Comprehensive Genomics and Proteomics Analysis Reveals the Multiple Response Strategies of Endophytic Bacillus sp. WR13 to Iron Limitation.

Iron (Fe) is an important metal element for the growth of bacteria. Many bacteria respond to Fe limitation through a variety of strategies. We previously isolated an endophyte Bacillus sp. WR13 from wheat root. However, whether and how this strain can cope with Fe-deficient environments remains unclear. In this study, the growth of WR13 under Fe starvation was investigated, and the underlying mechanisms of WR13 in response to Fe starvation were elucidated via genomics and iTRAQ-based proteomics. Under Fe limitation, WR13 showed a growth pattern similar to that of Fe sufficiency. Genomics analysis demonstrated that WR13 had gene clusters related to siderophore synthesis (dhbACEBF), transportation (bcbE), uptake (feuABC-yusV) and hydrolysis (besA). These genes were significantly up-regulated in Fe-starved WR13, which resulted in more siderophore production. Proteomics data revealed that many Fe-containing proteins such as ACO, HemQ, ferredoxin, CNP, and SufD were significantly reduced under Fe limitation. Meanwhile, significant decreases in many proteins involved in glycolysis, TCA cycle, pentose phosphate pathway; asparagine, glutamine, methionine, and serine metabolism; and phospholipid hydrolysis were also observed. Overall, this study shows that Bacillus sp. WR13 was able to respond to Fe limitation via multiple strategies and provides a theoretical basis for the application of WR13 in Fe-deficient soil.

Selection of Reference Genes for Normalization of qRT‒PCR Analysis in the Soybean Aphid Aphis glycines Matsumura (Hemiptera: Aphididae).

The soybean aphid Aphis glycines Matsumura is a predominant insect pest in Asia and North America and causes great losses to soybean. The release of genome data for A. glycines will facilitate gene function research in the future. However, suitable reference genes for A. glycines under various experimental conditions are scarce. To search for appropriate reference genes for A. glycines, nine candidate reference genes, including Act, α-Tub, ß-Tub, RPS12, RPS18, RPL5, RPL27, EF1α, and Fer, were tested under six experimental conditions to evaluate their suitability for use in the normalization of qRT‒PCR data. Results showed that EF1α and RPS12 were optimal for the developmental stages of A. glycines, RPS18 and RPS12 were appropriate for wing dimorphism, ß-Tub and RPS18 were suitable for different tissues and RPL5, and α-Tub could be used for normalization at different temperatures. ß-Tub and EF1α could be proposed as reference genes for insecticide treatment, and RPL5 and RPS12 were found to be the most stable reference genes in different photoperiods. The results provide appropriate reference genes for analyzing gene expression in A. glycines and contribute to future research on the molecular physiology and biochemistry of A. glycines.

Halotolerant Bacillus altitudinis WR10 improves salt tolerance in wheat via a multi-level mechanism.

Soil salinity is an important abiotic stress factor that seriously affects the crop growth and yield. Use of plant-derived microorganisms is a promising strategy to alleviate salt stress. In a previous study, the endophytic strain Bacillus altitudinis WR10 isolated from wheat roots showed high salt resistance. In this study, we investigated the efficacy of WR10 in improving the salt tolerance of wheat and its potential mechanisms using a hydroponic test. Under salt stress, WR10 inoculation significantly increased the lengths and dry weights of the roots and shoots, indicating that WR10 improves wheat salt tolerance at the seedling stage. WR10 inoculation significantly reduced Na+ accumulation and enhanced K+, P, and Ca2+ uptake in salt-stressed plants, which can be attributed to the upregulated gene expression of H+-ATPase as well as the P-solubilizing and biofilm-producing characteristics of WR10. At the transcriptional level, L-ascorbate peroxidase (APX), glutathione (GSH) synthetase related to GSH biosynthesis, and phenylpropanoid biosynthesis genes (CYP73A, 4CL, and CAD) were significantly upregulated, whereas those of GSH metabolism genes (glutathione S-transferase and gamma-glutamyltranspeptidase) were significantly downregulated in WR10-applied wheat roots under salt stress. These changes increased the APX activity and GSH levels and resulted in a decrease in hydrogen peroxide levels. Additionally, a decrease in proline content was observed in WR10-inoculated plants under salt stress because of WR10-induced upregulation of proline dehydrogenase gene expression. These results provide supporting evidence that WR10 improves wheat salt tolerance via more than one mechanism and open a window of opportunity for WR10 application in salinized soil.

Arms Race between the Host and Pathogen Associated with Fusarium Head Blight of Wheat.

Fusarium head blight (FHB), or scab, caused by Fusarium species, is an extremely destructive fungal disease in wheat worldwide. In recent decades, researchers have made unremitting efforts in genetic breeding and control technology related to FHB and have made great progress, especially in the exploration of germplasm resources resistant to FHB; identification and pathogenesis of pathogenic strains; discovery and identification of disease-resistant genes; biochemical control, and so on. However, FHB burst have not been effectively controlled and thereby pose increasingly severe threats to wheat productivity. This review focuses on recent advances in pathogenesis, resistance quantitative trait loci (QTLs)/genes, resistance mechanism, and signaling pathways. We identify two primary pathogenetic patterns of Fusarium species and three significant signaling pathways mediated by UGT, WRKY, and SnRK1, respectively; many publicly approved superstar QTLs and genes are fully summarized to illustrate the pathogenetic patterns of Fusarium species, signaling behavior of the major genes, and their sophisticated and dexterous crosstalk. Besides the research status of FHB resistance, breeding bottlenecks in resistant germplasm resources are also analyzed deeply. Finally, this review proposes that the maintenance of intracellular ROS (reactive oxygen species) homeostasis, regulated by several TaCERK-mediated theoretical patterns, may play an important role in plant response to FHB and puts forward some suggestions on resistant QTL/gene mining and molecular breeding in order to provide a valuable reference to contain FHB outbreaks in agricultural production and promote the sustainable development of green agriculture.

Identification of CDPKs involved in TaNOX7 mediated ROS production in wheat.

As the critical sensors and decoders of calcium signal, calcium-dependent protein kinase (CDPK) has become the focus of current research, especially in plants. However, few resources are available on the properties and functions of CDPK gene family in Triticum aestivum (TaCDPK). Here, a total of 79 CDPK genes were identified in the wheat genome. These TaCDPKs could be classified into four subgroups on phylogenesis, while they may be classified into two subgroups based on their tissue and organ-spatiotemporal expression profiles or three subgroups according to their induced expression patterns. The analysis on the signal network relationships and interactions of TaCDPKs and NADPH (reduced nicotinamide adenine dinucleotide phosphate oxidases, NOXs), the key producers for reactive oxygen species (ROS), showed that there are complicated cross-talks between these two family proteins. Further experiments demonstrate that, two members of TaCDPKs, TaCDPK2/4, can interact with TaNOX7, an important member of wheat NOXs, and enhanced the TaNOX7-mediated ROS production. All the results suggest that TaCDPKs are highly expressed in wheat with distinct tissue or organ-specificity and stress-inducible diversity, and play vital roles in plant development and response to biotic and abiotic stresses by directly interacting with TaNOXs for ROS production.

Comparative Transcriptome Analysis of Male Sterile Anthers Induced by High Temperature in Wheat (Triticum aestivum L.).

Global warming will have a negative effect on agricultural production as high temperature (HT) stress can seriously threaten plant growth and reproduction. Male sterility caused by HT may be exploited by the creation of a male-sterile line, which has great potential for application in crop heterosis. Therefore, it is important to understand the molecular mechanisms of anther abortion induced by HT in wheat, which remain unclear at present. In this study, we performed phenotype improve language in the abstract and comparative transcriptome analysis of the male sterile anthers induced by HT in wheat. Compared with Normal anthers, the cytological analysis indicated that HT-induced male sterile anthers were smaller and had no starch accumulation in pollen grains, which is consistent with the results observed by scanning electron microscopy (SEM). The 9601 differentially expressed genes (DEGs) identified by transcriptome sequencing compared with the Normal anthers were noticeably involved in the following pathways: starch and sucrose metabolism, phosphatidylinositol (PI) signaling system, peroxidase activity and response to oxidative stress, and heme binding. In addition, TUNEL assays were performed and the results further confirmed the excessive accumulation of reactive oxygen species (ROS) in sterile anthers. Moreover, a total of 38 hub genes were obtained from the protein-protein interaction network analysis of these pathways, including genes, for example, heat shock protein 90 (HSP90), thioredoxin-like protein 1, peroxidase (POD), calreticulin, UDP glucose pyrophosphorylase (UGPase), sucrose synthase, phosphatidylinositol-4-phosphate 5-Kinase (PIP5K), cytochrome c, and Cystathionine beta-synthase X6-like (CBSX6-like). These findings provide insights for predicting the functions of the candidate genes, and the comprehensive analysis of our results is helpful for studying the abortive interaction mechanism induced by HT in wheat.

Microbial-Assisted Wheat Iron Biofortification Using Endophytic Bacillus altitudinis WR10.

Microbial-assisted biofortification attracted much attention recently due to its sustainable and eco-friendly nature for improving nutrient content in wheat. An endophytic strain Bacillus altitudinis WR10, which showed sophistical regulation of iron (Fe) homeostasis in wheat seedlings, inspired us to test its potential for enhancing Fe biofortification in wheat grain. In this study, assays in vitro indicated that WR10 has versatile plant growth-promoting (PGP) traits and bioinformatic analysis predicted its non-pathogenicity. Two inoculation methods, namely, seed soaking and soil spraying, with 107 cfu/ml WR10 cells were applied once before sowing of wheat (Triticum aestivum L. cv. Zhoumai 36) in the field. After wheat maturation, evaluation of yield and nutrients showed a significant increase in the mean number of kernels per spike (KPS) and the content of total nitrogen (N), potassium (K), and Fe in grains. At the grain filling stage, the abundance of Bacillus spp. and the content of N, K, and Fe in the root, the stem, and the leaf were also increased in nearly all tissues, except Fe in the stem and the leaf. Further correlation analysis revealed a positive relationship between the total abundance of Bacillus spp. and the content of N, K, and Fe in grains. Seed staining confirmed the enhanced accumulation of Fe, especially in the embryo and the endosperm. Finally, using a hydroponic coculture model, qPCR quantification indicated effective colonization, internalization, translocation, and replication of strain WR10 in wheat within 48 h. Collectively, strain WR10 assisted successful Fe biofortification in wheat in the field, laying a foundation for further large-scale investigation of its applicability and effectiveness.

The Sugar Transporter family in wheat (Triticum aestivum. L): genome-wide identification, classification, and expression profiling during stress in seedlings.

The sugar transporter protein (STP) plays a crucial role in regulating plant growth and stress tolerance. We performed genome-wide identification and expression analysis of the STP gene family to investigate the STPSs' potential roles in the growth of wheat seedlings under stress. Here, a total of 81 TaSTP genes containing the Sugar_tr conserved motif were identified within the wheat genome. Bioinformatic studies including phylogenetic tree, chromosome position, and tandem repeat were performed to analyze the identified genes. The 81 TaSTP genes can be classified into five main groups according to their structural and phylogenetic features, with several subgroups, which were located separately on chromosomes A, B, and D. Moreover, six gene clusters were formed with more than three genes each. The results of three comparative syntenic maps of wheat associated with three representative species suggested that STP genes have strong relationships in monocots. qRT-PCR analysis confirmed that most TaSTP genes displayed different expression profiles after seedlings were subjected to six days of different stress (10% PEG6000, 150 mM NaCl, and their combination, respectively), suggesting that these genes may be involved in regulating plant growth and stress tolerance. In conclusion, 81 TaSTP genes were identified and their expressions changed under stress, indicating TaSTP's potential roles in wheat growth monosaccharide distribution is regulated.

Endophytic Bacillus altitudinis WR10 alleviates Cu toxicity in wheat by augmenting reactive oxygen species scavenging and phenylpropanoid biosynthesis.

Soil copper (Cu) pollution severely stunts crops growth and limits sustainable agri-food production. Many microbes are widely used for remediation of polluted soil, including Cu pollution. In this study, the potential of an endophytic Bacillus altitudinis WR10 to protect wheat from Cu stress and the molecular mechanisms were investigated using hydroponic model. The Cu resistance assay showed B. altitudinis WR10 can resist up to 2 mM Cu and remove about 74% Cu in medium after 24 h of fermentation. Co-culture study demonstrated WR10 increased roots length and dry weight in wheat seedlings under 50 µM Cu. These results indicated that WR10 was a Cu-resistant strain and reduced Cu toxicity in wheat. Transcriptome data and biochemical tests of wheat roots indicated that WR10 alleviated Cu toxicity through enhancing peroxidases (PODs) gene expression and activity to remove excess hydrogen peroxide (H2O2) and down-regulating glutathione S-transferases (GSTs) to increase glutathione (GSH) level. Moreover, enrichment and pathway analysis indicated WR10 regulated the expression of genes involved in phenylpropanoid biosynthesis, which may improve phenolic acids accumulation for protecting plant cells from Cu toxicity. Overall, this study revealed that B. altitudinis WR10 alleviated Cu toxicity in wheat via augmenting reactive oxygen species scavenging and phenylpropanoid biosynthesis.

Complete Genome Sequence of Bacillus sp. strain WR11, an Endophyte Isolated from Wheat Root Providing Genomic Insights into Its Plant Growth-Promoting Effects.

Bacillus sp. strain WR11 isolated from the root of wheat (Triticum aestivum L.) possesses abiotic stress alleviating properties and produces several types of enzymes. However, its genomic information is lacking. The study described the complete genome sequence of the bacterium. The size of the genome was 4 202 080 base pairs that consisted of 4 405 genes in total. The G+C content of the circular genome was 43.53% and there were 4 170 coding genes, 114 pseudo genes, 30 ribosome RNAs, 86 tRNAs, and 5 ncRNAs, based on the Prokaryotic Genome Annotation Pipeline (PGAP). Many genes were related to the stress-alleviating properties and 124 genes existed in the CAZy database. The complete genome data of strain WR11 will provide valuable resources for genetic dissection of its plant growth-promoting function and symbiotic interaction with plant.

Trichoderma harzianum mitigates salt stress in cucumber via multiple responses.

Trichoderma harzianum T-soybean plays an important role in controlling soybean root rot disease. However, the mechanism by which it improves plant tolerance to salt stress is not clear. In this study, we investigated the possible mechanism of T-soybean in mitigating the damage caused by salt stress in Cucumis sativus L plants. Our results suggest that T-soybean improved salt tolerance of cucumber seedlings by affecting the antioxidant enzymes including peroxidase (POD) (EC 1.11.1.6), polyphenol oxidase (PPO) (EC 1.14.18.1), phenylalanine ammonia-lyase (PAL) (EC 4.3.1.5), catalase (CAT) (EC 1.11.1.6), superoxide dismutase (SOD) (EC 1.15.1.1), ascorbate peroxidase (APX) (EC 1.11.1.11), and glutathione reductase (GR) (EC 1.6.4.2), by increasing the levels of proline, soluble sugars, soluble protein, ascorbic acid (AsA) and chlorophyll as well as improving root activity. Treatment with T-soybean improved the ratio of glutathione (GSH)/oxidized glutathione (GSSG) and AsA/dehydroascorbate (DHA), and up-regulated the expression of CsAPX and CsGR genes involved in the AsA-GSH cycle. In addition, treatment with T-soybean increased the K+ content and K+/Na+ ratio while decreased the Na+ concentration and ethylene level. In summary, the improved salt tolerance of cucumber plants may be due to multiple mechanisms of T-soybean, such as the increase in reactive oxygen species (ROS) scavenging, as well as maintaining osmotic balance and metabolic homeostasis under salt stress.

Genome-Wide Identification and Characterization of Long Non-Coding RNA in Wheat Roots in Response to Ca2+ Channel Blocker.

It remains unclear whether plant lncRNAs are responsive to Ca2+-channel blocking. When using the Ca2+-channel blocker, LaCl3, to treat germinated wheat seeds for 24 h, we found that both root length and mitosis were inhibited in the LaCl3-treated groups. The effect of the Ca2+-channel blocker was verified in three ways: a [Ca2+]cyt decrease detected using Fluo-3/AM staining, a decrease in the Ca content measured using inductively coupled plasma mass spectrometry, and an inhibition of Ca2+ influx detected using Non-invasive Micro-test Technology. Genome-wide high throughput RNA-seq and bioinformatical methods were used to identify lncRNAs, and found 177 differentially expressed lncRNAs that might be in responsive to Ca2+-channel blocking. Among these, 108 were up-regulated and 69 were down-regulated. The validity of identified lncRNAs data from RNA-seq was verified using qPCR. GO and KEGG analysis indicated that a number of lncRNAs might be involved in diverse biological processes upon Ca2+-channel blocking. Further GO analysis showed that 23 lncRNAs might play roles as transcription factor (TF); Moreover, eight lncRNAs might participate in cell cycle regulation, and their relative expressions were detected using qPCR. This study also provides diverse data on wheat lncRNAs that can deepen our understanding of the function and regulatory mechanism of Ca2+-channel blocking in plants.

The complete mitochondrial genome of the Chinese rhesus macaques, Macaca mulatta lasiota.

In this study, the complete mitochondrial genome sequence of the Chinese rhesus macaques Macaca mulatta lasiota has been reported for the first time. The total length of the mitogenome was 16,561 bp. It contained the typical structure, including 13 protein-coding genes, 22 transfer RNA genes, 2 ribosomal RNA genes and 1 control region. The overall composition of the mitogenome was A (31.7%), G (12.8%), C (30.4%) and T (25.1%). The M. mulatta lasiota mitogenome had 21 tRNA genes folded into a typical clover-leaf secondary structure except for tRNA(Ser).