062–5016 pollen grains (266–647 per anther). The plants produced fruits with both geitonogamy and xenogamy, although insect pollination significantly increased the number and quality of fruits. A high diversity of flower visiting insects (36 species) was found, of which nine were classified as efficient pollinators. The native stingless bees Scaptotrigona mexicana (Guérin-Meneville) and Trigona (Tetragonisca) angustula (Latreille) were the most frequent visitors and their presence coincided with the hours when the stigma was receptive. It is noteworthy that the female flowers open before the male flowers, favoring xenogamy, which may explain the high genetic variability reported in J. curcas for this region of the world.

Genetic identification of Theobroma cacao L. trees with high Criollo ancestry in Soconusco, Chiapas, Mexico.

Criollo-type cacao trees are an important pool of genes with potential to be used in cacao breeding and selection programs. For that reason, we assessed the diversity and population structure of Criollo-type trees (108 cultivars with Criollo phenotypic characteristics and 10 Criollo references) using 12 simple sequence repeat (SSR) markers. Cultivars were selected from 7 demes in the Soconusco region of southern Mexico. SSRs amplified 74 alleles with an average of 3.6 alleles per population. The overall populations showed an average observed heterozygosity of 0.28, indicating heterozygote deficiency (average fixation index F = 0.50). However, moderate allelic diversity was found within populations (Shannon index for all populations I = 0.97). Bayesian method analysis determined 2 genetic clusters (K = 2) within individuals. In concordance, an assignment test grouped 37 multilocus genotypes (including 10 references) into a first cluster (Criollo), 54 into a second (presumably Amelonado), and 27 admixed individuals unassigned at the 90% threshold likely corresponding to the Trinitario genotype. This classification was supported by the principal coordinate analysis and analysis of molecular variance, which showed 12% of variation among populations (FST = 0.123, P < 0.0001). Sampled demes sites (1- 7) in the Soconusco region did not show any evidence of clustering by geographic location, and this was supported by the Mantel test (Rxy = 0.54, P = 0.120). Individuals with high Criollo lineage planted in Soconusco farms could be an important reservoir of genes for future breeding programs searching for fine, taste, flavor, and aroma cocoa.

Fatty acid diversity is not associated with neutral genetic diversity in native populations of the biodiesel plant Jatropha curcas L.

Jatropha curcas L. (Euphorbiaceae) is a shrub native to Mexico and Central America, which produces seeds with a high oil content that can be converted to biodiesel. The genetic diversity of this plant has been widely studied, but it is not known whether the diversity of the seed oil chemical composition correlates with neutral genetic diversity. The total seed oil content, the diversity of profiles of fatty acids and phorbol esters were quantified, also, the genetic diversity obtained from simple sequence repeats was analyzed in native populations of J. curcas in Mexico. Using the fatty acids profiles, a discriminant analysis recognized three groups of individuals according to geographical origin. Bayesian assignment analysis revealed two genetic groups, while the genetic structure of the populations could not be explained by isolation‐by‐distance. Genetic and fatty acid profile data were not correlated based on Mantel test. Also, phorbol ester content and genetic diversity were not associated. Multiple linear regression analysis showed that total oil content was associated with altitude and seasonality of temperature. The content of unsaturated fatty acids was associated with altitude. Therefore, the cultivation planning of J. curcas should take into account chemical variation related to environmental factors.

Sex expression and floral diversity in Jatropha curcas: a population study in its center of origin

Sex expression and floral morphology studies are central to understand breeding behavior and to define the productive potential of plant genotypes. In particular, the new bioenergy crop Jatropha curcas L. has been classified as a monoecious species. Nonetheless, there is no information about its reproductive diversity in the Mesoamerican region, which is considered its center of origin and diversification. Thus, we determined sex expression and floral morphology in J. curcas populations from southern Mexico and Guatemala. Our results showed that most of J. curcas specimens had typical inflorescences with separate sexes (monoecious); meanwhile, the rest were atypical (gynoecious, androecious, andromonoecious, androgynomonoecious). The most important variables to group these populations, based on a discriminant analysis, were: male flower diameter, female petal length and male nectary length. From southern Mexico “Guerrero” was the most diverse population, and “Centro” had the highest variability among the populations from Chiapas. A cluster analysis showed that the accessions from southern Mexico were grouped without showing any correlation with the geographical origin, while those accessions with atypical sexuality were grouped together. To answer the question of how informative are floral morphological traits compared to molecular markers, we perform a Mantel correlation test between the distance matrix generated in this study and the genetic distance matrix (AFLP) previously reported for the same accessions. We found significant correlation between data at the level of accessions. Our results contribute to design genetic improvement programs by using sexually and morphologically contrasting plants from the center of origin.

Unique haplotypes of cacao trees as revealed by trnH-psbA chloroplast DNA

Cacao trees have been cultivated in Mesoamerica for at least 4,000 years. In this study, we analyzed sequence variation in the chloroplast DNA trnH-psbA intergenic spacer from 28 cacao trees from different farms in the Soconusco region in southern Mexico. Genetic relationships were established by two analysis approaches based on geographic origin (five populations) and genetic origin (based on a previous study). We identified six polymorphic sites, including five insertion/deletion (indels) types and one transversion. The overall nucleotide diversity was low for both approaches (geographic = 0.0032 and genetic = 0.0038). Conversely, we obtained moderate to high haplotype diversity (0.66 and 0.80) with 10 and 12 haplotypes, respectively. The common haplotype (H1) for both networks included cacao trees from all geographic locations (geographic approach) and four genetic groups (genetic approach). This common haplotype (ancient) derived a set of intermediate haplotypes and singletons interconnected by one or two mutational steps, which suggested directional selection and event purification from the expansion of narrow populations. Cacao trees from Soconusco region were grouped into one cluster without any evidence of subclustering based on AMOVA (FST = 0) and SAMOVA (FST = 0.04393) results. One population (Mazatán) showed a high haplotype frequency; thus, this population could be considered an important reservoir of genetic material. The indels located in the trnH-psbA intergenic spacer of cacao trees could be useful as markers for the development of DNA barcoding.

Association Between SSR Molecular Markers and Chemical and Sensory Traits of Cacao Samples Using Multiple Regression Analysis

The sensory properties of cacao beans are linked to the chemical composition of the seeds, and both characteristics are the partial results of its allelic composition. Therefore, it is useful to search for molecular markers associated with these traits. We perform multiple regression analysis to associate previously generated data of alleles generated with 12 SSR (of cultivated cacao trees) with data obtained from chemical and sensory characterization (of beans) of plants grown in the southern region from Mexico. When the association was significant, the mathematical models for predictive purposes were proposed. All phenotypic traits evaluated showed equations with setting values R2 > 0.5. All chemical characters tested have a significant association with at least two alleles (P < 0.05). In addition, the fat content was associated with six molecular markers (mTcCIR03209, mTcCIR12188, mTcCIR19286, mTcCIR07150, mTcCIR19310). The most common allele was mTcCIR12188, which was associated with the contents of eicosanoic acid, moisture, fat and total polyphenols content. The mTcCIR28362 allele is associated with sensory characters bitterness, musty odor, and roasted odor. These alleles could be useful as molecular markers of chemical and sensory characteristics of cacao samples.

Production of cytotoxic compounds in dedifferentiated cells of Jatropha curcas L. (Euphorbiaceae)

This study addresses the in vitro culture as an alternative to obtain compounds with cytotoxic activity from the medicinal plant Jatropha curcas (Euphorbiaceae). We determined the presence of cytotoxic compounds in both whole plants and dedifferentiated cells. We evaluated the effect of auxin, cytokinins and light on callus induction in cotyledon explants. We found that the most effective combination to induce callus was the auxin 2,4-D (5 mM) with the cytokinin 6-BAP (2.5 mM), on Murashige-Skoog medium in darkness. We compared the callogenic potential among accessions from different geographic origins, finding that ARR-251107-MFG7 is most prone to form callus. The roots of J. curcas grown in field produced a compound chromatographically similar to the cytotoxic diterpene jatrophone. The profile of compounds extracted from the dedifferentiated cells was similar to that of the whole plant, including a relatively abundant stilbene-like compound. This study contributes to the future establishment of protocols to produce anti-cancer compounds from J. curcas cultivated in vitro.