Regeneration in Jatropha curcas: Factors affecting the efficiency of in vitro regeneration

Factors influencing in vitro regeneration through direct shoot bud induction from hypocotyl explants of Jatropha curcas were studied in the present investigation. Regeneration in J. curcas was found to be genotype dependent and out of four toxic and one non-toxic genotype studied, non-toxic was least responsive. The best results irrespective of genotype were obtained on the medium containing 0.5 mg L⁻¹ TDZ (Thidiazuron) and in vitro hypocotyl explants were observed to have higher regeneration efficiency as compared to ex vitro explant in both toxic and non-toxic genotypes. Adventitious shoot buds could be induced from the distal end of explants in all the genotypes. The number of shoot buds formed and not the number of explants responding to TDZ treatment were significantly affected by the position of the explant on the seedling axis. Explants from younger seedlings (≤15 days) were still juvenile and formed callus easily, whereas the regeneration response declined with increase in age of seedlings after 30 days. Transient reduction of Ca²⁺ concentrations to 0.22 g L⁻¹ in the germination medium increased the number of responding explants.Induced shoot buds, upon transfer to MS medium containing 2 mg L⁻¹ Kn (Kinetin) and 1 mg L⁻¹ BAP (6-benzylamino purine) elongated. These elongated shoots were further proliferated on MS medium supplemented with 1.5 mg L⁻¹ IAA (indole-3-acetic acid) and 0.5 mg L⁻¹ BAP and 3.01-3.91 cm elongation was achieved after 6 weeks. No genotype specific variance in shoot elongation was observed among the toxic genotypes except the CSMCRI-JC2, which showed reduced response. And for proliferation among the toxic genotypes, CSMCRI-JC4 showed highest number of shoots formed. Among the rest, no significant differences were observed. The elongated shoot could be rooted by pulse treatment on half-strength MS medium supplemented with 2% sucrose, 3 mg L⁻¹ IBA (indole-3-butyric acid), 1 mg L⁻¹ IAA, 1 mg L⁻¹ NAA (α-naphthalene acetic acid) and subsequent transfer on 0.25 mg L⁻¹ activated charcoal medium. The rooted plants could be established in soil with more than 90% success. No significant differences were observed in rooting of shoots in the different toxic genotypes. However, rooting response was reduced in non-toxic genotype as compared to toxic genotypes.     

In vitro regeneration from petiole explants of non-toxic Jatropha curcas

Jatropha curcas, a multipurpose shrub has acquired significant economic potential as biodiesel plant. The seeds or pressed cake is toxic due to the presence of toxic substances and is not useful as food/fodder despite having the best protein composition. A simple, efficient, and reproducible method for plant regeneration through direct organogenesis from petiole explants of non-toxic J. curcas was developed using Murashige and Skoog (MS) medium supplemented with different concentrations of thidiazuron (TDZ). The best induction of shoot buds (57.61%), and number of shoot buds (4.98) per explant were obtained when in vitro petiole explants were placed horizontally on MS medium supplemented with 2.27 μM TDZ. The Induced shoot buds were transferred to MS medium containing 10 μM kinetin (Kn), 4.5 μM 6-benzyl aminopurine (BA), and 5.5 μM α-naphthaleneacetic acid (NAA) for shoot proliferation and subsequent elongation was achieved on MS medium supplemented with 2.25 μM BA and 8.5 μM IAA. The elongated shoots could be rooted on half-strength MS medium with 15 μM IBA, 11.4 μM IAA and 5.5 μM NAA with more than 90% survival rate.     

Assessment of dermal wound healing and in vitro antioxidant properties of Avena sativa L.

Avena sativa L. (Poaceae) has been reported to have traditional utilization against skin diseases and inflammation. Therefore, in this study, the n-hexane, ethyl acetate, ethanol, and water extracts of A. sativa were investigated for their wound healing and antioxidant activities. Total phenol and flavonoid contents of the extracts were established spectrophotometrically. For the wound healing activity, linear incision and circular excision models on rats and mice were evaluated with a standard ointment Madecassol^®. Antioxidant activity was evaluated by 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging, ferrous ion-chelating, and ferric-reducing antioxidant power (FRAP) assays. Significant wound healing activity was observed with the ointment formulation of the ethanol extract at 1% concentration. The histopathological examination results also supported the outcome of both linear incision and circular excision wound models. All of the extracts exerted low antioxidant activity in the applied assays. The present study provides a scientific evidence for the traditional usage of A. sativa in the management of wound healing.     

High level of resistance to Sclerotinia sclerotiorum in introgression lines derived from hybridization between wild crucifers and the crop Brassica species B. napus and B. juncea

Sclerotinia rot caused by the fungus Sclerotinia sclerotiorum is one of the most serious and damaging diseases of oilseed rape and there is keen worldwide interest to identify Brassica genotypes with resistance to this pathogen. Complete resistance against this pathogen has not been reported in the field, with only partial resistance being observed in some Brassica genotypes. Introgression lines were developed following hybridization of three wild crucifers (viz. Erucastrum cardaminoides, Diplotaxis tenuisiliqua and E. abyssinicum) with B. napus or B. juncea. Their resistance responses were characterized by using a stem inoculation test. Seed of 54 lines of B. napus and B. juncea obtained from Australia, India and China through an Australian Centre for International Agricultural Research (ACIAR) collaboration programme were used as susceptible check comparisons. Introgression lines derived from E. cardaminoides, D. tenuisiliqua and E. abyssinicum had much higher levels (P < 0.001) of resistance compared with the ACIAR germplasm. Median values of stem lesion length of introgression lines derived from the wild species were 1.2, 1.7 and 2.0 cm, respectively, as compared with the ACIAR germplasm where the median value for stem lesion length was 8.7 cm. This is the first report of high levels of resistance against S. sclerotiorum in introgression lines derived from E. cardaminoides, D. tenuisiliqua and E. abyssinicum. The novel sources of resistance identified in this study are a highly valuable resource that can be used in oilseed Brassica breeding programmes to enhance resistance in future B. napus and B. juncea cultivars against Sclerotinia stem rot.     

Large scale in vitro propagation of Stevia rebaudiana (bert) for commercial application: Pharmaceutically important and antidiabetic medicinal herb

Stevia rebaudiana is a valuable medicinal plant species and it is being used for the treatment of diabetes. Currently, there is a high demand for raw material of this medicinal herb due to ever increasing diabetes disorder among the population. In order to meet the increased demand an efficient in vitro propagation of S. rebaudiana was established. Nodal explants collected from the field were cultured on MS basal medium fortified with different concentrations of BAP (0.5-3.0 mg/l) and KIN (0.5-3.0 mg/l) individually for shoot bud induction. In vitro derived nodal buds were cultured on MS medium supplemented with different concentrations (0.5-3.0 mg/l) of BAP and KIN for multiple shoot bud regeneration. In the second experiment, in vitro derived buds were placed on MS medium supplemented with different concentrations of BAP (0.5-3.0 mg/l) in combination with 0.5 mg/l IAA or IBA or NAA for shoot bud multiplication. The highest frequency (94.50%) of multiple shoot regeneration with maximum number of shoots (15.69 shoots/explant) was noticed on MS medium supplemented with 1.0 mg/l BAP. For large scale plant production, in vitro derived nodal bud explants were cultured on MS medium fortified with 1.0 mg/l BAP, in which about 123 shoots/explant were obtained after three subcultures on the same media composition. Elongated shoots (>2 cm) dissected out from the in vitro proliferated shoot clumps were cultured on half-strength MS medium containing different concentrations of NAA (0.1-0.5 mg/l) and/or MS medium fortified with various concentrations (0.5-2.0 mg/l) of auxins (NAA, IAA and IBA) for root induction. Highest frequency of rooting (96%) was noticed on half-strength MS medium augmented with 0.4 mg/l NAA. The rooted plantlets were successfully transferred into plastic cups containing sand and soil in the ratio of 1:2 and subsequently established in the greenhouse. The present in vitro propagation protocol would facilitate an alternative method for rapid and large-scale production of this important antidiabetic medicinal plant.     

In vitro plant regeneration from decapitated embryonic axes of Clitoria ternatea L.—An important medicinal plant

In vitro clonal propagation of Clitoria ternatea has been achieved by employing decapitated embryonic axes (DEAs) explants. The explants induced multiple shoots on cytokinin-containing medium. Several cytokinins [6-benzylaminopurine (BAP), 6-furfuryl aminopurine (KIN) and thidiazuron (TDZ)] were assayed. The best response was achieved with 2 mg l⁻¹ BAP in which 100% of cultures produced 6.0 ± 0.14 shoots per explant. MS + 1 mg l⁻¹ gibberellic acid (GA₃) was the most suitable for shoot elongation. Regenerated shoots were rooted in half-strength Murashige and Skoog (MS) medium with 0.2 mg l⁻¹ indole-3-butyric acid (IBA). Plantlets were successfully acclimatized and established in soil, and they were morphologically indistinguishable from the source plant. The plantlets attained maturity and flowered normally. The efficient regeneration protocol reported here provides an important method of micropropagation of this plant. Furthermore, this protocol may be used for genetic transformation of this valuable medicinal plant for its further improvement.     

Maize genotype susceptibility to Rhizoctonia solani and its effect on sugar beet crop rotations

Root and crown rot is the major soil-borne fungal disease in sugar beet. In Europe, the disease is mainly caused by the anastomosis group (AG) 2-2IIIB of the basidiomycete Rhizoctonia solani (Kühn). No chemical fungicide to control the disease has been registered in Europe. Therefore, agronomic measures must be optimized to keep the disease severity below an economic damage threshold and to minimize white sugar yield losses. R. solani AG 2-2IIIB infects many other crops besides sugar beet, including maize, where it causes root rot. Sugar beet and maize are frequently grown in the same crop rotation. The proportion of cultivated maize in several European sugar beet growing areas is expected to rise due to a projected increase in demand for renewable resources over the next few years. Although the susceptibility to and tolerance of the disease varies among cultivars in both crops, little is known about the effects of cultivar susceptibility in the pre-crop on a subsequent susceptible crop. The cultivation of R. solani-resistant maize genotypes in rotation with resistant sugar beet might therefore be a useful tool in an integrated control strategy against R. solani, eliminating the need to restrict the desired crop rotation for phytosanitary reasons. A crop rotation experiment with artificially inoculated R. solani was conducted in the field to investigate the pre-crop effects of maize cultivars which differed in their susceptibility to R. solani on a susceptible sugar beet cultivar. We hypothesized that the maize genotype would influence the inoculum potential and performance of a susceptible sugar beet genotype grown after a maize pre-crop, and that this would correlate with the susceptibility of the maize genotype. The results demonstrate that the susceptibility of maize genotypes is consistent over a period of years and that cultivated maize genotypes influenced the inoculum potential measured as disease severity in sugar beet. However, disease severity in sugar beet did not correlate with the disease susceptibility of the genotype of the maize pre-crop. Possible reasons for this missing relationship might be differences in the quality of maize residues for the saprophytic survival of the pathogen or a genotype-specific alteration of the antagonistic microbial community. However, our findings showed that in the presence of maize- and sugar beet-pathogenic R. solani, the most favourable maize cultivar for a crop rotation cannot be determined solely on the basis of its resistance level against Rhizoctonia root rot.     

Sensitivity of Sclerotinia sclerotiorum to fludioxonil: In vitro determination of baseline sensitivity and resistance risk

This work determined the sensitivity of field populations of Sclerotinia sclerotiorum (Lib.) de Bary before exposure to the fungicide fludioxonil (= baseline sensitivity) and assessed the risk of fludioxonil resistance. The mean EC₅₀ (Effective Concentration) and Minimum inhibitory concentration (MIC) values for fludioxonil based on inhibition of mycelial growth of 120 wild-type isolates were 0.015 ± 0.005 μg/ml and <0.05 μg/ml, respectively. Positive cross-resistance was not detected between fludioxonil and benzimidazole fungicides but was detected between fludioxonil and dicarboximide fungicides which are considered as high resistance risk fungicides by FRAC, even though these fungicides have different molecular structures. By growing wild-type isolates on potato dextrose agar (PDA) containing sublethal concentrations of the fungicide, we obtained four fludioxonil-resistant mutants with resistance factors (EC₅₀ resistant/EC₅₀ sensitive phenotypes) >2000. The laboratory fludioxonil mutants were less fitter than their parental isolates in terms of mycelial radial growth, pathogenicity and sclerotial production. Moreover, on PDA amended with NaCl, the laboratory fludioxonil mutants grew more slowly than their fludioxonil-sensitive parents, especially at lower concentrations of NaCl. According to the fitness of mutants and the cross-resistance between fludioxonil and dicarboximide fungicides, phenylpyrroles can be considered to pose a moderate resistance risk. In a field trial, fludioxonil provided greater control (over 90% disease control) of S. sclerotiorum than iprodione.     

Feeding behavior of a potential insect pest, Lygus hesperus, on four new industrial crops for the arid southwestern USA

Camelina (Camelina sativa), guayule (Parthenium argentatum), lesquerella (Physaria fendleri), and vernonia (Centrapalus pauciflorus [formerly Vernonia galamensis]) are either under limited commercial production or being developed for production in the southwestern USA. Insect pests are a potential economic threat to all these new crops, with Lygus hesperus, the western tarnished plant bug, among the most prominent due to its regional abundance and propensity to feed on reproductive plant tissue. The objectives of this study were to establish baseline data on the feeding behavior and potential impact of L. hesperus on camelina, guayule, lesquerella and vernonia. Behavioral observations of adult females and males, and nymphs of this insect were made in the laboratory. Insects spent ≈35% of their time either probing (=tasting) or feeding on various reproductive and vegetative tissues of guayule, lesquerella or vernonia, but only 20% on camelina. When insects did probe and feed they preferred reproductive tissues, primarily flowers and siliques/achenes, and there were differences in these behaviors relative to crop but not generally to insect stage or sex. Insects probed and fed more on flower tissue of guayule and vernonia compared with camelina and lesquerella, and more on siliques of lesquerella compared with achenes of vernonia. When probing and feeding on vegetative tissue, there was generally a preference for stems compared with leaves in all crops except guayule. Results show that L. hesperus will readily feed on the economically important tissues of all crops, and although research has shown that this feeding did not consistently affect lesquerella yield, further work will be needed to determine if such feeding poses a risk to commercial production of camelina, guayule or vernonia.     

In vitro activity of eighteen essential oils and some major components against common postharvest fungal pathogens of fruit

The development of natural crop protective products as alternatives to synthetic fungicides is currently in the spotlight. In vitro experiments are valuable precursors to more costly in vivo trials, allowing the identification of effective essential oils and establishing the concentrations required for inhibition of a specific, or spectrum of decay pathogens. In this study, the antifungal properties of eighteen essential oils were evaluated in vitro by addition to the fungal growth medium of five pathogens (Lasiodiplodia theobromae, Colletotrichum gloeosporioides, Alternaria citrii, Botrytis cinerea and Penicillium digitatum) isolated from mango, avocado, citrus, grapes and cactus pear. The inhibitory properties of some of the major compounds of the oils, identified and quantified by gas chromatography-mass spectroscopy and gas chromatography-flame ionization detection were also determined. Most of the oils were selected on the basis of commercial availability and for containing a predominant compound. Visual inspection of fungal growth was done and the lowest concentration where fungal growth was completely inhibited on all replicates was recorded. Thyme oil proved to be the most effective inhibitor, totally inhibiting all of the pathogens tested at concentrations of 1000 μl/l and lower, with the exception of a resistant Penicillium strain. Cinnamon oil, rich in eugenol (81.2%), demonstrated good fungicide potential, while the carvone-rich oils displayed promising activity against the citrus pathogens. Oils characterized by high concentrations of S-carvone were less effective than those containing the R-enantiomer. Essential oil of Lippia citriodora was active against all of the pathogens, excluding L. theobromae from avocado. These essential oils, applied alone or in combination, are good candidates for further in vivo testing and for investigations concerning their modes of action.     

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.     

Azoxystrobin resistance of French Mycosphaerella graminicola strains assessed by four in vitro bioassays and by screening of G143A substitution

We tested here five distinct in vitro screening methods assessing Mycosphaerella graminicola resistance to strobilurin fungicides: (i) spotting on agar medium, (ii) spore germination on agar medium, (iii) growth in microplates without and (iv) with the addition of Alamar blue and (v) screening of the cytochrome b substitution conferring resistance G143A. The assays were performed by assessing resistance to azoxystrobin of 32 French M. graminicola strains and the two reference strains IPO323 and IPO94269 from the Netherlands. The microplate Alamar blue assay displayed high standard deviations from all growth averages, hence reporting the strong lack of reliability of this method. As expected, the two reference strains were found sensitive with all other methods. In agreement with the disruptive resistance of M. graminicola to single-site fungicides, the half maximal inhibitory concentration (IC₅₀) values of sensitive and resistant strains displayed a bimodal distribution pattern with each type of assay. All strains carrying G143 or A143 alleles were found sensitive or resistant respectively with each bioassay. Although the absolute IC₅₀ value for each strain varied among assays, the ranking of strains according to their IC₅₀ values was overall conserved with all assays. Finally, the frequency of strobilurin resistant haplotypes within France was investigated by screening G143A in 82 strains isolated in 2005 year from 12 localities. Results showed a marked gradual decrease in resistance distribution from 70% in northern to 30% and 0% in central and southern France, respectively. The accuracy of each method as well as the widespread and incidence of strobilurin resistance within the French population of M. graminicola have been discussed.     

Effect of herbicide and cover crop on weed control in no-tillage jack-o-lantern pumpkin (Cucurbita pepo L.) production

Field experiments were conducted to evaluate cover crop (none, winter wheat, or winter rye) and pre-emergence (PRE) applications of clomazone plus ethalfluralin alone or with halosulfuron [PRE or post-emergence (POST)] for smooth crabgrass [Digitaria ischaemum (Schreb. ex Schweig) Schreb. ex Muhl.] and redroot pigweed (Amaranthus retroflexus L.) control in no-tillage ‘Aspen’ jack-o-lantern pumpkin (Cucurbita pepo L.). At pumpkin harvest, cover crops had reduced smooth crabgrass density, but not redroot pigweed. Although PRE or POST applications of halosulfuron alone were more effective at reducing redroot pigweed density than clomazone plus ethalfluralin PRE, the combination of halosulfuron plus clomazone and ethalfluralin PRE reduced redroot pigweed density to the greatest extent. Pumpkin yields were not affected by cover crop, although average pumpkin sizes were greater with the inclusion of cover crop residues. Pumpkin fruit sizes and yields were the greatest with clomazone and ethalfluralin PRE in combination with halosulfuron applied PRE or POST. Overall, cover crop had relatively little influence on pumpkin yields compared with herbicide treatments. The addition of halosulfuron to clomazone and ethalfluralin provided greater broadleaf weed control resulting in greater jack-o-lantern pumpkin yields.     

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.     

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.     

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.     

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.     

Mycoparasitic Trichoderma viride as a biocontrol agent against Fusarium oxysporum f. sp. adzuki and Pythium arrhenomanes and as a growth promoter of soybean

Trichoderma viride was proved as an effective biocontrol agent against two fungal pathogens, Fusarium oxysporum f. sp. adzuki and Pythium arrhenomanes, infecting soybean. During an in vitro biocontrol test, Trichoderma showed mycoparasitism and destructive control against the tested fungal pathogens. Both the pathogens significantly influence the germination and P. arrhenomanes had a severe effect (only 5% germination). The root system of the soybean plant was poorly developed due to the infection and it exerted a negative influence on the nodulation and further growth phases of the plant. During pot assay along with biocontrol activity, Trichoderma showed growth promoting action on the soybean plant. Trichoderma enhanced growth of shoot and root systems and fruit yield after 12 weeks of growth. Pythium and Fusarium infected plants treated with Trichoderma had ∼194% and 141% more height than pathogens alone. The fruit yield treated with Trichoderma was ∼66 per plant whereas the yield was only 41 for a control plant. The plants infected with Pythium and Fusarium and treated with Trichoderma had fruit yields of 43 and 53 respectively and those were 5 and 1.6 times higher than plants infected with pathogens.     

NaCl plays a key role for in vitro micropropagation of Salicornia brachiata, an extreme halophyte

A simple and rapid method for micropropagation of succulent, salt accumulator and extreme halophyte Salicornia brachiata has been established for the first time using shoot tips and nodes. Individually, BA showed significant response compared to Kn and in combinations, improved shoot proliferation was observed with BA + NAA than BA + 2,4-D, however no significant response was observed with BA + IAA. Percentage of shoot response significantly increased with NaCl treatment in the combination of BA + NAA while BA + 2,4-D + NaCl combination showed reduced shoot proliferation followed by demises of most of cultures. Efficient shoot proliferation was observed with combinations BA (8.9 μM) + NAA (5.37 μM) + NaCl (500 mM) and BA (13.3 μM) + NAA (5.37 μM) + NaCl (250 mM) indicating that NaCl is required for the micropropagation. The developed method will facilitate functional analysis of novel salt responsive gene(s) isolated from S. brachiata and propagation of industrially important elite accessions.