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Identification of a key G-protein coupled receptor in mediating appressorium formation and fungal virulence against insects

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Science China(Life Sciences) 2021年第3期64卷 466-477页

作者： Junmei shang yanfang shang Guirong Tang Chengshu Wang Key Laboratory of Insect Developmental and Evolutionary Biology CAS Center for Excellence in Molecular Plant SciencesShanghai Institute of Plant Physiology and EcologyChinese Academy of SciencesShanghai 200032China CAS Center for Excellence in Biotic Interactions University of Chinese Academy of SciencesBeijing 100049China School of Life Science and Technology ShanghaiTech UniversityShanghai 201210China

Fungal G-protein coupled receptors(GPCRs)play essential roles in sensing environmental cues including host signals.The study of GPCR in mediating fungus-insect interactions is still limited.Here we report the evolution of GPCR genes encoded in the entomopathogenic Metarhizium species and found the expansion of Pth11-like GPCRs in the generalist species with a wide host range.By deletion of ten candidate genes MrGpr1–MrGpr10 selected from the six obtained subfamilies in the generalist M.robertsii,we found that each of them played a varied level of roles in mediating appressorium formation.In particular,deletion of MrGpr8 resulted in the failure of appressorium formation on different substrates and the loss of virulence during topical infection of insects but not during injection assays when compared with the wild-type(WT)strain.Further analysis revealed that disruption of MrGpr8 substantially impaired the nucleus translocation of the mitogen-activated protein kinase(MAPK)Mero-Fus3 but not the MAPK Mero-Slt2 during appressorium formation.We also found that the defect ofΔMrGpr8 could not be rescued with the addition of cyclic AMP for appressorium formation.Relative to the WT,differential expression of the selected genes have also been detected inΔMrGpr8.The results of this study may benefit the understanding of fungus-interactions mediated by GPCRs.

关键词： G-protein coupled receptor appressorium virulence signal pathway Metarhizium

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TransTM:A device-free method based on time-streaming multiscale transformer for human activity recognition

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Defence Technology（防务技术） 2024年第2期32卷 619-628页

作者： Yi Liu Weiqing Huang shang Jiang Bobai Zhao Shuai Wang Siye Wang yanfang Zhang Institute of Information Engineering Chinese Academy of Sciences School of Cyber SecurityUniversity of Chinese Academy of SciencesBeijingChina School of Information Management Beijing Information Science and Technology UniversityBeijingChina

RFID-based human activity recognition(HAR)attracts attention due to its convenience,noninvasiveness,and privacy protection.Existing RFID-based HAR methods use modeling,CNN,or LSTM to extract features effectively.Still,they have shortcomings:1)requiring complex hand-crafted data cleaning processes and 2)only addressing single-person activity recognition based on specific RF signals.To solve these problems,this paper proposes a novel device-free method based on Time-streaming Multiscale Transformer called TransTM.This model leverages the Transformer's powerful data fitting capabilities to take raw RFID RSSI data as input without pre-processing.Concretely,we propose a multiscale convolutional hybrid Transformer to capture behavioral features that recognizes singlehuman activities and human-to-human interactions.Compared with existing CNN-and LSTM-based methods,the Transformer-based method has more data fitting power,generalization,and scalability.Furthermore,using RF signals,our method achieves an excellent classification effect on human behaviorbased classification tasks.Experimental results on the actual RFID datasets show that this model achieves a high average recognition accuracy(99.1%).The dataset we collected for detecting RFID-based indoor human activities will be published.

关键词： Human activity recognition RFID Transformer

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Research at channel level on the effect of LaCl3 on Radish (Raphanus satirus L.) vacuolar membrane

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Chinese Science Bulletin 2002年第2期47卷 111-115页

作者： shang yanfang YANG Pin PEI Zhenming Institute of Molecular Science Shanxi UniversityTaiyuan 030006China Department of Biology Duke UniversityDurhamNC 27708USA

We recorded slow vacuolar (SV-type) channel currents of Radish vacuoles successfully for the first time by using the whole-vacuolar patch-clamp recording mode. SV-type currents would increase and threshold potentials of activation would shift towards more negative values with the increase of concentrations of cytosolic Ca2+. When 2.5 mmol/ L LaCl3 and 4 mmol/L EGTA were added to bath solutions, SV-type currents were suppressed remarkably. Then adding LaCl3 with different concentrations to pipette solutions, we found that LaCl3 with higher concentrations (>4 × 10-7mol/L) had a strong inhibitory effect on SV-type currents, while LaCl3 with lower concentrations (≤4 × 10-7mol/L) promoted channel currents. This promoting effect provides an impor-tant basis at channel level for researching further the effects of rare earth on physiological activities of plants and the production-increase effects of rare earth fertilizers on crops.

关键词： whole-vacuolar patch-clamp recording SV-type channel current cytosolic Ca2+ La3+.

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Unravelling the biosynthetic mechanism of oosporein in Beauveriabassiana

Unravelling the biosynthetic mechanism of oosporein in Beauv...

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中国菌物学会第六届会员代表大会（2014年学术年会）暨...

作者： PengFeng YongjiangXu YongliangXia yanfang shang Chengshu Wang Key Laboratory of Insect Developmental and Evolutionary Biology

Insect pathogenic fiingi produce a variety of insecticidally andpharmaceutically active compounds,including the polyketideoosporein produced by the insect pathogenic fungi Beauveria spp.and other fungi.Oosporein was first discoveredmore than fifty years ago,however,the genes responsible for its biosynthesiswere unknown until this study.Based onbioinformaticanalysisand targeted gene disruptions,we report the genecluster responsible for oosporein biosynthesis in the insect fungal pathogenB.bassiana.Thepolyketide synthase（PKS）OpS1 belongs to aniterative type Ⅰ PKS andis responsible for the biosynthesis of the precursor（orsellinic acid） for oosporein.Then the hydroxylase OpS3 catalyzes the decarboxylation and hydroxylation of orsellinic acid.The laccase OpS4 then oxidizes the intermediate into a quinonoid compound,which is then dimerized through a radical reaction.The biosynthesis of oosporein is enhanced by the transcription factor OpS3.Insect bioassays showed that oosporeinis important for the insect-killing process inB.bassiana and may also function as an antagonist against other saprophytic bacteria.The resultof this study will facilitate the efficient production and development of oosporein as potential bioinsecticides or pharmaceuticals.

关键词： Oosporein Beauveriabassiana polyketide synthase biosynthesis virulence,

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The response regulator Skn7 controls cell wall integrity,sporulation,autolysis and virulence in Metarhizium robertsii

The response regulator Skn7 controls cell wall integrity,spo...

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中国菌物学会第六届会员代表大会（2014年学术年会）暨...

作者： yanfang shang Wei Huang Qiang Gao Peilin Chen Yixiong Chen Chengshu Wang Key Laboratory of Insect Developmental and Evolutionary Biology Institute of Plant Physiology and EcologyShanghai Institutes for Biological SciencesChinese Academy of Sciences

Two-component signaling pathways include histidine kinases,response regulators,and sometimes histidine-containing phosphotransfer proteins.A response regulator protein of two-component system,skn7-homologous gene was identified in the insect pathogenic fungus Metarhizium robertsii and designated as Mrskn7.To study its potential function,the Mrskn7 gene deletion mutants were obtained.Under normal growth conditions（e.***.potato dextrose agar）,the mutant lost the ability to sporulate and demonstrated hyphal autolysis at late growth stage.Under the salt（KCl or NaCl） stresses,the mutant was partially rescued to produce spores,but the virulence was significantly decreased when compared to the wild type.Gene expression analysis by real-time PCR showed that the expressions of autolysis-related genes like NagA,chitinase and protease genes were significantly up-regulated in △Mrskn7.Cell wall staining results revealed a serious defect in cell wall composition in Mrskn7 mutant.Yeast one-hybrid tests confirmed that Mrskn7 could function as a transcription factor to control the expression of genes involved in regulating cell wall biosynthesis,cell autolysis and sporulation in M.robertsii.

关键词： Metarhizium robertsii Two-component system Response Regulator Cell Wall Integrity Sporulation Autolysis Virulence

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Functional studies of the genes involved in osmotic stress responses and virulence in Metarhizium robertsii

Functional studies of the genes involved in osmotic stress r...

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中国菌物学会第六届会员代表大会（2014年学术年会）暨...

作者： Peilin Chen Wei Huang yanfang shang Yixiong Chen Peng Feng Chengshu Wang Key Laboratory of Insect Developmental and Evolutionary Biology

Metarhizium spp.are ubiquitous soil-dwelling insect-pathogenic fungi in a variety of ecosystems to thrive by counteracting different stresses.We report the characterizations of a Hog1-like gene,a Ssk1-like gene and a Atf1-liks gene in M.robertsii,designated as Mrsskl,Mrhog1 and Mratf1,respectively.Our results showed that Mrssk1 and Mrhog1 were involved in fungal responses to many environmental stresses,including high osmolality,high temperature,and UV radiation.Disruption of MrSsk1 led to the failure of phosphorylation and nuclear translocation of MrHog1 in response to osmotic stress.MrAtf1 is a transcription factor and expressed by the fungus in the conidia,hyphae and appressorium of M.robertsii,but not in hyphal body.Mratf1 deletion mutant was insensitive to osmotic stress.The interaction between MrHog1 and Mratf1 was confirmed by pull down and yeast two-hybrid assay.Insect bioassays indicated that Mrsskl,Mrhog1 and Mratf1 mutants had reduced virulence when compared to the wild-type strain.These data indicated that the stress-activated MrSskl-MrHog1 pathway and MrAtfl are involved in various stress responses and functionally important for virulence in M.robertsii.

关键词： Metarhizium robertsii Mrssk1 Mrhog1 Mratf1 osmotic stress virulence

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The MrSsk1-MrHog1 Pathway is Important for Stress Responses and Virulence of Metarhizium robertsii

The MrSsk1-MrHog1 Pathway is Important for Stress Responses ...

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2014年第七届中国模式真菌研讨会

作者：陈培林 Wei Huang yanfang shang Yixiong Chen Peng Feng Chengshu Wang Key Laboratory of Insect Developmental and Evolutionary Biology Institute of Plant Physiology and EcologyShanghai Institutes for Biological SciencesChinese Academy of Sciences

Metarhizium spp.are ubiquitous soil-dwelling insect-pathogenic fungi and are found in a variety of ecosystems to counteract a wide variety stresses.Thus,it is important to understand the mechanisms involved in the responses to environmental stresses in entomopathogenic fungi.Here,we report the characterizations of a Hogl-like gene and an Sskl-like gene in M.robertsii,designated as Mrsskl and Mrhogl,respectively.Our results shown that these two genes are involved in fungal responses to many environmental stresses including high osmolarity,high temperature,and UV radiation.Disruption of MrSskl resulted in the phosphorylation failure of

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Linkage of programmed cell death control to fungal development and virulence in Metarhizium robertsii

Linkage of programmed cell death control to fungal developme...

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中国菌物学会第六届会员代表大会（2014年学术年会）暨...

作者： Yixiong Chen Zhibing Duan Wei Huang Peilin Chen yanfang shang Yongliang Xia Chengshu Wang Key Laboratory of Insect Developmental and Evolutionary Biology

Programmed cell death（PCD） is a genetically controlled and highly conserved process,classified into apoptosis and autophagy.Yeast Atg8 has been identified as one of the key autophagic proteins that is associated with the autophagosome membrane,and human Bax inhibitor 1（BI-1） is originally identified as a suppressor of apoptosis to inhibit pro-apoptotic protein Bax.Until this study,the effect of PCD on fungal biology and infection of insect pathogens remains largely unknown.Here,we report the identification and functions of MrATG8 and MrBI-1,orthologs of yeast Atg8 and humaaBI-1in the entomopathogenic fungus Metarhizium robertsii,respectively.We found that both MrATG8 and MrBI-1 play important roles in fungal development and virulence.Compared to the wild-type and gene-rescued mutant,Mratg8△ failed to form the infection-structure appressorium andthe defense response of which against insect immunity is impaired.In addition,accumulation of lipid droplets（LDs） is significantly reduced in the conidia of Mratg8△ and the pathogenicity of the mutant is dramatically impaired.We also found that the LD-specific perilipin-like protein is significantly lowered by deletion of MrATG8.MrBI-1 can partially rescue the Bax-induced growth defect in yeast and deletion of MrBI-1 impaired fungal development,heat tolerance and virulence in M.robertsii.We demonstrated that inactivation of MrBI-1 increased fungal sensitivity to farnesol（FOH）,which can trigger apoptosis-like cell death.Our data not only advance our understanding of the two forms of PCD in fungi,but also provide evidence to connect autophagy with lipid metabolisms.

关键词： Metarhizium robertsii PCD apoptosis autophagy virulence lipid metabolism heat tolerance.

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气象因子对察右中旗高山草甸草原牧草生长发育的影响

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内蒙古气象 2023年第2期 26-31页

作者：尚艳芳察右中旗气象局内蒙古察右中旗013550

利用察右中旗气象局生态监测牧草数据和气象观测资料,分析气象因子对察右中旗高山草甸草原牧草生长发育的影响。结果表明:在气候变暖的背景下,脚苔草和狐茅两种典型牧草返青期提前,开花期提前,黄枯期推迟,生长季长度延长,平均每年分别延... 详细信息

利用察右中旗气象局生态监测牧草数据和气象观测资料,分析气象因子对察右中旗高山草甸草原牧草生长发育的影响。结果表明:在气候变暖的背景下,脚苔草和狐茅两种典型牧草返青期提前,开花期提前,黄枯期推迟,生长季长度延长,平均每年分别延长3.0d和2.9d。气温和风速是影响典型牧草发育期的主要因素,气温与两种典型牧草的返青期呈极显著负相关(P<0.01),而风速呈显著正相关(P<0.05),气温与黄枯期和生长季呈极显著正相关,而风速呈显著负相关;气温与典型牧草生长高度、自然草层生物指标均呈极显著正相关,风速与其均呈极显著负相关。降水是影响牧草生长发育的又一主要因素,降水量与脚苔草开花期呈显著负相关,与牧草盖度、鲜重和干重呈正相关,但对牧草生长高度影响不大。这说明辉腾锡勒高山草甸草原水分条件基本能够满足牧草生长所需,而气温偏低、风速偏大,是限制典型牧草生长高度的主要气象因子。同时,找出各气象因素对典型牧草高度和自然草层生物指标影响的具体变化趋势,为牧草生长全程系列化服务业务提供参考依据和数据支持,提高牧业气象监测、预报、服务业务产品的定量化、精准化水平,对生态环境变化及保护利用和评价牧业气候资源开发利用具有重要的科学意义,为察右中旗旅游业发展打下坚实的基础。

关键词：气象因子察右中旗高山草甸草原牧草生长发育影响

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大果榛子在察右中旗引种实验及分析

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内蒙古气象 2022年第2期 45-48页

作者：尚艳芳王晓君察右中旗气象局内蒙古察右中旗013550 乌兰察布市气象局内蒙古乌兰察布012000

2019年从辽宁开原引进大果榛子幼苗,在乌兰察布市察右中旗开始栽培试验。经过2年的栽植,发现大果榛子幼苗栽植成活率不高,大果榛子不适宜在察右中旗引种。通过对引种地与实验地气象数据进行对比以及田间管理方面深入分析,找出气温、降... 详细信息

2019年从辽宁开原引进大果榛子幼苗,在乌兰察布市察右中旗开始栽培试验。经过2年的栽植,发现大果榛子幼苗栽植成活率不高,大果榛子不适宜在察右中旗引种。通过对引种地与实验地气象数据进行对比以及田间管理方面深入分析,找出气温、降水和相对湿度是大果榛子引种后成活率低的主要气候因素,引种不规范和养护管理简单化以及引种时间和季节不当是大果榛子引种失败的又一主要因素。大果榛子在察右中旗引种实验及分析,旨在为今后经济林果引种栽培提供参考。

关键词：察右中旗大果榛子引种实验分析

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