Searching for plant NLR immune receptors conferring resistance to potyviruses

To fight against invasion by pathogens,plants have evolved an elaborate innate immune system,of which the nucleotide-binding domain leucine-rich repeat-containing receptor(NLR)acts as the sensor and immune executor.Potyviruses,comprising one of the largest genera of plant viruses,cause severe crop yield losses worldwide.Inherited crop resistance to potyviruses can be used in breeding and plant transgenesis to control disease development.This review summarizes achievements in mapping and cloning NLR genes conferring dominant resistance against potyvirus in the families Fabaceae,Solanaceae,Brassicaceae,and Cucurbitaceae.It compares mechanisms of potyviral protein recognition and downstream signaling employed by NLRs and discusses strategies for exploiting NLRs to better control diseases caused by potyviruses.

Genetic Diversity of Chinese Soybean mosaic virus Strains and Their Relationships with Other Plant Potyviruses Based on P3 Gene Sequences

Soybean mosaic virus （SMV）, a member of the genus Potyvirus, is a major pathogen of soybean plants in China, and 16 SMV strains have been identified nationwide based on a former detailed SMV classification system. As the P3 gene is thought to be involved in viral replication, systemic infection, pathogenicity, and overcoming resistance, knowledge of the P3 gene sequences of SMV and other potyviruses would be useful in efforts to know the genetic relationships among them and control the disease. P3 gene sequences were obtained from representative isolates of the above-mentioned 16 SMV strains and were compared with other SMV strains and 16 Potyvirus species from the National Center for Biotechnology GenBank database. The P3 genes from the 16 SMV isolates are composed of 1041 nucleotides, encoding 347 amino acids, and share 90.7-100% nucleotide （NT） sequence identities and 95.1-100% amino acid （AA） sequence identities. The P3 coding regions of the 16 SMV isolates share high identities （92.4-98.9% NT and 96.0-100% AA） with the reported Korean isolates, followed by the USA isolates （88.5-97.9% NT and 91.4-98.6% AA）, and share low identities （80.5-85.2% NT and 82.1-84.7% AA） with the reported HZ 1 and P isolates from Pinellia ternata. The sequence identities of the P3 genes between SMV and the 16 potyviruses varied from 44.4 to 81.9% in the NT sequences and from 21.4 to 85.3% in the AA sequences, respectively. Among them, SMV was closely related to Watermelon mosaic virus （WMV）, with 76.0-81.9% NT and 77.5-85.3% AA identities. In addition, the SMV isolates and potyvirus species were clustered into six distinct groups. All the SMV strains isolated from soybean were clustered in Group I, and the remaining species were clustered in other groups. A multiple sequence alignment analysis of the C-terminal regions indicated that the P3 genes within a species were highly conserved, whereas those among species were relatively variable.

Molecular Characterization of a Large Diversity of Bambara Groundnut Potyviruses (<i>Vigna subterranea</i>(L.) Verdc.) in Burkina Faso

Potyviruses are major constraints to grain legume production by causing significant yield losses. Potyviruses infecting Bambara groundnut (Vigna subterranea) were investigated in Burkina Faso. Leaf samples collected from three agroclimatic zones were subjected to RT-PCR and sequence analyses. Of a total of 135 samples, 36 (26.67%) were detected positive in RT-PCR tests using potyvirus universal primers. Analysis of full coat protein (cp) sequences from 24 isolates revealed the occurrence of three groups of Bambara groundnut-infecting potyviruses. Virus isolates in group 1 shared 94.5% - 100% nucleotide (nt) identity with CABMV whereas those in group 2 and group 3 were distantly related Bean common necrosis virus (BCMNV) and Passiflora virus Ugandan which were their respective closest potyviruses. Group 2 shared 77.1% nt and 78.8% - 79.9% aa identity with BCMNV and group 3 shared 77.3% - 78.3% nt and 80.7% - 81.5% aa identity with Passiflora virus Ugandan. All three groups were confirmed by phylogenetic analyses. Taking into account potyvirus demarcation criteria, group 1 isolates belonged to CABMV species. Group 2 and group 3 were assigned to a potentially new potyviruses species and designated Bambara groundnut potyvirus 1 (BGPV1) and Bambara groundnut potyvirus 2 (BGPV2).

Characterization of volatile organic compounds in grafted tomato plants upon potyvirus necrotic infection

A headspace solid-phase microextraction-gas chromatography-mass spectrometry(HS-SPME/GC-MS)method was used to study the volatile organic compounds(VOCs)associated with the differential immune response of tomato plants infected with the recombinant strain of potato virus Y(PVY^(C)-to),necrogenic to tomato.Analysis was carried out in UC82(UC),a virus susceptible tomato variety,comparing the same UC plants grafted or not onto a virus tolerant tomato ecotype,Manduria(Ma);the three types of samples used for the GC-MS analysis were mock-inoculated UC/Ma plants,UC/Ma+PVY^(C)-to and UC+PVY^(C)-to plants;the VOCs obtained were 111.Results from symptomatic PVY^(C)-to-infected UC plants showed a VOCs composition enriched in alcohols,fatty acid derivates,benzenoids,and salicylic acid derivatives,while in mock-inoculated UC/Ma plants VOCs were mainly characterized by methyl ester compounds.The VOC profile was in line with RNAseq data analyses,denoting that PVY^(C)-to viral RNA accumulation and disease symptoms induce the specific transcriptional activation of genes involved in VOCs biosynthesis.Furthermore,principal component analysis highlighted that VOCs of PVY^(C)-to-infected and mock-inoculated grafted plants were much closer each other than that of symptomatic PVY^(C)-to-infected non-grafted UC plants.These results suggest that VOCs profiles of tomato plants are related to the viral RNA accumulation,disease intensity and graft-derived tolerance to PVY^(C)-to infection.

Potyvirus属成员基因组全序列的简并引物PCR和RACE扩增方法

Based on multi-alignment of complete polyprotein amino acid sequences of genus Potyvirus,five degenerated primers were designedThey were Sprimer(5′-GGX AAY AAY AGY GGX CAZ CC-3′),pNIa(+)(5′-TNY TGG AAM CAY TGG AT-3′),pCI2(+)(5′-GCX ACX AAX ATX ATX GAX AA-3′),pCI1(+)(5′-GTX GGX TCX GGX AAX TCX AC-3′)and pHC(+)(5′-TGY GAY AAY CAZ TTX GA-3′)(X=A,T,C or G:Y=T or C;Z=A or G;N=A or T;M=A,T or G)Using degenerated PCR and modified RACE methods,a protocol for determination of complete genome sequence of potyviruses was established and proved to be successful on five

Small RNA deep sequencing reveals the presence of multiple viral infections in cucurbit crops in Guangdong,China

Viral diseases are among the most critical damaging factors that impose a global threat to the cucurbit industry.China is the world’s leading country for the production and consumption of cucurbits.Guangdong,a province in southern China dominated by the tropical and subtropical climate,favors the survival of different plant viruses and their vectors.Five main cucurbit crops showing various disease symptoms were surveyed and collected to identify viruses infecting cucurbits in Guangdong during 2018–2020.In the field,the incidence ranged from 5-30%,or even 60-100% in the case of severely infected cucurbits.A total of 357 symptomatic samples were collected and subsequently screened for cucurbit viruses by small RNA deep sequencing and assembly(sRSA).Seventeen virus species belonging to 10 genera were identified in the five main cucurbit crops.The most common viruses were papaya ringspot virus(PRSV;Potyvirus),zucchini tigre mosaic virus(ZTMV;Potyvirus),zucchini yellow mosaic virus(ZYMV;Potyvirus),and watermelon silver mottle virus(WSMoV;Orthotospovirus),with infection rates of 24.4,19.0,17.1,and 14.3%,respectively.Notably,the most prevalent viruses were melon yellow spot orthotospovirus(MYSV)in cucumber,PRSV in squash,cucumber green mottle mosaic virus(CGMMV;Tobamovirus)in bottle gourd,WSMoV in white gourd,and ZYMV in luffa.Mixed infections were prevalent,and the types of mixed infections varied substantially in different cucurbit crops.Moreover,the full-length nucleotide sequences of watermelon green mottle mosaic virus(WGMMV),CGMMV,and watermelon virus A(WVA;Wamavirus)identified in bottle gourd were cloned and analyzed.This study is the first reporting WGMMV infecting bottle gourd in China mainland.In summary,the results demonstrate that in Guangdong,the most prevalent viruses belong to potyviruses,orthotospoviruses,and tobamoviruses groups.The findings will facilitate agricultural researchers and farmers to plan and implement effective disease control strategies aiming at timely detection and management of cucurbit-infecting viral pathogens.

云南烟草抗感PVY品种上Potyvirus属病毒种类调查

【目的】为云南省烟草抗PVY品种布局和病害防控等提供依据。【方法】2017—2019年在云南省5个市共13个植烟县(区)采集497个表现Potyvirus属病毒典型症状的烟草样品,并分析抗/感PVY品种上Potyvirus属病毒的种类。【结果】497个样品中Potyvirus属病毒检出率85.1%,烟草脉带花叶病毒(Tobacco vein banding mosaic virus,TVBMV)、红辣椒脉斑驳病毒(Chilli veinal mottle virus,ChiVMV)、马铃薯Y病毒(Potato virus Y,PVY)和烟草蚀纹病毒(Tobacco etch virus,TEV)的检出率分别为58.8%、23.3%、14.9%和7.8%。感PVY品种上Potyvirus属病毒检出率地区间差异较大,曲靖样品中TVBMV的检出率最高,玉溪样品中ChiVMV检出率最高,昭通样品中PVY检出率最高。在Potyvirus属病毒典型症状样品中,抗PVY品种比感PVY品种的PVY检出率低16.7%,抗PVY品种表现出良好的病害防治效果。【结论】昭阳区和澄江品种园的样品检测结果均证明,抗PVY品种的TVBMV检出率升高,是由于PVY发病率降低导致的,抗PVY品种不会导致TVBMV和ChiVMV危害上升。PVY高发病区可通过种植抗PVY品种来降低损失,种植前宜进行病原种类鉴定,以便将va品种布局在PVY为主的区域。云南主要烟区TVBMV和ChiVMV的发病率高,缺乏抗病品种,需开展抗TVBMV和ChiVMV的种质资源鉴定、发掘和创制。

侵染甘薯的马铃薯Y属病毒及其分子生物学研究进展

对甘薯羽状斑驳病毒、甘薯病毒Y、甘薯病毒G、甘薯潜隐病毒、甘薯轻型斑点病毒、甘薯脉花叶病毒6种侵染甘薯的马铃薯Y属(Potyvirus)病毒及其分子生物学进行了综述,并对甘薯病毒的鉴定及甘薯脱毒种苗的检测进行了展望.

西瓜花叶病毒辽宁臭椿分离物的鉴定与全序列分析

2015年10月,在辽宁省盖州市采集到表现花叶的臭椿样品。应用电子显微镜负染色技术检测到线状病毒粒子(800 nm×18 nm)。利用Potyvirus通用引物PVY-Legpoty-F/PVY-Legpoty-R对三株代表样品进行RTPCR扩增,得到750 bp阳性片段。经Blast搜索,与西瓜花叶病毒(Watermelon mosaic virus,WMV)高度同源。利用WMV-5NF/WMV-5NR、WMV-MNF/WMV-MNR和WMV-3NF/WMV-3NR三对引物扩增WMV全基因组,确定为10070核苷酸(MF418043)。基于cp基因进行同源性分析和系统进化关系分析,发现盖州臭椿分离物与北京臭椿分离物的关系最近。这是辽宁臭椿上WMV分子特征的首次报道。

菜豆普通花叶病毒辽宁分离物的鉴定与序列分析

2016年8月,于辽宁省盖州市采集到表现为黄化的菜豆样品。利用Potyvirus通用引物Legpoty-F/Legpoty-R对3个代表样品进行RT-PCR扩增,得到预期725 bp的片段。Blast显示它们与菜豆普通花叶病毒(Bean common mosaic virus,BCMV)同源性最高。利用报道的特异引物BCMV-CP-F/BCMV-CP-R扩增到约1.2 kb的片段,包含完整的外壳蛋白(coat protein,cp)。基于cp基因进行同源性分析,发现它们与美国菜豆分离物BCMV-[USA:Iowa:Phaseolus vulgaris:11](KM023744)同源性最高,为97.9%。根据Potyvirus的分类标准,认为它们是BCMV的辽宁分离物。这是辽宁首次报道菜豆感染BCMY。

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

RNA quality control nonsense-mediated decay is involved in viral restriction in both plants and animals.However,it is not known whether two other RNA quality control pathways,nonstop decay and no-go decay,are capable of restricting viruses in plants.Here,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 worldwide.Pelota enables the recognition of the functional G1-2A6-7 motif in the P3 cistron,which is conserved in almost all potyvirids.This allows Pelota to target the virus and act as a viral restriction factor.Furthermore,Pelota interacts with the SUMO E2-conjugating enzyme SCE1 and is SUMOylated in planta.Blocking Pelota SUMOylation disrupts the ability to recruit Hbs1 and inhibits viral RNA degradation.These findings reveal the functional importance of Pelota SUMOylation during the infection of potyvirids in plants.

华南三省(自治区)部分西番莲种植区花叶病病原检测及EAPV多聚蛋白基因序列分析

西番莲(Passiflora edulis Sims.)是热带亚热带地区重要经济作物,我国福建、广东、广西、海南等地广泛种植[1]。调查发现,我国南方多地西番莲花叶病为害严重,症状表现为叶片花叶、黄化、斑驳、畸形,甚至整株褪绿,严重影响西番莲的产量和品质。

马铃薯Y病毒属病毒P3和P3-PiPo蛋白功能研究进展

Potyvirus属于马铃薯Y病毒科Potyviridae,是最大的植物病毒属,给农业生产造成严重的经济损失。P3是Potyvirus属病毒中变异很大、功能较复杂的编码蛋白,涉及到病毒复制、侵染、抗性及细胞间运动;P3-PiPo是P3编码框内新近发现的Potyvirus重要编码蛋白,已证实它在病毒的细胞间运动中起着决定性的作用。对病毒蛋白功能的研究为该属病毒的研究发展提供重要的理论基础,对Potyvirus侵染机制及抗病机理研究具有指导价值。

Abscisic Acid Connects Phytohormone Signaling with RNA Metabolic Pathways and Promotes an Antiviral Response that Is Evaded by a Self- Controlled RNA Virus

A complex network of cellular receptors,RNA targeting pathways,and small-molecule signaling provides robust plant immunity and tolerance to viruses.To maximize their fitness,viruses must evolve control mechanisms to balance host immune evasion and plant-damaging effects.The genus Potyvirus comprises plant viruses characterized by RNA genomes that encode large polyproteins led by the P1 protease.A P1 autoinhibitory domain controls polyprotein processing,the release of a downstream functional RNAsilencing suppressor,and viral replication.Here,we show that P1Pro,a plum pox virus clone that lacks the P1 autoinhibitory domain,triggers complex reprogramming of the host transcriptome and high levels of abscisic acid(ABA)accumulation.A meta-analysis highlighted ABA connections with host pathways known to control RNA stability,turnover,maturation,and translation.Transcriptomic changes triggered by P1Pro infection or ABA showed similarities in host RNA abundance and diversity.Genetic and hormone treatment assays showed that ABA promotes plant resistance to potyviral infection.Finally,quantitative mathematical modeling of viral replication in the presence of defense pathways supported self-control of polyprotein processing kinetics as a viral mechanism that attenuates the magnitude of the host antiviral response.Overall,our findings indicate that ABA is an active player in plant antiviral immunity,which is nonetheless evaded by a self-controlled RNA virus.

Fusarivirus accessory helicases present an evolutionary link for viruses infecting plants and fungi

A significant number of mycoviruses have been identified that are related to plant viruses,but their evolutionary relationships are largely unexplored.A fusarivirus,Rhizoctonia solani fusarivirus 4(RsFV4),was identified in phytopathogenic fungus Rhizoctonia solani(R.solani)strain XY74 co-infected by an alphaendornavirus.RsFV4 had a genome of 10,833 nt(excluding the poly-A tail),and consisted of four non-overlapping open reading frames(ORFs).ORF1 encodes an 825 aa protein containing a conserved helicase domain(Hel1).ORF3 encodes 1550 aa protein with two conserved domains,namely an RNA-dependent RNA polymerase(RdRp)and another helicase(Hel2).The ORF2 and ORF4 likely encode two hypothetical proteins(520 and 542 aa)with unknown functions.The phylogenetic analysis based on Hel2 and RdRp suggest that RsFV4 was positioned within the fusarivirus group,but formed an independent branch with three previously reported fusariviruses of R.solani.Notably,the Hel1 and its relatives were phylogenetically closer to helicases of potyviruses and hypoviruses than fusariviruses,suggesting fusarivirus Hel1 formed an evolutionary link between these three virus groups.This finding provides evidence of the occurrence of a horizontal gene transfer or recombination event between mycoviruses and plant viruses or between mycoviruses.Our findings are likely to enhance the understanding of virus evolution and diversity.