Nucleotide diversity and molecular evolution of the WAG-2 gene in common wheat (Triticum aestivum L) and its relatives

In this work, we examined the genetic diversity and evolution of the WAG-2 gene based on new WAG-2 alleles isolated from wheat and its relatives. Only single nucleotide polymorphisms (SNP) and no insertions and deletions (indels) were found in exon sequences of WAG-2 from different species. More SNPs and indels occurred in introns than in exons. For exons, exons+introns and introns, the nucleotide polymorphism π decreased from diploid and tetraploid genotypes to hexaploid genotypes. This finding indicated that the diversity of WAG-2 in diploids was greater than in hexaploids because of the strong selection pressure on the latter. All dn/ds ratios were < 1.0, indicating that WAG-2 belongs to a conserved gene affected by negative selection. Thirty-nine of the 57 particular SNPs and eight of the 10 indels were detected in diploid species. The degree of divergence in intron length among WAG-2 clones and phylogenetic tree topology suggested the existence of three homoeologs in the A, B or D genome of common wheat. Wheat AG-like genes were divided into WAG-1 and WAG-2 clades. The latter clade contained WAG-2, OsMADS3 and ZMM2 genes, indicating functional homoeology among them

Cloning and characterization of endo-β-1,4-glucanase genes in the common wheat line three pistils

In this work, we report the cloning and characterization of endo-β-1,4-glucanase (EGase) genes (TaEG) in the common wheat line three pistils. Three TaEG homoeologous genes (TaEG-4A, TaEG-4B and TaEG-4D) were isolated and found to be located on chromosomes 4AL, 4BS and 4DS, respectively. The three genes showed high conservation of their coding nucleotide sequences and 3 untranslated region. The putative TaEG protein had a molecular mass of 69 kDa, a theoretical pI of 9.39 and a transmembrane domain of 74-96 amino acids in the N-terminus that anchored the protein to the membrane. The genome sequences of TaEG-4A, TaEG-4B and TaEG-4D contained six exons and five introns. All of the introns, except for intron IV, varied in length and sequence composition. Phylogenetic analysis revealed that TaEG was most closely related to rice (Oryza sativa) OsGLU1. The TaEG transcript levels increased significantly during the subsidiary pistil primordium differentiation phase (spike size ~7-10 mm) in Chuanmai 28 TP (CM28TP). These data provide a basis for future research into the function of TaEG and offer insights into the molecular mechanism of the three pistils mutation in wheat

Development of a high-density linkage map and mapping of the three-pistil gene (Pis1) in wheat using GBS markers

Abstract Background The wheat mutant line three-pistil (TP) exhibits three pistils per floret. As TP normally has two or three seeds in each of the florets on the same spike, there is the possibility of increasing the number of grains per spike. Therefore, TP is a highly valuable mutant for breeding and for the study of floral development in wheat. To map the three-pistil gene (Pis1), genotyping-by-sequencing single-nucleotide polymorphism (GBS-SNP) data from an F2 mapping population (CM28 × CM28TP) was used to construct a genetic map that is of significant value. Results In the present study, a high-density genetic map of wheat containing 2917 GBS-SNP markers was constructed. Twenty-one linkage groups were resolved, with a total length of 2371.40 cM. The individual chromosomes range from 2.64 cM to 454.55 cM with an average marker density of 0.81 cM. The Pis1 gene was mapped using this high-resolution map, and two flanking SNP markers tightly linked to the gene, M70 and M71, were identified. The Pis1 is 3.00 cM from M70 and 1.10 cM from M71. In bread wheat genome, M70 and M71 were found to delimit a physical distance of 3.40 Mb, which encompasses 127 protein-coding genes. To validate the GBS-generated genotypic data and to eliminate missing marker data in the Pis1 region, five Kompetitive Allele-Specific PCR (KASP) assays were designed from corresponding GBS sequences, which harbor SNPs that surround Pis1. Three KASP-SNP markers, KM70, KM71, and KM75, were remapped to the Pis1 gene region. Conclusions This work not only lays the foundation for the map-based cloning of Pis1 but can also serve as a valuable tool for studying marker-trait association of important traits and marker-assisted breeding in wheat

Mitigation of Lost Circulation in Oil-Based Drilling Fluids Using Oil Absorbent Polymers

In order to mitigate the loss circulation of oil-based drilling fluids (OBDFs), an oil-absorbent polymer (OAP) composed by methylmethacrylate (MMA), butyl acrylate (BA), and hexadecyl methacrylate (HMA) was synthesized by suspension polymerization and characterized by Fourier transform infrared spectroscopy (FT-IR), thermogravimetric analysis (TGA) and scanning electronic microscopy (SEM). The oil-absorptive capacity of OAP under different solvents was measured as the function of temperature and time. The effect of the OAP on the rheological and filtration properties of OBDFs was initially evaluated, and then the sealing property of OAP particles as lost circulation materials (LCMs) was examined by a high-temperature and high-pressure (HTHP) filtration test, a sand bed filtration test, a permeable plugging test, and a fracture sealing testing. The test results indicated that the addition of OAP had relatively little influence on the rheological properties of OBDF at content lower than 1.5 w/v % but increased the fluid viscosity remarkably at content higher than 3 w/v %. It could reduce the HTHP filtration and improve the sealing capacity of OBDF significantly. In the sealing treatment, after addition into the OBDF, the OAP particles could absorb oil accompanied with volume enlargement, which led to the increase of the fluid viscosity and slowing down of the fluid loss speed. The swelled and deformable OAP particles could be squeezed into the micro-fractures with self-adoption and seal the loss channel. More important, fluid loss was dramatically reduced when OAP particles were combined with other conventional LCMs by a synergistic effect

Effect of Amphiphilic Polymer/Nano-Silica Composite on Shale Stability for Water-Based Muds

Research on using nanotechnology to solve shale instability problems in drilling engineering has been increasing. The combination of amphiphilic polymer and silica nanoparticles may be a new way to improve shale stability. Herein, an amphiphilic polymer/nano-silica composite (poly(styrene-methyl methacrylate-acrylamide)/nano-SiO2) was introduced as a novel shale stabilizer SMA/SiO2 for water-based muds, which possessed the advantages of both physical plugging and chemical inhibition during the drilling operations. The SMA/SiO2 was prepared and characterized by Fourier transform infrared spectra (FT-IR), nuclear magnetic resonance (1H-NMR), transmission electron microscope (TEM), particle size distribution (PSD) and thermogravimetric analysis (TGA) experiments, which confirmed that SMA/SiO2 was regularly spherical with nano-scale and showed good high-temperature resistance. To evaluate the plugging capacity of SMA/SiO2, the pressure transmission test and BET analysis were applied. The results indicated SMA/SiO2 was capable of effectively plugging the pores and fractures in shale. To evaluate the hydration inhibition capacity of SMA/SiO2, the rolling dispersion experiment and contact angle test were adopted. The results demonstrated that SMA/SiO2 could reduce the tendency of shale hydration, which was better than potassium chloride (KCl) and polymeric alcohol (JHC). In addition, SMA/SiO2 only created slight variations on the rheological parameters of the water-based muds (WBMs) and showed a significant filtration control performance. Due to the outstanding performance of physical plugging and chemical inhibition, SMA/SiO2 was expected to be a novel shale stabilizer to solve shale instability problems

High-density genetic map construction and mapping of the homologous transformation sterility gene (hts) in wheat using GBS markers

Abstract Background Homologous transformation sterility-1 (HTS-1) is a novel wheat mutant that exhibits pistillody, the transformation of stamens into pistils or pistil-like structures. More extreme phenotypes of this mutation can have six pistils or pistil-like structures without any stamens in a floret. Thus, HTS-1 is highly valuable for studies of wheat hybrid breeding and flower development. Previous studies have shown that two major genes (Pis1 and hts) control pistillody in HTS-1. The Pis1 gene controls the three-pistil trait in the three-pistil wheat mutant and has been mapped on chromosome 2D, but the hts gene has not been mapped or identified. To do so, we crossed HTS-1 with CM28TP (three-pistil mutant) and constructed a high-density linkage map with the F2 population (200 individuals). Results The map covered 2779.96 cM, and the genetic distance per chromosome ranged from 37.59 cM to 318.95 cM. The average distance between markers was 1.04 cM. We then mapped hts between GBS-SNP markers 4A_109 and 4A_119, separated by 2.0 cM and 5.2 Mb. To find the candidate genes, the hts region was enlarged to 7.2 Mb, encompassing 752 protein-coding genes. We identified TaWin1 as a possible candidate gene after comparing the 752 genes with 206 common differentially expressed genes between pistillody stamens (PS) versus normal stamens (S) and pistils (P) versus S. Real-time PCR indicated that TaWin1 was highly expressed in HTS-1 during the pistil-and-stamen-differentiating stage, at levels approximately 120 times greater than those in CM28TP. Further analysis indicated that TaWin1 was mainly expressed in HTS-1 PS, supporting its status as a candidate gene of hts. Thus, TaWin1 overexpression probably leads to the transformation of stamens into pistils in wheat. Conclusions The results of this study provide a foundation for further research on stamen and pistil development, with implications for wheat-hybrid breeding programs

Novel fluorescent sequence-related amplified polymorphism(FSRAP) markers for the construction of a genetic linkage map of wheat(Triticum aestivum L.)

Novel fluorescent sequence-related amplified polymorphism (FSRAP) markers were developed based on the SRAP molecular marker. Then, the FSRAP markers were used to construct the genetic map of a wheat (Triticum aestivumL.) recombinant inbred line population derived from a Chuanmai 42×Chuannong 16 cross. Reproducibility and polymorphism tests indicated that the FSRAP markers have repeatability and better reflect the polymorphism of wheat varieties compared with SRAP markers. A total of 430 polymorphic loci between Chuanmai 42 and Chuannong 16 were detected with 189 FSRAP primer combinations. A total of 281 FSARP markers and 39 SSR markers re classified into 20 linkage groups. The maps spanned a total length of 2499.3cM with an average distance of 7.81cM between markers. A total of 201 markers were mapped on the B genome and covered a distance of 1013cM. On the A genome, 84 markers were mapped and covered a distance of 849.6cM. On the D genome, however, only 35 markers were mapped and covered a distance of 636.7cM. No FSRAP markers were distributed on the 7D chromosome. The results of the present study revealed that the novel FSRAP markers can be used to generate dense, uniform genetic maps of wheat