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Switchgrass (Panicum virgatum L.) is a promising perennial bioenergy crop that achieves high yields with relatively low nutrient and energy inputs. Modification of cell wall composition for reduced recalcitrance can lower the costs of deconstructing biomass to fermentable sugars and other intermediates. We have engineered overexpression of OsAT10, encoding a rice BAHD acyltransferase and QsuB, encoding dehydroshikimate dehydratase from Corynebacterium glutamicum, to enhance saccharification efficiency in switchgrass. These engineering strategies demonstrated low lignin content, low ferulic acid esters, and increased saccharification yield during greenhouse studies in switchgrass and other plant species. In this work, transgenic switchgrass plants overexpressing either OsAT10 or QsuB were tested in the field in Davis, California, USA for three growing seasons. No significant differences in the content of lignin and cell wall-bound p-coumaric acid or ferulic acid were detected in transgenic OsAT10 lines compared with the untransformed Alamo control variety. However, the transgenic overexpressing QsuB lines had increased biomass yield and slightly increased biomass saccharification properties compared to the control plants. This work demonstrates good performance of engineered plants in the field, and also shows that the cell wall changes in the greenhouse were not replicated in the field, emphasizing the need to validate engineered plants under relevant field conditions.

The widely used rice variety Lijiangxintuanheigu (LTH) shows a universal susceptibility to thousands of Magnaporthe oryzae isolates, the causal agent of devastating rice blast, making LTH an ideal line in resistance (R) gene cloning. However, the underlying genetic mechanism of the universal susceptibility has not been fully revealed because of the lack of a high-quality genome. Here, we took a genomic approach together with experimental assays to investigate LTHs universal susceptibility to rice blast. Using Nanopore long reads, we assembled a chromosome-level genome. Millions of genomic variants were detected by comparing LTH with 10 other rice varieties, of which large-effect variants could affect plant immunity. Gene family analyses show that the number of R genes and leucine-rich repeat receptor-like protein kinase (LRR-RLK)-encoding genes decrease significantly in LTH. Rice blast resistance genes called Pi genes are either absent or disrupted by genomic variations. Additionally, residual R genes of LTH are likely under weak pathogen selection pressure, and other plant defense-related genes are weakly induced by rice blast. In contrast, the pattern-triggered immunity (PTI) of LTH is normal, as demonstrated by experimental assays. Therefore, we conclude that weak effector-trigger immunity (ETI)-mediated primarily by Pi genes but not PTI results in the universal susceptibility of LTH to rice blast. The attenuated ETI of LTH may be also associated with reduced numbers of R genes and LRR-RLKs, and minimally functional residual defense-related genes. Finally, we demonstrate the use of the LTH genome by rapid cloning of the Pi gene Piak from a resistant variety.

BackgroundBreeding for genes controlling key agronomic traits is an important goal of rice genetic improvement. To gain insight into genes controlling grain morphology, we screened M3 plants derived from 1,000 whole-genome sequenced (WGS) M2 Kitaake mutants to identify lines with altered grain size.ResultsIn this study, we isolated a mutant, named fast-neutron (FN) 60-4, which exhibits a significant reduction in grain size. We crossed FN60-4 with the parental line Kitaake and analyzed the resulting backcross population. Segregation analysis of 113 lines from the BC2F2 population revealed that the mutant phenotype is controlled by a single semi-dominant locus. Mutant FN60-4 is reduced 20% in plant height and 8.8% in 1000-grain weight compared with Kitaake. FN60-4 also exhibits an 8% reduction in cell number and a 9% reduction in cell length along the vertical axis of the glume. We carried out whole-genome sequencing of DNA pools extracted from segregants with long grains or short grains, and revealed that one gene, LOC_Os09g02650, cosegregated with the grain size phenotype in the BC1F2 and BC2F2 populations. This mutant allele was named grain shape 9-1 (gs9-1). gs9-1 carries a 3-bp deletion that affects two amino acids. This locus is a new allele of the BC12/GDD1/MTD1 gene that encodes a kinesin-like protein involved in cell-cycle progression, cellulose microfibril deposition and gibberellic acid (GA) biosynthesis. The GA biosynthesis-related gene KO2 is down-regulated in gs9-1. The dwarf phenotype of gs9-1 can be rescued by adding exogenous GA3. In contrast to the phenotypes for the other alleles, the gs9-1 is less severe, consistent with the nature of the mutation, which does not disrupt the open reading frame as observed for the other alleles.ConclusionsIn this study, we isolated a mutant, which exhibits altered grain shape and identified the mutated gene, gs9-1. Our study reveals that gs9-1 is a semi-dominant gene that carries a two-amino acid mutation. gs9-1 is allelic to the BC12/GDD1/MTD1 gene involved in GA biosynthesis. These results demonstrate the efficiency and convenience of cloning genes from the whole-genome sequenced Kitaake mutant population to advance investigations into genes controlling key agronomic traits in rice.

BackgroundThe availability of thousands of complete rice genome sequences from diverse varieties and accessions has laid the foundation for in-depth exploration of the rice genome. One drawback to these collections is that most of these rice varieties have long life cycles, and/or low transformation efficiencies, which limits their usefulness as model organisms for functional genomics studies. In contrast, the rice variety Kitaake has a rapid life cycle (9 weeks seed to seed) and is easy to transform and propagate. For these reasons, Kitaake has emerged as a model for studies of diverse monocotyledonous species.ResultsHere, we report the de novo genome sequencing and analysis of Oryza sativa ssp. japonica variety KitaakeX, a Kitaake plant carrying the rice XA21 immune receptor. Our KitaakeX sequence assembly contains 377.6 Mb, consisting of 33 scaffolds (476 contigs) with a contig N50 of 1.4 Mb. Complementing the assembly are detailed gene annotations of 35,594 protein coding genes. We identified 331,335 genomic variations between KitaakeX and Nipponbare (ssp. japonica), and 2,785,991 variations between KitaakeX and Zhenshan97 (ssp. indica). We also compared Kitaake resequencing reads to the KitaakeX assembly and identified 219 small variations. The high-quality genome of the model rice plant KitaakeX will accelerate rice functional genomics.ConclusionsThe high quality, de novo assembly of the KitaakeX genome will serve as a useful reference genome for rice and will accelerate functional genomics studies of rice and other species.

BackgroundSwitchgrass (Panicum virgatum L.) is a promising bioenergy feedstock because it can be grown on marginal land and produces abundant biomass. Recalcitrance of the lignocellulosic components of the switchgrass cell wall to enzymatic degradation into simple sugars impedes efficient biofuel production. We previously demonstrated that overexpression of OsAT10, a BAHD acyltransferase gene, enhances saccharification efficiency in rice.ResultsHere we show that overexpression of the rice OsAT10 gene in switchgrass decreased the levels of cell wall-bound ferulic acid (FA) in green leaf tissues and to a lesser extent in senesced tissues, and significantly increased levels of cell wall-bound p-coumaric acid (p-CA) in green leaves but decreased its level in senesced tissues of the T0 plants under greenhouse conditions. The engineered switchgrass lines exhibit an approximate 40% increase in saccharification efficiency in green tissues and a 30% increase in senesced tissues.ConclusionOur study demonstrates that overexpression of OsAT10, a rice BAHD acyltransferase gene, enhances saccharification of lignocellulosic biomass in switchgrass.

The availability of a whole-genome sequenced mutant population and the cataloging of mutations of each line at a single-nucleotide resolution facilitate functional genomic analysis. To this end, we generated and sequenced a fast-neutron-induced mutant population in the model rice cultivar Kitaake (Oryza sativa ssp japonica), which completes its life cycle in 9 weeks. We sequenced 1504 mutant lines at 45-fold coverage and identified 91,513 mutations affecting 32,307 genes, i.e., 58% of all rice genes. We detected an average of 61 mutations per line. Mutation types include single-base substitutions, deletions, insertions, inversions, translocations, and tandem duplications. We observed a high proportion of loss-of-function mutations. We identified an inversion affecting a single gene as the causative mutation for the short-grain phenotype in one mutant line. This result reveals the usefulness of the resource for efficient, cost-effective identification of genes conferring specific phenotypes. To facilitate public access to this genetic resource, we established an open access database called KitBase that provides access to sequence data and seed stocks. This population complements other available mutant collections and gene-editing technologies. This work demonstrates how inexpensive next-generation sequencing can be applied to generate a high-density catalog of mutations.

Based on the case of the oil-gas separator for engine parts, this article has found the key steps to improve the cleanliness level by extracting and analyzing the impurity particles on its surface, and has reached the limit requirements in the enterprise standard. The whole detection process is divided into two parts: particle extraction and analysis. The impurity particles on the surface of the sample are extracted and collected by pressure washing, and the weight and size of the impurity particles are analyzed using a high-precision balance and a microscope. The test results show that the weight of the impurity particles on the surface of the oil and gas separator is 1.68 mg, and the maximum particle size is 887.31μm, which does not meet the product standard requirements. Through the analysis of the actual situation of the case to find the reasons and formulate measures, the final weight of the impurity particles on the surface of the improved sample is 1.22mg, and the maximum particle size is 481.40μm, which effectively improves the cleanliness level of the oil and gas separator.

At present, China has become the largest automobile production and sales country in the world, and the second largest lubricant consumption country. It is of great significance to establish China’s independent certification and evaluation system for lubricants.In recent years, China’s heavy-duty diesel engine has basically realized the technology autonomy, and the mode of lubricant evaluation standard equivalent to API specification is not suitable for China’s national conditions. It is imperative to develop a diesel engine oil quality evaluation research that is suitable for China’s national conditions. This paper mainly introduces a series of research on the performance difference of JAC 2.0TCI engine based on the physicochemical, wear and sediment evaluation of different reference oils under the condition of fuel dilution.

This paper selects 6 gasoline detergent synergists and 6 diesel detergent synergists that are mainstream in the market. The changes in mass, volume, hardness, size and surface morphology of the main components of the engine fuel supply system after being immersed in fuel containing detergent synergists for 28 days were studied,and these changes are compared with the benchmark fuel. The results show that most of the fuel detergent synergists have almost no corrosive effect on the engine fuel supply system components or are close to the reference fuel, but some gasoline detergent synergists have a slight corrosive swelling effect on the gasoline fuel supply system.

In this paper, a specific engine is selected to perform a bench test to evaluate the performance of the diesel lubricating oil. The injection time of the original engine is changed , and an appropriate after-injection strategy is selected to achieve accelerated aging of the lubricating oil.In the engine bench test, oil samples are taken at regular intervals to analyze the physical and chemical indicators and tribological characteristics, to determine the regularity of the change of the oil aging degree. At the same time, two different kinds of diesel lubricating oil were used for the test, so as to compare the performance parameters of different kinds of the lubricating oil during the aging process.

With the development of the society, the emission laws are increasingly strict, and EGR and multiple pre-injection strategy can effectively reduce emissions. In order to study the influence of EGR rate and pre-injection on the emission and performance of engine, a computational fluid dynamics (CFD) simulation model was established based on GT power. By changing the EGR rate, the number of pre-injection and the amount of pre-injection, the best technology coupling point is explored. The results show that EGR can effectively reduce the generation of NOx, but it will lead to the increase of soot production and the decrease of power; pre-injection can improve the trade-off relationship between NOx and power.

Based on different fuel injection strategies, this paper analyzes the factors such as engine original emission smoke, exhaust temperature, soot content, wear spot diameter and kinematic viscosity. The study found that delaying injection timing, increased afterburn, engine original soot emissions, exhaust gas temperature increase, but will increase the thermal load of the parts. At the same time, the growth rate of lubricant soot and kinematic viscosity increased; The wear spot diameter at the same soot content is reduced, and the wear is reduced. In the end, the paper finally selects 1°CA BTDC as the optimal fuel injection strategy to achieve rapid aging of engine lubricating oil in order to complete the assessment of the anti-wear performance of lubricating oil.

Based on two different components of lubricating oil, this paper analyzes the filtration efficiency, ash accumulation speed, vehicle WLTC emissions, fuel consumption, and CT scan on a National V emission standard engine equipped with a cGPF aftertreatment device. The results show that both oils can ensure that cGPF meets the emission and fuel consumption requirements of National VI emission standard, but oil F accumulates ash quickly before the ash accumulation of 33% in the discharge, which has a significant effect on PN reduction, and oil C accumulates ash faster after the accumulation of 33%. The effect of reducing the PN is accelerated. Oil F is based on a full calcium detergent system, which has a trend of first to slower for the collection efficiency; while oil C is based on a mixed Ca/Mg detergent system, which has a trend of first slowing and then fasting the collection efficiency. The critical point is generally about 33% of accumulated ash. For oil F, less ash is deposited in the front and middle of the cGPF, while more ash tends to deposit towards the outlet. In contrast, cGPF aged with Oil C has a relatively uniform deposition distribution.