marine algae for the cotton pest and disease management

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Recent trends in Agriculture, Water and Environment Research Recent trends in Agriculture, Water and Environment Research Recent trends in Agriculture, Water and Environment Research Recent trends in Agriculture, Water and Environment Research

The SciTech Publishers & International Association for Teaching and Learning, 2012

MARINE ALGAE FOR THE COTTON PEST AND DISEASE MANAGEMENT

K. SAHAYARAJ 1, S. RAJESH 1, A. ASHA 1, AND J.M. RATHI 2 1 CROP PROTECTION RESEARCH CENTRE, DEPARTMENT OF ADVANCED ZOOLOGY

AND BIOTECHNOLOGY, ST. XAVIERS COLLEGE (AUTONOMOUS), PALAYAMKOTTAI

627 002, TAMIL NADU, INDIA,2 DEPARTMENT OF CHEMISTRY, ST. MARYS COLLEGE, THOOTHUKUDI 628 001,

TAMIL NADU, INDIA

Abstract:

Macroscopic marine algae, popularly known as seaweeds, form one of the importantliving resources of the ocean. Agar, carrageenan and alginate are popular examples ofseaweedsthese have been used as food for human beings, feed for animals, fertilizers for

plants and source of various chemicals. In the recent past, seaweeds have been gainingmomentum as new experimental systems for biological research and integrated aquaculture

systems. Totally 57 taxa belonging to 37 genera representing Chlorophyceae (17 species),Phaeophyceae (14 species) and Rhodophyceae (25 species) were recorded from the 19sampling sites during our study period (June 2009 to June 2010). More number of algae wererecorded from the Bay of Bengal (67.7%) followed by Indian Ocean (25%) and Arabian Seacoasts (8%).Among them, Caulerpa scalpelliformis (CS), Caulerpa veravalensis (CV), Ulvafasciata (UF), Ulva lactuca (UL), (Chlorophyta) Padina pavonica (PP), and Sargassumwightii (SW) were tested against Dysdercus cingulatus (Fab.) and fungal pathogen, Fusariumoxysporum f. sp. vasinfectum (Atk.) Snyd & Hans. at different concentrations. Extractionswere carried out using hot continuous extraction and cold Percolation methods using polar(water-AQ and methanol-ME) and non-polar solvents (Chloroform-CL and Hexane-HE).

Steroids, tannins, flavonoids were observed in AQ extract of CS, PP, ST and CC. However,alkaloids, pholobatanins (except in AQ extract of ST), aromatic acids were not recorded inthese algae. Soxhlation method can be used for the extraction of steroids, tannins, saponins,cardiac glycosides and phenolic compounds. Total tannins, Phenolic compounds, BoundPhenol and O.D. Phenol content was found higher in Sargassum wightii, Padinatetrastomatica, and Chaetomorpha crassa, respectively.It is concluded that tested algalseaweeds possess nymphicidal and adultoid, ovicidal, ovipositional, bactericidal andfungicidal activities. CV, CS and UF have nymphicidal and ovicidal activities, whereas CVhas adultoid activity and both CV and CS ovipositional activity. Hence Caulerpa veravalensischloroform extracts could be used for the red cotton bug management in cotton. CVmethanol, Chloroform and Hexane have similar bactericidal activity, whereas CV chloroformand CS methanol extract has fungicidal activity. The study suggested that CV can be utilizedfor management of both bacterial and fungal diseases of cotton. All these algae synthesizedsilver-based green nanoparticles. These green nanoparticles reduce the radish seedgermination rate by 36%. However, they did not affect the cotton, cucumber and tomato seedgermination and could be suggested to utilize these weeds in the agriculture.


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Keywords : Macroscopic marine algae, extraction, Dysdercus cingulatus, Fusariumoxysporum f. sp. vasinfectum, nanoparticles, agriculture use

Introduction:In India, cotton production is about 295.0 million (= 480-pound bales) during 2009-

2010, as against 113.9 million bales in the world. India also has the largest area under cottoncultivation (10.31 million hectares), and yield is 486 kg/ha during 2009-2010(http://cotcorp.gov.in). Cotton is damaged by over 160 species of insects from the seedingstage right up to the entire period of the plant growth.

The cotton stainer, Dysdercus cingulatus (Fab.) (Heteroptera: Pyrrhocoridae), causesserious damage by feeding on developing cotton bolls and ripe cotton seeds and transmittingfungi that develop on the immature lint and seeds (Natarajan and Rajendran, 2005).Previously, Rajendran and Gopalan (1980) studied the impact of Catharanthus roseus (L.) G.Don. (Astraceae), Parthenium hysterophorus L. (Apocynaceae) and Nephrolepis exaltata (L.)Schott (Nephrolepidaceae) extracts on morphological changes of D. cingulatus . The impactof different neem parts extracts on mortality of D. cingulatus has also been studied (Sharma

et al., 2010). Pedalium murex (L.) (Sahayaraj et al ., 2006) and Streblus asper Lour. (Hashimand Devi, 2003) root extracts prolong mating duration and reduced fecundity, hatchability,adult longevity of D. cingulatus . Moreover, more than 2500 terrestrial plants have beenscreened against agricultural pests; however, scientists developed insecticides only from theneem.

Wilt of cotton ( Gossypium spp) is a important vascular disease caused by the soil borne pathogen Fusarium oxysporum Schlechtend f.sp. vasinfectum (Atk.) Snyd and Hans.The disease is widespread and causes substantial crop losses in most of the major cotton-

producing areas of the world (Assigbeste et al., 1994; Wang et al., 2004). Due to hazardsassociated with the increased use of synthetic pesticides the use of biopesticides esp., from

marine algae has gained considerable attention on the eco-friendly approaches for themanagement of insect pest and plant pathogens.

Apart from terrestrial plants now days seaweeds have been used for the pestmanagement program. Dureja (1993), Ara et al. (1997), Rizvi (2003), Rizvi and Shameel(2003, 2004) highlighted the importance of algal seaweeds in insect pest management. Biju etal. (2004), Manilal et al. (2009), and Sahayaraj and Kalidas (2011) have recorded theinsecticidal activity of seaweeds like Bryopsis plumosa (Huds), Padina pavonica (Linn) and

Hyblaea puera (Cramer) on D. cingulatus and Culex quinquefasciatus respectively. Weselected Caulerpa veravalensis, Caulerpa scalpelliformis, Padina pavonica, Sargas sumwightii, Ulva fasciata and Ulva lactuca for this study. All these plants were available in

plenty and moreover, drifted seaweeds are merely a waste in many parts of the world and itcan be utilized for pest management program. Furthermore, a critical literature survey revealsthat all these plants have less explore or not been studied for its pesticidal property on anyagriculture pests. Hence, it is imperative to evaluate the insecticidal activity of marine plants.The objective of our present study was aimed to explore the impacts of C. veravalensis, C.

scalpelliformis, P. pavonica, S. wightii, U. fasciata and U. lactuca extracts against thedevastating, notorious cotton pest, D. cingulatus under laboratory conditions.


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Materials and methods:Collection and extraction of seaweeds:

Drifted C. veravalensis, C. scalpelliformis, P. pavonica, S. wightii, U. fasciata and U.lactuca were collected from coastal area of Kanyakumari, Thoothukudi, Tirunelveli Districts,Tamil Nadu, India. Immediately after collection, alga was washed in sea water; epiphytes,associated organisms, sands and other extraneous matter were removed. After subsequentwashing in fresh water, the plants shade dried for two weeks continuously. Then partially

powdered using domestic blender and stored in air tight container until when needed. Fromthe stock, 500 g of powdered material was extracted successively using benzene, (BN),chloroform (CH), hexane and water using Soxhlet apparatus continuously for 24 h at 50 C.The extract was concentrated with distillation apparatus at 40 C and again concentratedusing vacuum desiccators at room temperature to obtain minimum quantity of crude extractfor testing insecticidal activity on D. cingulatus. The extracts have also been tested against

Fusarium oxysporum f. sp. vasinfectum (Atk.) Snyd & Hans.

Pest collection and maintenance: Dysdercus cingulatus nymphs and adults were collected from cotton fields,

Tirunelveli district, Tamil Nadu, India, and subsequently maintained in the laboratory at 28 2 C and 70-75% RH on water soaked cotton seeds and fresh cotton leaves. The nymphsemerged from the laboratory laid egg masses were reared using cotton plants and newlyemerged third instar nymphs were used for the experiments. Each treatment contains sixreplicates and 10 insects were used for each replicate. Seed dip method of Sahayaraj andKalidas (2011) was followed for the insecticidal activity bioassay.

Insecticidal bioassay:Five concentrations (0.1%, 0.2%, 0.4%, 0.8%, and 1.6%) were prepared using 1 mL of

respective solvents, then diluted with 10 mL water and used for the study. Cotton seeds (100g) were separately taken in a conical flask and add 250 mL of plant extract and 3 mL ofTween 80 (0.1%) as an adjuvant. The flask was agitated at 65 rpm in a shaker (Remi,Mumbai) for 12 h at room temperature and provided as food to D. cingulatus . Ten third instar

D. cingulatus nymphs were taken in a plastic container (300 mL capacity) which coveredwith aerated lid. Control category was provided with water mixed with adjuvant soakedcotton seeds. Both for experiment and control categories, cotton seeds were replaced everyday by a new plant extracts soaked seeds for 4 d continuously. Mortality was recorded at 24,48, 72, and 96 h.

Antimicrobial activity: Fusarium oxysporum f.sp. vasinfectum was isolated from infected cotton plants

(Melameignanapuram, Tenkasi district, Tamil Nadu, India) and were used for the experiment.The pathogen was isolated, sub-cultured on Potato Dextrose Agar (PDA) medium andidentified using standard protocol (Burgess et al ., 1994). Antifungal activity was carried outusing agar well diffusion method (Irobi et al ., 1996). Petri plates were prepared with 20 ml ofsterile PDA. Wells were made using sterile cork borer under aseptic condition. The C.

scalpelliformis extract with various concentrations (0.05%, 0.1%, 0.2%, 0.4% and 0.8%)were prepared using Dimethyl Sulphoxide (DMSO) and were added to the respective wells.Carbendazim (Bavistin) (0.03%) was used as positive control and DMSO was maintained asnegative control. They were incubated at 27C for 3 days. The zone of inhibition wasmeasured using a ruler and expressed in mm.


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Secondary metabolites analysis :Both qualitative and quantitative [total phenols ( g mg -1), Ortho dihydric phenols (ODP)

phenol and bound phenols (BP) (mg g -1), and tannins ( g mg -1) phytochemical analysis of theextracts was carried out following the method of Harbone (1998).

Synthesis and characterization of liquid nanoparticles:Exactly 17 mg of AgNO 3 was dissolved in 100 mL distilled water (10

-3M). Ten mL ofalgal thallus extract was added to 90 mL of 10 -3M AgNo 3 solution for reduction of Ag

+ ions.The reduction of pure Ag + ions was monitored by measuring the UV-vis spectra of thesolution at regular intervals after diluting a small aliquot (0.2 mL) of the sample 20 times.UV-vis spectra were recorded as a function of time of reaction on a UV- 1601Shimadzuspectrophotometer with samples in Quartz cuvette operated at a resolution of 1 nm. Theliquid nanoparticle was kept as such for two months at room temperature (30-32C), then X ray diffraction (XRD) pattern of the alga thallus broth reduced Ag nanoparticles wereobtained using Siemens D5005 XRD (X- ray diffractometer) with Cu K radiation ( =0.1542). XRD patterns were analyzed to determine peak intensity, position and width. The

particle size was calculated using the Scherrer formula,d = 0.9 / cos

where, d is the mean diameter of the nanoparticles, , the wavelength of X-ray radiationsource and , the angular FWHM of the XRD peak at the diffraction angle (Culity, 1978).

The alga thallus broth reduced Ag nanoparticles solution was centrifuged at 13,000rpm for 15 minutes, redispersed in sterile distilled water to get rid of any uncoordinated

biological molecules for Fourier transform infrared (FTIR) spectroscopy measurements.Centrifugation and the redispersion were repeated thrice in order to ensure better separation.The purified KBr pellets were then air dried at room temperature and powdered subjected toFTIR spectroscopy measurement (Shimadzu FTIR-8300S). The morphology of the algathallus broth reduced Ag nanoparticles was recorded using the JSM-6390 Scanning electronmicroscope (SEM). Samples for SEM were prepared by drop coating the Ag nanoparticlessolutions onto carbon copper grid. The films on the grids were allowed to dry prior to SEMmeasurement. To record the size and shape of alga thallus broth reduced Ag nanoparticle,samples for Transmission Electron Microscopy (TEM) were prepared by drop-coating the Agnanoparticle solution onto carbon-coated copper grids. The films on the TEM grids wereallowed to stand for two minutes, following which the extra solution was removed using a

blotting paper and the grid allow drying prior to measurement. TEM measurements were performed on a JEOL model 3010 instrument operated at an accelerating voltage at 120 kv.

Statistical analysis:All results were expressed in mean with standard errors. Individual data was subjected

to one-way ANOVA and post ANOVA Tukey Multiple Range Test (TMRT); thesignificances are expressed at 5% level.


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Results and discussion :Macroscopic marine algae form one of the important living resources of the ocean

throughout the world and particularly in India (Anonymous, 1978, 1985, 2000, 2005; SubbaRao and Mantri, 2006). Agar, carrageenan and alginate are popular examples of seaweeds these have been used as food for human beings, feed for animals, anti-insect, anti-microbials,

anti-helmonthics, fertilizers for plants and source of various chemicals (Mshigeni, 1992;Anggadiredja, 1993; Afaq-Husain et al., 2001; Freile-Pelargin and Morales, 2004;Dhargalkar and Neelam Pereira, 2005; Sabina et al. (2005); Salvador et al., 2007; Ayson etal., 2008).

Distribution:One thousand and fifty specimens of algal seaweeds were collected by our team. The

present annotated checklist of 57 taxa belonging to 37 genera representing Chlorophyceae (17species), Phaeophyceae (14 species) and Rhodophyceae (25 species) (Table 1) were recordedfrom the study areas (Figure 2). The red algae dominated over green and brown algae.Presence of the rocky coasts of Tamil Nadu is abounding with a rich algal growth.Desikachary et al. (1990, 1998) also reported that Rhodophyceae are the common marinealgae in Tamil Nadu, India as observed here.

Caulerpa scalpelliformis was recorded from Circular Fort (N 0807'37.9', E07734'02.7'), Kootapuli (N 0808'44.2', E 07736'02.5'), Idinthakarai, Kuthenkuzhi,Tuticorin, Therkukalmaedu, Tiruchendur and Mandapam. This species was found throughoutthe year with high population during March to June, 2010. Similar trend was also observedfor Caulerpa veravalensis, Chaetomorpha crassa and Sargassum wightii . Sargassum wightii was found abundant in Kanyakumari, C. veravalensis in Idinthakarai and Kuthenkuzhi,Chaetomorpha crassa in Circular Fort, they were found attached to the rock. Chaetomorphacrassa was found to grow over Sargassum sp. Padina pavonica found abundant from June toAugust 2009 in Tuticorin and Mandapam. Padina tetrastromatica was abundant in CircularFort during March 2010. Lobophora variegata was recorded abundant in Circular Fort onlyduring April 2010. Similarly, Amphiroa anceps was abundantly collected only in Kootapuliduring April 2010. Ulva lactuca was found abundant in Tuticorin (June to August 2009),Ulva fasciata in Muttam, Idinthakarai and Kuthenkuzhi (September to November 2009).

Insecticidal activity:The toxicity of algal extracts was evaluated against D. cingulatus third instar nymphs

to suggest a safe method for their control. The percentage of mortality increased when theconcentration level increased. Chloroform extracts of Caulerpa veravalensis, Caulerpa

scalpelliformis, Padina pavonica, and Sargassum wightii and methanol extracts of Ulva fasciata and Ulva lactuca were found to have maximum nymphicidal activity against thirdinstar nymphs of Dysdercus cingulatus. Based up on the LC 50 values, it was concluded thatamong the six algal seaweeds, Caulerpa veravalensis chloroform extracts considered as the

best insecticidal algae followed by Caulerpa scalpelliformis, Ulva fasciata (methanol) , Ulvalactuca (methanol) , Sargas sum wightii (chloroform) and Padina pavonica (chloroform)(Table 2) .


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The algal extracts mixed with artificial diet might enter into the alimentary canalwhile feeding, affects the digestive physiology which lead to the death of the insect. Cetin etal. (2010) reported the larvicidal efficacy of the acetone extract of the thalli of Caulerpa

scalpelliformis var. denticulata against late second to early third instars of Culex pipiens at 1,200 ppm, the extract caused >70% larval mortality at 24-h, 48-h, and 72-h exposure. The

LC 50 (median lethal concentration) and LC 90 values of C. scalpelliformis were 338.91 and1,891.31 ppm, respectively. Bai and Koshy et al. (2004) reported that 40% leaf and 10% seedethanolic extracts of Thevetia neriifolia Juvenomimetic activity on D. cingulatus. Sharma etal . (2010) reported that the 1.0% concentration of A. indica (Neem Seed Kernel) causedabout 75% mortality in D. cingulatus. In conclusion, it can be stated that among the threesolvent extracts of C. scalpelliformis the chloroform extract has potential at sub lethalconcentration followed by the methanol and hexane extracts.

Antimicrobial activity:The hexane, chloroform and methanol extracts of C. scalpelliformis, C. veravalensis

and methanol extract of U. lactuca and U. fasciata inhibited the growth of F. oxysporum

(Tables 3). However, P. pavonica and S. wightii showed no activity against F. oxysporum.Alam et al. (2002) showed that different parts of Vinca rosea and Azadirachta indica showed

potential effect against F. oxysporum f.sp. vasinfectum. Suwitchayanon and Kunasakdakul etal. (2009) reported that the clove extract at the concentration of 2600 ppm was required forMIC to control Fusarium oxysoporum, in our experiment the C. scalpelliformis extract at theconcentration of 8mg/ml was required to inhibit this pathogen. Afifah (2010) investigated theantifungal activity of Halimeda discoidea and they reported that the algae was found toinhibit phytopathogenic fungus such as Aspergillus niger , Penicillium sp. and Rhizopus sp.Obongoya et al. (2010) investigated water based crude plant extracts of Neem ( Azadirachtaindica), Mexican marigold ( Tagetes minuta ) , tobacco ( Nicotiana tobacum ) and peri-winkle

(Vinca rosea ) in controlling soil-borne fungi (Fusarium oxysporum Schl. f. sp. phaseoli) ofcommon bean ( Phaseolus vulgaris L.). They found that Neem extract was the most effective,while Peri-winkle was the least in inhibiting F. oxysporum. The radial growth of Fusariumoxysporum f. sp. psidii was significantly less in neem leaf extract treatment followed

by Lantana leaf extract (Srivastava et al., 2011) treatments.

Liquid nanoparticle synthesis :Using extra cellular synthesis technique, we were able to produce physically stable

liquid nanoparticle formulation, both empty and seaweed thallus extract loaded. The changein color of empty and P. pavonica loaded liquid nanoparticle was noted by visual

observation. The stability of the P. pavonica loaded liquid nanoparticle (PPLNP) checked at16 hours, 24 hours and 16 weeks after synthesis by UV-vis spectroscopy. The UV-vis spectrarecorded from the PPLNP at different times of reaction are plotted. The strong surface

plasmon resonance centered at 422 nm clearly indicated an increase in intensity with time andstabilized after 5 minutes to 24 h of reaction. The metal particles were observed to be stablein PPLNP formulation from eight to twelve months after synthesis (Table 4) (still they arestable). Long-term stability of this formulation at room temperature indicating that there wasno observable variation in the optical properties of the nanoparticle solutions with time.


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Invariably all the bio-nanoparticles synthesized using marine algal seaweeds were sphericaland polydispersed and their size was ranged from 35.68 to 97.02 nm (Table 4). Similarlysilver nanoparticles synthesized using many plant extracts (Sahayaraj and Rajesh, 2011) arespherical and polydispersed as observed for marine algal seaweeds.

Secondary metabolites:Tannins, total phenols, ortho - dihydric phenols, bound phenols, alkaloids and flavanoidsquantities were estimated. Steroids, saponins and xanthoprotein can be measured at 480,420-500 and 500 nm respectively. Ahmad et al. (1994) reported that sterols are the commonsecondary metabolites of the brown algae. Tannin content was high in Sargassum wightii(61.20.2 g /g) followed by Caulerpa veravalensis (30.80.2 g /g). Minimum quantity has

been observed in Ulva lactuca (7.50.2 g /g). Total phenolic compound was high inCaulerpa veravalensis (12.20.1 mg/g), whereas, bound phenol was found maximum inSargassum wightii (7.90.1 g/g) and Ortho-di-hydric phenols in Caulerpa scalpelliformis(14.00.1 g/g) . Total flavanoids content was high in Ulva lactuca (14.70.2 mg/g) followed

by Sargassum wightii (14.10.4 mg/g) and minimum in Padina pavonica (11.50.3 mg/g).Distribution of secondary metabolites of marine algae is depends upon the season and habitat(Bhakuni and Rawat, 2005). However, in addtion to the season or habitate, type of algaealso place an important role for the distribution of phytochemicals. Moreover, thesemetabolities govern the economic importance of the marine algae (Cardozo et al., 2007).

Acknowledgements:The authors KS and JMR are grateful to MoES (Ref No. MRDF/01/33/P/07), Govt. of

India for the financial support for this research works. The authors are thankful to Dr.Eswaran, Scientist In charge, Central Salt and Marine Algal Research Station, Mandapam, foraiding in the identification of algae. We are also thankful to the management of St. XaviersCollege for the laboratory facilities and encouragement.

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In: Science against microbial pathogens: communicating currentresearch and technological advances (A. Mendez-Vilas Ed.), Volume 3, FORMATEX

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vitro Evaluation of Carbendazim 50% WP, Antagonists and BotanicalsAgainst Fusarium oxysporum f. sp. psidiiAssociated with Rhizosphere Soil of Guava. As ian Jo ur na l of Pl an t Pa th ol ogy , 5 (1): 46-53.Subba Rao, P.V. and V.A. Mantri. 2006. Indian seaweed resources and sustainableutilization: Scenario at the dawn of a new century. Current Science 91: 2, 164-174.Suwitchayanon, P. and Kunasakdakul, K. 2009. In Vitro effects of clove and turmeric extractscontrolling crucifer pathogens. J. Agri. Tech . 5(1): 193-199.Wang, B., Brubaker, C.L. and Burdon, J. 2004. Fusarium species and Fusarium Wilt

pathogens associated with native Gossypium populations in Austr


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Table 1. Algal seaweeds genus and species collected from various places of Kanyakumari,Tirunelveli, and Tuticorin and Ramanathapuram districts of Tamil Nadu

District LocationLatitude and

LongitudeNumber of Genus

Number of species

Kanyakumari

Manavalakurichi N 0808'30.8''

E 07718'09.7''

3 (8) 3 (6)

Kadiapatinam N 0807'46.6''E 07718'23.7''

3 (8) 4 (8)

Muttam N 0807'27.5''E 07718'48.8''

5 (13) 7 (13)

Kanyakumari N 0804'39.8''E 07733'01.8''

15 (38) 21 (39)

Circular Fort &Leepuram

N 0807'37.9''E 07734'02.7''

13(33) 18(34)

Tirunelveli

Kootapuli N 0808'44.2''E 07736'02.5''

6(11) 7(9)

Idhinthakarai N 0810'32.3''E 07744'31.3''

29(55) 48(59)

Kuthankuli N 0812'49.0''E 07746'58.4''

16(30) 23(29)

Uvari N 0817'05.1''E 07754'01.0''

2(4) 2(3)

Tuticorin

Manapaad N 0822'28.5''E 07703'54.6''

17(31) 26(35)

Tiruchendur N 0829'48.4''E 07807'47.8''

6(11) 8(11)

Tuticorin-Harbour &Hare island

N 0846'32.1''E 07811'56.5''

18(32) 25(34)

Therkukalmaedu N 0856'37.5''E 07811'55.0''

7(13) 8(11)

Jalli island N 0902'49.4''

E 07812'57.4''4(7) 4(5)

Keelvaipaar N 0902'31.4''E 07812'54.6''

1(2) 1(1)

Vaembaar N 0904'31.4''E 07821'50.3''

2(4) 2(3)

Ramanathapuram

Kizhakarai N 0902'31.4''E 07812'54.6''

3(17) 3(14)

Mandapam N 0904'31.4''E 07821'50.3''

11(61) 15(68)

Rameshwaram N 0913'34.7''E 07847'03.2'' 4(22) 4(18)

Value in parentheses indicates percentage with total population within the districtFigure 1. Total number of species of marine algae belonging to different groups occurring atfour districts of south Tamil Nadu Coast


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The SciTech Publishers & International Association or Teachin and Learnin , 2012

Table 2. Impact of selected algal seaweeds (hexane, chloroform, methanol and aqueousextracts) on LC 50 values and fiducidal limits, chi square parameters of D. cingulatus thirdinstar nymphs

Solvent LC 30 LC 50 LC 90 RegressionCoefficient

InterceptChi

Squaredf p

Caulerpa scalpelliformis

Hexane 136.58 271.05 1447.17 1.7617 -4.2862 3.013 3 0.390

Chloroform 51.64 108.23 660.31 1.6317 -3.3195 0.076 3 0.995

Methanol 153.93 275.43 1141.73 2.0752 -5.0635 9.322 3 0.025

Aqueous 3846.33 13127.22 263663.35 0.9836 -4.0508 1.945 3 0.584

Caulerpa veravalensis

Hexane 287.26 529.60 2361.62 1.9739 -5.3767 5.852 3 0.119

Chloroform 24.75 59.37 503.43 1.3804 -2.4483 1.864 3 0.601

Methanol 188.38 305.46 995.30 2.4982 -6.2078 11.393 3 0.010

Aqueous 1089.81 2294.26 14148.64 1.6221 -5.4512 2.665 3 0.446

Ulva fasciata

Hexane 400.91 875.13 5896.64 105468 -4.5507 4.131 3 0.248

Chloroform 244.87 493.96 2744.54 1.7207 -4.6351 2.798 3 0.424

Methanol 173.65 313.59 1329.46 2.0429 -5.0998 7.182 3 0.066

Aqueous 954.09 2282.47 19238.67 1.3843 -4.6491 0.233 3 0.972

Ulva lactuca

Chloroform 294.22 643.61 4359.48 1.5425 -4.3324 4.911 3 0.178

Methanol 170.89 399.27 3176.53 1.4229 -3.7012 2.224 3 0.527

Aqueous 1456.28 2938.49 16338.05 1.7200 -5.9653 1.488 3 0.685

Padina pavonica

7

18

11

46

98

4

14

21

15

7

0

5

10

15

20

25

Kanyakuma ri Ti runelveli Tuti cori n Ramanathapuram

N u m b e r o f

S p e c

i e s

Chlorophyta Paheophyta Rhodophyta


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Hexane 473.09 1326.45 16478.35 1.1712 -3.6573 2.001 3 0.572

Chloroform 354.30 1062.49 15556.72 1.0995 -3.3274 2.414 3 0.491

Methanol 448.26 1553.43 32388.28 0.9715 -3.1005 3.759 3 0.289

Aqueous 1150.01 2486.31 16363.67 1.5661 -5.3177 3.206 3 0.361

Sargassum wightii

Hexane 591.78 1439.09 12624.92 1.3588 -4.2913 1.103 3 0.776

Chloroform 311.78 631.79 3549.51 1.7097 -4.7881 1.456 3 0.692

Methanol 420.38 954.45 7080.22 1.4726 -4.3878 7.356 3 0.061

Aqueous 2089.98 4520.77 29789.71 1.5651 -5.7206 3.437 3 0.329

Table 3. Antifungal activity (Zone of inhibition in mm) of chosen sea weeds against Fusarium oxysporum f.sp . vasinfectum (n = 3)

Concentration

(%)Hexane Chloroform Methanol

Ulva lactuca

0.05 - - 6.00.6

0.1 - 5.30.3 8.00.5

0.2 - 5.30.6 10.30.3

0.4 - 6.30.1 11.30.2

0.8 - 8.70.3 11.70.1

Ulva fasciata

0.05 5.30.3 - 5.60.3

0.1 5.60.3 5.70.3 5.60.3

0.2 5.60.3 6.30.6 6.30.7

0.4 8.30.7 6.30.3 10.60.3

0.8 9.01.1 7.30.3 13.30.3

Positive control 18.60.3

- Indicates no activity recorded, MeanSE; Positive control 0.03% Carbendazim(Bavistin); Negative control Dimethyl Sulphoxide (DMSO)


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Table 4. Properties of biologically synthesized silver nanoparticles using selected marinealgae

Algal based silvernanoparticles

Particle shape Particle size Stability

Caulerpa veravalensis Spherical and polydispersed 44.421.75 08 months

Caulerpa scalpelliformis Spherical and polydispersed 58.633.91 08 months

Ulva fasciata Spherical and polydispersed 40.102.21 08 months

Ulva lactuca Spherical and polydispersed 35.681.16 08 months

Padina pavonica Spherical and polydispersed 45.734.20 24 months

Sargassum wightii Spherical and polydispersed 97.026.00 08 months

marine algae for the cotton pest and disease management

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