BackgroundCellulosic biofuels are sustainable compared to fossil fuels. However, inhibitors, such as acetic acid generated during lignocellulose pretreatment and hydrolysis, would significantly inhibit microbial fermentation efficiency. Microbial mutants able to tolerate high concentration of acetic acid are needed urgently to alleviate this inhibition.ResultsZymomonas mobilis mutants AQ8-1 and AC8-9 with enhanced tolerance against acetic acid were generated via a multiplex atmospheric and room temperature plasma (mARTP) mutagenesis. The growth and ethanol productivity of AQ8-1 and AC8-9 were both improved in the presence of 5.0–8.0 g/L acetic acid. Ethanol yield reached 84% of theoretical value in the presence of 8.0 g/L acetic acid (~ pH 4.0). Furthermore, a mutant tolerant to pH 3.5, named PH1-29, was generated via the third round of ARTP mutagenesis. PH1-29 showed enhanced growth and ethanol production under both sterilized/unsterilized conditions at pH 4.0 or 3.5. Intracellular NAD levels revealed that mARTP mutants could modulate NADH/NAD+ ratio to respond to acetic acid and low pH stresses. Moreover, genomic re-sequencing revealed that eleven single nucleic variations (SNVs) were likely related to acetic acid and low pH tolerance. Most SNVs were targeted in regions between genes ZMO0952 and ZMO0956, ZMO0152 and ZMO0153, and ZMO0373 and ZMO0374.ConclusionsThe multiplex mutagenesis strategy mARTP was efficient for enhancing the tolerance in Z. mobilis. The ARTP mutants generated in this study could serve as potential cellulosic ethanol producers.Electronic supplementary materialThe online version of this article (10.1186/s13068-018-1348-9) contains supplementary material, which is available to authorized users.
Prostaglandins E2 (PGE2) has been shown to enhance IgE production by B cells in vitro. The physiological and pathological relevance of this phenomenon and the underlying molecular mechanism, however, remain to be elucidated. B cells from wild type and EP2-deficient mice were compared in culture for their responses to PGE2 in terms of IgE class switching and production. Ovalbumin (OVA)-induced asthma models were used to evaluate the impact of EP2-deficiency on IgE responses and the development of asthma. PGE2 promoted IgE class switching, generation of IgE+ cells and secretion of IgE by B cells stimulated with LPS+IL4. These effects were much attenuated as a consequence of EP2 deficiency. Consistent with the in vitro data, EP2-deficient mice showed a markedly suppressed IgE antibody response and developed less pronounced airway inflammation in the OVA-induced asthma model. Mechanistic studies demonstrated that PGE2, in an EP2-depedent manner, enhanced STAT6 activation induced by IL-4, thereby promoting the expression of IgE germline and post switch transcripts and the transcription of activation-induced cytidine deaminase (AID). Collectively, these data support an important regulatory role of the PGE2-EP2-STAT6 signaling pathway in IgE response and allergic diseases.
Salinity is one of the major abiotic stresses limiting rice production worldwide. Understanding the genetic basis of salinity tolerance is key for rice breeding. In this study, a recombinant inbred line (RIL) population derived from a super hybrid rice Liang-You-Pei-Jiu (LYP9) parents 93-11 and PA64s, exhibited variation in phenotypes including shoot length (SL), root length (RL), shoot fresh weight (SFW), root fresh weight (RFW), shoot dry weight (SDW), and root dry weight (RDW) under 50 and 100 mM NaCl stress. QTL analysis identified a total of 38 QTLs for these 6 traits under two distinct concentrations of salt, distributing on chromosomes 1, 2, 3, 4, 5, 6, 7 and 10. A total of 21 QTLs were found in 6 stable loci. A novel major QTL, qSL7 for shoot length on chromosome 7 was identified in two distinct concentrations. A chromosome segment substitution line (CSSL) harboring the qSL7 locus from PA64s with 93-11 background was developed and exhibited higher SL value, higher K + concentration, and lower Na + concentration compared to 93-11. With BC 5 F 2:3 derived from CSSL-qSL7/93-11, the qSL7 was fine mapped within a 252.9 kb region on chromosome 7 where 40 annotated genes located including, LOC_Os07g43530, which encodes a DNA-binding domain containing protein reported previously as a transcription factor playing a positive role in salt stress tolerance. Our study provides new genetic resources for improvement of salinity tolerance in rice breeding.
Synaptogenesis is considered necessary for learning and memory. Recently, it has been suggested that progesterone (PROG) effects synaptogenesis of the cerebellar Purkinje cell, helps alleviate symptoms of multiple sclerosis and helps arrest spinal cord neurodegeneration. However, it is unclear whether PROG influences synaptic plasticity in central nervous system neurons after global cerebral ischemia. The purpose of the present study was to reveal PROG's influence on postischemic synaptogenesis in the CA1 region of the hippocampus in rats. Global cerebral ischemia was induced in male Sprague-Dawley rats by the 4-vessel occlusion (4-VO) method. To determine the amount of synaptogenesis, growth-associated protein 43 (GAP-43) and synaptophysin (SYP) expression were examined by immunohistochemical, reverse transcription-polymerase chain reaction (RT-PCR) and western blot techniques. Histological and behavioral tests were used to indicate the effect of PROG on global cerebral ischemia. Recovery times were 3, 7, 14, 21, and 35 days after surgery. We found that PROG increased the expression of GAP-43 and SYP. In addition, there was a significant increase in neuronal cell density and improvement ability to remain on an accelerating rotarod observed in the 4-VO rats treated with PROG compared to vehicle. We propose that PROG helped enable synaptogenesis in the CA1 region of therat hippocampus after global cerebral ischemia in rats.
Genetic erosion refers to the loss of genetic variation in a crop. In China, only a few original landraces of rice (Oryza sativa) were used in breeding and these became the primary genetic background of modern varieties. Expanding the genetic diversity among Chinese rice varieties and cultivating high-yielding and high-quality varieties with resistance to different biotic and abiotic stresses is critical. Here, we used the clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated protein9(Cas9) genome editing system to edit Semi-Dwarf1 (SD1) in the elite landraces Kasalath and TeTePu (TTP), which contain many desired agronomic traits such as tolerance to low phosphorous and broad-spectrum resistance to several diseases and insects. Mutations of SD1 confer shorter plant height for better resistance to lodging. Field trials demonstrated that the yield of the new Kasalath and TTP mutant lines was better than that of the wild type under modern cultivation and that the lines maintained the same desirable agronomic characteristics as their wild-type progenitors. Our results showed that breeding using available landraces in combination with genomic data of different landraces and gene-editing techniques is an effective way to relieve genetic erosion in modern rice varieties.
Background: In rice (Oryza sativa) and other grains, weak growth (dwarfism, short panicle length, and low seedsetting rate) and early senescence lead to reduced yield. The molecular mechanisms behind these processes have been widely studied; however, the complex genetic regulatory networks controlling growth and senescence require further elucidation. Results: We isolated a mutant exhibiting weak growth throughout development and early senescence of leaf tips, and designated this mutant weakness and leaf senescence5 (wls5). Histological analysis showed that the poor growth of wls5 plants involved a reduction in cell length and number. Physiological analysis and transmission electron microscopy revealed that the wls5 cells had abnormal chloroplasts, and the mutants underwent chlorophyll degradation triggered by accumulation of reactive oxygen species. Consistent with this, RNA sequencing revealed changes in senescencerelated gene expression in wls5 plants. The wls5 mutants also exhibited significantly higher stomatal density and altered phytohormone contents compared with wild-type plants. Fine mapping delimited WLS5 to a 29-kb region on chromosome 5. DNA sequencing of wls5 identified a 3-bp deletion in the first exon of LOC_Os05g04900, resulting in a deletion of a lysine in the predicted protein. Knockout of LOC_Os05g04900 in Nipponbare plants caused leaf senescence, confirming this locus as the causal gene for WLS5. Conclusions: We identified a novel mutant (wls5) that affects plant development and leaf senescence in rice. LOC_Os05g04900, encoding a protein of unknown function, is the causal gene for wls5. Further molecular study of WLS5 will uncover the roles of this gene in plant growth and leaf senescence.
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