Metabolism of [ring-2,6-14]parathion in plant cell suspension cultures of carrot (Daucus carota), purple foxglove (Digitalis purpurea), soybean,thorn apple (Datura stramonium) and wheat (Triticum aestivum)

By means of standardized procedures, the metabolism of [ring-2,6-¹⁴C]-parathion was investigated in carrot (Daucus carota L.), purple foxglove (Digitalis purpurea L.), soybean (Glycine max Merrill cv. ?Mandarin’?, and Glycine max Merrill cv. ?Harosoy 63’? cultivated on B5 and Miller media, respectively), thorn apple (Datura stramonium L.), and wheat (Triticum aestivum L.) cell suspension cultures. In the wheat and soybean (Mandarin) cells only 2.9 and 8.9%, respectively, of the applied parthion remained unmetabolized after 48 h of incubation, while 51.2, 57.9, 60.3, and 62.4% of the unchanged parent were detected in the D. purpurea, D. Stramonium, carrot and soybean (Harosoy) cultures, respectively. In all suspensions, paraoxon and 4-nitrophenol were found as phase I metabolites, thus demonstrating that plant tissues can catalyse oxidative desulfuration and dearylation of parathion. 4-Nitrophenol was also glycosylated with glucose and possibly galactose. Further, as yet unidentified, metabolites indicated that bio-transformations had also occurred at the aromatic moiety. Large amounts of non-extractable residues were detected in the wheat suspension (38.3%), while the other cultures showed a lower incorporation of ¹⁴C into insoluble cell material (0.9-9.4%). For a prospective ecotoxicological evaluation of the metabolic fate of pesticides and xenobiotics in plants in general, the differential metabolic capacity of plant cell cultures and plants should be taken into account.     

Increased activity and reduced sensitivity of glutamine synthetase in glufosinate‐resistant vetiver (Vetiveria zizanioides Nash) cells

Vetiver (Vetiveria zizanioides Nash) cells derived from an inflorescence were cultured in a modified N6 liquid medium supplemented with 10 µm 2,4‐D and 10 mm proline. Exponentially growing cell suspensions were subcultured with a selection medium containing glufosinate (ammonium dl ‐homoalanin‐4‐yl(methyl)phosphinate). The glufosinate‐resistant cells which can grow in a medium containing 5 × 10^?5 M glufosinate was selected by a stepwise selection, and its I₅₀ value was determined to be 4.2 × 10^?5 M. The growth of susceptible cells was inhibited by lower concentrations of glufosinate and its I₅₀ value was 2.5 × 10^?7 M. This indicated that the selected cells were 170‐fold resistant compared with the susceptible cells. Glutamine synthetase (GS) activity of the resistant cells was twice as high as that of the susceptible cells. The I₅₀ values of glufosinate were 3.2 × 10^?5 M and 9.0 × 10^?7 M for GS from the resistant and susceptible cells, respectively. The accumulation of ammonia caused by GS inhibition was higher in the susceptible cells. Absorption of [3,4–¹⁴C]glufosinate was not significantly different between the resistant and susceptible cells. Both cell types did not metabolize glufosinate. These results suggest that the resistance of the selected vetiver cell suspension to glufosinate is mainly due to increased GS activity and its decreased sensitivity to the herbicide.     

Insecticidal action of mammalian galectin‐1 against diamondback moth (Plutella xylostella)

BACKGROUND: Previous studies showed that mammalian galectin‐1 (GAL1) could interact with chitosan or chitin, one component of the peritrophic membrane (PM). This finding suggests that the PM could be a target of GAL1, which prompted the authors to explore the effect of GAL1 on larval growth and its potential mechanism. RESULTS: The development of Plutella xylostella (L.) larvae was significantly disturbed after they were fed recombinant GAL1. The histochemical structure and immunostaining pattern suggested that GAL1 treatment resulted in dose‐ and time‐dependent disruption of the microvilli and abnormalities in these epithelial cells. Ultrastructural studies showed that the PM was not present in the midgut of GAL1‐treated insects; instead, numerous bacteria were found in the lumen area. These results indicate that the protective function of the PM was disrupted by GAL1 treatment. Moreover, in vitro data showed that GAL1 interacts with chitosan/chitin in a dose‐dependent manner, and also specifically binds to the PM in vitro. CONCLUSION: In view of the fact that the carbohydrate recognition domain of GAL1 recognises the structural motif N‐acetyl lactosamine (Gal β 1–4 GlcNAc), which is similar to that of chitin (β‐1,4 N‐acetyl‐D ‐glucosamine), it is proposed that the insecticidal mechanism of GAL1 involves direct binding with chitin to interfere with the structure of the PM. Copyright © 2009 Society of Chemical Industry     

Potential gene flow from transgenic rice (Oryza sativa L.) to different weedy rice (Oryza sativa f. spontanea) accessions based on reproductive compatibility

BACKGROUND: The possibility of gene flow from transgenic crops to wild relatives may be affected by reproductive capacity between them. The potential gene flow from two transgenic rice lines containing the bar gene to five accessions of weedy rice (WR1–WR5) was determined through examination of reproductive compatibility under controlled pollination. RESULTS: The pollen grain germination of two transgenic rice lines on the stigma of all weedy rice, rice pollen tube growth down the style and entry into the weedy rice ovary were similar to self‐pollination in weedy rice. However, delayed double fertilisation and embryo abortion in crosses between WR2 and Y0003 were observed. Seed sets between transgenic rice lines and weedy rice varied from 8 to 76%. Although repeated pollination increased seed set significantly, the rank of the seed set between the weedy rice accessions and rice lines was not changed. The germination rates of F₁ hybrids were similar or greater compared with respective females. All F₁ plants expressed glufosinate resistance in the presence of glufosinate selection pressure. CONCLUSIONS: The frequency of gene flow between different weedy rice accessions and transgenic herbicide‐resistant rice may differ owing to different reproductive compatibility. This result suggests that, when wild relatives are selected as experimental materials for assessing the gene flow of transgenic rice, it is necessary to address the compatibility between transgenic rice and wild relatives. Copyright © 2009 Society of Chemical Industry     

Development of insect‐resistant transgenic rice with Cry1C*‐free endosperm

BACKGROUND: Yellow stem borer (Tryporyza incertulas Walker), striped stem borer (Chilo suppressalis Walker) and leaf folder (Cnaphalocrocis medinalis Guenec) are three lepidopteran pests that cause severe damage to rice in many areas of the world. In this study, novel insect‐resistant transgenic rice was developed in which Bt protein expression was nearly absent in the endosperm. The resistant gene, cry1C*, driven by the rice rbcS promoter (small subunit of ribulose‐1,5‐bisphosphate carboxylase/oxygenase), was introduced into Zhonghua 11 (Oryza sativa L. ssp. japonica) by Agrobacterium‐mediated transformation. RESULTS: A total of 83 independent transformants were obtained, 19 of which were characterised as single‐copy foreign gene insertion. After preliminary screening of the T₁ families of these 19 transformants in the field, six highly insect‐resistant homozygous lines were selected. These six homozygous transgenic lines were field tested for resistance to leaf folders and stem borers, and for their agronomic performance. The Cry1C* protein levels in leaves and endosperm were measured by ELISA. Subsequently, the elite transgenic line RJ5 was selected; this line not only possessed high resistance to leaf folders and stem borers, normal agronomic performance, but also Cry1C* expression was only 2.6 ng g^?1 in the endosperm. CONCLUSION: These results indicated that RJ5 has the potential for widespread utility in rice production. Copyright © 2009 Society of Chemical Industry     

Preliminary findings of potentially resistant goosegrass (Eleusine indica) to glufosinate‐ammonium in Malaysia

Goosegrass (Eleusine indica), regarded as one of the world's worst weeds, is highly pernicious to cash crop‐growers in Malaysia. Following reports in 2009 that glufosinate‐ammonium failed to adequately control goosegrass populations in Kesang, Malacca, and Jerantut, Pahang, Malaysia, on‐site field trials were conducted to assess the efficacy of glufosinate‐ammonium towards goosegrass in both places. Preliminary screenings with glufosinate‐ammonium at the recommended rate of 495 g ai ha^?1 provided 82% control of the weed at a vegetable farm in Kesang, while the same rate failed to control goosegrass at an oil palm nursery in Jerantut. The ensuing greenhouse evaluations indicated that the “Kesang” biotype exhibited twofold resistance, while the “Jerantut” biotype exhibited eightfold resistance towards glufosinate‐ammonium, compared to susceptible goosegrass populations. The occurrence of glufosinate resistance in goosegrass calls for more integrated management of the weed to prevent escalating resistance and further proliferation of the weed in Malaysia.     

Structure–biological activity relationships in triterpenic saponins: the relative activity of protobassic acid and its derivatives against plant pathogenic fungi

BACKGROUND: Triterpenic saponins from Sapindus mukorossi Gaertn. and Diploknema butyracea JF Gmelin were evaluated for in vitro antifungal activity against four phytopathogenic fungi. The study of the structure–antifungal activity relationships of protobassic acid saponins was widened by including semi‐synthetic derivatives. RESULTS: Diploknema butyracea saponins exhibited significant antifungal activity against three fungi (ED₅₀ 230–455 µg mL^?1), whereas S. mukorossi saponin was effective against two fungi (ED₅₀ 181–407 µg mL^?1). The n‐butanol extract after preparative HPLC separation provided two saponins from D. butyracea saponin mixture: 3‐O‐[β‐D ‐glucopyarnosyl‐β‐D ‐glucopyranosyl]‐16‐α‐hydroxyprotobassic acid‐28‐O‐[arabinopyranosyl‐glucopyranosyl‐xylopyranosyl]‐arabinopyranoside (MI‐I), and 3‐O‐β‐D ‐glucopyranosyl‐glucopyranosyl‐glucopyranosyl‐16‐α‐hydroxyprotobassic acid‐28‐O‐[arabinopyranosyl‐xylopyranosyl‐arabinopyranosyl]‐apiofuranoside (MI‐III). The single saponin extracted from S. mukorossi saponin mixture was identified as 3‐O‐[O‐acetyl‐β‐D ‐xylopyranosyl‐β‐D ‐arabinopyranosyl‐β‐D ‐rhamnopyranosyl] hederagenin‐28‐O[β‐D ‐glucopyranosyl‐β‐D ‐glucopyranosyl‐β‐D ‐rhamnopyranosyl] ester (SM‐I). Monodesmosides resulting from the partial degradation of hederagenin and hydroxyprotobassic acid bisdesmosides exhibited significant reduction in antifungal effect. Further removal of sugar moiety yielded complete loss in activity. The antifungal activity of the triterpenic saponins was associated with their aglycone moieties, and esterification of the hydroxyl group led to change in antifungal activity. CONCLUSION: Sapindus mukorossi saponin, which is effective against Rhizoctonia bataticola (Taub.) Briton Jones and Sclerotium rolfsii Sacc., can be exploited for the development of a natural fungicide. A sugar moiety is a prerequisite for the antifungal activity of triterpenic saponin. Copyright © 2010 Society of Chemical Industry     

草铵膦对转基因水稻品系99-1和普通水稻品种的无影响浓度比较

在室内采用水培方法,比较草铵膦对抗草铵膦转基因水稻品系99-1与普通水稻品种越富和中作93的无影响浓度,从而明确该除草剂对水稻是否存在残留药害.结果表明,草铵膦对转基因水稻品系99-1的无影响浓度明显高于非转基因品种,对前者的种子和2叶龄幼苗的无影响浓度分别为8.28 mg/L和48.97 mg/L,而对后者的种子和2叶龄幼苗的无影响浓度分别为1.83 mg/L和2.06 mg/L.这些无影响浓度均高于推荐用量下草铵膦在土壤中的残留浓度.因此,正常用量下的草铵膦对转基因水稻品系和普通水稻均无土壤残留问题.     

Perspectives on transgenic,herbicide‐resistant crops in the United States almost 20 years after introduction

Herbicide‐resistant crops have had a profound impact on weed management. Most of the impact has been by glyphosate‐resistant maize, cotton, soybean and canola. Significant economic savings, yield increases and more efficacious and simplified weed management have resulted in widespread adoption of the technology. Initially, glyphosate‐resistant crops enabled significantly reduced tillage and reduced the environmental impact of weed management. Continuous use of glyphosate with glyphosate‐resistant crops over broad areas facilitated the evolution of glyphosate‐resistant weeds, which have resulted in increases in the use of tillage and other herbicides with glyphosate, reducing some of the initial environmental benefits of glyphosate‐resistant crops. Transgenic crops with resistance to auxinic herbicides, as well as to herbicides that inhibit acetolactate synthase, acetyl‐CoA carboxylase and hydroxyphenylpyruvate dioxygenase, stacked with glyphosate and/or glufosinate resistance, will become available in the next few years. These technologies will provide additional weed management options for farmers, but will not have all of the positive effects (reduced cost, simplified weed management, lowered environmental impact and reduced tillage) that glyphosate‐resistant crops had initially. In the more distant future, other herbicide‐resistant crops (including non‐transgenic ones), herbicides with new modes of action and technologies that are currently in their infancy (e.g. bioherbicides, sprayable herbicidal RNAi and/or robotic weeding) may affect the role of transgenic, herbicide‐resistant crops in weed management. Published 2014. This article is a U.S. Government work and is in the public domain in the USA.     

Natural tolerance of Cuscuta campestris to herbicides inhibiting amino acid biosynthesis

The response of Cuscuta campestris Yuncker, a non‐specific above‐ground holoparasite, to amino acid biosynthesis inhibitor (AABI) herbicides, was compared with other resistant and sensitive plants in dose–response assays carried out in Petri dishes. Cuscuta campestris was found to be much more resistant to all AABI herbicides tested. The I₅₀ value of C. campestris growth inhibition by glyphosate was eightfold higher than that of transgenic, glyphosate‐resistant cotton (RR‐cotton). The I₅₀ value for C. campestris shoot growth inhibition by sulfometuron was above 500 μM, whereas that of sorghum roots was only 0.004 μM. Cuscuta campestris exposed to glyphosate gradually accumulated shikimate, confirming herbicide penetration into the parasite and interaction with an active form of the target enzyme of the herbicide, 5‐enolpyruvylshikimate‐3‐phosphate synthase. More than half of the C. campestris plants associated with transgenic, glyphosate‐resistant sugarbeet (RR‐sugarbeet) treated with glyphosate or with transgenic, sulfometuron‐resistant tomato (SuR‐tomato) treated with sulfometuron recovered and resumed regular growth 20–30 days after treatment. New healthy stems developed, followed by normal flowering and seed setting. The results of the current study demonstrate the unique capacity of C. campestris to tolerate high rates of AABI. The mechanism of this phenomenon is yet to be elucidated.     

The glutamate dehydrogenase gene gdhA increased the resistance of tobacco to glufosinate

The gene gdhA from Escherichia coli, that encodes a NADPH‐dependent glutamate dehydrogenase (GDH), directs a novel pathway in transgenic plants that allows an increase in ammonium assimilation. Glufosinate leads to plant death by the irreversible inhibition of glutamate synthetase (GS) leading to a disruption of subsequent GS‐related processes resulting in elevated ammonium and disruption of photorespiration. Therefore, it was speculated that the gdhA‐transformed plants may exhibit a novel mechanism of resistance to glufosinate by altered activity of the GDH‐directed pathway(s) and subsequently related processes. Studies were conducted in the greenhouse to evaluate the resistance of tobacco plants containing the gdhA gene to glufosinate. Five tobacco genotype lines were investigated including a non‐transformed control line, a positive control line and three transformed lines with levels of increasing GDH activity directed by the gdhA gene. Plants transformed with the gdhA gene expressed up to six times increased level of resistance (GR₅₀) to glufosinate compared with the non‐transformed control, which is 100 times less resistant than plants transformed with the bar gene. The GDH activity among lines was highly correlated (r² = 0.9903) with the level of herbicide resistance. Thus, the use of the E. coli gdhA gene in plant transformations can provide an additional mechanism for resistance to glufosinate.     

Adverse effect of the chitinolytic enzyme PjCHI‐1 in transgenic tomato on egg mass production and embryonic development of Meloidogyne incognita

A novel chitinase gene (PjCHI‐1) isolated from Paecilomyces javanicus, a non‐nematophagous fungus, and driven by a CaMV35S promoter, was delivered into CLN2468D, a heat‐tolerant cultivar of tomato (Solanum lycopersicum). T₁ tomato plants exhibited high endochitinase activity and reduced numbers of eggs and egg masses when infected with the root‐knot nematode (RKN) Meloidogyne incognita. The eggs found in transgenic tomato had lower shell chitin contents than eggs collected from control plants. Egg masses from transgenic plants exhibited higher chitinase activity than those from control plants. Moreover, only 30% of eggs from transgenic plants were able to develop to the multi‐cell/J1 stage, compared with more than 96% from control plants. The present study demonstrated that the expression of the PjCHI‐1 chitinase gene can effectively reduce the production of egg masses and repress the embryonic development of M. incognita, presenting the possibility of a novel agro‐biotechnological strategy for preventing crop damage by RKN.     

Multiple resistance across glufosinate,glyphosate, paraquat and ACCase‐inhibiting herbicides in an Eleusine indica population

An Eleusine indica population was previously reported as the first global case of field‐evolved glufosinate resistance. This study re‐examines glufosinate resistance and investigates multiple resistance to other herbicides in the population. Dose–response experiments with glufosinate showed that the resistant population is 5‐fold and 14‐fold resistant relative to the susceptible population, based on GR₅₀ and LD₅₀ R/S ratio respectively. The selected glufosinate‐resistant subpopulation also displayed a high‐level resistance to glyphosate, with the respective GR₅₀ and LD₅₀ R/S ratios being 12‐ and 144‐fold. In addition, the subpopulation also displayed a level of resistance to paraquat and ACCase‐inhibiting herbicides fluazifop‐P‐butyl, haloxyfop‐P‐methyl and butroxydim. ACCase gene sequencing revealed that the Trp‐2027‐Cys mutation is likely responsible for resistance to the ACCase inhibitors examined. Here, we confirm glufosinate resistance and importantly, we find very high‐level glyphosate resistance, as well as resistance to paraquat and ACCase‐inhibiting herbicides. This is the first confirmed report of a weed species that evolved multiple resistance across all the three non‐selective global herbicides, glufosinate, glyphosate and paraquat.     

Acaricidal activity of triketone analogues derived from Leptospermum scoparium oil against house‐dust and stored‐food mites

BACKGROUND: Various attempts to control the populations of house‐dust and stored‐food mites have been implemented using synthetic chemicals. Although effective, the repeated use of these chemicals has led to resistance owing to the mite's high reproductive potential and short life cycle. Therefore, this study aimed to develop natural acaricides using oils derived from Leptospermum scoparium JR & G Forst., which may affect the overall biological activity of a mite without adverse effects. Results were compared with those from using benzyl benzoate and N,N‐diethyl‐3‐methylbenzamide (DEET). RESULTS: The LD₅₀ values of L. scoparium oil were 0.54, 0.67 and 1.12 µg cm^?2 against Dermatophagoides farinae (Hughes), D. pteronyssinus (Troussart) and Tyrophagus putrescentiae (Schrank) respectively. The active constituent isolated from L. scoparium was identified as leptospermone (6‐isovaleryl‐2,2,4,4‐tetramethyl‐1,3,5‐cyclohexanetrione) by spectroscopic analysis. Based on the LD₅₀ values of leptospermone and its derivatives, the most toxic compound against D. farinae was leptospermone (0.07 µg cm^?2), followed by 2,2,4,4,6,6‐hexamethyl‐1,3,5‐cyclohexanetrione (1.21 µg cm^?2), benzyl benzoate (10.03 µg cm^?2) and DEET (37.12 µg cm^?2). Furthermore, similar results were observed when the leptospermone and its derivatives were tested against D. pteronyssinus and T. putrescentiae. CONCLUSIONS: These results indicate that L. scoparium oil‐derived materials, particularly leptospermone and 2,2,4,4,6,6‐hexamethyl‐1,3,5‐cyclohexanetrione, have potential for development as new agents for the control of three species of mite. Copyright © 2008 Society of Chemical Industry     

Acaricidal activities of the active component of Lycopus lucidus oil and its derivatives against house dust and stored food mites (Arachnida: Acari)

BACKGROUND: Recent studies have focused on materials derived from plant extracts as mite control products against house dust and stored food mites because repeated use of synthetic acaricides had led to resistance and unwanted activities on non‐target organisms. The aim of this study was to evaluate the acaricidal activity of materials derived from Lycopus lucidus against Dermatophagoides farinae, D. pteronyssinus and Tyrophagus putrescentiae. RESULTS: The LD₅₀ values of L. lucidus oil were 2.19, 2.25 and 8.45 µg cm^?2 against D. farinae, D. pteronyssinus and T. putrescentiae. The acaricidal constituent of L. lucidus was isolated by chromatographic techniques and identified as 1‐octen‐3‐ol. In a fumigant method against D. farinae, the acaricidal activity of 1‐octen‐3‐ol (0.25 µg cm^?2) was more toxic than N,N‐diethyl‐m‐toluamide (DEET) (36.84 µg cm^?2), followed by 3,7‐dimethyl‐1‐octen‐3‐ol (0.29 µg cm^?2), 1‐octen‐3‐yl butyrate (2.32 µg cm^?2), 1‐octen‐3‐yl acetate (2.42 µg cm^?2), 3,7‐dimethyl‐1‐octene (9.34 µg cm^?2) and benzyl benzoate (10.02 µg cm^?2). In a filter paper bioassay against D. farinae, 1‐octen‐3‐ol (0.63 µg cm^?2) was more effective than DEET (20.64 µg cm^?2), followed by 3,7‐dimethyl‐1‐octen‐3‐ol (1.09 µg cm^?2). CONCLUSION: 1‐Octen‐3‐ol and 3,7‐dimethyl‐1‐octen‐3‐ol could be useful as natural agents for the management of three mite species. Copyright © 2011 Society of Chemical Industry     

Increased resistance to fusarium wilt in transgenic tobacco lines co‐expressing chitinase and wasabi defensin genes

Marker‐free transgenic tobacco (Nicotiana tabacum) lines containing a chitinase (ChiC) gene isolated from Streptomyces griseus strain HUT 6037 were produced by Agrobacterium‐mediated transformation. One marker‐free transgenic line, TC‐1, was retransformed with the wasabi defensin (WD) gene, isolated from Wasabia japonica. Of the retransformed shoots, 37% co‐expressed the ChiC/WD genes, as confirmed by western and northern analyses. Southern blot analysis showed that no chromosomal rearrangement was introduced between the first and the second transformation. Transgenic lines either expressing ChiC or WD, or co‐expressing both genes were challenged with Fusarium oxysporum f.sp. nicotianae (Fon). Assessment of in vitro plant survival in the presence of Fon showed that transgenic lines co‐expressing both genes had significantly enhanced protection against the fungus (infection indices 0·0–1.·2) compared with corresponding isogenic lines expressing either of the genes (infection indices 2·5–9·8). Whole‐plant infection indices in transgenic lines were significantly related (r = 0·93, P < 0·01) to the extent of root colonization of the host, which ranged from 2·1% to 11·3% in lines co‐expressing both genes, and from 16·8% to 37·7% in lines expressing just one of the genes (compared with 86·4% in non‐transformed controls). Leaf extracts of transgenic lines also inhibited mycelial growth of Fon in vitro and caused hyphal abnormalities.     

Generation and evaluation of movement protein‐specific single‐chain antibodies for delaying symptoms of Tomato spotted wilt virus infection in tobacco

This study investigated whether single‐chain antibodies (scFvs) specific for a viral movement protein could accumulate in the plant cell cytosol and restrict viral systemic infection in plants. Nine chicken scFv fragments against the Tomato spotted wilt virus (TSWV) movement protein (NS_M) were isolated by phage display. Soluble scFvs were produced in bacteria and the NS_M binding activity of purified scFvs was confirmed. The nine scFv genes were cloned into a plant expression vector enabling recombinant protein accumulation in the plant cell cytosol. Immunoblot analysis demonstrated that two of the nine chicken scFvs accumulated to high levels (5·9 and 8·0% of total soluble protein). Bioassays of viral infection using transgenic tobacco plants producing NS_M‐specific chicken scFvs showed delayed symptom development when compared to non‐transgenic control plants, indicating that expression of antibodies recognizing the TSWV movement protein is a potential strategy for generating resistant plants.     

Silencing of Erwinia amylovora sy69 AHL‐quorum sensing by a Bacillus simplex AHL‐inducible aiiA gene encoding a zinc‐dependent N‐acyl‐homoserine lactonase

Quorum sensing in Gram‐negative bacteria is regulated by diffusible signal molecules called N‐acyl‐l ‐homoserine lactones (AHLs). These molecules are degraded by lactonases. In this study, six Bacillus simplex isolates were characterized and identified as a new quorum‐quenching species of Bacillus. An aiiA gene encoding an AHL‐lactonase was identified based on evidence that: (i) it showed high homology with other aiiA genes of Bacillus sp.; (ii) the deduced amino acid sequence contained two conserved regions, ¹⁰⁴SHLHFDH¹¹¹ and ¹⁶⁵TPGHTPGH¹⁷³, characteristic of the metallo‐β‐lactamase superfamily; and (iii) the protein had zinc‐dependent AHL‐degrading activity. Additionally, the expression of the aiiA gene was significantly up‐regulated by 3‐oxo‐AHL. The AHL‐lactonase inhibited multiplication of the 3‐oxo‐C6‐AHL‐producing plant pathogen Erwinia amylovora sy69 both in vitro and in planta. The results provide support for the use of the quorum‐quenching functionality of B. simplex in the integrated control of the devastating fire blight pathogen.     

Effect of dietary cowpea trypsin inhibitor (CpTI) on the growth and development of the tomato moth Lacanobia oleracea (Lepidoptera: Noctuidae) and on the success of the gregarious ectoparasitoid Eulophus pennicornis (Hymenoptera: Eulophidae)

Cowpea trypsin inhibitor (CpTI) was shown to have a deleterious effect on the growth and development of larvae of the tomato moth, Lacanobia oleracea, when incorporated in artificial diet (2.0 % of soluble protein) and expressed in transgenic potato leaf (up to 1.0 % of soluble protein). The effect of CpTI on parasitism of L oleracea by the ectoparasitoid Eulophus pennicornis was investigated. The parasitic success of the wasp was reduced by the presence of CpTI in the diet of the host and, in the case of transgenic potato leaves expressing the transgene protein, was collated with the length of time the host fed on the diet prior to parasitism. In all cases the proportion of hosts parasitised when fed CpTI‐containing diets was reduced when compared with controls, although these differences were only significant when hosts were fed from the third instar on the transgenic potato leaves. Parasitoid progeny that developed on L oleracea reared on CpTI‐containing diets, however, were not adversely affected. These results show that, whilst expression of CpTI in transgenic potato plants confers resistance to the lepidopterous pest L oleracea, adverse effects on the ability of the ectoparasitoid E pennicornis to parasitise this moth species successfully may also occur. These results are discussed in relation to the potential impact of transgenic crops on beneficial biological control agents. © Crown copyright 2001. Reproduced with the permission of Her Majesty's Stationery Office. Published for SCI by John Wiley & Sons, Ltd.     

Resistance to Rhizoctonia solani AG‐2‐2 (IIIB) in creeping bentgrass plants transformed with pepper esterase gene PepEST

A pepper esterase (PepEST) gene was introduced into creeping bentgrass (Agrostis stolonifera) by Agrobacterium‐mediated transformation. Purified recombinant PepEST proteins were sufficient to inhibit the growth of the fungal pathogens Rhizoctonia solani AG2‐2 (IIIB) (causing brown patch) and Sclerotinia homoeocarpa (dollar spot), but not the oomycete responsible for pythium blight, Pythium aphanidermatum. PepEST proteins were most effective against R. solani. After genetic transformation of creeping bentgrass with PepEST, the genomic integration of transgenes bar and PepEST was confirmed by Southern blot analysis, and their expression was also validated by northern blot and western blot analyses. Disease severity on R. solani‐inoculated leaves of transgenic plants was <10% compared to ca. 50% in non‐transgenic plants. Microscopic observation of infected leaves indicated that PepEST inhibited the growth of hyphae upon fungal infection.