Commenced in January 2007
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Edition: International
Paper Count: 22

Search results for: Murraya koenigii

22 Regeneration of Plantlets via Direct Somatic Embryogenesis from Different Explants of Murraya koenigii

Authors: Nisha Khatik, Ramesh Joshi


An in vitro plant regeneration system was developed via direct somatic embryogenesis from different seedling explants of an important medicinal plant Murraya koenigii (L) Spreng. Cotyledons (COT), Hypocotyle (HYP)(10 to 15 mm) and Root (RT) segments (10 to 20 mm) were excised from 60 days old seedlings as explants. The somatic embryos induction was achieved on MS basal medium augmented with different concentrations of BAP 1.33 to 8.40 µM and TDZ 1.08 to 9.82 µM. The globular embryos originated from cut ends and entire surface of the root, hypocotyle explants and margins of cotyledons within 30-40days. The percentage of somatic embryos induction per explant was significantly higher in HYP explants (94.21±5.77%) in the MS basal medium supplemented with 6.20 µM BAP and 8.64 µM TDZ. The highest rate of conversion of torpedo, heart and cotyledonary stages from globular stage was obtained in MS medium supplemented with 8.64 µM TDZ. The matured somatic embryos were transferred to the MS basal medium without PGRs. Highest 88% of the matured embryos were germinated on transfer to the PGR free medium where they grew for a further 3-4 weeks. Out of seventy six hardened plants seventy (92%) plantlets were found healthy under field conditions.

Keywords: Murraya koenigii, somatic embryogenesis, thidiazuron, regeneration, rutaceae

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21 Assesment of Genetic Fidelity of Micro-Clones of an Aromatic Medicinal Plant Murraya koenigii (L.) Spreng

Authors: Ramesh Joshi, Nisha Khatik


Murraya koenigii (L.) Spreng locally known as “Curry patta” or “Meetha neem” belonging to the family Rutaceae that grows wildly in Southern Asia. Its aromatic leaves are commonly used as the raw material for traditional medicinal formulations in India. The leaves contain essential oil and also used as a condiment. Several monomeric and binary carbazol alkaloids present in the various plant parts. These alkaloids have been reported to possess anti-microbial, mosquitocidal, topo-isomerase inhibition and antioxidant properties. Some of the alkaloids reported in this plant have showed anti carcinogenic and anti-diabetic properties. The conventional method of propagation of this tree is limited to seeds only, which retain their viability for only a short period. Hence, a biotechnological approach might have an advantage edging over traditional breeding as well as the genetic improvement of M. koenigii within a short period. The development of a reproducible regeneration protocol is the prerequisite for ex situ conservation and micropropagation. An efficient protocol for high frequency regeneration of in vitro plants of Murraya koenigii via different explants such as- nodal segments, intermodal segments, leaf, root segments, hypocotyle, cotyledons and cotyledonary node explants is described. In the present investigation, assessment of clonal fidelity in the micropropagated plantlets of Murraya koenigii was attempted using RAPD and ISSR markers at different pathways of plant tissue culture technique. About 20 ISSR and 40 RAPD primers were used for all the samples. Genomic DNA was extracted by CTAB method. ISSR primer were found to be more suitable as compared to RAPD for the analysis of clonal fidelity of M. koenigii. The amplifications however, were finally performed using RAPD, ISSR markers owing to their better performance in terms of generation of amplification products. In RAPD primer maximum 75% polymorphism was recorded in OPU-2 series which exhibited out of 04 scorable bands, three bands were polymorphic with a band range of size 600-1500 bp. In ISSR primers the UBC 857 showed 50% polymorphism with 02 band were polymorphic of band range size between 400-1000 bp.

Keywords: genetic fidelity, Murraya koenigii, aromatic plants, ISSR primers

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20 In vitro and in vivo Antiangiogenic Activity of Girinimbine Isolated from Murraya koenigii

Authors: Venoos Iman, Suzita Mohd Noor, Syam Mohan, Mohamad Ibrahim Noordin


Girinimbine, a carbazole alkaloid was isolated from the stem bark and root of Murraya koenigii and its structure and purity was identified by HPLC and LC-MS. Here we report that Girinimbine strongly inhibit angiogenesis activity both in vitro and in vivo. MTT result showed that girinimbine inhibits cell proliferation of the HUVECS cell line in vitro. Result of endothelial cell invasion, migration, tube formation and wound healing assays also demonstrated significant time and does dependent inhibition by girinimbine. Moreover, girinibine mediates its anti-angiogenic activity through up- and down-regulation of angiogenic and anti-aniogenic proteins. Furthermore, anti-angiogenic potential of girinimbine was evidenced in vivo on zebrafish model. Girinimbine inhibited neo-vessels formation in zebrafish embryos during 24 hours exposure time. Together, these results demonstrated for the first time that girinimbine could effectively suppress angiogenesis and strongly suggest that it might be a novel angiogenesis inhibitor.

Keywords: anti-angiogenic, carbazole alkaloid, girinimbine, zebrafish

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19 Gastro-Protective Actions of Melatonin and Murraya koenigii Leaf Extract Combination in Piroxicam Treated Male Wistar Rats

Authors: Syed Benazir Firdaus, Debosree Ghosh, Aindrila Chattyopadhyay, Kuladip Jana, Debasish Bandyopadhyay


Gastro-toxic effect of piroxicam, a classical non-steroidal anti-inflammatory drug (NSAID), has restricted its use in arthritis and similar diseases. The present study aims to find if a combination of melatonin and Murraya koenigii leaf extract therapy can protect against piroxicam induced ulcerative damage in rats. For this study, rats were divided into four groups namely control group where rats were orally administered distilled water, only combination treated group, piroxicam treated group and combination pre-administered piroxicam treated group. Each group of rats consisted of six animals. Melatonin at a dose of 20mg/kg body weight and antioxidant rich Murraya koenigii leaf extract at a dose of 50 mg /kg body weight were successively administered at 30 minutes interval one hour before oral administration of piroxicam at a dose of 30 mg/kg body weight to Wistar rats in the combination pre-administered piroxicam treated group. The rats of the animal group which was only combination treated were administered both the drugs respectively without piroxicam treatment whereas the piroxicam treated animal group was administered only piroxicam at 30mg/kg body weight without any pre-treatment with the combination. Macroscopic examination along with histo-pathological study of gastric tissue using haemotoxylin-eosin staining and alcian blue dye staining showed protection of the gastric mucosa in the combination pre-administered piroxicam treated group. Determination of adherent mucus content biochemically and collagen content through Image J analysis of picro-sirius stained sections of rat gastric tissue also revealed protective effects of the combination in piroxicam mediated toxicity. Gelatinolytic activity of piroxicam was significantly reduced by pre-administration of the drugs which was well exhibited by the gelatin zymography study of the rat gastric tissue. Mean ulcer index determined from macroscopic study of rat stomach reduced to a minimum (0±0.00; Mean ± Standard error of mean and number of animals in the group=6) indicating the absence of ulcer spots on pre-treatment of rats with the combination. Gastro-friendly prostaglandin (PGE2) which otherwise gets depleted on piroxicam treatment was also well protected when the combination was pre-administered in the rats prior to piroxicam treatment. The requirement of the individual drugs in low doses in this combinatorial therapeutic approach will possibly minimize the cost of therapy as well as it will eliminate the possibility of any pro-oxidant side effects on the use of high doses of antioxidants. Beneficial activity of this combination therapy in the rat model raises the possibility that similar protective actions might be also observed if it is adopted by patients consuming NSAIDs like piroxicam. However, the introduction of any such therapeutic approach is subject to future studies in human.

Keywords: gastro-protective action, melatonin, Murraya koenigii leaf extract, piroxicam

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18 Antibacterial Activity of Rosmarinus officinalis (Rosemary) and Murraya koenigii (Curry Leaves) against Multidrug Resistant S. aureus and Coagulase Negative Staphylococcus Species

Authors: Asma Naim, Warda Mushtaq


Staphylococcus species are the most versatile and adaptive organism. They are widespread and naturally found on the skin, mucosa and nose in humans. Among these, Staphylococcus aureus is the most important species. These organisms act as opportunistic pathogens and can infect various organs of the host, causing minor skin infection to severe toxin mediated diseases, and life threatening nosocomial infections. Staphylococcus aureus has acquired resistance against β-lactam antibiotics by the production of β-lactamase, and Methicillin-Resistant Staphylococcus aureus (MRSA) strains have also been reported with increasing frequency. MRSA strains have been associated with nosocomial as well as community acquired infections. Medicinal plants have enormous potential as antimicrobial substances and have been used in traditional medicine. Search for medicinally valuable plants with antimicrobial activity is being emphasized due to increasing antibiotic resistance in bacteria. In the present study, the antibacterial potential of Rosmarinus officinalis (Rosemary) and Murraya koenigii (curry leaves) was evaluated. These are common household herbs used in food as enhancer of flavor and aroma. The crude aqueous infusion, decoction and ethanolic extracts of curry leaves and rosemary and essential oil of rosemary were investigated in the present study for antibacterial activity against multi-drug resistant Staphylococcus strains using well diffusion method. In the present study, 60 Multi-drug resistant clinical isolates of S. aureus (43) and Coagulase Negative Staphylococci (CoNS) (17) were screened against different concentrations of crude extracts of Rosmarinus officinalis and Murraya koenigii. Out of these 60 isolates, 43 were sensitive to the aqueous infusion of rosemary; 23 to aqueous decoction and 58 to ethanolic extract whereas, 24 isolates were sensitive to the essential oil. In the case of the curry leaves, no antibacterial activity was observed in aqueous infusion and decoction while only 14 isolates were sensitive to the ethanolic extract. The aqueous infusion of rosemary (50% concentration) exhibited a zone of inhibition of 21(±5.69) mm. against CoNS and 17(±4.77) mm. against S. aureus, the zone of inhibition of 50% concentration of aqueous decoction of rosemary was also larger against CoNS 17(±5.78) mm. then S. aureus 13(±6.91) mm. and the 50% concentrated ethanolic extract showed almost similar zone of inhibition in S. aureus 22(±3.61) mm. and CoNS 21(±7.64) mm. whereas, the essential oil of rosemary showed greater zone of inhibition against S. aureus i.e., 16(±4.67) mm. while CoNS showed 15(±6.94) mm. These results show that ethanolic extract of rosemary has significant antibacterial activity. Aqueous infusion and decoction of curry leaves revealed no significant antibacterial potential against all Staphylococcal species and ethanolic extract also showed only a weak response. Staphylococcus strains were susceptible to crude extracts and essential oil of rosemary in a dose depend manner, where the aqueous infusion showed highest zone of inhibition and ethanolic extract also demonstrated antistaphylococcal activity. These results demonstrate that rosemary possesses antistaphylococcal activity.

Keywords: antibacterial activity, curry leaves, multidrug resistant, rosemary, S. aureus

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17 Screening for Larvicidal Activity of Aqueous and Ethanolic Extracts of Fourteen Selected Plants and Formulation of a Larvicide against Aedes aegypti (Linn.) and Aedes albopictus (Skuse) Larvae

Authors: Michael Russelle S. Alvarez, Noel S. Quiming, Francisco M. Heralde


This study aims to: a) obtain ethanolic (95% EtOH) and aqueous extracts of Selaginella elmeri, Christella dentata, Elatostema sinnatum, Curculigo capitulata, Euphorbia hirta, Murraya koenigii, Alpinia speciosa, Cymbopogon citratus, Eucalyptus globulus, Jatropha curcas, Psidium guajava, Gliricidia sepium, Ixora coccinea and Capsicum frutescens and screen them for larvicidal activities against Aedes aegypti (Linn.) and Aedes albopictus (Skuse) larvae; b) to fractionate the most active extract and determine the most active fraction; c) to determine the larvicidal properties of the most active extract and fraction against by computing their percentage mortality, LC50, and LC90 after 24 and 48 hours of exposure; and d) to determine the nature of the components of the active extracts and fractions using phytochemical screening. Ethanolic (95% EtOH) and aqueous extracts of the selected plants will be screened for potential larvicidal activity against Ae. aegypti and Ae. albopictus using standard procedures and 1% malathion and a Piper nigrum based ovicide-larvicide by the Department of Science and Technology as positive controls. The results were analyzed using One-Way ANOVA with Tukey’s and Dunnett’s test. The most active extract will be subjected to partial fractionation using normal-phase column chromatography, and the fractions subsequently screened to determine the most active fraction. The most active extract and fraction were subjected to dose-response assay and probit analysis to determine the LC50 and LC90 after 24 and 48 hours of exposure. The active extracts and fractions will be screened for phytochemical content. The ethanolic extracts of C. citratus, E. hirta, I. coccinea, G. sepium, M. koenigii, E globulus, J. curcas and C. frutescens exhibited significant larvicidal activity, with C. frutescens being the most active. After fractionation, the ethyl acetate fraction was found to be the most active. Phytochemical screening of the extracts revealed the presence of alkaloids, tannins, indoles and steroids. A formulation using talcum powder–300 mg fraction per 1 g talcum powder–was made and again tested for larvicidal activity. At 2 g/L, the formulation proved effective in killing all of the test larvae after 24 hours.

Keywords: larvicidal activity screening, partial purification, dose-response assay, capsicum frutescens

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16 Bioefficacy of Catharanthus roseus on Reproductive Performance of Red Cotton Bug, Dysdercus koenigii (Heteroptera: Pyrrhocoriedae)

Authors: Sunil Kayesth, Kamal Kumar Gupta


Influence of hexane extract of Catharanthus roseus leaves on reproductive fitness of Dysdercus koenigii was investigated by evaluating mating behaviour, oviposition behaviour and fertility of the treated insects. The volatiles of the plants were extracted in hexane by ‘cold extraction method’. The insects were treated with the extracts by ‘dry film residual method’. Our studies indicated that the treated male showed altered courtship behaviour, less number of mounting attempts, took more time to mate, less percent successful mating, and more disrupted mating. Similarly, the treated female exhibited either mating refusal or neutral behaviour towards courting males. The maximum disruption in the mating was observed in a cross T♂ X T♀, where males and females were treated with Catharanthus extract. The Dysdercus treated with Catharanthus extracts also showed marked reduction in their reproductive success. The treated females laid lesser number of egg batches and eggs in their life span. Catharanthus extract was effective in alteration of the oviposition behaviour. The eggs laid by the mated females were fertile indicating insemination of the mated females. However, the percent hatchability of the eggs laid by the treated females was less than control. The GC-MS analysis of the extract revealed the presence of juvenile hormone mimics, and the intermediates of juvenile hormone biosynthesis. Therefore, some of these compounds individually or synergistically alter reproductive behaviour of Dysdercus.

Keywords: Catharanthus roseus, Dysdercus koenigii, GC-MS analysis, reproductive performance

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15 Somatic Hybridization of between Citrus and Murraya paniculata Cells Applied by Electro-Fusion

Authors: Hasan Basri Jumin


Protoplasts isolated from embryogenic callus of Citrus sinensis were electrically used with mesophyll protoplasts isolated from seedless Citrus relatives. Hybrid of somatic embryos plantlets was obtained after 7 months of culture. Somatic hybrid plants were regenerated into normal seedlings and successfully transferred to soil after strictly acclimatization in the glass pot. The somatic hybrid plants were obtained by screening on the basis of chromosomes count. The number of chromosome of root tip counting revealed plantlets tetraploids (2n = 4x = 36) and the other were diploids (2n = 2x = 18) morphologically resembling the mesophyll parent. This somatic hybrid will be utilized as a possible pollen parent for improving the Citrus sinensis. A complete protoplast-to-plant system of somatic hybrid was developed for Citrus sinensis and Citrus relatives which could facilitate the transfer of nuclear and cytoplasmic genes from this species into cultivated Citrus through protoplast fusion.

Keywords: chromosome, Murraya paniculata, protoplast fusion, somatic hybrid, tetrapoliod

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14 Mating Behaviour and Its Significance in Reproductive Performance of Dysdercus koenigii

Authors: Kamal Kumar Gupta


The present research work was carried out on Dysdercus koenigii to understand various aspects of reproductive behavior such as mate finding and recognition, mate selection and mating preference, mating receptivity, and prolonged copulation. The studies carried out on mate searching and courtship behaviour of Dysdercus reflected the courtship behaviour in Dysdercus was brief. The opposite sexes are brought together by the pheromone. The males responded to female sex pheromones by showing directional movements toward the sex partners. Change in mating receptivity pattern of female Dysdercus was ascertained using three parameters of mating behaviour i.e. numbers of male’s encounter, the time taken to mate successfully and per cent females responding to mating. It was seen that a receptive female responded positively to the courting males and a high percentage of females mate usually in a very short time span. The females of Dysdercus showed continued mating receptivity throughout their life. The studies pertaining to mate selection by females showed that females generally do not discriminate among males and usually mate with any male they encountered first. The adults of Dysdercus remain in continuous copula up to 72hr. and mate 5-7 time in their life span. Studies pertaining to significance of prolonged mating in the life time reproductive success of the female Dysdercus indicated that fecundity and fertility and oviposition behavior of the female Dysdercus was related to duration of mating. In order to understand sperm precedence, the sterilized males were produced by exposing them to Gamma radiation. Our studies indicated that a dose of 50 Gy of Gamma radiations induced 95% sterility but does not impair the mating behaviour drastically. To understand role of sperms which were transfer during second mating in fertilizing the subsequent egg batches the sperm utilization pattern of doubly mated female was assessed. The females were mated with normal male or sterilized male in a combination. The sperm utilization pattern was determined by P2 value, our studies indicated a very high P2 value of 0.966, and indicated that sperms of last mating were utilized by the female for fertilization. In light of some of the unique reproductive behaviour of Dysdercus koenigii, such as brief courtship behavior, generalized mate selection by the female, continued mating receptivity and a prolonged pre oviposition period, the present studies on sperm precedence provides an explanation to an unusually prolonged copulation in Dysdercus.

Keywords: dysdercus koenigii, mating behaviour, reproductive performance, entomology

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13 Influence of Farnesol on Growth and Development of Dysdercus koenigii

Authors: Shailendra Kumar, Kamal Kumar Gupta


Dysdercus koenigii is an economically important pest of cotton worldwide. The pest damages the crop by sucking sap, staining lint, reducing the oil content of the seeds and deteriorating the quality of cotton. Plant possesses a plethora of secondary metabolites which are used as defense mechanism against herbivores. One of the important categories of such chemicals is insect growth regulators and the intermediates in their biosynthesis. Farnesol belongs to sesquiterpenoid. It is an intermediate in Juvenile hormone biosynthetic pathway in insects has been widely reported in the variety of plants. This chemical can disrupt the normal metabolic function and therefore, affects various life processes of the insects. Present study tested the efficacy of farnesol against Dysdercus koenigii. 2μl of 5% (100µg) and 10% (200µg) of the farnesol was applied topically on the dorsum of thoracic region of the newly emerged fifth instar nymphs of Dysdercus. The treated insects were observed daily for their survival, weight gain, and developmental anomalies for a period of ten days. The results indicated that treatment with 200µg farnesol decreased survival of the insects to 70% after 24h of exposure. At lower doses, no significant decrease in the survival was observed. However, the surviving nymphs showed alteration in growth, development, and metamorphosis. The weight gain in the treated nymphs showed deviation from control. The treated nymphs showed an increase in mortality during subsequent days and increase in the nymphal duration. The number of nymphs undergoing metamorphosis decreased to 46% and 88% in the treatments with the dose of 200µg and 100µg respectively. Severe developmental anomalies were also observed in the treated nymphs. The treated nymphs moulted into supernumerary nymphs, adultoids, adults with exuviae attached and adults with wing deformities. On treatment with 200µg; 26% adultoid, 4% adults with exuviae attached and 12% adults with wing deformed were produced. Treatment with 100µg resulted in production of 34% adultoid, 26% adults with deformed wing and 4% adults with exuviae attached. Many of the treated nymphs did not metamorphose into adults, remained in nymphal stage and died. Our results indicated potential application plant-derived secondary metabolites like farnesol in the management of Dysdercus population.

Keywords: development, Dysdercus koenigii, farnesol, survival

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12 Influence of Catharanthus roseus, Ocimum sanctum and Lantana camara Extracts on Survival and Longevity of Dysdercus koenigii

Authors: Sunil Kayesth, Kamal Kumar Gupta


The development of resistance among insects and pests, environmental contamination and adverse effects on non-target organisms is contributed by the indiscriminate use of chemical based insecticides. To overcome these environmental and other ecological issues that are need to replace these harmful toxic compounds. The present study was designed to evaluate the effect of Catharanthus roseus, Ocimum sanctum and Lantana camara plants volatiles on survival and longevity of Dysdercus koenigii. The hexane extract and ethanol extracts of these three plants were used. The fifth instars were exposed to hexane extract with concentrations of 10%, 5%, 2.5% 1.25%, 0.1%, 0.5% 0.25%, 0.125% and 0.0625% while, adults were treated with10%, 5%, 2.5% and 1.25%. 1-ml of each of these concentrations was used to make a thin film in sterilized glass jars of 500 ml capacity. Fifteen- newly emerged fifth instar nymphs and adult bugs were treated separately with the extracts for 24- hour exposure to the plant volatiles. For ethanol extracts cottonseed were treated with ethanol extracts of 10%, 5%, 2.5% and 1.25% concentrations. The treated seeds were provided to the Dysdercus for a period of 24 hours and their feeding behaviour was observed. The effect of hexane and ethanol extract of these plants was observed and readings were recorded for 15 days. Survival and longevity of both fifth instars and adults were in correlation with the concentrations of the plant extracts. Among three plant extracts, Ocimum hexane extract was most toxic and Catharanthus was moderate while Lantana was least toxic. The ethanol extracts of Lantana was highly antifeedent while Ocimum was moderate and Catharanthus was least antifeedent. Both Catharanthus and Ocimum appeared to have potential molecules, which possessed insecticidal activity while Ocimum and Lantana showed antifeedent activities. These insecticidal and antifeedent properties may be used in IPM.

Keywords: Catharanthus roseus, Ocimum sanctum, Lantana camara, Dysdercus koenigii

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11 Protoplast Cultures of Murraya paniculata L. Jack and Their Regeneration into Plant Precocious Flowering

Authors: Hasan Basri Jumin


Protoplasts isolated from embryogenic callus of Murraya paniculata (L. Jack.) were cultured in MT (Murashige and Tucker, 1969) basal medium containing 5% sucrose supplemented with kinetin, malt extract (ME) and 0.6 M sorbitol. About 85% of the surviving protoplasts formed a cell wall within 6 d of culture and the first cell division was observed 7 days after isolation. The highest plating effi¬ciency was obtained on MT basal medium containing 5% sucrose supplemented with 0.01 mg 1-1 kinetin 600 mg 1-1 ME, MT basal medium containing 5% sucrose and supplemented with 0.01 mg 1-1 Indole-acetic-acid (IAA) was found to be a medium suitable for the development somatic embryos into heart-shaped somatic embryos. The highest percentage of shoot formation was obtained using 0.1 mg 1-1 Indole-acitic-acid (IAA) 0..1 mg 1-1 gibberellic acid (GA3). In this investigation 40 plants were survived and grew normally in the soil. After two months maitained in the soil plants formed flower and flower developed into fruits on the soil treated with BA.

Keywords: gibberellic-acid, indole-acetic-acid, protoplast, precocious-flowering, somatic-embryo

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10 Somatic Embryogenesis Derived from Protoplast of Murraya Paniculata L. Jack and Their Regeneration into Plant Flowering in vitro

Authors: Hasan Basri Jumin


The in vitro flowering of orange jessamine plantlets derived from protoplast was affected by the manipulation of plant growth regulators, sugar and light conditions. MT basal medium containing 5% sucrose and supplemented with 0.001 mg 1-1 indole-acetic-acid was found to be a suitable medium for development of globular somatic embryos derived from protoplasts to form heart-shaped somatic embryos with cotyledon-like structures. The highest percentage (85 %) of flowering was achieved with plantlet on half-strength MT basal medium containing 5% sucrose and 0.001 mg1-1 indole-acetic-acid in light. Exposure to darkness for more than 3 weeks followed by re-exposure to light reduced flowering. Flowering required a 10-day exposure to indole-acetic-acid. Photoperiod with 18 h and 79.4 µmol m-2 s-1 light intensity promoted in vitro flowering in high frequencies. The sucrose treatment affected the flower bud size distribution. Flower buds originating from plantlet derived from protoplasts developed into normal flowers.

Keywords: indole-acetc-acid, light-intensity, Murraya-paniculata, photoperiod, plantlet, Zeatin

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9 Bioefficacy of Ocimum sanctum on Reproductive Performance of Red Cotton Bug, Dysdercus koenigii (Heteroptera: Pyrrhocoriedae)

Authors: Kamal Kumar Gupta, Sunil Kayesth


Dysdercus koenigii is serious pest of cotton and other malvaceous crop. Present research work aimed at ecofriendly approach for management of pest by plant extracts. The impact of Ocimum sanctum was studied on reproductive performance of Dysdercus koenigii. The hexane extract of Ocimum leaves was prepared by ‘cold extraction method’. The newly emerged fifth instar nymphs were exposed to the extract of concentrations ranging from 0.1% to 0.00625% by ‘thin film residual method’ for a period of 24h. Reproductive fitness of the adults emerged from the treated nymphs was evaluated by assessing their courtship behaviour, oviposition behaviour, and fertility. The studies indicated that treatment of Dysdercus with the hexane extract of Ocimum altered their courtship behaviour. Consequently, the treated males exhibited less sexual activity, performed fewer mounting attempts, increased time to mate and showed decreased percent successful mating. The females often rejected courting treated male by shaking the abdomen. Similarly, the treated females in many cases remained non-receptive to the courting male. Premature termination of mating in the mating pairs prior to insemination further decreased the mating success of the treated adults. Maximum abbreviation of courtship behaviour was observed in the experimental set up where both the males and the females were treated. Only females which mate successfully were observed for study of oviposition behaviour. The treated females laid lesser number of egg batches and eggs in their life span. The eggs laid by these females were fertile indicating insemination of the female. However, percent hatchability was lesser than control. The effects of hexane extract were dose dependent. Treatment with 0.1% and 0.05% extract altered courtship behaviour. Doses of concentrations less than 0.05% did not affect courtship behaviour but altered the oviposition behaviour and fertility. Significant reduction in the fecundity and fertility was observed in the treatments at concentration as low as 0.00625%. The GCMS analysis of the extract revealed a plethora of phytochemicals including juvenile hormone mimics, and the intermediates of juvenile hormone biosynthesis. Therefore, some of these compounds individually or synergistically impair reproductive behaviour of Dysdercus. Alteration of courtship behaviour and suppression of fecundity and fertility with the help of plant extracts has wide potentials in suppression of pest population and ‘integrated pest management’.

Keywords: courtship behaviour, Dysdercus koenigii, Ocimum sanctum, oviposition behaviour

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8 Quest for an Efficient Green Multifunctional Agent for the Synthesis of Metal Nanoparticles with Highly Specified Structural Properties

Authors: Niharul Alam


The development of energy efficient, economic and eco-friendly synthetic protocols for metal nanoparticles (NPs) with tailor-made structural properties and biocompatibility is a highly cherished goal for researchers working in the field of nanoscience and nanotechnology. In this context, green chemistry is highly relevant and the 12 principles of Green Chemistry can be explored to develop such synthetic protocols which are practically implementable. One of the most promising green chemical synthetic methods which can serve the purpose is biogenic synthetic protocol, which utilizes non-toxic multifunctional reactants derived from natural, biological sources ranging from unicellular organisms to higher plants that are often characterized as “medicinal plants”. Over the past few years, a plethora of medicinal plants have been explored as the source of this kind of multifunctional green chemical agents. In this presentation, we focus on the syntheses of stable monometallic Au and Ag NPs and also bimetallic Au/Ag alloy NPs with highly efficient catalytic property using aqueous extract of leaves of Indian Curry leaf plat (Murraya koenigii Spreng.; Fam. Rutaceae) as green multifunctional agents which is extensively used in Indian traditional medicine and cuisine. We have also studied the interaction between the synthesized metal NPs and surface-adsorbed fluorescent moieties, quercetin and quercetin glycoside which are its chemical constituents. This helped us to understand the surface property of the metal NPs synthesized by this plant based biogenic route and to predict a plausible mechanistic pathway which may help in fine-tuning green chemical methods for the controlled synthesis of various metal NPs in future. We observed that simple experimental parameters e.g. pH and temperature of the reaction medium, concentration of multifunctional agent and precursor metal ions play important role in the biogenic synthesis of Au NPs with finely tuned structures.

Keywords: green multifunctional agent, metal nanoparticles, biogenic synthesis

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7 Ecofriendly Approach for the Management of Red Cotton Bug Dysdercus koenigii by Botanicals

Authors: S: Kayesth, K. K. Gupta


The indiscriminate use of insecticides causes environmental contamination, adversely affects non-target organisms and develops resistance among insects and pests. There has always been felt a need for methods of control which can overcome these environmental and other ecological issues. The present study was designed to evaluate the effect of different plants volatiles on survival, longevity, growth, development and reproduction of Dysdercus koenigii. The hexane extract of three different plants (Catharanthus roseus, Ocimum sanctum and Lantana camara) was used. The fifth instars were exposed to hexane extract with concentrations of 10%, 5%, 2.5%, 1.25%, 0.1%, 0.5%, 0.25%, 0.13% and 0.06% while adults were treated with 10%, 5%, 2.5% and 1.25%. 1-ml of each of these concentrations was used to make a thin film in sterilized glass jars of 500 ml capacity. Fifteen newly emerged fifth instar nymphs and ten pairs of adult bugs were treated separately with the extracts for 24 hour exposure to the plant volatiles. The effect of these plant extract was observed and readings were recorded for 23 days. Survival and longevity of both fifth instars and adults were in correlation with the concentrations of the plant extracts. The extracts did not influence growth of fifth instars significantly but impaired their development significantly at higher concentrations. The treated nymphs at higher concentrations either could not moult or died and those which could moult moulted into supranumery instars, adultoids or adults with wing deformities. The supranumery insects retained the nymphal characters except increased body size and wing pads. The adultoids had wing deformities and non-functional reproductive organs. Adultoids exhibited courtship and mounting attempts but were not able to mate. At lower concentrations from 0.1 to 0.06% the fifth instars developed into adults with fewer deformities. At these concentrations, the fecundity and fertility of these adults were drastically reduced. On the contrary, the treated adults also had reduced fecundity and fertility compared to control. Among three plant extracts Ocimcum was most toxic for both fifth instars and adults in terms of survival and longevity. Catharanthus, Ocimum and Lantana appeared to have potential molecules which possessed insect juvenile hormone like activity. Potential application of these plant extracts in IPM was discussed.

Keywords: Catharanthus, Ocimum, Lantana, Dysdercus koenigii

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6 Comparative Study of Antimicrobial, Antioxidant and Physicochemical Properties of Four Culinary Herbs Grown in Sri Lanka

Authors: Thilini Kananke


Culinary herbs have long been considered as significant dietary sources of many potential health-promoting compounds. The present research focused on analysis of antimicrobial, antioxidant and physicochemical properties in selected four culinary herbs namely Murraya koenigii (Curry leaves), Pandanus amaryllifolius (Pandan leaves), Cymbopogon citrates (Lemon grass leaves), and Mentha Piperita (Minchi leaves) obtained from several market sites in Ratnapura District, Sri Lanka. The antimicrobial activity of ethanolic, chloroform and distilled water extracts of culinary herbs were evaluated against the strains of Staphylococcus aureus, Salmonella typhi and Shigella spp. Total phenolic content and the radical scavenging activity (using DPPH assay) of culinary herbs were determined. Four heavy metals (Cu, Cd, Pb and Fe) were analyzed in the selected culinary herbs using the atomic absorption spectroscopy (AAS). Proximate compositions of the selected herbs were analyzed using AOAC official methods. Antimicrobial activity of all selected culinary herbs showed relativity high inhibition zones against S. aureus. Pandan leaves showed the least antimicrobial activity against selected bacterial strains compared with other culinary herbs. Both the highest radical scavenging activity (lower IC50 value) and the total phenolic content (25.57 ±3.54µg GAE/100g) were reported in Mentha piperita extract. The highest concentrations of Cu, Fe and Cd were reported in Curry leaves (29.15 mg/kg), Lemon grass leaves (257.98 mg/kg) and Pandan leaves (6.05 mg/kg) respectively. The heavy metal contents detected in all culinary herbs were below the permitted limits set by WHO/FAO, except Cd. The highest moisture (85.00±0.00%) and fiber (10.66± 2.00%) contents were found in Pandan leaves, while the highest protein (8.94±0.29%), fat (12.3± 2.52%) and ash (3.50± 0.17%) contents were reported in curry leaves. The information obtained from this study highlights the importance of further investigation of other antioxidant, antimicrobial and health promoting compounds of culinary herbs available in Sri Lanka for a detailed comparison.

Keywords: antimicrobial, antioxidant, culinary herbs, proximate analysis

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5 Acute Antihyperglycemic Activity of a Selected Medicinal Plant Extract Mixture in Streptozotocin Induced Diabetic Rats

Authors: D. S. N. K. Liyanagamage, V. Karunaratne, A. P. Attanayake, S. Jayasinghe


Diabetes mellitus is an ever increasing global health problem which causes disability and untimely death. Current treatments using synthetic drugs have caused numerous adverse effects as well as complications, leading research efforts in search of safe and effective alternative treatments for diabetes mellitus. Even though there are traditional Ayurvedic remedies which are effective, due to a lack of scientific exploration, they have not been proven to be beneficial for common use. Hence the aim of this study is to evaluate the traditional remedy made of mixture of plant components, namely leaves of Murraya koenigii L. Spreng (Rutaceae), cloves of Allium sativum L. (Amaryllidaceae), fruits of Garcinia queasita Pierre (Clusiaceae) and seeds of Piper nigrum L. (Piperaceae) used for the treatment of diabetes. We report herein the preliminary results for the in vivo study of the anti-hyperglycaemic activity of the extracts of the above plant mixture in Wistar rats. A mixture made out of equal weights (100 g) of the above mentioned medicinal plant parts were extracted into cold water, hot water (3 h reflux) and water: acetone mixture (1:1) separately. Male wistar rats were divided into six groups that received different treatments. Diabetes mellitus was induced by intraperitoneal administration of streptozotocin at a dose of 70 mg/ kg in male Wistar rats in group two, three, four, five and six. Group one (N=6) served as the healthy untreated and group two (N=6) served as diabetic untreated control and both groups received distilled water. Cold water, hot water, and water: acetone plant extracts were orally administered in diabetic rats in groups three, four and five, respectively at different doses of 0.5 g/kg (n=6), 1.0 g/kg(n=6) and 1.5 g/kg(n=6) for each group. Glibenclamide (0.5 mg/kg) was administered to diabetic rats in group six (N=6) served as the positive control. The acute anti-hyperglycemic effect was evaluated over a four hour period using the total area under the curve (TAUC) method. The results of the test group of rats were compared with the diabetic untreated control. The TAUC of healthy and diabetic rats were 23.16 ±2.5 mmol/L.h and 58.31±3.0 mmol/L.h, respectively. A significant dose dependent improvement in acute anti-hyperglycaemic activity was observed in water: acetone extract (25%), hot water extract ( 20 %), and cold water extract (15 %) compared to the diabetic untreated control rats in terms of glucose tolerance (P < 0.05). Therefore, the results suggest that the plant mixture has a potent antihyperglycemic effect and thus validating their used in Ayurvedic medicine for the management of diabetes mellitus. Future studies will be focused on the determination of the long term in vivo anti-diabetic mechanisms and isolation of bioactive compounds responsible for the anti-diabetic activity.

Keywords: acute antihyperglycemic activity, herbal mixture, oral glucose tolerance test, Sri Lankan medicinal plant extracts

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4 Farmers Perception in Pesticide Usage in Curry Leaf (Murraya koeinigii (L.))

Authors: Swarupa Shashi Senivarapu Vemuri


Curry leaf (Murraya koeinigii (L.)) exported from India had insecticide residues above maximum residue limits, which are hazardous to consumer health and caused rejection of the commodity at the point of entry in Europe and middle east resulting in a check on export of curry leaf. Hence to study current pesticide usage patterns in major curry leaf growing areas, a survey on pesticide use pattern was carried out in curry leaf growing areas in Guntur districts of Andhra Pradesh during 2014-15, by interviewing farmers growing curry leaf utilizing the questionnaire to assess their knowledge and practices on crop cultivation, general awareness on pesticide recommendations and use. Education levels of farmers are less, where 13.96 per cent were only high school educated, and 13.96% were illiterates. 18.60% farmers were found cultivating curry leaf crop in less than 1 acre of land, 32.56% in 2-5 acres, 20.93% in 5-10 acres and 27.91% of the farmers in more than 10 acres of land. Majority of the curry leaf farmers (93.03%) used pesticide mixtures rather than applying single pesticide at a time, basically to save time, labour, money and to combat two or more pests with single spray. About 53.48% of farmers applied pesticides at 2 days interval followed by 34.89% of the farmers at 4 days interval, and about 11.63% of the farmers sprayed at weekly intervals. Only 27.91% of farmers thought that the quantity of pesticides used at their farm is adequate, 90.69% of farmers had perception that pesticides are helpful in getting good returns. 83.72% of farmers felt that crop change is the only way to control sucking pests which damages whole crop. About 4.65% of the curry leaf farmers opined that integrated pest management practices are alternative to pesticides and only 11.63% of farmers felt natural control as an alternative to pesticides. About 65.12% of farmers had perception that high pesticide dose will give higher yields. However, in general, Curry leaf farmers preferred to contact pesticide dealers (100%) and were not interested in contacting either agricultural officer or a scientist. Farmers were aware of endosulfan ban 93.04%), in contrast, only 65.12, per cent of farmers knew about the ban of monocrotophos on vegetables. Very few farmers knew about pesticide residues and decontamination by washing. Extension educational interventions are necessary to produce fresh curry leaf free from pesticide residues.

Keywords: Curry leaf, decontamination, endosulfan, leaf roller, psyllids, tetranychid mite

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3 Evaluation of Physical Parameters and in-Vitro and in-Vivo Antidiabetic Activity of a Selected Combined Medicinal Plant Extracts Mixture

Authors: S. N. T. I. Sampath, J. M. S. Jayasinghe, A. P. Attanayake, V. Karunaratne


Diabetes mellitus is one of the major public health posers throughout the world today that incidence and associated with increasing mortality. Insufficient regulation of the blood glucose level might be serious effects for health and its necessity to identify new therapeutics that have ability to reduce hyperglycaemic condition in the human body. Even though synthetic antidiabetic drugs are more effective to control diabetes mellitus, there are considerable side effects have been reported. Thus, there is an increasing demand for searching new natural products having high antidiabetic activity with lesser side effects. The purposes of the present study were to evaluate different physical parameters and in-vitro and in-vivo antidiabetic potential of the selected combined medicinal plant extracts mixture composed of leaves of Murraya koenigii, cloves of Allium sativum, fruits of Garcinia queasita and seeds of Piper nigrum. The selected plants parts were mixed and ground together and extracted sequentially into the hexane, ethyl acetate and methanol. Solvents were evaporated and they were further dried by freeze-drying to obtain a fine powder of each extract. Various physical parameters such as moisture, total ash, acid insoluble ash and water soluble ash were evaluated using standard test procedures. In-vitro antidiabetic activity of combined plant extracts mixture was screened using enzyme assays such as α-amylase inhibition assay and α-glucosidase inhibition assay. The acute anti-hyperglycaemic activity was performed using oral glucose tolerance test for the streptozotocin induced diabetic Wistar rats to find out in-vivo antidiabetic activity of combined plant extracts mixture and it was assessed through total oral glucose tolerance curve (TAUC) values. The percentage of moisture content, total ash content, acid insoluble ash content and water soluble ash content were ranged of 7.6-17.8, 8.1-11.78, 0.019-0.134 and 6.2-9.2 respectively for the plant extracts and those values were less than standard values except the methanol extract. The hexane and ethyl acetate extracts exhibited highest α-amylase (IC50 = 25.7 ±0.6; 27.1 ±1.2 ppm) and α-glucosidase (IC50 = 22.4 ±0.1; 33.7 ±0.2 ppm) inhibitory activities than methanol extract (IC50 = 360.2 ±0.6; 179.6 ±0.9 ppm) when compared with the acarbose positive control (IC50 = 5.7 ±0.4; 17.1 ±0.6 ppm). The TAUC values for hexane, ethyl acetate, and methanol extracts and glibenclamide (positive control) treated rats were 8.01 ±0.66; 8.05 ±1.07; 8.40±0.50; 5.87 ±0.93 mmol/L.h respectively, whereas in diabetic control rats the TAUC value was 13.22 ±1.07 mmol/L.h. Administration of plant extracts treated rats significantly suppressed (p<0.05) the rise in plasma blood glucose levels compared to control rats but less significant than glibenclamide. The obtained results from in-vivo and in-vitro antidiabetic study showed that the hexane and ethyl acetate extracts of selected combined plant mixture might be considered as a potential source to isolate natural antidiabetic agents and physical parameters of hexane and ethyl acetate extracts will helpful to develop antidiabetic drug with further standardize properties.

Keywords: diabetes mellitus, in-vitro antidiabetic assays, medicinal plants, standardization

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2 A Comparative Study of Murayya Koenigii Varieties for the Removal of Cr (VI) from Aqueous Solutions

Authors: Mesfin Tsegaw, Sivakumar C. V., Chandrakal Gunturu, Meera Indracanti


Chromium (VI), a toxic metal ion, is widely used in electroplating, stainless steel production, leather tanning, paint, and textile manufacturing. Cr (VI) is mobile in the environment, acutely toxic and carcinogenic. In the present study, the ability to remove Cr (VI) from aqueous solutions has been compared using leaves of dwarf and gamthi varieties of Murayya koerigii abundantly available in Selaqui region of Dehradun as an adsorbent. Effects of temperature, pH, initial concentration of adsorbate and adsorbent dosage have been studied for effective removal of Cr (VI). The biosorptive ability of biosorbent was reliant on the pH of the biosorbate, with pH 2 being most favorable for both the varieties. The obtained results were analyzed by the Freundlich and Langmuir equation at different temperature and related parameters were determined for each adsorption isotherm. The study also includes results on the kinetic dimensions of adsorption of the Cr (VI) on the derived adsorbent. Gamthi variety has a promising absorption rate of 80% over the dwarf variety. FTIR studies confirmed that carboxyl and hydroxyl groups were the main groups involved in the metal uptake.

Keywords: adsorption, cromium, kinetics, variety

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1 Effect of Abiotic Factors on Population of Red Cotton Bug Dysdercus Koenigii F. (Heteroptera: Pyrrhocoridae) and Its Impact on Cotton Boll Disease

Authors: Haider Karar, Saghir Ahmad, Amjad Ali, Ibrar Ul Haq


The experiment was conducted at Cotton Research Station, Multan to study the impact of weather factors and red cotton bug (RCB) on cotton boll disease yielded yellowish lint during 2012. The population on RCB along with abiotic factors was recorded during three consecutive years i.e. 2012, 2013, and 2014. Along with population of RCB and abiotic factors, the number of unopened/opened cotton bolls (UOB), percent yellowish lint (YL) and whitish lint (WL) were also recorded. The data revealed that the population per plant of RCB remain 0.50 and 0.34 during years 2012, 2013 but increased during 2014 i.e. 3.21 per plant. The number of UOB were more i.e. 13.43% in 2012 with YL 76.30 and WL 23.70% when average maximum temperature 34.73◦C, minimum temperature 22.83◦C, RH 77.43% and 11.08 mm rainfall. Similarly in 2013 the number of UOB were less i.e. 0.34 per plant with YL 1.48 and WL 99.53 per plant when average maximum temperature 34.60◦C, minimum temperature 23.37◦C, RH 73.01% and 9.95 mm rainfall. During 2014 RCB population per plant was 3.22 with no UOB and YL was 0.00% and WL was 100% when average maximum temperature 23.70◦C, minimum temperature 23.18◦C, RH 71.67% and 4.55 mm rainfall. So it is concluded that the cotton bolls disease was more during 2012 due to more rainfall and more percent RH. The RCB may be the carrier of boll rot disease pathogen during more rainfall.

Keywords: red cotton bug, cotton, weather factors, years

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