Aphid-Induced Volatiles and Subsequent Attraction of Natural Enemies Varies among Sorghum Cultivars.

The production of herbivore-induced plant volatiles (HIPVs) is a type of indirect defense used by plants to attract natural enemies and reduce herbivory by insect pests. In many crops little is known about genotypic variation in HIPV production or how this may affect natural enemy attraction. In this study, we identified and quantified HIPVs produced by 10 sorghum (Sorghum bicolor) cultivars infested with a prominent aphid pest, the sorghum aphid (Melanaphis sorghi Theobald). Volatiles were collected using dynamic headspace sampling techniques and identified and quantified using GC-MS. The total amounts of volatiles induced by the aphids did not differ among the 10 cultivars, but overall blends of volatiles differed significantly in composition. Most notably, aphid herbivory induced higher levels of methyl salicylate (MeSA) emission in two cultivars, whereas in four cultivars, the volatile emissions did not change in response to aphid infestation. Dual-choice olfactometer assays were used to determine preference of the aphid parasitoid, Aphelinus nigritus, and predator, Chrysoperla rufilabris, between plants of the same cultivar that were un-infested or infested with aphids. Two aphid-infested cultivars were preferred by natural enemies, while four other cultivars were more attractive to natural enemies when they were free of aphids. The remaining four cultivars elicited no response from parasitoids. Our work suggests that genetic variation in HIPV emissions greatly affects parasitoid and predator attraction to aphid-infested sorghum and that screening crop cultivars for specific predator and parasitoid attractants has the potential to improve the efficacy of biological control.

[Characterization of sequences, expression profiling, and natural allelic variation analysis of the MYC gene family in sorghum (Sorghum bicolor)].

Sorghum aphid (Melanaphis sacchari) and head smut fungi (Sporisorium reilianum) infesting sorghum cause delayed growth and development, and reduce yield and quality. This study use bioinformatics and molecular biological approaches to profile the gene expression pattern during sorghum development and under pest infestation, and analyzed the natural allelic DNA variation of sorghum MYC gene family. The findings provide insights for potential application in breeding the stress resistant and high productivity sorghum varieties. The results indicated that there are 28 MYC genes identified in sorghum genome, distributed on 10 chromosomes. The bHLH_MYC_N and HLH domains are the conserved domains of the MYC gene in sorghum. Gene expression analysis showed that SbbHLH35.7g exhibited high expression levels in leaves, SbAbaIn showed strong expression in early grains, and SbMYC2.1g showed high expression levels in mature pollen. In anti-aphid strains at the 5-leaf stage, SbAbaIn, SbLHW.4g and SbLHW.2g were significantly induced in leaves, while SbbHLH35.7g displayed the highest expression level in panicle tissue, which was significantly induced by the infection of head smut. Promoter cis-element analysis identified methyl jasmonate (MJ), abscisic acid (ABA), salicylic acid (SA) and MYB-binding sites related to drought-stress inducibility. Furthermore, genomic resequencing data analysis revealed natural allelic DNA variations such as single nucleotide polymorphism (SNP) and insertion-deletion (INDEL) for the key SbMYCs. Protein interaction network analysis using STRING indicated that SbAbaIn interacts with TIFYdomain protein, and SbbHLH35.7g interacts with MDR and imporin. SbMYCs exhibited temporal and spatial expression patterns and played vital roles during the sorghum development. Infestation by sugarcane aphids and head smut fungi induced the expression of SbAbaIn and SbbHLH35.7g, respectively. SbAbaIn modulated the jasmonic acid (JA) pathway to regulate the expression of defensive genes, conferring resistance to insects. On the other hand, SbbHLH35.7g participated in detoxification reactions to defend against pathogens.

Nickel sources affect soil biological properties but do not affect sorghum growth.

Nickel (Ni) is an essential element, but it can be phytotoxic in high concentration, which may be caused by high availability in soil solution. The objective of this study was to evaluate the effect of sources and doses of Ni applied to a dystrophic Red Latosol cultivated with sorghum on i) the availability of the metal in the soil; ii) the impact on biological and biochemical properties of the soil; iii) the absorption and distribution in sorghum plants; and iv) crop productivity. The experiment was carried out within a completely randomized design with two nickel sources [nickel(II) nitrate, Ni(NO3)2 and nickel(III) oxide, Ni2O3], three doses (35, 70, and 140 mg Ni kg-1 soil), plus controls without Ni, with 3 replications. The concentrations of Ni in the soil, soil microbial biomass (SMB), basal soil respiration (BSR), metabolic quotient (qCO2), fluorescein diacetate (FDA) hydrolysis, and urease activity were determined. The concentrations of Ni in the leaf diagnostic and in the plant (shoot, root, and grains) were also measured. In the soil, the concentrations of available Ni remained between 0.21 and 54.01 mg Ni kg-1. Ni2O3 contributed very little to the increase in available Ni. SMB and the FDA hydrolysis were not affected by the Ni source or Ni dose, but BSR and qCO2 had significant increase with Ni application rates, suggesting the soil microorganisms faced stress. Soil urease activity was affected by Ni dose but not by Ni source. The dose of Ni as Ni(NO3)2 decreased the metal concentration in the plant, while that of Ni2O3 increased it. Nickel source did not affect dry mass production of the plants, but grain yield was affected in a dose-dependent manner when Ni2O3 was the source of Ni.

Non-destructive, whole-plant phenotyping reveals dynamic changes in water use efficiency, photosynthesis, and rhizosphere acidification of sorghum accessions under osmotic stress.

Noninvasive phenotyping can quantify dynamic plant growth processes at higher temporal resolution than destructive phenotyping and can reveal phenomena that would be missed by end-point analysis alone. Additionally, whole-plant phenotyping can identify growth conditions that are optimal for both above- and below-ground tissues. However, noninvasive, whole-plant phenotyping approaches available today are generally expensive, complex, and non-modular. We developed a low-cost and versatile approach to noninvasively measure whole-plant physiology over time by growing plants in isolated hydroponic chambers. We demonstrate the versatility of our approach by measuring whole-plant biomass accumulation, water use, and water use efficiency every two days on unstressed and osmotically stressed sorghum accessions. We identified relationships between root zone acidification and photosynthesis on whole-plant water use efficiency over time. Our system can be implemented using cheap, basic components, requires no specific technical expertise, and should be suitable for any non-aquatic vascular plant species.

Genotype by environment interaction, AMMI, GGE biplot, and mega environment analysis of elite Sorghum bicolor (L.) Moench genotypes in humid lowland areas of Ethiopia.

This study aimed to evaluate high-yielding, stable sorghum genotypes and determine the ideal (representative and discriminating) testing environments for genotypes in the humid lowlands of Ethiopia. A total of forty-two sorghum genotypes were used for a field trial conducted in six different environments using a randomized complete block design. Yield stability, Additive main effect, multiplicative interaction (AMMI), and genotype and genotype by environment interaction (GGE) were computed. The AMMI analysis explained 62.85% of the G×E variance. The AMMI1 biplot revealed that (G4; Mok079 and (G16; Ba066) genotypes had higher grain yields. AMMI2 biplot suggested that genotypes (G18; Y0470),(G23;100620), (G29; PML981475), and (G11; ETSC300373-4) show higher sensitivity to environmental changes because of their strong genotype-by-environment interactions. The GGE captured 79.46% of the GGE variance, and the GGE biplot identified genotypes (G4; Mok079), (G10; Sl081) and (G16; Ba066) were the most stable genotypes whereas(G39; ETSC120051-3) was the least stable genotypes. The GGE biplot identified Assosa (AS20) as a suitable environment, whereas PW20 and JM20 were the most discriminating and non-representative environments. The GGE biplot was found to identify three main mega-environments for sorghum growing in the humid lowlands of Ethiopia., both the AMMI and GGE biplots revealed (G4; Mok079) had the highest level of adaptability to all tested environments and was approved by the National Variety Release Committee for release in 2022.

Macronutrient application rescues performance of tolerant sorghum genotypes when infected by the parasitic plant striga.

BACKGROUND AND AIMS: Infection by the hemi-parasitic plant Striga hermonthica causes severe host-plant damage and seed production losses. Increased availability of essential plant nutrients reduces infection. Whether, how and to what extent, it also reduces striga-induced host-plant damage is not well studied. METHODS: Effects of improved macro- and micronutrient supply on host-plant performance under striga-free and infected conditions were investigated in greenhouse pot assays. One striga-sensitive and two striga-tolerant genotypes were compared. Plants growing in impoverished soils were supplied with (1) 25% of optimal macro- and micronutrients quantities, (2) 25% macro- and 100% micronutrients, (3) 100% macro- and 25% micronutrients, or (4) 100% of macro- and micronutrients. KEY RESULTS: Photosynthesis rates of striga-infected plants of the sensitive genotype increased with improved nutrition (12.2 to 22.1 µmol/m2/s1) but remained below striga-free levels (34.9-38.8 µmol/m2/s1). For the tolerant genotypes, increased macro-nutrient supply offset striga-induced photosynthesis losses. Striga-induced relative grain losses of 100% for the sensitive genotype were reduced to 74% by increased macronutrients. Grain losses of 80% in tolerant Ochuti, incurred at low nutrient supply, were reduced to 5% by improved nutrient supply. CONCLUSIONS: Increasing macro-nutrient supply reduces striga impact on host-plants but can only restore losses when applied to genotypes with a tolerant background.

Germination: A Powerful Way to Improve the Nutritional, Functional, and Molecular Properties of White- and Red-Colored Sorghum Grains.

This study explored the effects of the germination of red and white sorghum grains (Sorghum bicolor [Moench (L.)]) for up to seven days on various properties of the grain. Germination enriched sorghum's nutritional and sensory qualities while mitigating existing anti-nutritional factors. The study employed Fourier-transformed infrared spectroscopy (FT-IR) and scanning electron microscopy techniques to support its findings. Germination increased protein and lipid content but decreased starch content. White sorghum grains showed elevated calcium and magnesium but decreased iron, potassium, and zinc. Red sorghum grains showed a consistent decrease in mineral content during germination. Germination also increased fiber and lignin values in both sorghum varieties. The results of the FT-IR analysis demonstrate that germination induced significant changes in the molecular structure of white sorghum samples after 24 h, whereas this transformation was observed in red sorghum samples at four days. Total phenolic content (TPC) in red sorghum ranged from 136.64 ± 3.76 mg GAE/100 g to 379.5 ± 6.92 mg GAE/100 g. After 72 h of germination, the germinated seeds showed a threefold increase in TPC when compared to ungerminated seeds. Similarly, the TPC of white sorghum significantly increased (p < 0.05) from 52.84 ± 3.31 mg GAE/100 g to 151.76 mg GAE/100 g. Overall, during the 7-day germination period, all parameters showed an increase, and the germination process positively impacted the functional properties that contributed to the health benefits of white and red sorghum samples.

Overview of Cyanide Poisoning in Cattle from Sorghum halepense and S. bicolor Cultivars in Northwest Italy.

Sorghum plants naturally produce dhurrin, a cyanogenic glycoside that may be hydrolysed to cyanide, resulting in often-lethal toxicoses. Ruminants are particularly sensitive to cyanogenic glycosides due to the active role of rumen microbiota in dhurrin hydrolysis. This work provides an overview of a poisoning outbreak that occurred in 5 farms in Northwest Italy in August 2022; a total of 66 cows died, and many others developed acute toxicosis after being fed on either cultivated (Sorghum bicolor) or wild Sorghum (Sorghum halepense). Clinical signs were recorded, and all cows received antidotal/supportive therapy. Dead animals were subjected to necropsy, and dhurrin content was determined in Sorghum specimens using an LC-MS/MS method. Rapid onset, severe respiratory distress, recumbency and convulsions were the main clinical features; bright red blood, a bitter almond smell and lung emphysema were consistently observed on necropsy. The combined i.v. and oral administration of sodium thiosulphate resulted in a rapid improvement of clinical signs. Dhurrin concentrations corresponding to cyanide levels higher than the tolerated threshold of 200 mg/kg were detected in sorghum specimens from 4 out of 5 involved farms; thereafter, such levels declined, reaching tolerable concentrations in September-October. Feeding cattle with wild or cultivated Sorghum as green fodder is a common practice in Northern Italy, especially in summer. However, care should be taken in case of adverse climatic conditions, such as severe drought and tropical temperatures (characterising summer 2022), which are reported to increase dhurrin synthesis and storage.

Manifestation of agronomically valuable traits in the progeny of a sorghum mutant carrying the genetic construct for RNA silencing of the γ-kafirin gene.

Improving the nutritional value of grain sorghum, a drought- and heat-tolerant grain crop, is an important task in the context of global warming. One of the reasons for the low nutritional value of sorghum grain is the resistance of its storage proteins (kafirins) to proteolytic digestion, which is due, among other things, to the structural organization of protein bodies, in which γ-kafirin, the most resistant to proteases, is located on the periphery, encapsulating more easily digested α-kafirins. The introduction of genetic constructs capable of inducing RNA silencing of the γ-kafirin (gKAF1) gene opens up prospects for solving this problem. Using Agrobacterium-mediated genetic transformation of immature embryos of the grain sorghum cv. Avans we have obtained a mutant with improved digestibility of endosperm proteins (up to 92 %) carrying a genetic construct for RNA silencing of the gKAF1 gene. The goal of this work was to study the stability of inheritance of the introduced genetic construct in T2-T4 generations, to identify the number of its copies, as well as to trace the manifestation of agronomically valuable traits in the offspring of the mutant. The mutant lines were grown in experimental plots in three randomized blocks. The studied lines were characterized by improved digestibility of kafirins, a modified type of endosperm, completely or partially devoid of the vitreous layer, an increased percentage of lysine (by 75 %), reduced plant height, peduncle length, 1000-grains weight, and grain yield from the panicle. In T2, a line with monogenic control of GA resistance was selected. qPCR analysis showed that in different T3 and T4 plants, the genetic construct was present in 2-4 copies. In T3, a line with a high digestibility of endosperm proteins (81 %) and a minimal decrease in agronomically valuable traits (by 5-7 %) was selected.

Data-Driven Characterization of Metabolome Reprogramming during Early Development of Sorghum Seedlings.

Specialized metabolites are produced via discrete metabolic pathways. These small molecules play significant roles in plant growth and development, as well as defense against environmental stresses. These include damping off or seedling blight at a post-emergence stage. Targeted metabolomics was followed to gain insights into metabolome changes characteristic of different developmental stages of sorghum seedlings. Metabolites were extracted from leaves at seven time points post-germination and analyzed using ultra-high performance liquid chromatography coupled to mass spectrometry. Multivariate statistical analysis combined with chemometric tools, such as principal component analysis, hierarchical clustering analysis, and orthogonal partial least squares-discriminant analysis, were applied for data exploration and to reduce data dimensionality as well as for the selection of potential discriminant biomarkers. Changes in metabolome patterns of the seedlings were analyzed in the early, middle, and late stages of growth (7, 14, and 29 days post-germination). The metabolite classes were amino acids, organic acids, lipids, cyanogenic glycosides, hormones, hydroxycinnamic acid derivatives, and flavonoids, with the latter representing the largest class of metabolites. In general, the metabolite content showed an increase with the progression of the plant growth stages. Most of the differential metabolites were derived from tryptophan and phenylalanine, which contribute to innate immune defenses as well as growth. Quantitative analysis identified a correlation of apigenin flavone derivatives with growth stage. Data-driven investigations of these metabolomes provided new insights into the developmental dynamics that occur in seedlings to limit post-germination mortality.

Expression, purification and crystallization of the photosensory module of phytochrome B (phyB) from Sorghum bicolor.

Sorghum, a short-day tropical plant, has been adapted for temperate grain production, in particular through the selection of variants at the MATURITY loci (Ma1-Ma6) that reduce photoperiod sensitivity. Ma3 encodes phytochrome B (phyB), a red/far-red photochromic biliprotein photoreceptor. The multi-domain gene product, comprising 1178 amino acids, autocatalytically binds the phytochromobilin chromophore to form the photoactive holophytochrome (Sb.phyB). This study describes the development of an efficient heterologous overproduction system which allows the production of large quantities of various holoprotein constructs, along with purification and crystallization procedures. Crystals of the Pr (red-light-absorbing) forms of NPGP, PGP and PG (residues 1-655, 114-655 and 114-458, respectively), each C-terminally tagged with His6, were successfully produced. While NPGP crystals did not diffract, those of PGP and PG diffracted to 6 and 2.1 Šresolution, respectively. Moving the tag to the N-terminus and replacing phytochromobilin with phycocyanobilin as the ligand produced PG crystals that diffracted to 1.8 Šresolution. These results demonstrate that the diffraction quality of challenging protein crystals can be improved by removing flexible regions, shifting fusion tags and altering small-molecule ligands.

Cover Crop Root Exudates Impact Soil Microbiome Functional Trajectories in Agricultural Soils.

Background: Cover cropping is an agricultural practice that uses secondary crops to support the growth of primary crops through various mechanisms including erosion control, weed suppression, nutrient management, and enhanced biodiversity. Cover crops may elicit some of these ecosystem services through chemical interactions with the soil microbiome via root exudation, or the release of plant metabolites from roots. Phytohormones are one metabolite type exuded by plants that activate the rhizosphere microbiome, yet managing this chemical interaction remains an untapped mechanism for optimizing plant-soil microbiome interactions. Currently, there is limited understanding on the diversity of cover crop phytohormone root exudation patterns and how these chemical messages selectively enrich specific microbial taxa and functionalities in agricultural soils. Results: Here, we link variability in cover crop root exudate composition to changes in soil microbiome functionality. Exudate chemical profiles from 4 cover crop species (Sorghum bicolor, Vicia villosa, Brassica napus, and Secale cereal) were used as the chemical inputs to decipher microbial responses. These distinct exudate profiles, along with a no exudate control, were amended to agricultural soil microcosms with microbial responses tracked over time using metabolomes and genome-resolved metatranscriptomes. Our findings illustrated microbial metabolic patterns were unique in response to cover crop exudate inputs over time, particularly by sorghum and cereal rye amended microcosms where we identify novel microbial members (at the genera and family level) who produced IAA and GA4 over time. We also identify broad changes in microbial nitrogen cycling in response chemical inputs. Conclusions: We highlight that root exudate amendments alter microbial community function and phytohormone metabolisms, particularly in response to root exudates isolated from cereal rye and sorghum plants. Additionally, we constructed a soil microbial genomic catalog of microorganisms responding to commonly used cover crops, a public resource for agriculturally-relevant microbes. Many of our exudate-stimulated microorganisms are representatives from poorly characterized or novel taxa, highlighting the yet to be discovered metabolic reservoir harbored in agricultural soils. Our findings emphasize the tractability of high-resolution multiomics approaches to investigate processes relevant for agricultural soils, opening the possibility of targeting specific soil biogeochemical outcomes through biological precision agricultural practices that use cover crops and the microbiome as levers for enhanced crop production.

Differential physiological and production responses of C3 and C4 crops to climate factor interactions.

This study examined the effect of the interactions of key factors associated with predicted climate change (increased temperature, and drought) and elevated CO2 concentration on C3 and C4 crop representatives, barley and sorghum. The effect of two levels of atmospheric CO2 concentration (400 and 800 ppm), three levels of temperature regime (21/7, 26/12 and 33/19°C) and two regimes of water availability (simulation of drought by gradual reduction of irrigation and well-watered control) in all combinations was investigated in a pot experiment within growth chambers for barley variety Bojos and sorghum variety Ruby. Due to differences in photosynthetic metabolism in C3 barley and C4 sorghum, leading to different responses to elevated CO2 concentration, we hypothesized mitigation of the negative drought impact in barley under elevated CO2 concentration and, conversely, improved performance of sorghum at high temperatures. The results demonstrate the decoupling of photosynthetic CO2 assimilation and production parameters in sorghum. High temperatures and elevated CO2 concentration resulted in a significant increase in sorghum above- and below-ground biomass under sufficient water availability despite the enhanced sensitivity of photosynthesis to high temperatures. However, the negative effect of drought is amplified by the effect of high temperature, similarly for biomass and photosynthetic rates. Sorghum also showed a mitigating effect of elevated CO2 concentration on the negative drought impact, particularly in reducing the decrease of relative water content in leaves. In barley, no significant factor interactions were observed, indicating the absence of mitigating the negative drought effects by elevated CO2 concentration. These complex interactions imply that, unlike barley, sorghum can be predicted to have a much higher variability in response to climate change. However, under conditions combining elevated CO2 concentration, high temperature, and sufficient water availability, the outperforming of C4 crops can be expected. On the contrary, the C3 crops can be expected to perform even better under drought conditions when accompanied by lower temperatures.

Screening of cadmium resistant bacteria and their growth promotion of Sorghum bicolor (L.) Moench under cadmium stress.

Heavy metal pollution of agricultural soils, especially from cadmium (Cd) contaminationcaused serious problems in both food security and economy. Sorghum bicolor (L.) showed a great potential in phytoremediation of Cd contamination due to its fast growth, high yield and easy harvesting. However, the growth of S. bicolor plants tends to be inhibited under Cd exposure, which limited its application for Cd remediation. Plant growth-promoting rhizobacteria may enhance the Cd resistance of S. bicolor and thus improve its Cd removal efficiency. In this study, three Cd-resistant bacteria were screened based on Cd and acid tolerance and identified as Bacillus velezensis QZG6, Enterobacter cloacae QZS3 and Bacillus cereus QZS8, by 16S rRNA sequencing. Inoculation of hydroponic plants with strains QZG6, QZS3 or QZS8 significantly promoted the biomass of sorghum plants by 31.52%, 50.20% and 26.93%, respectively, compared with those of uninoculated plants under Cd exposure. The activity of SOD, POD and MDA content in Cd-stressed S. bicolor plants were reduced of 65.74%, 31.52%, and 80.91%, respectively, when inoculated with the strains QZS3. For pot experiment, strains QZG6, QZS3 and QZS8 significantly promoted the biomass of sorghum plants by 47.30%, 19.27% and 58.47%, compared with those of uninoculated plants under Cd exposure. The activity of SOD, POD and MDA content in Cd-stressed S. bicolor plants were reduced of 67.20%, 22.40%, and 40.65%, respectively, when inoculated with the strains QZS3. All these three strains significantly increased the Cd removal efficiency of the plants by 42.16% (QZG6), 18.76% (QZS3) and 21.06% (QZS8). To investigate the bacterial characteristics associated with growth promotion of S. bicolor plants, the ability on nitrogen fixation, phosphorus solubilization, siderophores production, and phytohormones production were determined. All the strains were able to fix nitrogen. Phosphorus release was observed for strains QZG6 (inorganic or organic phosphorus) and QZS3 (inorganic phosphorus). Both QZG6 and QZS8 were able to produce siderophores, while only QZG6 was positive for ACC deaminase. All the strains produced IAA, SA and GA. These results indicated that the three strains promoted the plant growth under Cd stress, probably through Cd detoxification by siderophores, as well as through growth regulation by N/P nutrient supply and phytohormone. The present study showed a great potential of the three Cd-resistant strains combined with S. bicolor plants in the remediation of Cd-polluted soils, which may provide a new insight into combining the advantages of microbes and plants to improve the remediation of Cd-contaminated soils.

Identification of transcriptional networks controlling leaf sheath growth in Sorghum bicolor.

OBJECTIVES: The objective of this data set was to identify transcriptional networks that control elongation of seedling leaf sheaths in the C4 grass Sorghum bicolor. One motivation was that leaf sheaths are a primary constituent of stems in grass seedlings; therefore, genes that control growth of this organ are important contributors to successful transition from the seedling stage to the mature plant stage and, ultimately, crop success. Since diurnal rhythms contribute to regulation of signaling networks responsible for growth, a time course representing the late afternoon and early evening was anticipated to pinpoint important control genes for stem growth. Ultimately, the expected outcome was discovery of transcript networks that integrate internal and external signals to fine tune leaf sheath growth and, consequently, plant height. DATA DESCRIPTION: The data set is RNAseq profiling of upper leaf sheaths collected from wild type Sorghum bicolor (BTx623 line) plants at four-hour intervals from 12.5 h after dawn to 20 h after dawn. Global transcript levels in leaves were determined by deep sequencing of mRNA from four individual seedlings at each time point. This data set contains sequences representing the spectrum of mRNAs from individual genes. This data set enables detection of significant changes in gene-level expression caused by the progression of the day from late afternoon to the middle of the night. This data set is useful to identify gene expression networks regulating growth in the leaf sheath, an organ that is a major contributor to the sorghum seedling stem and defines seedling height.

A normalization method that controls for total RNA abundance affects the identification of differentially expressed genes, revealing bias toward morning-expressed responses.

RNA-Sequencing is widely used to investigate changes in gene expression at the transcription level in plants. Most plant RNA-Seq analysis pipelines base the normalization approaches on the assumption that total transcript levels do not vary between samples. However, this assumption has not been demonstrated. In fact, many common experimental treatments and genetic alterations affect transcription efficiency or RNA stability, resulting in unequal transcript abundance. The addition of synthetic RNA controls is a simple correction that controls for variation in total mRNA levels. However, adding spike-ins appropriately is challenging with complex plant tissue, and carefully considering how they are added is essential to their successful use. We demonstrate that adding external RNA spike-ins as a normalization control produces differences in RNA-Seq analysis compared to traditional normalization methods, even between two times of day in untreated plants. We illustrate the use of RNA spike-ins with 3' RNA-Seq and present a normalization pipeline that accounts for differences in total transcriptional levels. We evaluate the effect of normalization methods on identifying differentially expressed genes in the context of identifying the effect of the time of day on gene expression and response to chilling stress in sorghum.

Haplotypes at the sorghum ARG4 and ARG5 NLR loci confer resistance to anthracnose.

Sorghum anthracnose caused by the fungus Colletotrichum sublineola (Cs) is a damaging disease of the crop. Here, we describe the identification of ANTHRACNOSE RESISTANCE GENES (ARG4 and ARG5) encoding canonical nucleotide-binding leucine-rich repeat (NLR) receptors. ARG4 and ARG5 are dominant resistance genes identified in the sorghum lines SAP135 and P9830, respectively, that show broad-spectrum resistance to Cs. Independent genetic studies using populations generated by crossing SAP135 and P9830 with TAM428, fine mapping using molecular markers, comparative genomics and gene expression studies determined that ARG4 and ARG5 are resistance genes against Cs strains. Interestingly, ARG4 and ARG5 are both located within clusters of duplicate NLR genes at linked loci separated by ~1 Mb genomic region. SAP135 and P9830 each carry only one of the ARG genes while having the recessive allele at the second locus. Only two copies of the ARG5 candidate genes were present in the resistant P9830 line while five non-functional copies were identified in the susceptible line. The resistant parents and their recombinant inbred lines carrying either ARG4 or ARG5 are resistant to strains Csgl1 and Csgrg suggesting that these genes have overlapping specificities. The role of ARG4 and ARG5 in resistance was validated through sorghum lines carrying independent recessive alleles that show increased susceptibility. ARG4 and ARG5 are located within complex loci displaying interesting haplotype structures and copy number variation that may have resulted from duplication. Overall, the identification of anthracnose resistance genes with unique haplotype stucture provides a foundation for genetic studies and resistance breeding.

Sorghum hybrids grown in hydroponics contrast for phosphorus use efficiency / Híbridos de sorgo crescidos em hidroponia contrastam para eficiência de uso de fósforo

Phosphorus (P) use efficiency is crucial for sorghum production. P acquisition efficiency is the most important component of P use efficiency. The early-stage evaluation of plant development is a useful tool for identifying P-efficient genotypes. This study aimed to identify sorghum hybrids that are efficient in P use efficiency and assess the genetic diversity among hybrids based on traits related to P acquisition efficiency. Thus, 38 sorghum hybrids and two inbred lines (checks) were evaluated under low and high P in a paper pouch system with nutrient solution. Biomass and root traits related to P efficiency were measured. There was no interaction between genotypes and P levels concerning all evaluated traits. The biomass and root traits, except root diameter, presented smaller means under low P than high P. Efficient and inefficient hybrids under each P level were identified. The genetic diversity assessment grouped these genotypes in different clusters. The hybrids AG1090, MSK326, AG1060, 1G100, AS 4639, DKB 540, and DKB 590 were superior under low-P and high-P. Hybrids SC121, 1236020 e 1167017 presented the lowest means than all other hybrids, under both conditions. The evaluated hybrids showed phenotypic diversity for traits related to P acquisition, such as root length and root surface area, which can be useful for establishing selection strategies for sorghum breeding programs and increasing P use efficiency.

A eficiência do uso do fósforo (P) é fundamental para a produção de sorgo. A avaliação no estágio inicial do desenvolvimento da planta é uma ferramenta útil para a identificação de genótipos eficientes de P. Este trabalho teve como objetivo identificar híbridos de sorgo que sejam eficientes ao uso de P e avaliar a diversidade genética entre os híbridos com base em características relacionadas à eficiência de aquisição de P. Assim, 38 híbridos de sorgo e duas linhagens (testemunhas) foram avaliados sob baixo e alto P em sistema de pastas de papel com solução nutritiva. Características de biomassa e de raiz relacionadas à eficiência de P foram mensuradas. Não houve interação entre genótipos e níveis de P em todas as características avaliadas. As características de biomassa e raiz, exceto o diâmetro da raiz, apresentaram médias menores sob baixo P em comparação com alto P. Híbridos eficientes e ineficientes sob cada nível de P foram identificados e agrupados quanto à diversidade genética. Os híbridos AG1090, MSK326, AG1060, 1G100, AS 4639, DKB 540 e DKB 590 foram superiores sob baixo-P e alto-P. Os híbridos SC121, 1236020 e 1167017 apresentaram as menores médias que todos os outros híbridos, em ambas condições. Os híbridos avaliados apresentaram diversidade fenotípica para características relacionadas à aquisição de P, como comprimento e área superficial da raiz, o que pode ser útil para estabelecer estratégias de seleção para programas de melhoramento de sorgo e aumentar a eficiência de uso do P.

Sorghum hybrids grown in hydroponics contrast for phosphorus use efficiency

Abstract Phosphorus (P) use efficiency is crucial for sorghum production. P acquisition efficiency is the most important component of P use efficiency. The early-stage evaluation of plant development is a useful tool for identifying P-efficient genotypes. This study aimed to identify sorghum hybrids that are efficient in P use efficiency and assess the genetic diversity among hybrids based on traits related to P acquisition efficiency. Thus, 38 sorghum hybrids and two inbred lines (checks) were evaluated under low and high P in a paper pouch system with nutrient solution. Biomass and root traits related to P efficiency were measured. There was no interaction between genotypes and P levels concerning all evaluated traits. The biomass and root traits, except root diameter, presented smaller means under low P than high P. Efficient and inefficient hybrids under each P level were identified. The genetic diversity assessment grouped these genotypes in different clusters. The hybrids AG1090, MSK326, AG1060, 1G100, AS 4639, DKB 540, and DKB 590 were superior under low-P and high-P. Hybrids SC121, 1236020 e 1167017 presented the lowest means than all other hybrids, under both conditions. The evaluated hybrids showed phenotypic diversity for traits related to P acquisition, such as root length and root surface area, which can be useful for establishing selection strategies for sorghum breeding programs and increasing P use efficiency.

Resumo A eficiência do uso do fósforo (P) é fundamental para a produção de sorgo. A avaliação no estágio inicial do desenvolvimento da planta é uma ferramenta útil para a identificação de genótipos eficientes de P. Este trabalho teve como objetivo identificar híbridos de sorgo que sejam eficientes ao uso de P e avaliar a diversidade genética entre os híbridos com base em características relacionadas à eficiência de aquisição de P. Assim, 38 híbridos de sorgo e duas linhagens (testemunhas) foram avaliados sob baixo e alto P em sistema de pastas de papel com solução nutritiva. Características de biomassa e de raiz relacionadas à eficiência de P foram mensuradas. Não houve interação entre genótipos e níveis de P em todas as características avaliadas. As características de biomassa e raiz, exceto o diâmetro da raiz, apresentaram médias menores sob baixo P em comparação com alto P. Híbridos eficientes e ineficientes sob cada nível de P foram identificados e agrupados quanto à diversidade genética. Os híbridos AG1090, MSK326, AG1060, 1G100, AS 4639, DKB 540 e DKB 590 foram superiores sob baixo-P e alto-P. Os híbridos SC121, 1236020 e 1167017 apresentaram as menores médias que todos os outros híbridos, em ambas condições. Os híbridos avaliados apresentaram diversidade fenotípica para características relacionadas à aquisição de P, como comprimento e área superficial da raiz, o que pode ser útil para estabelecer estratégias de seleção para programas de melhoramento de sorgo e aumentar a eficiência de uso do P.

Structural and functional characteristics and expression profile of the 20S proteasome gene family in Sorghum under abiotic stress.

The 26S proteasome is a molecular machine that catalyzes and degrades protein intracellularly with the help of its core complex called 20S proteasome. The 20S proteasomes degrade and cleave denatured, cytotoxic, damaged, and unwanted proteins via proteolysis and impart biotic and abiotic stress tolerance in model plants. This study identified 20 genes, namely, 10 SbPA and 10 SbPB that encode for α- and ß-subunits of the 20S proteasome in Sorghum bicolor (L.) Moench (2n= 20). These genes have been found distributed on the 1st, 2nd, 3rd, 4th, 5th, 7th, and 10th chromosomes. These sorghum genes were orthologous to corresponding rice. Phylogenetic analysis clustered these genes into seven clades, each with one of the seven α-subunits (1 to 7) and one of the seven ß-subunits (1 to 7). In silico gene expression analysis suggested that nine genes were involved in abiotic stress response (cold, drought, and abscisic acid hormone). The expression of these proteasomal genes was studied in shoots and roots exposed to different abiotic stresses (cold, drought, and abscisic acid) by quantitative real-time polymerase chain reaction. A significant increase in the relative fold expression of SbPBA1, SbPAA1, SbPBG1, SbPBE1, and SbPAG1 genes under ABA and drought stress provides an insight into its involvement in abiotic stress. No expression was observed for cold stress of these genes indicating their non-involvement. It is believed that additional investigation into the SbPA/SbPB genes would aid in the creation of S. bicolor cultivars that are resistant to climate change.