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复合RT-PCR方法同步检测百合X病毒、百合无症病毒及百合斑驳病毒 总被引:6,自引:0,他引:6
建立了特异性检测百合X病毒(LVX)、百合无症病毒(LSV)及百合斑驳病毒(LMoV)的单一、双重及三重PCR体系并对各体系的灵敏度进行了研究。结果表明,单一PCR体系可检测的LVX最低浓度在1×10-7μg/mL,LSV最低浓度在1×10-8μg/mL,LMoV最低浓度在1pg/mL;双重PCR体系可明显检测的LVX、LSV最低浓度在1pg/mL,LSV、LMoV最低浓度在1ng/mL,LVX、LMoV最低浓度在1pg/mL;三重PCR体系可明显检测的LVX、LSV和LMoV的最低浓度为1ng/mL。 相似文献
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单抗I-ELISA和TAS-ELISA检测百合无症病毒的研究 总被引:1,自引:0,他引:1
以抗百合无症病毒(Lily symptom less virus,LSV)的单克隆抗体为核心,建立了间接ELISA (I-ELISA)和三抗体夹心ELISA (TAS-ELISA)的检测方法。I-ELISA检测体系中,病叶汁液和单克隆抗体腹水的工作浓度分别为1:20和1:6 000,对病叶汁液的检测灵敏度达到了1:2 560,可检测到提纯病毒绝对量为1.35 ng。TAS-ELISA检测体系中捕获抗体和单克隆抗体腹水的工作浓度分别为1:200和1:6 000,检测病叶汁液的灵敏度达到了1:5 120,对于提纯病毒可检测到0.68 ng。使用美国Agdia公司双抗体夹心ELISA (DAS-ELISA)检测试剂盒对病叶汁液的检测灵敏度为1:2 560,提纯病毒检出量1.35 ng。用3种ELISA方法检测了采自浙江省丽水市的46个田间样品,I-ELISA、TAS-ELISA和DAS-ELISA测出的阳性样品数分别为19、21和18个,阳性率为41%、46%和39%。灵敏度检测和田间样品检测结果显示,TAS-ELISA的灵敏度高于DAS-ELISA和I-ELISA。相同样品I-ELISA所测出的OD405值和P/N值普遍高于DAS-ELISA,表明LSV单抗比多抗具有更强的特异性和更高的灵敏度。 相似文献
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干旱胁迫对3种苹果砧木叶片光合、叶绿体超微结构和抗氧化系统的影响 总被引:1,自引:0,他引:1
以新疆野苹果(Malus sieversii Roem.(XJ))、垂丝海棠(Malus halliana Koehne(CS))和山定子(Malus baccata Borkh.(SDZ))3种一年生苹果砧木实生苗为材料,采用盆栽控水的方法设置正常供水(75%~80% RWC)和干旱胁迫(45%~50% RWC)2种水分处理,研究干旱胁迫下叶片光合特性、叶绿体超微结构、丙二醛(MDA)含量、超氧阴离子(O—[KG-1][JX*3]·[JX-*3]2)产生速率以及抗氧化酶活性的变化规律,并利用主成分分析(PCA)对3种砧木进行抗旱性综合评价。结果表明:干旱胁迫抑制了3种苹果砧木的光合作用,但干旱胁迫下CS的净光合速率(Pn)、气孔导度(Gs)、蒸腾速率(Tr)、PSII最大光能转化效率(Fv/Fm)、潜在光化学活性(Fv/Fo)及光化学猝灭系数(qP)的降幅均显著小于其他两种砧木,胞间CO2浓度(Ci)和非光化学猝灭系数(NPQ)的增幅显著高于其他两种砧木;干旱胁迫下,3种砧木超微结构受到不同程度的伤害,其中CS叶片的超微结构损伤较小,能较好地保持细胞结构的完整性;干旱胁迫下3种砧木的SOD和CAT活性先升高后降低,POD活性逐渐增加至21 d趋于稳定,MDA含量和O—[KG-1][JX*3]·[JX-*3]2产生速率持续升高。PCA结果显示:2个主成分的方差贡献率达到98.502%,干旱胁迫下CS的综合得分最高。因此,干旱胁迫下,垂丝海棠能保持叶绿体结构的完整性,激活抗氧化酶系统,清除氧化产物而保持较高的光合能力。 相似文献
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Y. B. Zhang Y. J. Wang Z. K. Xie R. Y. Wang G. Yang Z. H. Guo Y. Qiu 《Plant pathology》2019,68(2):261-268
Two viruses that frequently occur in many Lilium species are Lily mottle virus (LMoV) and Cucumber mosaic virus (CMV), which usually co-infect lilies causing severe disease symptoms. Recent reports have revealed that the viral coat protein (CP) affects chloroplast ultrastructure and symptom development. This study used western blot analysis to confirm that in leaves infected by mixed virus infections of LMoV and CMV, CPs of both viruses were accumulated in lily chloroplasts. Immunogold labelling further demonstrated that both the LMoV CP and CMV CP were localized in the stroma and the thylakoid membranes of the chloroplasts. In addition, it was found that CPs of both viruses were rapidly transported into isolated, intact chloroplasts (in vitro), and their transport efficiencies were positively related to CP concentrations. The lowest transmembrane concentration of CMV CP decreased from 38 μg mL−1 recorded in the single CMV CP import system to 10 μg mL−1 in the mixed import system of LMoV CP and CMV CP. CPs of both viruses exhibited species selection in their transmembrane transport into chloroplasts. This is the first report that the CPs from two viruses (LMoV and CMV) are simultaneously present in lily chloroplasts. Accumulation of high levels of LMoV CP and CMV CP inside the chloroplast appears to contribute to a synergistic interaction inducing the development of mosaic symptoms. 相似文献
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The coat protein sequences were characterized of Lily symptomless virus (LSV) isolates infecting Lilium longiflorum , Lilium tigrinum , Hymenocalis littoralis (spider lily) and Asiatic and Oriental hybrid lilies in India. The Indian isolates showed 78–96% homology with each other. With LSV isolates from elsewhere in the world, the Indian isolates showed 83–98% homology. The LSV-L ( L. longiflorum ) and LSV-A (Asiatic hybrid) isolates had unique stretches in the middle portion of the protein not found in other LSV isolates, even the Indian ones. The LSV gene sequence from the spider lily isolate (LSV-S) was reported for the first time outside the Liliaceae. LSV-S was 84–96% similar to the other Indian isolates at the protein level. The isolate infecting tiger lily (LSV-T) was found to be different from the characterized isolates from elsewhere in the world (78–84% homology at the protein level). At the same time, LSV-T showed much variability in the C-terminal of the protein. A stretch of 41 amino acids in the C-terminal was unique to this isolate. LSV-T is proposed as a distinct isolate of LSV infecting L. tigrinum indigenous to India. 相似文献
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硅对黄瓜霜霉病抑制效果和抗性相关酶活性的影响 总被引:3,自引:1,他引:3
在营养液中加入不同浓度硅且接种黄瓜霜霉病菌后,通过调查其病情指数和检测黄瓜叶片内硅元素含量及过氧化物酶(guaiacol-peroxidase,POD)、多酚氧化酶(polyphenol oxidase,PPO)、苯丙氨酸解氨酶(phenylalanine ammonia-lysae,PAL)、β-1,3葡聚糖酶(β-1,3-glucanase)、超氧化物歧化酶(superoxidedismutase,SOD)5种抗霜霉病相关酶活性,探讨硅抑制黄瓜霜霉病的生理生化机制。结果表明,营养液中硅浓度为100mg/L的处理,黄瓜霜霉病病情指数为21.3,防治效果达到62.8%;营养液中硅浓度与黄瓜叶片内硅元素含量呈正相关,200mg/L处理叶片内硅元素含量最高,且7天后达到2.98mg/g;加硅处理接种黄瓜霜霉病菌后,黄瓜叶片抗病相关酶活性变化明显且差异达显著水平,其中硅浓度为100~200mg/L时上述5种酶活性最高。 相似文献
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ALA对NaCl胁迫下不同品种番茄植株光合作用、保护酶活性及果实产量的影响 总被引:11,自引:0,他引:11
以耐盐性不同的番茄品种金鹏超冠(耐盐性较强)和中杂9号(耐盐性较弱)为材料,研究了叶面喷施5-氨基乙酰丙酸(ALA)对150 mmol/L,250 mmol/L NaCl胁迫下番茄光合作用、保护酶系统及产量的影响。结果表明,ALA能使NaCl胁迫下番茄叶片的叶绿素含量、净光合速率(Pn)、蒸腾速率(Tr)、气孔导度(Gs)升高,胞间二氧化碳浓度(Ci)降低;ALA处理显著提高了高盐胁迫下的SOD、POD和CAT活性,且降低了丙二醛含量,尤其是中杂9号更为显著,ALA处理可能是通过促进保护酶系统活性,保护细胞的光合性能,促进光合作用,从而促进番茄的生长发育,提高产量,缓解盐胁迫对番茄的抑制作用。 相似文献
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侵染百合的黄瓜花叶病毒亚组Ⅱ分离物cp基因克隆和序列分析 总被引:1,自引:0,他引:1
从云南大理的东方型百合上得到黄瓜花叶病毒分离物(CMV-DL), ELISA检测初步确定为CMV亚组Ⅱ分离物, 设计并合成CMV亚组Ⅱ的特异引物, RT-PCR扩增得到1条约800 nt的特异片段, 经克隆及序列测定, 该片段长828 nt, 包含的外壳蛋白(CP)基因由657 nt组成。将该分离物的cp基因与其它14个CMV分离物进行同源性比较, 在核苷酸水平上与CMV亚组I和亚组Ⅱ的同源性分别为76.8%~78.1%和98.6%~99.2%;在氨基酸水平上与CMV亚组I和亚组Ⅱ的同源性分别为82.0%~84.3%和95.9%~100.0%。结果表明CMV-DL为CMV亚组Ⅱ成员。 相似文献
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J. C. M. Beijersbergen Carola Th. C. Van Der Hulst 《European journal of plant pathology / European Foundation for Plant Pathology》1980,86(6):277-283
Lily symptomless virus (LSV) was readily detected with the enzyme-linked immunosorbent assay (ELISA) in leaves ofLilium Mid-Century hybrids cvs Enchantment and Destiny andLilium speciosum cv. Brabander. In bulbs of cv. Enchantment LSV could be detected without any additional treatment of the extracts, but for reliable results in cv. Destiny it was necessary to pre-incubate the bulb extracts with cellulase or hemicellulase.Samenvatting In de bladeren van secundair geïnfecteerde planten van de leliecultivars Enchantment, Destiny en Brabander kon LSV met ELISA gemakkelijk worden aangetoond. In bladextracten van Enchantment kon nog ca. 100 ng LSV/ml extract worden teruggevonden. Bij toepassing van de standaard-methode van extraheren (2 g weefsel malen in 4 ml extractiebuffer) kon ook in de bolschubben van Enchantment LSV worden aangetoond; bij Destiny gelukte dat aanzienlijk minder goed. Wijzigingen in de extractie-procedure of een speciale behandeling van het standaard-extract (dialyseren, verhitten tot 50°C etc.) leidde niet tot een betere aantoonbaarheid van het virus met ELISA. Enzymen als hemicellulase, cellulase en pectinase, toegevoegd aan het standaard-extract van bolschubben van Destiny vóór de toetsing (incubatie gedurende 18 h bij kamertemperatuur), verbeteren de aantoonbaarheid van het LSV aanzienlijk. 相似文献