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At least 91 records · Page 5

A Chromosome-Scale Genome Assembly of a Helicoverpa zea Strain Resistant to Bacillus thuringiensis Cry1Ac Insecticidal Protein

Helicoverpa zea (Lepidoptera: Noctuidae) is an insect pest of major cultivated crops in North and South America. The species has adapted to different host plants and developed resistance to several insecticidal agents, including Bacillus thuringiensis (Bt) insecticidal proteins in transgenic cotton and maize. Helicoverpa zea populations persist year-round in tropical and subtropical regions, but seasonal migrations into temperate zones increase the geographic range of associated crop damage. To better understand the genetic basis of these physiological and ecological characteristics, we generated a high-quality chromosome-level assembly for a single H. zea male from Bt-resistant strain, HzStark_Cry1AcR. Hi-C data were used to scaffold an initial 375.2 Mb contig assembly into 30 autosomes and the Z sex chromosome (scaffold N50 = 12.8 Mb and L50 = 14). The scaffolded assembly was error-corrected with a novel pipeline, polishCLR. The mitochondrial genome was assembled through an improved pipeline and annotated. Assessment of this genome assembly indicated 98.8% of the Lepidopteran Benchmark Universal Single-Copy Ortholog set were complete (98.5% as complete single copy). Repetitive elements comprised approximately 29.5% of the assembly with the plurality (11.2%) classified as retroelements. This chromosome-scale reference assembly for H. zea, ilHelZeax1.1, will facilitate future research to evaluate and enhance sustainable crop production practices.

59 BASIC BIOLOGICAL SCIENCES↗

Identification of Novel Genomic Associations and Gene Candidates for Grain Starch Content in Sorghum

Starch accumulated in the endosperm of cereal grains as reserve energy for germination serves as a staple in human and animal nutrition. Unraveling genetic control for starch metabolism is important for breeding grains with high starch content. In this study, we used a sorghum association panel with 389 individuals and 141,557 single nucleotide polymorphisms (SNPs) to fit linear mixed models (LMM) for identifying genomic regions and potential candidate genes associated with starch content. Three associated genomic regions, one in chromosome (chr) 1 and two novel associations in chr-8, were identified using combination of LMM and Bayesian sparse LMM. All significant SNPs were located within protein coding genes, with SNPs ∼ 52 Mb of chr-8 encoding a Casperian strip membrane protein (CASP)-like protein (Sobic.008G111500) and a heat shock protein (HSP) 90 (Sobic.008G111600) that were highly expressed in reproductive tissues including within the embryo and endosperm. The HSP90 is a potential hub gene with gene network of 75 high-confidence first interactors that is enriched for five biochemical pathways including protein processing. The first interactors of HSP90 also showed high transcript abundance in reproductive tissues. The candidates of this study are likely involved in intricate metabolic pathways and represent candidate gene targets for source-sink activities and drought and heat stress tolerance during grain filling.

genomics↗

Genetic dissection of natural variation in oilseed traits of camelina by whole‐genome resequencing and QTL mapping

Abstract Camelina [ Camelina sativa (L.) Crantz] is an oilseed crop in the Brassicaceae family that is currently being developed as a source of bioenergy and healthy fatty acids. To facilitate modern breeding efforts through marker‐assisted selection and biotechnology, we evaluated genetic variation among a worldwide collection of 222 camelina accessions. We performed whole‐genome resequencing to obtain single nucleotide polymorphism (SNP) markers and to analyze genomic diversity. We also conducted phenotypic field evaluations in two consecutive seasons for variations in key agronomic traits related to oilseed production such as seed size, oil content (OC), fatty acid composition, and flowering time. We determined the population structure of the camelina accessions using 161,301 SNPs. Further, we identified quantitative trait loci (QTL) and candidate genes controlling the above field‐evaluated traits by genome‐wide association studies (GWAS) complemented with linkage mapping using a recombinant inbred line (RIL) population. Characterization of the natural variation at the genome and phenotypic levels provides valuable resources to camelina genetic studies and crop improvement. The QTL and candidate genes should assist in breeding of advanced camelina varieties that can be integrated into the cropping systems for the production of high yield of oils of desired fatty acid composition.

59 BASIC BIOLOGICAL SCIENCES↗

3P Program: Phenotyping X Prediction = Productivity (Final Scientific/Technical Report)

The goal of the 3P Program was to establish integrated, real-time phenotyping and to analyze above- and below-ground plant architecture and total carbon partitioning and allocation to predict heterosis and develop superior crop hybrids by fully leveraging the Sorghum gene pool. There were two overarching themes: 1) the development of a new crop improvement approach utilizing advances in high-throughput phenotyping (HTP), computing, and genomics for public dissemination and 2) leveraging this platform for sorghum crop improvement and commercialization. The Clemson team worked on creating genomic resources and using both statistical learning and high-throughput phenotyping in genomics-assisted breeding. Research was broadly interested in the genetics of carbon partitioning, with the aim of improving crop performance and achieving sustainability. The technology and resources created can be readily found in the public domain and serve to advance scientific understanding of crop genomics and breeding. Genomic prediction was able to identify top crosses to be made, and a hybrid prediction pipeline is in place to drive year-over-year genetic gain. Roots have long been ignored by plant breeders and agronomists, not because they are unimportant but because they are hard to measure. This is an untapped white space of potential insight and innovation. To address this, Hi Fidelity Genetics developed the RootTracker to measure roots in the field on a continuous basis. A database system called RootTracker Tracker was developed to handle data coming from the RootTrackers. In using this device, valuable data was observed for plant breeding, hydrochemical development, and other agricultural biology applications. Carnegie Mellon’s goal was developing new techniques to generate high-resolution 3D models of plants from data collected in the field. The idea was that more useful and more informative phenotypes could be extracted by resolving small features, such as seeds and flowers, and that by modeling in 3D, the spatial structure of plants could be examined. To achieve this, multiple images collected by a new small format structured light stereo imager were fused together. A sorghum panicle modeling pipeline was developed to allow the collection and processing of data. Carolina Seed Systems is an agricultural technology company focused on decarbonizing the agricultural system. Their technology pipeline serves to drive fundamental progress towards creation and distribution of carbon negative crops. The genomic and the engineering technology developed through the 3P Program was leveraged to deliver both value and sustainability from the grower to the consumer. Promising sorghum hybrids were scaled up and commercialized. The overall goal of our research was to integrate, create, and deploy genetic and engineering concepts and technologies to enhance crop productivity in a sustainable fashion. The combination of public and private partners allowed the basic research and hypothesis testing to be quickly accelerated for commercial application by the companies yet maintained that the core framework and academic insights remain in the public domain for continued market disruption, competition, and innovation.

59 BASIC BIOLOGICAL SCIENCES↗

Chapter 6: An Improved Leaf Protoplast System for Highly Efficient Transient Expression in Switchgrass (Panicum virgatum L.)

As a robust perennial C4-type monocot plant and a native species to North America, switchgrass (Panicum virgatum) has been evaluated and designated as a strong candidate bioenergy crop by the U.S. DOE. Although genetic modifications of switchgrass have been used to successfully reduce the recalcitrance of switchgrass biomass for biofuel production, the generation of transgenic switchgrass is still a slow and laborious process. A transient protoplast system can provide an excellent platform to accelerate the selection of genes-of-interest for tailoring switchgrass biomass. However, partially due to the lack of the complete genomic information, the attempts to optimize the transient protoplast system for switchgrass remain scarce. In this chapter, we provide an improved protocol for switchgrass protoplast isolation, increased transformation efficiency using CsCl gradient ultracentrifugation-derived plasmid DNA and extended application of the transient switchgrass protoplast system to analyze protein expression using western blot.

BASIC BIOLOGICAL SCIENCES,BIOMASS FUELS↗

Marker-free carotenoid-enriched rice generated through targeted gene insertion using CRISPR-Cas9

Targeted insertion of transgenes at pre-determined plant genomic safe harbors provides a desirable alternative to insertions at random sites achieved through conventional methods. Most existing cases of targeted gene insertion in plants have either relied on the presence of a selectable marker gene in the insertion cassette or occurred at low frequency with relatively small DNA fragments (<1.8 kb). Here, we report the use of an optimized CRISPR-Cas9-based method to achieve the targeted insertion of a 5.2 kb carotenoid biosynthesis cassette at two genomic safe harbors in rice. We obtain marker-free rice plants with high carotenoid content in the seeds and no detectable penalty in morphology or yield. Whole-genome sequencing reveals the absence of off-target mutations by Cas9 in the engineered plants. These results demonstrate targeted gene insertion of marker-free DNA in rice using CRISPR-Cas9 genome editing, and offer a promising strategy for genetic improvement of rice and other crops.

59 BASIC BIOLOGICAL SCIENCES↗

Genetic variation in Zea mays influences microbial nitrification and denitrification in conventional agroecosystems

Nitrogenous fertilizers provide a short-lived benefit to crops in agroecosystems, but stimulate nitrification and denitrification, processes that result in nitrate pollution, N 2 O production, and reduced soil fertility. Recent advances in plant microbiome science suggest that genetic variation in plants can modulate the composition and activity of rhizosphere N-cycling microorganisms. Here we attempted to determine whether genetic variation exists in Zea mays for the ability to influence the rhizosphere nitrifier and denitrifier microbiome under “real-world” conventional agricultural conditions. To capture an extensive amount of genetic diversity within maize we grew and sampled the rhizosphere microbiome of a diversity panel of germplasm that included ex-PVP inbreds (Z. mays ssp. mays), ex-PVP hybrids (Z. mays ssp. mays), and teosinte (Z. mays ssp. mexicana and Z. mays ssp. parviglumis). From these samples, we characterized the microbiome, a suite of microbial genes involved in nitrification and denitrification and carried out N-cycling potential assays. Here we are showing that populations/genotypes of a single species can vary in their ecological interaction with denitrifers and nitrifers. Some hybrid and teosinte genotypes supported microbial communities with lower potential nitrification and potential denitrification activity in the rhizosphere, while inbred genotypes stimulated/did not inhibit these N-cycling activities. These potential differences translated to functional differences in N 2 O fluxes, with teosinte plots producing less GHG than maize plots. Taken together, these results suggest that Zea genetic variation can lead to changes in N-cycling processes that result in N leaching and N 2 O production, and thereby are selectable targets for crop improvement. Understanding the underlying genetic variation contributing to belowground microbiome N-cycling into our conventional agricultural system could be useful for sustainability.

59 BASIC BIOLOGICAL SCIENCES↗

Surveying Grassland Islands: the genetics and performance of Appalachian switchgrass (Panicum virgatum L.) collections

The interior Southeastern United States could contain novel germplasm for the bioenergy crop switchgrass due to its diverse habitats and geographic location between genetic subpopulations (Atlantic, Midwest, and Gulf). Collections from this region could accelerate breeding progress, contribute to conservation efforts, and improve understanding of isolated grasslands in the region. This study located 22 sites in the Midsouth region and obtained 1,521,210 single nucleotide polymorphism markers of 202 individuals through genotype-by-sequencing. Here, individuals were evaluated for flowering time, winter survival and tiller number. Comparison to a national diversity panel revealed that branches of two major subpopulations occur in the region with two levels of polyploidy: Atlantic tetraploids and Midwest octoploids. Two locations contained admixed octoploid individuals with Midwest and Gulf genetics. Field performance of the Midwest octoploids conformed with prior reported performance of the Midwest subpopulation, although three sites contained promising late flowering traits. The Atlantic tetraploids had moderate winter survival, short stature, and anomalously early flowering. Atlantic populations mostly occurred in marginal sites and their morphological and flowering time adaptations may be a resource conservation strategy. Demographic inference of historical effective population size variation in a subset of tetraploid locations indicated a widespread recent decline in effective population size. This pattern is consistent with isolation of these switchgrass communities from larger populations and is further supported by evidence of inbreeding within the populations (F I = 0.18). The populations documented in this study contain novel genetic diversity and adaptations to a range of marginal habitats. Therefore, this study provides a new source of germplasm for future breeding and conservation programs.

59 BASIC BIOLOGICAL SCIENCES↗

XERICO as a target for engineering stress-resilient crops: Mechanisms, applications, and future directions

XERICO's capacity to enhance ABA-driven stress responses across diverse crops, its regulatory crosstalk with other hormonal pathways, and its compatibility with advanced genetic engineering tools highlight its central role in sustainable agriculture. Leveraging XERICO in crop improvement programs aligns with the urgent need to mitigate the impacts of climate-induced stress in agriculture, offering a pathway toward resilient and high-yielding crops. Here, by enabling crops to withstand drought and other environmental stresses, XERICO-based biotechnological approaches hold transformative potential for global food security and environmental sustainability.

09 BIOMASS FUELS↗

Phenome‐to‐genome insights for evaluating root system architecture in field studies of maize

Abstract Understanding the genetic basis of root system architecture (RSA) in crops requires innovative approaches that enable both high‐throughput and precise phenotyping in field conditions. In this study, we evaluated multiple phenotyping and analytical frameworks for quantifying RSA in mature, field‐grown maize in three field experiments. We used forward and reverse genetic approaches to evaluate >1700 maize root crowns, including a diversity panel, a biparental mapping population, and maize mutant and wild‐type alleles at two known RSA genes,DEEPER ROOTING 1(DRO1) andRootless1(Rt1). We show the utility of increasing the dimensionality of traditional two‐dimensional (2D) techniques, referred to as the “2D multi‐view” method, to improve the capture of whole root system information for mapping genetic variation influencing RSA. Comparison of univariate and multivariate genome‐wide association study (GWAS) approaches revealed that multivariate traits were effective at dissecting complex RSA phenotypes and identifying pleiotropic quantitative trait loci (QTLs). Overall, three‐dimensional (3D) root models generated from X‐ray computed tomography and digital phenotyping captured a larger proportion of RSA trait variations compared to other methods of root phenotyping, as evidenced by both genome‐wide and single‐gene analyses. Among the individual root traits, root pulling force emerged as a highly heritable estimate of RSA that identified the largest number of shared QTLs with 3D phenotypes. Our study shows that integrating complementary phenotyping technologies helps to provide a more comprehensive understanding of the genetic architecture of RSA in field‐grown maize.

Genetics & Heredity↗

Biodiesel from oil produced in vegetative tissues of biomass – A review

Biodiesel is a green, renewable alternative to petroleum-derived diesel. However, using vegetable oil for bio- diesel production significantly challenges the food security. Progress in metabolic engineering, understanding of lipid biosynthesis and storage have enabled engineering of vegetative tissues of plants such as sugarcane, sor- ghum, and tobacco for lipid production. Such sources could be cultivated on land resources, which are currently not suitable for row crops. Besides achieving significant lipid accumulation, it is imperative to maintain the fatty acid and lipid profile ideal for biodiesel production and engine performance. In this study, genetic modifications used to induce lipid accumulation in transgenic crops and the proposed strategies for efficient recovery of oil from these crops have been presented. Here, this paper highlights that lipids sourced from vegetative biomass in their native form would pose significant challenges in biodiesel production. Therefore, different strategies have been presented for improving feedstock quality to achieve high-quality biodiesel production.

09 BIOMASS FUELS↗

Bioenergy crops as alternative feedstocks for recovery of anthocyanins: A review

Anthocyanins are plant-based pigments known for their wide range of industrial applications. Berries, grapes, purple yam, purple corn, and black rice are among the chief dietary sources of anthocyanins. However, these sources have a well-established market in the food sector and cannot be diverted completely for the extraction of anthocyanins. Hence, there is a need to identify alternative sources of these natural colorants. In this review, color-bearing bioenergy crops with high productivity such as miscanthus, sorghum, and sugarcane have been proposed as potential feedstocks for the recovery of anthocyanins as co-products. The role of genetic modification in the biosynthesis of anthocyanins in these crops has been discussed. A consolidated bioprocessing approach has been proposed for the recovery of anthocyanins from bioenergy crops along with the production of bioethanol and lignin-based chemicals. Such an integrated process could make the overall process for the production of biofuels cost-effective.

09 BIOMASS FUELS↗

Optimizing the electron transport chain to sustainably improve photosynthesis

Abstract Genetically improving photosynthesis is a key strategy to boosting crop production to meet the rising demand for food and fuel by a rapidly growing global population in a warming climate. Many components of the photosynthetic apparatus have been targeted for genetic modification for improving photosynthesis. Successful translation of these modifications into increased plant productivity in fluctuating environments will depend on whether the electron transport chain (ETC) can support the increased electron transport rate without risking overreduction and photodamage. At present atmospheric conditions, the ETC appears suboptimal and will likely need to be modified to support proposed photosynthetic improvements and to maintain energy balance. Here, I derive photochemical equations to quantify the transport capacity and the corresponding reduction level based on the kinetics of redox reactions along the ETC. Using these theoretical equations and measurements from diverse C3/C4 species across environments, I identify several strategies that can simultaneously increase the transport capacity and decrease the reduction level of the ETC. These strategies include increasing the abundances of reaction centers, cytochrome b6f complexes, and mobile electron carriers, improving their redox kinetics, and decreasing the fraction of secondary quinone–nonreducing photosystem II reaction centers. I also shed light on several previously unexplained experimental findings regarding the physiological impacts of the abundances of the cytochrome b6f complex and plastoquinone. The model developed, and the insights generated from it facilitate the development of sustainable photosynthetic systems for greater crop yields.

59 BASIC BIOLOGICAL SCIENCES↗

Growth and biochemical composition of the carrageenophyte Eucheumatopsis isiformis (Solieriaceae, Rhodophyta) under cultivation at different irradiance levels

Long-term production of a few species of Eucheuma and Kappaphycus from the Indo-Pacific Ocean supports the global carrageenan industry. The widespread use of vegetative propagation has resulted in low genetic diversity among farmed populations, making these crops increasingly vulnerable to diseases and environmental stressors, and resulting in declining yields and reduced biomass production over time. Insufficient efforts to develop and select new native cultivars, combined with the ecological risks posed by introducing nonindigenous species, underscore the urgent need to enhance cultivar diversification in eucheumatoid farming. Eucheumatopsis isiformis , a native carrageenophyte to the Atlantic Ocean, has potential for its cultivation in the Yucatán peninsula, Mexico. This study evaluated the growth and biochemical composition (pigments, elemental C and N, kappa/iota carrageenan ratio) of E. isiformis under different irradiance conditions in an indoor cultivation system. Specimens were collected in a subtidal population in Yucatán, Mexico, characterized, acclimatized to laboratory conditions, and exposed to five irradiance levels (71 ± 10.59 –1014 ± 65.85 µmol quanta m −2 s −1 ; 23.4 °C, salinity 33–34). Growth was measured weekly for six weeks, and elemental C and N content, pigment composition, and the kappa/iota-carrageenan ratio were evaluated. Growth and chromatic photoacclimation to light conditions were observed in all treatments. Highest growth rates (3.1–3.2% day −1 ) were registered between 238 ± 28.3 – 518 ± 44.3 µmol quanta m −2 s −1 , with a predominant iota-type carrageenan (0.97 ± 0.02) observed in all treatments. This study aims to provide guidelines for the successful development of the aquaculture of this carrageenophyte.

FTIR↗

Patch selection by bumble bees navigating discontinuous landscapes

Pollen and nectar resources are unevenly distributed over space and bees must make routing decisions when navigating patchy resources. Determining the patch selection process used by bees is crucial to understanding bee foraging over discontinuous landscapes. To elucidate this process, we developed four distinct probability models of bee movement where the size and the distance to the patch determined the attractiveness of a patch. A field experiment with a center patch and four peripheral patches of two distinct sizes and distances from the center was set up in two configurations. Empirical transition probabilities from the center to each peripheral patch were obtained at two sites and two years. The best model was identified by comparing observed and predicted transition probabilities, where predicted values were obtained by incorporating the spatial dimensions of the field experiment into each model’s mathematical expression. Bumble bees used both patch size and isolation distance when selecting a patch and could assess the total amount of resources available in a patch. Bumble bees prefer large, nearby patches. This information will facilitate the development of a predictive framework to the study of bee movement and of models that predict the movement of genetically engineered pollen in bee-pollinated crops.

54 ENVIRONMENTAL SCIENCES↗

Downregulation of barley ferulate 5-hydroxylase dramatically alters straw lignin structure without impact on mechanical properties

Barley is a major cereal crop for temperate climates, and a diploid genetic model for polyploid wheat. Cereal straw biomass is an attractive source of feedstock for green technologies but lignin, a key determinant of feedstock recalcitrance, complicates bio-conversion processes. However, manipulating lignin content to improve the conversion process could negatively affect agronomic traits. An alternative approach is to manipulate lignin composition which influences the physical and chemical properties of straw. This study validates the function of a barley ferulate 5-hydroxylase gene and demonstrates that its downregulation using the RNA-interference approach substantially impacts lignin composition. We identified five barley genes having putative ferulate 5-hydroxylase activity. Downregulation of HvF5H1 substantially reduced the lignin syringyl/guaiacyl (S/G) ratio in straw while the lignin content, straw mechanical properties, plant growth habit, and grain characteristics all remained unaffected. Metabolic profiling revealed significant changes in the abundance of 173 features in the HvF5H1-RNAi lines. The drastic changes in the lignin polymer of transgenic lines highlight the plasticity of barley lignification processes and the associated potential for manipulating and improving lignocellulosic biomass as a feedstock for green technologies. On the other hand, our results highlight some differences between the lignin biosynthetic pathway in barley, a temperate climate grass, and the warm climate grass, rice, and underscore potential diversity in the lignin biosynthetic pathways in grasses.

59 BASIC BIOLOGICAL SCIENCES↗

Role of SaPCR2 in Zn Uptake in the Root Elongation Zone of the Zn/Cd Hyperaccumulator Sedum alfredii

Zn pollution is a potential toxicant for agriculture and the environment. Sedum alfredii is a Zn/Cd hyperaccumulator found in China and has been proven as a useful resource for the phytoremediation of Zn-contaminated sites. However, the molecular mechanism of Zn uptake in S. alfredii is limited. In this study, the function of SaPCR2 on Zn uptake in S. alfredii was identified by gene expression analysis, yeast function assays, Zn accumulation and root morphology analysis in transgenic lines to further elucidate the mechanisms of uptake and translocation of Zn in S. alfredii. The results showed that SaPCR2 was highly expressed in the root elongation zone of the hyperaccumulating ecotype (HE) S. alfredii, and high Zn exposure downregulated the expression of SaPCR2 in the HE S. alfredii root. The heterologous expression of SaPCR2 in yeast suggested that SaPCR2 was responsible for Zn influx. The overexpression of SaPCR2 in the non-hyperaccumulating ecotype (NHE) S. alfredii significantly increased the root uptake of Zn, but did not influence Mn, Cu or Fe. SR-μ-XRF technology showed that more Zn was distributed in the vascular buddle tissues, as well as in the cortex and epidermis in the transgenic lines. Root morphology was also altered after SaPCR2 overexpression, and a severe inhibition was observed. In the transgenic lines, the meristematic and elongation zones of the root were lower compared to the WT, and Zn accumulation in meristem cells was also reduced. These results indicate that SaPCR2 is responsible for Zn uptake, and mainly functions in the root elongation zone. This research on SaPCR2 could provide a theoretical basis for the use of genetic engineering technology in the modification of crops for their safe production and biological enhancement.

59 BASIC BIOLOGICAL SCIENCES↗