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IntroductionA combination of factors including migration, climate change and the encroachment of growing cities are associated with the extinction of species and a reduction in biodiversity. In Mexico maize landraces are an important source of germplasm that could be invaluable for future breeding programs in response a changing environment. Efforts to develop conservation strategies are hampered by the costs of housing and maintaining large germplasm collections. Effective criteria are therefore needed to identify rare genotypes that are in danger of being lost and to define minimal cost-effective core collections.MethodsA large-scale genotyping analysis of Mexican maize landraces was carried out using 14 SSR markers spread across the 10 maize chromosomes. DNA was extracted from 30 individual plants of each accession and 3 bulks containing 220ng of DNA from each of 10 individual plants were prepared. Amplification reactions were carried out individually on each bulk sample and SSRs were detected using an automatic sequencer. Binary scores were used to produce phylogenies and a rareness index Ri was developed to identify a minimal collection of accessions.ResultsA total of 40,140 individual plants from accessions obtained in 14 different Mexican states and covering 47 maize races were analyzed. Accessions from Northern and Central-Southern states could be distinguished based on genotype and accessions from Michoacán state were found to be distinct to all other accessions. A minimal collection of 56 accessions that covered all identified alleles and included rare allele combinations was determined.DiscussionThe strategy employed successfully distinguished accessions from different regions of Mexico and identified rare germplasm and offers a cost effective method to determine minimal germplasm collections for conservation in situ or ex situ. The strategy could easily be adapted for use in other native crop species such as common bean, chili pepper and tomato among others.

Gene co-expression networks are powerful tools to understand functional interactions between genes. However, large co-expression networks are difficult to interpret and do not guarantee that the relations found will be true for different genotypes. Statistically verified time expression profiles give information about significant changes in expressions through time, and genes with highly correlated time expression profiles, which are annotated in the same biological process, are likely to be functionally connected. A method to obtain robust networks of functionally related genes will be useful to understand the complexity of the transcriptome, leading to biologically relevant insights. We present an algorithm to construct gene functional networks for genes annotated in a given biological process or other aspects of interest. We assume that there are genome-wide time expression profiles for a set of representative genotypes of the species of interest. The method is based on the correlation of time expression profiles, bound by a set of thresholds that assure both, a given false discovery rate, and the discard of correlation outliers. The novelty of the method consists in that a gene expression relation must be repeatedly found in a given set of independent genotypes to be considered valid. This automatically discards relations particular to specific genotypes, assuring a network robustness, which can be set a priori. Additionally, we present an algorithm to find transcription factors candidates for regulating hub genes within a network. The algorithms are demonstrated with data from a large experiment studying gene expression during the development of the fruit in a diverse set of chili pepper genotypes. The algorithm is implemented and demonstrated in a new version of the publicly available R package “Salsa” (version 1.0).

Abstract Background RNA-Seq is the preferred method to explore transcriptomes and to estimate differential gene expression. When an organism has a well-characterized and annotated genome, reads obtained from RNA-Seq experiments can be directly mapped to that genome to estimate the number of transcripts present and relative expression levels of these transcripts. However, for unknown genomes, de novo assembly of RNA-Seq reads must be performed to generate a set of contigs that represents the transcriptome. These contig sets contain multiple transcripts, including immature mRNAs, spliced transcripts and allele variants, as well as products of close paralogs or gene families that can be difficult to distinguish. Thus, tools are needed to select a set of less redundant contigs to represent the transcriptome for downstream analyses. Here we describe the development of Compacta to produce contig sets from de novo assemblies. Results Compacta is a fast and flexible computational tool that allows selection of a representative set of contigs from de novo assemblies. Using a graph-based algorithm, Compacta groups contigs into clusters based on the proportion of shared reads. The user can determine the minimum coverage of the contigs to be clustered, as well as a threshold for the proportion of shared reads in the clustered contigs, thus providing a dynamic range of transcriptome compression that can be adapted according to experimental aims. We compared the performance of Compacta against state of the art clustering algorithms on assemblies from Arabidopsis, mouse and mango, and found that Compacta yielded more rapid results and had competitive precision and recall ratios. We describe and demonstrate a pipeline to tailor Compacta parameters to specific experimental aims. Conclusions Compacta is a fast and flexible algorithm for the determination of optimum contig sets that represent the transcriptome for downstream analyses.

Chili pepper (Capsicum annuum) is one of the most important crops worldwide. Its fruits contain metabolites produced over the maturation process like capsaicinoids and carotenoids. This metabolic process produces internal changes in flavor, color, texture, and aroma in fruits to make them more attractive for seed dispersal organisms. The chiltepin (C. annuum L. var. glabriusculum) is a wild variety of the C. annuum L. species that is considered a source of genetic resources that could be used to improve the current chili crops. In this study, we performed a transcriptomic analysis on two fruit maturation stages: immature stage (green fruit) and mature stage (red fruit) of a wild and a cultivated pepper variety. We found 19,811 genes expressed, and 1,008 genes differentially expressed (DEGs) in at least one of the five contrast used; 730 DEGs were found only in one contrast, and most DEGs in all contrasts were downregulated. GO enrichment analysis showed that the majority of DEGs are related to stress responses. KEGG enrichment analysis detected differences in expression patterns in metabolic pathways related to phenylpropanoid biosynthesis, secondary metabolites, plant hormone signal transduction, carotenoid biosynthesis and sesquiterpenoid and triterpenoid biosynthesis. We selected 105 tomato fruit ripening-related genes, and found 53 pepper homologs differentially expressed related to shape, size, and secondary metabolite biosynthesis. According to the transcriptome analysis, the two peppers showed very similar gene expression patterns; differences in expression patterns of genes related to shape, size, ethylene and secondary metabolites biosynthesis suggest that changes produced by domestication of chilli pepper could be very specific to the expression of genes related to traits desired in commercial fruits.

Microalgae have great potential as biofactories for producing a wide variety of valuable products. Under stress conditions, it is well-known that microalgae accumulate various organic compounds. However, there is little information on the response of the diatom Chaetoceros muelleri to different nitrogen concentrations. In the present study, C. muelleri was grown in medium containing different concentrations of nitrogen (NaNO3; f: 1.76 mM, f/4: 0.44 mM, f/10: 0.18 mM, and 2f: 3.53 mM) and their proximal composition, fatty acid profile, and gene expression (AMT1, PTD9, GltD, DGAT2, and NRT2) compared. The results showed no significant differences in biomass content among nitrogen treatments. Treatment f/10 significantly increased accumulation of lipids and carbohydrates but lowered the concentration of proteins. This treatment also produced shorter-chain fatty acids and a low percentage of polyunsaturated fatty acids, demonstrating the potential of this medium for growing microalgae with a suitable fatty acid profile for biodiesel production. Furthermore, DGAT2 and NRT2 expression was highest at lower nitrogen concentrations, suggesting these genes could be important for modulating lipid biosynthesis in response to nitrogen limitation. Keywords: Chaetoceros muelleri, Nitrogen stress, Fatty acid methyl esters, Biochemical composition, Gene expression

Diatoms are the most abundant group of phytoplankton, and their success lies in their significant adaptation ability to stress conditions, such as nutrient limitation. Phosphorus (P) is a key nutrient involved in the transfer of energy and the synthesis of several cellular components. Molecular and biochemical mechanisms related to how diatoms cope with P deficiency are not clear, and research into this has been limited to a few species. Among the molecular responses that have been reported in diatoms cultured under P deficient conditions is the upregulation of genes encoding enzymes related to the transport, assimilation, remobilization and recycling of this nutrient. Regarding biochemical responses, due to the reduction of the requirements for carbon structures for the synthesis of proteins and phospholipids, more CO2 is fixed than is consumed by the Calvin cycle. To deal with this excess, diatoms redirect the carbon flow toward the synthesis of storage compounds such as triacylglycerides and carbohydrates, which are excreted as extracellular polymeric substances. This review aimed to gather all current knowledge regarding the biochemical and molecular mechanisms of diatoms related to managing P deficiency in order to provide a wider insight into and understanding of their responses, as well as the metabolic pathways affected by the limitation of this nutrient.

Chili pepper (Capsicum spp.) is an important crop, as well as a model for fruit development studies and domestication. Here, we performed a time-course experiment to estimate standardized gene expression profiles with respect to fruit development for six domesticated and four wild chili pepper ancestors. We sampled the transcriptomes every 10 days from flowering to fruit maturity, and found that the mean standardized expression profiles for domesticated and wild accessions significantly differed. The mean standardized expression was higher and peaked earlier for domesticated vs. wild genotypes, particularly for genes involved in the cell cycle that ultimately control fruit size. We postulate that these gene expression changes are driven by selection pressures during domestication and show a robust network of cell cycle genes with a time shift in expression, which explains some of the differences between domesticated and wild phenotypes.

Jatropha species have been shown to be an important source of secondary metabolites with different biological effects. Jatropha cinerea (Ortega) Müll. Arg and Jatropha cordata (Ortega) Müll. Arg are distributed in the Northwestern region of Mexico, are adapted to extreme weather conditions and are widely used (stems, leaves, and sap) in traditional medicine. The aim of the present study was to carry out the phytochemical characterization and the evaluation of the antioxidant activity in methanolic extracts of stems and leaves from J. cinerea and J. cordata. The compounds present in the extracts of both species were characterized by ESI-IT-MS/MS and quantified by HPLC-DAD. The results showed that the stem extracts of both species are rich in phenolic acids, while the leaf extracts are rich in flavonoids. Some of the main compounds found were gallic acid, gentisic acid, 3,4-Dihydroxybenzoic acid, vitexin, isovitexin, and catechol. Both species showed high concentrations of phenols and total flavonoids and antioxidant activity. J. cordata showed the highest antioxidant capacity and the highest concentration of phenolic compounds. Overall, both Jatropha species are a natural source of antioxidant compounds with potential biotechnological uses.

ABSTRACT: Plants belonging to genus Jatropha has arisen interest because of its high oil content that could be used to produce biodiesel. It is also widely reported that the main fatty acids in Jatropha oilseed are oleic and linoleic acids. However, there are scarce studies related to native species of Jatropha from Northwestern Mexico which are adapted to arid conditions, and the expression of genes involved in fatty acid synthesis for these species is still unknown. Therefore, the aim of this study was to analyze the expression of five genes, ACP1, KASII, D9SD, FAD2-1 and FAD2-2, which are involved in the oleic and linoleic acids synthesis in mature wild-seeds of Jatropha cinerea, a native species from Sonoran Desert, using semi-quantitative RT-PCR. The J. cinerea seeds were randomly collected in Bahía de Kino, Sonora (México) which is a region characterized by its harsh environments such as saline soils and extreme temperature changes and J. curcas mature seeds from a non-toxic variety from Veracruz, Mexico were used as a reference. The RT-PCR analysis of three biological replicates were considered to ensure data consistent. Our analysis showed a higher expression of KASII and FAD2-1 genes in J. cinerea seeds compared to J. curcas, meanwhile the expression of ACP1, D9SD and FAD2-2 were higher in J. curcas. Furthermore, Actin and FAD2-1 genes sequences here obtained are the first reported for J. cinerea, thus providing information to develop further studies.

The strategy of bulk DNA sampling has been a valuable method for studying large numbers of individuals through genetic markers. The application of this strategy for discrimination among germplasm sources was analyzed through information theory, considering the case of polymorphic alleles scored binarily for their presence or absence in DNA pools. We defined the informativeness of a set of marker loci in bulks as the mutual information between genotype and population identity, composed by two terms: diversity and noise. The first term is the entropy of bulk genotypes, whereas the noise term is measured through the conditional entropy of bulk genotypes given germplasm sources. Thus, optimizing marker information implies increasing diversity and reducing noise. Simple formulas were devised to estimate marker information per allele from a set of estimated allele frequencies across populations. As an example, they allowed optimization of bulk size for SSR genotyping in maize, from allele frequencies estimated in a sample of 56 maize populations. It was found that a sample of 30 plants from a random mating population is adequate for maize germplasm SSR characterization. We analyzed the use of divided bulks to overcome the allele dilution problem in DNA pools, and concluded that samples of 30 plants divided into three bulks of 10 plants are efficient to characterize maize germplasm sources through SSR with a good control of the dilution problem. We estimated the informativeness of 30 SSR loci from the estimated allele frequencies in maize populations, and found a wide variation of marker informativeness, which positively correlated with the number of alleles per locus.

BACKGROUND: Drought is one of the major constraints for plant productivity worldwide. Different mechanisms of drought-tolerance have been reported for several plant species including maize. However, the differences in global gene expression between drought-tolerant and susceptible genotypes and their relationship to physiological adaptations to drought are largely unknown. The study of the differences in global gene expression between tolerant and susceptible genotypes could provide important information to design more efficient breeding programs to produce maize varieties better adapted to water limiting conditions. METHODOLOGY/PRINCIPAL FINDINGS: Changes in physiological responses and gene expression patterns were studied under drought stress and recovery in three Mexican maize landraces which included two drought tolerant (Cajete criollo and Michoacán 21) and one susceptible (85-2) genotypes. Photosynthesis, stomatal conductance, soil and leaf water potentials were monitored throughout the experiment and microarray analysis was carried out on transcripts obtained at 10 and 17 days following application of stress and after recovery irrigation. The two tolerant genotypes show more drastic changes in global gene expression which correlate with different physiological mechanisms of adaptation to drought. Differences in the kinetics and number of up- and down-regulated genes were observed between the tolerant and susceptible maize genotypes, as well as differences between the two tolerant genotypes. Interestingly, the most dramatic differences between the tolerant and susceptible genotypes were observed during recovery irrigation, suggesting that the tolerant genotypes activate mechanisms that allow more efficient recovery after a severe drought. CONCLUSIONS/SIGNIFICANCE: A correlation between levels of photosynthesis and transcription under stress was observed and differences in the number, type and expression levels of transcription factor families were also identified under drought and recovery between the three maize landraces. Gene expression analysis suggests that the drought tolerant landraces have a greater capacity to rapidly modulate more genes under drought and recovery in comparison to the susceptible landrace. Modulation of a greater number of differentially expressed genes of different TF gene families is an important characteristic of the tolerant genotypes. Finally, important differences were also noted between the tolerant landraces that underlie different mechanisms of achieving tolerance.

Las plantas del género Jatropha, pertenecientes a la familia de las Euphorbiaceae, pueden ser consideradas como alternativa para la obtención de semilla, con uso potencial en la fabricación de biocombustibles por su contenido alto de aceite. Se conocen alrededor de 186 especies distribuidas alrededor del mundo, de las cuales 48 se encuentran en México. En Sonora se conocen principalmente las especies de Jatropha cardiophylla, Jatropha cordata, Jatropha cinerea y Jatropha cuneata. La información que se tiene hasta el momento para especies adaptadas a zonas áridas en cuanto a la composición química de sus semillas es escasa y se desconoce el aspecto molecular relacionado con la síntesis de ácidos grasos. Por tal razón, se ha iniciado el estudio de la composición química y su relación con la expresión de genes, involucrados en la síntesis de ácidos grasos específicamente de los ácidos oleico y linoleico en las semillas de las especies de Jatropha nativas de Sonora.

As maize was domesticated in Mexico around 9,000 years ago, local farmers have selected and maintained seed stocks with particular traits and adapted to local conditions. In the present day, many of these landraces are still cultivated; however, increased urbanization and migration from rural areas implies a risk that this invaluable maize germplasm may be lost. In order to implement an efficient mechanism of conservation in situ, the diversity of these landrace populations must be estimated. Development of a method to select the minimum number of samples that would include the maximum number of alleles and identify germplasm harboring rare combinations of particular alleles will also safeguard the efficient ex-situ conservation of this germplasm. To reach this goal, a strategy based on SSR analysis and a novel algorithm to define a minimum collection and rare genotypes using landrace populations from Puebla State, Mexico, was developed as a "proof of concept" for methodology that could be extended to all maize landrace populations in Mexico and eventually to other native crops. The SSR-based strategy using bulked DNA samples allows rapid processing of large numbers of samples and can be set up in most laboratories equipped for basic molecular biology. Therefore, continuous monitoring of landrace populations locally could easily be carried out. This methodology can now be applied to support incentives for small farmers for the in situ conservation of these traditional cultivars.