A membrane associated tandem kinase from wild emmer wheat confers broad-spectrum resistance to powdery mildew.

Crop wild relatives offer natural variations of disease resistance for crop improvement. Here, we report the isolation of broad-spectrum powdery mildew resistance gene Pm36, originated from wild emmer wheat, that encodes a tandem kinase with a transmembrane domain (WTK7-TM) through the combination of map-based cloning, PacBio SMRT long-read genome sequencing, mutagenesis, and transformation. Mutagenesis assay reveals that the two kinase domains and the transmembrane domain of WTK7-TM are critical for the powdery mildew resistance function. Consistently, in vitro phosphorylation assay shows that two kinase domains are indispensable for the kinase activity of WTK7-TM. Haplotype analysis uncovers that Pm36 is an orphan gene only present in a few wild emmer wheat, indicating its single ancient origin and potential contribution to the current wheat gene pool. Overall, our findings not only provide a powdery mildew resistance gene with great potential in wheat breeding but also sheds light into the mechanism underlying broad-spectrum resistance.

A combination of resistance genes confers high and durable resistance against stripe rust in wheat cultivar Lantian 26.

Lantian 26, a leading elite winter wheat cultivar in Gansu Province since its release in 2010, exhibits high resistance or immunization to stripe rust in adult-plant stage under a high disease pressure in Longnan (southeastern Gansu). Identifying the resistance genes in Lantian 26 could provide a basis for enhanced durability and high levels of resistance in wheat cultivars. Here, a segregating population was developed from a cross between a highly susceptible wheat cv. Mingxian 169 and the highly stripe rust-resistant cv. Lantian 26. The F2 and F2:3 progenies of the cross were inoculated with multiple prevalent virulent races of stripe rust for adult plant-stage resistance evaluation in two different environments. Exon sequence alignment analysis revealed that a stripe rust resistance gene on the 718.4-721.2 Mb region of chromosome 7BL, tentatively named as YrLT26, and a co-segregation STS marker GY17 was developed and validated using the F2:3 population and 103 wheat cultivars. The other two resistance genes, Yr9 and Yr30, were also identified in Lantian 26 using molecular markers. Therefore, the key to high and durable resistance to stripe rust at adult stage is the combination of Yr9, Yr30 and YrLT26 genes in Lantian 26. This could be a considerable strategy for improving the wheat cultivars with effective and durable resistance in the high-pressure region for stripe rust.

Lipopolysaccharide inhibits osteoblast formation and receptor activator of nuclear factor-κB ligand degradation via autophagy inhibition.

Lipopolysaccharide (LPS) and Receptor Activator of Nuclear Factor-κB Ligand (RANKL) are the two important factors causing bone loss, which is an important pathogenesis for osteoporosis. However, the relationship between LPS and RANKL is not yet clear. LPS can be involved in the weakened osteoblast formation as an autophagy regulator, and osteoblasts and their precursors are the source cells for RANKL production. Our study aimed to explore the relationship between autophagy changes and RANKL production during LPS-regulated osteoblasts. Our results showed that LPS inhibited autophagy (LC3 conversion and autophagosome formation) and enhanced the protein and mRNA expression of RANKL in MC3T3-E1 osteoblast precursor line. Autophagy upregulation with Rapamycin over BECN1 overexpression rescued LPS-inhibited osteoblast formation and -promoted RANKL protein production in MC3T3-E1 cells. In vivo experiments supported that damaged bone mass, bone microstructure, osteoblastic activity (ALP and P1NP production by ELISA assays) and enhanced RANKL production by LPS administration were partially rescued by Rapamycin application. In conclusion, LPS can inhibit autophagy in osteoblast precursors, thereby inhibiting osteoblast formation and RANKL autophagic degradation.

Modulation of high-quality internal multifoci based on modified three-dimensional Fourier transform.

We report an efficient method to generate arbitrary three-dimensional (3D) parallel multifoci inside a material. Taking into account the numerical aperture of the objective lens and the refractive index of the material, the Ewald cap was modified with a longer radius, then the whole 3D intensity distribution inside the material could be calculated using only a single Fourier transform (FT). By introducing the adaptive weight coefficient, the uniformity of the 3D multifoci improves from 81.3% to 98.9%. By adjusting the axial resolution of the Ewald cap, the uniformity of the axial multifoci improves from 85.9% to 99.7%. In the experiment, we have realized one-dimensional (1D), 2D, and 3D structures inside the fused silica, which are in excellent agreement with the simulation results. The experimental results of the "H-U-S-T" structure demonstrate that customized arbitrary intensity distribution inside the material can be realized.

Accuracy and digital screw path design of TiRobot-assisted pedicle screw placement for lumbar spondylolisthesis.

PURPOSE: To investigate lumbar spondylolisthesis screw placement assisted by TiRobot in terms of digital screw path design, accurate implementation, and accuracy evaluation method. METHODS: In this study, we enrolled 40 patients with lumbar spondylolisthesis between December 2020 and August 2021 who underwent spine surgery at the Affiliated Hospital of PuTian University. Pre-operative computed tomography position and screw path designation, intra-operative pedicle screw placement according to pre-operative planning, and post-operative evaluation of the accuracy of screw placement were performed. 3D coordinates of the entry and exit points before and after the operation were collected. The qualified points at different levels of accuracy were counted. The screw placement accuracy was based on the absolute difference using the Chi-squared test. RESULTS: In total, 194 screws were successfully implanted with no screws penetrating the cortex. The absolute difference of entry points X, Y, and Z coordinates before and after the operation was 0.425 ± 0.294 mm, 0.417 ± 0.310 mm, and 0.466 ± 0.327 mm, respectively. The corresponding values in terms of exit points were 0.702 ± 0.470 mm, 0.963 ± 0.595mm, and 0.983 ± 0.566 mm, respectively. No obvious differences in coordinates before and after the operation were observed with an entry point degree of accuracy of ≥ 1.2 mm and exit point degree of accuracy of ≥ 2.1 mm. Therefore, the real surgery was consistent with the design. CONCLUSIONS: TiRobot-assisted lumbar spondylolisthesis surgery achieved optimal path designation and precise surgery.

Φ-OTDR Signal Identification Method Based on Multimodal Fusion.

Distributed fiber optic sensing (DFS) systems are an effective method for long-distance pipeline safety inspections. Highly accurate vibration signal identification is crucial to DFS. In this paper, we propose an end-to-end high-accuracy fiber optic vibration signal detection and identification algorithm by extracting features from the time domain and frequency domain by a one-dimensional convolutional neural network and two-dimensional convolutional neural network, respectively, and introducing a self-attentive mechanism to fuse the features of multiple modes. First, the raw signal is segmented and normalized according to the statistical characteristics of the vibration signal combined with the distribution of noise. Then, the one-dimensional sequence of vibration signal and its two-dimensional image generated by short-time Fourier transform are input to the one-dimensional convolutional neural network and two-dimensional neural network, respectively, for automatic feature extraction, and the features are combined by long and short-time memory. Finally, the multimodal features generated from the time and frequency domains are fused by a multilayer TransformerEncoder structure with a multiheaded self-attentive mechanism and fed into a multilayer perceptron for classification. Experiments were conducted on an urban field database with complex noise and achieved 98.54% accuracy, which demonstrates the effectiveness of the proposed algorithm.

Coordinate-transformation iteration algorithm for phase modulation of arbitrary axial light distribution.

Axial light distribution modulation is widely applied in optical tweezers, hard-brittle material cutting, multilayer laser direct writing, etc. To generate arbitrary axial light distribution, the coordinate-transformation iteration (CTI) algorithm is presented. The CTI algorithm unifies equations in low and high numerical aperture (NA) scenarios, using the same iterative algorithm to produce phase computer-generated holograms. In a low NA scenario, twin-foci, flattop, and sin2 distributions have been achieved. In high NA scenarios, multirings, multifoci, and needle-like distributions have been realized in simulation with specific polarized incident beams. Since the CTI algorithm is inherently an efficient one-dimensional phase retrieval algorithm that is not limited by NA, this method has the potential to become a well-received solution for axial light distribution modulation.

Stealth dicing of 1-mm-thick glass with aberration-free axial multi-focus beams.

Laser stealth dicing can realize material separation with negligible surface damage, but severe aberrations in thick materials degrade processing quality. This Letter presents a nonlinear point-to-point transformation method combined with spherical aberration compensation to achieve aberration-free axial multi-focus beams. The focus peak intensity increases 7 times at a depth of 0.5 mm after spherical aberration compensation, and reaches 44 times at 3.5 mm. Spherical aberration compensation experiments showed that the width of the heat-affected zone remains almost unchanged at different depths inside the glass, and stealth dicing experiments for 1-mm-thick glass demonstrated that aberration-free 1-focus, 2-foci, and 3-foci stealth dicing can be successfully realized.

Full-endoscopic foraminoplasty for highly down-migrated lumbar disc herniation.

BACKGROUND AND STUDY AIMS: Multiple surgical approaches have been studied and accepted for the removal of highly downward migrated lumbar disc herniation (LDH). Here, we investigated the efficacy and safety of full-endoscopic foraminoplasty for highly downward migrated LDH. PATIENTS AND METHODS: Thirty-seven patients with highly down-migrated LDH treated by the full-endoscopic foraminoplasty between January 2018 and January 2020 were retrospectively investigated. Clinical parameters were evaluated preoperatively and 1, 6, and 12 months postoperatively, using pre- and post-operative Oswestry Disability Index (ODI) scores for functional improvement, visual analog scale (VAS) for leg and back pain, and modified MacNab criteria for patients satisfactory. RESULTS: Thirty-seven patients with highly downward migrated LDH were successfully removed via the transforaminal full-endoscopic discectomy. The average VAS back and leg pain scores were significantly reduced from 7.41 ± 1.17 and 8.68 ± 1.06 before operation to 3.14 ± 0.89 and 2.70 ± 0.46 at postoperative 1 month, and 1.76 ± 0.59 and 0.92 ± 0.28 at postoperative 12 months, respectively (P < 0.05). The average ODI scores were reduced from 92.86 ± 6.41 to 15.30 ± 4.43 at postoperative 1 month, and 9.81 ± 3.24 at postoperative 12 months (P < 0.05). Based on the modifed MacNab criteria, 36 out of 37 patients (97.30%) were rated as excellent or good outcomes. CONCLUSION: The full-endoscopic foraminoplasty can be used successfully for surgical removal of high grade down-migrated LDH, and it could serve as an efficient alternative technique for patients with highly downward migrated LDH.

Functional characterization of powdery mildew resistance gene MlIW172, a new Pm60 allele and its allelic variation in wild emmer wheat.

Wild emmer wheat (Triticum dicoccoides, WEW) is an immediate progenitor of both the cultivated tetraploid and hexaploid wheats and it harbors rich genetic diversity against powdery mildew caused by Blumeria graminis f. sp. tritici (Bgt). A powdery mildew resistance gene MlIW172 originated from WEW accession IW172 (G-797-M) is fine mapped in a 0.048 centimorgan (cM) genetic interval on 7AL, corresponding to a genomic region spanning 233 kb, 1 Mb and 800 kb in Chinese Spring, WEW Zavitan, and T. urartu G1812, respectively. MlIW172 encodes a typical NLR protein NLRIW172 and physically locates in an NBS-LRR gene cluster. NLRIW172 is subsequently identified as a new allele of Pm60, and its function is validated by EMS mutagenesis and transgenic complementation. Haplotype analysis of the Pm60 alleles reveals diversifications in sequence variation in the locus and presence and absence variations (PAV) in WEW populations. Four common single nucleotide variations (SNV) are detected between the Pm60 alleles from WEW and T. urartu, indicative of speciation divergence between the two different wheat progenitors. The newly identified Pm60 alleles and haplotypes in WEW are anticipated to be valuable for breeding powdery mildew resistance wheat cultivars via marker-assisted selection.

Fine mapping of powdery mildew resistance gene MlWE74 derived from wild emmer wheat (Triticum turgidum ssp. dicoccoides) in an NBS-LRR gene cluster.

KEY MESSAGE: Powdery mildew resistance gene MlWE74, originated from wild emmer wheat accession G-748-M, was mapped in an NBS-LRR gene cluster of chromosome 2BS. Wheat powdery mildew, caused by Blumeria graminis f. sp. tritici (Bgt), is a globally devastating disease. Wild emmer wheat (Triticum turgidum var. dicoccoides) is a valuable genetic resource for improving disease resistance in common wheat. A powdery mildew resistance gene was transferred to hexaploid wheat line WE74 from wild emmer accession G-748-M. Genetic analysis revealed that the powdery mildew resistance in WE74 is controlled by a single dominant gene, herein temporarily designated MlWE74. Bulked segregant analysis (BSA) and molecular mapping delimited MlWE74 to the terminal region of chromosome 2BS flanking by markers WGGBD412 and WGGBH346 within a genetic interval of 0.25 cM and corresponding to 799.9 kb genomic region in the Zavitan reference sequence. Sequence annotation revealed two phosphoglycerate mutase-like genes, an alpha/beta-hydrolases gene, and five NBS-LRR disease resistance genes that could serve as candidates for map-based cloning of MlWE74. The geographical location analysis indicated that MlWE74 is mainly distributed in Rosh Pinna and Amirim regions, in the northern part of Israel, where environmental conditions are favorable to the occurrence of powdery mildew. Moreover, the co-segregated marker WGGBD425 is helpful in marker-assisted transfer of MlWE74 into elite cultivars.

A CNL protein in wild emmer wheat confers powdery mildew resistance.

Powdery mildew, a fungal disease caused by Blumeria graminis f. sp. tritici (Bgt), has a serious impact on wheat production. Loss of resistance in cultivars prompts a continuing search for new sources of resistance. Wild emmer wheat (Triticum turgidum ssp. dicoccoides, WEW), the progenitor of both modern tetraploid and hexaploid wheats, harbors many powdery mildew resistance genes. We report here the positional cloning and functional characterization of Pm41, a powdery mildew resistance gene derived from WEW, which encodes a coiled-coil, nucleotide-binding site and leucine-rich repeat protein (CNL). Mutagenesis and stable genetic transformation confirmed the function of Pm41 against Bgt infection in wheat. We demonstrated that Pm41 was present at a very low frequency (1.81%) only in southern WEW populations. It was absent in other WEW populations, domesticated emmer, durum, and common wheat, suggesting that the ancestral Pm41 was restricted to its place of origin and was not incorporated into domesticated wheat. Our findings emphasize the importance of conservation and exploitation of the primary WEW gene pool, as a valuable resource for discovery of resistance genes for improvement of modern wheat cultivars.

A rare single nucleotide variant in Pm5e confers powdery mildew resistance in common wheat.

Powdery mildew poses severe threats to wheat production. The most sustainable way to control this disease is through planting resistant cultivars. We report the map-based cloning of the powdery mildew resistance allele Pm5e from a Chinese wheat landrace. We applied a two-step bulked segregant RNA sequencing (BSR-Seq) approach in developing tightly linked or co-segregating markers to Pm5e. The first BSR-Seq used phenotypically contrasting bulks of recombinant inbred lines (RILs) to identify Pm5e-linked markers. The second BSR-Seq utilized bulks of genetic recombinants screened from a fine-mapping population to precisely quantify the associated genomic variation in the mapping interval, and identified the Pm5e candidate genes. The function of Pm5e was validated by transgenic assay, loss-of-function mutants and haplotype association analysis. Pm5e encodes a nucleotide-binding domain leucine-rich-repeat-containing (NLR) protein. A rare nonsynonymous single nucleotide variant (SNV) within the C-terminal leucine rich repeat (LRR) domain is responsible for the gain of powdery mildew resistance function of Pm5e, an allele endemic to wheat landraces of Shaanxi province of China. Results from this study demonstrate the value of landraces in discovering useful genes for modern wheat breeding. The key SNV associated with powdery mildew resistance will be useful for marker-assisted selection of Pm5e in wheat breeding programs.

Identification and fine mapping of spot blotch (Bipolaris sorokiniana) resistance gene Sb4 in wheat.

KEY MESSAGE: A new spot blotch (Bipolaris sorokiniana) resistance gene Sb4 was mapped in a genomic interval of 1.34 Mb on wheat chromosome 4BL. Spot blotch, caused by Bipolaris sorokiniana, has emerged as a serious concern for cultivation of wheat in warmer and humid regions of the world, which results in substantial yield losses and descends with quality. In this study, we identified and mapped a spot blotch resistance gene, designated as Sb4, against B. sorokiniana in wheat. Bulked segregant RNA-Seq (BSR-Seq) analysis and single-nucleotide polymorphism mapping showed that Sb4 is located on the long arm of chromosome 4B. A genetic linkage map of Sb4 was constructed using an F4 mapping population developed from the cross between 'GY17' and 'Zhongyu1211,' and Sb4 was delimited in a 7.14-cM genetic region on 4BL between markers B6811 and B6901. Using the Chinese Spring reference sequences of chromosome arm 4BL, 13 new polymorphic markers were developed. Finally, Sb4 was mapped in a 1.19-cM genetic interval corresponding to a 1.34-Mb physical genomic region of Chinese Spring chromosome 4BL containing 21 predicted genes. This study provides a foundational step for further cloning of Sb4 using a map-based approach.

The Biosynthesis Mechanism Involving 2,3-Pentanedione and Aminoacetone Describes the Production of 2-Ethyl-3,5-dimethylpyrazine and 2-Ethyl-3,6-dimethylpyrazine by Bacillus subtilis.

2-Ethyl-3,5(3,6)-dimethylpyrazines (EDMPs) have a pleasant aroma of roasted cocoa or nuts with an extreme low odor threshold that have potential in industrial applications as food fragrances. The food fermentation process can accumulate EDMPs, and this might be the chance to study the biosynthesis mechanism of EDMPs under mild conditions for "natural" EDMPs' production. In this study, an EDMP-producing strain was isolated from baijiu fermentation. This strain was identified as Bacillus subtilis, a generally regarded as safe organism. After reasonable assumption and substrate addition and isotope-labeled experiments, we found that EDMPs are produced from l-threonine and d-glucose at environmental temperature and pressure. In addition, aminoacetone, the metabolite of l-threonine, and 2,3-pentanedione, the metabolite of l-threonine and d-glucose, are intermediates for the production of EDMPs. This study proposed and confirmed the biosynthesis pathway of EDMPs. It will be helpful for the industrial production of EDMPs and provides reference for the biosynthetic mechanism analysis of other valuable pyrazines.

Substitution of manure for chemical fertilizer affects soil microbial community diversity, structure and function in greenhouse vegetable production systems.

Soil microbial communities and enzyme activities together affect various ecosystem functions of soils. Fertilization, an important agricultural management practice, is known to modify soil microbial characteristics; however, inconsistent results have been reported. The aim of this research was to make a comparative study of the effects of different nitrogen (N) fertilizer rates and types (organic and inorganic) on soil physicochemical properties, enzyme activities and microbial attributes in a greenhouse vegetable production (GVP) system of Tianjin, China. Results showed that manure substitution of chemical fertilizer, especially at a higher substitution rate, improved soil physicochemical properties (higher soil organic C (SOC) and nutrient (available N and P) contents; lower bulk densities), promoted microbial growth (higher total phospholipid fatty acids and microbial biomass C contents) and activity (higher soil hydrolase activities). Manure application induced a higher fungi/bacteria ratio due to a lower response in bacterial than fungal growth. Also, manure application greatly increased bacterial stress indices, as well as microbial communities and functional diversity. The principal component analysis showed that the impact of manure on microbial communities and enzyme activities were more significant than those of chemical fertilizer. Furthermore, redundancy analysis indicated that SOC and total N strongly influenced the microbial composition, while SOC and ammonium-N strongly influenced the microbial activity. In conclusion, manure substitution of inorganic fertilizer, especially at a higher substitution rate, was more efficient for improving soil quality and biological functions.

A rare gain of function mutation in a wheat tandem kinase confers resistance to powdery mildew.

Powdery mildew, caused by Blumeria graminis f. sp. tritici (Bgt), is one of the most destructive diseases that pose a great threat to wheat production. Wheat landraces represent a rich source of powdery mildew resistance. Here, we report the map-based cloning of powdery mildew resistance gene Pm24 from Chinese wheat landrace Hulutou. It encodes a tandem kinase protein (TKP) with putative kinase-pseudokinase domains, designated WHEAT TANDEM KINASE 3 (WTK3). The resistance function of Pm24 was validated by transgenic assay, independent mutants, and allelic association analyses. Haplotype analysis revealed that a rare 6-bp natural deletion of lysine-glycine codons, endemic to wheat landraces of Shaanxi Province, China, in the kinase I domain (Kin I) of WTK3 is critical for the resistance function. Transgenic assay of WTK3 chimeric variants revealed that only the specific two amino acid deletion, rather than any of the single or more amino acid deletions, in the Kin I of WTK3 is responsible for gaining the resistance function of WTK3 against the Bgt fungus.