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

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