The EXO70 gene family is involved in different biological processes in plants, ranging from plant polar growth to plant immunity. To date, analysis of the EXO70 gene family has been limited in Triticeae species, e.g., hexaploidy Triticum aestivum and its ancestral/related species. By in silico analysis of multiple Triticeae sequence databases, a total of 200 EXO70 members were identified. By homologue cloning approaches, 15 full-length cDNA of EXO70s were cloned from diploid Haynaldia villosa. Phylogenetic relationship analysis of 215 EXO70 members classified them into three groups (EXO70.1, EXO70.2, and EXO70.3) and nine subgroups (EXO70A to EXO70I). The distribution of most EXO70 genes among different species/sub-genomes were collinear, implying their orthologous relationship. The EXO70A subgroup has the most introns (at least five introns), while the remaining seven subgroups have only one intron on average. The expression profiling of EXO70 genes from wheat revealed that 40 wheat EXO70 genes were expressed in at least one tissue (leaf, stem, or root), of which 25 wheat EXO70 genes were in response to at least one biotic stress (stripe rust or powdery mildew) or abiotic stress (drought or heat). Subcellular localization analysis showed that ten EXO70-V proteins had distinct plasma membrane localization, EXO70I1-V showed a distinctive spotted pattern on the membrane. The 15 EXO70-V genes were differentially expressed in three tissue. Apart from EXO70D2-V, the remaining EXO70-V genes were in response to at least one stress (flg22, chitin, powdery mildew, drought, NaCl, heat, or cold) or phytohormones (salicylic acid, methyl jasmonate, ethephon, or abscisic acid) and hydrogen peroxide treatments. This research provides a genome-wide glimpse of the Triticeae EXO70 gene family and those up- or downregulated genes require further validation of their biological roles in response to biotic/abiotic stresses.
BackgroundPowdery mildew, caused by Blumeria graminearum f.sp. tritici (Bgt), is one of the most severe fungal diseases of wheat. The exploration and utilization of new gene resources is the most effective approach for the powdery mildew control.ResultsWe report the cloning and functional analysis of two wheat LRR-RLKs from T. aestivum c.v. Prins- T. timopheevii introgression line IGV1-465, named TaRLK1 and TaRLK2, which play positive roles in regulating powdery mildew resistance in wheat. The two LRR-RLKs contain an ORF of 3,045 nucleotides, encoding a peptide of 1014 amino acids, with seven amino acids difference. Their predicted proteins possess a signal peptide, several LRRs, a trans-membrane domain, and a Ser/Thr protein kinase domain. In response to Bgt infection, the TaRLK1/2 expression is up-regulated in a developmental-stage-dependent manner. Single-cell transient over-expression and gene-silencing assays indicate that both genes positively regulate the resistance to mixed Bgt inoculums. Transgenic lines over-expressing TaRLK1 or TaRLK2 in a moderate powdery mildew susceptible wheat variety Yangmai 158 led to significantly enhanced powdery mildew resistance. Exogenous applied salicylic acid (SA) or hydrogen peroxide (H2O2) induced the expression of both genes, and H2O2 had a higher accumulation at the Bgt penetration sites in RLK over-expression transgenic plants, suggesting a possible involvement of SA and altered ROS homeostasis in the defense response to Bgt infection. The two LRR-RLKs are located in the long arm of wheat chromosome 2B, in which the powdery mildew resistance gene Pm6 is located, but in different regions.ConclusionsTwo members of TaRLK family were cloned from IGV1-465. TaRLK1 and TaRLK2 contribute to powdery mildew resistance of wheat, providing new resistance gene resources for wheat breeding.Electronic supplementary materialThe online version of this article (doi:10.1186/s12870-016-0713-8) contains supplementary material, which is available to authorized users.
BackgroundFull-percutaneous endoscopic lumbar discectomy (F-PELD) is a popular operation for the treatment of lumbar disc herniation (LDH). Some studies have reported that F-PELD in day surgery mode produced favorable outcomes for LDH. At the same time, minimally invasive spinal surgery following enhanced recovery after surgery (ERAS) presents a rising trend in recent years, but few studies reported whether F-PELD will produce better outcomes in the day surgery (DS) mode combined with ERAS.ObjectiveTo analyze whether F-PELD in day surgery mode following ERAS can produce better clinical outcomes than in traditional surgery mode.MethodsThe patients who underwent F-PELD between January 2019 and October 2020 were retrospectively analyzed, and the patients who met the inclusive criteria were followed up. The patients were divided into day surgery (DS) group (n = 152) that combined with ERAS and traditional surgery (TS) group (n = 123) without ERAS. The length of hospital stays (LOS), visual analogue scale (VAS), and Oswestry Disability Index (ODI) of two groups were compared before surgery, immediately after surgery, one month after surgery, and one year after surgery.ResultsA total of 298 patients who underwent F-PELD were reviewed. 290 patients were included in the study and followed up, and 275 patients who had completed the follow-up were available for analysis. There were no statistically significant differences between the two groups in terms of age, gender, preoperative VAS, and ODI. There were significant statistical differences in the VAS and ODI immediately after surgery (VAS for back pain: DS group 1.4 ± 1.1, TS group 2.0 ± 1.2, p < 0.001; VAS for leg pain: DS group 0.8 ± 0.8, TS group 1.1 ± 1.1, p = 0.010; ODI: DS group 5.8 ± 4.3, TS group 7.6 ± 7.4, p = 0.010) and one month after surgery (VAS for back pain: DS group 0.8 ± 0.9, TS group 1.1 ± 1.0, p = 0.035; ODI: DS group 3.2 ± 3.5, TS group 4.5 ± 6.5, p = 0.036). At one year after surgery, the VAS (back pain: DS group 0.3 ± 0.6, TS group 0.3 ± 0.7, p = 0.798; leg pain: DS group 0.2 ± 0.4, TS group 0.1 ± 0.4, p = 0.485) and ODI (DS group 0.8 ± 1.2, TS group 0.7 ± 1.7, p = 0.729) were further improved, but no statistically significant difference was observed between two groups. LOS of DS group (1.38 ± 0.49 days) was significantly shorter than the TS group (5.83 ± 2.24 days, p < 0.001), and some postoperative complications occurred in the TS group, including throat discomfort (n = 5, 4.1%), discomfort after catheterization (n = 7, 5.7%), abdominal distention (n = 3, 2.4%), and nausea (n = 5, 4.1%). None of the above complications resulted in serious consequences.ConclusionThe F-PELD in day surgery mode following ERAS produced a better short-term clinical effect and reduced the LOS, which is worthy of promotion.
Background: Short arm of chromosome 6V (6VS) of Haynaldia villosa has been used in wheat breeding programs to introduce Pm21 resistance gene against powdery mildew and some other genes. Results: In this work, 6VS was isolated from a wheat ( Triticum aestivum ) - 6VS telosome addition line by flow cytometric sorting and sequenced by illumina technology. The assembly length was 230.39 Mb with contig N50 of 9,788 bp. The sequence annotation identified 3,276 high confidence genes supported by RNA sequencing data, representing about 2.3% of the chromosome arm sequence; repetitive elements accounted for 74.91% of the arm sequence. Sequences homologous to 6VS genes were identified on short arms of chromosomes 6A of T. urartu , 6D of Aegilops tauschii , 6A and 6B of T. dicoccoides , 6A, 6B and 6D of T. aestivum and 6H of Hordeum vulgare , revealing synteny relationships among these chromosome arms. Based on differences in intron size between the homologous genes on 6VS and 6AS/6BS/6DS of T. aestivum , 222 primer pairs were designed. Out of them, 120 amplified 6VS-specific products and are suitable as intron-target (IT) markers to trace the 6VS chromatin introduced into wheat. Conclusions: The results obtained and markers developed in this work will facilitate introduction of important genes to common wheat from its wild relative, while reducing the presence of unfavorable genes due to linkage drag.
Background Haynaldia villosa (H. villosa) has been recognized as a species potentially useful for wheat improvement. The availability of its genomic sequences will boost its research and application.ResultsIn this work, the short arm of H. villosa chromosome 4V (4VS) was sorted by flow cytometry and sequenced using Illumina platform. About 170.6 Mb assembled sequences were obtained. Further analysis showed that repetitive elements accounted for about 64.6% of 4VS, while the coding fraction, which is corresponding to 1977 annotated genes, represented 1.5% of the arm. The syntenic regions of the 4VS were searched and identified on wheat group 4 chromosomes 4AL, 4BS, 4DS, Brachypodium chromosomes 1 and 4, rice chromosomes 3 and 11, and sorghum chromosomes 1, 5 and 8. Based on genome-zipper analysis, a virtual gene order comprising 735 gene loci on 4VS genome was built by referring to the Brachypodium genome, which was relatively consistent with the scaffold order determined for Ae. tauschii chromosome 4D. The homologous alleles of several cloned genes on wheat group 4 chromosomes including Rht-1 gene were identified.ConclusionsThe sequences provided valuable information for mapping and positional-cloning genes located on 4VS, such as the wheat yellow mosaic virus resistance gene Wss1. The work on 4VS provided detailed insights into the genome of H. villosa, and may also serve as a model for sequencing the remaining parts of H. villosa genome.Electronic supplementary materialThe online version of this article (10.1186/s12864-017-4211-7) contains supplementary material, which is available to authorized users.
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