Elicitors directed in vitro growth and production of stevioside and other secondary metabolites in Stevia rebaudiana (Bertoni) Bertoni.

Stevia rebaudiana (stevia) is a plant in the Asteraceae that contains several biologically active compounds including the antidiabetic diterpene glycosides (e.g. stevioside, rebaudioside and dulcoside) that can serve as zero-calorie sugar alternatives. In this study, an elicitation strategy was applied using 5% polyethylene glycol (PEG), sodium chloride (NaCl; 50 and 100 mM) and gibberellic acid (2.0 and 4.0 mg/L GA3) to investigate their effect on shoot morphogenesis, and the production of phenolics, flavonoids, total soluble sugars, proline and stevioside, as well as antioxidant activity, in shoot cultures of S. rebaudiana. Herewith, the media supplemented with 2 mg/L and 4 mg/L GA3 exhibited the highest shooting response (87% and 80%). The augmentation of lower concentrations of GA3 (2 mg/L) in combination with 6-benzylaminopurine (BAP) resulted in the maximum mean shoot length (11.1 cm). The addition of 100 mM NaCl salts to the media led to the highest observed total phenolics content (TPC; 4.11 mg/g-DW compared to the control 0.52 mg/g-DW), total flavonoids content (TFC; 1.26 mg/g-DW) and polyphenolics concentration (5.39 mg/g-DW) in shoots cultured. However, the maximum antioxidant activity (81.8%) was observed in shoots raised in media treated with 50 mM NaCl. The application of 2 mg/L of GA3 resulted in the highest accumulation of proline (0.99 µg/mL) as compared to controls (0.37 µg/mL). Maximum stevioside content (71 µL/mL) was observed in cultures supplemented with 100 mM NaCl and 5% PEG, followed by the 4 mg/L GA3 treatment (70 µL/mL) as compared to control (60 µL/mL). Positive correlation was observed between GA3 and stevioside content. Notably, these two compounds are derived from a shared biochemical pathway. These results suggest that elicitation is an effective option to enhance the accumulation of steviosides and other metabolites and provides the groundwork for future industrial scale production using bioreactors.

Evaluation of Insecticidal Potentials of Five Plant Extracts against the Stored Grain Pest, Callosobruchus maculatus (Coleoptera: Bruchidae).

Plant based insecticides are considered among the most economic and ecofriendly chemicals for the protection of plants and stored grains. The cowpea weevil (Callosbruchus maculatus) causes more than 90% damage to sored grains in three to six months. The current study investigates insecticidal potentials of five selected botanicals: Melia azedarach, Nicotiana rustica, Azadirachta indica, Nicotiana tabacum and Thuja orientalis. They are explored at six different concentrations (0.5, 1.0, 1.5, 2.0, 2.5 and 3.0%) against C. maculatus and compared to effects of distilled water which is used as a control. Toxicities of 3%(V/V) extracts of N. tabacum, N. rustica, A. indica and T. orientalis against C. maculatus were 100%, 86.11%, 80.56% and 72.22%, respectively. Maximum mortality was caused by N. tabacum and N. rustica (100%), followed by A. indica (82%), whereas minimum mortality was observed in T. orientalis (64%) at 2.5%. Several phytochemicals, alkaloids, saponins, diterphenes, phytosterol, flavonoids and phenols were identified in N. tabacum and N. rustica, while few were present in A. indica. Phytosterol was present in greatest abundance. Saponins were only detected in aqueous extracts of N. rustica and N. tabacum. Taken together, these results indicate the utility of N. tabacum, N. rustica and A. indica as potential botanicals to control pest beetle and cowpea weevil.

Biorational Control of Callosobruchus maculatus (Coleoptera: Buchidae) in Stored Grains with Botanical Extracts.

Globally, around 2000 plant species are used against pest control. The utilization of botanicals is considered the most economic and biodegradable methods for the control of stored grains pests. Therefore, the current study was carried out to investigate the repellency potential of five botanicals against Callosbruchus maculatus F. in Haripur, Pakistan. The concentrations of Azadirachta indica L., Nicotiana tabacum L., Melia azedarach L., Nicotiana rustica L., and Thuja orientalis L. were, i.e., 0.5, 1.0, 1.5, 2.0, 2.5, and 3.0% in four replicates to establish contact effects. The data were recorded after 1, 2, 3, 6, 24, 48, 72, and 96 hours. The repellency effect of these plant species against C. maculatus were increased in both the time- and dose-dependent manner, and highest effect was observed at 72 h. In addition, the repellency effect was 91% for A. indica (class: V), 86% M. azedarach, 82%, N. tabacum (class: V), 79% N. rustica (class: IV), and 75% T. orientalis (class: IV) at 3% concentration against C. maculatus. Furthermore, following 96 hours' exposure to treatment the sensitivity response of insects decreases as the time interval increases, i.e., 86% A. indica (class: V) was followed by 71% M. azedarach (class: IV), 65% N. tabacum (class: IV), 61% N. rustica (class: IV), and T. orientalis 57% (class: III) repellency at highest concentration of 3%. The current study concluded that A. indica and M. azedarach can be incorporated for the management of C. maculatus and these plant species might be helpful in the productions of new biopesticides.

Optimization of Biostimulants Application for Phenology and Quality of Oats

Abstract In modern agriculture the use of biostimulants not only reduces the cost of production but also increase yield and quality of crops without harming environment. The response of fodder oat to biostimulants was evaluated in a pot experiment using four biostimulants at different concentrations i.e Moringa leaf extract at ratio of 1:10, 1:20, 1:30, 1:40, 1:50, Sea weed extract at 1%, 2%, 3%, 4%, 5%, Thiourea at 100 ppm, 200 ppm, 300 ppm, 400 ppm, 500 ppm and Chitosan at 25 ppm, 50 ppm, 100 ppm, 125 ppm, 150 ppm. Oat crop was treated with biostimulants using foliar application, at two growth stages i.e tillering and booting. Data on various agronomic and quality parameters were recorded and analyzed. Results showed that biostimulants significantly affected phenology and quality of oat. The highest green fodder yield (872.32 g) was observed in the pots treated with 2%Solution of seaweed extract similarly highest dry matter yield was also highest at 170.85 g Interestingly crude protein was highest for thiourea at 400 ppm. In each biostimulant following doses stood out both in terms of yield and quality. In case of moringa leaf extract the ratio of 1:40with 900 g per pot yield and 9.43 % protein, for sea weed extract 2% was better than other doses with 981 g yield and 9.17% protein, for thiourea the 400 ppm was better than other concentrations as it shows highest yield (872.32 g) and crude protein (9.52%) and for chitosan 100 ppm was comparatively better in performance with 964.98 g yield per pot and 9.35% CP. It is concluded from our study that biostimulants improve fodder oat yield and quality.

The genus Cryptogonus Mulsant (Coleoptera: Coccinellidae) from Pakistan, with description of a new species and a new record.

The new species of a ladybird beetle, Cryptogonus bhalwalnensis Azad Wang sp. nov. is described from Pakistan. Cryptogonus nepalensis bhutanensis Bielawski is recorded for the first time from Pakistan. Both species of Cryptogonus from Pakistan are diagnosed and illustrated; a distribution map is also presented.

Storage Stability of Kinnow Fruit (Citrus reticulata) as Affected by CMC and Guar Gum-Based Silver Nanoparticle Coatings.

The influence of carboxy methyl cellulose (CMC) and guargum-based coatings containing silver nanoparticles was studied on the postharvest storage stability of the kinnow mandarin (Citrus reticulata cv. Blanco) for a period of 120 days (85%-90% relative humidity) at 4 °C and 10 °C. Physicochemical and microbiological qualities were monitored after every 15 days of storage. Overall results revealed an increase in total soluble solid (TSS), total sugars, reducing sugars and weight loss but this increase was comparatively less significant in coated fruits stored at 4 °C. Ascorbic acid, total phenolics, and antioxidant activity was significantly enhanced in coated fruits stored at 4 °C. Titratable acidity significantly decreased during storage except for coated kinnow stored at 4 °C. In control samples stored at 10 °C, high intensity of fruit rotting and no chilling injury was observed. Total aerobic psychrotrophic bacteria and yeast and molds were noticed in all treatments during storage but the growth was not significant in coated fruits at 4 °C. Kinnow fruit can be kept in good quality after coating for four months at 4 °C and for 2 months at 10 °C.

Development of inexpensive and globally available larval diet for rearing Anopheles stephensi (Diptera: Culicidae) mosquitoes.

BACKGROUND: Success of sterile insect technique (SIT) is dependent upon the mass rearing and release of quality insects, the production of which is directly related to the suitability of the diet ingredients used. Commercial diets used for small-scale culture of mosquitoes are expensive and thus not feasible for mass production. METHODS: A series of low cost globally available diet ingredients including, wheat, rice, corn, chickpeas, and beans along with liver, were provided to 4 h larvae (L1) of Anopheles stephensi (Liston) to see their effect on fitness parameters including larval duration, percent emergence, survival, adult wing size and female fecundity. Different quantities of the candidate diet ingredients were then mixed together to work out a combination diet with a balanced nutritive value that can be used for efficient rearing of the mosquito larvae at relatively lower costs. RESULTS: Fastest larval and pupal development and highest survival rates were recorded using a combination diet of bean, corn, wheat, chickpea, rice, and bovine liver at 5 mg/day. The diet is easy to prepare, and much cheaper than the diets reported earlier. The estimated cost of the reported diet is 14.7 US$/ 1.3 kg for rearing one million larvae. CONCLUSIONS: A combination diet with ingredients from cereals and legumes mixed with liver is a low cost balanced larval diet with the potential for use in both small scale laboratory rearing and mass production of Anopheles in SIT control programs.