四种Bt毒素蛋白对二点委夜蛾幼虫毒力测定及中肠组织病理学变化

为更好地了解苏云金芽胞杆菌Bacillus thuringiensis毒素蛋白对二点委夜蛾Athetis lepigone的毒力以及作用机理,通过饲喂含有Cry1Ac、Cry1Ab、Cry2Ab和Vip3Aa四种不同Bt毒素蛋白饲料,测定Bt毒素蛋白对二点委夜蛾幼虫的毒力,并观察取食4种毒素蛋白后幼虫中肠组织的病理学变化。结果显示,二点委夜蛾幼虫取食毒素蛋白后72 h,Cry1Ab和Cry1Ac毒素蛋白对二点委夜蛾幼虫的杀虫活性较高,校正死亡率为84.7%和76.4%;Vip3Aa和Cry2Ab毒素蛋白的毒力较弱。二点委夜蛾幼虫取食4种Bt毒素蛋白后,中肠柱状细胞微绒毛脱落,杂乱地分散在肠腔内,杯状细胞变形和腔内微绒毛脱落,线粒体和内质网等变形破裂,细胞核的核膜消失、核质凝聚和形状发生变化,经Cry1Ab和Cry1Ac毒素蛋白处理后中肠细胞的病变症状和速度明显高于Cry2Ab和Vip3Aa毒素蛋白处理。表明Cry1Ab和Cry1Ac毒素蛋白对二点委夜蛾幼虫杀虫活性较高,显著高于Cry2Ab和Vip3Aa毒素蛋白,且对其中肠细胞的破坏作用也较强。     

小菜蛾中肠氨肽酶N2在昆虫细胞中的表达

利用昆虫细胞-杆状病毒表达系统(Bac-to-Bac baculovirus expression system)表达获得小菜蛾 Plutella xylostella(L.)中肠氨肽酶N2(aminopeptidase N,APN2)蛋白,成功建立了该蛋白在昆虫细胞中的表达体系。将对Cry1Ac毒素敏感和已产生抗性的小菜蛾中肠APN2基因插入表达载体pFAST Bac HTB中,并在草地夜蛾 Spodoptera frugiperda 细胞系(Sf9)中进行了重组表达。聚丙烯酰胺凝胶电泳(SDS-PAGE)及蛋白免疫印迹技术(Western-blotting)结果显示,在107 kDa处有1条特异性蛋白条带。研究表明,小菜蛾中肠APN2基因可在Sf9细胞中成功表达,这为继续研究其功能及抗性的关系奠定了基础。     

丁布胁迫对亚洲玉米螟氧化还原系统的影响

为探明丁布胁迫对亚洲玉米螟氧化还原系统的影响,采用酶活力测定方法和实时荧光定量PCR技术,研究了取食丁布含量差异显著的3个玉米品种登海662、浚单20和隆玉602叶片及模拟其叶片丁布含量的人工饲料后,玉米螟3龄幼虫氧化还原系统主要酶活性和谷氧还蛋白(Grx)mRNA转录表达的变化。结果显示:取食3个玉米品种叶片24 h后,亚洲玉米螟幼虫过氧化物酶和超氧化物歧化酶活性均显著高于对照,且在3个品种处理间差异显著,由大到小依次为登海662(118.30 U/mg和182.19 U/mg)、浚单20(107.13 U/mg和90.02 U/mg)和隆玉602(77.54 U/mg和93.59 U/mg)。取食浚单20可显著促进幼虫脂肪体Grx的表达,取食登海662可显著促进其中肠Grx的表达;取食模拟登海662叶片丁布含量的人工饲料12 h后,幼虫脂肪体中Grx表达量高于对应玉米叶片饲喂处理,而中肠部位则低于对应玉米叶片饲喂处理;取食模拟浚单20的结果与模拟登海662相反。表明亚洲玉米螟氧化还原系统在防御丁布胁迫过程中发挥着重要作用。     

Pathogenicity of fungi affecting black locust(Robinia pseudoacacia) in Greece

The main fungi affecting black locust(Robinia pseudoacacia L.) were recorded during a survey which took place in northern Greece in 1997. It was found thatPhomopsis oncostoma (Thüm.) v. Höhn,Aglaospora profusa (Fr.) de Not. andCucurbitaria elongata (@#@ Fr.) Grev. cause necroses of twigs, branches and even entire trees. The pathogenicity of these fungi was confirmed by inoculation experiments in which typical disease symptoms appeared with re-isolation of the respective fungi from the inoculated plants.     

Effect of different hosts onThaumetopoea pityocampa populations in northeast Portugal

A study was carried out at the Natural Park of Montesinho, NE Portugal, in order to evaluate the effect of different pine species (Pinus pinaster Ait.,P. nigra Arn. andP. sylvestris L.) onThaumetopoea pityocampa populations. The structure of the egg batches, the impact of the egg parasitoids on natural mortality of the pest and the species of parasitoids present, as well as their emergence dynamics, were analyzed. The length of the egg batches varied among pine species with the longest ones onP. nigra. The mean number of eggs per batch differed betweenP. sylvestris and the two other hosts studied, with fewer eggs per batch on the first. No differences were found in the size of eggs among pine species. The egg mortality varied between 25.8% and 33.0%, with no differences among hosts. Parasitism was the main cause of death.Baryscapus servadeii (Mercet.) was the most abundant parasitoid species, followed byOoencyrtus pityocampae (Dom.) andTrichogramma embryophagum Htg.B. servadeii dominated in the egg batches collected fromP. pinaster andP. nigra, whereasO. pityocampae was most frequent onP. sylvestris. The emergence ofB. servadeii started in the middle of March and continued until August, with the emergence peak at the end of May. The emergence ofO. pityocampae started at the end of April and continued throughout September, with maximum values in June. http://www.phytoparasitica.org posting Sept. 20, 2006.     

草地贪夜蛾对金银花的取食适应性

为明确新入侵农业害虫草地贪夜蛾Spodoptera frugiperda对药用植物金银花Lonicera japonica的潜在为害风险,采用室内饲养观察法比较取食玉米Zea mays和金银花叶片后草地贪夜蛾的种群特征,并采用室内生测法测定取食2种植物幼虫体内消化酶和解毒酶的活性。结果显示,草地贪夜蛾在金银花上能完成完整的世代发育。取食金银花叶片的草地贪夜蛾幼虫发育历期为24.08 d,显著长于取食玉米的幼虫发育历期（18.47 d）;取食金银花叶片的幼虫存活率为63.67%,较取食玉米叶片幼虫的存活率78.00%明显降低;取食金银花叶片的草地贪夜蛾雌、雄蛹重分别为197.98 mg和198.78 mg,显著大于取食玉米叶片时;取食金银花和玉米叶片的草地贪夜蛾成虫的繁殖力与寿命均无显著差异。取食金银花叶片的草地贪夜蛾5龄幼虫中肠中的淀粉酶、脂肪酶、胰蛋白酶、谷胱甘肽S-转移酶和细胞色素P450酶的活性均较取食玉米叶片时显著降低,但羧酸酯酶活性较取食玉米叶片时显著升高。表明草地贪夜蛾在金银花上能较好地生长发育和繁殖,这对金银花的安全生产构成潜在威胁。     

Antagonistic activity of endophytic fungi towards <Emphasis Type="Italic">Diplodia corticola</Emphasis> assessed by in vitro and in planta tests

One isolate each of Trichoderma viride, Epicoccum nigrum, Fusarium tricinctum, Alternaria alternata, Sclerotinia sclerotiorum and Cytospora (teleomorph: Valsa sp.) present in epigeous declining oak tissues was evaluated for its ability to control Diplodia corticola (isolate 79). This fungus is the causal agent of cankers, vascular necrosis and dieback on various oak species. Among the isolates tested, T. viride and F. tricinctum showed maximum in vitro inhibition of mycelial growth of D. corticola (isolate 79). Species were also evaluated for their ability to reduce mortality caused by D. corticola (isolate 79) of Quercus cerris and Q. pubescens seedlings under controlled conditions. Two series of inoculations were carried out through wounds in the stem; in the first, the distance between the point of inoculation of the antagonist and the pathogen was 6 cm, whereas in the second series the distance was shortened to 3 cm. In seedlings of Q. cerris and Q. pubescens at a distance of 3 cm, inoculation with F. tricinctum and A. alternata significantly reduced mortality caused by D. corticola (isolate 79). Inoculation of T. viride through artificial cuticular wounds in the stem of seedlings prevented the proliferation of D. corticola (isolate 79) only on seedlings of Q. cerris. All Q. pubescens seedlings treated with T. viride manifested pathological symptoms subsequent to proliferation of D. corticola (isolate 79). These observations indicate that the interactions between endophytes in planta and D. corticola (isolate 79) are complex and merit further study.     

The oak gallwasp <Emphasis Type="Italic">Aphelonyx persica:</Emphasis> a new record from Turkey,with some new host records

A new record is reported of the oak gallwasp Aphelonyx persica on oaks (Quercus cerris, Q. ithaburensis, Q. trojana) in the Inland Aegean Region of Turkey. This species is recorded for the first time on Q. cerris and Q. trojana. Details are given of the geographical distribution, hosts and phenology.     

转Cry1Ac/1Ab基因棉花对棉铃虫的控制效果及对甜菜夜蛾和斜纹夜蛾生长发育的影响

为评估转Cry1Ac/1Ab基因棉花对棉铃虫Helicoverpa armigera的抗性及对非靶标害虫甜菜夜蛾Spodoptera exigua和斜纹夜蛾S.litura生长发育的影响,采用室内生测法测定其对棉铃虫的抗性及对甜菜夜蛾和斜纹夜蛾不同龄期幼虫存活率、营养代谢及中肠酶活性的影响。结果表明,转Cry1Ac/1Ab基因棉花对棉铃虫第2代幼虫的抗性程度最高,幼虫校正死亡率达91.33%,但对甜菜夜蛾和斜纹夜蛾的抗性程度较低,幼虫校正死亡率分别为15.33%和13.33%。甜菜夜蛾和斜纹夜蛾各龄期幼虫取食转Cry1Ac/1Ab基因棉花后,其存活率与取食常规棉花对照无显著差异;甜菜夜蛾对转Cry1Ac/1Ab基因棉花叶片的相对取食量、近似消化率分别为16.68和93.12%,均高于取食常规棉花对照的10.72和92.00%,但差异不显著,而斜纹夜蛾取食转Cry1Ac/1Ab基因棉花后的各项营养指标均低于取食常规棉花对照,差异也不显著。取食转Cry1Ac/1Ab基因棉花后,甜菜夜蛾的过氧化物酶活性为0.02 U/mg prot,显著低于取食常规棉花的0.05 U/mg prot;斜纹夜蛾的酸性磷酸酶活性为0.15 U/mg prot,高于取食常规棉花的0.10 U/mg prot,但差异不显著,其它中肠酶活性均低于对照,亦无显著差异。     

Note: Life history of<Emphasis Type="Italic">Lixus bardanae</Emphasis> on curly dock (<Emphasis Type="Italic">Rumex crispus</Emphasis>) in Turkey

The biology ofLixus bardanae (F.) (Coleoptera: Curculionidae) on curly dock (Rumex crispus L.) in northeastern Anatolia (Bayburt, Erzurum and Kars Provinces), Turkey, was studied during the years 2000 and 2001.L. bardanae completes one generation in a year, overwintering as an adult. It feeds on leaves of the host plant. Females lay eggs individually into stems and the young larvae create galleries in stems while feeding. Pupation occurs inside stems in cells fashioned from frass. In late September, adults move into soil and overwinter in an upright position around the roots of the host plant. Infestation levels were found to range between 34% and 84%. Two parasitoids,Exeristes roborator F. andEndromopoda phragmitidis Perve (Hymenoptera: Ichneumonidae), were reared fromL. bardanae. http://www.phytoparasitica.org posting Dec. 21, 2003.     

First report of Rhizomonas sp. causing corky root of lettuce in Europe

Typical symptoms of corky root were observed on iceberg lettuce (Lactuca sativa L.) in the Netherlands and England, on prickly lettuce (Lactuca serriola L.) in Spain, and on sowthistle (Sonchus oleraceus L.) in Greece. Slow-growing bacteria with similar colonies as strains ofRhizomonas suberifaciens or otherRhizomonas species were isolated from soil surrounding plants with corky root symptoms using lettuce seedlings as bait. Crude lysate from all strains was tested for DNA homology with DNA fromR. suberifaciens strain CA1 (R. sub. CA1) andRhizomonas sp. strain WI4 (R. sp. WI4). Strains that had homology values higher than that ofR. sp. WI4 orR. sub. CA1 were tested for pathogenicity on 1-wk-old lettuce seedlings, cv. Salinas, Two strains from the Netherlands induced typical symptoms of corky root on lettuce. These strains tested negative with monoclonal antibody MAb-Rs1 specific forRhizomonas suberifaciens in an enzyme-linked immunosorbent assay (ELISA). They had low DNA homology withR. sub. CA1 (4–9%) and low to moderate DNA homology withR. sp. WI4 (4–17%). Some nonpathogenic strains had moderate to high levels of DNA homology withR. sub. CA1 orR. sp. WI4 (19–84%). All strains had fatty acid profiles similar to those ofRhizomonas species. This is the first report ofRhizomonas sp. causing corky root of lettuce in Europe.     

Cytological events in tomato leaves inoculated with conidia of <Emphasis Type="Italic">Blumeria graminis</Emphasis> f. sp. <Emphasis Type="Italic">hordei</Emphasis> and <Emphasis Type="Italic">Oidium neolycopersici</Emphasis> KTP-01

Leaves of tomato and barley were inoculated with conidia of Blumeria graminis f. sp. hordei race 1 (R1) or Oidium neolycopersici (KTP-01) to observe cytological responses in search of resistance to powdery mildew. Both conidia formed appressoria at similar rates on tomato or barley leaves, indicating that no resistance was expressed during the prepenetration stage of these fungi. On R1-inoculated tomato leaves, appressoria penetrated the papillae, but subsequent haustorium formation was inhibited by hypersensitive necrosis in the invaded epidermal cells. On the other hand, KTP-01 (pathogenic to tomato leaves) successfully developed functional haustoria in epidermal cells to elongate secondary hyphae, although the hyphal elongation from some conidia was later suppressed by delayed hypersensitive necrosis in some haustorium-harboring epidermal cells. Thus, the present study indicated that the resistance of tomato to powdery mildew fungi was associated with a hypersensitive response in invaded epidermal cells but not the prevention of fungal penetration through host papilla.     

Note: Occurrence ofMycosphaerella graminicola, teleomorph ofSeptoria tritici, in Slovakia

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.     

美洲棉铃虫幼虫腹侧可外翻腺分泌物对番茄抗虫反应的影响

为明确美洲棉铃虫Helicoverpa zea幼虫腹侧可外翻腺(ventral eversible gland,VEG)分泌物对植物抗虫反应的影响,在宾夕法尼亚州立大学帕克分校校园日光温室大棚内分别测定取食未损伤、机械损伤后涂抹磷酸缓冲盐溶液(phosphate buffer saline,PBS)和机械损伤后涂抹VEG分泌物3种处理的番茄叶片后美洲棉铃虫幼虫体重增加量,不同处理番茄叶片中防御关键酶多酚氧化酶(polyphenol oxidase,PPO)活性、胰蛋白酶抑制剂(trypsin proteinase inhibitors,TPI)抑制率以及防御相关的蛋白酶抑制剂Ⅱ(proteinase inhibitor Ⅱ,PIN2)基因相对表达量。结果显示,3种处理中,取食机械损伤后涂抹VEG分泌物的番茄叶片的美洲棉铃虫幼虫体重较处理前增加量最少,为4.27 mg,显著低于其它2种处理;机械损伤后涂抹VEG分泌物的番茄叶片中PPO活性和TPI抑制率最高,分别为109.16 U/mg和27.48%,分别显著高于其它2种处理;同时机械损伤后涂抹VEG分泌物的番茄叶片中PIN2基因相对表达量显著上调,分别是未损伤和机械损伤后涂抹PBS缓冲液处理的11.83倍和1.17倍。表明美洲棉铃虫幼虫VEG分泌物能够促使防御关键酶的大量积累,使得防御相关基因的相对表达量上调,显著诱导番茄的抗虫反应,阻止后续昆虫的取食。     

Relationships of aphid and mite infestations to control ofBotrytis cinerea byClonostachys rosea in rose(Rosa hybrida) leaves

Infestations of aphids(Macrosiphum rosae L.) and of twospotted spider mites(Tetranychus urticae Koch) were examined in relation to growth and sporulation ofClonostachys rosea andBotrytis cinerea, and to suppression of the pathogen by the agent, in green rose leaves. Leaves were infested artificially with 10 aphids/leaflet for 3 h, or naturally with 15-30 aphids/leaflet for 7-12 days or with undetermined numbers of mites for 10-12 days. Leaves that had or had not been infested were inoculated withC. rosea, withB. cinerea, or withC. rosea plusB. cinerea. Germination incidence and germ tube growth ofC. rosea andB. cinerea on the phylloplane in most instances were much greater in leaves previously infested with aphids or mites compared with noninfested leaves. After combined inoculation,C. rosea suppressed germination ofB. cinerea from 47% to 19% in noninfested leaves, but in leaves that had been infested the agent was ineffective and germination incidence of the pathogen increased to 75-93%. Previous infestation with naturally introduced aphids or mites, but not brief infestations of artificially introduced aphids, markedly increased sporulation ofC. rosea after the leaves died during an initial 7-15 days of incubation on a paraquat agar medium, regardless of whether or notB. cinerea was present. Sporulation ofB. cinerea was similarly increased when inoculated alone. After 15-20 days, however, conidiophores of the agent or pathogen covered most of the leaf surface in these treatments. In leaves inoculated withC. rosea plusB. cinerea, the agent suppressed sporulation of the pathogen almost completely in both previously infested and noninfested leaves. Thus, aphid and mite infestations did not compromise the ability ofC. rosea to suppress inoculum production byB. cinerea in the leaves. Increased nutrient availability on the phylloplane through exudation or as honeydew or frass is proposed as a basis to explain effects of the pest infestations onC. rosea andB. cinerea.     

First report of leaf blight disease of <Emphasis Type="Italic">Gloriosa superba</Emphasis> L. caused by <Emphasis Type="Italic">Alternaria alternata</Emphasis> (Fr.) Keissler in India

Leaf blight disease was found on Gloriosa superba L. (Liliaceae), an endangered, herbaceous, perennial, climbing lily that produces colchicine, in West Bengal, India in 2004. Small brownish spots on leaves developed into concentric rings, which eventually darkened and coalesced to blight the entire leaf. The causal fungus was morphologically identified as Alternaria alternata (Fr.) Keissler. This is the first record of A. alternata on G. superba.     

Effect of storage on the efficacy of powdered leaves ofAnnona squamosa for the control ofCallosobruchus maculatus on cowpeas(Vigna unguiculata)

Annona squamosa L. is widespread in Sri Lanka and its leaves are collected by some farmers to protect their stored cowpeas from predation by bruchids. Removal of the leaves during the dry season, when the fanners harvest their cowpeas, can decrease the yield ofA. squamosa fruit. This study investigated whether storing the leaves before they are used would reduce the insecticidal activity of the leaves. Overall, acetone and ethanol extracts made from fresh and stored leaves ofA. squamosa decreased the number of adultCallosobruchus maculatus (F.) emerging from cowpeas; however, in some bioassays the extracts from fresh leaves were more active than those from leaves stored for 6 months. Acetone extracts from fresh and stored leaves were toxic to adult beetles, whereas the ethanol extracts were not active. Acetone and ethanol extracts from fresh leaves had potent ovicidal activity when applied to 2-day-old eggs.     

First report on the flower-rot of Crossandra infundibuliformis, a commercial flower crop in India

Crossandra infundibuliformis (L.) Nees is an important commercial plant in many cities in India, which is useful for bouquets, garlands, and decorations. The crop is affected by a new flower rot caused by Fusarium pallidoroseum (Cooke) Sacc. Severely infected plants had purplish leaves, decayed flowers and shortened internodes and were colonized by the cottony colonies of the fungus. The fungus was isolated and caused the same symptoms on plants after inoculation.     

Differential Responses to Pea Bacterial Blight in Stems, Leaves and Pods Under Glasshouse and Field Conditions

Resistance to pea bacterial blight (Pseudomonas syringae pv. pisi) in different plant parts was assessed in 19 Pisum sativum cultivars and landraces, carrying race-specific resistance genes (R-genes) and two Pisum abyssinicum accessions carrying race-nonspecific resistance. Stems, leaves and pods were inoculated with seven races of P. s. pv. pisi under glasshouse conditions. For both race-specific and nonspecific resistance, a resistant response in the stem was not always associated with resistance in leaf and pod. Race-specific genes conferred stem resistance consistently, however, there was variability in the responses of leaves and pods which depended on the matching R-gene and A-gene (avirulence gene in the pathogen) combination. R2 generally conferred resistance in all plant parts. R3 or R4 singly did not confer complete resistance in leaf and pod, however, R3 in combination with R2 or R4 enhanced leaf and pod resistance. Race-nonspecific resistance conferred stem resistance to all races, leaf and pod resistance to races 2, 5 and 7 and variable reactions in leaves and pods to races 1, 3, 4 and 6.Disease expression was also studied in the field under autumn/winter conditions. P. sativum cultivar, Kelvedon Wonder (with no R genes), and two P. abyssinicum accessions, were inoculated with the most frequent races in Europe under field conditions (2, 4 and 6). Kelvedon Wonder was very susceptible to all three races, whereas P. abyssinicum was much less affected. The combination of disease resistance with frost tolerance in P. abyssinicum enabled plants to survive through the winter. A breeding strategy combining race-nonspecific resistance derived from P. abyssinicum with race-specific R-genes should provide durable resistance under severe disease pressure.     

白菜型油菜BraSAUR基因家族鉴定及其表达分析

SAUR(Small auxin-up RNA)是生长素早期响应逆境胁迫的基因,本文研究白菜型油菜全基因组中SAUR家族的信息,分析了该家族成员的基本特征。以强抗寒性白菜型冬油菜品种陇油7号（L7）和弱抗寒性冬油菜品种陇Lenox（X）为试验材料,采用低温和干旱处理,应用荧光定量技术分析不同基因在不同品种中的表达特性,筛选差异表达基因,为研究生长素早期应答基因调控白菜型冬油菜生长点发育机理提供支撑。结果表明：白菜型油菜的BraSAUR基因共有142个,在10条染色体中不对称分布,主要分布在2、3号染色体上,基因长度较短,且大多数基因没有内含子。qRT-PCR结果表明,低温处理后,与CK相比,在两个品种的叶片中,Bra029452的表达量变化趋势不同,24 h时在L7中的表达量是X的24倍,在生长点中,Bra010501在L7中24 h时表达量达到CK的4倍,而在X中是先增加后降低的趋势。模拟干旱胁迫后,Bra029452在L7的叶片中的表达量均显著高于CK,在X中则是逐渐降低,在L7的生长点中1 h和24 h表达量均高于CK,但在X中则表现为先降低后升高。低温胁迫和干旱胁迫条件下,Bra029452基因表达量在强抗寒性油菜品种L7的叶片及生长点中均逐渐升高,可推断该基因同时参与白菜型冬油菜耐受低温、干旱的调控过程。