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Quantitative analysis of the interaction of heterologous viruses with Plum pox virus in C5 HoneySweet transgenic plums

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Journal of Integrative Agriculture 2019年第10期18卷 2302-2310页

作者： Khushwant Singh Tereza Neubauerová Jiban Kumar Kundu plant Virus and Vector Interactions Group Division of Crop Protection and Plant Health Crop Research Institute

Stone fruits are an important crop in most parts of the world and are heavily challenged by several viruses including Plum pox virus (PPV), Prune dwarf virus (PDV), Prunus necrotic ringspot virus (PNRSV), and Apple chlorotic leaf spot virus (ACLSV). We validated the PPV resistance in C5 plum plants (commercially known as HoneySweet) grown in the Czech Republic for more than 16 years in a field trial experiment under natural environmental conditions. We quantified single (PPV-Rec) and mixed viruses (PPV-Rec+ACLSV, PPV-Rec+PDV and PPV-Rec+ACLSV+PDV) in C5 transgenic plums inoculated for the period 2016 to 2018. The accumulation of PPV-Rec was high (~5.43E+05 copies) compared with that of ACLSV (~8.70E+04 copies) in the inoculated graft of C5 transgenic plants. Leaves close to the inoculum sources showed a differential level of virus titre in single and mixed infections (~10 to ~5×10^2 copies). C5 plants with permanent virus pressure showed 103- to 105-fold fewer copies of viruses than those of the inoculated graft. We observed high accumulation of conserved miRNAs such as miR167, miR69 and miR396 in C5 plants co-infected with PPV, ACLSV and PDV that are associated with its resistance against viruses. Overall, i) C5 transgenic plums showed high resistance to PPV infection, and a low level (~32 copies) of PPV only accumulated in some grafted plants, ii) high accumulation of PPV was found in inoculated grafts in single PPV infection and mixed infections, iii) heterologous virus infection sustained by ACLSV or PDV did not suppress PPV resistance, and iv) high and low conserved microRNAs accumulated in C5 plants.

关键词： PPV PDV ACLSV C5 HoneySweet resistance

Morphological and molecular characterizations of cereal cyst nematode Heterodera avenae Wollenweber, 1924 from the Czech Republic

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Journal of Integrative Agriculture 2017年第3期16卷 532-539页

作者： Shesh Kumari division of crop Protection and plant health Crop Research Institute Prague 16106 Czech Republic

The cereal cyst nematode,Heterodera avenae Wollenweber,1924,is a major pest of cereal crops throughout the world and causes serious yield losses,especially of *** studies have shown that this species is widely distributed in the Czech *** this study,seven populations of *** were molecularly studied,and one population was morphologically *** regions（18S,28S,and internal transcribed spacer 1）of ribosomal DNA were sequenced and the analysis of the 18S gene of six populations did not reveal any variation,whereas the internal transcribed spacer 1and 28S sequences of six populations differed by only two nucleotides from a population in？*** and quick identification of cereal cyst nematodes is important for effective control measures and ribosomal sequence analyses of seven populations in this study will be useful in future phylogenetic studies of Heterodera *** in the Czech Republic.

关键词： Heterodera avenae PCR ribosomal DNA sequencing nematode

Wheat streak mosaic virus： incidence in field crops, potential reservoir within grass species and uptake in winter wheat cultivars

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Journal of Integrative Agriculture 2017年第3期16卷 523-531页

作者： Jana Chalupniková Jiban Kumar Kundu Khushwant Singh Pavla Bartaková Eva Beoni division of crop Protection and plant health Crop Research Institute Prague 16106 Czech Republic Department of Experimental plant Biology Faculty of Science Charles University in Prague Prague 12843 Czech Republic

Wheat streak mosaic virus（WSMV）has become a re-emerging pathogen in cereal crops in the Czech *** was first reported in the former Czechoslovakia in the early 1980s,and then no record of the virus was documented until *** incidence of the virus was recorded in recent years in several winter wheat fields and many grass ***,we surveyed the incidence of WSMV in cereal *** results demonstrated the existence of the virus in winter wheat and volunteer wheat during each year of the monitoring period,which spanned from 2013–*** the range of infected samples was low（6.4%of the total tested samples）,a high incidence of well-distributed virus was *** at least six fields,the virus reached severe and potentially epidemic *** accordance with our previous report detailing WSMV infection of native grasses,we tested several grass species commonly grown in the Czech *** found that some grass species acted as experimental hosts and possible reservoirs of the virus;these included Anthoxanthum odoratum（sweet vernal grass）,Arrhenatherum elatius（false oat-grass）,Lolium multiflorum（Italian rye-grass）,Bromus japonicus（Japanese chess）,Echinochloa crus-galli（barnyard grass）,Holcus lanatus（meadow soft grass）and Holcus mollis（creeping soft grass）.Some of these grass species are also important weeds of cereals,which may be the potential source of WSMV infection in cereal *** widely used winter wheat cultivars were tested in the field after artificial inoculation with WSMV to evaluate virus titre by ***,the tested cultivars had a low virus titre,which is associated with mild disease symptoms and may provide a good level of crop resistance to WSMV.

关键词： WSMV survey grass species cereal crops

Molecular diversity of barley yellow dwarf virus-PAV from China and the Czech Republic

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Journal of Integrative Agriculture 2020年第11期19卷 2736-2745页

作者： May Oo KHINE Brozenka MICHAELA LIU Yan Jiban Kumar KUNDU WANG Xi-feng State Key Laboratory for Biology of plant Diseases and Insect Pests Institute of Plant ProtectionChinese Academy of Agricultural SciencesBeijing 100193P.R.China division of crop Protection and plant health Crop Research InstitutePraha 16106Czech Republic

Wheat yellow dwarf disease(BYD),caused by different species of barley/cereal yellow dwarf viruses(B/CYDVs),is one of the most serious cereal diseases in China and the Czech *** genetic diversity of the virus directly influences disease epidemiology,the molecular diversity and population structure of 24 Chinese isolates and 16 the Czech Republic isolates of BYDV-PAV from different regions in two countries were analyzed by sequencing their coat protein(CP)and readthrough protein(RTP)domain(RTD)genes and comparing the sequences with six CP and 16 RTP sequences of BYDVPAV isolates from the NCBI database based on nucleotide identity position,phylogenetic analysis and nucleotide *** identities between the Chinese and the Czech Republic isolates for the CP were 76.6–99.4%,73.9–89.1%for RTD(ORF5),*** Chinese and the other country isolates showed 74.7–99.2%nucleotide identity for RTP(ORF3+ORF5).Phylogenetic analysis of CP sequences showed that 20 Chinese isolates clustered in the same clade,but the other four Chinese isolates clustered in another clade with the isolates from the Czech Republic and other *** population of BYDV-PAV in China had greater nucleotide variability and was more divergent than that in the Czech *** and ecological factors but not hosts might contribute to the population differences in the two countries.

关键词： barley yellow dwarf virus-PAV coat protein(CP) readthrough protein(RTP) phylogeny genetic diversity China the Czech Republic

DEVELOPMENT OF COMMON WHEAT GERM-PLASM RESISTANT TO BARLEY YELLOW DWARF VIRUS BY BIOTECHNOLOGY

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Science China Chemistry 1991年第9期34卷 1055-1062页

作者：辛志勇徐惠君陈孝林志珊周广和钱幼亭成卓敏 P. J. LARKIN P. BANKS R. APPELS B. GLARKE AND R. I. S. BRETTELL Institute of crop Breeding and Cultivation GAAS Institute of plant Protection GAAS CSIRO Division of Plant IndustryAustralia

Barley yellow dwarf virus (BYDV) is one of the most serious wheat diseases in China. So far no resistance has been described in common wheat. A certain level of BYDV resistance was found in thirteen Triticeae species. Thinopyrum intermedium, two octoploids derived from TH. intermedium/wheat, Zhong 4 awnless and TAF46, and one disomic addition line, L1 derived from TAF46, showed good resistance to BYDV by enzyme linked immunosorbent assay (ELISA). Two wheat/TA. intermedium translocation lines, CPI 119880 and CPI 119899, showing good BYDV resistance were developed from L1 by using both CSph mutant and tissue culture. It is found that their BYDV resistance was controlled by a single dominant gene. Two cDNA probes pEleAcc3 and pPJN8 (E1-T1) were screened for detecting Th. intermedium DNA in wheat background. A specific band for the DNA of Th. intermedium and its derivatives was found in Southern hybridization. It is also possible to determine the size of the alien segment by comparing the relative density of the specific band. Therefore, this can be used as a marker to identify the BYDV resistance in wheat breeding program.

关键词： wheat yellow dwarf virus barley yellow dwarf virus Thinopyrum intermedium addition line translocation line cDNA probe.

Two mutations in the truncated Rep gene RBR domain delayed the Wheat dwarf virus infection in transgenic barley plants

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Journal of Integrative Agriculture 2018年第11期17卷 2492-2500页

作者： Pavel Cejnar Ludmila Ohnoutkova Jan Ripl Tomas Vlcko Jiban Kumar Kundu division of crop Protection and plant health Crop Research Institute Prague 16106 Czech Republic Department of Chemical Biology and Genetics Centre of the Region Hana for Biotechnological and Agricultural Research Palack~/ University Olomouc 78371 Czech Republic

Wheat dwarf virus （WDV）, an important cereal pathogen, is closely related to Maize streak virus （MSV）, a model virus of the Mastrevirus genus. Based on its similarity to known MSV resistance strategies, a truncated part of the WDV replication- associated （RepA） gene （WDVRepA215） and the WDV RepA gene with a mutated retinoblastoma-related protein （RBR） interaction domain （WDVRepA215RBRre^t） were cloned into the plPKb002 expression vector and transformed into immature embryos of spring barley cv. Golden Promise plants through Agrobacterium-mediated transformation. A detailed study of T1-generation plants infected by leafhoppers （Psammotettix alienus） fed on infection sources of variable strength was performed over a 5-week period encompassing the initial stages of virus infection. A DNA WDV TaqMan qPCR assay normalized using the DNA puroindoline-b SYBR Green qPCR assay for samples on a per week basis revealed an approximately 2-week delay in WDVRepA215RBR^mut plants to WDVRepA215 plants before significant increases in the WDV viral levels occurred. Both WDVRepA215 and WDVRepA215RBR^mut plants showed similar levels of transgenic transcripts over the screened period; however, the transgenic plants also showed increased numbers of infected plants compared to the control plants.

关键词： Wheat dwarf virus (WDV) truncated Rep gene RBR domain qPCR screening

Improving genomic tools for outcrossing crops

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Molecular plant 2021年第4期14卷 538-540页

作者： Cathy Jubin Stefanie Griebel Timothy Beissinger division of plant Breeding Methodology&Center for Integrated Breeding Research Department of Crop ScienceUniversity of Gottingen37075

For decades,practical limitations and cost have created a bias in plant genetic studies toward species that tolerate self *** important work by Chen et al.(2021)in this issue provides key tools to overcome this bias a... 详细信息

关键词： crops breeding overcome

Virome identification in wheat in the Czech Republic using small RNA deep sequencing

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Journal of Integrative Agriculture 2020年第7期19卷 1825-1833页

作者： Khushwant SINGH Jana JAROSOVA Jan FOUSEK CHEN Huan Jiban Kumar KUNDU plant Virus and Vector Interactions Group Division of Crop Protection and Plant HeatthCrop Research InstitutePrague 16106Czech Republic Zhejiang Tianke High-Technology Development Co. Ltd.Hangzhou 310012P.R.China

High-throughput deep-sequencing technology and bioinformatics analysis of the small RNA(sRNA)population isolated from plants allows universal virus detection and complete virome reconstruction for a given *** the present sRNA deep-sequencing analysis of virus-infected wheat samples in the Czech Republic,samples were firstly tested for barley yellow dwarf viruses(BYDVs),wheat streak mosaic virus(WSMV)and wheat dwarf virus(WDV)using ELISA,RT-PCR and *** sRNA sequencing of these samples yielded more than^60 million single-end 50-bp reads with high confidence for nine field samples of ***,16.5%of reads were virus-specific and 83.5%were mapped to the *** 21-nt reads(-7.7E+06 reads)were found than 24-nt(~6.20E+06 reads)or 22-nt(-4.30E+06 reads)*** novo assembly of the high-quality contigs revealed the presence of three earlier repoted viruses in the Czech Republic:BYDVs(31.48%),WSMV(24.23%)and WDV(2666%).We also showed the presence of cereal yellow dwarf virus(14.33%;two species CYDV-RPS and CYDV-RPV family Luteoviridae/Polerovirus)and wheat yellow dwarf virus(WYDV,3.30%;Luteoviridae).Phylogenetic analysis showed CYDV and WYDV grouped separately from ***,several recombination breakpoints were found among the groups of yellow dwarf viruses(BYDVs,CYDV,and WYDV).Using RNA deep sequencing,we confirmed the presence of the three known viruses(BYDVs,WSMV,and WDV)and the first record of two species of CYDV and WYDV in wheat in the Czech Republic.

关键词： NGS sRNA BYDVs CYDV WSMV WYDV WDV

Quantifying relationships between rooting traits and water uptake under drought in Mediterranean barley and durum wheat

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Journal of Integrative plant Biology 2014年第5期56卷 455-469页

作者： Pedro Carvalho Sayed Azam-Ali M.John Foulkes division of plant and crop Sciences University of Nottingham Sutton Bonington Campus The University of Nottingham Malaysia Campus Jalan Broga Semenyih Selangor Darul

In Mediterranean regions drought is the major factor limiting spring barley and durum wheat grain yields. This study aimed to compare spring barley and durum wheat root and shoot responses to drought and quantify relationships between root traits and water uptake under terminal *** spring barley（Hordeum vulgare L. cv. Rum） and two durum wheat Mediterranean cultivars（Triticum turgidum L. var durum cvs Hourani and Karim） were examined in soil‐column experiments under well watered and drought conditions. Root system architecture traits, water uptake, and plant growth were measured. Barley aerial biomass and grain yields were higher than for durum wheat cultivars in well watered conditions. Drought decreased grain yield more for barley（47%） than durum wheat（30%, Hourani）. Root‐to‐shoot dry matter ratio increased for durum wheat under drought but not for barley, and root weight increased for wheat in response todrought but decreased for barley. The critical root length density（RLD） and root volume density（RVD） for 90% available water capture for wheat were similar to（cv. Hourani） or lower than（cv. Karim） for barley depending on wheat cultivar. For both species, RVD accounted for a slightly higher proportion of phenotypic variation in water uptake under drought than RLD.

关键词： Barley Mediterranean root architecture root length density root traits root volume density water wheat