Antifeedant Effect of Some Medicinal Plant Extracts Against Rice Weevil.

BACKGROUND AND OBJECTIVE: At present, insecticides with raw materials of active compounds from plants have become a strategic alternative, because they are easily obtained and produced. In an attempt to find natural and cheaper methods for the control of rice weevil (Sitophilus oryzae L.), some useless waste of local medicinal plant extracts was evaluated. This study aimed to evaluate the antifeedant effect of various medicinal plant extracts on rice weevil (Sitophilus oryzae L.). MATERIALS AND METHODS: The experimental study using a completely randomized design of two factors, i.e., source of the extract in 7 types of medicinal plant waste and the exposure time of extracts in 0, 1, 2 and 3 h. The antifeedant effects were evaluated after 3 weeks by measuring feed consumption (FC), feeding deterrent index (FDI) and rice weevils' mortality. Data were analyzed using variance analysis. RESULTS: The statistical analysis showed that the differences in exposure time contributed significant influence (α<0.05) on FC, FDI and mortality. The lowest FC occurred at the 3 h exposure treatment, i.e. on the extract of Amomum cardamom plant, while the highest FC was found in the treatment of Curcuma mangga plant extract. The values of FDI and mortality at the 3 h extract exposure were 31.53-87.16% and 42.4-64%, respectively with the highest value in the treatment of A. cardamomum. CONCLUSION: At last, these findings inform peoples that the waste of medicinal plants, especially cardamom is possible to be developed as botanical insecticides for rice weevils.

Evolutionarily conserved plant genes responsive to root-knot nematodes identified by comparative genomics.

Root-knot nematodes (RKNs, genus Meloidogyne) affect a large number of crops causing severe yield losses worldwide, more specifically in tropical and sub-tropical regions. Several plant species display high resistance levels to Meloidogyne, but a general view of the plant immune molecular responses underlying resistance to RKNs is still lacking. Combining comparative genomics with differential gene expression analysis may allow the identification of widely conserved plant genes involved in RKN resistance. To identify genes that are evolutionary conserved across plant species, we used OrthoFinder to compared the predicted proteome of 22 plant species, including important crops, spanning 214 Myr of plant evolution. Overall, we identified 35,238 protein orthogroups, of which 6,132 were evolutionarily conserved and universal to all the 22 plant species (PLAnts Common Orthogroups-PLACO). To identify host genes responsive to RKN infection, we analyzed the RNA-seq transcriptome data from RKN-resistant genotypes of a peanut wild relative (Arachis stenosperma), coffee (Coffea arabica L.), soybean (Glycine max L.), and African rice (Oryza glaberrima Steud.) challenged by Meloidogyne spp. using EdgeR and DESeq tools, and we found 2,597 (O. glaberrima), 743 (C. arabica), 665 (A. stenosperma), and 653 (G. max) differentially expressed genes (DEGs) during the resistance response to the nematode. DEGs' classification into the previously characterized 35,238 protein orthogroups allowed identifying 17 orthogroups containing at least one DEG of each resistant Arachis, coffee, soybean, and rice genotype analyzed. Orthogroups contain 364 DEGs related to signaling, secondary metabolite production, cell wall-related functions, peptide transport, transcription regulation, and plant defense, thus revealing evolutionarily conserved RKN-responsive genes. Interestingly, the 17 DEGs-containing orthogroups (belonging to the PLACO) were also universal to the 22 plant species studied, suggesting that these core genes may be involved in ancestrally conserved immune responses triggered by RKN infection. The comparative genomic approach that we used here represents a promising predictive tool for the identification of other core plant defense-related genes of broad interest that are involved in different plant-pathogen interactions.

Protection of rice against Nilaparvata lugens by direct toxicity of sodium selenate.

Nilaparvata lugens is one of the most notorious pest insects of cultured rice, and outbreaks of N. lugens cause high economic losses each year. While pest control by chemical pesticides is still the standard procedure for treating N. lugens infections, excessive use of these insecticides has led to the emergence of resistant strains and high pesticide residues in plants for human consumption and the environment. Therefore, novel and environment-friendly pest control strategies are needed. In previous studies, selenium was shown to protect selenium-accumulating plants from biotic stress. However, studies on nonaccumulator (crop) plants are lacking. In this study, rice plants (Oryza sativa, Nipponbare) were treated with sodium selenate by seed priming and foliar spray and then infested with N. lugens. Brown planthoppers feeding on these plants showed increased mortality compared to those feeding on control plants. Treatment of the plants with sodium selenate did not affect the enzymes involved in the biosynthesis of the plant stress hormones jasmonic acid and salicylic acid, suggesting that the observed insect mortality cannot be attributed to the activation of these hormonal plant defenses. Feeding assays using an artificial diet supplemented with sodium selenate revealed direct toxicity toward N. lugens. With a low concentration of 6.5 ± 1.5 µM sodium selenate, half of the insects were killed after 3 days. In summary, sodium selenate treatment of plants can be used as a potential alternative pest management strategy to protect rice against N. lugens infestation through direct toxicity.

Risk versus reward: host dependent parasite mortality rates and phenotypes in the facultative generalist Triphysaria versicolor.

BACKGROUND: Parasitic plants engage in a complex molecular dialog with potential host plants to identify a host and overcome host defenses to initiate development of the parasitic feeding organ, the haustorium, invade host tissues, and withdraw water and nutrients. While one of two critical signaling events in the parasitic plant life cycle (germination via stimulant chemicals) has been relatively well-studied, the signaling event that triggers haustorium formation remains elusive. Elucidation of this poorly understood molecular dialogue will shed light on plant-plant communication, parasitic plant physiology, and the evolution of parasitism in plants. RESULTS: Here we present an experimental framework that develops easily quantifiable contrasts for the facultative generalist parasitic plant, Triphysaria, as it feeds across a broad range of diverse flowering plants. The contrasts, including variable parasite growth form and mortality when grown with different hosts, suggest a dynamic and host-dependent molecular dialogue between the parasite and host. Finally, by comparing transcriptome datasets from attached versus unattached parasites we gain insight into some of the physiological processes that are altered during parasitic behavior including shifts in photosynthesis-related and stress response genes. CONCLUSIONS: This work sheds light on Triphysaria's parasitic life habit and is an important step towards understanding the mechanisms of haustorium initiation factor perception, a unique form of plant-plant communication.

Eugenol diffusion coefficient and its potential to control Sitophilus zeamais in rice.

Given the insecticidal potential of eugenol as a fumigant, this work aimed to determine the diffusion coefficient of eugenol emanating from a pure standard solution (99%), as well as from clove essential oil (Eugenia caryophillata Thunb. (Myrtaceae)) through rice grain; to chemically analyse the volatile composition of commercially available eugenol and clove essential oil; and to evaluate the mortality of Sitophilus zeamais Motschulsky (Coleoptera: curculionidae) after exposure to eugenol inside a test chamber filled with rice. The solid phase microextraction method of extracting and quantifying eugenol by gas chromatography presented a good analytical response for the quantification of the analyte. There was no significant difference between the diffusion coefficient of eugenol diffusing from pure eugenol or from clove essential oil. The diffusion coefficient of eugenol through rice with the conditions herein adopted is 1.09 × 10-3 cm2 s-1. The characterization of clove essential oil confirmed the presence of eugenol as its major component (74.25%). A difference was observed in the composition of the distinct phases evaluated. The exposure of adult S. zeamais to diffused eugenol from pure eugenol over seven days resulted in significantly higher mortality rates (~37%) than eugenol diffused from clove essential oil (~11%). No differences in mortality rates were observed in individuals placed at different positions inside the test chamber during eugenol fumigation.

Toxicological effect of underutilized plant, Cleistanthus collinus leaf extracts against two major stored grain pests, the rice weevil, Sitophilus oryzae and red flour beetle, Tribolium castaneum.

Toxicity and repellency activity of Cleistanthus collinus (Roxb.) (CC) leaf extracts were evaluated against rice weevil, Sitophilus oryzae (L.) and red flour beetle, Tribolium castaneum (Herbst) under laboratory condition. Five concentration(s) (1%, 1.5%, 2%, 2.5% and 4%) with two controls (acetone and water) treatments along with deltamethrin were used for direct and contact residual toxicity. The insect that survived after CC exposure were transferred to an untreated feeding substrate and the population buildup of subsequent two generation were recorded after 30 (F1) and 60 days (F2). In the contact residual toxicity, highest CC concentration (4%) produced 75% mortality in S. oryzae and 62.5% mortality in T. castaneum during 7 days of exposure, whereas in direct toxicity the mortality were 81% and 58% respectively, for S. oryzae and T. castaneum. The long term effect of CC was apparent in both the insect species, where F2 populations were significantly decreased in the CC treatments. CC treatment at 4% produced similar adult mortality in comparison to deltamethrin at 1%. In addition, repellent activity of CC extracts was observed against both S. oryzae and T. castaneum. This is the first step towards assessing the scientific basis for the understanding the effectiveness of CC extracts against stored grain pests and it could be a viable eco-friendly option for stored grain insect pest management.

Protection of Grain Products from Sitophilus oryzae (L.) Contamination by Anti-Insect Pest Repellent Sachet Containing Allyl Mercaptan Microcapsule.

The purpose of this study was to develop an anti-insect pest repellent sachet to prevent Sitophilus oryzae (L.) (Coleoptera: Curculionidae) contamination in grain packaging. The anti-insect pest activities of essential oils (EOs) from garlic (Allium Sativum), ginger (Zingiber Officinalis), black pepper (Piper nigrum), onion (Allium cepa), and fennel (Foeniculum vulgare) as well as major compounds (allyl disulfide, AD; allyl mercaptan, AM) isolated from of garlic and onion (AD and AM) were measured against S. oryzae. The results revealed that garlic EO, onion EO, AD, and AM showed strong fumigant insecticidal activities. Among these, AM showed the highest acetylcholinesterase (AChE) inhibition rate, indicating that the fumigation insecticidal efficacy of AM is related with its AChE inhibition ability. Subsequently, the microcapsules were produced with a high efficiency (80.02%) by using AM as a core material and rice flour as a wall material. Finally, sachet composed of rice flour microcapsule containing 2% AM (RAM) was produced. Repellent assay was performed to measure anti-insect pest ability of the RAM sachet, showed remarkable repelling effect within 48 h both in the presence or absence of attractant. In a release profile of RAM sachet, it was expected to last over 20 mo during the distribution period of brown rice. Moreover, RAM sachet showed no undesirable changes to the sensory properties of the rice both before and after cooking. Taken together, these results suggest that the newly developed RAM sachet could be used as a packaging material to protect grain products from S. oryzae contamination. PRACTICAL APPLICATION: The rice weevil, Sitophilus oryzae (L.) (Coleoptera: Curculionidae), causes damages to stored products and its contamination in grain products has become a major problem in cereal market. To preserve brown rice, an anti-insect pest repellent sachet containing 2% allyl mercaptan was newly developed and it showed remarkable repellent abilities against S. oryzae. It could be used as an active food packaging system to protect grain products from insect pest contamination.

Higher Fertilizer Inputs Increase Fitness Traits of Brown Planthopper in Rice.

Rice (Oryza sativa L.) is the primary staple food source for more than half of the world's population. In many developing countries, increased use of fertilizers is a response to increase demand for rice. In this study, we investigated the effects of three principal fertilizer components (nitrogen, phosphorus and potassium) on the development of potted rice plants and their effects on fitness traits of the brown planthopper (BPH) [Nilaparvata lugens (Stål) (Homoptera: Delphacidae)], which is a major pest of rice in Bangladesh and elsewhere. Compared to low fertilizer inputs, high fertilizer treatments induced plant growth but also favored BPH development. The BPH had higher survival, developed faster, and the intrinsic rate of natural increase (r m ) was higher on well-fertilized than under-fertilized plants. Among the fertilizer inputs, nitrogen had the strongest effect on the fitness traits of BPH. Furthermore, both the "Plant vigor hypothesis" and the "Plant stress hypothesis" were supported by the results, the former hypothesis more so than the latter. These hypotheses suggest that the most suitable/attractive hosts for insect herbivores are the most vigorous plants. Our findings emphasized that an exclusive focus on yield increases through only enhanced crop fertilization may have unforeseen, indirect, effects on crop susceptibility to pests, such as BPH.

How the rice weevil breaks down the pectin network: Enzymatic synergism and sub-functionalization.

Pectin is the most complex polysaccharide in nature and highly abundant in plant cell walls and middle lamellae, where it functions in plant growth and development. Phytopathogens utilize plant pectin as an energy source through enzyme-mediated degradation. These pectolytic enzymes include polygalacturonases (PGs) of the GH28 family and pectin methylesterases (PMEs) of the CE8 family. Recently, PGs were also identified in herbivorous insects of the distantly related plant bug, stick insect and Phytophaga beetle lineages. Unlike all other insects, weevils possess PMEs in addition to PGs. To investigate pectin digestion in insects and the role of PMEs in weevils, all PME and PG family members of the rice weevil Sitophilus oryzae were heterologously expressed and functionally characterized. Enzymatically active and inactive PG and PME family members were identified. The loss of activity can be explained by a lack of substrate binding correlating with substitutions of functionally important amino acid residues. We found subfunctionalization in both enzyme families, supported by expression pattern and substrate specificities as well as evidence for synergistic pectin breakdown. Our data suggest that the rice weevil might be able to use pectin as an energy source, and illustrates the potential of both PG and PME enzyme families to functionally diversify after horizontal gene transfer.

Phloem-specific expression of the lectin gene from Allium sativum confers resistance to the sap-sucker Nilaparvata lugens.

Rice production is severely hampered by insect pests. Garlic lectin gene (ASAL) holds great promise in conferring protection against chewing (lepidopteran) and sap-sucking (homopteran) insect pests. We have developed transgenic rice lines resistant to sap-sucking brown hopper (Nilaparvata lugens) by ectopic expression of ASAL in their phloem tissues. Molecular analyses of T0 lines confirmed stable integration of transgene. T1 lines (NP 1-2, 4-3, 11-6 & 17-7) showed active transcription and translation of ASAL transgene. ELISA revealed ASAL expression was as high as 0.95% of total soluble protein. Insect bioassays on T2 homozygous lines (NP 18 & 32) revealed significant reduction (~74-83%) in survival rate, development and fecundity of brown hoppers in comparison to wild type. Transgenics exhibited enhanced resistance (1-2 score) against brown hoppers, minimal plant damage and no growth penalty or phenotypic abnormalities.

BIOEFFICACY OF PIPER RIBESIOIDES (PIPERACEAE) EXTRACTS AGAINST NILAPARVATA LUGENS STAL. (HOMOPTERA: DELPHACIDAE).

The extracts of stem and seeds of Piper ribesioides (Piperaceae) were prepared in hexane, dichloromethane, ethyl acetate and ethanol and evaluated for their efficacy against 3-day-old adult brown plant hoppers, Nilaparvata lugens Stal under laboratory conditions. The treatment was given topically and results suggest the dose dependent response related to exposure time. The ethyl acetate extract was the best with an LC50 = 738 ppm-, 24 h post-treatment. There was substantial increase in activity (LC50 = 27.298 ppm) with the increase in duration of exposure. This trend was observed in all treatments that suggests the potential of Piper ribesioides as an insecticide for N. lugens as an alternative control.

Molecular characterization of a ryanodine receptor gene in the rice leaffolder, Cnaphalocrocis medinalis (Guenée).

Ryanodine receptors (RyRs) are the targets of two novel classes of synthetic insecticidal chemicals, phthalic acid diamides and anthranilic diamides. Isolation of full-length RyR cDNAs is a critical step towards the structural and functional characterization of insect RyRs and an understanding of the molecular mechanisms underlying the species selective toxicity of diamide insecticides. However, there has been little research on the insect RyR genes due to the high molecular weight of the RyR proteins. In this study, we isolated a full-length RyR cDNA (named as CmRyR) from Cnaphalocrocis medinalis, an important rice pest throughout Southeast Asia. The composite CmRyR gene contains an ORF of 15264 bp encoding a protein of 5087 amino acid residues, which shares 79% overall identity with its Drosophila melanogaster homologue. All hallmarks of the RyR proteins are conserved in the CmRyR protein, suggesting that CmRyR is a structural and functional analogue of known RyRs. A multiple sequence alignment illustrates that the insect RyRs share high levels of amino acid sequence identity at the the COOH-terminal region. However, the amino acid residues analogous to the CmRyR residues N(4922), N(4924), N(4935), L(4950), L(4981), N(5013) and T(5064) are unique to lepidopteran RyRs compared with non-lepidopteran insect RyRs. This finding suggests that these residues may be involved in the differences in channel properties between lepidopteran and non-lepidopteran insect RyRs and in the species selective toxicity of diamide insecticides. Furthermore, two alternative splicing sites were identified in the CmRyR gene, one of which was located in the central part of the predicted second SPRY domain. Diagnostic PCR showed that the inclusion frequencies of two mutually exclusive exons (a/b) and one optional exon (c) differed between developmental stages or adult anatomical regions. Our results imply that alternative splicing may be a major means of generating functional diversity in C. medinalis RyR channel.

Effect of arbuscular mycorrhizal fungi (Glomus intraradices) on the oviposition of rice water weevil (Lissorhoptrus oryzophilus).

Root-feeding insects are important drivers in ecosystems, and links between aboveground oviposition preference and belowground larval performance have been suggested. The root-colonizing arbuscular mycorrhizal fungi (AMF) play a central role in plant nutrition and are known to change host quality for root-feeding insects. However, it is not known if and how AMF affect the aboveground oviposition of insects whose offspring feed on roots. According to the preference-performance hypothesis, insect herbivores oviposit on plants that will maximize offspring performance. In a greenhouse experiment with rice (Oryza sativa), we investigated the effects of AMF (Glomus intraradices) on aboveground oviposition of rice water weevil (Lissorhoptrus oryzophilus), the larvae of which feed belowground on the roots. Oviposition (i.e., the numbers of eggs laid by weevil females in leaf sheaths) was enhanced when the plants were colonized by AMF. However, the leaf area consumed by adult weevils was not affected. Although AMF reduced plant biomass, it increased nitrogen (N) and phosphorus concentrations in leaves and N in roots. The results suggest that rice water weevil females are able to discriminate plants for oviposition depending on their mycorrhizal status. The discrimination is probably related to AMF-mediated changes in plant quality, i.e., the females choose to oviposit more on plants with higher nutrient concentrations to potentially optimize offspring performance. AMF-mediated change in plant host choice for chewing insect oviposition is a novel aspect of below- and aboveground interactions.

Efficacy of layer treatment with methoprene for control of Rhyzopertha dominica (Coleoptera: Bostrychidae) on wheat, rice and maize.

BACKGROUND: Insect growth regulators are promising alternatives to traditional pesticides in stored grain. The efficacy of the juvenile hormone analogue methoprene was evaluated as a layer treatment in a laboratory experiment for control of Rhyzopertha dominica (F.) (Coleoptera: Bostrychidae) in wheat, rice and maize. RESULTS: Adults of R. dominica were placed in vials containing 33, 26 and 29 g (to a depth of 6.5 cm) of wheat, rice and maize, respectively, that was entirely or partially treated with 1, 5 or 10 mg kg(-1) methoprene. In wheat and rice, the layer treatments were not as effective as the whole-grain treatment, but there was decreased progeny production as the application rate increased. However, on maize the partial treatments were as effective as the whole-grain treatment at 5 and 10 mg kg(-1) . CONCLUSIONS: The results suggest that partial layer treatments with methoprene can be used to control R. dominica on maize but may not be effective for control of this species on wheat and rice.

Pyramided rice lines harbouring Allium sativum (asal) and Galanthus nivalis (gna) lectin genes impart enhanced resistance against major sap-sucking pests.

We have developed transgene pyramided rice lines, endowed with enhanced resistance to major sap-sucking insects, through sexual crosses made between two stable transgenic rice lines containing Allium sativum (asal) and Galanthus nivalis (gna) lectin genes. Presence and expression of asal and gna genes in pyramided lines were confirmed by PCR and western blot analyses. Segregation analysis of F2 progenies disclosed digenic (9:3:3:1) inheritance of the transgenes. Homozygous F3 plants carrying asal and gna genes were identified employing genetic and molecular methods besides insect bioassays. Pyramided lines, infested with brown planthopper (BPH), green leafhopper (GLH) and whitebacked planthopper (WBPH), proved more effective in reducing insect survival, fecundity, feeding ability besides delayed development of insects as compared to the parental transgenics. Under infested conditions, pyramided lines were found superior to the parental transgenics in their seed yield potential. This study represents first report on pyramiding of two lectin genes into rice exhibiting enhanced resistance against major sucking pests. The pyramided lines appear promising and might serve as a novel genetic resource in rice breeding aimed at durable and broad based resistance against hoppers.

Backbone cyclised peptides from plants show molluscicidal activity against the rice pest Pomacea canaliculata (golden apple snail).

Golden apple snails ( Pomacea canaliculata) are serious pests of rice in South East Asia. Cyclotides are backbone cyclized peptides produced by plants from Rubiaceae and Violaceae. In this study, we investigated the molluscicidal activity of cyclotides against golden apple snails. Crude cyclotide extracts from both Oldenlandia affinis and Viola odorata plants showed molluscicidal activity comparable to the synthetic molluscicide metaldehyde. Individual cyclotides from each extract demonstrated a range of molluscicidal activities. The cyclotides cycloviolacin O1, kalata B1, and kalata B2 were more toxic to golden apple snails than metaldehyde, while kalata B7 and kalata B8 did not cause significant mortality. The toxicity of the cyclotide kalata B2 on a nontarget species, the Nile tilapia ( Oreochromis niloticus), was three times lower than the common piscicide rotenone. Our findings suggest that the existing diversity of cyclotides in plants could be used to develop natural molluscicides.

Effect of biopesticides on the lactate dehydrogenase (LDH) of the rice leaffolder, Cnaphalocrocis medinalis (Guenée) (Insecta: Lepidoptera: Pyralidae).

The effects of bacterial toxins (Bacillus thuringiensis) and botanical insecticides (Azadirachta indica and Vitex negundo) on lactate dehydrogenase (LDH) activity in Cnaphalocrocis medinalis (Guenée) (the rice leaffolder) were evaluated. Bacterial toxins and botanical insecticides affected the LDH activity individually and in combination. When they were combined, the effect was more severe at low concentration. There was a decrease in enzyme activity over controls at all concentrations tested. The combined effect of the three biopesticides resulted in a considerable decrease in enzyme activity, indicating strong enzyme inhibition. Clear dose-response relationships were established with respect to enzyme activity.

Insecticidal spider venom toxin fused to snowdrop lectin is toxic to the peach-potato aphid, Myzus persicae (Hemiptera: Aphididae) and the rice brown planthopper, Nilaparvata lugens (Hemiptera: Delphacidae).

The SFI1/GNA fusion protein, comprising of snowdrop lectin (Galanthus nivalis agglutinin, GNA) fused to an insecticidal spider venom neurotoxin (Segestria florentina toxin 1, SFI1) was tested for toxicity against the rice brown planthopper Nilaparvata lugens (Stål) and the peach-potato aphid Myzus persicae (Sulzer) by incorporation into artificial diets. Significant effects on the mortality of N. lugens were observed, with 100% of the insects fed on the SFI1/GNA fusion protein diet dead by day 7. The survival of the aphid M. persicae was also reduced when fed on the SFI1/GNA fusion protein. After 14 days, only 49% of the aphids that were fed on the fusion protein were still alive compared with approximately 90% of the aphids fed on the control diet or on diet containing GNA only. The SFI1/GNA fusion protein also slowed the development of M. persicae, and the reproductive capacity of the aphids fed on the SFI1/GNA fusion protein was severely reduced. The ability of GNA to act as a carrier protein, and deliver the SFI1 neurotoxin to the haemolymph of N. lugens, following oral ingestion, was investigated. The successful delivery of intact SFI1/GNA fusion protein to the haemolymph of these insects was shown by western blotting. Haemolymph taken from the insects that were fed on the fusion protein contained two GNA-immunoreactive proteins of molecular weights corresponding to GNA and to the SFI1/GNA fusion protein.

The novel botanical insecticide for the control brown planthopper (Nilaparvata lugens Stal.).

Brown planthopper, Nilaparvata lugens Stal., (BPH) was the most devastating insect pest on rice in many partS of Asia. The Outbreak of BPH? which is resistant to many synthetic insecticides can cause total rice crop loss. This research was done to evaluate the efficiency of mangostin from the pericarp of mangosteen fruit extract (Garcina mangostana L.) as the alternative control of BPH. The pericarp of mangosteen fruit was extracted by Soxhlet apparatus using ethanol as a solvent and purified by chromatography method then qualified structure by 2D-NMR, MS and IR. The crude extracts contained mangostin ca. 2.956% w/w. This extract was trailed by the topical sprayer method with 1st, 2nd, 3rd, 4th and 5th nymph and adult BPH shows toxicity in term of LC50 ca. 1.39, 2.26, 5.44, 4.49, 4.03 and 3.84 % w/v at 24 h exposure, respectively. The in vitro enzyme activity from BPH survived after 24 h exposure and showed to inhibit the carboxylesterase (CarE), acetylchoinesterase (AchE) and glutathione-S-transferase (GST) activities which the correction factors of CarE, AchE and GST indicated ca. 1.21-2.05 fold, 1.24-2.50 fold and 1.01-3.34 fold, respectively. Moreover, the data shows that the carboxylesterase may play an important role to detoxify this extract. The results suggested that pericarp of mangosteen fruit extract which have mangostin as active ingredient compound shows mechanism as the inhibitor of detoxification enzymes. Thus, it is likely to be uses this extract as an insecticide alternative to the control of BPH.

Insect resistance to Nilaparvata lugens and Cnaphalocrocis medinalis in transgenic indica rice and the inheritance of gna+sbti transgenes.

Molecular genetic analysis and insect bioassay of transgenic indica rice 'Zhuxian B' plants carrying snowdrop lectin gene (gna) and soybean trypsin inhibitor gene (sbti) were investigated in detail. PCR, 'dot' blot and PCR-Southern blot analysis showed that both transgenes had been incorporated into the rice genome and transmitted up to R3 progeny in most lines tested. Some transgenic lines exhibited Mendelian segregation, but the other showed either 1:1 (positive: negative for the transgenes) or other aberrant segregation patterns. The segregation patterns of gna gene crossed between R2 and R3 progeny. In half of transgenic R3 lines, gna and sbti transgenes co-segregated. Two independent homozygous lines expressing double transgenes were identified in R3 progeny. Southern blot analysis demonstrated that the copy numbers of integrated gna and sbti transgenes varied from one to ten in different lines. Insect bioassay data showed that most transgenic plants had better resistance to both Nilaparvata lugens (Stahl) and Cnaphalocrocis medinalis (Guenee) than wild-type plants. The insect resistance of transgenic lines increased with the increase in transgene positive ratio in most of the transgenic lines. In all, we obtained nine lines of R3 transgenic plants, including one pure line, which had better resistance to both N lugens and C medinalis than wild-type plants.