IPM in the transgenic era: a review of the challenges from emerging pests in Australian cotton systems

The Cotton Catchment Communities Cooperative Research Centre began during a period of rapid uptake of Bollgard II® cotton, which contains genes to express two Bt proteins that control the primary pests of cotton in Australia, Helicoverpa armigera and H. punctigera. The dramatic uptake of this technology presumably resulted in strong selection pressure for resistance in Helicoverpa spp. against the Bt proteins. The discovery of higher than expected levels of resistance in both species against one of the proteins in Bollgard II® cotton (Cry2Ab) led to significant re-evaluation of the resistance management plan developed for this technology, which was a core area of research for the Cotton CRC. The uptake of Bollgard II® cotton also led to a substantial decline in pesticide applications against Helicoverpa spp. (from 10–14 to 0–3 applications per season). The low spray environment allowed some pests not controlled by the Bt proteins to emerge as more significant pests, especially sucking species such as Creontiades dilutus and Nezara viridula. A range of other minor pests have also sporadically arisen as problems. Lack of knowledge and experience with these pests created uncertainty and encouraged insecticide use, which threatened to undermine the gains made with Bollgard II® cotton. Here we chronicle the achievements of the Cotton CRC in providing the industry with new knowledge and management strategies for these pests.

Evaluación de seis variedades de tomate (Lycopersicum esculentum Mill),bajo un manejo Mip, para el complejo mosca blanca geminivirus, en el Valle de Sébaco

El cultivo de tomate es una de las hortalizas mas preferidas para consumo fresco, las altas poblaciones de mosca blanca han provocado que los agricultores realicen excesivas aplicaciones de Insecticidas quimicos los cuales provocan efectos colaterales. En busca de métodos alternativos para el manejo de la mosca blanca geminivirus se planteó este trabajo para evaluar seis variedades de tomate lndustrial: milano, Silverados, FMX-922, NH-4764,NHW-785 y UC-82 B, a través de un B.C.R. con seis réplicas,donde los resultados muestran que la mayor incidencias de mosca blanca se presentó de (10-20) días después del transplante d.d.t. en todas las variedades. Cuando se realizó el análisis de insectos acumulados no se encontró diferencias entre las variedades.La Incidencias de virosis resultó significativas en las fechas 24 y 37 d.d.t. El rendimiento comercial de tomate para todas las variedades fue similar, sobresaliendo la variedad NH-4764.EI daño de frutos por Heliothis sp fue mayor en la variedad testigo UC-82 B,las variedades Silverados,milanos,FMX-922,NH-4764 y NHW-785, presentaron los porcentajes de daño más bajo con respecto al testigo, el daño de fruto por enfermedades fungosas fué medido en dos momentos 67 y 68 ddt, presentando NHW-785 el porcentaje más alto 44.6% en la primer fecha 67 ddt, en la segunda fecha 78 ddt la variedad FMK-922 fue la que presento el porcentaje mas alto.

Utilización de "microorganismos efectivos" en el manejo de insectos-plagas del cultivo del tomate (Lycopersicum esculentum Mill) variedad UC-82 en el Valle de Sébaco, Matagalpa

El trabajo se realizó de Octubre de 1996 a Marzo de 1997, en la Estación Experimental "Raúl González" del valle de Sébaco, Matagalpa. Se evaluó la utilización de "Microorganismos Efectivos" en el manejo de insectos-plagas y enfermedades en el cultivo del tomate (Lycopersicum esculentum Mili), variedad UC-82. Los tratamientos utilizados fueron: Tratamiento 1, Productos Químicos; Tratamiento 2, Neem (Azadirachta indica); Tratamiento 3, EM-5B (extracto de hierbas y microorganismos); Tratamiento 4, EM-5 (microorganismos); Tratamiento 5, Testigo absoluto (sin ningún tipo de aplicación). En la etapa de semillero los tratamientos EM-5 y EM-5B mostraron un efecto reductor sobre las poblaciones de mosca blanca (Bemisia tabaci Genn). Después del trasplante la mayor densidad de mosca blanca se observó en el tratamiento a base de productos químicos. El mayor porcentaje de poblaciones de Lyriomiza spp. lo obtuvieron los tratamientos químicos y EM-5. En cuanto al porcentaje de población de Diabrotica spp. no se presentaron diferencias significativas a lo largo del experimento. Referente al daño causado por Heliothis spp. se observó a los 77 DDT (días después del trasplante), los tratamientos en los que menos daños se presentaron fueron neem y químico. Sin embargo estos mismos tratamientos mostraron mayor incidencia de virosis a los 42 DDT, asi mismo no se observó diferencias estadísticas. La mayor altura la presentó el tratamiento EM-5B y el mayor número de hijos lo obtuvo el tratamiento químico y EM-5. También se presentaron resultados positivos entre los caracteres grosor del mesocarpio y diámetro polar y ecuatorial obteniendo los mejores resultados EM- 5 y EM-5B. En cuanto al rendimiento comercial de producción el tratamiento que dio valores más aceptables fue el de Neem con 26,974 Kg/Ha de frutos comercializables con rendimiento total 27,979 Kg/Ha; haciendo la salvedad que este tratamiento fue fertilizado con el biofertilizante EM-Bokashi

Período crítico de protección contra plagas del follaje y frutos del cultivo de la sandia (Citrullus vulgaris sugar baby) de exportación

Se hizo un experimento para determinar el efecto de periodos de protección de plagas del follajes y frutos sobre el rendimiento y la ganancia neta en el cultivo de sandía para exportación. el experimento fue ubicado entre Nagarote y la paz centro, realizándose en los meses de febrero a abril de 1988, bajo riego. Para la protección del cultivo de las plagas se utilizó el insecticida prfenofos más riopcord (tambo 880 ecu). La dinámica poblacional mostró que las plagas que provocaron daños directo a los frutos como larvas de spodoptera spp. (lepidóptera: noctuidae) y heliothis ssp. (Lepidóptera: noctuidae), se acentuaron desde los 40 días después de la emergencia (DDE) de las plantas hasta el final de la cosecha. Las plagas que afectaron principalmente los follajes como lyriomiza spp. (Díptera: agromizidae) y aphis spp. Homóptera: aphididae) fueron más constante pero que a partir de los 38 DDE aumentaron sus poblaciones y provocaron un daño severo. El tratamiento que fue protegido desde la emergencia de la planta hasta la cosecha ocupo el primer lugar en fruto/mz, pero sin diferencias significativas entre los periodo de protección a excepción del testigo (sin protección). Así también se colocó en el primer lugar en el número de cajas con frutos exportables por su tamaño/mz, pero sin diferencia significativas con los periodos protegidos desde los 19 y 43 DDE hasta la cosecha. El periodo protegido en ninguna fase de cultivo y el periodo protegido desde la emergencia hasta los 43 DDE ocuparon el primer lugar en fruto no exportable por su tamaño demasiado pequeño. El tratamiento protegido desde los 19 DDE hasta la cosecha presento la mayor ganancia neta. Se presentó deformación de frutos entre 70 y80 %, no presentándose efecto de periodo de protección.

Biología del Chrysopa sp. en la zona algodonera Nicaragua

Este estudio biológico es el primero realizado en Nicaragua, como aporte al programa de Control de Plagas del Algodón. Para detectar la localización geográfica del Chrysopa spp. en Nicaragua se hicieron un total de 180 visitas en 29 municipios, repartidas proporcionalmente según el área sembrada en el municipio algodonero. Se encontró en un 98.4 por ciento de visitas, presencia de Chrysopa spp. Se observo ademas mayor densidad de poblaciones en los meses de enero, febrero y marzo. La obtención del ciclo biológico se obtuvo criando larvas y adultos de este insecto en vasos entomologicos y jaulas recubiertas de cedazo fino respectivamente. El periodo larval fue de 9.75 días, periodo pupal seis días, imagen o adulto 22 a 23 días y huevo 3.5 días. La acción depredadora de las larvas fue verificada suministrando huevos y larvas de Heliothis sp Provewa sp Ephestia sp y Aphis sp. Se encontró que es muy grande la cantidad de insectos depredados y que potencialmente pueden dar un gran beneficio al agricultor.

棉花抗虫基因工程研究

本文通过根农杆菌（Agrobacterium tumfaciens）介导法分别将Signal和KDEL修饰的豇豆胰蛋白酶抑制剂（Cowpea trypsin inhibitor, CpTI）基因、豌豆外源凝集素（Pea lectin, P-Lec）和大豆Kunitz型胰蛋白酶抑制剂（Soybean Kunitz typsin inhibitor, SKTI）双价抗虫基因、雪花莲外源凝集素（Galanthus nivals agglutinin, GNA）基因以及高效复合启动子OM控制的苏云金杆菌（Bacillus thuringiensis, B.t.）杀虫毒蛋白基因导入了陆地棉（Gossypium hirsutum L.）栽培品种新陆早1号、新陆中2号、晋棉7号、冀合321、辽9和晋棉12号，并获得了大批转基因再生植株。 实验中对影响棉花转化和再生的一些条件进行了研究，从根农杆菌培养、棉花无菌苗的制备、转化操作和共培养等方面对转化条件进行了探讨;从激素配化、植物表达载体、外植体类型、基因型等方面对抗性愈伤组织的诱导进行了摸索;从激素、从碳源、培养容器、pH值、抗褐化剂及固化剂的选择等方面对影响植株再生的条件进行了优化。 本文开创性地采用嫁接代替移栽，从而极大地提高了转化植株定植成活率，缩短了缓苗时间并增加了转化植株当代的繁殖系数。 在建立了一套较为高效的陆地棉转化及再生系统基础上，本文还进行了其它转化方式和转化体系的初步探讨。利用棉花幼嫩种子无菌苗下胚轴作为外植体，通过改变愈伤组织诱导培养基配方面提高胚性愈伤组织的诱导频率，进而得到更多的体细胞胚状和再生植株，缩短再生周期;尝试用胚性愈伤组织作为外植体的根农杆菌介导法转化，确定了一些与转化有关的条件;建立了一套棉花茎尖培养程序，为运用基因枪法轰击棉花茎尖分生组织或用根农杆菌直接转化茎尖分生组织，以克服根农杆菌转化棉花时体胚发生的基因型局限开辟了一条新途径。 本文还建立了一种快速鉴定转化植株后代的方法。这一简便方法还有助于进行转基因棉纯合系的筛选以及外源基因的遗传稳定性研究。 转基因植株经Npt-II ELISA、PCR、PCR Southern 检测证明外源抗虫基因CpTI、SKTI、P-lec、GNA以及B.t.基因已存在于转化植株基因组内。修饰的CpTI转基因植株抗棉铃虫（Heliothis armigera Hubner）试验结果表明，其杀虫效果显著优于前期未修饰的CpTI转化植株。P－lec和SKTI双价转基因植株抗棉铃虫试验结果表明，转基因植株对棉铃虫幼虫具有较强的杀虫活力。 目前，已获得转以上抗虫基因棉花T1代植株。为今后进一步将植物基因工程技术应用于棉花遗传改良打下了基础。

Presencia y fluctuación poblacional de artrópodos perjudiciales al cultivo de tomate

p.105-109

Does Gamma-aminobutyric acid function as a plant resistance mechanism against phytophagous insect activity? /

Gamma-aminobutyric acid (GAB A) is a ubiquitous non-protein amino acid synthesized via the decarboxylation of L-glutamate in a reaction catalyzed by the cytosolic enzyme L-glutamate decarboxylase (GAD). In animals it functions as an inhibitory neurotransmitter. In plants it accumulates rapidly in response to various stresses, but its function remains unclear. The hypothesis that GABA accumulation in leaf tissue may function as a plant resistance mechanism against phytophagous insect activity was investigated. GABA accumulation in response to mechanical stimulation, mechanical damage and insect activity was demonstrated. In wt tobacco (Nicotiana tabacum cv Samsun), mechanical stimulation or damage caused GABA to accumulate within 2 min from mean levels of 14 to 37 and 1~9 nmol g-l fresh weight (FW), respectively. In the transgenic tobacco strain CaMVGAD27c overexpressing Petunia GAD, the same treatments caused GABA to accumulate from 12 to 59 and 279 nmol g-l FW, respectively. In the transgenic tobacco strain CaMVGADilC 11 overexpressing Petunia GAD lacking an autoinhibitory domain, mechanical stimulation or damage caused GABA to accumulate from 180 to 309 and 630 nmol g-l FW, respectively. Ambulatory activity by tobacco budworm (TBW) larvae (Heliothis virescens) on leaves of CaMVGAD27c tobacco caused GABA to accumulate from 28 to 80 nmol g-l FW within 5 min. Ambulatory and leaf-rolling activity by oblique banded leaf roller (OBLR) larvae (Choristoneura rosaceana cv Harris) on wt soybean leaves (Glycine max cv Harovinton) caused GABA to accumulate from 60 to 1123 nmol g-l FW within 20 min. Increased GABA levels in leaf tissue were shown to affect phytophagous preference in TBW larvae presented with wt and transgenic tobacco leaves. When presented with leaves of Samsun wt and CaMVGAD27c plants, TBW larvae consumed more wt leaf tissue (640 ± 501 S.D. mm2 ) than transgenic leaf tissue (278 ± 338 S.D. mm2 ) nine times out of ten. When presented with leaves of Samsun wt and CaMVGAD~C11 plants, TBW larvae consumed more transgenic leaf tissue (1219 ± 1009 S.D. mm2 ) than wt leaf tissue (28 ± 31 S.D. mm2 ) ten times out of ten. These results indicate that: (1) ambulatory activity of insect larvae on leaves results in increased GABA levels, (2) transgenic tobacco leaves with increased capacity for GABA synthesis deter feeding, and (3) transgenic tobacco leaves with constitutively higher GABA levels stimulate feeding.

Purification and partial characterization of an aminopeptidase from the midgut tissue of Dysdercus peruvianus

The surface of midgut cells in Hemiptera is ensheathed by a lipoprotein membrane (the perimicrovillar membrane), which delimits a closed compartment with the microvillar membrane, the so-called perimicrovillar space. In Dysdercus peruvianus midgut perimicrovillar space a soluble aminopeptidase maybe involved in the digestion of oligopeptides and proteins ingested in the diet. This D. peruvianus aminopeptidase was purified to homogeneity by ion-exchange chromatography on an Econo-Q column, hydrophobic interaction chromatography on phenyl-agarose column and preparative polyacrylamide gel electrophoresis. The results suggested that there is a single molecular species of aminopeptidase in D. peruvianus midgut. Molecular mass values for the aminopeptidase were estimated to be 106 kDa (gel filtration) and 55 kDa (SDS-PAGE), suggesting that the enzyme occurs as a dimer under native conditions. Kinetic data showed that D. peruvianus aminopeptidase hydrolyzes the synthetic substrates LpNA, RpNA, A beta NA and AsnMCA (K(m)s 0.65, 0.14, 0.68 and 0.74 mM, respectively). The aminopeptidase activity upon LpNA was inhibited by EDTA and 1,10-phenanthroline, indicating the importance of metal ions in enzyme catalysis. One partial sequence of BLAST-identified aminopeptidase was found by random sequencing of the D. peruvianus midgut cDNA library. Semi-quantitative RT-PCR analysis showed that the aminopeptidase genes were expressed throughout the midgut epithelium, in the epithelia of V1, V2 and V3. Malphigian tubules and fat body, but it was not expressed in the salivary glands. These results are important in furthering our understanding of the digestive process in this pest species. (c) 2010 Elsevier Inc. All rights reserved.

Ocorrência de pragas em algodoeiro geneticamente modificado (Bt) e convencional

The study aimed to evaluate the occurrence of pest and management of cotton genetically modified with the introduction of the bacterium Bacillus thunringiensis compared to conventional cotton in the 'Cerrado' region, installing the field experiment in 2007/2008 in Chapadão do Sul-MS. The experimental design was randomized blocks in a 2 × 3 factorial, with two managements of insecticides (with and without insecticides to control lepidopteran) and 3 modes of use of cultivars: 100% transgenic (NUOPAL), 100% non transgenic (Deltaopal) and interior area with transgenic (80% NUOPAL) with non-transgenic border (20% Deltaopal) with five replications. Evaluations were performed weekly of pest infestation, observing in 15 plants per plot the number of Alabama argillacea, Heliothis virescens and Spodoptera frugiperda. Based on these results it was concluded that: the occurrence of A. argillacea and H. virescens was lower in treated transgenic cultivar. There were no differences between transgenic and conventional farming on the occurrence of S. frugiperda.

Ocorrência de pragas e custo de produção em algodoeiro geneticamente modificado (Bt) e convencional

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Interação de proteínas Vip3A e Cry1la10 de Bacillus thuringiensis com atividade inseticida a lepidópteros-praga

Pós-graduação em Agronomia (Genética e Melhoramento de Plantas) - FCAV

Revision of the Neotropical moth genera Mallodeta Butler and Erruca Walker, revalidated (Noctuidae, Arctiinae, Arctiini, Euchromiina)

Mallodeta Butler and Erruca Walker, revalidated, are redescribed and revised. Mallodeta henceforth includes only its type-species, Glaucopis (Lycorea) clavata Walker, and Erruca is resurrected with seven species: E. deyrolii Walker (type-species), E. consors (Walker), new combination, E. erythrarchos (Walker), new combination, E. cardinalis (Hampson), new combination, E. hanga (Herrich-Schaffer), new combination, E. cruenta (Perty), new combination and E. sanguipuncta (Druce), new combination. Six new synonyms are established, four specific and two generic (junior synonyms in parentheses): Zygaena capistrata Fabricius (=Mallodeta cubana Gaede), Glaucopis (Lycorea) clavata Walker (=M. simplex Rothschild), Erruca deyrolii Walker (=Laemocharis aecyra Herrich-Schaffer and Glaucopis (Hyda) sortita Walker), and Erruca Walker (=Aristodaema Wallengren and Rezia Kirby). Lectotypes are designated to the following species: Erruca deyrolii Walker, Laemocharis deyrollei Herrich-Schaffer, Laemocharis hanga Herrich-Schaffer, Laemocharis aecyra Herrich-Schaffer, Laemocharis norma Herrich-Schaffer, Cosmosoma cardinalis Hampson and Mallodeta sanguipuncta Druce. Illustrations of adults and male and female genitalia of Mallodeta and Erruca are provided, as well as a key to the species of the latter.

Characterization of a Toxoneuron nigriceps (Viereck) (Hymenoptera: Braconidae) - derived chitinase and its potential for pest control

The larval endoparasitoid Toxoneuron nigriceps (Viereck) (Hymenoptera: Braconidae) has a toolbox of biological weapons to secure for host colonization and the successful parasitization of its host Heliothis virescens (F.) (Lepidoptera: Noctuidae). The cDNA of a putative chitinase has been previously isolated and initially characterized from teratocytes of this parasitoid among the plethora of molecules available in the venom and calyx fluids injected by females, oral and/or anal secretions released by the parasitoid larvae and/or produced by the expression of genes of the symbiotic associated polydnavirus. This putative chitinase has been initially associated with the host cuticle digestion to allow for parasitoid egression and with the asepsis of the host environment, acting as an antimicrobial. As chitinases are commonly expressed in plants against plant pathogens, the chitinase derived from the teratocytes of T. nigriceps is a potential tool for the development of insect pest control methods based on the disruption of the perithrophic membrane of herbivores. Therefore, we aimed to characterize the activity of the putative chitinase from teratocytes of T. nigriceps (Tnchi) produced using the Escherichia coli expression system and its potential to control H. virescens larvae when expressed into transgenic tobacco plants. The purified E. coli-produced Tnchi protein showed no chitinolitic activity, but was active in binding with colloidal and crystalline chitins in water and with colloidal chitin in buffered solution (pH = 6.74). Transgenic tobacco plants showed no enhanced chitinolitic activity relative to control plants, but survival of three-day old larvae of H. virescens was severely affected when directly fed on transgenic tobacco leaves expressing the recombinant Tnchi protein. Some properties of the Tnchi protein and the potential use of Tnchi-transgenic plants to control plant pests are discussed. (c) 2012 Elsevier Inc. All rights reserved.

Assessment of the high-dose concept and level of control provided by MON 87701 x MON 89788 soybean against Anticarsia gemmatalis and Pseudoplusia includens (Lepidoptera: Noctuidae) in Brazil

BACKGROUND: Genetically modified MON 87701 X MON 89788 soybean (Glycine max), which expresses the Cry1Ac and EPSP-synthase proteins, has been registered for commercial use in Brazil. To develop an Insect Resistance Management (IRM) program for this event, laboratory and field studies were conducted to assess the high-dose concept and level of control it provides against Anticarsia gemmatalis and Pseudoplusia includens. RESULTS: The purified Cry1Ac protein was more active against A. gemmatalis [LC50 (FL 95%) = 0.23 (0.150.34) mu g Cry1Ac mL-1 diet] than P. includens [LC50 (FL 95%) = 3.72 (2.654.86) mu g Cry1Ac mL-1 diet]. In bioassays with freeze-dried MON 87701X MON 89788 soybean tissue diluted 25 times in an artificial diet, there was 100% mortality of A. gemmatalis and up to 95.79% mortality for P. includens. In leaf-disc bioassays and under conditions of high artificial infestation in the greenhouse and natural infestation in the field, MON 87701X MON 89788 soybean showed a high level of efficacy against both target pests. CONCLUSIONS: The MON 87701X MON 89788 soybean provides a high level of control against A. gemmatalis and P. includes, but a high-dose event only to A. gemmatalis. Copyright (c) 2012 Society of Chemical Industry