The most prevalent form of the Rubisco enzyme is a complex of eight catalytic large subunits (RbcL) and eight regulatory small subunits (RbcS). Rubisco biogenesis depends on the assistance by specific molecular chaperones. The assembly chaperone RbcX stabilizes the RbcL subunits after folding by chaperonin and mediates their assembly to the RbcL8 core complex, from which RbcX is displaced by RbcS to form active holoenzyme. Two isoforms of RbcX are found in eukaryotes, RbcX-I, which is more closely related to cyanobacterial RbcX, and the more distant RbcX-II. The green algae Chlamydomonas reinhardtii contains only RbcX-II isoforms, CrRbcX-IIa and CrRbcX-IIb. Here we solved the crystal structure of CrRbcX-IIa and show that it forms an arc-shaped dimer with a central hydrophobic cleft for binding the C-terminal sequence of RbcL. Like other RbcX proteins, CrRbcX-IIa supports the assembly of cyanobacterial Rubisco in vitro, albeit with reduced activity relative to cyanobacterial RbcX-I. Structural analysis of a fusion protein of CrRbcX-IIa and the C-terminal peptide of RbcL suggests that the peptide binding mode of RbcX-II may differ from that of cyanobacterial RbcX. RbcX homologs appear to have adapted to their cognate Rubisco clients as a result of co-evolution.
Abstract Substantial efforts are being made to optimize the CRISPR/Cas9 system for precision crop breeding. The avoidance of transgene integration and reduction of off-target mutations are the most important targets for optimization. Here, we describe an efficient genome editing method for bread wheat using CRISPR/Cas9 ribonucleoproteins (RNPs). Starting from RNP preparation, the whole protocol takes only seven to nine weeks, with four to five independent mutants produced from 100 immature wheat embryos. Deep sequencing reveals that the chance of off-target mutations in wheat cells is much lower in RNP mediated genome editing than in editing with CRISPR/Cas9 DNA. Consistent with this finding, no off-target mutations are detected in the mutant plants. Because no foreign DNA is used in CRISPR/Cas9 RNP mediated genome editing, the mutants obtained are completely transgene free. This method may be widely applicable for producing genome edited crop plants and has a good prospect of being commercialized.
Phosphoglucose isomerase (PGI) catalyzes the interconversion between glucose-6-phosphate (G6P) and fructose-6-phosphate (F6P), thereby regulating sucrose synthesis in plant cells. In general, plants contain a pair of PGI isozymes located in two distinct compartments of the cell (cytosol and plastid) with differences in both the primary structure and the higher-order structure. Previously, we showed that the activity of cytosolic PGI (PGIc) is more robust (activity, thermal stability, substrate turnover rate, etc.) than that of the plastid counterpart (PGIp) in multiple organisms, including wheat, rice, and Arabidopsis . The crystal structures of apoTaPGIc (an isotype cytosol PGIc in Triticum aestivum ), TaPGIc-G6P complex, and apoTaPGIp (an isotype plastid PGIp in Triticum aestivum ) were first solved in higher plants, especially in crops. In this study, we detailed the structural characteristics related to the biochemical properties and functions of TaPGIs in different plant organelles. We found that the C-terminal domains (CTDs) of TaPGIc and TaPGIp are very different, which affects the stability of the dimerized enzyme, and that Lys213 TaPGIc /Lys193 TaPGIp and its surrounding residues at the binding pocket gateway may participate in the entrance and exit of substrates. Our findings provide a good example illuminating the evolution of proteins from primary to higher structures as a result of physical barriers and adaptation to the biochemical environment.
Natural gas in the Ludong-Wucaiwan area is characterized by typical source-controlled reservoirs. Combining outcrop data, well data and seismic data, this article makes a systematic research and evaluation on the abundance, type, and maturity characteristics of the Carboniferous source rocks in the Ludong-Wucaiwan area. The Carboniferous source rocks are dominantly dark grey and grey mudstones, secondarily tuffs, carbonaceous mudstones, silty mudstones and coals. The source rock abundance of the Upper Carboniferous Batamayineishan Formation (C2b) is higher than that of the Lower Carboniferous Dishuiquan Formation (C1d), while the kerogen type of the Lower Carboniferous source rock is better than that of the Upper Carboniferous. The maturity center of the Carboniferous source rocks is located in the west part of Dinan uplift to the Wucaiwan depression area, with maturity decreased towards south and north. The distributions of the Carboniferous source rocks and volcanic rocks are complementary. The source rocks are distributed in zonal shape in east-west direction, and are most developed in the west part of the Dinan uplift and its southern area. : 陆东—五彩湾地区天然气具有典型的源控成藏特征,将露头资料、钻井资料和地震资料相结合,对陆东—五彩湾地区石炭系烃源岩的丰度、类型、成熟度特征进行了系统研究和评价。研究区石炭系烃源岩以深灰色泥岩和灰色泥岩为主,其次为凝灰岩、炭质泥岩、粉砂质泥岩和煤;上石炭统巴塔玛依内山组(C2b)烃源岩丰度高于下石炭统滴水泉组(C1d),下石炭统烃源岩的类型好于上石炭统;石炭系烃源岩成熟度高值区分布在滴南凸起西段—五彩湾凹陷一带,向南、北方向石炭系烃源岩的成熟度降低;石炭系烃源岩与火山岩的分布具有互补性,烃源岩呈东西向带状分布,烃源岩最发育区位于滴南凸起西段及其南部。图8表2参15
Abstract 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.
Several oil and gas fields have been found in which oil and gas were mainly derived from the Jurassic coaly source rocks in the Junggar Basin, northwest China. Pyrolysis experiments were performed on two coals (J23C1 and FM1C2) and one type III kerogen of mudstone (Di9S1) from Jurassic strata in the basin at two heating rates of 20 and 2 °C/h in confined systems (gold capsules). Hydrogen indices and H/C atomic ratios of the three samples J23C1, FM1C2, and Di9S1 are 83, 197, and 226 mg/g TOC, and 0.70, 0.86, and 1.01, respectively. The measured maximum oil yields for the three samples are 59.37, 175.75, and 80.75 mg/g TOC, respectively, inconsistent with hydrogen indices and H/C atomic ratios. However, the measured maximum gas yields (∑C1–5) for the three samples are 90.69, 157.24, and 198.15 mg/g TOC, respectively, consistent with hydrogen indices and H/C atomic ratios. This result is interpreted by kerogen Di9S1 containing mainly crossed alkane moieties with both terminals attached to aromatic rings while coals J23C1 and FM1C2 contain mainly alkane moieties with only one terminal attached to an aromatic ring based on kerogen 13C NMR spectra and the oil yield relative to gas yield and compositions of liquid components produced in confined pyrolysis. The crossed alkane moieties were hardly released as liquid alkanes but likely further cracked into gaseous components during pyrolysis. Jurassic strata contain some effective oil source rocks which produced enough amount of oil required for oil expulsion and formation of commercial oil reservoirs in oil generative window (Ro 0.6–1.35%). The amounts of gaseous hydrocarbons generated from the Jurassic coaly source rocks are generally low in oil generative window due to low transformation ratios. Elevated maturity (Ro > 1.35%) is a critical controlling factor to the Jurassic coaly source rocks generating sufficient gaseous hydrocarbons and forming commercial gas reservoirs.
Vesicle-inducing protein in plastids 1 (Vipp1) is thought to play an important role both in thylakoid biogenesis and chloroplast envelope maintenance during stress. Vipp1 is conserved in photosynthetic organisms and forms a high homo-oligomer complex structure that may help sustain the membrane integrity of chloroplasts. This study cloned two novel VIPP1 genes from Triticum urartu and named them TuVipp1 and TuVipp2. Both proteins shared high identity with the homologous proteins AtVipp1 and CrVipp1. TuVipp1 and TuVipp2 were expressed in various organs of common wheat, and both genes were induced by light and various stress treatments. Purified TuVipp1 and TuVipp2 proteins showed secondary and advanced structures similar to those of the homologous proteins. Similar to AtVipp1, TuVipp1 is a chloroplast target protein. Additionally, TuVipp1 was able to rescue the phenotypes of pale leaves, lethality, and disordered chloroplast structures of AtVipp1 (-/-) mutant lines. Collectively, our data demonstrate that TuVipp1 and TuVipp2 are functional proteins in chloroplasts in wheat and may be critical for maintaining the chloroplast envelope under stress and membrane biogenesis upon photosynthesis.
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