Potentialities of methoxyfenozide for the integrated management of Helicoverpa armigera (Lepidoptera: Noctuidae) on cotton in Benin,West Africa

Intensive use of chemical insecticides against Helicoverpa armigera has led to the development of resistance to the major chemical families of insecticides. Consequently, management of H. armigera using conventional chemical insecticides is increasingly difficult. Methoxyfenozide is an agonist of the insect moulting hormone 20-hydroxyecdysone that acts faster than chitin synthesis inhibitors. The present work is aimed to assess the effectiveness of methoxyfenozide for use against H. armigera on cotton in Benin, West Africa. Laboratory tests and field experiments have been carried out. For laboratory studies, topical application of methoxyfenozide was done using L2 and L5 instar larva of H. armigera. Tested methoxyfenozide concentrations varied from 24 μl/ml to 144 μl/ml and the control is water only. For field experiments, a complete randomised block design was used with methoxyfenozide (72 μl/ml) and a control (no spraying). Berthoud Ultra Low Volume sprayer was used to spray methoxyfenozide suspension at 60-l per hectare. Two sprays were applied, 7 days apart. Laboratory tests indicated that 24 h after application of methoxyfenozide, about 100% of treated L2 and L5 larva were morbid or dead compared with 0% for the control larva. There are no significant differences between tested concentrations. Morbid larvae died within 2 days. In field experiments, cotton yield harvested on methoxyfenozide treated plants was double of that obtained on untreated plants (9375 kg and 4875 kg per hectare respectively). Thus methoxyfenozide may be used as component of Integrated Pest Control Programme on cotton in Benin, West Africa.     

Clonostachys rosea,a potential biological control agent for Rhizoctonia solani AG-3 causing black scurf on potato

Rhizoctonia solani AG-3 forms sclerotia on potato tubers, reducing quality and marketability. Potato discs with sclerotia were exposed to saturated soil, and following toothpick baiting, hyphae of R. solani parasitised with mycelia that either coiled around or penetrated cell walls were observed. Similar parasitic behaviour was observed in dual cultures. Clonostachys rosea was identified as the mycoparasite by morphology and ITS sequencing. When co-inoculated onto stems grown from disease-free seed potatoes, tubers of the C. rosea?+?R. solani inoculated plants had significantly lower black scurf (P?0.0001) and higher yield (P?=?0.0002) than R. solani inoculated controls.     

Virulence potential of some fungal isolates and their control-promise against the Egyptian cotton leaf worm,Spodoptera littoralis

A preliminary virulence test of four fungal isolates, Beauveria bassiana IMI 382302, Beauveria bassiana IMI 386701, Trichoderma harzianum T24 and Aspergillus flavus Link against larvae of Spodoptera littoralis was performed. The most effective isolates against larvae of S. littoralis were B. bassiana 302 and T. harzianum T24, which also showed the lower percentage of pupation compared with the other two isolates under the same conditions of treatments. Three concentrations (1 × 10⁶, 1 × 10⁷ and 1 × 10⁸ ml^?1) of the aqueous conidial suspension of the four tested isolates were carried out against both larval and pupal stages of S. littoralis within five days post-treatment. T. harzianum T24 showed 80% larval mortality only when applied at the highest conidial concentration, while A. flavus showed 100% pupal mortality only, at all of its conidial concentrations. However, B. bassiana IMI 382302 showed relatively high dose-dependant larval and pupal mortalities, while strain IMI 386701 of B. bassiana showed a very weak mortality against pupae at higher concentrations, and no virulence against larvae was recorded. Enzymatic and antibiosis bioassays of the four fungal isolates showed relatively high activities against Fusarium spp. for most of the tested isolates. Clear zone of enzyme activity on agar plates proportionally increased with increasing the concentration of enzyme substrate and prolongation of the incubation period. Mtabolites produced in the agar culture inhibited the growth of Fusarium spp. and the productivity differed greatly among isolates or strains of the same isolate. Volatile and non-volatile compounds produced by A. flavus Link showed a higher inhibition activity against Fusarium spp. compared with the other fungal isolates. The humoral antifungal response of insect host is relatively high compared to the anti-bacterial one. Injection of larvae with the immune sensitive bacteria Micrococcus luteus (5 × 10³ bacteria/larva) showed a detectable humoral response by 2 h, peaked around 12 h and became hardly detectable by 24 h post-injection. Injection of larvae with conidial suspension (5 × 10³ conidia/larva) from each of the fungal isolates showed humoral antifungal activity against B. bassiana IMI 386701 and A. flavus only. This activity was detectable by 12 h, peaked around 36 h and became hardly detectable by 48 h post-injection. Although the humoral antifungal response was started slowly compared to the antibacterial one, it lasted for longer and enabled larvae to withstand the infection with these immune-sensitive fungal strains. No humoral activity was detected against B. bassiana IMI 382302, although however, weak activity was detected against T. harzianum T24 only at the low conidial concentration but not at the higher one (1 × 10⁸ ml^?1). Thus, this study concludes that larvae of S. littoralis showed immune-dependant sensitivity to T. harzianum T24 and B. bassiana IMI 382302. Therefore, this study may recommend these two fungal isolates as mycoinsecticides in the battle against cotton leaf worm in Egypt. Hence, they have been selected for future comprehensive bioassays in the laboratory under conditions similar to that in the field. This, in fact, may help for developing effective mycoinsecticides against this pest. Penetration mechanims of insect cuticle by entomopathogenic fungi will be discussed.     

Effect of water-deficit stress on cotton plants expressing the Bacillus thuringiensis toxin

Bacillus thuringiensis (Bt) crops require a high dosage of Bt toxin to delay development of insect resistance, in particular, when the refuge strategy is applied. This strategy is threatened by plant developmental and environmental factors that might reduce Bt toxin concentration and Bt efficacy in Bt crops. Growth of Bt (Cry1Ac) cotton under prolonged, moderate water deficit as a single stress factor was evaluated. Bt cotton plants were analysed for physiological performance, Bt toxin concentration and Bt efficacy. For performance analysis, leaf and total plant dry weight and leaf area were measured. Bt toxin concentration was determined by an immuno‐assay. Effects of Bt toxin on growth and mortality of African cotton bollworm, Helicoverpa armigera, larvae were measured in different plant organs. Leaves from young plants exposed for 30 days to moderate water deficit had both higher Bt toxin concentrations and were more effective against larvae than leaves, flowers or bolls from mature flowering plants exposed to 60 days of moderate water deficit. Although growth of Bt cotton plants under moderate water‐deficit conditions decreased Bt concentrations in leaves, flowers and bolls, this had no effect on efficacy against first‐instar cotton bollworm larvae. No significant evidence was found that moderate water deficit, as a single stress factor, decreases Bt efficacy in Bt cotton.     

Molecular characterization of a new synthetic cry2ab gene in Nicotiana tabacum

A newly-synthesized cry2Ab gene was characterized in Nicotiana tabacum, before its further transformation in cotton. Synthetic cry2Ab gene was cloned in pGreen0029 and its expression was transiently analyzed at mRNA level through agroinfiltration in tobacco. The mRNA of cry2Ab was detected after 72 h agroinfiltration through PCR using total plant RNA. This construct was then transformed into N. tabacum through Agrobacterium. Insect bioassays were conducted on detached leaves using first instar Spodoptera exigua larvae; after 96 h significant insect mortality was recorded. This newly synthesized gene was effective in controlling S. exigua first instar larvae. It can be used in combinations with other Bt genes like cry1Ac for developing resistance against major insect pests of cotton and further widening the insect control spectrum.     

Transformation and inheritance of Bt genes inGossypium hirsutum

Transgenic plants offer many unique opportunities for managing pest populations. However, the inheritance, integration, and expression of multiple transgenes are prerequisite for maintaining sustainable resistance against insects in crops. We took a gene-pyramiding approach to produce Bt cotton expressing two Bt genes,cry1Ac andcry2A. Using sonication-assistedAgrobacterium-mediated transformation (SAAT), we achieved an efficiency of 6.26%. Putative transgenic plants were confirmed via PCR, Southern hybridization, and western-blotting. Those showing mortality of 75 to 100% for the second instar ofHeliothis armigera (compared with 0% for the control) were considered Bt-positive. Transgenes were segregated according to a 3:1 Mendelian inheritance pattern in the T1 generation forHeliothis resistance. In our insect bioassay, the control plants showed >95% leaf damage, and insects reached the 4^th instar stage of larval growth. In contrast, leaf damage on transgenic plants was limited to only a few bites, and insect mortality was 75 to 100%. ELISA confirmed transgene expression, and Bt protein was detected in leaf tissue. This performance was consistent with that of the parent transgenics. PCR and Southern blots verified integration of thecry1Ac andcry2A genes into the progeny. Therefore, this strategy provides a pathway toward cotton improvement and the development of durable resistance against insect damage.     

Clonostachys rosea BAFC3874 as a Sclerotinia sclerotiorum antagonist: mechanisms involved and potential as a biocontrol agent

Aims: To establish the modes of action of the antagonistic fungal strain Clonostachys rosea BAFC3874 isolated from suppressive soils against Sclerotinia sclerotiorum and to determine its potential as a biocontrol agent. Methods and Results: The antagonistic activity of C. rosea BAFC3874 was determined in vitro by dual cultures. The strain effectively antagonized S. sclerotiorum in pot‐grown lettuce and soybean plants. Antifungal activity assays of C. rosea BAFC3874 grown in culture established that the strain produced antifungal compounds against S. sclerotiorum associated with secondary metabolism. High mycelial growth inhibition coincided with sclerotia production inhibition. The C. rosea strain produced a microheterogeneous mixture of peptides belonging to the peptaibiotic family. Moreover, mycoparasitism activity was observed in the dual culture. Conclusions: Clonostachys rosea strain BAFC3874 was proved to be an effective antagonist against the aggressive soil‐borne pathogen S. sclerotiorum in greenhouse experiments. The main mechanisms involve peptaibiotic metabolite production and mycoparasitism activity. Significance and Impact of the Study: Clonostachys rosea BAFC3874 may be a good fungal biological control agent against S. sclerotiorum. In addition, we were also able to isolate and identify peptaibols, an unusual family of compounds in this genus of fungi.     

Characterization of the <Emphasis Type="Italic">Bacillus thuringiensis</Emphasis> Isolates Virulent Against Rice Leaf Folder, <Emphasis Type="Italic">Cnaphalocrocis medinalis</Emphasis> (Guenee)

Soil samples were collected from different rice fields of Singur, Hooghly, West Bengal, India. Spore forming bacteria were isolated from the soil samples and among them, two isolates (BUSNC25 and BUSNC26) were larvicidal against third, fourth and fifth instar larvae of rice leaf folder, Cnaphalocrocis medinalis. The phenotypic, biochemical characterization and 16S rDNA analysis of the two isolates were done. On the basis of phenotypic, biochemical and phylogenetic analysis, the selected bacterial isolates (BUSNC25 and BUSNC26) were identified as Bacillus thuringiensis. The antibiotic sensitivity tests of these two isolates against selected doses of some standard antibiotics were done. Against the 3rd, 4th and 5th instar larvae of C. medinalis, the LC₅₀ values of BUSNC25 were 2.45 × 10⁴, 1.325 × 10⁴ and 2.35 × 10⁴ cfu/ml and of BUSNC26 were 3.375 × 10⁴, 1.9 × 10⁴ and 3.325 × 10⁴ cfu/ml, respectively.     

Feeding‐induced phenol production in Capsicum annuum L. influences Spodoptera litura F. larval growth and physiology

We studied the role of induced plant phenols as a defense response to insect herbivory. Phenolic compounds were induced in Capsicum annuum L., the source of many culinary peppers, after feeding by different stages of the insect pest, Spodoptera litura F. The phenols were identified and quantified using high performance liquid chromatography (HPLC) and effects produced by these phenols on larval development were studied. Vanillic acid was identified in plants challenged by second, fourth, and fifth instar larvae, but not in plants challenged by third instar nor unchallenged plants. Syringic acid production was induced in chili plants infested with second (0.429 ± 0.003 μg/g fresh weight, fourth (0.396 ± 0.01 μg/g fresh weight), and fifth instar (5.5 ± 0.06 μg/g fresh weight) larvae, compared to untreated plants (0.303 ± 0.01 μg/g fresh weight) plants. Leaves surface treated with the rutin deterred oviposition. Dietary exposure to chlorogenic acid, vanillic acid, syringic acid, sinapic acid, and rutin led to enhanced activities of detoxifying enzymes, β‐glucosidase, carboxyl esterase, glutathione S‐transferase, and glutathione reductase in the midgut tissues of all the larval instars, indicating the toxic nature of these compounds. Protein carbonyl content and acetylcholinesterase activity was analyzed to appreciate the role of induced plant phenols in insect protein oxidation and terminating nerve impulses.     

Control of Bemisia tabaci by entomopathogenic fungi isolated from arid soils in Argentina

Entomopathogenic Hypocreales were isolated from arid soils in Argentina using Tenebrio molitor as bait and tested for their biological performance at 30°C and 45–65% RH. Conidial germination was tested in three vegetable oils (sunflower, olive and maize) at two concentrations (1% and 10%) to evaluate their compatibility for further liquid formulations. According to radial growth and germination results, we selected four isolates to test their pathogenicity against second instar B. tabaci nymphs with the selected oil formulations at 30°C. CEP381 and CEP401 showed the highest radial growth. Isolates CEP381, CEP401, CEP413 and CEP409 (Metarhizium spp.) had similar germination percentages as compared with water control when germinated on either sunflower, olive or maize oils at 10% v/v. The highest mortality of B. tabaci was observed for the isolates CEP381 in sunflower oil and CEP401 in olive oil. Molecular identification of isolates was performed using ITS4–5 primers. All isolates belong to the Metarhizium core group. Tested isolates could grow and infect B. tabaci nymphs at 30°C in some of the vegetable oils as carriers, providing new possibilities for integrated pest management of Bemisia tabaci.     

Organogenesis and plant regeneration in Zephyranthes rosea Lindl.: Histological and chromosomal study

The genus Zephyranthes rosea is a member of the family Amaryllidaceae. The plant is widely cultivated as ornamental. The objective of this study was to optimize an in vitro propagation method for the production of genetically stable Z. rosea plant. The chromosomal status of the regenerated plants was also studied to determine their ploidy levels and to identify the structural and numerical variations, if any. Two explants of Zephyranthes rosea, i.e. bulb scale and flower bud (3–4 mm each), were used and incubated in a culture room at 25 ± 2°C in which two different types of calli were induced from two sources. The MS medium amended with 2,4-dichlorophenoxyacetic acid (2,4-D) (0.5–2.0 mg/l) successfully induced callus from bulb-scale explants (50.25–57.5%). The addition of coconut water (10%) in 2,4-D-added medium further improved the callus induction frequency (68.4%). Bulb-scale calli were found to be highly regenerative while flower-bud calli did not show any organogenetic responses. The use of plant growth regulators, such as naphthaleneacetic acid (NAA) + benzylaminopurine (BAP), was found to be very effective for shoot bud development; maximum shoot number (11.50/callus mass) was observed in NAA (0.5 mg/l) + BAP (1.0 mg/l) added medium. Histological analysis of callus revealed that the origin of the shoot bud was de novo. Rooting frequency (65.25%) and the number of roots (7.5/shoot) were best achieved in indole-3-butyric acid (4.0 mg/l)-amended medium, followed by indole-3-acetic acid (4.0 mg/l). The regenerated Z. rosea plants showed 2n = 24 chromosome numbers.     

Bacillus thuringiensis isolates from soil and diseased insects in Egyptian cotton fields and their activity against lepidopterous insects

The distribution of Bacillus thuringiensis in Egyptian soils of cotton cultivations represented by 11 governorates was studied. This study aimed to isolate indigenous strains that may be potent against some lepidopterous insect pests. Out of 45 isolates, only 10 were effective and they showed high levels of toxicity against the target insects at 500 μg/ml (80% larval mortality or higher). Among these 10 isolates, two isolates were potent against Spodoptera littoralis, two isolates were potent against Helicoverpa (= Heliothis) armigera, seven isolates were potent against Pectinophora gossypiella, but none were potent against Agrotis ypsilon. LC₅₀ and LC₉₀ values and the potency of the B. thuringiensis isolates have been determined. Trials were conducted to isolate B. thuringiensis from diseased lepidopterous insects collected from the same locations, but all isolates showed low potential activity against the target insects. These findings showed promise for the possible use of some indigenous B. thuringiensis strains in the control of lepidopterous pests in Egypt.     

Suppressive soil against Sclerotinia sclerotiorum as a source of potential biocontrol agents: selection and evaluation of Clonostachys rosea BAFC1646

The fungal diversity structures of soils that are suppressive and non-suppressive to Sclerotinia sclerotiorum were characterised and screened for fungal strains antagonistic to the S. sclerotiorum pathogen. Soil suppressiveness was associated with a particular fungal diversity structure. Principal component analysis showed that antagonism by fungal species in suppressive soils was associated with the occurrence of Fusarium oxysporum, Fusarium solani, Talaromyces flavus var. flavus and Clonostachys rosea f. rosea. In particular, C. rosea f. rosea occurred exclusively in suppressive soil samples, suggesting that this morpho-species plays an important role in suppression of S. sclerotiorum diseases. One strain of C. rosea f. rosea (BAFC1646) was selected for further experiments. Dual-culture assays confirmed the antagonistic behaviour of C. rosea f. rosea BAFC1646 against three different S. sclerotiorum strains. Antifungal activity was corroborated by diffusion assays with metabolite extracts. Greenhouse assays with soybean plants showed that the selected C. rosea f. rosea strain reduced the percentage of dead plants when co-inoculated with S. sclerotiorum. In addition, inclusion of C. rosea f. rosea alone increased shoot lengths significantly. In this work, we established the involvement of fungal species in soil suppressiveness and in further assays confirmed that C. rosea f. rosea BAFC1646 exhibits a bioprotective effect against S. sclerotiorum in soybean plants.     

Biology and performance of two indigenous larval parasitoids on Tuta absoluta (Lepidoptera: Gelechiidae) in Sudan

Tuta absoluta is an alien invasive pest in Sudan. Since it was detected in the country, the pest continues to cause major tomato losses. Alarmed by its devastating nature and the speed of spread, tomato growers resorted to the indiscriminate use of broad-spectrum insecticides. Promising indigenous parasitoid(s) may contribute to suppression of this pest. The biology and performance of the native parasitoids, Bracon nigricans Szépligeti (Braconidae) and Dolichogenidea appellator (Telenga) (Braconidae), was therefore evaluated against different immature stages of T. absoluta under laboratory conditions. A significantly higher number of fourth instar larvae was accepted for oviposition by B. nigricans. Similarly, fourth instar larvae yielded a considerably higher number of parasitoid offspring compared to third instar larvae. The male ratio of B. nigricans offspring which emerged from both larval instars was high, 0.96 and 0.66 for third and fourth instar larvae of T. absoluta, respectively. There was, however, no significant difference in the number of males and females that emerged from fourth instar larvae. Conversely, the number of hosts killed by stinging behaviour or host feeding was significantly higher on third instar larvae. B. nigricans potential fecundity varied with the age of the wasp with the lowest fecundity on the day of eclosion. The performance of D. appellator in terms of the total number of offspring produced and female progeny was similar for second and third instar larvae of T. absoluta. The preimaginal developmental time for both parasitoid species did not vary with either host stage or sex of the parasitoid. B. nigricans adult longevity was similar for both sexes, while that of D. appellator significantly varied with females living longer than males. Prospects for conservative biological control of T. absoluta are discussed in the light of the results of this study.     

Water stress and kaolin spray affect herbivorous insects’ success on cotton

Ecological hypotheses of plant–insect herbivore interactions suggest that insects perform better on weakened plants and plants grown under optimal conditions are less damaged. This study tested the hypothesis that the colonization and oviposition rates by pests with different feeding strategies and levels of specialization are affected in different ways by two conditions commonly faced by commercially grown plants–water deficit and application of kaolin sprays, a reducer of abiotic plant stressors. We used four major pests of cotton as insect herbivore models. Three were chewing Lepidoptera: Alabama argillacea (Hüb.), a monophagous pest on cotton; Heliothis virescens (Fabr.), which is polyphagous, but with cotton as a primary host; and Chrysodeixis includens (Walk.), which is polyphagous, with cotton as secondary host. The fourth pest was a sap-sucking species, the polyphagous whitefly Bemisia tabaci (Gen.). In both choice and no-choice trials, the three chewing pests oviposited significantly less upon water-stressed plants; the greatest effect was observed for C. includens (>90 % reduction in oviposition under choice and >58 % under no-choice conditions). In contrast, the sap-sucking B. tabaci exhibited statistically more colonization and oviposition on water-stressed plants. Application of kaolin sprays reduced colonization and oviposition by all herbivore species tested, irrespective of irrigation regime and feeding strategies.     

Factors influencing colonization of cucumber roots by Clonostachys rosea f. catenulata,a biological disease control agent

The biocontrol fungus Clonostachys rosea f. catenulata (Gliocladium catenulatum) strain J1446, commercially available as Prestop® (Verdera Oy, Finland), is an effective antagonist against several root and foliar greenhouse pathogens. The biocontrol agent forms dense networks of hyphae on plant roots, grows internally in root epidermal cells, and produces hydrolytic enzymes, all of which lead to a reduction in pathogen propagules. An understanding of the environmental and host factors that influence root colonization by C. rosea f. catenulata is important to maximize disease control efficacy. Cucumber roots grown in nutrient solution in containers were inoculated with conidia of a GUS-transformed strain of C. rosea f. catenulata. Population levels associated with roots over time were assessed by colony-plate counts, GUS staining and enzymatic assays to determine GUS activity. Variables such as pH, temperature and growing medium were major factors that influenced population levels, while cucumber cultivar, addition of nutrients, and wounding of roots did not appear to significantly affect colonization. Population density of C. rosea f. catenulata on roots was highest when the nutrient solution was maintained at pH 5, 6, or 7, and at temperatures of 18–22°C. Lowest colonization levels were observed on roots of plants grown in potting mix or in field soil. Measurement of GUS activity provided a slightly more accurate assessment of root colonization levels compared to colony-plate counts. These results illustrate the optimal environmental conditions which can ensure maximum root colonization by C. rosea f. catenulata and enhance disease control by the biocontrol agent.     

Bacillus thuringiensis Isolates from Great Nicobar Islands

Bacillus thuringiensis (Bt) strains were isolated from soil samples of Great Nicobar Islands, one of the “hottest biodiversity hotspots,” where no collection has been characterized previously. The 36 new Bt isolates were obtained from 153 samples analyzed by crystal protein production with light/phase-contrast microscopy, determination of cry gene profile by SDS-PAGE, evaluation of toxicity against Coleopteran, and Lepidopteran insect pests, finally cloning and sequencing. Majority of the isolates showed the presence of 66–35 kDa protein bands on SDS-PAGE while the rest showed >130, 130, 73, and 18 kDa bands. The variations in crystal morphology and mass of crystal protein(s) purified from the isolates of Bt revealed genetic and molecular diversity. Based on the toxicity test, 50 % of isolates were toxic to Ash weevils, 16 % isolates were toxic to cotton bollworm, 38 % isolates were toxic both to ash weevil as well as cotton bollworm, while 11 % of the isolates did not exhibit any toxicity. PCR analysis unveiled prepotency of cry1B- and cry8b-like genes in these isolates. This study appoints the first isolation and characterization of local B. thuringiensis isolates in Great Nicobar Islands. Some of these isolates display toxic potential and, therefore, could be adopted for future applications to control some agriculturally important insect pests in the area of integrated pest management for sustainable agriculture.     

Insecticidal efficacy of two isolates of Beauveria bassiana (Bals.) (Vuill.), on the second larval stage of Leptinotarsa decemlineata (Say) (Col.: Chrysomelidae)

Leptinotarsa decemlineata is a serious potato pest throughout the world that has developed resistance to many insecticides. This study investigated the virulence of two indigenous isolates of Beauveria bassiana, consisting of DEBI007 and IR1217C. Five different concentrations of each isolate, 1 × 10⁷, 3.1 × 10⁷, 1 × 10⁸, 3.1 × 10⁸ and 1 × 10⁹ conidia ml^?1, were applied in bioassay using dipping method on second instar larvae. Control insects were treated with distilled water containing 0.01% Tween-80 (Sigma, Germany) as surfactant. The results showed that mortality percentages due to DEBI007 and IR1217C isolates in the highest concentration were 60% and 57.77%, respectively. There was a significant difference between efficacies of different conidial concentrations; therefore, with rise in conidial concentration, the mortality percentage was increased. The DEBI007 isolate that was previously isolated from the soil was more effective than IR1217C isolate. Calculated LC₅₀ for this effective isolate was 1.4 × 10⁷ conidia ml^?1, which, respectively, is the highest concentration.     

Induction of systemic resistance to bacterial blight caused by Xanthomonas campestris pv. malvacearum in cotton by leaf extract from a medicinal plant zimmu (Allium sativum L. × Allium cepa L.)

Abstract

Aqueous extract (10%) from leaves of zimmu (Allium sativum L. × Allium cepa L.) when applied as foliar spray to first and second leaves of cotton plants induced systemic resistance in third and fourth leaves to a challenge infection with Xanthomonas campestris pv. malvacearum and reduced the number of lesions by up to 73% compared with water-treated control plants. The treated leaves exhibited significantly high activity of enzymes phenylalanine ammonia-lyase, peroxidase and polyphenol oxidase along with rapid accumulation of phenolics. The activities of chitinase and β-1,3-glucanase were greatly elevated in treated plants as compared to water-treated controls. An 11-fold increase in chitinase activity was evident 4 d after treatment. Western blot analysis revealed that a chitinase with an apparent molecular weight of 58 kDa that cross-reacted with a barley chitinase antiserum was induced in cotton leaves 3 d after treatment and the maximum induction of this chitinase was detected 4 d after treatment. The present study provides evidence for the induction of biochemical defence mechanisms in cotton leaves after treatment with leaf extract from zimmu.     

Molecular characterization of Inonotus rickii /Ptychogaster cubensis isolates from different geographic provenances

Thirteen isolates of Inonotus rickii/Ptychogaster cubensis, from different geographic provenances, were analyzed by sequencing ITS1, ITS 2 and 5,8S ribosomal RNA region. A phylogenetic tree, also including sequences available in Genbank database, showed that the strains enclosed in this study fall into two well-separated groups, one formed by isolates from Florida (USA) and the other one by isolates from Europe, Argentina and China. Differences were also highlighted on the growth rate of mycelial cultures at different temperatures. In fact, although the tested isolates generally attained the best growth at 30°C, isolates from Europe seem well adapted to higher temperatures and went on growing at 40°C whilst the growth of isolates from Florida significantly decreased at 35°C. Since the teleomorph I. rickii was never detected in Florida, and in this study noticeable differences were detected by analysis of ITS region, the existence of two possible distinct species, not discriminated solely on the basis of morphological characters, could be suggested.