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Significant changes in arbuscular mycorrhizal community and soil physicochemical properties during the saline-alkali grassland vegetation succession

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BIOCELL 2022年第11期46卷 2475-2488页

作者： yajie liu LINLIN FANG CHUNXUE YANG College of Landscape Architecture Northeast Forestry UniversityHarbin150040China

Arbuscular mycorrhizal(AM)fungi are widely distributed in various habitats,and the community composition varies in response to the changing environmental conditions.To explore the response of community composition to the succession of saline-alkali land,soil samples were collected from three succession stages of Songnen saline-alkali grassland.Subsequently,the soil characteristics were determined and the AM fungi in soil samples were analyzed by high-throughput sequencing.Then,the response relationship between community composition and soil characteristics was studied by Canonical correlation and Pearson analyses.The soil properties improved with the succession of saline-alkali grassland.There was no significant difference in alpha diversity between the first and second succession stage(Suaeda glauca and Puccinellia tenuiflora,respectively),and the microbial community had a dense association network at the third stage(Leymus chinensis);in addition,each succession stage had significantly enriched amplicon sequence variants(ASVs)and functional pathways.All the soil properties except cellulase activity had significant effects on community composition.Furthermore,the pH,organic carbon,organic matter,and sucrase activity significantly correlated with alpha diversity indices.These results provide a theoretical basis for realizing the significant changes in AM fungal community and soil properties during the saline-alkali grassland vegetation succession.

关键词： AM fungal community High-throughput sequencing Succession of saline-alkali land Soil physicochemical characteristics

A single-domain protein catenane of dihydrofolate reductase

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National Science Review 2023年第11期10卷 128-137页

作者： Jing Fang Tianzuo Li Jiyeon Lee Dahye Im Lianjie Xu yajie liu Jongcheol Seo Wen-Bin Zhang Beijing National Laboratory for Molecular Sciences Key Laboratory of Polymer Chemistry & Physics of Ministry of EducationCenter for Soft Matter Science and EngineeringCollege of Chemistry and Molecular EngineeringPeking University Department of Chemistry Pohang University of Science and Technology (POSTECH) Beijing Academy of Artificial Intelligence

A single-domain protein catenane refers to two mechanically interlocked polypeptide rings that fold synergistically into a compact and integrated structure, which is extremely rare in nature. Here, we report a single-domain protein catenane of dihydrofolate reductase(cat-DHFR). This design was achieved by rewiring the connectivity between secondary motifs to introduce artificial entanglement and synthesis was readily accomplished through a series of programmed and streamlined post-translational processing events in cells without any additional in vitro reactions. The target molecule contained few exogenous motifs and was thoroughly characterized using a combination of ultra-performance liquid chromatography–mass spectrometry, sodium dodecyl sulfate–polyacrylamide gel electrophoresis, protease cleavage experiments and ion mobility spectrometry–mass spectrometry. Compared with the linear control, cat-DHFR retained its catalytic capability and exhibited enhanced stability against thermal or chemical denaturation due to conformational restriction. These results suggest that linear proteins may be converted into their concatenated single-domain counterparts with almost identical chemical compositions, well-preserved functions and elevated stabilities, representing an entirely new horizon in protein science.

关键词： cellular synthesis catenane protein domain chemical topology DHFR

Arbuscular Mycorrhizal Fungal Colonization at Different Succession Stages in Songnen Saline-Alkali Grassland

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Phyton-International Journal of Experimental Botany 2023年第1期92卷 297-310页

作者： yajie liu Yunhui Zhou Linlin Fang Chunxue Yang College of Landscape Architecture Northeast Forestry UniversityHarbin150040China

Arbuscular mycorrhizal(AM)fungi can form symbiosis with 90%of the vascular plants and play important roles in ecosystem.To realize the AM fungal colonization at different succession stages in saline-alkali land and screen AM fungi species with great functions,roots and soil samples were collected from the three succession stages of Songnen saline-alkali grassland.The soil properties and AM fungal colonization were measured,and the fungus distributed extensively in three stages was annotated by sequencing for AML1/AML2 target,subsequently,maize was selected as the host to verify its colonization.The results showed that the soil properties improved with the succession of saline-alkali grassland.The plants’communities of the three stages could be colonized by AM fungi,and the colonization rate of Leymus chinensis(the third stage)ranged from 66.67%to 100%,Puccinellia tenuiflora(the second stage)ranged from 50%to 80%,while the Suaeda glauca(the first stage)was only 35%–60%.Glomeraceae sp1 was identified as the dominant AM fungi species which occurred frequently in the succession of saline-alkali land with the isolation frequency,relative abundance,and importance value of 100%,18.1%,and 59.1%,respectively.The colonization rate of Glomeraceae sp1 in maize ranged from 80%to 87%and similar mycorrhizal characteristics were detected in the roots of P.tenuiflora,S.glauca,and L.chinensis,indicating that Glomeraceae sp1 colonized the samples in the field.The correlation matrix indicated that colonization rate,colonization intensity,and vesicle abundance were closely related to soil conditions most,and they were related significantly to all the soil properties except cellulase activity.Besides,redundancy analysis(RDA)showed that soil properties drove the changes of AM fungal colonization and sporulation.These results will provide theoretical support for realizing the relationship between AM fungal colonization and soil conditions,and also for the exploration of AM fungi species with great functions.

关键词： Succession of saline-alkali land soil properties AM fungi colonization characteristics

Effects of Different Arbuscular Mycorrhizal Fungi on Physiology of Viola prionantha under Salt Stress

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Phyton-International Journal of Experimental Botany 2023年第1期92卷 55-69页

作者： yajie liu Linlin Fang Wenna Zhao Chunxue Yang College of Landscape Architecture Northeast Forestry UniversityHarbin150040China

Arbuscular mycorrhizal(AM)fungi distribute widely in natural habits and play a variety of ecological functions.In order to test the physiological response to salt stress mediated by different AM fungi,Viola prionantha was selected as the host,the dominant AM fungus in the rhizosphere of V.philippica growing in Songnen saline-alkali grassland,Rhizophagus irregularis,and their mixtures were used as inoculants,and NaCl stress was applied after the roots were colonized.The results showed that V.philippica could be colonized by AM fungi in the field and the colonization rate ranged from 73.33%to 96.67%,and Claroideoglomus etunicatum was identified as the dominant AM fungi species in the rhizosphere of V.philippica by morphology combined with sequencing for AM fungal AML1/AML2 target.Inoculation with both the species resulted in the formation of mycorrhizal symbiosis(the colonization rate was more than 70%)and AM fungi significantly enhanced plants’tolerance to salt stress of varying magnitude.Higher activity of antioxidant enzymes and augmented levels of proline and other osmoregulators were observed in AM plants.The content of MDA in CK was higher than that in the inoculations with the stress of 100,200,and 250 mM.All indices except soluble protein content and MDA content were significantly correlated with AM fungal colonization indices.The analysis for different AM fungal effects showed that the mixtures and R.irregularis worked even better than C.etunicatum.These results will provide theoretical support for the exploration and screening of salt-tolerant AM fungi species and also for the application of AM-ornamental plants in saline-alkali urban greening.

关键词： Dominant AM fungi species Viola philippica salt stress Viola prionantha physiological response

Claroideoglomus etunicatum improved the growth and saline-alkaline tolerance of Potentilla anserina by altering physiological and biochemical properties

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BIOCELL 2022年第8期46卷 1967-1978页

作者： YUNHUI ZHOU yajie liu YUDAN WANG CHUNXUE YANG College of Landscape Architecture Northeast Forestry UniversityHarbin150040China

To investigate the effects of arbuscular mycorrhizal(AM)fungi on the growth and saline–alkaline tolerance of Potentilla anserina L.,the seedlings were inoculated with Claroideoglomus etunicatum(W.N.Becker&Gerd.)C.Walker&A.Schüßler in pot cultivation.After 90 days of culture,saline–alkaline stress was induced with NaCl and NaHCO_(3)solution according to the main salt components in saline–alkaline soils.Based on the physiological response of P.anserina to the stress in the preliminary experiment,the solution concentrations of 0 mmol/L,75 mmol/L,150 mmol/L,225 mmol/L and 300 mmol/L were treated with stress for 10 days,respectively.The mycorrhizal colonization rate,mycorrhizal dependence,chlorophyll content,malondialdehyde content,antioxidant enzyme activities,osmoregulation substances content and water status were measured.The results showed that with the increase of NaCl and NaHCO_(3)stress concentration,mycorrhizal colonization rate,colonization intensity,arbuscular abundance and vesicle abundance decreased,and reached the lowest value at 300 mmol/L.Strong mycorrhizal dependence was observed after the symbiosis with AM fungus,and the dependence was higher under NaHCO_(3)treatment.Under NaCl and NaHCO_(3)stress,inoculation with AM fungus could increase chlorophyll content,decrease malondialdehyde content,increase activities of superoxide dismutase,peroxidase and catalase,increase contents of proline,soluble sugar and soluble protein,increase tissue relative water content and decrease water saturation deficit.It was concluded that salt–alkali stress inhibited the colonization of AM fungus,but the mycorrhiza still played a positive role in maintaining the normal growth of plants under salt–alkali stress.

关键词： Arbuscular mycorrhizal fungi Colonization characteristics Antioxidant system Osmoregulation substances Water status

Rational Design and Cellular Synthesis of Proteins with Unconventional Chemical Topology

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Chinese Journal of Chemistry 2023年第21期41卷 2873-2880页

作者： Tianzuo Li Fan Zhang Jing Fang yajie liu Wen-Bin Zhang Beijing National Laboratory for Molecular Sciences Key Laboratory of Polymer Chemistry&Physics of Ministry of EducationCenter for Soft Matter Science and EngineeringCollege of Chemistry and Molecular EngineeringPeking UniversityBeijing100871 China Beijing Academy of Artificial Intelligence Beijing100084 China

Chemical topology refers to the three-dimensional arrangement(i.e.,connectivity and spatial relationship)of a molecule's constituent atoms and bonds.The molecular mechanism for translation defines the linear configuration of all nascent proteins.Nontrivial protein topology arises only upon post-translational processing events and often imparts functional benefits such as enhanced stability,making topology a unique dimension for protein engineering.Utilizing the assembly-reaction synergy,our group has developed several methods for the effective and convenient cellular synthesis of a variety of topological proteins,such as lasso proteins,protein rotaxanes,and protein catenanes.The work opens the access to new protein classes and paves the road toward illustrating the topological effects on structure-function relationship of proteins,which lays solid foundation for exploring topological proteins’practical application.

关键词： Active template Catenane Cellular synthesis Chemical topology Lasso protein Protein engineering

Isolation and species diversity of arbuscular mycorrhizal fungi in the rhizosphere of Puccinellia tenuiflora of Songnen saline-alkaline grassland,China

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BIOCELL 2022年第11期46卷 2465-2474页

作者： YUNHUI ZHOU yajie liu WENNA ZHAO FEI CHEN YUDAN WANG CHUNXUE YANG College of Landscape Architecture Northeast Forestry UniversityHarbin150040China

Salinization has led to the deterioration of the ecological environment,affected the growth of plants,and hindered the development of agriculture and forestry.Arbuscular mycorrhizal(AM)fungi,as important soil microorganisms,play significant physiological and ecological roles in promoting plant nutrient absorption and improving soil structure.Puccinellia tenuiflora(Turcz.)Scribn.et Merr.in Songnen saline-alkaline grassland was selected as the research object to observe AM fungal colonization of the roots and explore the species and diversity of AM fungi in symbiotic association with P.tenuiflora.This study showed that AM fungi colonized in P.tenuiflora roots and formed a typical Arum-type mycorrhizal structure.A significant correlation was observed between vesicular abundance and the colonization intensity of mycorrhiza.Isolation and identification revealed 40 species of AM fungi in the rhizosphere of P.tenuiflora,belonging to 14 genera,of which two species could not be identified.The richness of the genus Glomus was the highest,accounting for 30%of the total species.Funneliformis mosseae and Rhizophagus intraradices were isolated from all the samples and were the species with the widest distribution in the rhizosphere of P.tenuiflora.Correlation analysis showed that pH only had a significant impact on the distribution of a few species,such as Glomus pustulatum,Diversispora spurca,Glomus aggregatum,Rhizophagus clarum,and Acaulospora foveata.The present study provides a theoretical basis for further exploring the resources of AM fungi in saline-alkaline soil.

关键词： Morphological identification Spore density Species richness Diversity indexes Colonization intensity

A Moonlighting Superpositively Charged SpyCatcher

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CCS Chemistry 2023年第11期5卷 2663-2673页

作者： Tingting Yang yajie liu Wen-Hao Wu Rongmei Peng Wen-Bin Zhang Beijing National Laboratory for Molecular Sciences Key Laboratory of Polymer Chemistry and Physics of Ministry of EducationCenter for Soft Matter Science and EngineeringCollege of Chemistry and Molecular EngineeringPeking UniversityBeijing 100871 Key Laboratory of Vision Loss and Restoration Ministry of EducationBeijing 100084 Department of Ophthalmology Peking University Third HospitalBeijing 100191 Beijing Academy of Artificial Intelligence Beijing 100084

Genetically encoded covalent peptide tagging tools such as the SpyTag/SpyCatcher reactive pair,have been demonstrated versatile and useful for protein modification.Herein,we present a superpositively charged SpyCatcher bearing a theoretical net charge of+21 capable of accomplishing multiple unrelated independent tasks to enrich this toolbox and cultivate new functions.The SpyCatcher(+21)possessed stimuli-responsive reactivity toward SpyTag and could serve as a potent and general platform for the delivery of proteins,including RNaseAinto HeLa cells.Remarkably,the delivered RNase A caused substantial proliferation inhibition toward HeLa cells.In addition,the superpositively charged SpyCatcher could form coacervate with plasmid DNA for further study of gene delivery and liquid–liquid phase separation.These findings demonstrate the robustness of the SpyTag/SpyCatcher structure against surface mutation and the prospect of applying supercharging technology on diverse functional proteins to create moonlighting proteins.

关键词： SpyCatcher stimuli-responsive protein delivery phase separation supercharging cyclic proteins topology

一氧化氮参与调控油菜素内酯增强高山离子芥悬浮细胞抗寒性

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植物研究 2024年第1期44卷 118-131页

作者：刘亚洁安黎哲兰州大学草种创新与草地农业生态系统全国重点实验室/农业农村部草牧业创新重点实验室/草地农业教育部工程研究中心/草地农业科技学院兰州730020 兰州大学生命科学学院兰州730000 北京林业大学生态与自然保护学院北京100083

为探究油菜素内酯(BRs)诱导提高植物的抗寒性是否受一氧化氮(NO)信号分子调控,以高山离子芥(Chorispora bungeana)悬浮细胞为材料,分别用24-表油菜素内酯(EBR)、NO供体SNP、NO清除剂PTIO、一氧化氮合成酶(NOS)抑制剂L-NAME、EBR+PTIO以... 详细信息

为探究油菜素内酯(BRs)诱导提高植物的抗寒性是否受一氧化氮(NO)信号分子调控,以高山离子芥(Chorispora bungeana)悬浮细胞为材料,分别用24-表油菜素内酯(EBR)、NO供体SNP、NO清除剂PTIO、一氧化氮合成酶(NOS)抑制剂L-NAME、EBR+PTIO以及EBR+L-NAME进行处理,分析在低温胁迫下以上处理对细胞抗寒性、活性氧(ROS)水平以及抗氧化防御系统的影响。结果表明:(1)外源EBR处理可以缓解低温胁迫对悬浮细胞活力的抑制以及离子渗漏和膜脂过氧化程度的加剧,从而增强细胞对低温的抗逆能力。SNP处理对上述生理指标的影响与EBR类似。(2)在低温胁迫下,与EBR处理细胞相比,EBR+PTIO以及EBR+L-NAME处理会导致悬浮细胞活力下降,离子渗漏和膜脂过氧化程度显著升高,表明阻断NO信号会降低EBR诱导提高的抗寒性。(3)与仅受低温胁迫的细胞相比,外源EBR处理导致细胞NO含量和NOS活性进一步升高,而EBR诱导的NO积累可以被PTIO或L-NAME所抑制。(4)EBR、SNP处理均可以明显抑制低温胁迫下离子芥悬浮细胞过氧化氢(H_(2)O_(2))含量、超氧阴离子(O_(2)·^(-))产生速率和羟自由基(OH-)含量的升高,并显著增强抗坏血酸过氧化物酶(APX)、过氧化氢酶(CAT)、谷胱甘肽还原酶(GR)、过氧化物酶(POD)和超氧化物歧化酶(SOD)的活性以及增加抗坏血酸(AsA)和还原性谷胱甘肽(GSH)的含量,从而缓解低温造成的氧化损伤,而PTIO、L-NAME会不同程度抑制低温胁迫下EBR的这些保护功能。综上所述,EBR通过激活离子芥悬浮细胞的NOS活性来促进细胞内源NO积累,从而提高其抗寒性。EBR处理可以抑制低温胁迫下ROS过量积累并增强细胞的抗氧化防御能力,这两个过程都受NO信号分子调控,从而缓解低温造成的氧化损伤,防止膜脂过氧化程度加剧,提高细胞对低温胁迫的抗逆性。因此,低温胁迫下,高山离子芥悬浮细胞NOS来源的NO是EBR信号转导的下游信号分子。

关键词：油菜素内酯一氧化氮高山离子芥抗寒性活性氧抗氧化防御系统