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1.
为明确热激蛋白HSP10在沙葱萤叶甲Galeruca daurica生长发育过程中的作用,采用RTPCR及RACE技术克隆沙葱萤叶甲Hsp10基因cDNA的全长序列,采用生物信息学方法分析其序列特征,并利用qPCR技术对该基因在沙葱萤叶甲不同发育阶段、成虫羽化后不同时期以及不同温度下的表达谱进行分析。结果表明,克隆获得1条新的沙葱萤叶甲Hsp10基因,命名为GdHsp10a,GenBank登录号为MG460308,cDNA序列全长为526 bp,开放阅读框为333 bp,编码蛋白含110个氨基酸,预测分子量为11.97 kD,等电点为9.74;无信号肽及跨膜结构。同源比对及系统进化分析表明,GdHSP10a与光肩星天牛Anoplophora glabripennis HSP10亲缘关系最近,氨基酸序列一致性为53.15%。qPCR结果表明,GdHsp10a在沙葱萤叶甲各发育阶段均有表达,其中在卵期和成虫期的表达量显著高于幼虫期、预蛹期及蛹期;成虫羽化后不同时期表达量差异显著,25 d时表达量最高,其次为100、10和7 d时的表达量;温度对GdHsp10a表达量有显著影响,30℃下表达量最高,35℃下表达量次之,15、20、25及40℃下表达量最低且无显著差异。表明GdHsp10a可能在沙葱萤叶甲生长发育及成虫越夏中起着多重作用。  相似文献   

2.
李玲  李爽  李娜  庞保平 《植物保护学报》2021,48(6):1447-1456
为明确沙葱萤叶甲Galeruca daurica钙结合蛋白基因的功能,利用本实验室组装的沙葱萤叶甲转录组数据库鉴定钙结合蛋白基因序列,采用RNA干扰（RNA interference,RNAi）技术沉默沙葱萤叶甲3龄幼虫体内钙调蛋白（calmodulin,CaM）、钙网蛋白（calreticulin,CRT）、类钙磷蛋白（calcyphosine-like,CAPSL）和类肌钙蛋白C（troponin C-like,TnCl）共4种钙结合蛋白基因,观察幼虫发育历期、存活率、体重及蛹重的变化;应用实时荧光定量PCR测定干扰效率。结果表明,共筛选到41条编码钙结合蛋白的基因序列,选取4条具有完整开放阅读框的钙结合蛋白基因序列进行后续研究。4种钙结合蛋白基因的干扰效率从高到低依次为CAPSL（94.4%）、TnCl（76.2%）、CRT（70.5%）和CaM（54.5%）;干扰效率最高的时间分别为干扰后第4、2、6和2天。沉默CAPSL后,沙葱萤叶甲幼虫体重显著降低了21.0%~34.9%,存活率显著下降了53.5%,发育历期显著缩短了15.1%,蛹重显著降低了15.8%。沉默TnCl后,沙葱萤叶甲幼虫体重显著降低了10.5%~25.0%,存活率显著下降了19.1%,蛹重显著降低了11.0%,而对发育历期无显著影响。沉默CaM后,沙葱萤叶甲幼虫体重显著降低了5.9%~6.6%,存活率显著降低了8.7%,而幼虫发育历期和蛹重无显著变化。沉默CRT后,沙葱萤叶甲幼虫体重、蛹重、存活率和发育历期均无显著变化。表明CAPSL、TnCl和CaM在沙葱萤叶甲幼虫生长发育过程中发挥着重要作用,而CRT可能未参与沙葱萤叶甲幼虫生长发育的调控过程。  相似文献   

3.
为探究丝氨酸蛋白酶抑制剂（serine protease inhibitor,Serpin）基因在黏虫Mythimna separata生长发育及免疫防御等过程中发挥的作用,在黏虫转录组中获得Serpin基因的cDNA序列,采用实时荧光定量PCR技术分析Serpin在黏虫不同发育阶段和不同组织中的表达水平,并通过RNA干扰技术分析Serpin在黏虫抗球孢白僵菌Beauveria bassiana过程中发挥的作用。结果显示,获得2个黏虫Serpin基因,命名为MsSerpin-4和Mserpin-5,其cDNA全长序列分别为1 938 bp和2 618 bp,开放阅读框分别为1 188 bp和1 197 bp,分别编码395个和398个氨基酸,GenBank登录号分别为MZ577062和MZ577063。2个基因在黏虫不同龄期和不同组织内均有表达,在成虫期的相对表达量最高,在1龄幼虫期的相对表达量最低;在中肠中的相对表达量最高,在唾腺中的相对表达量最低。RNA干扰6 h时对MsSerpin-4和Mserpin-5表达的抑制效果最好,抑制率分别为52.57%和49.15%;MsSerpin-4和Mserpin-5的表达被抑制后,黏虫对球孢白僵菌更敏感,校正死亡率分别提高了31.47%和14.62%。表明黏虫2个MsSerpin均参与了被球孢白僵菌侵染后的免疫反应,可为后续黏虫的生物防治提供新靶标。  相似文献   

4.
为明确小热激蛋白在沙葱萤叶甲Galeruca daurica应对温度胁迫中的作用,采用PCR方法克隆沙葱萤叶甲小热激蛋白基因Hsp20（GdHsp20.6）完整的开放阅读框（open reading frame,ORF）序列,应用在线软件对GdHsp20.6基因进行生物信息学分析,通过原核表达技术诱导表达及纯化其编码蛋白,并通过实时荧光定量PCR（real-time quantitative PCR,RT-qPCR）技术分析不同温度胁迫下GdHsp20.6基因的表达量。结果显示,GdHsp20.6基因ORF序列长度为543 bp,编码180个氨基酸,预测分子量为20.6 kD,无跨膜区和信号肽。GdHsp20.6氨基酸序列有高度保守的α-结构域。GdHsp20.6氨基酸序列与其它鞘翅目昆虫的Hsp20氨基酸序列有较高的一致性,其中与花绒寄甲Dastarcus helophoroides的Hsp20.99氨基酸序列的一致性最高,为63%。GdHsp20.6基因在大肠杆菌Escherichia coli BL21（DE3）细胞系中成功表达,经异丙基-β-D-硫代吡喃半乳糖苷（isopropyl-β-d-thiogalactoside,IPTG）诱导后GdHsp20.6蛋白成功表达,十二烷基硫酸钠-聚丙烯酰胺凝胶电泳（sodium dodecyl sulfate-polyacrylamide gel electrophoresis,SDS-PAGE）和Western-blot分析表明融合蛋白大小与预测大小一致,并纯化获得了纯度较高的目的蛋白GdHsp20.6。低温（-10~5℃）和高温（35~40℃）处理1 h以及处理后25℃恢复30 min均能诱导GdHsp20.6基因表达上调,并且0℃处理30~120 min也能诱导GdHsp20.6基因表达上调。表明GdHsp20.6基因在沙葱萤叶甲应对低温和高温胁迫中可能起着重要作用。  相似文献   

5.
为揭示沙葱萤叶甲Galeruca daurica表皮蛋白基因GdAbd在其应对温度胁迫中的作用,采用RACE技术克隆表皮蛋白基因GdAbd的cDNA全长序列,并进行生物信息学分析;利用实时荧光定量(qPCR)技术比较GdAbd经不同温度处理1 h及25℃恢复30 min后在沙葱萤叶甲2龄幼虫体内的表达水平。结果表明,GdAbd基因全长708 bp(GenBank登录号:MG874710),开放阅读框为477 bp,编码158个氨基酸;蛋白预测分子量为16.98 kD,等电点为4.26;编码蛋白具有典型的表皮蛋白RR保守结构域,属于RR-2亚族;具有1个跨膜结构和1个信号肽。同源序列比对和系统发育分析表明,GdAbd与马铃薯甲虫Leptinotarsa decemlineata SgAbd-4的同源性最高,氨基酸序列一致性达50.63%。qPCR检测结果表明,与25℃对照相比,GdAbd基因表达水平在-10～5℃低温胁迫时未发生显著变化,而在35℃高温胁迫时发生显著上调;-10、-5和0℃低温胁迫后25℃恢复30 min可诱导GdAbd显著上调表达,但5℃低温和35℃高温胁迫处理与对照差异不显著。说明短时低温胁迫不能显著影响GdAbd表达,但胁迫后回温可诱导其上调表达。  相似文献   

6.
为明确韭菜迟眼蕈蚊Bradysia odoriphaga紫外敏感视蛋白基因Bo-uv的作用及其与趋光性的关系,利用常规PCR方法克隆获得Bo-uv基因的全长cDNA序列,分析了其敏感视蛋白的氨基酸序列与其它12种昆虫同源蛋白氨基酸序列之间的系统进化关系,运用qPCR技术检测了不同发育阶段、不同组织及不同光强度下Bo-uv基因的相对表达量。结果表明,Bo-uv基因cDNA全长2 757 bp,开放阅读框1 542 bp,编码514个氨基酸。韭菜迟眼蕈蚊紫外敏感视蛋白的氨基酸序列与其它12种昆虫同源蛋白的氨基酸序列一致性为21.93%～43.00%,与橘小实蝇Bactrocera dorsalis的氨基酸序列同源性最高。Bo-uv基因在韭菜迟眼蕈蚊蛹末期、成虫期表达,在成虫头部的相对表达量较高。在0～10 000 lx光强范围内雌、雄成虫体内该基因的相对表达量均呈先增高后降低趋势。与对照相比,1 000 lx光强度下其相对表达量显著升高,10 000 lx时相对表达量显著降低。表明光强度能够有效地调控Bo-uv基因的表达,该基因在韭菜迟眼蕈蚊感知外界光刺激过程中具有重要作用。  相似文献   

7.
为明确N6-甲基腺苷（N6-methyladenosine,m6A）修饰在棉铃虫核型多角体病毒（Helicoverpa armigera nucleopolyhedrovirus,HearNPV）侵染棉铃虫中的作用,根据基因组和转录组数据,对棉铃虫m6A结合蛋白基因YTHDF1（YTH domain-containing family protein 1）进行鉴定和分析,利用实时荧光定量PCR技术检测棉铃虫YTHDF1基因的时空表达模式、HearNPV处理后的表达变化情况以及RNA干扰效率,并调查该基因下调后对HearNPV复制及感染HearNPV棉铃虫死亡情况的影响。结果显示,棉铃虫YTHDF1基因开放阅读框全长为2 019 bp,编码672个氨基酸,含有1个保守的YTH结构域,其氨基酸序列与斜纹夜蛾Spodoptera litura同源物序列一致性达87.52%。YTHDF1基因在棉铃虫不同发育阶段均有表达,其中在进入5龄期24 h幼虫中的表达量最高,在2龄取食期幼虫中的表达量最低。YTHDF1基因的空间表达谱呈现一定的组织特异性,在幼虫血细胞和成虫头部的表达量最高。HearNPV侵染棉铃虫后YTHDF1基因上调表达,经RNA干扰使该基因下调后显著抑制了HearNPV多角体蛋白基因polyhedrin的表达并延迟了感染HearNPV棉铃虫的死亡时间。表明YTHDF1基因在HearNPV侵染棉铃虫的过程中发挥着重要作用。  相似文献   

8.
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9.
为明确小热激蛋白在沙葱萤叶甲Galeruca daurica应对温度胁迫中的作用,采用PCR方法克隆沙葱萤叶甲小热激蛋白基因Hsp20(GdHsp20.6)完整的开放阅读框(open reading frame,ORF)序列,应用在线软件对GdHsp20.6基因进行生物信息学分析,通过原核表达技术诱导表达及纯化其编码蛋白,并通过实时荧光定量PCR(real-time quantitative PCR,RT-qPCR)技术分析不同温度胁迫下GdHsp20.6基因的表达量。结果显示,GdHsp20.6基因ORF序列长度为543 bp,编码180个氨基酸,预测分子量为20.6 kD,无跨膜区和信号肽。GdHsp20.6氨基酸序列有高度保守的α-结构域。GdHsp20.6氨基酸序列与其它鞘翅目昆虫的Hsp20氨基酸序列有较高的一致性,其中与花绒寄甲Dastarcus helophoroides的Hsp20.99氨基酸序列的一致性最高,为63%。GdHsp20.6基因在大肠杆菌Escherichia coli BL21(DE3)细胞系中成功表达,经异丙基-β-D-硫代吡喃半乳糖苷(isopropyl-β-d-thiogalactoside,IPTG)诱导后GdHsp20.6蛋白成功表达,十二烷基硫酸钠-聚丙烯酰胺凝胶电泳(sodium dodecyl sulfate-polyacrylamide gel electrophoresis,SDS-PAGE)和Western-blot分析表明融合蛋白大小与预测大小一致,并纯化获得了纯度较高的目的蛋白GdHsp20.6。低温(-10～5℃)和高温(35～40℃)处理1 h以及处理后25℃恢复30 min均能诱导GdHsp20.6基因表达上调,并且0℃处理30～120 min也能诱导GdHsp20.6基因表达上调。表明GdHsp20.6基因在沙葱萤叶甲应对低温和高温胁迫中可能起着重要作用。  相似文献   

10.
为探究过氧化物还原酶（peroxiredoxin,Prx）在棉铃虫Helicoverpa armigera幼虫体内的功能,利用转录组数据鉴定得到了棉铃虫Prx4基因,并对其进行生物信息学分析,采用实时荧光定量PCR（real-time quantitative polymerase chain reaction,qRT-PCR）测定该基因在棉铃虫不同发育阶段和5龄幼虫不同组织中的相对表达,利用原核表达和RNA干扰方法探究该基因的功能。结果表明,Prx4基因开放阅读框长为744 bp,编码248个氨基酸,系统进化树和同源序列多重比对结果显示该基因属于典型的2-Cys过氧化物酶家族;Prx4基因广泛分布于棉铃虫幼虫各组织和发育阶段中;当感染核型多角体病毒（nucleopolyhedrovirus,NPV）后,棉铃虫Prx4基因相对表达量上调;重组蛋白的体外试验表明Prx4蛋白具有一定的抗氧化活性;与对照相比,小干扰RNA处理6、8和10 d后感病棉铃虫的死亡率分别较对照显著提高了4.46%、22.42%和38.68%。表明Prx4基因作为一种抗氧化酶保护棉铃虫免受氧化损伤,同时在抵御NPV感染过程中发挥着重要的免疫作用。  相似文献   

11.
Fifteen species of dermestid beetles were recorded at ‘Evolution Canyon’ (EC), Lower Nahal Oren, Mt. Carmel, Israel. They represent ~35% of known Israeli dermestid species. The following three species were recorded for the first time in Israel:Trogoderma svriaca Dalla Torre, 1911;Ctesias svriaca Ganglbauer, 1904; andAnthrenus (s.str.) jordaniens Pic, 1934. Adults of 13 species were collected on the more solar radiated, warmer and climatically more fluctuating south-facing slope (SFS); ten species were collected on the opposite, north-facing slope (NFS), which was cooler and climatically more stable. The abundance of adult dermestid beetles was 1.9 times higher on the SFS than on the NFS (86 and 47, respectively). Species richness and abundance distribution at EC (three collecting stations on each slope and one at the valley bottom) were significantly negatively correlated with the plant cover that consisted of trees and bushes (Spearmanr s ,P=0.007 and 0.039, respectively) and perennials (Spearmanr s ,P=0.039 and 0.077, respectively), indicating that non-woody plants were preferred by adult dermestid beetles.  相似文献   

12.
Inoculum density, temperature, leaf age, and wetness duration were evaluated for their effects on the development of black streak (Itersonilia perplexans) on edible burdock (Arctium lappa L.) in a controlled environment. The effect of relative humidity (RH) on ballistospores production by I. perplexans was also evaluated. Symptoms of black streak on leaves increased in a linear fashion as the inoculum density of I. perplexans increased from 102 to 106 ballistospores/ml. Rugose symptoms on young leaves were observed at densities of ≥104 ballistospores/ml. Disease severity of I. perplexans in relation to leaf age followed a degradation curve when the leaves were inoculated with ballistospores. Disease severity was high in newly emerged leaves up to 5 days old, declined as leaf age increased to 29 days, and was zero when leaf age increased from 30 to 33 days. Disease development of edible burdock plants exposed to ballistospores of I. perplexans was evaluated at various combinations of temperature (10°, 15°, 20°, 25°C) and duration of leaf wetness (12, 24, 36, 48, and 72 h). Disease was most severe when plants were in contact with the ballistospore sources at 15° or 20°C. The least amount of disease occurred at 25°C regardless of wetness duration. Ballistospores required 24–36 h of continuous leaf wetness to cause visible symptoms by infection on edible burdock. Ballistospores production in infected lesions required at least 95.5% RH.  相似文献   

13.
Bactra venosana Zeller (Lepidoptera: Tortricidae) is the predominant natural enemy ofCyperus rotundas L. in Bangalore, India. Although the insect was found to have potential, it was incapable of suppressing the weed due to low incidence. Laboratory studies involving field-collected insect stages revealed that heavy parasitism of eggs byTrichogrammatoidea bactrae Nagaraja (Hymenoptera: Trichogrammatidae) was the most important mortality factor suppressing its potentiality.  相似文献   

14.
Two Fusarium species, F. oxysporum f.sp. asparagi and F. proliferatum, are known to be involved in the root and crown rot complex of asparagus. We have investigated reports on the involvement of F. redolens, a third species, which until recently was considered conspecific with F. oxysporum because of morphological similarities. RFLP analysis of the rDNA internal transcribed spacer region and AFLP fingerprinting identified eight strains from asparagus unambiguously as F. redolens. Four of these were tested and found to be pathogenic to asparagus either in this study (two strains) or in a previous one in which they were classified as F. oxysporum (three strains). Disease symptoms and disease development were the same as with F. oxysporum f.sp. asparagi and F. proliferatum. Present data and literature reports identify F. redolens as a host-specific pathogen involved in root, crown and spear rot of asparagus. The pathogen is formally classified as F. redolens Wollenw. f.sp. asparagi Baayen.  相似文献   

15.
Pseudothecia ofMycosphaerella graminicola, the teleomorph ofSeptoria tritici, were observed on leaves of winter wheat (Triticum aestivum L.) in Slovakia for the first time in 2004. The samples of this pyrenomycetous ascomycete were compared with previously published data by micrometric characteristics using light microscope observations of morphological characteristics. http://www.phytoparasitica.org posting July 18, 2005.  相似文献   

16.
Approximately 30,000 fluorescent bacterial strains isolated from tomato, lettuce, eggplant, Chinese cabbage, and Japanese pepper plants at seven different locations in Hyogo Prefecture, were screened for plant-growth-promoting (PGP) activity to induce disease resistance against bacterial wilt in tomato. The 37 strains that had higher PGP activity were subjected to molecular phylogenetic analyses using the sequences of the 16S rRNA, gyrB and rpoD genes. Most of the strains were identified as Pseudomonas fluorescens or its close relative, P. putida, while a few strains were grouped with more distantly related bacterial species such as Enterobacter and Stenotrophomonas. The phylogenetic relationships among tomato and lettuce isolates mostly coincided with the source locality and host plants, with a few exceptions. In contrast, isolates from Japanese pepper plants did not form their own cluster but represented several different bacterial species.  相似文献   

17.
Cereal aphids on a wheat crop were sampled through the winter of 1982/1983 using both fixed and random quadrats.Sitobion avenae overwintered successfully anholocyclically although there was a substantial decline in the population of this species. No evidence for successful anholocyclic overwintering in other aphid species was found, althoughRhopalosiphum padi was seen to colonise the crop in early winter and was found until the end of December.Eggs ofR. padi onPrunus padus were observed through the winter. A steady decline in their numbers occurred. The overall survival rate of the 5385 eggs was c.30%.Eggs ofMetopolophium dirhodum onRosa spp. were also monitored. They also showed a steady decline in numbers through the winter. The overall survival rate of the 1360 eggs was c.34%.Egg hatch in bothR. padi andM. dirhodum was closely synchronised with bud burst of their host plant. For the latter species this resulted in egg hatch starting in January.Samenvatting In de winter van 1982/1983 werden de graanluizen in een tarwegewas regelmatig geteld in willekeurig en in van tevoren gekozen monstereenheden. Hoewel er een aanmerkelijke afname van de populatie vanSitobion avenae werd waargenomen, overwinterde deze soort toch anholocyclish. Er werden geen aanwijzingen gevonden dat de andere bladluissoorten anholocyclish overwinterden hoewel kolonies vanRhopalosiphum padi tot eind december werden waargenomen.De gehele winter werden eieren vanR. padi opPrunus padi aangetroffen, de aantallen ervan namen geleidelijk af; van de 5385 getelde eieren overleefde ongeveer 30%. Ook de aantallen eieren vanMetopolophium dirhodum opRosa spp. werden reglematig vastgesteld; ook hier werd een geleidelijke afname geconstateerd. Van de oorspronkelijk getelde 1360 eieren overleefde ongeveer 34%.Het uitkomen van de eieren vanR. padi enM. dirhodum viel nauw samen met het uitlopen van de knoppen van hun waardplant. VoorM. dirhodum betekende dit dat de eerste eieren al begin januari uitkwamen.  相似文献   

18.
采用生物活性追踪法,从苦楝根皮、树皮和果实中分离出多种四环三萜类杀虫有效成分。这些成分对菜青虫和亚洲玉米螟均表现出明显的拒食活性,川楝素对菜肯虫还有明显的胃毒活性。从苦楝中分离出的几种非四环三萜类物质对昆虫的生物活性均较低,仅表现出一定的拒食活性。另外对几种三萜衍生物的生物活性进行了测定与比较。  相似文献   

19.
Leaves ofPelargonium domesticum andRuscus hypoglossum infected byBotrytis cinerea Pers., produced 12 and 1.5 nl ethylene/h/g, respectively, 34 days after inoculation; wounded or healthy leaves and phyloclades of them produced much lower amounts. When the fungus was grown on dead leaves it produced negligible amounts of the gas even when supplemented with methionine. Exogenous ethylene enhanced gray mold development in both hosts. Silver thiosulfate, aminooxyacetic acid (AOA) and aminoethoxyvinylglycine significantly inhibited disease development in pelargonium, and the latter two compounds inhibited ethylene production. AOA inhibited disease development and ethylene production in cut rose flowers; calcium ions inhibited disease development whereas the chelator EGTA [ethylene glycol bis-(β-aminoethyl ether)N,N,N′,N′-tetraacetic acid] enhanced it. Disease suppression by an excess of Ca2+ was correlated with repression of ethylene production by the flowers, whereas deficiency in Ca2+ increased disease severity. Publication of the Agricultural Research Organization. No. 2413-E, 1988 series.  相似文献   

20.
In the present study,Cnaemidophorus rhododactyla (Denis & Schiffermüller) (Lepo Pterophoridae) was noted for the first time as a pest ofRosa spp. in Turkey. The larva ofC. rhododactyla makes a cavity by feeding in the flower-bud and destroys at least 60% of the mass of the flower-bud. There are approximately 12 species ofRosa in Erzurum and Kars provinces, butRosa dumalis Bechst. andR. subcanina L. suffered more damage than the otherRosa species. One species of Tachinidae (Diptera) —Pseudoperichaeta palesoidea (Robineau-Desvoidy), and two species of Ichneumonidae (Hymenoptera) —Sinophorus turionus (Ratz.) andScambus brevicornis Gravenhorst, were reared as larval parasitoids ofC. rhododactyla. C. rhododactyla is a new host record for these parasitoids. Additionally,S. turionus is a new record for the Turkish fauna.  相似文献   

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