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Supramolecular polymer-based gel fracturing fluid with a double network applied in ultra-deep hydraulic fracturing

Petroleum Science 2024年第3期21卷 1875-1888页

作者： yong-ping huang yong Hu Chang-Long Liu Yi-Ning Wu Chen-Wei Zou Li-Yuan Zhang Ming-Wei Zhao Cai-Li Dai School of Petroleum Engineering China University of Petroleum(East China)Qingdao266580ShandongPR China Shandong Key Laboratory of Oilfield Chemistry China University of Petroleum(East China)Qingdao 266580ShandongPR China SINOPEC Petroleum Exploration and Production Research Institute Beijing102206PR China CNOOC China Co Ltd Tianjin BranchTianjin300459PR China

A gel based on polyacrylamide,exhibiting delayed crosslinking characteristics,emerges as the preferred solution for mitigating degradation under conditions of high temperature and extended shear in ultralong *** viscosity/viscoelasticity of the fracturing fluid was required to maintain excellent proppant suspension properties before *** into account both the cost and the potential damage to reservoirs,polymers with lower concentrations and molecular weights are generally *** this work,the supramolecular action was integrated into the polymer,resulting in significant increases in the viscosity and viscoelasticity of the synthesized supramolecular polymer *** double network gel,which is formed by the combination of the supramolecular polymer system and a small quantity of Zr-crosslinker,effectively resists temperature while minimizing permeability damage to the *** results indicate that the supramolecular polymer system with a molecular weight of(268—380)×10^(4)g/mol can achieve the same viscosity and viscoelasticity at 0.4 wt%due to the supramolecular interaction between polymers,compared to the 0.6 wt%traditional polymer(hydrolyzed polyacrylamide,molecular weight of 1078×10^(4)g/mol).The supramolecular polymer system possessed excellent proppant suspension properties with a 0.55 cm/min sedimentation rate at 0.4 wt%,whereas the0.6 wt%traditional polymer had a rate of 0.57 cm/*** comparison to the traditional gel with a Zrcrosslinker concentration of 0.6 wt%and an elastic modulus of 7.77 Pa,the double network gel with a higher elastic modulus(9.00 Pa)could be formed only at 0.1 wt%Zr-crosslinker,which greatly reduced the amount of residue of the fluid after *** viscosity of the double network gel was66 m Pa s after 2 h shearing,whereas the traditional gel only reached 27 m Pa s.

关键词： Ultra-deep reservoir Gel fracturing fluid Double network Supramolecular polymer system Proppant suspension property

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CRISPR/Cas9-mediated targeted gene mutagenesis in Spodoptera litura

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Insect Science 2016年第3期23卷 469-477页

作者： Hong-Lun Bi Jun XU An-Jiang Tan yong-ping huang College of Life Science Fujian Agriculture and Forestry University Fuzhou Key Laboratory of Insect Developmental and EvolutionaryBiology Institute of Plant Physiology and Ecology Shanghai Institutes for Biological Sciences Chinese Academy of Sciences ShanghaiChina

Custom-designed nuclease technologies such as the clustered regularly in- terspaced short palindromic repeat （CRISPR）-associated （Cas） system provide attractive genome editing tools for insect functional genetics. The targeted gene mutagenesis me- diated by the CR1SPR/Cas9 system has been achieved in several insect orders including Diptera, Lepidoptera and Coleoptera. However, little success has been reported in agricul- tural pests due to the lack of genomic information and embryonic microinjection techniques in these insect species. Here we report that the CRISPR/Cas9 system induced efficient gene mutagenesis in an important Lepidopteran pest Spodoptera litura. We targeted the S. litura Abdominal-A （Slabd-A） gene which is an important embryonic development gene and plays a significant role in determining the identities of the abdominal segments of in- sects. Direct injection of Cas9 messenger RNA and Slabd-A-specific single guide RNA （sgRNA） into S. litura embryos successfully induced the typical abd-A deficient pheno- type, which shows anomalous segmentation and ectopic pigmentation during the larval stage. A polymerase chain reaction-based analysis revealed that the Cas9/sgRNA complex effectively induced a targeted mutagenesis in S. litura. These results demonstrate that the CRISPR/Cas9 system is a powerful tool for genome manipulation in Lepidopteran pests such as S. litura.

关键词： abdominal-A gene CRISPR/Cas9 system genome editing microinjection Spodoptera litura

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CRISPR/Cas9-mediated ebony knockout results in puparium melanism in Spodoptera litura

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Insect Science 2019年第6期26卷 1011-1019页

作者： Hong-Lun Bi Jun Xu Lin He yong Zhang Kai Li yong-ping huang College of Life Sciences East China Normal University Shanghai China Key Laboratory of Insect Developmental and Evolutionary Biology Institute of Plant Physiology and Ecology Shanghai Institutes for Biological Sciences Chinese Academy of SciencesShanghai China Department of Biology University of Nevada Reno NV USA

Insect body pigmentation and coloration are critical to adaption to the environment. To explore the mechanisms that drive pigmentation, we used the clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated protein 9 (Cas9) genome editing system to target the ebony gene in the non-model insect Spodoptera litura. Ebony is crucial to melanin synthesis in insects. By directly injecting Cas9 messenger RNA and ebony-specific guide RNAs into S. litura embryos, we successfully induced a typical ebony-deficient phenotype of deep coloration of the puparium and induction of melanin formation during the pupal stage. Polymerase chain reaction-based genotype analysis demonstrated that various mutations had occurred at the sites targeted in ebony. Our study clearly demonstrates the function of ebony in the puparium coloration and also provides a potentially useful marker gene for functional studies in S. litura as well as other lepidopteran pests.

关键词： CR1SPR/Cas9 ebony marker gene Spodoptera litura

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Leap forward with insect genomics

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Insect Science 2016年第3期23卷 332-334页

作者： Bin Chen Xian-Hui Wang Zhijian Tu yong-ping huang Institute of Entomology and Molecular Biology Chongqing Normal University Chongqing China State Key Laboratory of IntegratedManagement of Pest Insects & Rodents Institute of Zoology Chinese Academy of Sciences Beijing China Department of Biochemistry Fralin Life Science Institute and Program in Genetics Bioinformatics and Computational Biology Virginia Tech Blacksburg Virginia USAand Key Laboratory of Insect Development and Evolutionary Biology Institute of Plant Physiology and Ecology Shanghai Institutes forBiological Sciences Chinese Academy of Sciences Shanghal China

Insects comprise the largest animal group in the world. More than a million insect species have been identified, leaving possibly two to three times more to be discovered. Systematic understanding of the genetic content of an insect species began more than 15 years ago when the genome sequence of the fruit fly, Drosophila melanogaster was reported （Adams et al., 2000）. For the first time, scientists were able to glean unprecedented insights into the genetic blueprint of a model insect, presenting its 13 600 protein coding genes. Such fundamental information opened a floodgate for comparative and functional analysis of these genes and enabled systems approaches to comparative, evolutionary and population genomic studies. In recent years, next-generation and third-generation sequencing platforms brought genome sequencing to more than a hundred insect species, reach- ing far beyond the few well-studied model organisms. The international consortium for insect genomes is work- ing toward an ambitious goal to sequence 5000 species of insects.

关键词： Drosophila melanogaster Insect genetics DNA sequencing Genetic code Insect evolution Insect population genetics

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Disruption of egg-specific protein causes female sterility in Bombyx mori

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Insect Science 2022年第1期29卷 128-138页

作者： Xia Xu Yao-Hui Wang Zu-Lian Liu yong-Qiang Wang Lin He Kai Li yong-ping huang Institute of Sericulture and Tea Research Zhejiang Academy of Agricultural SciencesHangzhou310021China Key Laboratory of Insect Developmental and Evolutionary Biology CAS Center for Excellence in Molecular Plant SciencesShanghai Institute of Plant Physiology and EcologyChinese Academy of SciencesShanghai200032China School of Life Science East China Normal UniversityShanghai200062China

Yolk proteins are the main source of nutrients during embryonic and early larval development in oviparous animals. Therefore, vitellogenesis is crucial for reproduction. The silkworm, Bombyx mori, is a model lepidopteran insect in which there are three yolk proteins: vitellin, 30-kDa protein, and egg-specific protein (Esp). In this study, we explored the gene function of Esp through transgenic clustered regularly interspaced palindromic repeats (CRISPR) / CRISPR-associated protein 9 technology-mediated mutations in the silkworm. We found that Esp mutation resulted in female sterility but had no effect on male fertility. Female mutants could lay eggs after mating, but the eggs were smaller and lighter colored than those laid by wild-type females. The most important finding is that the eggs laid by female mutants did not hatch. Furthermore, we observed stable inheritance of female sterility caused by Esp mutation through successive generations. Thus, Esp encodes a yolk protein that is crucial for female reproductive success and is a potential target for pest control.

关键词： Bombyx mori CRISPR/Cas9 Esp female sterility

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CRISPR disruption of TCTP gene impaired normal development in the silkworm Bombyx mori

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Insect Science 2019年第6期26卷 973-982页

作者： Zu-Lian Liu Jun Xu Lin Ling Ru Zhang Peng Shang yong-ping huang Faculty of Life Sciences Northwestern Polytechnical University Xi'anChina Key Laboratory of Insect Developmental and Evolutionary Biology Institute of Plant Physiology and Ecology Shanghai Institutes for Biological Sciences Chinese Academy of Sciences Shanghai China

The translationally controlled tumor protein (TCTP) is a highly conserved and multifunctional protein with activities ranging from cytoskeletal regulation to transcription regulation in numerous organisms. In insects, TCTP is essential for cell growth and proliferation. Recently, TCTP has been reported to affect the innate intestinal immune pathway in the Bombyx mori silkworm, a lepidopteran model insect. However, the comprehensive physiological roles of TCTP in the silkworm remain poorly understood. Here, we performed functional analysis of BmTCTP by using a binary transgenic CRISPR/Cas9 (clustered regularly interspaced short palindromic repeat/RNA-guided CRISPER-associated protein 9 nucleases) system. Disruption of BmTCTP led to developmental arrestment and subsequent lethality in third instar larvae. Histological analysis revealed that growth impairment originated from decreased cell size, and the proliferation and differentiation of intestinal epithelial cells were also affected. RNA-seq analysis revealed that genes involved in carbohydrate metabolism, lipid metabolism and digestive system pathways were significantly affected by BmTCTP depletion. Together, the results demonstrated that BmTCTP plays a key role in controlling larval growth and development.

关键词： Bombyx mori CRISPR/Cas9 TCTP

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BmHpo mutation induces smaller body size and late stage larval lethality in the silkworm,Bombyx mori

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Insect Science 2018年第6期25卷 1006-1016页

作者： Xia Xu Hong-Lun Bi Zhong-Jie Zhang Yang Yang Kai Li yong-ping huang yong Zhang Lin He School of Life Science East China Normal UniversityShanghaiChina Key Laboratory of Insect Developmental and Evolutionary Biology Institute of Plant Physiology and EcologyShanghai Institutes for Biological SciencesChinese Academy of SciencesShanghaiChina

As a core member of the Hippo signaling pathway,Hpo plays a critical role in regulating growth and *** studies reported that loss of function of Hpo results in increased proliferation,reduced apoptosis and induction of tissue overgrowth in *** this study,we used CRISPR/Cas9(Clustered Regularly Interspaced Short Palindromic Repeats/Casg)to study Hpo gene (BmHpo)function in the lepidopteran insect Bombyx mori,known commonly as the *** analysis of BmHpo revealed an array of deletions in *** found that BmHpo knockout resulted in defects in body size regnlation,in developmental defects and pigment accumulation and early death. Our data show that BmHpo is essential for regulation of insect growth and development and that CRISPR/Cas9technology can serve as a basis for functional analysis of target genes in lepidopteran insects.

关键词： Bombyx mori CRISPR/Cas9 developmental defects Hippo signaling path- way Hpo

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MicroRNA-2738 regulates gene expression in the sex determination pathway in Bombyx mori

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Insect Science 2020年第4期27卷 646-654页

作者： Zu-Lian Liu Jun Xu Lin Ling De-Hong Yang Shu-Qing Chen yong-ping huang CAS Key Laboratory of Insect Developmental and Evolutionary Biology CAS Center for Excellence in Molecular Plant SciencesInstitute of Plant Physiology and EcologyCASShanghaiChina

MicroRNAs(miRNAs)are a class of short,non-coding transcripts that bind to B'-untranslated regions to trigger messenger RNA degradation or translational *** we explored how miRNAs regulate sex determination in Bombyx mori,a lepidopteran model *** known to be involved in sex determination,BmPSI,Bmdsx,and BmMasc\are predicted targets of the species-specific *** a dual luciferase reporter assay in HEK293T cells,we confirmed that miR-2738 suppressed transcription of BmPSl,Bmdsx,and *** levels of BniPSI and BmMasc were significantly down-regulated in *** miR-2738 *** contrast,the genetic disruption of miR-2738 using the clustered regularly interspaced short palindromic repeats(CRISPR)/CRISPR-associated protein 9 transgenic system increased the levels of BmPSI and BmMasc transcripts,whereas splicing of Bmdsx was unaltered by miR-2738 depletion or *** together,this study implicates miR-2738 as a minor regulator of sex determination genes in the silkworm.

关键词： CRISPR/Cas9 miRNA sex determination silkworm

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Intracolonial differences in gut bacterial community between worker and soldier castes of Coptotermes formosanus

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Insect Science 2012年第1期19卷 86-95页

作者： Hui Xiang Lei Xie Jun Zhang Yan-Hua Long Ning Liu yong-ping huang Qian Wang Institute of Plant Physiology and Ecology Shanghai Institutes for Biological SciencesChinese Academy of SciencesShanghaiChina

The establishment of symbiotic relationships with intestinal microorganisms enables termites to thrive on recalcitrant substrates such as cellulose and wood. A termite colony is composed of several different castes which have distinct feeding habits. The soldiers, for example, cannot feed by themselves and depend on workers, who feed them with digested or semi-digested foods. To investigate the influence of feeding habits on the bacterial symbionts, a comparative study of gut bacteria between worker and soldier castes of the termite Coptotermesformosanus was conducted. The bacterial communities of both castes were investigated using denaturing gradient gel electrophoresis （DGGE） and clonal analysis of 16S ribosomal DNA （rDNA）. Both methods indicated Bacteroidetes was the common predominant group; the common dominant phylotype was affiliated with a reported uncultured Bacteroidetes phylotype （BCfl-03）. There were significant differ- ences in Bacteroidetes and Spirochaetes between two castes. Compared to the gut bacteria of workers, those of soldiers were lower in abundance and diversity of Bacteroidetes and slightly higher in Spirochaetes. Two phylotypes （W8, W11） affiliated to Bacteroidetes and two （W26, W29） affiliated to Spirochaetes were exclusively found in the DGGE profile of the worker caste. Bacteroidetes are assumed to be involved in fermentation of sugars and nitrogenous compounds as well as degradation of uric acid. Spirochaetes are supposed to aid in the functions of acetogenesis and N2-fixation. The different feeding habits between workers and soldiers of C. formosanus may explain the observed differences in the gut bacterial community.

关键词： Coptotermesformosanus gut bacteria community 16S rDNA

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Dynamics of the intestinal bacterial community in black soldier fly larval guts and its influence on insect growth and development

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Insect Science 2023年第4期30卷 947-963页

作者： Xin-Yu Li Cheng Mei Xing-Yu Luo Dilinuer Wulamu Shuai Zhan yong-ping huang Hong Yang Hubei Key Laboratory of Genetic Regulation and Integrative Biology Institute of EntomologySchool of Life SciencesCentral China Normal UniversityWuhanChina CAS Key Laboratory of Insect Developmental and Evolutionary Biology CAS Center for Excellence in Molecular Plant SciencesInstitute of Plant Physiology and EcologyShanghai Institutes for Biological SciencesChinese Academy of SciencesShanghaiChina

Black soldier fly(BSF),Hermetia illucens(Diptera:Stratiomyidae),is a promi-nent insect for the bioconversion of various organic *** a saprotrophic insect,the BSF inhabits microbe-rich ***,the influences of the intestinal mi-croorganisms on BSF growth and development are not very *** this study,the dy-namics of the intestinal bacterial community of BSF larvae(BSFL)were analyzed using ***,Bacteroidetes,Firmicutes,and Proteobacteria were the most prevalent bacterial phyla in the intestines of all larval *** dynamic changes in bacterial community compositions among different larval instars were striking at the genus ***,Clostridium,Providencia,and Dysgonomonas were the relatively most abundant bacteria in the 1st-to 4th-instar BSFL,*** and Providencia also dominated the 5th-and 6th-instar larvae,at ratios of 31.1%and 47.2%,*** total,148 bacterial strains affiliated with 20 genera were isolated on differ-ent media under aerobic and anaerobic *** them,6 bacteria,BSF1-BSF6,were selected for further *** inoculation of the 6 isolates independently into germ-free BSFL feeding on an artificial diet showed that all the bacteria,except BSF4,sig-nificantly promoted BSF growth and development compared with the germ-free ***,Dysgonomonas,Klebsiella,Ochrobactrum,and Providencia promoted BSF development significantly by increasing the weight gains of larvae and pupae,as well as increasing the prepupae and eclosion *** addition,Citrobacter,Klebsiella and Prov-idencia shortened the BSF life cycle *** results illustrate the promotive effects of intestinal bacteria on BSF growth and development.

关键词： bacterial isolates germ-free BSF larvae growth and development Hermetia illucens promotive effects

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