Biofuel Crop Species Research Articles

High SNP diversity in the non-toxic indigenous Jatropha curcas germplasm widens the potential of this upcoming major biofuel crop species.

Jatropha curcas (jatropha) is an oil crop cultivated in (sub)tropical regions around the world, and holds great promise as a renewable energy source. However, efforts to fully commercialize jatropha are currently hampered by the lack of genetic diversity in the extant breeding germplasm, and by the toxicity of its seeds meaning that its seed cake cannot be used as a protein source in animal feed, among other constraints. In Mexico, the species' native range, there are jatropha plants whose seeds are used to prepare traditional meals. This non-toxic jatropha 'type' is considered to harbour low genetic variation due to a presumed domestication bottleneck and therefore to be of limited breeding value; yet, very little is known regarding its origin and genetic diversity. Using genotyping-by-sequencing (GBS), we extensively genotyped both indigenous toxic and non-toxic jatropha collected along roads and home gardens throughout southern Mexico. Single nucleotide polymorphism diversity in non-toxic jatropha is relatively high, particularly in northern Veracruz state, the probable origin of this germplasm. Genetic differences between toxic and non-toxic indigenous genotypes are overall quite small. A a genome-wide association study supported a genomic region (on LG 8, scaffold NW_012130064), probably involved in the suppression of seed toxicity. Conservation actions are urgently needed to preserve this non-toxic indigenous, relatively wild germplasm, having potential as a fuel feedstock, animal feed and food source among other uses. More generally, this work demonstrates the value of conservation genomic research on the indigenous gene pool of economically important plant species.

Characterization of drought- and heat-responsive microRNAs in switchgrass

Recent investigations revealed that microRNAs (miRNAs) play crucial roles in plant acclimation to stress conditions. Switchgrass, one of the important biofuel crop species can withstand hot and dry climates but the molecular basis of stress tolerance is relatively unknown. To identify miRNAs that are important for tolerating drought or heat, small RNAs were profiled in leaves of adult plants exposed to drought or heat. Sequence analysis enabled the identification of 29 conserved and 62 novel miRNA families. Notably, the abundances of several conserved and novel miRNAs were dramatically altered following drought or heat. Using at least one fold (log2) change as cut off, we observed that 13 conserved miRNA families were differentially regulated by both stresses, and, five and four families were specifically regulated by drought and heat, respectively. Similarly, using a more stringent cut off of two fold (log2) regulation, we found 5 and 16 novel miRNA families were upregulated but 6 and 7 families were downregulated under drought and heat, respectively. The stress-altered expression of a subset of miRNAs and their targets was confirmed using quantitative PCR. Overall, the switchgrass plants exposed to drought or heat revealed similarities as well as differences with respect to miRNA regulation, which could be important for enduring different stress conditions.

Photosynthetic energy conversion efficiency: setting a baseline for gauging future improvements in important food and biofuel crops.

The conversion efficiency (ε(c)) of absorbed radiation into biomass (MJ of dry matter per MJ of absorbed photosynthetically active radiation) is a component of yield potential that has been estimated at less than half the theoretical maximum. Various strategies have been proposed to improve ε(c), but a statistical analysis to establish baseline ε(c) levels across different crop functional types is lacking. Data from 164 published ε(c) studies conducted in relatively unstressed growth conditions were used to determine the means, greatest contributors to variation, and genetic trends in ε(c )across important food and biofuel crop species. ε(c) was greatest in biofuel crops (0.049-0.066), followed by C4 food crops (0.046-0.049), C3 nonlegumes (0.036-0.041), and finally C3 legumes (0.028-0.035). Despite confining our analysis to relatively unstressed growth conditions, total incident solar radiation and average growing season temperature most often accounted for the largest portion of ε(c) variability. Genetic improvements in ε(c), when present, were less than 0.7% per year, revealing the unrealized potential of improving ε(c) as a promising contributing strategy to meet projected future agricultural demand.

SNP Discovery with EST and NextGen Sequencing in Switchgrass (Panicum virgatum L.)

Although yield trials for switchgrass (Panicum virgatum L.), a potentially high value biofuel feedstock crop, are currently underway throughout North America, the genetic tools for crop improvement in this species are still in the early stages of development. Identification of high-density molecular markers, such as single nucleotide polymorphisms (SNPs), that are amenable to high-throughput genotyping approaches, is the first step in a quantitative genetics study of this model biofuel crop species. We generated and sequenced expressed sequence tag (EST) libraries from thirteen diverse switchgrass cultivars representing both upland and lowland ecotypes, as well as tetraploid and octoploid genomes. We followed this with reduced genomic library preparation and massively parallel sequencing of the same samples using the Illumina Genome Analyzer technology platform. EST libraries were used to generate unigene clusters and establish a gene-space reference sequence, thus providing a framework for assembly of the short sequence reads. SNPs were identified utilizing these scaffolds. We used a custom software program for alignment and SNP detection and identified over 149,000 SNPs across the 13 short-read sequencing libraries (SRSLs). Approximately 25,000 additional SNPs were identified from the entire EST collection available for the species. This sequencing effort generated data that are suitable for marker development and for estimation of population genetic parameters, such as nucleotide diversity and linkage disequilibrium. Based on these data, we assessed the feasibility of genome wide association mapping and genomic selection applications in switchgrass. Overall, the SNP markers discovered in this study will help facilitate quantitative genetics experiments and greatly enhance breeding efforts that target improvement of key biofuel traits and development of new switchgrass cultivars.

Bioenergy Crops Miscanthus × giganteus and Panicum virgatum Reduce Growth and Survivorship of Spodoptera frugiperda (Lepidoptera: Noctuidae)

Large-scale cultivation of plants used as biofuels is likely to alter the ecological interactions of current agricultural crops and their insect pests in a myriad of ways. Recent evidence suggests many contemporary maize pests will be able to use potential biofuel crops such as switchgrass, Panicum virgatum L., and miscanthus as hosts. To determine how suitable these biofuels are to the maize, Zea mays L., pest and generalist graminivore, Spodoptera frugiperda (J. E. Smith) (Lepidoptera: Noctuidae), we examined host plant preference and larval performance on foliage grown for commercial biofuel production. Larvae fed leaf tissue from both field- and greenhouse-grown switchgrass and miscanthus were monitored for survival, development, and food use relative to field-grown maize. Survivorship on biofuel crops was high on greenhouse-grown leaf tissue but severely reduced for field-grown switchgrass, and no larvae survived on field-grown miscanthus. Larvae fed field-grown tissue had larger head capsules yet achieved lower pupal weights because the increased toughness of the leaf tissue prevented the assimilation of nitrogen. Given that larvae overwhelmingly preferred maize to other biofuel crop species and that survival and performance were dramatically reduced on biofuel crop species, it is likely that biofuel crops, as grown for field cultivation, will suffer reduced damage from maize pests such as S. frugiperda because of reduced suitability.