Heterodera filipjevi continues to be a major threat to wheat production worldwide.Rapid detection and quantification of cyst nematodes are essential for more effective control against this nematode disease.In the pres...Heterodera filipjevi continues to be a major threat to wheat production worldwide.Rapid detection and quantification of cyst nematodes are essential for more effective control against this nematode disease.In the present study,a TaqManminor groove binder(TaqMan-MGB)probe-based fluorescence quantitative real-time PCR(qPCR)was successfully developed and used for quantifying H.filipjevi from DNA extracts of soil.The primers and probe designed from the obtained RAPD-SCAR marker fragments of H.filipjevi showed high specificity to H.filipjevi using DNA from isolatesconfirmed species of 23 Heterodera spp.,1 Globodera spp.and 3 Pratylenchus spp.The qPCR assay is highly sensitive and provides improved H.filipjevi detection sensitivity of as low as 4^(-3) single second-stage juvenile(J2)DNAs,10^(-3) female DNAs,and 0.01μgμL^(-1) genomic DNAs.A standard curve relating to the threshold cycle and log values of nematode numbers was generated and validated from artificially infested soils and was used to quantify H.filipjevi in naturally infested field soils.There was a high correlation between the H.filipjevi numbers estimated from 32 naturally infested field soils by both conventional methods and the numbers quantified using the qPCR assay.qPCR potentially provides a useful platform for the efficient detection and quantification of H.filipjevi directly from field soils and to quantify this species directly from DNA extracts of field soils.展开更多
The cereal cyst nematode（CCN, Heteroder aavenae） causes serious yield loss on cereal crops, especially wheat, worldwide. Daxing population in Beijing City and Huangyuan population in Qinghai Province, China, are two...The cereal cyst nematode（CCN, Heteroder aavenae） causes serious yield loss on cereal crops, especially wheat, worldwide. Daxing population in Beijing City and Huangyuan population in Qinghai Province, China, are two CCN populations. In this study, the CCN pathotypes of Daxing and Huangyuan populations were characterized by tests on 23 standard ＂International Test Assortment＂ with the local species Wenmai 19 as the susceptible control. Tested materials were grouped by three nematode populations＇ virulence on resistant genes（Rha1, Rha2, Rha3, Cre1） and nonresistant genes, varieties and lines. Both Daxing and Huangyuan populations were avirulent to Ortolan（Ha1）. Barley cvs. Ortolan, Siri, Morocco, Bajo Aragon 1-1, and Martin 403-2 were all resistant to both populations. Cultivars Herta, Harlan 43 and wheat Iskamish-K-2-light were all susceptible to Huangyuan population, all of them, however, were resistant to Daxing population. The other five oats were all resistant to the two tested CCN populations. Except Iskamisch K-2-light, all the other wheat cultivars（Capa, Loros×Koga, AUS 10894, and Psathias） were susceptible to Daxing population. Because the pathotypes of the two tested CCN populations in Beijing and Qinghai were not identical to any of the 13 pathotypes previously characterized by the test assortment, we classified Daxing and Huangyuan populations as the new pathotypes, named Ha91.展开更多
The cereal cyst nematodes(Heterodera avenae,Heterodera filipjevi,Heterodera latipons)are considered to be one of the most important plant parasitic nematodes attacking most cereals and can cause significant crop losse...The cereal cyst nematodes(Heterodera avenae,Heterodera filipjevi,Heterodera latipons)are considered to be one of the most important plant parasitic nematodes attacking most cereals and can cause significant crop losses(Sikora 1988).In China,H.filipjevi(Madzhidov 1981)Stelter,1984,was first reported from Henan province(peng et al.2010)and a few years later in Anhui province and Xinjiang Uygur Autonomous Region(peng et al.2016,2018).In December 2017,a survey for cereal展开更多
Cereal cyst nematode（Heterodera avenae, CCN） distributes worldwide and has caused severe damage to cereal crops, and a model host will greatly aid in the study of this nematode. In this research, we assessed the sen...Cereal cyst nematode（Heterodera avenae, CCN） distributes worldwide and has caused severe damage to cereal crops, and a model host will greatly aid in the study of this nematode. In this research, we assessed the sensitivity of 25 inbred lines of Brachypodium distachyon to H. avenae from Beijing, China. All lines of B. distachyon were infested by secondstage juveniles（J2s） of H. avenae from Daxing District of Beijing population, but only 13 inbred lines reproduced 0.2–3 cysts/plant, showing resistance. The entire root system of the infested B. distachyon appeared smaller and the fibrous roots were shorter and less numerous. We found that a dose of 1 000 J2s of H. avenae was sufficient for nematode infestation. We showed that Koz-1 of B. distachyon could reproduce more cysts than TR2A line. Line Koz-1 also supported the complete life cycles of 5 CCN geographical populations belonging to the Ha1 or Ha3 pathotype group. Our results suggest that B. distachyon is a host for CCN.展开更多
基金financially supported by the National Natural Science Foundation of China(31972247)the Science and Technology Innovation Project of the Chinese Academy of Agricultural Sciences(ASTIP-2016-IPP-04)the Special Fund for Agro-scientific Research in the Public Interest,China(201503114)。
文摘Heterodera filipjevi continues to be a major threat to wheat production worldwide.Rapid detection and quantification of cyst nematodes are essential for more effective control against this nematode disease.In the present study,a TaqManminor groove binder(TaqMan-MGB)probe-based fluorescence quantitative real-time PCR(qPCR)was successfully developed and used for quantifying H.filipjevi from DNA extracts of soil.The primers and probe designed from the obtained RAPD-SCAR marker fragments of H.filipjevi showed high specificity to H.filipjevi using DNA from isolatesconfirmed species of 23 Heterodera spp.,1 Globodera spp.and 3 Pratylenchus spp.The qPCR assay is highly sensitive and provides improved H.filipjevi detection sensitivity of as low as 4^(-3) single second-stage juvenile(J2)DNAs,10^(-3) female DNAs,and 0.01μgμL^(-1) genomic DNAs.A standard curve relating to the threshold cycle and log values of nematode numbers was generated and validated from artificially infested soils and was used to quantify H.filipjevi in naturally infested field soils.There was a high correlation between the H.filipjevi numbers estimated from 32 naturally infested field soils by both conventional methods and the numbers quantified using the qPCR assay.qPCR potentially provides a useful platform for the efficient detection and quantification of H.filipjevi directly from field soils and to quantify this species directly from DNA extracts of field soils.
基金financially supported by the National Natural Science Foundation of China (31171827)the National Key Basic Research Program of China (973 Program,2013CB127502)
文摘The cereal cyst nematode（CCN, Heteroder aavenae） causes serious yield loss on cereal crops, especially wheat, worldwide. Daxing population in Beijing City and Huangyuan population in Qinghai Province, China, are two CCN populations. In this study, the CCN pathotypes of Daxing and Huangyuan populations were characterized by tests on 23 standard ＂International Test Assortment＂ with the local species Wenmai 19 as the susceptible control. Tested materials were grouped by three nematode populations＇ virulence on resistant genes（Rha1, Rha2, Rha3, Cre1） and nonresistant genes, varieties and lines. Both Daxing and Huangyuan populations were avirulent to Ortolan（Ha1）. Barley cvs. Ortolan, Siri, Morocco, Bajo Aragon 1-1, and Martin 403-2 were all resistant to both populations. Cultivars Herta, Harlan 43 and wheat Iskamish-K-2-light were all susceptible to Huangyuan population, all of them, however, were resistant to Daxing population. The other five oats were all resistant to the two tested CCN populations. Except Iskamisch K-2-light, all the other wheat cultivars（Capa, Loros×Koga, AUS 10894, and Psathias） were susceptible to Daxing population. Because the pathotypes of the two tested CCN populations in Beijing and Qinghai were not identical to any of the 13 pathotypes previously characterized by the test assortment, we classified Daxing and Huangyuan populations as the new pathotypes, named Ha91.
基金financially supported by the National Natural Sciences Foundation of China(31571988)the Special Fund for Agro-scientific Research in the Public Interest,China(201503114)
文摘The cereal cyst nematodes(Heterodera avenae,Heterodera filipjevi,Heterodera latipons)are considered to be one of the most important plant parasitic nematodes attacking most cereals and can cause significant crop losses(Sikora 1988).In China,H.filipjevi(Madzhidov 1981)Stelter,1984,was first reported from Henan province(peng et al.2010)and a few years later in Anhui province and Xinjiang Uygur Autonomous Region(peng et al.2016,2018).In December 2017,a survey for cereal
基金funded by the National Key Basic Research Program of China (2013CB127501)the Special Fund for Agro-scientific Research in the Public Interest in China (201503114 and 200903040)the National Key Research and Development Program of China (SQ2017ZY060063-01)
文摘Cereal cyst nematode（Heterodera avenae, CCN） distributes worldwide and has caused severe damage to cereal crops, and a model host will greatly aid in the study of this nematode. In this research, we assessed the sensitivity of 25 inbred lines of Brachypodium distachyon to H. avenae from Beijing, China. All lines of B. distachyon were infested by secondstage juveniles（J2s） of H. avenae from Daxing District of Beijing population, but only 13 inbred lines reproduced 0.2–3 cysts/plant, showing resistance. The entire root system of the infested B. distachyon appeared smaller and the fibrous roots were shorter and less numerous. We found that a dose of 1 000 J2s of H. avenae was sufficient for nematode infestation. We showed that Koz-1 of B. distachyon could reproduce more cysts than TR2A line. Line Koz-1 also supported the complete life cycles of 5 CCN geographical populations belonging to the Ha1 or Ha3 pathotype group. Our results suggest that B. distachyon is a host for CCN.
