Evaluation of Lecanicillium longisporum, Vertalec against the cotton aphid, Aphis gossypii, and cucumber powdery mildew, Sphaerotheca fuliginea in a greenhouse environment

The commercial mycopesticide, Vertalec^® based on Lecanicillium longisporum, was evaluated for simultaneous control of aphids and powdery mildew on cucumbers in a greenhouse setting where temperature and RH were allowed to fluctuate within normal operating ranges. Five to six week old cucumber plants were inoculated with either Sphaerotheca fuliginea (Sf) spores, cotton aphids (Aphis gossypii) or both. Vertalec, Vertalec containing irradiation-inactivated blastospores (II Vertalec) or sterilized water (control) were applied to the plants 1, 4, and 7 days later. Vertalec treatment provided complete control of aphids 16 days after aphid inoculation, whereas effects of the II Vetalec were not significantly different from the water-treated control. The number of powdery mildew spots on cucumber leaves and the number of S. fuliginea spores in each spot were significantly lower in Vertalec-treated plants than II Vertalec-treated plants or the controls, whereas numbers in the II Vertalec treatment were lower that the untreated control. These results demonstrate that Vertalec has potential for simultaneous management of both cotton aphid and powdery mildew in greenhouse cucumber production.     

Endophytic Lecanicillium lecanii and Beauveria bassiana reduce the survival and fecundity of Aphis gossypii following contact with conidia and secondary metabolites

Lecanicillium lecanii and Beauveria bassiana are important entomopathogens of Aphis gossypii. Their capacity to colonize crop plants is also becoming widely recognized. Their presence in crop plants indicates the possibility of a much greater potential for contact between insect and fungus than previously recognized. The present experiment aimed to study the effects of endophytic strains of these fungi on the survival, and reproduction of A. gossypii. Contact with conidia of both fungi significantly reduced the rate and period of reproduction of A. gossypii. The culture filtrates of L. lecanii and B. bassiana significantly increased mortality and feeding-choice experiments indicate that insects may be able to detect metabolites of the fungi. The culture filtrate of L. lecanii also significantly reduced the reproduction of the aphid. The ethyl acetate and methanolic fractions of the culture filtrate and of mycelia of L. lecanii also caused significant mortality and reduced fecundity of A. gossypii. The methanolic fractions of mycelia of B. bassiana caused significant mortality of A. gossypii. The present investigations indicated that A. gossypii is affected by contact with both conidia and fungal metabolites. This broad influence indicates that these fungi may have a role in regulating insect pest populations.     

Insecticidal effects of Ungernia severtzovii bulb extracts against the grain aphid Schizaphis graminum (Rondani)

Insecticidal effect of Ungernia severtzovii bulbs extract has been investigated against the grain aphid Schizaphis graminum. LC₅₀ value of the ethanolic extract of U. severtzovii against the female aphids was 0.235% (2.35 g/l). There was 100% mortality of the subsequent nymphs of the treated females at 0.25-1.0%. The extract was equally active against the nymphs at the highest concentrations. Hexane and ethanolic extracts were the most active against the aphids of all the extracts. Fractionation of the hexane extract by TLC yielded three fractions. Fraction II was the most active (>90% mortality) against the aphids. Xanthoxylin was identified as the major constituent in fraction II of the hexane extract and may be responsible for the insecticidal effect of U. severtzovii.     

Identification of an insecticidal polyacetylene derivative from Chrysanthemum macrotum leaves

Compounds responsible for insecticidal properties of Chrysanthemum macrotum (D.R.) Ball. leaves against Spodoptera littoralis Boiduval caterpillars have been investigated. The screening of the insecticidal activity was performed by incorporating methanol, buthanol or ethyl acetate extracts, or some chromatographic fractions to the caterpillars’ artificial diet. It was noted that extracts and fractions ameliorated or disturbed nutritional indexes, being not always toxic for caterpillars. Among the tested fractions, one pure compound with a high insecticidal activity (percentage of mortality 66.7%) was purified. The nuclear magnetic resonance study allowed its identification as a polyacetylene derivative, in particular a spiroketal enol ether one.     

Effects of crude seed extracts of Annona atemoya and Annona squamosa L. against the cabbage looper, Trichoplusia ni in the laboratory and greenhouse

Antifeedant, growth inhibitory and toxic effects of crude seed extracts of Annona squamosa and Annona atemoya from Fazenda Viveiro Bona, Parasisópolis – Minas Gerais, Brazil, were evaluated against the cabbage looper, Trichoplusia ni (Hübner) (Lepidoptera: Noctuidae) using different bioassays. Crude methanolic seed extracts deterred feeding of third instar T. ni larvae in a leaf disc choice bioassay. A. squamosa was ∼10 times more active as a feeding deterrent than A. atemoya (DC₅₀ = 2.3 mg/ml vs. 20.1 mg/ml). A. squamosa was ∼three times more active as a growth inhibitor than A. atemoya (EC₅₀ = 38.0 ppm vs. 117.0 ppm). Methanolic seed extracts of A. squamosa and A. atemoya were toxic to third instar T. ni larvae both through topical and oral application. A. squamosa was more toxic through feeding (LC₅₀ = 167.5 ppm vs. 382.4 ppm) whereas, A. atemoya exerted greater toxicity via topical application (LC₅₀ = 301.3 μg/larva vs. 197.7 μg/larva). Both A. squamosa and A. atemoya extracts reduced leaf area consumption and larval growth in a greenhouse experiment. Our results indicate that both A. squamosa and A. atemoya have potential for development as botanical insecticides, especially for local use in Brazil.     

Hybanthus parviflorus (Violaceae): Insecticidal activity of a South American plant

Hybanthus parviflorus is a perennial shrub, widely distributed in the tropical and subtropical regions of America, and is known as ‘violetilla’ in Argentina. Previous phytochemical studies of this species led us to the isolation and determination of the primary structure of a novel macrocyclic polypeptide, the cyclotide hypa A. Here the insecticidal activities of extracts of H. parviflorus were determined against Ceratitis capitata Wied., the Mediterranean fruit fly or ‘Medfly’. Mortality in the different life stages of Medfly, total mortality and modifications of the insect’s physiology caused by 50% EtOH and CH₂Cl₂ extracts of H. parviflorus were evaluated. In addition, we determined the occurrence of ursolic acid, β-sitosterol and the polyphenols quercetin, quercetin-3-methyl ether, apigenin, luteolin, kaempferol, rutin, caffeic acid and chlorogenic acid. The promising insecticidal activity of 50% EtOH extracts of H. parviflorus and its purified fractions was related to the presence of cyclotides. The insecticidal activity of CH₂Cl₂ extracts could be related to the presence of polyphenols, ursolic acid and β-sitosterol.     

Comparative assessment of antioxidant and cholinesterase inhibitory properties of the marigold extracts from Calendula arvensis L. and Calendula officinalis L.

Calendula sp. is an important medicinal and industrial plant with various bioactivities. In this study, we examined enzyme inhibitory effects of the n-hexane, dichloromethane, acetone, ethyl acetate, methanol, and water extracts of the leaf and flowers of Calendula arvensis L. and C. officinalis L. against acetylcholinesterase (AChE) and butyrylcholinesterase (BChE). The extracts were screened for their antioxidant activity using 2,2-diphenyl-1-picrylhydrazyl (DPPH), ferric ion-chelating capacity and ferric-reducing antioxidant power (FRAP) assays at 250, 500, and 1000 μg mL⁻¹. Total phenol and flavonoid quantification of the extracts was achieved using Folin-Ciocalteau and AlCl₃ reagents, respectively. The ethyl acetate extract of C. arvensis flowers was the most active in AChE inhibition assay (31.24 ± 1.29%), while the n-hexane extract of C. officinalis leaves exerted the highest ferric ion-chelating capacity (74.27 ± 2.25%). Thin layer chromatographic analysis indicated presence of flavonoid and triterpene derivatives mainly in the extracts.     

Efficacy of medicinal plant extracts against Formosan subterranean termite, Coptotermes formosanus

The Formosan subterranean termite, Coptotermes formosanus Shiraki, is among the most devastating termite pests. Natural products derived from plant extracts were tested in a discovery programme for effective, environment friendly termite control agents. Screening for anti-termitic activity of plant extracts with some known medicinal attributes could lead to the discovery of new agents for termite control. The aim of this study was to investigate the anti-termitic activity of crude leaf hexane, ethyl acetate, acetone and methanol extracts of Andrographis lineata Wallich ex Nees. (Acanthaceae), Andrographis paniculata (Burm.f.) Wall. ex Nees. (Acanthaceae), Argemone mexicana L. (Papaveraceae), Aristolochia bracteolata Lam. (Aristolochiaceae), Datura metel L. (Solanaceae), Eclipta prostrata L. (Asteraceae), Sesbania grandiflora (L.) Pers. (Fibaceae) and Tagetes erecta L. (Compositae) against C. formosanus. An impregnated filter paper no-choice bioassay method was followed. All the crude extracts showed anti-termitic activity in a dose-dependent manner and exhibited a significant activity after 24 h and 48 h of exposure; the highest termite mortality was found in leaf hexane extract of A. bracteolata, ethyl acetate extract of A. paniculata, D. metel, E. prostrata, methanol extract of A. lineata and D. metel after 24 h (LD₅₀ = 363, 371, 298, 292, 358 and 317 ppm; LD₉₀ = 1433, 1659, 1308, 1538, 1703 and 1469 ppm), respectively. The hexane extract of T. erecta, acetone extract of A. mexicana, methanol extract of S. grandiflora and T. erecta showed activity after 48 h (LD₅₀ = 245, 253, 289, 409 ppm; LD₉₀ = 1378, 1511, 1508 and 2425 ppm), respectively. Among the natural products tested, may provide a renewable source of safe natural wood preservatives. These findings corroborate traditional insecticidal application of selected plants and the results can be extended for the control of termites. The primary objective of the present study was to identify novel, natural chemotypes from biologically active crude plant extracts that may be useful as part of termite treatment regimens in their natural form or as synthons for structure-activity studies in the future. The results reported here open the possibility of further investigations of efficacy on their anti-termitic properties of natural product extracts.     

Characterization of Solanum chomatophilum resistance to 2 aphid potato pests, Macrosiphum euphorbiae (Thomas) and Myzus persicae (Sulzer)

The aphids Macrosiphum euphorbiae (Thomas) and Myzus persicae (Sulzer) (Hemiptera: Aphididae) are responsible for yield reduction in potato (Solanum tuberosum) production by direct phloem feeding and by spreading viruses. Breeding resistant traits from Solanum chomatophilum into the potato germplasm provides alternative means to control aphid infestations. Integrated pest management strategy, using plant resistance, benefits from the characterization of the resistance and of its impact on aphid biology. Our objective was to characterize the resistance of S. chomatophilum by assessing the effects of accessions, plant parts on aphid performance, and by assessing the impact of the resistance factors on different aphid developmental stages and on alate morph production. Detailed aphid performance was obtained by measuring fecundity, survival, percentage of nymphs that reached adult moult, and population growth using whole plant and clip cage experimental designs. Accession and plant physiological age, but not aphid developmental stage, influenced all life-history parameters, except for alate morph production which was not induced on the resistant accessions. Plant part influence was independent of plant species and accession. Both experimental designs resulted in congruent resistance levels at the accession level for each of the two aphid species, supporting the use of any of them in S. chomatophilum resistance screening. PI243340 was resistant to both aphid species, while PI365324 and PI310990 were also resistant to M. euphorbiae and M. persicae, respectively.     

Potential of extracts of the tropical plant Balanites aegyptiaca (L) Del. (Balanitaceae) to control the mealy bug, Maconellicoccus hirsutus (Homoptera: Pseudococcidae)

The effects of extracts of different parts of the perennial tropical plant Balanites aegyptiaca (L) Del., including various solvent extracts of roots, methanol extracts from leaves, fruits, flowers and roots, partially purified saponins obtained from its roots and a standard saponin were studied on the life cycle (adult longevity, number of eggs, crawlers, adults, weight of adults and % wax content) of a laboratory-reared parthenogenic line of the mealy bug, Maconellicoccus hirsutus (Homoptera: Pseudococcidae). Extracts derived from various parts of B. aegyptiaca (leaves, fruits, flowers, and roots in methanol) affected the life cycle of M. hirsutus with a methanol root extract being the most effective at a concentration of 500 μg ml⁻¹. Partially purified saponin of B. aegyptiaca and the commercial bark saponin extract (Sigma) from Quillaja saponaria at a concentration of 500 μg ml⁻¹ were effective in reducing the longevity of M. hirsutus. Significant reductions in oviposition by M. hirsutus were found for all the extracts at a concentration of 500 μg ml⁻¹. Extracts also affected the number of emerging crawlers, number of adults as well as the weight and wax content of emerging adults. These studies suggest that B. aegyptiaca plant extracts and saponins can be useful botanical insecticides for the protection of crops from mealy bugs.     

Response of two tephritid species, Bactrocera oleae and Ceratitis capitata, to different emission levels of pheromone and parapheromone

Attractants and pheromones are commonly used in integrated pest management programs in crop systems. However, pheromone dispensers employed in monitoring traps and lure and kill devices are not usually well studied and attractants are released at uncontrolled rates leading to low treatment efficacies and misleading monitoring estimations. Fruit flies are pests of economic importance and monitoring is essential in order to program insecticidal treatments. Moreover, lure and kill techniques are being increasingly used, but the cost of these techniques depends on the number of required traps and, therefore, on the efficacy of the attractants. Ceratitis capitata and Bactrocera oleae are the two main fruit flies in Mediterranean countries, and the effect of different doses of trimedlure and spiroacetal on fly attraction has been studied. Results showed that a release rate over 1.28 mg/day of spiroacetal reduces B. oleae attraction and emission values over 2.4 mg of trimedlure per day did not increase C. capitata catches. Under the environmental conditions of our study, an optimum release rate for pheromone attraction in B. oleae was determined. Emission values over this optimum level reduced B. oleae attraction. However, when a parapheromone was used with C. capitata, a fruit fly of the same family, the optimum emission value was not found and higher quantities of parapheromone attracted the same number of flies. The saturation effect of high concentrations of pheromone and parapheromone is discussed.     

Comparative susceptibility of Ostrinia furnacalis, Ostrinia nubilalis and Diatraea saccharalis (Lepidoptera: Crambidae) to Bacillus thuringiensis Cry1 toxins

Transgenic corn hybrids that express toxins from Bacillus thuringiensis (Bt) are highly effective against the European corn borer, Ostrinia nubilalis (Hübner), and the closely related Asian corn borer, Ostrinia furnacalis (Guenée). Since the registration of Bt corn hybrids in the U.S. in 1996, there has been a great deal of information generated on O. nubilalis. However, relatively little information exists for O. furnacalis. To help determine whether the information generated for O. nubilalis can be leveraged for decisions regarding the use of transgenic Bt corn against O. furnacalis, experiments were designed to determine whether the pattern of sensitivity to various Bt Cry1 toxins is similar between the two species. Test insects included laboratory-reared O. furnacalis originating from Malaysia, a Bt-susceptible laboratory colony of O. nubilalis maintained at the University of Nebraska-Lincoln (UNL) and an out-group consisting of the sugarcane borer, Diatraea saccharalis (F.), from Louisiana which represents a different genus from the same family. O. furnacalis and O. nubilalis exhibited a similar pattern of susceptibility to all the Cry1 toxins and were highly susceptible to the range of Bt toxins tested including Cry1Aa, Cry1Ab, Cry1Ac and Cry1F. Both of the Ostrinia species were more tolerant to Cry1Ba compared with D. saccharalis, although sensitivity of O. furnacalis was intermediate and did not differ significantly from that of O. nubilalis and D. saccharalis. D. saccharalis was also susceptible to the range of toxins tested but unlike the two Ostrinia species, was more tolerant to Cry1F and more susceptible to Cry1Ba. These results indicate that both of the Ostrinia corn borer species are similar in sensitivity to the Cry1Aa, Cry1Ab, Cry1Ac, Cry1Ba and Cry1F toxins, thus suggesting shared toxin receptors and mechanisms of toxicity for the two species.     

Efficacy of plant extracts and biocontrol agents against Pythium aphanidermatum inciting chilli damping-off

The fungitoxic effects of 66 medicinal plants belonging to different families were evaluated in vitro on Pythium aphanidermatum, the causal agent of chilli damping-off. Of these, Zimmu leaf extract (Allium sativum L. × Allium cepa L.) showed the highest inhibition of mycelial growth of P. aphanidermatum (13.7 mm). The antimicrobial compounds were isolated from Zimmu leaf extract and 22 compounds were identified through gas chromatography mass spectroscopy (GC–MS). Biocontrol agents Trichoderma viride and Pseudomonas fluorescens and Zimmu extract were also tested alone and together in vitro and in vivo experiments for control of P. aphanidermatum. The in vitro studies revealed that combination of T. viride + P. fluorescens + Zimmu leaf extract showed the highest mycelial growth inhibition over the control. Both antagonists were compatible with each other and with Zimmu leaf extract. The pot culture studies revealed that seed treatment with combined application of T. viride + P. fluorescens + Zimmu leaf extract was superior in reducing the pre and post-emergence damping-off incidence (8.3 and 17.0%, respectively), and increased the plant growth and yield (shoot length and root length of 13.7 and 6.3 cm, 146 g/plant, respectively) of chilli when compared to control.     

Population dynamics of Xanthomonas campestris pv. vitians on different plant species and management of bacterial leaf spot of lettuce under greenhouse conditions

The population dynamics of Xanthomonas campestris pv. vitians (Xcv) was studied both externally and internally in lettuce, tomato and pepper plants. In addition, the application of bactericides during transplant production period was carried out for the management of bacterial leaf spot of lettuce under greenhouse conditions. Epiphytic populations of Xcv were recovered on leaves of lettuce plants (10⁵ CFU/g) 5 weeks after sprayed than the other plant species when inoculated with 10⁸ CFU/ml of Xcv. When plants of each crop species infiltrated with the bacterium at 10⁵ CFU/ml, the highest populations were developed in lettuce (10⁸ CFU/cm²) followed by pepper with 10⁶ CFU/cm² and tomato with 10⁵ CFU/cm² 10-days after infiltration. Application of a mixture of Maneb and Kocide or Kocide alone as a foliar spray on lettuce significantly reduced the incidence and disease severity of bacterial leaf spot by 29 and 27% respectively. Spread of the bacterium and development of the disease may partly be managed by avoiding intercropping of these plants commonly grown in close proximity to lettuce. For the management of leaf-associated populations of Xcv in lettuce, use of a mixture of Maneb and Kocide is advocated to minimize the effect of attacks.     

Quantification of Bacillus thuringiensis Cry1Ab protein in tissues of YieldGard (MON810) corn hybrids tested at multiple field locations in India

Transgenically expressed Bacillus thuringiensis insecticidal-protein Cry1Ab was quantified in target tissues of insect feeding of several YieldGard^® corn hybrids. The Cry1Ab protein is intended to protect corn plants from two economically important stem borers, Chilo partellus and Sesamia inferens. A total of seven YieldGard hybrids, all with MON810 event, were field-tested in a total of fourteen locations during the dry season (October–March) of 2005/2006 and wet season (May–October) of 2006. S. inferens and C. partellus oviposit on leaves of young corn plants, 15–60 days after emergence (DAE). The neonates initially feed by scraping the leaf lamina before migrating to bore into the stem. Thus high concentrations of Cry1Ab in whorl leaf and stem tissues would ensure effective control of the borers. The mean tissue Cry1Ab concentrations during the oviposition window of the borers (15–60 DAE), ranged from 50.05 to 21.01 ppm in whorl leaf, and between 9.26 and 3.47 ppm in stem tissue during the same period in the dry season of 2005/06. Similarly, Cry1Ab concentrations in whorl leaf and stem between 15 and 60 DAE during the wet season of 2006 ranged between 19.30 to 11.08 and 14.28 to 4.69 ppm, respectively. The baseline-sensitivity data of these insects to Cry1Ab in laboratory assays was determined. The concentrations of Cry1Ab in the target tissues as studied in seven YieldGard hybrids tested suggest effective management of the two borers. This paper also provides a summary of the expression of the Cry1Ab gene in various genetic backgrounds.     

Feeding perturbation and toxic activity of five Chrysanthemum species crude extracts against Spodoptera littoralis (Boisduval) (Lepidoptera; Noctuidae)

The effect of the whole methanol extracts of five Chrysanthemum species on feeding and performance of Spodoptera littoralis (Boisduval) larvae has been investigated in vitro. The extracts exhibited an anti-feeding and phagostimulating activities against cotton leafworm larvae when applied either on leaf discs or incorporated into an artificial diet. Under chosen conditions, the antifeedant index calculated over 24 h for sixth instar larvae significantly varied from 78.55 for Chrysanthemum segetum L. to −44.18 for Chrysanthemum fuscatum Desf. at the dose of 1000 ppm. Toxicity of the extracts was manifested by a high mortality, reduced growth rates, and low weight gain by larvae fed on diets containing 1000–10,000 ppm of the extract. Anyone of the larvae treated with Chrysanthemum macrotum (D.R.) Ball. leaves crude extract survived to pupation at the two higher concentrations. The time to pupation increased for Chrysanthemum grandiflorum flowers crude extract from 11.40 ± 0.93 to 28.93 ± 10.92 days as the extract concentration in the diet increased from 0 to 10,000 ppm. The ingestion of crude extract by the third instar larvae reduced significantly the consumption, growth and utilisation of the ingested and digested food, and reduced digestibility.     

Actinomycetes mediated biochemical responses in tomato (Solanum lycopersicum) enhances bioprotection against Rhizoctonia solani

Six actinomycetes isolates, namely Streptomyces toxytricini vh6, Streptomyces flavotricini vh8, S. toxytricini vh22, Streptomyces avidinii vh32, Streptomyces tricolor vh85 and vh41, an isolate of an unknown species of Actinomycetales, were tested for their efficacy in protecting tomato (Solanum lycopersicum) against Rhizoctonia solani under green house conditions. Actinomycetes treated plants showed better growth in terms of high chlorophyll content, higher phenylalanine ammonia lyase (PAL) activity and high total phenolic content. Qualitative and quantitative estimation of phenolic compounds from tomato leaves showed significant accumulation of six phenolic acids, gallic (29.02 μg g⁻¹ fresh leaf wt), ferulic (11.44 μg g⁻¹ fresh wt), cinnamic (56.84 μg g⁻¹ fresh wt), gentisic (24.19 μg g⁻¹ fresh wt), chlorogenic acid (1.72 μg g⁻¹ fresh wt) and salicylic (0.39 μg g⁻¹ fresh wt) acid, in actinomycetes treated plants. Biochemical profiling, when correlated with plant mortality in actinomycetes treated and untreated plants, indicated that isolates vh6 and vh8 offered 44.55% and 40.14% disease reductions, respectively compared to the control. These results established that these organisms have the potential to act as biocontrol agents.     

Management of the root-knot nematode Meloidogyne incognita on tomato with combinations of different biocontrol organisms

The nematicidal effect of Pseudomonas fluorescens, Paecilomyces lilacinus, Pichia guilliermondii and Calothrix parietina singly or in combination was tested against root-knot nematode, Meloidogyne incognita. Treatments with P. fluorescens and P. lilacinus caused mortality of M. incognita as 45% and 30% of juveniles after 48 h of exposures, respectively compared to water control in vitro. Under greenhouse conditions, all treatments reduced the disease severity and enhanced plant growth compared to untreated control. Application of P. fluorescens, P. lilacinus and P. guilliermondii Moh 10 was more effective compared to C. parietina. There was a negative interaction between C. parietina and either P. lilacinus or P. guilliermondii. Fresh and dry weight of shoots and roots of plants were significantly reduced as a result of infection with M. incognita, however application of biocontrol agents singly or in mix recovered this reduction. Moreover, they enhanced the growth parameters compared with the control. Our results proved that application of different biocontrol agents (P. fluorescens, P. lilacinus and P. guilliermondii) not only has a lethal effect on nematode, but also enhances the plant growth, supplying many nutritional elements and induction the systemic resistance in plants. Presence of C. parietina as a soil inhabitant cyanobacterium could antagonize biocontrol agents leading to the reduction of their practical efficiency in soil.     

Reduced height alleles (Rht) and Hagberg falling number of wheat

Near-isogenic lines varying for alleles for reduced height (Rht) and photoperiod insensitivity (Ppd-D1) in cv. Mercia (2005/6–2010/11; rht (tall), Rht-B1b, Rht-D1b, Rht-B1c, Rht8c+Ppd-D1a, Rht-D1c, Rht12) and cvs Maris Huntsman and Maris Widgeon (2007/8–2010/11; rht (tall), Rht-B1b, Rht-D1b, Rht-B1c, Rht-B1b+Rht-D1b, Rht-D1b+Rht-B1c) were compared at one field site, but within different systems (‘organic’, O, 2005/6–2007/8 v. ‘intensive’, I, 2005/6–2010/11). Further experiments at the site (2006/7–2008/9) compared 64 lines of a doubled-haploid (DH) population [Savannah (Rht-D1b) × Renesansa (Rht-8c+Ppd-D1a)]. Gibberellin (GA) insensitive dwarfing alleles (Rht-B1b; Rht-B1c; Rht-D1b; Rht-D1c) could reduce α-amylase activity and/or increase Hagberg falling number (HFN) but effects depended greatly on system, background and season. Only Rht-B1c increased grain dormancy despite producing plants taller than Rht-D1c. The GA-sensitive Rht8c+Ppd-D1a in Mercia was associated with reduced HFN but analysis of the DH population suggested this was more closely linked with Ppd-D1a, rather than Rht8c. The GA-sensitive severe-dwarfing allele Rht12 was associated with reduced HFN. Instability in HFN over season tended to increase with degree of dwarfing. There was a negative association between mean grain weight and HFN that was in addition to effects of Rht and Ppd-D1 allele.     

Insecticidal effect of recombinant endophytic bacterium containing Pinellia ternata agglutinin against white backed planthopper, Sogatella furcifera

White backed planthopper (WBPH; Sogota furcifera Horvath) has become the major threat to rice crops throughout Asia, damaging plants both through its feeding behavior and by acting as a virus vector. Here, we developed a novel method for biologically controlling WBPH by using endophytic bacterium to express anti-pest plant lectins. Strain SJ-10 of an endophytic bacterium, characterized as Enterobacter cloacae by morphological, physiological, biochemical and 16s rDNA characteristics, was isolated from rice seedlings. The Pinellia ternate agglutinin (PTA) gene was cloned into SJ-10 for expression. The positive transformant, selected by antibiotic resistance, was evaluated using PCR, SDS-PAGE and Western blot assay. After inoculation, rSJ-10 could colonize rice plants so that they expressed PTA, and then the rice was shown to have insecticidal activity against WBPH. The results showed that rSJ-10 could significantly decrease the survival and fecundity of WBPH fed on rice seedlings (p < 0.01). At day 19, the fecundity of WBPH inoculated with rSJ-10, or with wild-type SJ-10 was decreased by 86.1%, and 25.6%, respectively. At day 22, numbers of WBPH on rice in the control were 19.4 times greater than on rice inoculated with rSJ-10. At day 26, the rice seedlings all died in the control group, but the seedlings inoculated with rSJ-10 grew well. The results showed that the rice seedlings inoculated with rSJ-10 expressing PTA protein were endowed with the anti-pest activity against WBPH. Further work is needed to investigate whether the rice plants expressing rPTA are toxic to mammals. This research highlights a way to biologically control planthoppers by recombinant endophytic bacteria expressing plant lectins.