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Plants have been used since ancient times as an important source of biologically active substances. Specific activities of these plant extracts are generally linked to the presence of secondary metabolites together with their phenolic contents. Present study aimed at investigating the total phenolic and flavonoid contents, and antioxidant activity of selected plants from five different families. The total phenolic content was measured using Folin-Ciocalteu assay and total flavonoid content by aluminum chloride colorimetric method. The antioxidant capacity was estimated by phosphomolybdinium assay. Our findings indicates that total phenolic content for methanolic extracts ranged from 27.07 to 59.11 mg GAE/g DW, and total flavonoid content ranged from 38.37 to 124.23 mg QE/g DW, with an antioxidant activity ranging from 55.82 to 129.06 mg AAE/g DE. Following trend was shown in the assessment of total phenolic and flavonoid contents: Rhazya stricta>Cicer arietinum>Solanum melongena>Solanum surattense>Solanum nigrum>Withania sominifera>Sisymbrium irio>Withania coagulans>Raphanus sativus>Fagonia indica>Brassica napus. While the antioxidant capacity followed the trend: Cicer arietinum>Solanum nigrum>Withania coagulans>Rhazya stricta>Raphanus sativus>Solanum melongena>Withania sominifera>Solanum surratense>Fagonia indica>Brassica napus>Sisymbrium irio. It is also seen that both wild and cultivated plants have higher medicinal value, which can be linked to the phenolic and flavonoid content, and antioxidant potential. Findings of the study revealed that wild plants possess higher phenolic content compared to cultivated plants, whereas cultivated plants had higher antioxidant activity.

The yield of crops is largely affected by different types of biotic stresses. To minimize the damage, crop plants adapted themselves to overcome the stress conditions through gene expression reprogramming at transcriptional and post-transcriptional levels. With a better knowledge of plants' responses in adverse environments, new methodologies and strategies have been applied to develop better stress-tolerant plants. In this manner, small RNAs (micro RNA and small-interfering RNA) are reported to play a central role to combat biotic stresses in plants. Depending upon the stress stimuli, these small RNAs can up or down regulate the genes expression, that indicate their potential role in overcoming the stress. These stress-induced small RNAs may reduce the expression of the target gene(s) that might negatively influence plants' response to the adverse conditions. Contrariwise, miRNA, a class of small RNA, can downregulate its expression to upregulate the expression of the target gene(s), which might positively aid to the stress adaptation. Along with this, benefits of RNA interference (RNAi) have also been stated in functional genomic research on insects, fungi and plant pathogens. RNAi is involved in the safe transport of dsRNA to the targeted mRNA(s) in the biotic stress-causing agents (for example fungi and insects) and saves the plant from damage, which is a safer approach compared to use of chemical pesticides. The current review summarizes the role of small RNAs and the use of RNAi to save the plants from biotic stress conditions.

Different parts ofMethanolic, acetone, and n-hexane extracts of selected species were prepared, and ten secondary metabolites were examined using standard protocols. The antioxidant activity was performed using threeTheThe

Ajuga bracteosa Wall. ex Benth is an endangered medicinal herb used against different ailments in folklore medicines. Here, we aimed to create a new insight to the fundamental mechanisms of genetic transformation in the ethnomedicinal usage of this plant. We transformed the plant with rol genes of Agrobacterium rhizogenes and raised the regenerants from the hairy roots. The transgenic regenerants were screened for in vitro antioxidant activities, a range of in vivo assays, and linked the activities with elemental analysis, polyphenol content and different phytochemicals found through HPLC. Among 18 polyphenolic standards, kaempferol was found most abundant in all transgenic lines (up to 101.26 ± 6 µg/mg). Furthermore, among all tested plant extracts, transgenic line 3 (ABRL3) showed maximum phenolics (13.39 ± 2µg GAE/mg) and flavonoids content (4.75 ± 0.16 µg QE/mg). ABRL3 also demonstrated potent total antioxidant capacity (8.16 ± 1 µg AAE/mg), total reducing power, (6.60 ± 1.17 µg AAE/mg), DPPH activity (IC50 = 59.5 ± 0.8µg/mL), hydroxyl ion scavenging (IC50 = 122.5 ± 0.90 µg/mL), and iron chelating power (IC50 = 154.8 ± 2 µg/mL) among all plants. Transformed plant extracts also produced significant analgesic, anti-inflammatory, anticoagulant, and antidepressant properties in in vivo mice model as compared to control untransformed plant material. Additionally, no abnormal behavior or lethality was observed in any animal tested. In conclusion, transgenic regenerants of A. bracteosa pose better pharmacological properties under the effect of rol genes as compared to wild type plants.

Infusions, decoctions and tinctures were prepared from flowers of Butea monosperma (Lam.) Taub. and Sesbania grandiflora (L.) Poiret and evaluated for in vitro inhibition of enzymes implicated on the onset of neurological diseases (acetylcholinesterase: AChE and butyrylcholinesterase: BuChE), diabetes (alpha-glucosidase and alpha-amylase), obesity (lipase) and skin hyperpigmentation (tyrosinase). Extracts were also appraised for radical scavenging activity (RSA) on 2,2-diphenyl-1-picrylhydrazyl (DPPH) and 2,2'-azino-bis (3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) radicals, and for metal chelating activity on copper and iron ions. Samples were evaluated for their total contents in different phenolics groups by spectrophotometric methods, for phenolic profile by high performance liquid chromatography e diode array detection (HPLC-DAD) and for mineral contents by microwave plasma-atomic emission spectrometry (MP-AE). Regarding B. monosperma, the tincture allowed for a moderate inhibition of AChE, the decoction was able to inhibit alpha-glucosidase and no activity was observed towards BuChE, alpha-amylase or lipase. All extracts had a low or moderate inhibition towards tyrosinase, and significant RSA and metal chelating potential. As for S. grandiflora, only the decoction inhibited AChE, none of the extracts was able to inhibit BuChE, all samples inhibited alpha-glucosidase and infusions and decoctions had similar inhibitory properties towards alpha-amylase. None of the extracts was active against lipase, but all were able to inhibit tyrosinase. Extracts had also significant RSA, moderate copper chelation and decoctions had the capacity to chelate iron. The most abundant macroelements in both species were potassium and calcium, while iron was the prevalent microelement, especially in B. monosperma. Both species had significant levels of phenolic compounds, and the main components in decoctions and infusions of B. monosperma were syringic and salicylic acids, while the major compound identified in tinctures was the flavonoid luteolin-7-O-glucoside. In S. grandiflora the most abundant were chlorogenic and neochlorogenic acids and catechin hydrate. Molecular docking studies on the most abundant molecules in S. grandiflora, (+)-catechin, chlorogenic acid and neochlorogenic acid, indicate that these compounds are able to dock to alpha-glucosidase in a similar manner than acarbose. Our results suggest that flowers of both species are a promising source of high value-added compounds with enzyme inhibitory and antioxidant properties. (c) 2018 SAAB. Published by Elsevier B.V. All rights reserved. Foundation for Science and Technology (FCT)Portuguese Foundation for Science and Technology Portuguese National Budget [CCMAR/Multi/04326] FCT Investigator Programme [IF/00049/2012] FCTPortuguese Foundation for Science and Technology [SFRH/BD/94407/2013, SFRH/BD/116604/2016]

The world is experiencing a cancer epidemic and an increase in the prevalence of the disease. Cancer remains a major killer, accounting for more than half a million deaths annually. There is a wide range of natural products that have the potential to treat this disease. One of these products is artemisinin; a natural product from Artemisia plant. The Nobel Prize for Medicine was awarded in 2015 for the discovery of artemisinin in recognition of the drug's efficacy. Artemisinin produces highly reactive free radicals by the breakdown of two oxygen atoms that kill cancerous cells. These cells sequester iron and accumulate as much as 1000 times in comparison with normal cells. Generally, chemotherapy is toxic to both cancerous cells and normal cells, while no significant cytotoxicity from artemisinin to normal cells has been found in more than 4000 case studies, which makes it far different than conventional chemotherapy. The pleiotropic response of artemisinin in cancer cells is responsible for growth inhibition by multiple ways including inhibition of angiogenesis, apoptosis, cell cycle arrest, disruption of cell migration, and modulation of nuclear receptor responsiveness. It is very encouraging that artemisinin and its derivatives are anticipated to be a novel class of broad-spectrum antitumor agents based on efficacy and safety. This review aims to highlight these achievements and propose potential strategies to develop artemisinin and its derivatives as a new class of cancer therapeutic agents.

Ajuga bracteosa a highly demanding pharmaceutical plant, is getting endangered in several regions of the world due to ruthless and destructive harvesting. Research efforts need to be done for its sustainable production and utilization. In this study, effects of Thidiazuron (TDZ) in comparison to conventional cytokinins were tested in vitro on shoot morphogenesis, antioxidant system and biosynthesis of volatile organic compounds in A. bracteosa. Thidiazuron alone acted as the best growth regulator for direct shoot morphogenesis in explants by promoting highest induction of shoots (88%) in stem explants cultivated on MS (Murashige and Skoog) medium supplemented with 1.2 ppm (TDZ). At lower doses (0.5 ppm), TDZ resulted in highest multiplication (18 shoots) and elongation (5.8 cm) of shoots. Further, shoot cultures raised in vitro at TDZ (1.2 ppm) displayed higher activities of superoxide dismutase (SOD: 4.2 U/mg protein), peroxidase (POD: 2.8 U/mg protein) and ascorbate peroxidase (APx: 1.8 U/mg protein), respectively. Maximum total phenolic content (TPC; 4.5 mg GAE/g DW) and total flavonoid content (TFC; 3.5 mg QE/g DW), respectively were detected in shoot cultures grown over MS medium combined with 1.2 ppm TDZ plus 2.0 ppm benzylaminopurine (BA). Analysis through gas chromatography-mass spectrometry (GC–MS) showed, biosynthesis of 34 volatile compounds in the in vitro raised shoots. Wherein, TDZ (1.2 ppm) resulted in production of substantial amounts of bioactive monoterpene hydrocarbons including α-pinene (5.3%), camphene (4.45%), limonene (3.4%), 1,8–cineole (14.3%), α-thujone (9.4%) and oxygenated monoterpenes such as camphor (12.2%), borneol (11.4%) and nerol (9.2%). Thus, besides providing an ex-situ conservation strategy for a high valued medicinal herb, this study has the potential for commercial production of important bioactive volatile compounds.

Ajuga bracteosa is a medicinally important plant globally used in the folk medicine against many serious ailments. In the present study, effects of two significant elicitors, methyl jasmonate (Me-J) and phenyl acetic acid (PAA) were studied on growth parameters, secondary metabolites production, and antioxidant potential in adventitious root suspension cultures of A. bracteosa. The results showed a substantial increase in biomass accumulation, exhibiting longer log phases of cultures growth in response to elicitor treatments, in comparison to control. Maximum dry biomass formation (8.88 DW g/L) was recorded on 32nd day in log phase of culture when 0.6 mg/L Me-J was applied; however, PAA at 1.2 mg/L produced maximum biomass (8.24 DW g/L) on day 40 of culture. Furthermore, we observed the elicitors-induced enhancement in phenolic content (total phenolic content), flavonoid content (total flavonoid content) and antioxidant activity (free radical scavenging activity) in root suspension cultures of A. bracteosa. Application of 0.6 mg/L and 1.2 mg/L of Me-J, root cultures accumulated higher TPC levels (3.6 mg GAE/g DW) and (3.7 mg GAE/g DW) in the log phase and stationary phase, respectively, while 2.5 mg/L Me-J produced lower levels (1.4 mg GAE/g DW) in stationary phase of growth stages. Moreover, TFC and FRSA values were found in correspondence to TPC values in the respective growth phases at the similar elicitor treatment. Thus, a feasible protocol for establishment of adventitious roots in A. bracteosa was developed and enhancement in biomass and metabolite content in adventitious root was promoted through elicitation.

Ajuga bracteosa is a valuable medicinal plant producing many important compounds, including withanolides, phytoecdysteroids, neo-clerodane-di and triterpenes, iridoid glycosides, flavonoids, etc. These compounds possess a broad spectrum of biological, pharmacological and medicinal properties, but their yield is very low in wild-type plants, and chemical synthesis is not viable. Metabolic engineering strategies offer a promising solution for the bulk production of these natural products. However, these strategies usually require the establishment of an efficient genetic transformation method, which to date has not been reported for A. bracteosa . Therefore, the current study was conducted by employing Agrobacterium tumefaciens C58C1 harboring the binary plasmid p35SGUSint with GUS as the reporter gene and the NTPII gene as the selectable marker. The influential factors, e.g. explant type, optical density of agrobacterial culture, inoculation period, co-cultivation duration and acetosyringone concentration, were systematically investigated and the optimal transformation conditions were established by 72 independent transformation experiments. Putative transformants were selected on kanamycin 100 mg/L. MS medium supplemented with 3.6 mg/L BA resulted in maximum regeneration frequency of the transformed explants. The expression of the GUS gene was confirmed by histochemistry and polymerase chain reaction. We established that nodal explants of A. bracteosa precultured for 3 days, inoculated with a culture of A . tumefaciens with an OD of 1.0 for 20 min and co-cultivated for 2 days in MS medium with 200 μM acetosyringone at media pH 5.8 showed 100% transformation induction. This is the first report of a successful A. tumefaciens -mediated transformation of A. bracteosa .

Artemisinin and its analogues are naturally occurring most effective antimalarial secondary metabolites. These compounds also possess activity against various types of cancer cells, schistosomiasis, and some viral diseases. Artemisinin and its derivatives (A&D) are found in very low amounts in the only natural source i.e. Artemisia plant. To meet the global needs, plant sources have been exploited for the enhanced production of these natural products because their chemical synthesis is not profitable. The generally adopted approaches include non-transgenic (tissue and cell cultures) and transgenic together with the cell, tissue, and whole transgenic plant cultures. The genes targeted for the overproduction of A&D include the biosynthetic pathway genes, trichome development genes and rol genes, etc. Artemisinin is naturally produced in trichomes of leaves. At the same time, transgenic hairy roots are considered a good source to harvest artemisinin. However, the absence of trichomes in hairy roots suggests that artemisinin biosynthesis is not limited to trichomes. Moreover, the expression of the gene involved in trichome development and sesquiterpenoid biosynthesis (TFAR1) in transgenic and non-transgenic roots provokes researchers to look for new insight of artemisinin biosynthesis. Here we discuss and review precisely the various biotechnological approaches for the enhanced biosynthesis of A&D. Graphical Abstract

Phytoecdysteroids are secondary metabolites biosynthesized by plants as a defense strategy against phytophagous insects. These are also used in certain medical preparations to reduce depression and prevent infections. We studied the effect of rol genes on phytoecdysteroid biosynthesis in Ajuga bracteosa by transformation through Agrobacterium tumefaciens strain GV3101 harboring pPCV002-ABC. Transformation was confirmed by PCR. Among seven independently generated transgenic lines, a significant increase of phytoecdysteroids was observed in lines 3 and 4 (6728 and 6759 μg g−1 total phytoecdysteroids, respectively) that was 14.5-fold higher than the control plants. Both these lines showed relatively high expression of the rolC gene verified by semi-quantitative RT-PCR and densitometric analysis. We also obtained transgenic hairy root lines of A. bracteosa by transformation with A. rhizogenes strains LBA-9402, A4 and ARqua1. Semi-quantitative RT-PCR and densitometric molecular imaging of 9 transgenic root lines obtained from LBA 9402 revealed a high rolC expression. Transgenic hairy root lines also exhibited phytoecdysteroid production, particularly lines A4-2 and 9402-01 that showed a yield of 4449 and 4123 μg g−1 dry weight, respectively. Complete transgenic plants developed from hairy roots had more phytoecdysteroid content than the parent hairy root lines, suggesting the presence of a possible sink (leaves). Considering the ecdysteroid negative feedback inhibition, we hypothesize that the unavailability of a suitable sink prevents further biosynthesis of phytoecdysteroids in hairy roots. Moreover, sengosterone was not detected in untransformed plants, pPCV002-ABC-generated transgenic plants or untransformed roots, but its presence in some high ecdysteroid-producing hairy root lines suggests its de novo biosynthesis in roots.

Ajuga genus is among one of the more than 250 genera of Labiatae, cosmopolitan in distribution, and comprises of more than 70 species with the remarkable therapeutic importance. Many species of Ajuga including A. bracteosa, A. reptans, A. Chamaepitys, etc. have been used in the traditional system of medicine and are also in use for making formulations in modern medicines. Ajuga species offer anticancer, antibacterial, antifungal, antidiabetic, anabolic, antileishmanial, anti-inflammatory, hepatoprotective, immunomodulatory, antimalarial, astringent, anthelmintic, and diuretic properties and are used in the treatment of rheumatism, palsy, and gout. They hold a large number of secondary metabolites which are active principles to combat the foresaid diseases including phytoecdysteroids, withanolides, iridoid glycosides, neo-clerodane di- and triterpenoids, sterols, and a large range of flavonoid and phenolic compounds. Some of the species of Ajuga are genetically modified for some of these compounds including A. bracteosa, A. reptans, A. multiflora, etc. Latest development made in the exploration of these compounds is yet dealing with the transformation of rol genes and some stress and feeding experiments. We precisely discuss here the details of biotechnological progress that has been made in Ajuga species so far.

The medicinal plant Ajuga bracteosa is a rich source of biologically active and metabolism-enhancing phytoecdysteroids. Transgenic hairy roots from A. bracteosa were obtained by infection with Agrobacterium rhizogenes strains A4, LBA-9402 and ARqua1. The 59 selectively established and rolC-positive hairy root lines increased in size up to 6.6-fold (L42) after one month of in vitro culturing and produced a phytoecdysteroid content ranging from 69.3 to 4449 μg g−1 (L3 and A2, respectively). The clones of these transgenic roots were maintained for successive subcultures on hormone-free medium to obtain a stabilized morphology. Hairy roots displayed four different morphologies: typical hairy root (THR) (59%), callus-like (CM) (17%), thick (TK) (14%) and thin (TN) (10%). The growth rate of the transgenic hairy root lines varied according to their morphology, with CM showing the highest rate (3.93-fold per month). However, THR exhibited the highest phytoecdysteroid content (1538.5 μg g−1). Considering the hairy root morphology in relation to the pRi TR-DNA genes, 100% of the hairy root lines harbored the mas1 and ags genes and 90% of CM lines the aux1 gene. Strikingly, the clones with TK morphology were all positive for the TL- and TR-DNA genes. In contrast, 0% of TN clones harbored the aux1 and ags genes, and only 16.7% the mas1 gene. All CM and TK hairy roots were positive for the ags gene, while mas1 was present in all hairy root types except TN. As expected, all the root lines considered in this work were positive for the rol C gene. Eleven hairy root lines displaying a high growth rate and phytoecdysteroid content were elicited with methyl jasmonate (MeJ) and coronatine (Cor). MeJ doubled the phytoecdysteroid content after 14 days of elicitation (8356 μg g−1 in L2) compared with unelicited control hairy roots, and in in vitro-grown untransformed roots the increase was 5.6-fold.

To elucidate how biosynthesis of plant metabolites is affected by temperature, metabolite profiles from in vitro regenerated plants raised under different temperature regimes of 10, 15 °C, 20 °C, 25 °C and 30 °C were obtained using electrospray ionization mass spectrometry (ESI-MS), and principal component analysis (PCA) was carried out to identify key metabolites. Several bin masses were detected by PCA loading scatter plots which separated the samples. In-house bin program selectively manifested the putative known metabolites depending on % total ions count and intensity of selected bins in the plant samples. Total phenolic and flavonoid content were harvested to highest levels (12.9 mg GAE/g DW and 9.3 mg QE/g DW), respectively, at 15 °C. Besides, pinoresinol (lignan), some of the vital amino acids such as serine, methionine, histidine and glutamine were found to be at higher amount in plants raised at 15 °C. Significant phenylpropanoids like cinnamic acid, caffeic acid and quercitol were detected at a higher concentration in plants raised at 15 °C as compared to other treatments. However, phosphoenolpyruvate, and oxalosuccinate (intermediates of the pentose phosphate pathway) were accumulated the most in plants raised at 30 °C and they were detected with lowest values at 10 °C. Glucose and deoxy-xylose 5 phosphate (intermediates of TCA cycle) were found in higher amounts at temperature treatments of 15 and 25 °C, respectively. We conclude that a low-temperature treatment (15 °C) results in a stress-induced accumulation of a variety of pharmacologically important secondary metabolites.

Centella asiatica (L.) Urban plants have been used since ancient times for their medicinal properties, and their extracts have proven antioxidant, wound healing, sedative, and neuroprotective activities, among others. The natural compounds responsible for C. asiatica bioactivity are triterpene saponins formed from the dammarene branch of the triterpene biosynthetic pathway, collectively known as centellosides, with madecassoside and asiaticoside and their aglycones, madecassic acid and asiatic acid being the most important. Several biotechnological approaches have been developed for the bioproduction of centellosides, based on cell, hairy root, and in vitro plant cultures. This review summarizes the main therapeutic properties of these compounds, as well as their biosynthetic pathways, referring to genetic studies that have identified genes involved in their formation. The biotechnological production of centellosides from a small scale to bioreactor level is also covered. Finally, we summarize the most effective strategies for increasing centelloside yield, including recent transcriptomic, proteomic, and metabolomic studies that have gained new insights into the centelloside biosynthetic pathway and its control.

Lettuce is an important edible crop which possesses various medicinal properties. In this study Lactuca sativa L. (cv Grand Rapids) was transformed by Agrobacterium-mediated transformation with rol C gene. Transgene integration and expression was confirmed through PCR and semiquantitative RT-PCR. The transformed extracts were evaluated for their in vitro antioxidant and in vivo analgesic, anti-inflammatory and antidepressant activities in rats. The transformed plants showed 53-98 % increase in total phenolic and 45-58 % increase in total flavonoid contents compared with untransformed plants. Results of total reducing power and total antioxidant capacity exhibited 90-118 and 61-75 % increase in transformed plants, respectively. In contrast to control, DPPH, lipid peroxidation and DNA protection assay showed up to 37, 20 and 50 % enhancement in transformed plants, respectively. The extracts showed similar but significant enhancement behavior in hot plate analgesic and carrageenan-induced hind paw edema test. The transformed extracts showed 72.1 and 78.5 % increase for analgesic and anti-inflammatory activities, respectively. The transformants of rol C gene exhibited prominent antidepressant activity with 64-73 % increase compared with untransformed plants. In conclusion, the present work suggests that transformation with rol C gene can be used to generate lettuce with enhanced medicinally important properties, such as antioxidant, analgesic, anti-inflammatory and antidepressant potential.

Background Ajuga bracteosa has been extensively used traditionally for the treatment of a variety of diseases. The aim of the study was to scientifically validate the wide-scale exploitation of A. bracteosa in folk medicine various in vitro and in vivo assays. Moreover, these activities were related to the intrinsic biologically active phytoecdysteroids of A. bracteosa. Methods Aerial and root parts of A. bracteosa were first extracted separately with chloroform (AbCA and AbCR) and the residue was again extracted with methanol (AbMA and AbMR). Total flavonoid and phenolic contents were assayed as quercetin (QE) and gallic acid equivalents (GAE), respectively. These extracts were analyzed for in vitro antioxidant assessment including DPPH and H2O2 (% inhibition of free radicals), and reducing power and phosphomolybdenum methods (ascorbic acid equivalents AAE mg/g DW). Further, these extracts were assayed in vivo in separate groups of Sprague–Dawley rats for carrageenan induced rat paw edema inhibition, hotplate antinociception, forced swim antidepression and anticoagulation. Dose of each crude extract and standard drug given to rats was 200 mg/Kg- and 10 mg/10 mL/Kg body weight respectively. Plant extracts and standard drugs were administered orally, 60 min prior to the conduction of assays. Moreover, biologically active phytoecdysteroids were screened in A. bracteosa with the help of RP-HPLC. Results AbMA represented highest values of flavonoids (QE 1.98 % DW) and phenolic contents (GAE 5.94 % DW), significantly scavenged DPPH radicles (IC50 36.9) and reduced ferric ions with 718.4 mg ascorbic acid equivalent/g (AAE). Highest total antioxidant capacity was expressed by AbMR (927 mg AAE) with an IC50 value 19.1 μg/mL. The extracts which were found potent anti-oxidants, were also good at in vivo activities. AbMA significantly reduced edema in all the three hours of treatment (67.9, 70.3 and 74.3 %). AbMA also showed maximum nociceptor suppression in analgesic assay by delaying the time to start licking of paws in rats (57.7 ± 4.9 s). In addition, maximum anti-coagulation was also exhibited by AbMA (89.3 s), while all extracts were found strong antidepressants (≤15.66 s immobility time). Screening of biologically active phytoecdysteroids revealed the presence of 20-hydroxyecdysone (20-HE), makisterone (MKA), cyasterone (CYP) and ajujalactone (AJL). Total phytoecdysteroid content found in A. bracteosa was 1232.5 μg/g DW and 20-HE was most abundant (1232.5 μg/g DW) as compared to other phytoecdysteroids. Conclusion Based on the tested in vitro and in vivo activities, AbMA was found to be a promising bioactive extract. These activities can be attributed to the intrinsic polyphenols and phytoecdysteroids contents of A. bracteosa.

In the study, the residual effect of farmyard manure (FYM) given to previous wheat was evaluated on the next maize crop. A three-crop based experiment maize (control)-wheat-maize from summer 2007 (Kharif) to summer 2008 (Kharif) was performed. Standard dose of FYM was given to the field of intermediate wheat crop. Different NP doses were applied in the four treatments. Residual effect of FYM was estimated out on seven parameters viz; plant height (cm), ear height (cm), cob length (cm), grain row/cob, grains/row, 1000 grains weight (gm) and grain yield/plot (kg). All the parameters exhibited significant results after FYM application except grains/row. Except the ear length, replications remained non-significant. Plant height, cob length and grain yield/plot exhibited fairly significant results with different treatments, while other four parameters remained non-significant. No interaction found between treatments, and before and after FYM application except in 1000 grains weight. Overall 21.37% increase in grain yield/plot was observed. We suggest the application of N 90 kg ha-1 + P 60 kg ha-1 (T2) and N 120 kg ha-1 + P 80kg ha-1 (T3) with standard dose of FYM (6000 kg ha-1) on the previous crop to get better yield of upcoming maize crop in arid regions of Pakistan.

The excessive attention in the use of plants as medicine is credited to the occurrence of active principles whose pharmacological activities have been investigated. Due to the limited production and very less quantity of these important metabolites present in the plant cells, their genetic engineering and increased in vitro production is the point of focus for many years and can be achieved by in vitro transformation and propagation of desired plant. In the current study, we report transformation protocol for Artemisia carvifolia Buch with Agrobacterium tumefaciens C58C1 harboring β-glucuronidase as reporter and neomycin phosphotransferase as selectable marker gene. We have optimized simple regeneration conditions after transformation involving two different types of explants (leaf and stem) on different media formulations for direct organogenesis and best regeneration response. MS media with 2.5 g/L benzylaminopurine (BAP), 0.25 g/L naphthalene acetic acid (NAA), giving maximum number of shoots was selected. Rooting was obtained on ½ MS medium supplemented with NAA (0.1 mg/L). Transient expression of gus reporter gene was observed in the leaf and stem explants after 2 days of bacterial infection. Artemisia carvifolia Buch can be successfully transformed with Agrobacterium tumefaciens strain C58C1 by using controlled in vitro regeneration conditions. Findings of the current study would be useful for micro propagation and genetic transformation of Artemisia carvifolia Buch in future.

From ancient times, plants are being used in treatment of various diseases. Many of today’s drugs have been derived from plant sources. This research work reveals the indigenous medicinal knowledge of important medicinal plants from district Kotli Azad Jammu and Kashmir (AJK), Pakistan. A total 25 common medicinal plants belonging to the 14 families were reported. Their medicinal and other botanically important uses are described by conducting a meeting and interviews from a total of 137 local inhabitants including 73 males, 47 females and 17 Hakims (herbal specialists). Primary source of indigenous medicines were herbs (56%), shrubs (28%) and trees (16%). Herbal preparations were made by the different plant parts. Most common plant part used to make the herbal preparation was leaf (39%) followed by the root (19%), whole plant (12%), seed (9%), bark (7%), fruit (7%), flower (5%) and tuber (2%). Key words: Ethnobotany, Kotli, Azad Jammu and Kashmir, indigenous knowledge.