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aBIOTECH 2023年第2期4卷 83-96页

作者： Hao He linhao ge Zhaolei Li Xueping Zhou Fangfang Li State Key Laboratory for Biology of Plant Diseases and Insect Pests Institute of Plant ProtectionChinese Academy of Agricultural SciencesBeijing100193China State Key Laboratory of Rice Biology Institute of BiotechnologyZhejiang UniversityHangzhou310058China

Autophagy plays an active anti-viral role in *** evidence suggests that viruses can inhibit or manipulate autophagy,thereby winning the arms race between plants and ***,we demonstrate that overexpression of an m^(6)A writer from Solanum lycopersicum,SlHAKAI,could negatively regulate pepino mosaic virus(PepMV)infection,inhibit viral RNA and protein accumulations by affecting viral m^(6)A levels in tomato plants and vice *** PepMV-encoded RNA-dependent RNA polymerase(RdRP)directly interacts with SlHAKAI and reduces its protein *** RdRP-mediated decreased protein accumulation of SlHAKAI is sensitive to the autophagy inhibitor 3-methyladenine and is compromised by knocking down a core autophagy ***,PepMV RdRP could interact with an essential autophagy-related protein,***,SlHAKAI,and SlBeclin1 interaction complexes form bright granules in the *** of Beclin1 in Nicotiana benthamiana plants abolishes the RdRP-mediated degradation of SlHAKAI,indicating the requirement of Beclin1 in this *** study uncovers that the PepMV RdRP exploits the autophagy pathway by interacting with SlBeclin1 to promote the autophagic degradation of the SlHAKAI protein,thereby inhibiting the m^(6)A modification-mediated plant defense responses.

关键词： Pepino mosaic virus HAKAI Autophagy Beclin1

geminiviral C2 proteins inhibit active autophagy to facilitate virus infection by impairing the interaction of ATG7 and ATG8

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Journal of Integrative Plant Biology 2023年第5期65卷 1328-1343页

作者： Buwei Cao linhao ge Mingzhen Zhang Fangfang Li Xueping Zhou State Key Laboratory for Biology of Plant Diseases and Insect Pests Institute of Plant ProtectionChinese Academy of Agricultural SciencesBeijing 100193China State Key Laboratory of Rice Biology Institute of BiotechnologyZhejiang UniversityHangzhou 310058China

Autophagy is a conserved intracellular degradation process that plays an active role in plant response to virus *** we report that geminiviruses counteract activated autophagymediated antiviral defense in plant cells through the C2 proteins they *** found that,in Nicotiana benthamiana plants,tomato leaf curl Yunnan virus(TLCYnV)infection upregulated the transcription levels of autophagy-related genes(ATGs).Overexpression of NbATG5,NbATG7,or NbATG8a in *** plants decreased TLCYnV accumulation and attenuated viral ***,transgenic overexpression of NbATG7 promoted the growth of *** plants and enhanced plant resistance to *** further revealed that the C2 protein encoded by TLCYnV directly interacted with the ubiquitinactivating domain of *** interaction competitively disrupted the ATG7–ATG8 binding in *** and Solanum lycopersicum plants,thereby inhibiting autophagy ***,we uncovered that the C2-mediated autophagy inhibition mechanism was conserved in three other *** summary,we discovered a novel counter-defensive strategy employed by geminiviruses that enlists their C2 proteins as disrupters of ATG7–ATG8 interactions to defeat antiviral autophagy.

关键词： autophagy C2 counterdefense geminivirus plant defense the ATG7–ATG8 interaction

m^(6)A modification of plant virus enables host recognition by NMD factors in plants

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Science China(Life Sciences) 2024年第1期67卷 161-174页

作者： Hao He linhao ge Yalin Chen Siwen Zhao Zhaolei Li Xueping Zhou Fangfang Li State Key Laboratory for Biology of Plant Diseases and Insect Pests Institute of Plant ProtectionChinese Academy of Agricultural SciencesBeijing 100193China State Key Laboratory of Rice Biology Institute of BiotechnologyZhejiang UniversityHangzhou 310029China

N^(6)-methyladenosine(m^(6)A)is the most abundant eukaryotic mRNA modification and is involved in various biological *** evidence has implicated that m^(6)Amodification is an important anti-viral defense mechanism in mammals and plants,but it is largely unknown how m^(6)Aregulates viral infection in *** we report the dynamic changes and functional anatomy of m^(6)Ain Nicotiana benthamiana and Solanum lycopersicum during Pepino mosaic virus(PepMV)infection.m^(6)Amodification in the PepMV RNA genome is conserved in these two *** of the m^(6)Awriters,mRNA adenosine methylase A(MTA),and HAKAI inhibit the PepMV RNA accumulation accompanied by increased viral m^(6)Amodifications,whereas deficiency of these writers decreases the viral RNA m^(6)Alevels but enhances virus *** study reveals that the cytoplasmic YTH-domain family protein NbECT2A/2B/2C as m^(6)Areaders are involved in anti-viral ***-protein interactions indicate that NbECT2A/2B/2C interact with nonsense-mediated mRNA decay(NMD)-related proteins,including NbUPF3 and NbSMG7,but not with NbUPF1.m^(6)Amodification-mediated restriction to PepMV infection is dependent on NMD-related *** findings provide new insights into the functionality of m^(6)Aanti-viral activity and reveal a distinct immune response that NMD factors recognize the m^(6)Areaders-viral m^(6)ARNA complex for viral RNA degradation to limit virus infection in plants.

关键词： m^(6)A Pepino mosaic virus plant defense m^(6)A readers NMD factors

SUMOylation-modified Pelota-Hbs1 RNA surveillance complex restricts the infection of potyvirids in plants

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Molecular Plant 2023年第3期16卷 632-642页

作者： linhao ge Buwei Cao Rui Qiao Hongguang Cui Shaofang Li Hongying Shan Pan Gong Mingzhen Zhang Hao Li Aiming Wang Xueping Zhou Fangfang Li State Key Laboratory for Biology of Plant Diseases and Insect Pests Institute of Plant ProtectionChinese Academy of Agricultural SciencesBejingChina Key Laboratory of Green Prevention and Control of Tropical Plant Diseases and Pests Ministry of Education and College of Plant ProtectionHainan UniversityHaikouHainanChina London Research and Development Centre Agriculture and Agri-Food CanadaLondonONCanada Department of Biology Western UniversityLondonONCanada state Key Laboratory of Rice Biology Institute of BiotechnologyZhejiang UniversityHangzhouZhejiangChina

RNA quality control nonsense-mediated decay is involved in viral restriction in both plants and ***,it is not known whether two other RNA quality control pathways,nonstop decay and no-go decay,are capable of restricting viruses in ***,we show that the evolutionarily conserved Pelota–Hbs1 complex negatively regulates infection of plant viruses in the family Potyviridae(termed potyvirids),the largest group of plant RNA viruses that accounts for more than half of the viral crop damage *** enables the recognition of the functional G1-2A6-7 motif in the P3 cistron,which is conserved in almost all *** allows Pelota to target the virus and act as a viral restriction ***,Pelota interacts with the SUMO E2-conjugating enzyme SCE1 and is SUMOylated in *** Pelota SUMOylation disrupts the ability to recruit Hbs1 and inhibits viral RNA *** findings reveal the functional importance of Pelota SUMOylation during the infection of potyvirids in plants.

关键词： SUMOylation Pelota-Hbs1 G1-2A6-7 motif Potyviruses

Plant and animal positive-sense single-stranded RNA viruses encode small proteins important for viral infection in their negative-sense strand

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Molecular Plant 2023年第11期16卷 1794-1810页

作者： Pan Gong Qingtang Shen Mingzhen Zhang Rui Qiao Jing Jiang Lili Su Siwen Zhao Shuai Fu Yu Ma linhao ge Yaqin Wang Rosa Lozano-Durán Aiming Wang Fangfang Li Xueping Zhou State Key Laboratory for Biology of Plant Diseases and Insect Pests Institute of Plant ProtectionChinese Academy of Agricultural SciencesBeijingChina School of Basic Medical Sciences Fujian Medical UniversityFuzhouChina State Key Laboratory of Rice Biology and Breeding Institute of BiotechnologyZhejiang UniversityHangzhouZhejiangChina University of the Chinese Academy of Sciences BeijingChina Department of Plant Biochemistry Center for Plant Molecular Biology(ZMBP)Eberhard Karls University72076 TübingenGermany London Research and Development Center Agriculture and Agri-Food CanadaLondonONCanada

Positive-sense single-stranded RNA(+ssRNA)viruses,the most abundant viruses of eukaryotes in nature,require the synthesis of negative-sense RNA(-RNA)using their genomic(positive-sense)RNA(+RNA)as a template for *** on current evidence,viral proteins are translated via viral+RNAs,whereas-RNA is considered to be a viral replication intermediate without coding ***,we report that plant and animal+ssRNA viruses contain small open reading frames(ORFs)in their-RNA(reverse ORFs[rORFs]).Using turnip mosaic virus(TuMV)as a model for plant+ssRNA viruses,we demonstrate that small proteins encoded by rORFs display specific subcellularlocalizations,and confirm the presence of rORF2 in infected cells through mass spectrometry *** protein encoded by TuMV rORF2 forms punctuate granules that are localized in the perinuclear region and co-localized with viral replication *** rORF2 protein can directly interact with the viral RNA-dependent RNA polymerase,and mutation of rORF2 completely abolishes virus infection,whereas ectopic expression of rORF2 rescues the mutant ***,we show that several rORFs in the-RNA of severe acute respiratory syndrome coronavirus 2(SARS-CoV-2)have the ability to suppress type l interferon production and facilitate the infection of ve-sicular stomatitis *** addition,we provide evidence that TuMV might utilize internal ribosome entry sites to translate these small *** together,these findings indicate that the-RNA of+ssRNA vi-ruses can also have the coding capacity and that small proteins encoded therein play critical roles in viral infection,revealing a viral proteome larger than previously thought.

关键词： positive-sense single-stranded RNA viruses small proteins negative-sense RNA turnip mosaic virus severe acute respiratory syndrome coronavirus 2

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m^(6)A修饰与植物RNA病毒侵染

中国科学：生命科学 2024年第3期54卷 447-458页

作者：葛林豪潘福安何浩贾明轩李方方周雪平中国农业科学院植物保护研究所植物病虫害综合治理全国重点实验室北京100193 浙江大学生物技术研究所水稻生物学国家重点实验室杭州310058

N6-甲基腺嘌呤(N6-methyladenosine,m^(6)A)是真核生物m RNA中丰度最高的RNA转录后化学修饰.RNA的m^(6)A修饰主要由甲基化转移酶(writers)、去甲基化酶(erasers)以及阅读蛋白(reader proteins)共同调控.近年的研究表明,m^(6)A修饰在植物病毒侵染中发挥了重要作用,相关调控机制成为植物病毒领域的研究热点.本文概述了植物RNA m^(6)A修饰相关蛋白的基本组成和m^(6)A修饰的检测技术,重点阐述了m^(6)A修饰在植物与RNA病毒互作中的作用,并提出了今后植物RNA病毒m^(6)A修饰功能研究的方向.

关键词： m^(6)A修饰植物植物RNA病毒

采用优化的数字PCR方法分析转基因小麦外源基因拷贝数

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中国农业科学 2020年第10期53卷 1931-1939页

作者：琚鹏举宁蕾葛林豪许成杰史华伟梁凯歌马亮刘陶然陈明孙黛珍山西农业大学农学院中国农业科学院作物科学研究所中央民族大学石家庄市农林科学研究院

【目的】探索基于数字PCR的小麦基因拷贝数分析技术,提高目标基因拷贝数的分析通量,促进小麦基因组研究及基因工程研究。【方法】采用中国农业科学院作物科学研究所小麦抗逆分子育种课题组创制的高抗小麦黄花叶病转nib8小麦为试验材料,... 详细信息

【目的】探索基于数字PCR的小麦基因拷贝数分析技术,提高目标基因拷贝数的分析通量,促进小麦基因组研究及基因工程研究。【方法】采用中国农业科学院作物科学研究所小麦抗逆分子育种课题组创制的高抗小麦黄花叶病转nib8小麦为试验材料,使用小麦D基因组中的单拷贝纯合内源基因PINb-D1b(籽粒硬度基因)为内参基因,根据转化的抗病基因nib8序列设计4对特异性引物及相应探针,通过试验确定探针和引物的最优浓度,优化体系的退火温度,寻找最合适的模板浓度。然后用数字PCR检测转nib8小麦中外源基因的拷贝数;同时用Real-time PCR和Southern blot 2种不同方法分析的拷贝数结果,验证上述采用数字PCR方法分析拷贝数的准确性;以PINb-D1b内参基因检测Wx012(蜡质基因)和SSII(淀粉合成基因)2个内参基因的拷贝数,以Wx012和SSII为内参基因检测转nib8小麦中外源基因的拷贝数,验证以PINb-D1b内参基因的检测结果准确性;在nib8的不同区段设计特异性引物来检测转nib8株系拷贝数分析结果是否有差异。最终,建立基于数字PCR技术的高通量检测小麦目标基因拷贝数的分析方法。【结果】通过试验最终确定了基于数字PCR方法的小麦目标基因拷贝数的检测体系。试验确定引物和探针的最佳终浓度分别为500和250 nmol·L-1,确定体系反应最佳退火温度为59℃,最佳DNA模板量为40 ng。以PINb-D1b作为内参基因检测转nib8小麦中nib8的拷贝数,拷贝数检测结果显示第12、16、17、23、29、30株系中nib8拷贝数分别为7个、1个、1个、1个、1个、7个;同时通过比较发现此结果与Real-time PCR以及Southern blot结果一致;在转基因株系中,以PINb-D1b为内参基因,对Wx012和SSII 2个基因进行拷贝数进行分析,同时发现采用这3种内参基因分析转nib8小麦中nib8基因拷贝数的结果一致,说明这3个内参基因都适合用于数字PCR方法;在nib8上游、中游和下游不同区段的设计引物分析拷贝数检测结果一致。【结论】优化了基于数字PCR方法的小麦目标基因拷贝数分析方法和反应体系,确立了基于数字PCR方法的小麦目标基因拷贝数的检测体系,数字PCR分析结果稳定、可靠,检测通量明显提高,具有一定的应用前景。

关键词：小麦基因拷贝数数字PCR 内参基因

Improving Low Bitrate Video Coding via Computation Incorporating A Priori Information

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China Communications 2015年第9期12卷 22-34页

作者： Tao Xiaoming Dong linhao Li Shaoyang Li Yang ge Ning Lu Jianhua Department of Electronics Engineering Tsinghua University School of Aerospace Engineering Tsinghua University

The growing number of mobile users, as well as the diversification in types of services have resulted in increasing demands for wireless network bandwidth in recent years. Although evolving transmission techniques are able to enlarge the network capacity to some degree, they still cannot satisfy the requirements of mobile users. Meanwhile, following Moore's Law, the data processing capabilities of mobile user terminals are continuously improving. In this paper, we explore possible methods of trading strong computational power at wireless terminals for transmission efficiency of communications. Taking the specific scenario of wireless video conversation, we propose a model-based video coding scheme by learning the structures in multimedia contents. Benefiting from both strong computing capability and pre-learned model priors, only low-dimensional parameters need to be transmitted; and the intact multimedia contents can also be reconstructed at the receivers in real-time. Experiment results indicate that, compared to conventional video codecs, the proposed scheme significantly reduces the data rate with the aid of computational capability at wireless terminals.

关键词： computational communications a priori information bandwidth-limited wireless communications model-based video coding

拟南芥膜定位蛋白At CP2调控抗冻性的功能分析

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中国农业科技导报 2019年第9期21卷 58-68页

作者：齐欣丁庆倩苏晨徐伟亚牟永莹葛林豪徐兆师陈隽潘映红马有志陈明中国农业科学院作物科学研究所农作物基因资源与基因改良国家重大科学工程农业农村部麦类生物学与遗传育种重点实验室北京100081 西北农林科技大学农学院陕西杨凌712100

COR家族成员在调控植物响应低温胁迫过程中发挥重要作用。从拟南芥中克隆一个新的低温胁迫响应基因At CP2,其属于COR基因家族中的COR413亚家族,编码区612 bp,编码203个氨基酸,分子量22.75k Da,等电点9.44。亚细胞定位和基因表达模式分... 详细信息

COR家族成员在调控植物响应低温胁迫过程中发挥重要作用。从拟南芥中克隆一个新的低温胁迫响应基因At CP2,其属于COR基因家族中的COR413亚家族,编码区612 bp,编码203个氨基酸,分子量22.75k Da,等电点9.44。亚细胞定位和基因表达模式分析结果表明:At CP2定位在叶绿体膜上,对ABA、PEG、Na Cl和低温四种非生物胁迫均有响应。基因功能分析结果显示:功能缺失性突变体cp2冷处理后存活率明显高于野生型拟南芥(WT),相对电导率和MAD均低于WT,证明At CP2基因负向调控植物抗冻性。蛋白质组及qRT-PCR分析结果证明At CP2通过负向调控4个ABA及低温胁迫相关基因(包括ABA2、KIN1、COR47和COR15B)的表达影响植物抗冻性。总之,At CP2基因的功能分析结果对了解COR413基因家族的功能提供了参考。

关键词：拟南芥 At CP2基因低温胁迫蛋白质组下游基因

谷子MYB类转录因子SiMYB42提高转基因拟南芥低氮胁迫耐性

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遗传 2018年第4期40卷 327-338页

作者：丁庆倩王小婷胡利琴齐欣葛林豪徐伟亚徐兆师周永斌贾冠清刁现民闵东红马有志陈明中国农业科学院作物科学研究所农作物基因资源与基因改良国家重大科学工程农业部麦类生物学与遗传育种重点实验室北京100081 西北农林科技大学农学院旱区作物逆境生物学国家重点实验室杨凌712100

Myeloblastosis(MYB)类转录因子是高等植物中最大的转录因子家族之一,在植物发育及防御反应过程中发挥重要作用,还参与植物对干旱等非生物胁迫的响应。谷子(Setaria italica L.)起源于中国,具有抗旱、耐瘠薄的特性,是研究单子叶作物非... 详细信息

Myeloblastosis(MYB)类转录因子是高等植物中最大的转录因子家族之一,在植物发育及防御反应过程中发挥重要作用,还参与植物对干旱等非生物胁迫的响应。谷子(Setaria italica L.)起源于中国,具有抗旱、耐瘠薄的特性,是研究单子叶作物非生物胁迫抗性的理想材料。本研究对耐低氮胁迫谷子品种郑204经低氮处理后进行转录组分析,鉴定出一个在低氮胁迫条件下明显上调的MYB类转录因子SiMYB42。系统发育树结果表明,SiMYB42属于R2R3-MYB亚族,具有2个MYB保守域;表达模式分析显示,SiMYB42在低氮、高盐、干旱和ABA胁迫条件下表达量显著上调;亚细胞定位、quantitative real-time PCR及转录激活活性分析结果表明,SiMYB42蛋白定位于植物的细胞核和细胞膜中,主要在谷子的叶部或根部表达,具有转录激活活性;基因功能分析结果表明,在正常条件下,转SiMYB42基因拟南芥与野生型Columbia-0拟南芥(WT)无明显差异,但在低氮条件下,转SiMYB42基因拟南芥的主根长、根系表面积及鲜重均显著高于WT,结果证明SiMYB42基因可以提高转基因植物对低氮胁迫的耐性;下游基因表达分析结果显示,在转SiMYB42基因拟南芥中,参与植物氮素转运的硝酸盐转运基因NRT2.1、NRT2.4和NRT2.5的表达水平均高于WT,启动子分析结果显示NRT2.1、NRT2.4和NRT2.5基因启动子序列中均具有MYB结合位点。以上结果证明,SiMYB42可以通过调控下游硝酸盐转运体基因的表达提高植物在低氮条件下的耐性。本研究揭示了SiMYB42基因在低氮胁迫反应途径中的作用,为进一步了解谷子低氮胁迫响应的调控网络奠定了基础。

关键词：谷子(Setaria italica L.) MYB类转录因子低氮胁迫基因表达分析基因功能分析