ANTHRACNOSE RESISTANCE GENE2 confers fungal resistance in sorghum.

Sorghum is an important food and feed crop globally; its production is hampered by anthracnose disease caused by the fungal pathogen Colletotrichum sublineola (Cs). Here, we report identification and characterization of ANTHRACNOSE RESISTANCE GENE 2 (ARG2) encoding a nucleotide-binding leucine-rich repeat (NLR) protein that confers race-specific resistance to Cs strains. ARG2 is one of a cluster of several NLR genes initially identified in the sorghum differential line SC328C that is resistant to some Cs strains. This cluster shows structural and copy number variations in different sorghum genotypes. Different sorghum lines carrying independent ARG2 alleles provided the genetic validation for the identity of the ARG2 gene. ARG2 expression is induced by Cs, and chitin induces ARG2 expression in resistant but not in susceptible lines. ARG2-mediated resistance is accompanied by higher expression of defense and secondary metabolite genes at early stages of infection, and anthocyanin and zeatin metabolisms are upregulated in resistant plants. Interestingly, ARG2 localizes to the plasma membrane when transiently expressed in Nicotiana benthamiana. Importantly, ARG2 plants produced higher shoot dry matter than near-isogenic lines carrying the susceptible allele suggesting an absence of an ARG2 associated growth trade-off. Furthermore, ARG2-mediated resistance is stable at a wide range of temperatures. Our observations open avenues for resistance breeding and for dissecting mechanisms of resistance.

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.

Ethylene-responsive SbWRKY50 suppresses leaf senescence by inhibition of chlorophyll degradation in sorghum.

The onset of leaf de-greening and senescence is governed by a complex regulatory network including environmental cues and internal factors such as transcription factors (TFs) and phytohormones, in which ethylene (ET) is one key inducer. However, the detailed mechanism of ET signalling for senescence regulation is still largely unknown. Here, we found that the WRKY TF SbWRKY50 from Sorghum bicolor L., a direct target of the key component ETHYLENE INSENSITIVE 3 in ET signalling, functioned for leaf senescence repression. The clustered regularly interspaced short palindromic repeats/CRISPR-associated protein9-edited SbWRKY50 mutant (SbWRKY5O-KO) of sorghum displayed precocious senescent phenotypes, while SbWRKY50 overexpression delayed age-dependent and dark-induced senescence in sorghum. SbWRKY50 negatively regulated chlorophyll degradation through direct binding to the promoters of several chlorophyll catabolic genes. In addition, SbWRKY50 recruited the Polycomb repressive complex 1 through direct interaction with SbBMI1A, to induce histone 2A mono-ubiquitination accumulation on the chlorophyll catabolic genes for epigenetic silencing and thus delayed leaf senescence. Especially, SbWRKY50 can suppress early steps of chlorophyll catabolic pathway via directly repressing SbNYC1 (NON-YELLOW COLORING 1). Other senescence-related hormones could also influence leaf senescence through repression of SbWRKY50. Hence, our work shows that SbWRKY50 is an essential regulator downstream of ET and SbWRKY50 also responds to other phytohormones for senescence regulation in sorghum.

Sorghum flour BRS 305 hybrid has the potential to modulate the intestinal microbiota of rats fed with a high-fat high-fructose diet.

AIM: The present study aimed to investigate the effect of dry heated whole sorghum BRS 305 hybrid flour on the gut microbiota modulation and gut health of rats fed with a high-fat high-fructose diet (HFHF). METHODS: In phase I (8 weeks), 45-50 days, male Wistar rats, were separated into the AIN93-M group (n = 10; fed with normal diet) and HFHF group (n = 20; fed with diet rich in saturated and simple carbohydrate). In phase II (10 weeks), we maintained the AIN-93-M group, and the HFHF group was divided into the HFHF group (n = 10) and HFHF plus sorghum flour group (n = 10). RESULTS: The consumption of sorghum flour increased the circular muscle layer and propionic acid when compared to the HFHF group. The sequencing of the 16S rRNA gene of the cecal microbiota presented no changes in the α-diversity and ß-diversity between. However, the sorghum group exhibited higher relative abundance of Firmicutes and higher Firmicutes/Bacteroidetes ratio compared to the other experimental groups, and lower abundance of Bacteroidetes, compared to the HFHF group. Despite, sorghum increased the abundance of the genera Roseburia and Lachnospiraceae_NK4A136_group compared to the HFHF group. No differences were observed in total goblet cell number, crypt thickness and height, circular muscle layer, secretory IgA, and butyric acid between all groups. CONCLUSIONS: The consumption of sorghum flour can modulate the gut microbiota composition, abundance of SCFA-producing bacteria, and intestinal morphology even with consumption of an HFHF diet.

Comprehensive analysis of MAPK cascade genes in sorghum (Sorghum bicolor L.) reveals SbMPK14 as a potential target for drought sensitivity regulation.

The mitogen-activated protein kinase (MAPK) cascade plays a crucial role in regulating many important biological processes in plants. Here, we identified and characterized eight MAPKK and 49 MAPKKK genes in sorghum and analyzed their differential expression under drought treatment; we also characterized 16 sorghum MAPK genes. RNA-seq analysis revealed that 10 MAPK cascade genes were involved in drought stress response at the transcriptome level in sorghum. Overexpression of SbMPK14 in Arabidopsis and maize resulted in hypersensitivity to drought by promoting water loss, indicating that SbMPK14 functions as a negative regulator of the drought response. Subsequent transcriptome analysis and qRT-PCR verification of maize SbMPK14 overexpression lines revealed that SbMPK14 likely increases plant drought sensitivity by suppressing the activity of specific ERF and WRKY transcription factors. This comprehensive study provides valuable insight into the mechanistic basis of MAPK cascade gene function and their responses to drought in sorghum.

Effect of Germinated Sorghum Extract on the Physical and Thermal Properties of Pre-Gelatinized Cereals, Sweet Potato and Beans Starches.

Starches from different botanical sources are affected in the presence of enzymes. This study investigated the impact of α-amylase on several properties of pre-gelatinized starches derived from chickpea (Cicer arietinum L.), wheat (Triticum aestivum L.), corn (Zea mays L.), white beans (Phaseolus vulgaris), and sweet potatoes (Ipomoea batatas L.). Specifically, the water holding capacity, freezable water content, sugar content, and water sorption isotherm (adsorption and desorption) properties were examined. The source of α-amylase utilized in this study was a germinated sorghum (Sorghum bicolor L. Moench) extract (GSE). The starch samples were subjected to annealing at temperatures of 40, 50, and 60 °C for durations of either 30 or 60 min prior to the process of gelatinization. A significant increase in the annealing temperature and GSE resulted in a notable enhancement in both the water-holding capacity and the sugar content of the starch. The ordering of starches in terms of their freezable water content is as follows: Chickpea starch (C.P.S) > white beans starch (W.B.S) > wheat starch (W.S) > chickpea starch (C.S) > sweet potato starch (S.P.S). The Guggenheim-Anderson-de Boer (GAB) model was only employed for fitting the data, as the Brunauer-Emmett-Teller (BET) model had a low root mean square error (RMSE). The application of annealing and GSE treatment resulted in a shift of the adsorption and desorption isotherms towards greater levels of moisture content. A strong hysteresis was found in the adsorption and desorption curves, notably within the water activity range of 0.6 to 0.8. The GSE treatment and longer annealing time had an impact on the monolayer water content (mo), as well as the C and K parameters of the GAB model, irrespective of the annealing temperature. These results can be used to evaluate the applicability of starch in the pharmaceutical and food sectors.

Genome-wide identification and analysis of ACP gene family in Sorghum bicolor (L.) Moench.

BACKGROUND: Acyl carrier proteins (ACP) constitute a very conserved carrier protein family. Previous studies have found that ACP not only takes part in the fatty acid synthesis process of almost all organisms, but also participates in the regulation of plant growth, development, and metabolism, and makes plants adaptable to stresses. However, this gene family has not been systematically studied in sorghum. RESULTS: Nine ACP family members were identified in the sorghum genome, which were located on chromosomes 1, 2, 5, 7, 8 and 9, respectively. Evolutionary analysis among different species divided the ACP family into four subfamilies, showing that the SbACPs were more closely related to maize. The prediction results of subcellular localization showed that SbACPs were mainly distributed in chloroplasts and mitochondria, while fluorescence localization showed that SbACPs were mainly localized in chloroplasts in tobacco leaf. The analysis of gene structure revealed a relatively simple genetic structure, that there were 1-3 introns in the sorghum ACP family, and the gene structure within the same subfamily had high similarity. The amplification method of SbACPs was mainly large fragment replication, and SbACPs were more closely related to ACPs in maize and rice. In addition, three-dimensional structure analysis showed that all ACP genes in sorghum contained four α helices, and the second helix structure was more conserved, implying a key role in function. Cis-acting element analysis indicated that the SbACPs might be involved in light response, plant growth and development regulation, biotic and abiotic stress response, plant hormone regulation, and other physiological processes. What's more, qRT-PCR analysis uncovered that some of SbACPs might be involved in the adaptive regulation of drought and salt stresses, indicating the close relationship between fatty acids and the resistance to abiotic stresses in sorghum. CONCLUSIONS: In summary, these results showed a comprehensive overview of the SbACPs and provided a theoretical basis for further studies on the biological functions of SbACPs in sorghum growth, development and abiotic stress responses.

Genome-Wide Identification of DOF Gene Family and the Mechanism Dissection of SbDof21 Regulating Starch Biosynthesis in Sorghum.

Starch is one of the main utilization products of sorghum (Sorghum bicolor L.), the fifth largest cereal crop in the world. Up to now, the regulation mechanism of starch biosynthesis is rarely documented in sorghum. In the present study, we identified 30 genes encoding the C2-C2 zinc finger domain (DOF), with one to three exons in the sorghum genome. The DOF proteins of sorghum were divided into two types according to the results of sequence alignment and evolutionary analysis. Based on gene expressions and co-expression analysis, we identified a regulatory factor, SbDof21, that was located on chromosome 5. SbDof21 contained two exons, encoding a 36.122 kD protein composed of 340 amino acids. SbDof21 co-expressed with 15 genes involved in the sorghum starch biosynthesis pathway, and the Pearson correlation coefficients (PCCs) with 11 genes were greater than 0.9. The results of qRT-PCR assays indicated that SbDof21 is highly expressed in sorghum grains, exhibiting low relative expression levels in the tissues of roots, stems and leaves. SbDOF21 presented as a typical DOF transcription factor (TF) that was localized to the nucleus and possessed transcriptional activation activity. Amino acids at positions 182-231 of SbDOF21 formed an important structure in its activation domain. The results of EMSA showed that SbDOF21 could bind to four tandem repeats of P-Box (TGTAAAG) motifs in vitro, such as its homologous proteins of ZmDOF36, OsPBF and TaPBF. Meanwhile, we also discovered that SbDOF21 could bind and transactivate SbGBSSI, a key gene in sorghum amylose biosynthesis. Collectively, the results of the present study suggest that SbDOF21 acts as an important regulator in sorghum starch biosynthesis, exhibiting potential values for the improvement of starch contents in sorghum.

Proteomic Analysis Revealed Different Molecular Mechanisms of Response to PEG Stress in Drought-Sensitive and Drought-Resistant Sorghums.

Drought is the major limiting factor that directly or indirectly inhibits the growth and reduces the productivity of sorghum (Sorghum bicolor (L.) Moench). As the main vegetative organ of sorghum, the response mechanism of the leaf to drought stress at the proteomic level has not been clarified. In the present study, nano-scale liquid chromatography mass spectrometry (nano-LC-MS/MS) technology was used to compare the changes in the protein expression profile of the leaves of drought-sensitive (S4 and S4-1) and drought-resistant (T33 and T14) sorghum varieties at the seedling stage under 25% PEG-6000 treatment for 24 h. A total of 3927 proteins were accurately quantitated and 46, 36, 35, and 102 differentially abundant proteins (DAPs) were obtained in the S4, S4-1, T14, and T33 varieties, respectively. Four proteins were randomly selected for parallel reaction monitoring (PRM) assays, and the results verified the reliability of the mass spectrometry (MS) results. The response mechanism of the drought-sensitive sorghum leaves to drought was attributed to the upregulation of proteins involved in the tyrosine metabolism pathway with defense functions. Drought-resistant sorghum leaves respond to drought by promoting the TCA cycle, enhancing sphingolipid biosynthesis, interfering with triterpenoid metabolite synthesis, and influencing aminoacyl-tRNA biosynthesis. The 17 screened important candidate proteins related to drought stress were verified by quantitative real-time PCR (qRT-PCR), the results of which were consistent with the results of the proteomic analysis. This study lays the foundation for revealing the drought-resistance mechanism of sorghum at the protein level. These findings will help us cultivate and improve new drought-resistant sorghum varieties.

Sequencing and Genomic Analysis of Sorghum DNA Introgression Variant Line R21 and Recipient Rice Jin Hui 1 Revealed Repetitive Element Variation.

Transferring the genome of distant species to crops is an efficient way to create new germplasms. However, the molecular mechanisms involved are unclear. In this study, a new rice restorer line R21 with heat tolerance was created by introgressing the genomic DNA of sorghum into the recipient restorer line Jin Hui 1. Assembly of rice R21 and Jin Hui 1 genomes was performed using PacBio sequencing technology. Comparative genome analysis and coverage statistics showed that the repetitive sequence atr0026 was a candidate introgression fragment of sorghum DNA. Sequence similarity analysis revealed that atr0026 was distributed at different copy numbers on the telomeric position of chromosomes 9 or 10 in R21, Jin Hui 1, and several rice varieties, indicating that the repetitive sequence from sorghum was highly conserved in rice. The repeat annotation in Gramineae indicated that ribosomal DNA loci that existed in atr0026 may be cause a rearrangement of chromosomes 9 and 10 of the R21 genome, resulting in a copy number variation at the 5' end of it. Our study lays the foundation for further elucidation of the molecular mechanisms underlying the heat tolerance of sorghum DNA introgression variant line R21, which is of great significance for guiding crop genetic breeding.

Molecular identification and expression analysis of five sucrose synthase genes in Sorghum Bicolor.

In higher plants, sucrose synthase (Susy, EC 2.4.1.13) as an enzyme with a core function, involved in the synthesis and breakdown of sugars, and plays an important role in growth and metabolism. Although, the different genes encoding Susy isozyme proteins have been cloned and functionally verified in several plant species, to date detailed information about the Susy genes is lacking in Sorghum. Here, we demonstrated the identification of five novel Susy genes from the sorghum genome database. Sequence, structure and phylogenetic analyses of these five SbSusy genes revealed evolutionary conservation through Susy gene family members across Sorghum and other crop plants. The expression of sorghum Susy genes was investigated via transcriptome database in various developmental stages and different tissues. Further qRT-PCR was performed to reveal the induction of SbSusy genes under salt, drought and sugar induction. The results indicated that all Susy genes were differentially expressed in various tissues and highly associated with sucrose metabolism. This study shows a theoretical reference of Susy genes in Sorghum, which provides new insights for the knowledge of the evolution relationships, and basic information to help clarify the molecular mechanism of Susy synthase genes in Sorghum. Supplementary Information: The online version contains supplementary material available at 10.1007/s12298-022-01166-8.

Genome-wide identification and characterization of the C2 domain family in Sorghum bicolor (L.) and expression profiles in response to saline-alkali stress.

The C2 domain family proteins in plants has been recently shown to be involved in the response to abiotic stress such as salt and drought stress. However, less information on C2 domain family members has been reported in Sorghum bicolor (L.), which is a tolerant cereal crop. To elaborate the mechanism of C2 domain family members in response to abiotic stress, bioinformatic methods were used to analyze this family. The results indicated that 69 C2 domain genes belonging to 5 different groups were first identified within the sorghum genome, and each group possessed various gene structures and conserved functional domains. Second, those C2 family genes were localized on 10 chromosomes 3 tandem repeat genes and 1 pair of repeat gene fragments were detected. The family members further presented a variety of stress responsive cis-elements. Third, in addition to being the major integral component of the membrane, sorghum C2 domain family proteins mainly played roles in response to abiotic and biotic stress with their organic transport and catalytic activity by specific location in the cell on the basis of gene ontology analysis. C2 family genes were differentially expressed in root, shoot or leaf, and shown different expression profiling after saline-alkali stress, which indicated that C2 family members played an important role in response to saline-alkali stress based on the transcription profiles of RNA-seq data and expression analysis by quantitative real-time polymerase chain reaction. Besides, most C2 family members were mainly located in cytoplasmi and nucleus. Weighted gene co-expression network analysis revealed three modules (turquoise, dark magenta and pink) that were associated with stress resistance, respectively. Therefore, the present research provides comprehensive information for further analysis of the molecular function of C2 domain family genes in sorghum. Supplementary Information: The online version contains supplementary material available at 10.1007/s12298-022-01222-3.

Genome-wide identification and expression analysis of phospholipase D gene in leaves of sorghum in response to abiotic stresses.

Abiotic stress caused by unsuitable environmental changes brings serious impacts on the growth and development of sorghum, resulting in significant loss in yield and quality every year. Phospholipase D is one of the key enzymes that catalyze the hydrolysis of phospholipids, and participates in plants response to abiotic stresses and phytohormones, whereas as the main producers of Phosphatidic acid (PA) signal, the detailed information about Phospholipase D associated (SbPLD) family in sorghum has been rarely reported. This study was performed to identify the PLD family gene in sorghum based on the latest genome annotation and to determine the expression of PLDs under abiotic stresses by qRT-PCR analysis. In this study, 13 PLD genes were identified in sorghum genome and further divided into 7 groups according to the phylogenetic analysis. All sorghum PLD family members harbored two conserved domains (HDK1&2) with catalytic activity, and most members contained a C2 domain. In ζ subfamily, C2 domain was replaced by PX and PH domain. The exon-intron structure of SbPLD genes within the same subfamily was highly conservative. The tissue specific expression analysis revealed different expression of SbPLD genes in various developmental stages. High level expression of SbPLDα3 was observed in almost all tissues, whereas SbPLDα4 was mainly expressed in roots. Under abiotic stress conditions, SbPLD genes responded actively to NaCl, ABA, drought (PEG) and cold (4 °C) treatment at the transcriptional level. The expression of SbPLDß1 was significantly up-regulated, while the transcription of SbPLDζ was suppressed under various stress conditions. In addition, SbPLDß1 and SbPLDδ2 were predicted to be the target genes of sbi-miR159 and sbi-miR167, respectively. This study will help to decipher the roles of PLDs in sorghum growth and abiotic stress responses. Supplementary Information: The online version contains supplementary material available at 10.1007/s12298-022-01200-9.

Genome-wide identification and expression profile analysis of trihelix transcription factor family genes in response to abiotic stress in sorghum [Sorghum bicolor (L.) Moench].

BACKGROUND: Transcription factors, including trihelix transcription factors, play vital roles in various growth and developmental processes and in abiotic stress responses in plants. The trihelix gene has been systematically studied in some dicots and monocots, including Arabidopsis, tomato, chrysanthemum, soybean, wheat, corn, rice, and buckwheat. However, there are no related studies on sorghum. RESULTS: In this study, a total of 40 sorghum trihelix (SbTH) genes were identified based on the sorghum genome, among which 34 were located in the nucleus, 5 in the chloroplast, 1 (SbTH38) in the cytoplasm, and 1 (SbTH23) in the extracellular membrane. Phylogenetic analysis of the SbTH genes and Arabidopsis and rice trihelix genes indicated that the genes were clustered into seven subfamilies: SIP1, GTγ, GT1, GT2, SH4, GTSb8, and orphan genes. The SbTH genes were located in nine chromosomes and none on chromosome 10. One pair of tandem duplication gene and seven pairs of segmental duplication genes were identified in the SbTH gene family. By qPCR, the expression of 14 SbTH members in different plant tissues and in plants exposed to six abiotic stresses at the seedling stage were quantified. Except for the leaves in which the genes were upregulated after only 2 h exposure to high temperature, the 12 SbTH genes were significantly upregulated in the stems of sorghum seedlings after 24 h under the other abiotic stress conditions. Among the selected genes, SbTH10/37/39 were significantly upregulated, whereas SbTH32 was significantly downregulated under different stress conditions. CONCLUSIONS: In this study, we identified 40 trihelix genes in sorghum and found that gene duplication was the main force driving trihelix gene evolution in sorghum. The findings of our study serve as a basis for further investigation of the functions of SbTH genes and providing candidate genes for stress-resistant sorghum breeding programmes and increasing sorghum yield.

Metabolic changes associated with differential salt tolerance in sorghum genotypes.

MAIN CONCLUSION: Accumulation of specific metabolites, mainly γ-aminobutyric acid, polyamines, and proline, was essential to homeostasis regulation and differential salt tolerance in sorghum genotypes. Salinity is severe abiotic stress that limits plant growth and development in arid and semi-arid regions. Survival to abiotic stresses depends on metabolic and sometimes even morphological adjustments. We measured the growth parameters, water relations, the content of ions (Na+, K+, Cl-), compatible solutes [some free amino acids (FAAs) including γ-aminobutyric acid (GABA) and proline and soluble carbohydrates) and polyamines (PAs), the activity of PAs metabolism enzymes, and metabolomic profile in plants after 14 days of salt stress treatment. These analyses were to evaluate the influence of metabolomic responses of sorghum genotypes exhibiting sensitivity (CSF18) or tolerance (CSF20) to salinity on plant growth. The salinity promoted growth reductions and induced increases in Na+ and Cl- content and decreases in K+ content. The water status and osmotic potential (Ψo) were reduced by salt stress, but to minimize damage, especially in the CSF20, the osmolytes and PAs contributed to the osmotic adjustment. The results showed that salinity induced an increase in putrescine (Put) in the sensitive genotype. However, it raised spermidine (Spd), spermine (Spm), and cadaverine (Cad) in the tolerant genotype. In addition, the regulation of polyamine oxidase can be related to Spm and GABA biosynthesis. Differential metabolic changes to salt tolerance include metabolites associated with tricarboxylic acid (TCA) cycle intermediates and the metabolisms of sugars, FAAs, and PAs.

Formation of root silica aggregates in sorghum is an active process of the endodermis.

Silica deposition in plants is a common phenomenon that correlates with plant tolerance to various stresses. Deposition occurs mostly in cell walls, but its mechanism is unclear. Here we show that metabolic processes control the formation of silica aggregates in roots of sorghum (Sorghum bicolor L.), a model plant for silicification. Silica formation was followed in intact roots and root segments of seedlings. Root segments were treated to enhance or suppress cell wall biosynthesis. The composition of endodermal cell walls was analysed by Raman microspectroscopy, scanning electron microscopy and energy-dispersive X-ray analysis. Our results were compared with in vitro reactions simulating lignin and silica polymerization. Silica aggregates formed only in live endodermal cells that were metabolically active. Silicic acid was deposited in vitro as silica onto freshly polymerized coniferyl alcohol, simulating G-lignin, but not onto coniferyl alcohol or ferulic acid monomers. Our results show that root silica aggregates form under tight regulation by endodermal cells, independently of the transpiration stream. We raise the hypothesis that the location and extent of silicification are primed by the chemistry and structure of polymerizing lignin as it cross-links to the wall.

Modelling tiller growth and mortality as a sink-driven process using Ecomeristem: implications for biomass sorghum ideotyping.

BACKGROUND AND AIMS: Plant modelling can efficiently support ideotype conception, particularly in multi-criteria selection contexts. This is the case for biomass sorghum, implying the need to consider traits related to biomass production and quality. This study evaluated three modelling approaches for their ability to predict tiller growth, mortality and their impact, together with other morphological and physiological traits, on biomass sorghum ideotype prediction. METHODS: Three Ecomeristem model versions were compared to evaluate whether tillering cessation and mortality were source (access to light) or sink (age-based hierarchical access to C supply) driven. They were tested using a field data set considering two biomass sorghum genotypes at two planting densities. An additional data set comparing eight genotypes was used to validate the best approach for its ability to predict the genotypic and environmental control of biomass production. A sensitivity analysis was performed to explore the impact of key genotypic parameters and define optimal parameter combinations depending on planting density and targeted production (sugar and fibre). KEY RESULTS: The sink-driven control of tillering cessation and mortality was the most accurate, and represented the phenotypic variability of studied sorghum genotypes in terms of biomass production and partitioning between structural and non-structural carbohydrates. Model sensitivity analysis revealed that light conversion efficiency and stem diameter are key traits to target for improving sorghum biomass within existing genetic diversity. Tillering contribution to biomass production appeared highly genotype and environment dependent, making it a challenging trait for designing ideotypes. CONCLUSIONS: By modelling tiller growth and mortality as sink-driven processes, Ecomeristem could predict and explore the genotypic and environmental variability of biomass sorghum production. Its application to larger sorghum genetic diversity considering water deficit regulations and its coupling to a genetic model will make it a powerful tool to assist ideotyping for current and future climatic scenario.

Systematic analysis of NAC transcription factors' gene family and identification of post-flowering drought stress responsive members in sorghum.

KEY MESSAGE: SbNAC genes (131) encoding 183 proteins were identified from the sorghum genome and characterized. The expression patterns of SbSNACs were evaluated at three sampling time points under post-flowering drought stress. NAC proteins are specific transcription factors in plants, playing vital roles in development and response to various environmental stresses. Despite the fact that Sorghum bicolor is well-known for its drought-tolerance, it suffers from grain yield loss due to pre and post-flowering drought stress. In the present study, 131 SbNAC genes encoding 183 proteins were identified from the sorghum genome. The phylogenetic trees were constructed based on the NAC domains of sorghum, and also based on sorghum with Arabidopsis and 8 known NAC domains of other plants, which classified the family into 15 and 19 subfamilies, respectively. Based on the obtained results, 13 SbNAC proteins joined the SNAC subfamily, and these proteins are expected to be involved in response to abiotic stresses. Promoter analysis revealed that all SbNAC genes comprise different stress-associated cis-elements in their promoters. UTRs analysis indicated that 101 SbNAC transcripts had upstream open reading frames, while 39 of the transcripts had internal ribosome entry sites in their 5'UTR. Moreover, 298 miRNA target sites were predicted to exist in the UTRs of SbNAC transcripts. The expression patterns of SbSNACs were evaluated in three genotypes at three sampling time points under post-flowering drought stress. Based on the results, it could be suggested that some gene members are involved in response to drought stress at the post-flowering stage since they act as positive or negative transcriptional regulators. Following further functional analyses, some of these genes might be perceived to be promising candidates for breeding programs to enhance drought tolerance in crops.

Short communication: Meta-analysis of dairy cows fed conventional sorghum or corn silages compared with brown midrib sorghum silage.

A meta-analysis was conducted to compare the effects of feeding dairy cows conventional sorghum silage (CSS) or conventional corn silage (CCS) compared with brown midrib sorghum silage (BMRSS) diets on dry matter intake (DMI), milk production, and milk composition. Data from 9 published articles (1984 to 2015) were used to contrast diets with CSS (7 means comparisons; 104 cows) or CCS (13 means comparisons; 204 cows) versus BMRSS diets. Statistical analysis was performed using fixed or random effects models with the Metafor package of R (https://www.R-project.org). The degree of heterogeneity was measured with the I2 statistic, and publication bias was determined with funnel plots and Egger's regression test. Other sources of heterogeneity of response were analyzed through meta-regression. Estimated effect size was calculated for DMI, milk production, and milk composition. No evidence of publication bias was observed for any variable tested. The highest degree of heterogeneity (I2 = 41.5 and 72.6%) was observed for DMI among dependent variables tested in both comparisons, indicating that intake responses to silage type are rather inconsistent; in contrast, milk production had the lowest degree of heterogeneity (I2 = 0%), supporting the idea that the responses of this variable to silage type were very consistent across studies. Compared with BMRSS diets, cows fed CSS diets exhibited decreased milk production (1.64 kg/d), milk fat concentration (0.09%), milk fat yield (0.08 kg/d), milk protein yield (0.04 kg/d), and milk lactose yield (0.16 kg/d) and tended to decrease DMI (0.83 kg/d). Compared with CCS diets, cows fed BMRSS diets increased milk fat concentration (0.10%), but decreased milk protein concentration (0.06%) and tended to increase lactose yield (0.08 kg/d). Meta-regression indicated that days in milk affected DMI and milk production when CSS diets were compared with BMRSS diets, and DMI when CCS diets were compared with BMRSS diets. Additionally, the inclusion rate of silage in the diet and dietary neutral detergent fiber affected yields of milk fat and lactose, respectively, when CCS and BMRSS diets were compared. Overall, lactation performance improved when cows were fed diets formulated with BMRSS compared with CSS, but performance was not different for cows fed BMRSS and CCS diets. However, the small sample size may have influenced these results by increasing the margin of the error and, concurrently, the power of the meta-analysis. Results of this analysis suggest that additional research is needed to explore the effects of days in milk and the inclusion rates of silages in the diets when comparing BMRSS with CSS or CCS.

Sorghum CCoAOMT and CCoAOMT-like gene evolution, structure, expression and the role of conserved amino acids in protein activity.

Sorghum is a crop plant that is grown for seeds, sucrose, forage and biofuel production. In all these applications, lignin is a superfluous component that decreases the efficiency of technological processes. Caffeoyl-coenzyme A O-methyltransferase (CCoAOMT) is an enzyme involved in monolignol synthesis that affects the efficiency of lignification and lignin composition. The sorghum genome harbors one CCoAOMT gene and six closely related CCoAOMT-like genes. The structures of four sorghum CCoAOMT-like enzymes suggest that these proteins might methylate caffeoyl coenzyme A and contribute to monolignol synthesis. In this study, two sorghum genes, CCoAOMT and one CCoAOMT-like, were found to be highly expressed in leaves, stems and immature seeds. The promoters of these genes possess clusters of transcription factor-binding sites specific for lignification, and this suggests that they are important for lignification. Phylogenetic analysis revealed that one sorghum CCoAOMT-like enzyme is closely related to ancestral cyanobacterial CCoAOMT-like proteins. The remaining CCoAOMT-like enzymes, including the one highly expressed in the leaves and stem, are closely related to CCoAOMT. Genes from these two groups possess different, evolutionarily conserved gene structures. The structure of the sorghum CCoAOMT-like protein from the ancestral clade was modeled and differences between enzymes from the two clades were analyzed. These results facilitate a better understanding of the evolution of genes involved in lignification, and provide valuable data for sorghum improvement through traditional breeding or molecular genetic techniques. The findings suggest that CCoAOMT-like genes might be recruited in lignification and raise questions of the frequency of such functional shifts.