• <tr id="yyy80"></tr>
  • <sup id="yyy80"></sup>
  • <tfoot id="yyy80"><noscript id="yyy80"></noscript></tfoot>
  • 99热精品在线国产_美女午夜性视频免费_国产精品国产高清国产av_av欧美777_自拍偷自拍亚洲精品老妇_亚洲熟女精品中文字幕_www日本黄色视频网_国产精品野战在线观看 ?

    Host status of Brachypodium distachyon to the cereal cyst nematode

    2018-02-05 07:10:49CHENChanglongLIUShusenLIUQianNIUJunhaiLIUPeiZHAOJianlongLIUZhiyongLIHongjieJIANHeng
    Journal of Integrative Agriculture 2018年2期

    CHEN Chang-long, LIU Shu-sen, LIU Qian NIU Jun-hai, LIU Pei ZHAO Jian-long LIU Zhi-yong,LI Hong-jie, JIAN Heng

    1 Department of Plant Pathology, China Agricultural University, Beijing 100193, P.R.China

    2 National Key Facility for Crop Gene Resources and Genetic Improvement (NFCRI), Institute of Crop Sciences, Chinese Academy of Agricultural Sciences, Beijing 100081, P.R.China

    3 Plant Protection Institute, Hebei Academy of Agricultural and Forestry Sciences/IPM Center of Hebei Province/Key Laboratory of IPM on Crops in Northern Region of North China, Ministry of Agriculture, Baoding 07100, P.R.China

    4 Tropical Crops Genetic Resources Institute, Chinese Academy of Tropical Agricultural Sciences, Danzhou 571737, P.R.China

    5 Department of Plant Genetics & Breeding, China Agricultural University, Beijing 100193, P.R.China

    6 Institute of Genetics and Developmental Biology, Chinese Academy of Sciences, Beijing 100101, P.R.China

    1. Introduction

    Cereal cyst nematode (Heterodera avenae, CCN), one of the most important plant parasitic nematodes (PPNs),has caused severe damage to cereal crops worldwide.H.avenaeis one of the most important pathogens of wheat(Triticum aestivumL.), which causes substantial yield losses ranging from 30–100% (Bonfilet al. 2004; Nicolet al. 2007; Penget al. 2009). It occurs in 16 provinces in China at high level of prevalence (Penget al. 2015).The development of molecular management strategies will undoubtedly be promoted if we know more about the mechanism of plant-H.avenaeinteraction.Arabidopsis thalianahas been reported as a model host in the study of plant-nematode interactions (Sijmonset al. 1991). This model host facilitates new findings, such as the linkage between cell cycle modifications and the differentiation of syncytia and giant cells, interactions between nematode effectors and plant targets, the role of auxin in feeding-site differentiation and transcriptomics (Joneset al. 2012).Unfortunately, relatively few nematode species can complete their life cycles onArabidopsiswith the exception ofH.avenae(Joneset al. 2012).

    As wheat has complex genetics and a lower efficacy for transformation, which limits study of plant-H.avenaeinteraction, an alternative host would be exceptionally useful.Brachypodium distachyon, a new monocot model plant system first proposed by Draperet al. (2001), was taken into consideration. Similar to wheat,B.distachyonbelongs to the Pooideae subfamily of the Poaceae family.The simple growth requirements ofB.distachyon, its small stature (approximately 30 cm at maturity), its short generation time (8–10 weeks), its self-fertility, and its small,fully sequenced diploid genome (approximately 272 Mbp for the Bd21 diploid accession) represent all the desirable features of a powerful plant model (Peraldiet al. 2014).B.distachyonis being developed as a model for grasses.This initiative is comparable to the development ofA.thalianaas a model for dicotyledonous plants. For example, efficient transformation protocols (P?curaret al.2008; Vainet al. 2008; Vogel and Hill 2008), germplasm collections (Vogelet al. 2006; Filizet al. 2009; Vogelet al.2009), genetic markers (Vogelet al. 2009), mutant collections(http://brachypodium.pw.usda.gov, http://www.brachytag.org), microarrays, and databases (http://www.brachybase.org, http://www.phytozome.net, http://www.modelcrop.org, http://mips.helmholtz-muenchen.de/plant/index.jsp)(IBI 2010) have been developed. These tools make it easy to conduct genetic and molecular experiments onB.distachyon. Furthermore, this species has been reported as a host of a number of pathogens, includingMagnaporthe grisea(Draperet al. 2001; Routledgeet al. 2004),Puccinia striiformis(Draperet al. 2001),P.brachypodii(Barbieriet al. 2011),Fusariumspecies (F.graminearumandF.culmorum) (Peraldiet al. 2011), arbuscular mycorrhizal fungi (Honget al. 2012), andBarley stripe mosaic virus(Cuiet al. 2012), making it a promising pathosystem model for studying plant-pathogen interactions.

    In this study, we assessed the host status of 25 lines ofB.distachyontoH.avenaefor their potential as an alternative host.

    2. Materials and methods

    2.1. Nematodes and inoculation

    The 5 different geographical populations ofH.avenaewere sampled from fields in the Daxing District of Beijing City(DX population, Ha3 pathotype group) (Su 2012); Baoding,Hebei Province (BD population, Ha1 pathotype group) (Liet al. 2014); Luannan, Hebei Province (LN population,Ha1 pathotype group) (Liet al. 2014); Xingyang, Henan Province (XY population, Ha3 pathotype group) (Yuanet al.2010); and Xuzhou, Jiangsu Province (XZ population, Ha1 pathotype group) (Liang 2014). They were identified by PCR amplification and sequencing of the internal transcribed spacer region (ITS).

    Infective second-stage juveniles (J2s) of CCN were obtained by hatching cysts at 15°C after at least 2 months of incubation at 4°C. The J2 water suspension was inoculated into 2 holes in the soil per plant seedling. The holes were 2 cm deep across and closed to each plant seedling.

    2.2. Plant materials and growth conditions

    A total of 25 inbred lines ofB.distachyonand their origins are listed in Table 1. Wheat seeds (Triticum aestivum cv.Aikang 58) were purchased from Henan Bainong Seed Co., Ltd., Henan, China. Seeds ofB.distachyonwere embedded in Petri dishes on damp filter paper for 1 day at 25°C followed by incubation for 7 days at 4°C. The wheat seeds were then surface-sterilised for 5 min in 3% NaClO,raised with water and soaked in water for 1 day at room temperature before they were placed in Petri dishes on damp filter paper for 1 day at 25°C. The seeds were then planted in 5.5 cm×5.5 cm×5.5 cm pots filled with sterilised 75% sand mixed with 25% sandy-loam soil. Plants were grown in an artificial environment at 22–25°C with 16 h light/8 h dark photoperiod. When theB.distachyonseedlings were at the 3–4 euphylla stage or the wheat seedlings were approximately 10 cm high, they were inoculated with J2s ofH.avenae, and the inoculated plants were grown at 16°C for 10 days or longer forH.avenaeinfestation. Then plants were grown at 20–25°C to allow the development ofH.avenaefor 3 mon or more until all of the cysts formed and fell into the soil.

    2.3. Screening of the B. distachyon lines

    A total of 600 J2s ofH.avenaefrom DX per plant were inoculated twice (300 J2s each time) into 25 inbred lines ofB.distachyonat an interval of 10 days, respectively.The nematodes invading into each plant root system were stained with acid fuchsin and counted at 15 and 30 days post the first inoculation (dpi) with 3 plant replicates for each inbred line at each sampling time. The cysts/plant were extracted from the soil and counted for each line with 5–9 plant replicates at 5 mon post the first inoculation (mpi),except for line Adi-3 with 4 plant replicates. Wheat was used as a control with 3 plant replicates. To further evaluate the sensitivity of Koz-1 and TR2A to nematode, 600 J2s/plant were inoculated into each line in a single inoculation with 9 plant replicates. The total number of cysts/plant was counted at 3 mpi.

    Table 1 Average number of cereal cyst nematode (CCN) on Brachypodium distachyon lines and wheat observed at various times1)

    2.4. Phenotype of B. distachyon infested with CCN

    A total of 0, 1 000 or 2 000 J2s of the DX population ofH.avenaeper plant were inoculated into Bd21-3 and wheat,respectively. Plant height (10 plant replicates) and root length (5 plant replicates) were assessed at 5 dpi, and plant height was assessed again at 45 dpi (5 plant replicates). The typical root symptoms were photographed. The nematodes inside the roots were stained and counted at 5 dpi (4–5 plant replicates for each nematode dosage).

    2.5. Infestation of different geographical populations of H. avenae to B. distachyon

    A total of 600 J2s from each of the 5 geographical populations ofH.avenaewere inoculated into line Koz-1,respectively. There were 9 plant replicates for each geographical population. The average numbers of cyst/plant were compared among 5 geographical populations after 3 mon inoculation. Wheat was used as a control with 6–9 plant replicates per geographical population.

    2.6. Staining and microscopy

    The roots were washed with tap water and stained using the sodium hypochlorite-acid fuchsin method (Bybdet al.1983). The nematodes in or on the roots were observed and photographed under a stereoscopic microscope (OLYMPUS SZ-61, New York, America).

    2.7. Statistical analysis

    The data were analyzed by a one-way ANOVA or an independent-samplest-test performed in SPSS 13.0.

    3. Results

    3.1. Half of the lines of B. distachyon produced cysts of H. avenae

    Each of the 25 lines ofB.distachyonwas successfully infested by CCN J2s of DX population, while only 13 lines reproduced cysts (0.2–3 cysts/plant) (Table 1), showing resistance. Representative figures of different life stages of CCN on Bd21-3 are shown in Fig. 1, including J2 (Fig. 1-A),third-stage juvenile (J3) (Fig. 1-B), fourth-stage juvenile (J4)(Fig. 1-C), a mature female cyst full of eggs (Fig. 1-D), and J2s hatched from newly formed cyst (Fig. 1-E). Of these 13 lines ofB.distachyon, young nematode females were observed on the root surface of lines Gaz-4, Bd21-3, Koz-4,Gaz-8, Koz-1, and BdTR2A at 30 dpi (0.3–1 cysts/plant)(Table 1), indicating that CCN developed faster on these lines than on other lines.

    3.2. Phenotypes of B. distachyon infested by CCN

    Fig. 1 Representative figures of the life cycle of Heterodera avenae on line Bd21-3 of Brachypodium distachyon. The arrows indicate second-stage juvenile (J2) (A), third-stage juvenile (J3) (B), fourth-stage juvenile (J4) (C), and a mature female cyst full of eggs (D) and J2s hatched from the cyst (E).Scale bar=200 μm.

    Fig. 2 Phenotypes of line Bd21-3 of Brachypodium distachyon and wheat infected by Heterodera avenae. A total of 0, 1 000 or 2 000 second-stage juveniles (J2s) of H. avenae were used to inoculate each plant, and representative root symptom photographs of Bd21-3 (A) and wheat cultivar Aikang 58 (B) were taken at 5 days post the first inoculation (dpi). Consistent with wheat (B),the fibrous roots of Bd21-3 (A) infected by H. avenae also became shorter and fewer compared to the uninfected roots, and the severity of symptom was enhanced with the increase of nematodes inoculated (1 000 J2s vs. 2 000 J2s).

    We qualitatively observed the phenotype of the aerial and underground parts ofB.distachyoninfested by CCN using Bd21-3 line as a representative. Similar to wheat (Fig. 2-B),the entire root system of Bd21-3 infested by CCN appeared to be smaller, and the fibrous roots were also shorter and fewer compared to those of non-inoculated roots at 5 dpi(Fig. 2-A). In addition, the symptom was more obvious in the plants inoculated with 2 000 J2s than those inoculated with 1 000 J2s (Fig. 2-A, B). However, the root length of Bd21-3 (15.0, 14.2, and 12.0 cm/plant for 0, 1 000, and 2 000 J2s, respectively) was not affected by nematode infestation at 5 dpi, whereas that of the wheat cultivar Aikang 58 was affected (19.4, 15.5, and 15.9 cm/plant for 0, 1 000, and 2 000 J2s, respectively) (Fig. 3-A). Plant heights of Bd21-3 and wheat were not affected by CCN infestation evaluating at 5 dpi (5.6–5.9 and 17.5–18.9 cm/plant for Bd21-3 and wheat,respectively) (P>0.05, Fig. 3-B) and 45 dpi (21.3–24.4 and 33.0–35.4 cm/plant for Bd21-3 and wheat, respectively)(P>0.05, Fig. 3-C).

    3.3. Comparison of nematode dosages for B. distachyon

    We used 1 000 and 2 000 J2s/plant to assess the upper limit of the dosage of the DX population of CCN onB.distachyon.There were no differences between the average numbers of nematodes invading into the roots at 5 dpi either for Bd21-3(43.5 and 33.3 J2s/plant for 1 000 and 2 000 J2s dosage,respectively) or for wheat (400.8 and 467.0 J2s/plant for 1 000 and 2 000 J2s dosage, respectively) (P>0.05, Fig. 4).Accordingly, 1 000 J2s/plant would represent the upper limit of the inoculation dosage of the nematode.

    3.4. The most suitable B. distachyon line for CCN infestation

    For further study, we would like to choose one line that is relatively more suitable for CCN parasitism and reproduction. According to our initial screening results,Koz-1 can reproduce 3 cysts/plant on average and up to 6 cysts individually. In both cases, these values were the maxima for the evaluated plant lines (Table 1). To confirm the initial result, we conducted another infection assay using Koz-1 and TR2A, the latter plant line can also produce 1.2±1.3 (0–3) cysts/plant in our initial screening (Table 1).The results indicated that Koz-1 reproduced more cysts(4.6±2.2 cysts/plant) than did TR2A (1.7±1.4 cysts/plant)(P<0.01, Fig. 5), indicating that Koz-1 is more suitable as a host, at least for the DX population of CCN.

    3.5. Five different geographical populations of CCN can complete their life cycle on line Koz-1

    Line Koz-1 can support the life cycle of each of the 5 different geographical populations ofH.avenae(belonging to the Ha1 or Ha3 pathotype group) with only moderate variations(3.6–10.3 cysts/plant) (Fig. 6), suggesting the general compatibility of Koz-1. TheH.avenaepopulation from XZ developed the greatest number of cysts in both Koz-1 (10.3 cysts/plant) and wheat (248.7 cysts/plant) (P<0.05) (Fig. 6).

    4. Discussion

    Fig. 3 Comparison of the average root length at 5 days post the first inoculation (dpi) (n=5) (A), the average plant height at 5 dpi(n=10) (B) or at 45 dpi (n=5) (C) among 0, 1 000, and 2 000 second-stage juveniles (J2s) for line Bd21-3 of Brachypodium distachyon and for wheat cultivar Aikang 58. Each column shows the mean and standard deviation. Statistical analyses were conducted for Bd21-3 and wheat independently. Columns labelled with the same letter for Bd21-3 or Aikang 58 do not differ significantly (P>0.05).

    Currently,B.distachyonis being developed as a model for grasses similar toA.thalianafor dicotyledonous plants.Previous results showedB.distachyonis a promising pathosystem model for the study of plant-fungi or virus interactions (Draperet al. 2001; Routledgeet al. 2004;Barbieriet al. 2011; Peraldiet al. 2011; Honget al. 2012; Cuiet al. 2012). In this research, we evaluated the sensitivity ofB.distachyontoH.avenae. Our results showed that certain lines ofB.distachyoncould support the complete life cycle ofH.avenae. Similar to wheat, the fibrous roots of the infestedB.distachyonby nematode were obviously fewer and shorter. This indicatesB.distachyonis a host of CCN though not very susceptible. It was reported that Bd21-3 was not a host for CCN, as nematodes penetrated roots but failed to develop to the later developmental stages or form cysts (Konget al. 2016). While in this research,Bd21-3 formed 0.2±0.4 (0–1) cysts/plant, which was not consistent with previous report. These two contrary results might be due to different CCN populations inoculated, that is, we inoculated DX CCN while Konget al. (2016) used population they maintained in a greenhouse by themselves.

    In the evaluation assay on wheat resistance toH.avenae,500–800 J2s/plant were always inoculated (Liet al. 2012).In theB.distachyoninoculation, this study showed the upper limit of theH.avenaenematode dosage was 1 000 J2s/plant. AsB.distachyonhas smaller root systems than wheat does, we recommend to innoculateB.distachyon600 J2s/plant for saving nematode. There are more than 13 pathotypes belonging to 3 pathotype groups (Ha1, Ha2 and Ha3) forH.avenae(Yuanet al. 2010). We assessed susceptibility ofB.distachyonto 5 different geographical populations ofH.avenaewhich belong to 2 pathotype groups.B.distachyon(line Koz-1) can support the complete life cycles of 5 different geographical populations ofH.avenae. These results indicate the general compatibility of Koz-1 for the infection and development ofH.avenae.However, different geographical populations ofH.avenaedeveloped different numbers of cysts onB.distachyon. In addition, on wheat, Ha1 pathotype group (XZ, BD and LN)ofH.avenaedeveloped more cysts than Ha3 pathotype group (DX and XY), while there isn’t the same case onB.distachyon(line Koz-1).

    Fig. 4 Average number of nematode in roots of line Bd21-3 of Brachypodium distachyon and wheat cultivar Aikang 58 inoculated with 1 000 and 2 000 second-stage juveniles(J2s)/plant at 5 days post the first inoculation (dpi) (n=4–5).Each column shows the mean and standard deviation.Statistical analyses were conducted for Bd21-3 and Aikang 58 independently. Columns labelled with the same letter for Bd21-3 or wheat did not differ significantly (P>0.05).

    Fig. 5 Comparison of the average cyst number of DX population(from the Daxing District of Beijing City, China) of cereal cyst nematode (CCN) reproduced on Koz-1 and TR2A. Each column shows the average cyst number of DX population of CCN/plant and the standard deviation of 9 plant replicates of each line inoculated with 600 J2s/plant. Columns labelled with different letters indicate significant difference between the average cyst numbers of DX population of CCN obtained from Koz-1 and TR2A (P<0.01).

    However, this study also raises several questions.Firstly, the cysts that reproduced onB.distachyonroots were not as abundant as those on wheat. The cysts from Koz-1 ofB.distachyoninfected by the 5 different geographical populations ofH.avenaeexamined ranged from 3.6 to 10.3/plant, while those on wheat ranged from 121.8 to 248.7/plant. One reason might be the smaller root system ofB.distachyoncompared to that of wheat.Another reason might be the tested plant lines are not as susceptible as wheat to the 5 geographical populations ofH.avenaeexamined. As there are large numbers of collections ofB.distachyon(Brkljacicet al. 2011), additional screening experiments should be performed to identify whether there are more susceptible lines toH.avenaein the future. Another confusion in our preliminary screening research was that the average number of nematodes per plant of line BdTR2A (up to 40.7±17.2) was the greatest when observed at 30 dpi, but only few cysts were ultimately formed (1.2±1.3). The reason for the incapacity of these later-stage nematodes to complete their life cycles is not clear, but it is likely associated with the late induction of plant resistance. As Joneset al. (2012) have stated,plant resistance to nematodes can occur not only at early infection stage but also after the successful establishment of a feeding site. For example, during the interactions between resistant cowpea containing theR-geneRkandMeloidogyne incognita, normal nematode feeding occurred,and nematodes were able to develop to later juvenile stages,but giant cell deterioration was observed, and the female nematodes showed arrested development and deterioration and never reached maturity (Daset al. 2008). The same may be the case for the interaction of DX population of CCN and BdTR2A, and observations of syncytium are needed in the future to confirm this hypothesis.

    Fig. 6 Average cyst numbers of Koz-1 or wheat inoculated with 600 second-stage juveniles (J2s) of different geographical populations of cereal cyst nematode (CCN) per plant. Each column shows the mean cyst numbers in 6–9 plant replicates and the standard deviation. Statistical analyses were conducted for Koz-1 and wheat cultivar Aikang 58 independently, and columns labelled with the same letter do not differ significantly(P>0.05). XZ, Xuzhou of Jiangsu Province, China; BD, Baoding of Hebei Province, China; LN, Luannan of Hebei Province,China; DX, Daxing District of Beijing City, China; XY, Xingyang of Henan Province, China.

    5. Conclusion

    Our results suggested that the monocotyledonous model plantB.distachyoncan be infected by the cereal cyst nematodeH.avenaeand a certain lines ofB.distachyoncould support the whole life cycle ofH.avenae. The infection symptoms of the whole root ofB.distachyonwere nearly as same as that of wheat. Furthermore, we found that Koz-1 was the most suitable line for CCN infestation in our study,and Koz-1 can support different geographical populations of CCN which indicates the universality of the line. More lines ofB.distachyonshould be assessed for their susceptibility to CCN in the future and the mechanisms of susceptibility variation can be studied further.

    Acknowledgements

    We are grateful to Profs. Chen Shulong and Li Xiuhua (Plant Protection Institute, Hebei Academy of Agricultural and Forestry Sciences, Baoding, China), Li Honglian (Henan Agricultural University, Zhengzhou, China), and Li Hongmei(Nanjing Agricultural University, Nanjing, China) for helping with collections or for providing different geographical populations ofHeterodera avenae. This study was 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) and the National Key Research and Development Program of China (SQ2017ZY060063-01).

    Barbieri M, Marcel T C, Niks R E. 2011. Host status of false brome grass to the leaf rust fungusPuccinia brachypodiiand the stripe rust fungusP.striiformis.Plant Disease, 95,1339–1345.

    Bonfil D J, Dolgin B, Mufradi I, Asido S. 2004. Bioassay to forecast cereal cyst nematode damage to wheat in fields.Precision Agriculture, 5, 329–344.

    Brkljacic J, Grotewold E, Scholl R, Mockler T, Garvin D F, Vain P, Caicedo A L. 2011.Brachypodiumas a model for the grasses: Today and the future.Plant Physiology, 157, 3–13.

    Bybd D W, Kirkpatrick Jr T, Barker K R. 1983. An improved technique for clearing and staining plant tissues for detection of nematodes.Journal of Nematology, 15, 142–143.

    Cui Y, Lee M Y, Huo N, Bragg J, Yan L, Yuan C, Li C, Holditch S J, Xie J, Luo M C, Li D, Yu J, Martin J, Schackwitz W, Gu Y Q, Vogel J P, Jackson A O, Liu Z, Garvin D F. 2012. Fine mapping of theBsr1barley stripe mosaic virus resistance gene in the model grassBrachypodium distachyon.PLoS ONE, 7, e38333.

    Das S, DeMason D A, Ehlers J D, Close T J, Roberts P A.2008. Histological characterization of root-knot nematode resistance in cowpea and its relation to reactive oxygen species modulation.Journal of Experimental Botany, 59,1305–1313.

    Draper J, Mur L A, Jenkins G, Ghosh-Biswas G C, Bablak P, Hasterok R, Routledge A P. 2001.Brachypodium distachyon. A new model system for functional genomics in grasses.Plant Physiology, 127, 1539–1555.

    Filiz E, Ozdemir B S, Budak F, Vogel J P, Tuna M, Budak H.2009. Molecular, morphological, and cytological analysis of diverseBrachypodium distachyoninbred lines.Genome,52, 876–890.

    Hong J J, Park Y S, Bravo A, Bhattarai K K, Daniels D A,Harrison M J. 2012. Diversity of morphology and function in arbuscular mycorrhizal symbioses inBrachypodium distachyon.Planta, 236, 851–865.

    IBI (The International Brachypodium Initiative). 2010. Genome sequencing and analysis of the model grassBrachypodium distachyon.Nature, 463, 763–768.

    Jones J, Gheysen G, Fenoll C. 2012.Genomics and Molecular Genetics of Plant-Nematode Interactions. Springer, London& New York.

    Kong L A, Wu D Q, Cui J K, Huang W K, Peng H, Peng D L.2016. Testing and modelling the potential of three diploid plants in Poaceae as a new pathosystem to investigate the interactions between cereal hosts and cereal cyst nematode(Heterodera avenae).Plant Pathology, 65, 682–688.

    Li X H, Gu S J, Wang W L, Chen S L. 2012. Effect of different factors on number ofHeterodera avenaecysts produced on the wheat.Journal of Agricultural University of Hebei,35, 56–61. (in Chinese)

    Li X H, Ma J, Gao B, Wang R Y, Chen S L. 2014. Characterization of the pathotypes of the cereal cyst nematode,Heterodera avenae, in Hebei Province.Plant Protection, 40, 127–131.(in Chinese)

    Liang X D. 2014. Pathotype identification ofHeterodera avenae,resistant evaluation of wheat materials and determing of damage threshold. MSc thesis, Nanjing Agricultural University, China. (in Chinese)

    Nicol J M, Elek?io?lu I H, Bolat N, Rivoal R. 2007. The global importance of the cereal cyst nematode (Heteroderaspp.)on wheat and international approaches to its control.Communications in Agricultural and Applied Biological Sciences, 72, 677–686.

    P?curar D I, Thordal-Christensen H, Nielsen K K, Lenk I. 2008.A high-throughputAgrobacterium-mediated transformation system for the grass model speciesBrachypodium distachyonL.Transgenic Research, 17, 965–975.

    Peng D L, Nicol J M, Li H M, Hou S Y, Li H X, Chen S L, Ma P, Li H L, Riley I T. 2009. Current knowledge of cereal cyst nematode (Heterodera avenae) on wheat in China. In: Riley I T, Nicol J M, Dababat A A, eds.,Cereal Cyst Nematodes:Status,Research and Outlook.Proceedings of the First Workshop of the International Cereal Cyst Nematode Initiative. 21–23 Oct., 2009. International Maize and Wheat Improvement Centre (CIMMYT), Antalya. pp. 29–34.

    Peng D L, Peng H, Huang W K. 2015. Occurrence, distribution and integrated management of the cereal cyst nematodes(Heterodera avenae&H.filipjevi) in China. In: Dababat A A, Muminjanov H, Smiley R W, eds.,Nematodes of Small Grain Cereals: Current Status and Research. FAO, Ankara,Turkey. pp. 17–24.

    Peraldi A, Beccari G, Steed A, Nicholson P. 2011.Brachypodium distachyon: A new pathosystem to study Fusarium head blight and otherFusariumdiseases of wheat.BMC Plant Biology, 11, 100.

    Peraldi A, Griffe L L, Burt C, McGrann G R D, Nicholson P. 2014.Brachypodium distachyonexhibits compatible interactions withOculimaculaspp. andRamularia collo-cygni, providing the first pathosystem model to study eyespot and ramularia leaf spot diseases.Plant Pathology, 63, 554–562.

    Routledge A P, Shelley G, Smith J V, Talbot N J, Draper J, Mur L A. 2004.Magnaporthe griseainteractions with the model grassBrachypodium distachyonclosely resemble those with rice (Oryza sativa).Molecular Plant Pathology, 5, 253–265.

    Sijmons P C, Grundler F M W, Mende N, Burrows P R, Wyss U. 1991.Arabidopsis thalianaas a new model host for plant-parasitic nematodes.The Plant Journal, 1, 245–254.

    Su Z. 2012. Investigation on the major specie, pathotype of cereal cyst nematode in Beijing and evaluation of wheat cultivars resistance toHeterodera avenae.MSc thesis, Jilin Agricultural University, China. (in Chinese)

    Vain P, Worland B, Thole V, McKenzie N, Alves S C,Opanowicz M, Fish L J, Bevan M W, Snape J W. 2008.Agrobacterium-mediated transformation of the temperate grassBrachypodium distachyon(genotype Bd21) for T-DNA insertional mutagenesis.Plant Biotechnology Journal, 6,236–245.

    Vogel J, Hill T. 2008. High-efficiencyAgrobacterium-mediated transformation ofBrachypodium distachyoninbred line Bd21-3.Plant Cell Reports, 27, 471–478.

    Vogel J P, Garvin D F, Leong O M, Hayden D M. 2006.Agrobacterium-mediated transformation and inbred line development in the model grassBrachypodium distachyon.Plant Cell,Tissue and Organ Culture, 84, 199–211.

    Vogel J P, Tuna M, Budak H, Huo N, Gu Y Q, Steinwand M A. 2009. Development of SSR markers and analysis of diversity in Turkish populations ofBrachypodium distachyon.BMC Plant Biology, 9, 88.

    Yuan H, Sun J, Yang W, Xing X, Wang Z, Riley I T, Li H.2010. New pathotypes ofHeterodera avenae(cereal cyst nematode) from winter wheat in Zhengzhou, Henan, China.Australasian Plant Pathology, 39, 107–111.

    日本猛色少妇xxxxx猛交久久| 久久久久视频综合| 又大又爽又粗| 老汉色av国产亚洲站长工具| 久久国产精品男人的天堂亚洲| 亚洲欧美激情在线| 一区二区三区四区激情视频| 大片电影免费在线观看免费| 男人操女人黄网站| 成年美女黄网站色视频大全免费| 99国产精品免费福利视频| 国产乱人偷精品视频| 亚洲欧美一区二区三区国产| 高清不卡的av网站| 亚洲欧洲日产国产| 在线观看免费午夜福利视频| 久久精品国产亚洲av高清一级| 无限看片的www在线观看| 亚洲伊人久久精品综合| 伊人亚洲综合成人网| 黄色一级大片看看| 成年动漫av网址| 亚洲av在线观看美女高潮| 一级黄片播放器| 中文字幕精品免费在线观看视频| 黄色怎么调成土黄色| 桃花免费在线播放| 如日韩欧美国产精品一区二区三区| 国产 一区精品| 51午夜福利影视在线观看| 欧美激情极品国产一区二区三区| 久久久欧美国产精品| 国产亚洲av片在线观看秒播厂| 国产片特级美女逼逼视频| 国产免费视频播放在线视频| 国产视频首页在线观看| 国产成人欧美在线观看 | 欧美人与性动交α欧美软件| 黄色怎么调成土黄色| 国产精品 欧美亚洲| 啦啦啦在线观看免费高清www| 又大又黄又爽视频免费| 亚洲激情五月婷婷啪啪| 中文字幕人妻丝袜制服| 人人妻人人澡人人爽人人夜夜| 午夜福利,免费看| avwww免费| 亚洲成人免费av在线播放| 中文字幕亚洲精品专区| 一区二区三区激情视频| 一二三四在线观看免费中文在| 午夜福利影视在线免费观看| 黄色毛片三级朝国网站| 啦啦啦中文免费视频观看日本| 高清不卡的av网站| 看免费av毛片| 亚洲av电影在线观看一区二区三区| 精品久久蜜臀av无| av福利片在线| 少妇被粗大猛烈的视频| 成人手机av| 亚洲av中文av极速乱| 国产伦人伦偷精品视频| 免费高清在线观看日韩| www.av在线官网国产| 1024香蕉在线观看| 免费少妇av软件| 99久久人妻综合| 久久久久久久大尺度免费视频| 黄色视频在线播放观看不卡| 国语对白做爰xxxⅹ性视频网站| 美女福利国产在线| 国产精品女同一区二区软件| 国产精品 欧美亚洲| 国产男人的电影天堂91| 精品一区二区免费观看| 最黄视频免费看| 亚洲天堂av无毛| 91国产中文字幕| 亚洲av国产av综合av卡| 天天躁夜夜躁狠狠久久av| 国产成人精品久久二区二区91 | 国产熟女午夜一区二区三区| 十八禁人妻一区二区| 亚洲人成电影观看| 亚洲av国产av综合av卡| 一级,二级,三级黄色视频| 中文天堂在线官网| 欧美在线一区亚洲| 国产成人精品久久久久久| 一级毛片黄色毛片免费观看视频| 精品酒店卫生间| 不卡视频在线观看欧美| 捣出白浆h1v1| 国产在线免费精品| 黄色视频不卡| 中文字幕人妻丝袜一区二区 | 国产精品免费大片| 黑人猛操日本美女一级片| 久久久精品94久久精品| xxx大片免费视频| av网站在线播放免费| 午夜久久久在线观看| 国产激情久久老熟女| 免费久久久久久久精品成人欧美视频| 久久久久国产一级毛片高清牌| 一本色道久久久久久精品综合| 十分钟在线观看高清视频www| 别揉我奶头~嗯~啊~动态视频 | 亚洲综合色网址| 亚洲av成人精品一二三区| 久久久久精品久久久久真实原创| 又大又黄又爽视频免费| 日韩精品有码人妻一区| 亚洲一卡2卡3卡4卡5卡精品中文| 操出白浆在线播放| 日韩伦理黄色片| 久久久久精品久久久久真实原创| 国产极品粉嫩免费观看在线| 午夜av观看不卡| 免费人妻精品一区二区三区视频| 亚洲av国产av综合av卡| 中文欧美无线码| 久久性视频一级片| 欧美人与性动交α欧美精品济南到| 日本av免费视频播放| 制服诱惑二区| 少妇人妻 视频| 精品一区二区免费观看| 一本—道久久a久久精品蜜桃钙片| 色综合欧美亚洲国产小说| 午夜激情久久久久久久| av一本久久久久| 精品第一国产精品| 一区二区三区四区激情视频| 一区二区三区四区激情视频| 亚洲色图综合在线观看| 91精品国产国语对白视频| 大片免费播放器 马上看| 一级毛片我不卡| 欧美人与性动交α欧美软件| 老司机影院成人| 老司机在亚洲福利影院| 亚洲欧美一区二区三区国产| 在线观看www视频免费| 丝袜在线中文字幕| 日韩制服丝袜自拍偷拍| 免费少妇av软件| 国产午夜精品一二区理论片| 99re6热这里在线精品视频| 亚洲成色77777| 一级毛片黄色毛片免费观看视频| 人成视频在线观看免费观看| 亚洲一码二码三码区别大吗| 久久精品国产亚洲av高清一级| 1024视频免费在线观看| 男人操女人黄网站| 国产精品秋霞免费鲁丝片| 最新在线观看一区二区三区 | 中文字幕另类日韩欧美亚洲嫩草| 久久人妻熟女aⅴ| 日韩人妻精品一区2区三区| 只有这里有精品99| 又大又黄又爽视频免费| 国产男人的电影天堂91| 国产视频首页在线观看| 亚洲精品av麻豆狂野| 99九九在线精品视频| 丝袜脚勾引网站| 美女视频免费永久观看网站| 亚洲精品一区蜜桃| 无遮挡黄片免费观看| 日韩欧美一区视频在线观看| 中文字幕人妻丝袜一区二区 | 黄色怎么调成土黄色| 精品国产国语对白av| 久久久久精品性色| 欧美 日韩 精品 国产| 涩涩av久久男人的天堂| 国产av一区二区精品久久| 一本一本久久a久久精品综合妖精| 五月开心婷婷网| 欧美激情高清一区二区三区 | 丰满少妇做爰视频| 老司机影院成人| 精品国产超薄肉色丝袜足j| 国产日韩欧美在线精品| 亚洲视频免费观看视频| 亚洲国产日韩一区二区| 黑丝袜美女国产一区| 最近中文字幕2019免费版| 51午夜福利影视在线观看| 国产精品国产三级专区第一集| 国产欧美日韩综合在线一区二区| 国产男人的电影天堂91| 国产精品女同一区二区软件| 亚洲成人手机| www日本在线高清视频| 国产乱来视频区| 欧美亚洲日本最大视频资源| 91成人精品电影| av视频免费观看在线观看| 国产成人精品福利久久| 1024视频免费在线观看| av片东京热男人的天堂| 男女国产视频网站| 久久97久久精品| 国产老妇伦熟女老妇高清| 飞空精品影院首页| 久久精品熟女亚洲av麻豆精品| 亚洲国产精品999| 午夜福利在线免费观看网站| 观看av在线不卡| 精品一区二区三区四区五区乱码 | 精品国产一区二区三区久久久樱花| 国产黄色免费在线视频| 精品国产一区二区久久| 亚洲精品国产色婷婷电影| 欧美日韩国产mv在线观看视频| 国产黄色视频一区二区在线观看| 在线亚洲精品国产二区图片欧美| 无遮挡黄片免费观看| 欧美黑人精品巨大| 久久天堂一区二区三区四区| 五月开心婷婷网| 人成视频在线观看免费观看| 精品国产乱码久久久久久小说| 18禁裸乳无遮挡动漫免费视频| 国产精品亚洲av一区麻豆 | tube8黄色片| kizo精华| 99国产综合亚洲精品| 亚洲人成电影观看| xxx大片免费视频| 亚洲,一卡二卡三卡| 免费观看性生交大片5| 不卡av一区二区三区| 亚洲成人一二三区av| 精品亚洲成国产av| 在线精品无人区一区二区三| 人体艺术视频欧美日本| 亚洲一区二区三区欧美精品| 在线观看免费日韩欧美大片| 国产97色在线日韩免费| 黄片播放在线免费| 美女福利国产在线| 人人妻人人添人人爽欧美一区卜| 卡戴珊不雅视频在线播放| 91精品国产国语对白视频| 国产精品一区二区精品视频观看| 日日啪夜夜爽| 成年人免费黄色播放视频| 亚洲精品日韩在线中文字幕| 国产一区亚洲一区在线观看| 啦啦啦中文免费视频观看日本| a 毛片基地| 丁香六月天网| 美女高潮到喷水免费观看| 国产黄色免费在线视频| 日本午夜av视频| 久久久精品94久久精品| 亚洲国产欧美网| 欧美亚洲日本最大视频资源| 日韩精品免费视频一区二区三区| 宅男免费午夜| 中文天堂在线官网| 日韩精品有码人妻一区| 精品少妇久久久久久888优播| 老司机亚洲免费影院| 波多野结衣av一区二区av| 久久鲁丝午夜福利片| 一二三四在线观看免费中文在| 久久久久久久大尺度免费视频| 久久久欧美国产精品| 最近最新中文字幕大全免费视频 | 国产精品国产三级专区第一集| 操美女的视频在线观看| 中国国产av一级| 丝袜脚勾引网站| 国产又爽黄色视频| 女人久久www免费人成看片| 亚洲精品久久久久久婷婷小说| 成年女人毛片免费观看观看9 | 自线自在国产av| 亚洲久久久国产精品| 天天躁夜夜躁狠狠久久av| 成人国产av品久久久| 一区在线观看完整版| 国产一区二区 视频在线| 欧美精品亚洲一区二区| 波多野结衣av一区二区av| 亚洲av电影在线进入| 日本av免费视频播放| 国产激情久久老熟女| av不卡在线播放| 国产欧美日韩一区二区三区在线| 日韩,欧美,国产一区二区三区| 日本vs欧美在线观看视频| 一二三四在线观看免费中文在| 伦理电影大哥的女人| 菩萨蛮人人尽说江南好唐韦庄| 久久午夜综合久久蜜桃| 黄色视频在线播放观看不卡| 日韩制服丝袜自拍偷拍| 捣出白浆h1v1| av在线观看视频网站免费| 美女扒开内裤让男人捅视频| 最近手机中文字幕大全| 免费黄网站久久成人精品| 高清视频免费观看一区二区| 亚洲一区二区三区欧美精品| 欧美日韩亚洲国产一区二区在线观看 | 亚洲精品久久午夜乱码| 国产精品国产三级国产专区5o| 国产精品久久久人人做人人爽| 中文字幕色久视频| 一区二区三区四区激情视频| 免费看av在线观看网站| 免费在线观看完整版高清| 免费人妻精品一区二区三区视频| 男女边吃奶边做爰视频| 国产在视频线精品| 18禁观看日本| 看免费成人av毛片| 日本wwww免费看| 丰满少妇做爰视频| 国产亚洲一区二区精品| 国产精品香港三级国产av潘金莲 | 19禁男女啪啪无遮挡网站| 国产黄频视频在线观看| 中文字幕最新亚洲高清| 免费黄色在线免费观看| 人妻 亚洲 视频| 国产伦人伦偷精品视频| 国产精品秋霞免费鲁丝片| 女人爽到高潮嗷嗷叫在线视频| 亚洲av成人不卡在线观看播放网 | 日韩成人av中文字幕在线观看| 久久人人爽人人片av| 久久毛片免费看一区二区三区| 18在线观看网站| 黄片播放在线免费| 亚洲精品中文字幕在线视频| 欧美黑人欧美精品刺激| 午夜日本视频在线| av一本久久久久| 一边摸一边做爽爽视频免费| 两个人免费观看高清视频| 国产成人精品无人区| 欧美国产精品一级二级三级| 免费黄色在线免费观看| 亚洲精华国产精华液的使用体验| 久久久亚洲精品成人影院| 伦理电影免费视频| 一区二区三区激情视频| 午夜福利,免费看| 欧美精品一区二区免费开放| 人人澡人人妻人| 18禁动态无遮挡网站| videosex国产| 最新在线观看一区二区三区 | 一级毛片 在线播放| 欧美 亚洲 国产 日韩一| 午夜影院在线不卡| 一级片免费观看大全| 欧美国产精品一级二级三级| 日韩一区二区三区影片| 宅男免费午夜| 最黄视频免费看| 午夜91福利影院| 在线观看人妻少妇| 在线天堂最新版资源| 美女扒开内裤让男人捅视频| 国产亚洲av高清不卡| 伦理电影大哥的女人| e午夜精品久久久久久久| 老司机影院成人| 久热这里只有精品99| 这个男人来自地球电影免费观看 | 欧美变态另类bdsm刘玥| 91aial.com中文字幕在线观看| 国产成人免费无遮挡视频| 精品亚洲成国产av| av网站免费在线观看视频| 青春草亚洲视频在线观看| 晚上一个人看的免费电影| 亚洲av电影在线进入| 各种免费的搞黄视频| 国产黄色视频一区二区在线观看| 国产精品二区激情视频| 最黄视频免费看| 亚洲欧美一区二区三区久久| 天天影视国产精品| 电影成人av| 久久综合国产亚洲精品| 九九爱精品视频在线观看| 黑丝袜美女国产一区| 国产精品人妻久久久影院| 午夜激情av网站| 亚洲,欧美,日韩| 亚洲欧美成人综合另类久久久| 一本久久精品| 国产淫语在线视频| 亚洲人成电影观看| 久久97久久精品| 久久精品国产a三级三级三级| 99国产综合亚洲精品| 国产精品亚洲av一区麻豆 | 欧美激情高清一区二区三区 | 少妇精品久久久久久久| 大话2 男鬼变身卡| av天堂久久9| 飞空精品影院首页| 香蕉丝袜av| 久久午夜综合久久蜜桃| 久久久久久人妻| 久久精品国产亚洲av高清一级| 黑人欧美特级aaaaaa片| 久久精品国产a三级三级三级| 国产成人精品久久二区二区91 | 国产 精品1| 亚洲国产日韩一区二区| 亚洲成国产人片在线观看| 精品少妇一区二区三区视频日本电影 | 一区二区日韩欧美中文字幕| 国产精品无大码| 99九九在线精品视频| 一区二区日韩欧美中文字幕| av福利片在线| av女优亚洲男人天堂| 日本黄色日本黄色录像| 国产成人欧美在线观看 | 欧美日韩一级在线毛片| 超碰成人久久| 人人妻人人爽人人添夜夜欢视频| 观看av在线不卡| 久久久久精品性色| 19禁男女啪啪无遮挡网站| 777米奇影视久久| 亚洲av综合色区一区| 久久人妻熟女aⅴ| 精品第一国产精品| 国产一区二区 视频在线| 精品少妇一区二区三区视频日本电影 | 80岁老熟妇乱子伦牲交| 99香蕉大伊视频| 亚洲国产精品成人久久小说| 老汉色av国产亚洲站长工具| 爱豆传媒免费全集在线观看| 亚洲av综合色区一区| 天堂中文最新版在线下载| 天美传媒精品一区二区| 国产成人系列免费观看| 国产一区二区三区av在线| 午夜福利影视在线免费观看| 亚洲成av片中文字幕在线观看| 中文精品一卡2卡3卡4更新| 婷婷成人精品国产| 亚洲七黄色美女视频| 黑人欧美特级aaaaaa片| 日本91视频免费播放| 青草久久国产| 国产精品嫩草影院av在线观看| 19禁男女啪啪无遮挡网站| 大片电影免费在线观看免费| 悠悠久久av| 麻豆av在线久日| 日韩欧美精品免费久久| 欧美老熟妇乱子伦牲交| 十分钟在线观看高清视频www| 久久久精品区二区三区| 制服丝袜香蕉在线| 看免费av毛片| 亚洲精品久久成人aⅴ小说| 精品国产一区二区久久| 天堂俺去俺来也www色官网| 黄频高清免费视频| 日本av手机在线免费观看| 美女中出高潮动态图| 亚洲美女视频黄频| 麻豆精品久久久久久蜜桃| 亚洲成色77777| 国产精品一区二区在线不卡| 十八禁高潮呻吟视频| 制服诱惑二区| 国产精品99久久99久久久不卡 | 黄片播放在线免费| 久久狼人影院| 免费不卡黄色视频| 国产精品久久久久久精品古装| 纵有疾风起免费观看全集完整版| 国精品久久久久久国模美| 午夜福利乱码中文字幕| 99久久精品国产亚洲精品| 蜜桃国产av成人99| 日韩人妻精品一区2区三区| 精品少妇内射三级| kizo精华| 一本—道久久a久久精品蜜桃钙片| 国产一级毛片在线| 男女边摸边吃奶| 中文字幕精品免费在线观看视频| 捣出白浆h1v1| 亚洲欧洲精品一区二区精品久久久 | 国产精品久久久久久久久免| 老鸭窝网址在线观看| 欧美乱码精品一区二区三区| 精品少妇黑人巨大在线播放| 欧美国产精品一级二级三级| 波多野结衣av一区二区av| 最黄视频免费看| 秋霞在线观看毛片| 国产免费福利视频在线观看| 色播在线永久视频| 亚洲国产日韩一区二区| 久久午夜综合久久蜜桃| 成年动漫av网址| 久久久久视频综合| 亚洲美女搞黄在线观看| 国产精品偷伦视频观看了| 亚洲欧美中文字幕日韩二区| 性高湖久久久久久久久免费观看| 综合色丁香网| 熟女av电影| www.熟女人妻精品国产| 欧美精品高潮呻吟av久久| 亚洲美女视频黄频| 国产精品久久久久久久久免| 国产黄色免费在线视频| 免费日韩欧美在线观看| 人体艺术视频欧美日本| 亚洲,一卡二卡三卡| 免费在线观看完整版高清| 久久久久精品国产欧美久久久 | 国产女主播在线喷水免费视频网站| 天天躁日日躁夜夜躁夜夜| av有码第一页| 日本爱情动作片www.在线观看| 久久亚洲国产成人精品v| 国产精品三级大全| 国产日韩欧美在线精品| 国产97色在线日韩免费| 黑人巨大精品欧美一区二区蜜桃| 免费观看a级毛片全部| 亚洲三区欧美一区| 麻豆av在线久日| 国产精品99久久99久久久不卡 | 99久久综合免费| 看免费成人av毛片| 校园人妻丝袜中文字幕| 免费在线观看完整版高清| 激情五月婷婷亚洲| avwww免费| 又粗又硬又长又爽又黄的视频| 久久久国产欧美日韩av| 日韩一卡2卡3卡4卡2021年| 久久久久久人妻| 欧美日韩一级在线毛片| 国产精品久久久久久久久免| 午夜av观看不卡| 久久久精品区二区三区| 国产黄色免费在线视频| 9热在线视频观看99| 欧美精品人与动牲交sv欧美| av女优亚洲男人天堂| 高清黄色对白视频在线免费看| 我要看黄色一级片免费的| 亚洲成人av在线免费| 午夜日韩欧美国产| 久久久国产精品麻豆| 毛片一级片免费看久久久久| 性高湖久久久久久久久免费观看| 免费观看性生交大片5| 人人妻人人爽人人添夜夜欢视频| 精品一区在线观看国产| 赤兔流量卡办理| 欧美久久黑人一区二区| 欧美老熟妇乱子伦牲交| 久久午夜综合久久蜜桃| 亚洲视频免费观看视频| 久久ye,这里只有精品| kizo精华| 青青草视频在线视频观看| 人妻一区二区av| 久久久久久久久免费视频了| 人妻一区二区av| 亚洲情色 制服丝袜| 99re6热这里在线精品视频| 人人妻人人添人人爽欧美一区卜| 午夜福利,免费看| 91老司机精品| 日本黄色日本黄色录像| 久久97久久精品| 岛国毛片在线播放| 欧美另类一区| 国产乱人偷精品视频| 久久久精品国产亚洲av高清涩受| 久久久久久免费高清国产稀缺| 免费观看人在逋| 男男h啪啪无遮挡| 永久免费av网站大全| 丰满少妇做爰视频| 国产熟女欧美一区二区| 亚洲精品国产av成人精品| 久久人妻熟女aⅴ| av女优亚洲男人天堂| 久久免费观看电影| 韩国av在线不卡| 免费人妻精品一区二区三区视频| 一区二区三区激情视频| 香蕉国产在线看| 欧美 日韩 精品 国产| 一区二区av电影网| 涩涩av久久男人的天堂| 久久久久国产一级毛片高清牌|