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1.
【目的】明确柑桔全爪螨Panonychus citri(Mc Gregor)高致病力菌株芽枝状枝孢霉Cladosporium cladosporioides(CQBBW3)的生物学特性。【方法】通过单因素试验,测试了不同培养基、碳源、氮源、温度及pH对CQBBW3菌株生长和产孢的影响。【结果】CQBBW3菌株的LC50为4.66×108个孢子/m L,LT50为6.2353 d。最佳生长培养基是淀粉琼脂培养基,最适产孢培养基为察氏培养基;菌丝生长最适碳源和氮源为甘露醇和甘氨酸,产孢最适碳源和氮源为乳糖和赖氨酸;最适生长温度为25℃,最适产孢温度为15℃和30℃;最适生长pH值为6.0~8.0,最适产孢pH值7.0。【结论】CQBBW3菌株对柑桔全爪螨有较好的杀虫活性,喜好25℃、中性、富含有机营养的环境。本研究为菌株快速培养及其真菌杀虫制剂的开发与田间应用奠定基础。  相似文献   

2.
生防细菌T132的鉴定及其对采后柑橘炭疽病的抑制效果   总被引:2,自引:0,他引:2  
【目的】柑橘(Citri)是世界上重要的果树。由胶孢炭疽菌[Colletotrichum gloeosporioides(Penz.)]引起的柑橘炭疽病是柑橘生产的主要病害之一。为探索对采后柑橘炭疽病有效的生防措施,分离鉴定柑橘根围土壤中一株细菌T132,并研究其特性及生防效果。【方法】根据菌株T132的形态特征、生理生化特性以及16S rDNA序列对其进行鉴定;通过连续8次在人工培养基上传代培养,测定该菌株的遗传稳定性;采用柑橘果实刺伤挑战接种和拮抗菌液直接浸泡健康果实两种方法研究该菌株对柑橘炭疽病的抑菌防病效果;利用洋葱伯克霍尔德氏菌致病因子的特异性引物检测菌株T132是否为潜在的人类致病菌。【结果】菌株T132鉴定为越南伯克霍尔德氏菌(Burkholderia vietnamiensis)。连续8次在人工培养基上传代培养,菌株T132抑制胶孢炭疽病菌生长的能力没有发生明显改变。菌株T132对胶孢炭疽菌C.gloeosporioides引起的柑橘炭疽病有明显的防治作用,刺伤接种的防效为88.2%,自然发病的防效为54.9%。未检测到该拮抗菌株有人体致病相关的洋葱伯克霍尔德氏菌致病因子(BCESM)毒力基因。【结论】首次报道对柑橘采后炭疽病具有生防效果、对人类相对安全的越南伯克霍尔德氏菌生防菌株。  相似文献   

3.
【目的】筛选柑橘木虱Diaphorina citri Kuwayama应对球孢白僵菌侵染的免疫应答及其网络调控基因,以进一步探讨柑橘木虱对球孢白僵菌的免疫防御机制。【方法】采用Illumina高通量测序平台对感染球孢白僵菌24、48、72 h与健康的柑橘木虱转录组进行了测序,利用生物信息学软件对筛选到的差异表达基因进行了功能注释、分类以及参与的信号通路分析。【结果】组装得到138 313条不可延长的非冗余Unigene,其N50和N90分别为2 532 bp和413 bp,平均长度为1 191.26 bp。在CK vs.S24h、CK vs.S48h和CK vs.S72h 3个转录组里分别获得了971、1 671和752个显著差异表达基因(DEGs),GO富集分析表明分别有405、614和542个DEGs显著富集到56、83和60个GO terms中,KEGG pathway分析结果显示分别有98、333和247个DEGs显著富集到10、27和25条代谢通路里。【结论】差异表达基因主要富集在能量代谢、离子运输、转录和翻译调控、生殖和发育调控以及免疫防御反应等相关通路,大部分编码潜在的与免疫识别及调控相关的基因,并从中筛选到5个显著上调表达的免疫相关基因,为开展柑橘木虱免疫应答虫生真菌的研究奠定理论基础。下一步将进行免疫相关基因的q-PCR验证及表达量分析,研究其在柑橘木虱应对球孢白僵菌侵染过程中的作用。  相似文献   

4.
【目的】昆虫速激肽相关肽(Tachykinin-related peptides,TRPs)属于肽类神经递质,参与调控昆虫生长、发育、变态、繁殖等多种生命活动。本文旨在探究柑橘木虱Diaphorina citri速激肽相关肽(Dc-TRP)基因序列信息,进一步分析其在不同发育时期及成虫不同组织中的表达情况。【方法】本文克隆得到柑橘木虱速激肽相关肽全长序列,并根据核苷酸系列进行系列分析及结构预测。利用RT-q PCR对柑橘木虱不同发育时期及成虫不同组织部位Dc-TRP的表达情况进行分析。【结果】(1)Dc-TRP编码区全长600 bp,预测编码199个氨基酸,具有多个保守区域。(2)时空表达结果表明Dc-TRP在各发育时期和组织均有表达,其中各发育历期表达差异不明显,而组织中表达差异显著且头部表达最高。【结论】Dc-TRP在柑橘木虱不同发育时期表达无显著性差异,但在成虫不同组织中差异显著。以上实验结果具有一定的生物学意义,为进一步研究Dc-TRP的生理功能提供一定的理论基础。  相似文献   

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【目的】柑橘全爪螨Panonychus citri在中国是一种重要的柑橘害虫,叶面肥在橘园的应用很普遍。本研究是为了明确柑橘施用尿素和复合氨基酸2种叶面肥对这种害螨生长发育和繁殖及柑橘苗生长的影响。【方法】在室内分别用尿素(0.50%)和复合氨基酸(0.17%)2种叶面肥喷施盆栽沙糖橘Citrus reticulata Blanco cv. Shatangju苗,以喷施清水为对照,探究叶面施肥对柑橘全爪螨生命表参数[净 增殖率(R0)、平均代时(T)、内禀增长率(rm)、周限增长率(λ)和种群趋势指数(I)]及柑橘苗生长参数(叶长、宽和面积, 茎长, 株高, 新梢的长度和数量)和叶片养分(N, P和K)含量的影响。【结果】柑橘全爪螨未成熟螨态的发育历期没有受到叶面肥的影响,但施用0.50%尿素的柑橘苗上第2若螨的存活率(95.40%)显著高于施用清水的对照(78.26%)和喷施0.17%复合氨基酸的处理(75.61%),其雌螨的繁殖力(42.1/♀)也显著高于对照(33.1/♀)。复合氨基酸处理柑橘苗上的雌螨寿命(19.5 d)显著长于尿素处柑橘苗上的雌螨寿命(14.8 d)和对照(14.5 d),复合氨基酸处理柑橘苗上的雄螨寿命(17.6 d)也显著长于对照(13.1 d)。总体上,在尿素处理的柑橘苗上柑橘全爪螨的净增殖率(R0)(17.88)和种群趋势指数(I)(18.08)值最高,2个参数都显著高于对照(分别为10.08和11.17)。施用2种叶面肥显著促进了柑橘苗叶片生长(叶长、叶宽、叶面积),其N, P和K含量以及氮钾比(N/K)也显著增加。【结论】柑橘苗叶面喷施尿素和复合氨基酸都可促进柑橘苗生长,喷施尿素会导致柑橘全爪螨种群的显著增长,而喷施复合氨基酸没有导致柑橘全爪螨种群显著增长。因此,推荐使用复合氨基酸代替尿素作为柑橘的叶面肥施用。但是,喷施复合氨基酸可显著延长柑橘全爪螨成螨的寿命,所以在使用时还应该加强对其种群的监测。  相似文献   

6.
【目的】研究柑橘木虱Diaphorina citri Kuwayama在柚、酸橘、黄皮、九里香、砂糖橘5种代表性芸香科寄主植物上的发育、存活和繁殖情况,为柑橘木虱及黄龙病的可持续防控提供参考。【方法】利用种群生命表的方法,分析了柑橘木虱在5种不同寄主植物上的发育历期、存活率、成虫寿命、性比及产卵量等数据。【结果】柑橘木虱卵、1龄若虫以及整个若虫的发育历期受寄主植物的影响较为明显,在26℃条件下在柚子植物上柑橘木虱若虫的存活率最高(58.10%),在黄皮上最低(46.04%),两者差异显著。寄主植物影响柑橘木虱成虫寿命,在柚子上柑橘木虱成虫的寿命显著长于黄皮上的寿命。黄皮上的柑橘木虱的单雌产卵量(298粒/雌)显著低于其他4种寄主植物。柑橘木虱在九里香上的内禀增长率(rm)最高(0.133 7)、酸橘上最低(0.129 8);而净增值率(R0)在砂糖橘上最高(187.74)、黄皮上最低(145.27)。【结论】在5种寄主植物中,除黄龙病隐症寄主九里香之外,显症寄主中砂糖橘是柑橘木虱的最适寄主。  相似文献   

7.
【目的】比较宛氏拟青霉WS-11和球孢白僵菌QB-28对柑橘木虱成虫的致病力,为生防制剂的田间应用提供理论支持。【方法】采用孢子悬浮液浸没法比较研究了2种真菌对柑橘木虱的致病力,并利用时间-剂量-死亡率模型估计了2种真菌对柑橘木虱的致死剂量与致死时间。【结果】宛氏拟青霉WS-11在孢子悬浮液浓度为1×108 spores/m L时,累计死亡率达到90.67%,球孢白僵菌QB-28则达到97.33%。时间-剂量-死亡率模型中Hosmer-Lemeshow方法拟合异质性检验表明模型拟合良好,在接种后9 d,宛氏拟青霉WS-11和球孢白僵菌QB-28对柑橘木虱的LC50分别为7.57×10~6 spores/m L和8.39×105 spores/m L;当孢子悬浮液浓度为1×108 spores/m L时,2种真菌对柑橘木虱的LT50分别为2.50 d和1.93 d。【结论】宛氏拟青霉WS-11和球孢白僵菌QB-28对柑橘木虱均有较好的致病力,具有较好的生防潜质,其中球孢白僵菌对柑橘木虱的致病力高于宛氏拟青霉,致死效率更高。  相似文献   

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【背景】栽培基质的利用是广叶绣球菌(Sparassis latifolia)栽培中重要的生理过程,但栽培过程中基质代谢物的变化尚不清楚。【目的】通过不同生长阶段栽培基质中差异代谢物分析挖掘关键代谢物,为广叶绣球菌基质利用机理研究提供理论参考。【方法】利用UHPLC-MS/MS技术分析广叶绣球菌菌丝(Myc)、原基(Pri)和子实体(FB)生长阶段栽培基质中代谢产物的变化,通过不同数据库进行代谢物注释并进行KEGG通路富集分析。利用LC-MS/MS技术检测不同发育阶段绣球菌中植物类激素含量。【结果】三个不同栽培阶段基质中共鉴定出代谢产物1 360个。不同比较组(Pri vs. Myc、FB vs. Myc和FB vs. Pri)间共有的差异代谢产物179个,含量最高的50个代谢物主要包括氨基酸、脂质、吡喃酸、吡喃酮和植物类激素等物质。其中氨基酸含量在Myc、Pri和FB阶段基质中逐渐降低,而吡喃酸和吡喃酮类化合物含量逐渐升高。植物类激素中的赤霉素在Pri和FB阶段基质中含量较高,茉莉酸在Myc阶段基质中含量较高。对不同发育阶段绣球菌植物类激素进行检测,发现赤霉素GA7仅在原基中检测到,1...  相似文献   

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【背景】龋病危害着人类健康,变异链球菌(Streptococcusmutans)是公认的主要致龋菌。我们前期的研究从口腔中分离出一株枯草芽孢杆菌(Bacillussubtilis),其能抑制S.mutans的生长,但尚未明确其抑菌作用的组分。【目的】利用非靶向代谢组学技术检测分析B. subtilis zh78抑制S. mutans生长可能的小分子代谢物,以评估其在龋病防治研究及应用中的前景。【方法】取B. subtilis zh78经生长0 h、7 h、12 h、5 d时的菌液,利用牛津杯法将冷甲醇萃取后的代谢物对S. mutans进行抑菌实验;采用气相色谱-飞行时间质谱联用技术(GC-TOF-MS)对代谢物进行检测;利用主成分分析(Principalcomponentanalysis,PCA)、正交偏最小二乘判别分析(Orthogonalpartialleastsquaresdiscriminantanalysis,OPLS-DA)和皮尔森相关分析(Pearson correlationanalysis)等方法进行数据分析。【结果】B.subtiliszh78生长5d时的代谢产物对S.mutans具有明显抑菌作用,其代谢物中木糖醇、氨基酸类及有机酸类等36种物质与其对S.mutans的抑制作用显著相关。【结论】B. subtilis zh78可能产生木糖醇、氨基酸类及有机酸类等可能促进B. subtilis zh78抑制S. mutans生长的小分子代谢产物,具有一定的防龋应用研究前景。  相似文献   

10.
【目的】了解亚洲柑橘木虱Diaphorina citri Kuwayama与黄龙病菌Canditatus Liberibacter asiaticus的分子互作机制。【方法】利用转录组(RNA-Seq)方法分别对携带和未携带黄龙病菌的柑橘木虱成虫进行转录组测序,根据获得的转录信息分析木虱携带黄龙病菌后基因表达的差异。【结果】获得了1 502个差异表达的unigenes,在带菌木虱中上调和下调的基因分别有746和756个。共有1 099个unigenes比对上NCBI蛋白数据库并获得功能注释;852个被聚类到GO的三大功能中;931个被注释到KEGG的239个代谢通路中。根据差异表达基因的GO分析,木虱代谢过程和催化活性的相关基因明显上调。此外,筛选出53个免疫相关的unigenes中,28个与细胞免疫、25个与免疫信号路径相关,分别有64%和32%表达上调。【结论】黄龙病菌和媒介昆虫柑橘木虱存在互作关系,病菌入侵后可能通过代谢活动相关基因的上调和免疫相关基因的差异表达而影响木虱的代谢活动和免疫反应过程。  相似文献   

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目的甲型H1N1流感病毒A/California/7/2009与A/California/4/2009病毒序列比较同源性在99%以上,本实验旨在比较两株病毒感染BALB/c小鼠研究感染力强弱。方法分别将A/California/7/2009(CA7)与A/California/4/2009(CA4)两株病毒分别连续10倍稀释后,对4~6周龄雌性BALB/c小鼠经乙醚麻醉后进行滴鼻攻毒,每个稀释度接种10只实验小鼠,测定CA7 MLD50为101.24/0.05 mL,检测小鼠感染、致病的多项指标,观察期为14 d。结果相同TCID50的CA7和CA4病毒感染小鼠,CA4感染小鼠后14 d内死亡率为20%,而CA7感染小鼠后8 d内死亡率为100%。CA7 106TCID50感染的小鼠病理表现为重度弥漫性间质性肺炎,CA4 106TCID50感染的小鼠病理表现为中度-重度间质性肺炎。结论在相同条件下,CA7感染力明显强于CA4。  相似文献   

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SSeCKS/Gravin/AKAP12 (“SSeCKS”) encodes a cytoskeletal protein that regulates G1 → S progression by scaffolding cyclins, protein kinase C (PKC) and PKA. SSeCKS is down-regulated in many tumor types including prostate, and when re-expressed in MAT-LyLu (MLL) prostate cancer cells, SSeCKS selectively inhibits metastasis by suppressing neovascularization at distal sites, correlating with its ability to down-regulate proangiogenic genes including Vegfa. However, the forced re-expression of VEGF only rescues partial lung metastasis formation. Here, we show that SSeCKS potently inhibits chemotaxis and Matrigel invasion, motility parameters contributing to metastasis formation. SSeCKS suppressed serum-induced activation of the Raf/MEK/ERK pathway, resulting in down-regulation of matrix metalloproteinase-2 expression. In contrast, SSeCKS had no effect on serum-induced phosphorylation of the Src substrate, Shc, in agreement with our previous data that SSeCKS does not inhibit Src kinase activity in cells. Invasiveness and chemotaxis could be restored by the forced expression of constitutively active MEK1, MEK2, ERK1, or PKCα. SSeCKS suppressed phorbol ester-induced ERK1/2 activity only if it encoded its PKC binding domain (amino acids 553–900), suggesting that SSeCKS attenuates ERK activation through a direct scaffolding of conventional and/or novel PKC isozymes. Finally, control of MLL invasiveness by SSeCKS is influenced by the actin cytoskeleton: the ability of SSeCKS to inhibit podosome formation is unaffected by cytochalasin D or jasplakinolide, whereas its ability to inhibit MEK1/2 and ERK1/2 activation is nullified by jasplakinolide. Our findings suggest that SSeCKS suppresses metastatic motility by disengaging activated Src and then inhibiting the PKC-Raf/MEK/ERK pathways controlling matrix metalloproteinase-2 expression and podosome formation.  相似文献   

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Eleven recessive mutant loci define the class of cop / det / fus mutants of Arabidopsis. The cop / det / fus mutants mimic the phenotype of light-grown seedlings when grown in the dark. At least four cop / det / fus mutants carry mutations in subunits of the COP9 signalosome, a multiprotein complex paralogous to the 'lid' subcomplex of the 26S proteasome. COP1, another COP/DET/FUS protein, is itself not a subunit of the COP9 signalosome. In the dark, COP1 accumulates in the nucleus where it is required for the degradation of the HY5 protein, a positive regulator of photomorphogenesis. In the light, COP1 is excluded from the nucleus and the constitutively nuclear HY5 protein can accumulate. Nuclear accumulation of COP1 and degradation of HY5 are impaired in the cop / det / fus mutants that carry mutations in subunits of the COP9 signalosome. Although the cellular function of the COP/DET/FUS proteins is not yet well understood, taken together the current findings suggest that the COP/DET/FUS proteins repress photomorphogenesis in the dark by mediating specific protein degradation.  相似文献   

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16.
应用反向遗传学技术,选择冷适应、温度敏感、减毒的A/Ann Arbor/6/60 ca (H2N2)型流感病毒的6个内部基因为骨架,与A/California/07/2009株流感病毒2个抗原基因HA、NA分别克隆到polⅠ-polⅡ转录表达载体pAD3000中,构建8个转录表达载体重组质粒,共转染Vero细胞,获得重配A/California/07/2009ca株流感病毒.重配病毒的TCID50为7.5,病毒传4代后其血凝素(HA)滴度稳定在1∶256,半数感染剂量EID50为8,鸡胚传20代,经RT-PCR鉴定未发现重组病毒基因突变,电镜观察重配病毒符合流感病毒的主要特征;蔗糖纯化的病毒经肌肉注射(灭活)及滴鼻(减毒活病毒)两种途径免疫BALB/c小鼠,结果显示:滴鼻免疫和肌肉注射都可以产生较高效价的血凝抑制(HI)抗体,肌肉注射组产生的HI抗体略高(P = 0.044),但肌肉注射组检测不到高效价IgA抗体;滴鼻免疫组鼻冲洗液中可以检测到高效价的IgA抗体,同型病毒感染后,IL-1β、TNFα、IFN-α等前炎因子分泌较早,且高于肌肉注射组(P < 0.05),可见,喷鼻减毒疫苗比灭活全病毒疫苗能更好地激发黏膜免疫反应.通过对小鼠各个器官病毒载量的检测发现,4天后鼻腔、气管、脑、肺、脾脏没有病毒存在,证明减毒活疫苗株在小鼠上是安全的.以上数据可以初步断定,重组病毒有作疫苗候选株的可能,而且喷鼻疫苗具有降低免疫剂量、同时激活体内体液免疫和细胞免疫的功能.  相似文献   

17.
We performed a detailed investigation of N-glycan structures on BM-40 purified from different sources including human bone, human platelets, mouse Engelbreth-Holm-Swarm (EHS) tumor, and human BM-40 recombinantly expressed in 293 and osteosarcoma cells. These preparations were digested with endoglycosidases and N-glycans were further characterized by sequential exoglycosidase digestion and high-performance liquid chromatography (HPLC) analyses. Bone BM-40 carries high-mannose structures as well as biantennary complex type N-glycans, whereas the protein from platelets and 293 cells has exclusively bi- and triantennary complex type structures. BM-40 derived from the EHS tumor carries biantennary complex type and additional hybrid structures. Using the osteosarcoma-derived MHH-ES1 cell line we successfully expressed a recombinant BM-40 that bears at least in part the bone-specific high-mannose N-glycosylation in addition to complex type and hybrid structures. Using chromatography on Concanavalin-A Sepharose, we further purified a fraction enriched in high-mannose structures. This array of differentially glycosylated BM-40 proteins was assayed by surface plasmon resonance measurements to investigate the binding to collagen I. BM-40 carrying high-mannose structures binds collagen I with higher affinity, suggesting that differentially glycosylated forms may have different functional roles in vivo.  相似文献   

18.
Viscumin of mistletoe (Viscum album L.) has a concentration-dependent activity profile unique to plant AB-toxins. It starts with lectin-dependent mitogenicity and then covers toxicity and cell agglutination, associated with shifts in the monomer/dimer equilibrium. Each lectin subunit harbors two sections for ligand contact. In the dimer, the B-chain sites in subdomain 2 gamma (designated as the Tyr-sites) appear fully accessible, whereas Trp-sites in subdomain 1 alpha are close to the dimer interface. It is unclear whether both types of sites operate similarly in binding glycoligands in solution. By systematically covering a broad range of lactose/lectin ratio in isothermal titration calorimetry, we obtained evidence for two sites showing dissimilar binding affinity. Intriguingly, the site with higher affinity was only partially occupied. To assign the observed properties to the Trp/Tyr-sites, we next performed chemically induced dynamic nuclear polarization measurements of Trp and Tyr accessibility. A Tyr signal, but not distinct Trp peaks, was recorded when testing the dimer. Lactose-quenchable Trp peaks became visible on the destabilization of the dimer by citraconylation, intimating Trp involvement in ligand contact in the monomer. Fittingly, Tyr acetylation but not mild Trp oxidation reduced the dimer hemagglutination activity and the extent of binding to asialofetuin-Sepharose 4B. Altogether, the results attribute lectin activity in the dimer primarily to Tyr-sites. Full access to Trp-sites is gained on dimer dissociation. Thus, the monomer/dimer equilibrium of viscumin regulates the operativity of these sites. Their structural divergence affords the possibility for differences in ligand selection when comparing monomers (Tyr- and Trp-sites) with dimers (primarily Tyr-sites).  相似文献   

19.
To create core/shell/shell quantum dots (QDs) with high stability against a harmful chemical environment, CdTe/CdS QDs were coated with a ZnO shell in an aqueous solution. An interfaced CdS layer sandwiched between a CdTe core and ZnO shell provided relaxation of the strain at the core/shell interface since lattice parameters of CdS are intermediate between those of CdTe and ZnO. The photoluminescence (PL) peak wavelength of the core/shell/shell QDs was shifted from 569 to 615 nm by adjusting the size of CdTe cores and thickness of CdS and ZnO shells, along with the highest PL quantum yield of the core/shell/shell QDs reaching 80%, which implies promising applications in the field of biomedical labeling. Due to the decrease of surface defects, it was observed that PL lifetimes significantly increased at room temperature as follows: 29.6 34.2, and 47.5 ns for CdTe (537 nm), CdTe/CdS (555 nm) and CdTe/CdS/ZnO (581 nm) QDs, respectively. Copyright © 2012 John Wiley & Sons, Ltd.  相似文献   

20.
概述了生物信息学中的一些研究方向及分析方法,介绍了用于大规模DNA测序的分析软件系统——Phred/Phrap/Consed。通过利用Phred/Phrap/Consed等各种分析软件,时基因组学、蛋白质组学和基因芯片研究中巨量原始实验数据进行分析、处理,使之成为具有明确生物学意义的生物信息。  相似文献   

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