Green peach aphid, Myzus persicae (Hemiptera: Aphididae), is an economically important pest of crops within more than 40 plant families all over the world. This study encompasses in-vitro pathogenicity of two strains of Beauveria bassiana (BB-72 and BB-252) and one strain of Lecanicillium lecanii (V-4) against green peach aphid (M. persicae). Using a leaf-dip method, three different bioassays were conducted comprised of filtrates and conidial concentrations of BB-72, BB-252 and V-4 fungal strains and their binary combinations. Infiltrate bioassays, 2 mL fungal filtrate of each strain was used. In conidial bioassays, three different concentrations (i.e., 1 × 10⁶, 1 × 10⁷ and 1 × 10⁸ conidia mL⁻¹) of each fungal strain were used, while in binary combination bioassays, LC₅₀ and LC₃₃ of these fungal strains were evaluated. According to the results, maximum pathogenicity against test aphids was observed at 10th day of all treatments. Both strains of B. bassiana (BB-72 and BB-252) exhibited higher mortality of aphids (up to 95 and 91%, respectively) than L. lecanii (V-4) strain (87%) in all three types of bioassays. Moreover, binary combinations of BB-72 and BB-252 strains showed significantly higher aphid mortality (94%) than other combinations. Reduced mortality in case of fungal combinations of V-4 strain might be due to the antagonistic effect of L. lecanii strain along with both strains of B. bassiana. Nevertheless, evaluation of combined pathogenicity of the fungal strains constitutes the novelty of this in-vitro study which revealed that both strains of B. bassiana (i.e., BB-72 and BB-252) are mutually compatible and can be utilized together as new biocontrol tools against destructive insect pests such as M. persicae.

Elicitors are biofactors that induce resistance in plants against different insect pests. This in vitro study evaluated the impact of a novel elicitor protein PeBC1, extracted from a necrotrophic fungus Botrytis cinerea, on the development and fecundity parameters of green peach aphid (Myzus persicae) on common beans (Phaseolus vulgaris L.). Three different concentrations of PeBC1 elicitor (i.e., 33.56, 25.43, 19.33 µg mL⁻¹) were applied at three different temperature regimes (i.e., 18, 21, and 25 °C). Elicitor treatments were applied topically on the bean plants at 3-leaf stage and newly emerged (0–6 h old) apterous adult aphids were exposed to these treated leaves. In addition to the biological parameters of aphids, the relative expression levels of key genes associated with jasmonic acid (JA) and salicylic acid (SA) plant defense pathways were also determined through RT-qPCR. Results of bioassays revealed that the application of PeBC1 elicitor protein exhibited pronounced and significant (p < 0.05) sub-lethal effects on green peach aphids. The fecundity was reduced and the nymphal development time was prolonged by different concentrations of PeBC1 elicitor and temperature regimes. Gene expression studies showed that the exogenous application of PeBC1 induced a significant upregulation of the expression levels of JA and SA pathway-associated genes in bean plants. As compared to control, elicitor-treated plants exhibited an induced resistance against aphids. Our findings suggest the potential use of PeBC1 elicitor protein in future bio-intensive management strategies against sap-sucking insect pests such as green peach aphids.

This study reports the characterization of protein elicitor PeBb1 derived from entomopathogenic fungus Beauveria bassiana ARSEF-2860 strain and its putative role in induced systemic resistance in Brassica rapa ssp. pekinensis against green peach aphid Myzus persicae. The sequence of purified elicitor protein was matched with the genomic sequence of a hypothetical protein BBA_10269 from B. bassiana ARSEF-2860 (GenBank Accession No. XP_008603588.1). The protein-encoding gene PeBb1 contained 534 bp cDNA encoding a polypeptide of 177 amino acids with a molecular mass of 19 kDa. The recombinant elicitor protein was expressed in Escherichia coli using pET-28a (+) expression vector and induced necrosis in the leaves of tobacco. The effects of elicitor protein on aphid M. persicae was determined by applying three different concentrations of PeBb1 (i.e., 26, 35, 53 μM) on B. rapa plants at 4-leaf stage and the treated plants were exposed to newly emerged (0–6 h old) apterous adult aphids. Bioassay results showed significant (p < 0.05) sub-lethal effects of the exogenous application of PeBb1 elicitor on M. persicae. Moreover, the RT-qPCR gene expression analyses showed a significant up-regulation of most of the key genes linked to ethylene (ET)- and jasmonic acid (JA)-associated plant defense pathways in elicitor-treated plants. These results not only recommend the putative utilization of PeBb1 elicitor protein in future biological pest control strategies against phloem-feeding insect pests such as M. persicae, but also help in better comprehension of the mechanisms through which beneficial fungi trigger the induced plant resistance.

Bacterial microbe-associated molecular patterns (MAMPs) play an important role in innate plant immunity. This in vitro study evaluated the putative role of protein elicitor PeBA1 derived from Bacillus amyloliquefaciens NC6 strain in eliciting induced resistance type responses in common bean (Phaseolus vulgaris) plants against green peach aphid Myzus persicae. Nymphal developmental time of aphids was significantly prolonged and the fecundity was significantly reduced by different concentrations of PeBA1 elicitor (i.e. 40.51, 24.91 and 16.38 µg mL^-1) applied at three different temperature regimes (i.e. 21, 27 and 30 °C). Moreover, foliar application of PeBA1 elicitor protein strongly up-regulated the expression levels of salicylic acid (SA) pathway-associated genes, while the expression levels of jasmonic acid (JA) pathway-associated genes exhibited a moderate induction. Quantification by LC/MSMS revealed a linear increase of both SA and JA plant defense hormones along with the time of exposure. Our findings suggest that the bacterial elicitor protein PeBA1 could be used as an effective biological pest management tool against phloem-feeding insect pests such as green peach aphids M. persicae.

Bell pepper (Capsicum annuum L.) is one of the extensively cultivated vegetable crop in Punjab, Pakistan. During two years of field surveys, February-November 2016-17, damping off and blight symptoms were observed. Average seedling mortality was recorded as 18.7% while yield loss due to blight was estimated 32 to 41% at mature stages. Maximum blight infection was recorded from the areas frequently flooded with canal irrigation system. At early stages, lesions were noticed on stem portions at soil line level while at crop maturity stages blight symptoms were noted. Leaves were blanched and wilted while fruits were covered with white mold. Masses of sporangia were evident on and inside the infected fruits under humid conditions. A total of twelve isolates were recovered from infected root, stem and fruit portions on rye agar media (Caten and Jinks, 1968) incubated at 25oC under fluorescent light. Papillated sporangia were averaged 42 ± 2.6 X 27 ± 1.7 μm in size (range 27 - 52 × 23 - 36 μm). Oospores were produced on 20% V8 agar and were spherical 22 ± 1.4 μm in diameter (range 14 to 27 μm) while average pedicels length was recorded as 58 ± 12.5 μm (range 13 to 120 μm). These observations were similar to those described for P. capsici (Cocoa, 1988). DNA was extracted using Cetyl Trimethylammonium Bromide (CTAB) method and the internal transcribed spacer regions (ITS1 and ITS2) were amplified by polymerase chain reaction (White et al., 1990). The amplicons were purified and sequenced in both directions (GenBank Accession No. MF322868 and MF322869). BLAST analysis revealed these isolates showed 99% identity with ITS sequences of Phytophthora capsici (KM369964 and KU518782). Pathogenicity assay was performed on healthy bell pepper seedlings with five repeats. Soil was flooded with 20ml sporangial suspension (1 x 103 sporangia/ml) in pots containing seedlings while 5ml suspension was sprayed until run off on mature plants (Hyder et al., 2018). A set of uninoculated seedlings was used as control. Pots were kept in dew chamber for 10-20 days at 25±2 oC. Seedling mortality was observed five days after inoculation while at later stage plants develop brown-to-black stem lesions with white mycelial growth on leaves. These symptoms were identical to the P. capsici infections in field. Consistent re-isolations of P. capsici confirm its association with the disease. To our knowledge, this is the first report of Phytophthora blight on bell pepper from Pakistan

Nile Tilapia cultivation with biofloc in the coastal area of Lake Batur as an alternative to using floating net cages has not been well developed. The main objective of this research is to analyze the efficiency of tilapia cultivation with biofloc in the coastal area of Lake Batur. This research is essential for developing tilapia cultivation with biofloc and limiting cultivation using floating net cages. The study was conducted in Songan B Village, Kintamani Bangli, involving a sample of farmers cultivating tilapia with biofloc. Data collection uses a questionnaire survey method to analyze the efficiency of tilapia cultivation businesses using analysis of the R/C Ratio. The research found that tilapia cultivation with biofloc was efficient with an R/C value = 1.02, a low survival rate of 66.69%, and a relatively large feed conversion ratio of 1: 1.52.