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Microthlaspi erraticum is widely distributed in temperate Eurasia, but restricted to Ca2+-rich habitats, predominantly on white Jurassic limestone, which is made up by calcium carbonate, with little other minerals. Thus, naturally occurring Microthlaspi erraticum individuals are confronted with a high concentration of Ca2+ ions while Mg2+ ion concentration is relatively low. As there is a competitive uptake between these two ions, adaptation to the soil condition can be expected. In this study, it was the aim to explore the genomic consequences of this adaptation by sequencing and analysing the genome of Microthlaspi erraticum . Its genome size is comparable with other diploid Brassicaceae, while more genes were predicted. Two Mg2+ transporters known to be expressed in roots were duplicated and one showed a significant degree of positive selection. It is speculated that this evolved due to the pressure to take up Mg2+ ions efficiently in the presence of an overwhelming amount of Ca2+ ions. Future studies on plants specialized on similar soils and affinity tests of the transporters are needed to provide unequivocal evidence for this hypothesis. If verified, the transporters found in this study might be useful for breeding Brassicaceae crops for higher yield on Ca2+-rich and Mg2+ -poor soils.

Summary Thlaspi arvense (field pennycress) is being domesticated as a winter annual oilseed crop capable of improving ecosystems and intensifying agricultural productivity without increasing land use. It is a selfing diploid with a short life cycle and is amenable to genetic manipulations, making it an accessible field‐based model species for genetics and epigenetics. The availability of a high‐quality reference genome is vital for understanding pennycress physiology and for clarifying its evolutionary history within the Brassicaceae. Here, we present a chromosome‐level genome assembly of var. MN106‐Ref with improved gene annotation and use it to investigate gene structure differences between two accessions (MN108 and Spring32‐10) that are highly amenable to genetic transformation. We describe non‐coding RNAs, pseudogenes and transposable elements, and highlight tissue‐specific expression and methylation patterns. Resequencing of forty wild accessions provided insights into genome‐wide genetic variation, and QTL regions were identified for a seedling colour phenotype. Altogether, these data will serve as a tool for pennycress improvement in general and for translational research across the Brassicaceae.

Summary Thlapsi arvense L. (pennycress) is being developed as a profitable oilseed cover crop for the winter fallow period throughout the temperate regions of the world, controlling soil erosion and nutrients run‐off on otherwise barren farmland. We demonstrate that pennycress can serve as a user‐friendly model system akin to Arabidopsis that is well‐suited for both laboratory and field experimentation. We sequenced the diploid genome of the spring‐type Spring 32‐10 inbred line (1C DNA content of 539 Mb; 2n = 14), identifying variation that may explain phenotypic differences with winter‐type pennycress, as well as predominantly a one‐to‐one correspondence with Arabidopsis genes, which makes translational research straightforward. We developed an Agrobacterium‐mediated floral dip transformation method (0.5% transformation efficiency) and introduced CRISPR‐Cas9 constructs to produce indel mutations in the putative FATTY ACID ELONGATION1 (FAE1) gene, thereby abolishing erucic acid production and creating an edible seed oil comparable to that of canola. We also stably transformed pennycress with the Euonymus alatus diacylglycerol acetyltransferase (EaDAcT) gene, producing low‐viscosity acetyl‐triacylglycerol‐containing seed oil suitable as a diesel‐engine drop‐in fuel. Adoption of pennycress as a model system will accelerate oilseed‐crop translational research and facilitate pennycress’ rapid domestication to meet the growing sustainable food and fuel demands.

There is increasing knowledge on the diversity of root-endophytic fungi, but limited information on their lifestyles and dependence on hosts hampers our understanding of their ecological functions. We compared diversity and biogeographical patterns of cultivable and noncultivable root endophytes to assess whether their occurrence is determined by distinct ecological factors. The endophytic diversity in roots of nonmycorrhizal Microthlaspi spp. growing across Europe was assessed using high-throughput sequencing (HTS) and compared with a previous dataset based on cultivation of endophytes from the same root samples. HTS revealed a large fungal richness undetected by cultivation, but which largely comprised taxa with restricted distributions and/or low representation of sequence reads. Both datasets coincided in a consistent high representation of widespread endophytes within orders Pleosporales, Hypocreales and Helotiales, as well as similar associations of community structure with spatial and environmental conditions. Likewise, distributions of particular endophytes inferred by HTS agreed with cultivation data in suggesting individual ecological preferences. Our findings support that Microthlaspi spp. roots are colonized mostly by saprotrophic and likely facultative endophytes, and that differential niche preferences and distribution ranges among fungi importantly drive the assembly of root-endophytic communities.

Pennycress ( Thlaspi arvense L.) is being domesticated as an oilseed cash cover crop to be grown in the off-season throughout temperate regions of the world. With its diploid genome and ease of directed mutagenesis using molecular approaches, pennycress seed oil composition can be rapidly tailored for a plethora of food, feed, oleochemical and fuel uses. Here, we utilized Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR)/Cas9 technology to produce knockout mutations in the FATTY ACID DESATURASE2 ( FAD2 ) and REDUCED OLEATE DESATURATION1 ( ROD1 ) genes to increase oleic acid content. High oleic acid (18:1) oil is valued for its oxidative stability that is superior to the polyunsaturated fatty acids (PUFAs) linoleic (18:2) and linolenic (18:3), and better cold flow properties than the very long chain fatty acid (VLCFA) erucic (22:1). When combined with a FATTY ACID ELONGATION1 ( fae1 ) knockout mutation, fad2 fae1 and rod1 fae1 double mutants produced ∼90% and ∼60% oleic acid in seed oil, respectively, with PUFAs in fad2 fae1 as well as fad2 single mutants reduced to less than 5%. MALDI-MS spatial imaging analyses of phosphatidylcholine (PC) and triacylglycerol (TAG) molecular species in wild-type pennycress embryo sections from mature seeds revealed that erucic acid is highly enriched in cotyledons which serve as storage organs, suggestive of a role in providing energy for the germinating seedling. In contrast, PUFA-containing TAGs are enriched in the embryonic axis, which may be utilized for cellular membrane expansion during seed germination and seedling emergence. Under standard growth chamber conditions, rod1 fae1 plants grew like wild type whereas fad2 single and fad2 fae1 double mutant plants exhibited delayed growth and overall reduced heights and seed yields, suggesting that reducing PUFAs below a threshold in pennycress had negative physiological effects. Taken together, our results suggest that combinatorial knockout of ROD1 and FAE1 may be a viable route to commercially increase oleic acid content in pennycress seed oil whereas mutations in FAD2 will likely require at least partial function to avoid fitness trade-offs.

Background:Biomass‑derived jet fuel is an alternative jet fuel (AJF) showing promise of reducing the dependence on fossil fuel and greenhouse gas emissions. Hydroprocessed esters and fatty acids (HEFA) concept is also known as one of the pathways for producing bio jet fuel. HEFA fuel was approved by the American Society for Testing and Materials in 2011, and can be blended up to 50% with conventional jet fuel. Since then, several HEFA economic and life‑cycle assessments have been published in literature. However, there have been limited analyses on feedstock availability,composition, and their impact on hydrocarbon yield (particularly jet blendstock yield) and overall process economics.Results:This study examines over 20 oil feedstocks, their geographic distribution and production levels, oil yield, prices, and chemical composition. The results of our compositional analysis indicate that most oils contain mainly C16 and C18 fatty acids except pennycress, yellow grease, and mustard, which contain higher values and thus would require hydrocracking to improve jet fuel production. Coconut oil has a large content of shorter carbon fatty acids, making it a good feedstock candidate for renewable gasoline instead of jet substitutes’ production. Techno‑economic analysis (TEA) was performed for five selected oil feedstocks—camelina, pennycress, jatropha, castor bean, and yellow grease—using the HEFA process concept.Conclusion:The resource analysis indicates that oil crops currently grown in the United States (namely soybean) have relatively low oil yield when compared to oil crops grown in other parts of the world, such as palm, coconut, and jatropha. Also, non‑terrestrial oil sources, such as animal fats and greases, have relatively lower prices than terrestrial oil crops. The minimum jet fuel selling price for these five resources ranges between$3.8 and $ 11.0 per gallon. The results of our TEA and resource studies indicate the key cost drivers for a biorefinery converting oil to jet hydrocarbons are as follows: oil price, conversion plant capacity, fatty acid profile, addition of hydrocracker, and type of hydroprocessing catalysts.

The lipidome comprises the total content of molecular species of each lipid class, and is measured using the analytical techniques of lipidomics. Many liquid chromatography-mass spectrometry (LC-MS) methods have previously been described to characterize the lipidome. However, many lipidomic approaches may not fully uncover the subtleties of lipid molecular species, such as the full fatty acid (FA) composition of certain lipid classes. Here, we describe a stepwise targeted lipidomics approach to characterize the polar and non-polar lipid classes using complementary LC-MS methods. Our “polar” method measures 260 molecular species across 12 polar lipid classes, and is performed using hydrophilic interaction chromatography (HILIC) on a NH2 column to separate lipid classes by their headgroup. Our “non-polar” method measures 254 molecular species across three non-polar lipid classes, separating molecular species on their FA characteristics by reverse phase (RP) chromatography on a C30 column. Five different extraction methods were compared, with an MTBE-based extraction chosen for the final lipidomics workflow. A state-of-the-art strategy to determine and relatively quantify the FA composition of triacylglycerols is also described. This lipidomics workflow was applied to developing, mature, and germinated pennycress seeds/seedlings and found unexpected changes among several lipid molecular species. During development, diacylglycerols predominantly contained long chain length FAs, which contrasted with the very long chain FAs of triacylglycerols in mature seeds. Potential metabolic explanations are discussed. The lack of very long chain fatty acids in diacylglycerols of germinating seeds may indicate very long chain FAs, such as erucic acid, are preferentially channeled into beta-oxidation for energy production.

Aims This study aimed at revealing the mechanisms of zinc(Zn)-induced root growth inhibition in the Zn hyperaccumulator Noccaea caerulescens and the excluder Microthlaspi perfoliatum to shed light on the intriguing question whether there is any selectivity in the Zn effects on root cell division and elongation. Methods Zinc effects on various parameters characterizing root cell division and elongation were studied. Total Zn uptake, accumulation, root-to-shoot translocation, and distribution over the root tip tissues as well as metal-induced oxidative stress were also assessed. Results Similar degrees of root growth inhibition were achieved when the Zn concentration in the medium and Zn content in the roots were 150 and 5 times higher, respectively, in the hyperaccumulator, compared to the excluder. Zinc accumulated in the cell walls and protoplasts in the root meristem and elongation zone. Although Zn negatively affected both root cell division and elongation, at a similar degree of root growth inhibition, the contribution of the inhibition of cell division was greater in N. caerulescens compared to M. perfoliatum , as the decrease in the meristem length, the number of meristematic cells in a file, and the mitotic index were more prominent in the hyperaccumulator. Conclusions Greater contribution of the inhibition of cell division in N. caerulescens compared to M. perfoliatum at a similar degree of root growth inhibition seemed to be partly determined by a stronger degree of oxidative stress and disturbance of mineral nutrition in the roots of the hyperaccumulator, owing to a much higher Zn accumulation.

Main ConclusionPennycress, as an emerging oilseed crop with high oil content, faces challenges but offers potential for sustainable bioproducts; ongoing research aims to enhance its traits and quality.Pennycress (Thlaspi arvense L.) is an emerging oilseed crop with many advantages, such as high seed oil (27–39%) and monounsaturated fatty acid (55.6%) content, making it an attractive candidate to produce sustainable bioproducts. However, several challenges are associated with domesticating pennycress, including high silicle shatter, which reduces seed yield during harvest, non-uniformed germination rate and high contents of erucic acid and glucosinolates, which have adverse health effects on humans and animals. Pennycress, which can be easily and rapidly transformed using the floral dip method under vacuum, can achieve trait improvements. Ongoing research for pennycress domestication using mutation breeding, including ethylmethylsulfonate treatment and genome editing, aims to improve its quality. Pennycress can be used as an excellent platform for producing industrially important fatty acids such as hydroxy and epoxy fatty acids and docosahexaenoic acid. In conclusion, pennycress is a promising oilseed crop with multiple advantages and potential applications. Continuous improvements in quality and engineering for producing high-value bio-based feedstocks in pennycress will establish it as a sustainable and economically valuable crop.

Background Understanding how organisms evolve and adapt to extreme habitats is of crucial importance in evolutionary ecology. Altitude gradients are an important determinant of the distribution pattern and range of organisms due to distinct climate conditions at different altitudes. High-altitude regions often provide extreme environments including low temperature and oxygen concentration, poor soil, and strong levels of ultraviolet radiation, leading to very few plant species being able to populate elevation ranges greater than 4000 m. Field pennycress ( Thlaspi arvense ) is a valuable oilseed crop and emerging model plant distributed across an elevation range of nearly 4500 m. Here, we generate an improved genome assembly to understand how this species adapts to such different environments. Results We sequenced and assembled de novo the chromosome-level pennycress genome of 527.3 Mb encoding 31,596 genes. Phylogenomic analyses based on 2495 single-copy genes revealed that pennycress is closely related to Eutrema salsugineum (estimated divergence 14.32–18.58 Mya), and both species form a sister clade to Schrenkiella parvula and genus Brassica . Field pennycress contains the highest percentage (70.19%) of transposable elements in all reported genomes of Brassicaceae, with the retrotransposon proliferation in the Middle Pleistocene being likely responsible for the expansion of genome size. Moreover, our analysis of 40 field pennycress samples in two high- and two low-elevation populations detected 1,256,971 high-quality single nucleotide polymorphisms. Using three complementary selection tests, we detected 130 candidate naturally selected genes in the Qinghai-Tibet Plateau (QTP) populations, some of which are involved in DNA repair and the ubiquitin system and potential candidates involved in high-altitude adaptation. Notably, we detected a single base mutation causing loss-of-function of the FLOWERING LOCUS C protein, responsible for the transition to early flowering in high-elevation populations. Conclusions Our results provide a genome-wide perspective of how plants adapt to distinct environmental conditions across extreme elevation differences and the potential for further follow-up research with extensive data from additional populations and species.