Molecularly Imprinted Polymer-Coated Probe Electrospray Ionization Mass Spectrometry Determines Phorbol Esters and Deoxyphorbol Metabolites in Jatropha curcas Leaves.

In this study, a molecularly imprinted polymer-coated probe electrospray ionization mass spectrometry (MIPCPESI-MS) method was developed for detection of phorbol esters (PEs) and deoxyphorbol metabolites in Jatropha curcas leaves. Such an approach was established by sticking on a metallic needle a molecularly imprinted polymer to particularly design a MIP-coated probe for selective sampling and ionization of PEs and deoxyphorbol metabolites. By a subsequent application of a high voltage and methanol, as spray solvent, ESI was generated for direct and rapid analysis under ambient and open-air conditions. MIP-coated probe exhibited a high sampling capacity of the PEs and its metabolites in methanolic extracts of J. curcas leaves compared with the non-imprinted polymer (NIP)-coated probe. MIPCPESI-MS allowed the detection of phorbol 12,13-diacetate (PDA) from J. curcas leaves with minimal sample preparation, and with detection limit and quantification reaching 0.28 µg/mL and 0.92 µg/mL, respectively. Also, good linearity was obtained with R2 > 0.99 and precision and accuracy values between 4.06-13.49% and - 1.60 to - 15.26%, respectively. The current method was successfully applied to screening methanolic extracts of six different J. curcas leaf genotypes (three toxic and three non-toxic). PDA and three PE deoxyphorbol metabolites were identified only from toxic genotypes, in which PDA was determined with concentration ranging from 222.19 ± 23.55 to 528.23 ± 19.72 µg/g. All these findings support that the MIPCPESI-MS method developed here has a high potential for the analysis of PEs in plant extracts enabling differentiation of toxic and non-toxic genotypes earlier in the leaves.

Perennial plants for biofuel production: bridging genomics and field research.

Development of dedicated perennial crops has been indicated as a strategic action to meet the growing demand for biofuels. Breeding of perennial crops,however, is often time- and resource-consuming. As genomics offers a platform from which to learn more about the relationships of genes and phenotypes,its operational use in the context of breeding programs through strategies such as genomic selection promises to foster the development of perennial crops dedicated to biodiesel production by increasing the efficiency of breeding programs and by shortening the length of the breeding cycles.

Effects of Jatropha curcas oil in Lactuca sativa root tip bioassays.

Jatropha curcas L. (Euphorbiaceae) is important for biofuel production and as a feed ingredient for animal. However, the presence of phorbol esters in the oil and cake renders the seeds toxic. The toxicity of J. curcas oil is currently assessed by testing in animals, leading to their death. The identification of toxic and nontoxic improved varieties is important for the safe use of J. curcas seeds and byproducts to avoid their environmental toxicity. Hence, the aim of this study was to propose a short-term bioassay using a plant as a model to screen the toxicity of J. curcas oil without the need to sacrifice any animals. The toxicity of J. curcas oil was evident in germination, root elongation and chromosomal aberration tests in Lactuca sativa. It was demonstrated that J. curcas seeds contain natural compounds that exert phyto-, cyto- and genotoxic effects on lettuce, and that phorbol esters act as aneugenic agents, leading to the formation of sticky chromosomes and c-metaphase cells. In conclusion, the tests applied have shown reproducibility, which is important to verify the extent of detoxification and to determine toxic doses, thus reducing the numbers of animals that would be used for toxicity tests.

Analysis of morpho-agronomic and climatic variables in successive agricultural years provides novel information regarding the phenological cycle of Jatropha in conditions of the Brazilian cerrado.

Phenological studies can provide information that enables the understanding of the dynamics of plants and how these dynamics are related to the biotic and abiotic environment. In order to study the phenological phases of Jatropha during two agricultural years, agronomic and climatic variables, such as temperature and rainfall, were evaluated. Data for each variable in each year and each genotype were subjected to analysis of variance (ANOVA) and the differences were tested at 5% probability by F test. In addition, the correlation of growth behavior and reproductive development of two Jatropha accessions (CNPAE-102 and CNPAE-169) as a function of time elapsed after the start of the phenological cycle with climatic variables were analyzed through Pearson's correlation. It was found that: (i) the resuming of plant growth by producing new branches and flowers of both genotypes coincides with the start of the rainy season, (ii) the flowering may be related to the increase in temperature and rainfall; (iii) the number of inflorescences per plant and number of female flowers determine the number of green fruits, (iv) the environmental changes are responsible for the delimitation of phenophases; and finally that (v) the responses to phenological changes are genotype-dependent.