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List of Editors of Editors in the Journal of Research in Biology
Managing and Executive Editor:
Abiya Chelliah [Molecular Biology] Publisher, Journal of Research in Biology.
Editorial Board Members:
Ciccarese [Molecular Biology] Universita di Bari, Italy.
Sathishkumar [Plant Biotechnologist] Bharathiar University.
SUGANTHY [Entomologist] TNAU, Coimbatore.
Elanchezhyan [Agriculture, Entomology] TNAU, Tirunelveli.
Syed Mohsen Hosseini [Forestry & Ecology] Tarbiat Modares University (TMU), Iran.
Dr. Ramesh. C. K [Plant Tissue Culture] Sahyadri Science College, Karnataka.
Kamal Prasad Acharya [Conservation Biology] Norwegian University of Science and Technology (NTNU), Norway.
Dr. Ajay Singh [Zoology] Gorakhpur University, Gorakhpur
Dr. T. P. Mall [Ethnobotany and Plant pathoilogy] Kisan PG College, BAHRAICH
Ramesh Chandra [Hydrobiology, Zoology] S.S.(P.G.)College, Shahjahanpur, India.
Adarsh Pandey [Mycology and Plant Pathology] SS P.G.College, Shahjahanpur, India
Hanan El-Sayed Mohamed Abd El-All Osman [Plant Ecology] Al-Azhar university, Egypt
Ganga suresh [Microbiology] Sri Ram Nallamani Yadava College of Arts & Sciences, Tenkasi, India.
T.P. Mall [Ethnobotany, Plant pathology] Kisan PG College,BAHRAICH, India.
Mirza Hasanuzzaman [Agronomy, Weeds, Plant] Sher-e-Bangla Agricultural University, Bangladesh
Mukesh Kumar Chaubey [Immunology, Zoology] Mahatma Gandhi Post Graduate College, Gorakhpur, India.
N.K. Patel [Plant physiology & Ethno Botany] Sheth M.N.Science College, Patan, India.
Kumudben Babulal Patel [Bird, Ecology] Gujarat, India.
CHANDRAMOHAN [Biochemist] College of Applied Medical Sciences, King Saud University.
B.C. Behera [Natural product and their Bioprospecting] Agharkar Research Institute, Pune, INDIA.
Kuvalekar Aniket Arun [Biotechnology] Lecturer, Pune.
Mohd. Kamil Usmani [Entomology, Insect taxonomy] Aligarh Muslim university, Aligarh, india.
Dr. Lachhman Das Singla [Veterinary Parasitology] Guru Angad Dev Veterinary and Animal Sciences University, Ludhiana, India.
Vaclav Vetvicka [Immunomodulators and Breast Cancer] University of Louisville, Kentucky.
José F. González-Maya [Conservation Biology] Laboratorio de ecología y conservación de fauna Silvestre, Instituto de Ecología, UNAM, México.
Dr. Afreenish Hassan [Microbiology] Department of Pathology, Army Medical College, Rawalpindi, Pakistan.
Gurjit Singh [Soil Science] Krishi Vigyan Kendra, Amritsar, Punjab, India.
Dr. Marcela Pagano [Mycology] Universidade Federal de São João del-Rei, Brazil.
Dr.Amit Baran Sharangi [Horticulture] BCKV (Agri University), West Bengal, INDIA.
Dr. Bhargava [Melittopalynology] School of Chemical & Biotechnology, Sastra University, Tamilnadu, INDIA.
Dr. Sri Lakshmi Sunitha Merla [Plant Biotechnology] Jawaharlal Technological University, Hyderabad.
Dr. Mrs. Kaiser Jamil [Biotechnology] Bhagwan Mahavir Medical Research Centre, Hyderabad, India.
Ahmed Mohammed El Naim [Agronomy] University of Kordofan, Elobeid-SUDAN.
Dr. Zohair Rahemo [Parasitology] University of Mosul, Mosul,Iraq.
Dr. Birendra Kumar [Breeding and Genetic improvement] Central Institute of Medicinal and Aromatic Plants, Lucknow, India.
Dr. Sanjay M. Dave [Ornithology and Ecology] Hem. North Gujarat University, Patan.
Dr. Nand Lal [Micropropagation Technology Development] C.S.J.M. University, India.
Fábio M. da Costa [Biotechnology: Integrated pest control, genetics] Federal University of Rondônia, Brazil.
Marcel Avramiuc [Biologist] Stefan cel Mare University of Suceava, Romania.
Dr. Meera Srivastava [Hematology , Entomology] Govt. Dungar College, Bikaner.
P. Gurusaravanan [Plant Biology ,Plant Biotechnology and Plant Science] School of Life Sciences, Bharathidasan University, India.
Dr. Mrs Kavita Sharma [Botany] Arts and commerce girl’s college Raipur (C.G.), India.
Suwattana Pruksasri [Enzyme technology, Biochemical Engineering] Silpakorn University, Thailand.
Dr.Vishwas Balasaheb Sakhare [Reservoir Fisheries] Yogeshwari Mahavidyalaya, Ambajogai, India.
Dr. Pankaj Sah [Environmental Science, Plant Ecology] Higher College of Technology (HCT), Al-Khuwair.
Dr. Erkan Kalipci [Environmental Engineering] Selcuk University, Turkey.
Dr Gajendra Pandurang Jagtap [Plant Pathology] College of Agriculture, India.
Dr. Arun M. Chilke [Biochemistry, Enzymology, Histochemistry] Shree Shivaji Arts, Commerce & Science College, India.
Dr. AC. Tangavelou [Biodiversity, Plant Taxonomy] Bio-Science Research Foundation, India.
Nasroallah Moradi Kor [Animal Science] Razi University of Agricultural Sciences and Natural Resources, Iran
T. Badal Singh [plant tissue culture] Panjab University, India
Dr. Kalyan Chakraborti [Agriculture, Pomology, horticulture] AICRP on Sub-Tropical Fruits, Bidhan Chandra Krishi Viswavidyalaya,
Kalyani, Nadia, West Bengal, India.
Dr. Monanjali Bandyopadhyay [Farmlore, Traditional and indigenous
practices, Ethno botany] V. C., Vidyasagar University, Midnapore.
M.Sugumaran [Phytochemistry] Adhiparasakthi College of Pharmacy, Melmaruvathur, Kancheepuram District.
Prashanth N S [Public health, Medicine] Institute of Public Health, Bangalore.
Tariq Aftab Department of Botany, Aligarh Muslim University, Aligarh, India.
Manzoor Ahmad Shah Department of Botany, University of Kashmir, Srinagar, India.
Syampungani Stephen School of Natural Resources, Copperbelt University, Kitwe, Zambia.
Iheanyi Omezuruike OKONKO Department of Biochemistry & Microbiology, Lead City University,
Ibadan, Nigeria.
Sharangouda Patil Toxicology Laboratory, Bioenergetics & Environmental Sciences Division,
National Institue of Animal Nutrition
and Physiology (NIANP, ICAR), Adugodi, Bangalore.
Jayapal Nandyal, Kurnool, Andrapradesh, India.
T.S. Pathan [Aquatic toxicology and Fish biology] Department of Zoology, Kalikadevi Senior College, Shirur, India.
Aparna Sarkar [Physiology and biochemistry] Amity Institute of Physiotherapy, Amity campus, Noida, INDIA.
Dr. Amit Bandyopadhyay [Sports & Exercise Physiology] Department of Physiology, University of Calcutta, Kolkata, INDIA .
Maruthi [Plant Biotechnology] Dept of Biotechnology, SDM College (Autonomous),
Ujire Dakshina Kannada, India.
Veeranna [Biotechnology] Dept of Biotechnology, SDM College (Autonomous), Ujire Dakshina Kannada, India.
RAVI [Biotechnology & Bioinformatics] Department of Botany, Government Arts College, Coimbatore, India.
Sadanand Mallappa Yamakanamardi [Zoology] Department of Zoology, University of Mysore, Mysore, India.
Anoop Das [Ornithologist] Research Department of Zoology, MES Mampad College, Kerala, India.
Dr. Satish Ambadas Bhalerao [Environmental Botany] Wilson College, Mumbai
Rafael Gomez Kosky [Plant Biotechnology] Instituto de Biotecnología de las Plantas, Universidad Central de Las Villas
Eudriano Costa [Aquatic Bioecology] IOUSP - Instituto Oceanográfico da Universidade de São Paulo, Brasil
M. Bubesh Guptha [Wildlife Biologist] Wildlife Management Circle (WLMC), India
Rajib Roychowdhury [Plant science] Centre for biotechnology visva-bharati, India.
Dr. S.M.Gopinath [Environmental Biotechnology] Acharya Institute of Technology, Bangalore.
Dr. U.S. Mahadeva Rao [Bio Chemistry] Universiti Sultan Zainal Abidin, Malaysia.
Hérida Regina Nunes Salgado [Pharmacist] Unesp - Universidade Estadual Paulista, Brazil
Mandava Venkata Basaveswara Rao [Chemistry] Krishna University, India.
Dr. Mostafa Mohamed Rady [Agricultural Sciences] Fayoum University, Egypt.
Dr. Hazim Jabbar Shah Ali [Poultry Science] College of Agriculture, University of Baghdad , Iraq.
Danial Kahrizi [Plant Biotechnology, Plant Breeding,Genetics]
Agronomy and Plant Breeding Dept., Razi University, Iran
Dr. Houhun LI [Systematics of Microlepidoptera, Zoogeography, Coevolution,
Forest protection] College of Life Sciences, Nankai University, China.
María de la Concepción García Aguilar [Biology] Center for Scientific Research and Higher Education of Ensenada, B. C., Mexico
Fernando Reboredo [Archaeobotany, Forestry, Ecophysiology] New University of Lisbon, Caparica, Portugal
Dr. Pritam Chattopadhyay [Agricultural Biotech, Food Biotech, Plant Biotech] Visva-Bharati (a Central University), India
Table of Contents (Volume 4 - Issue 4)
Serial No Accession No Title of the article Page No
1 RA0446 Laboratory evaluation and comparative study of herbal mosquito coils
against the filarial vector, Culex quinquefasciatus (Diptera: Culicidae).
Susheela P and Radha R.
1332-1337
2 RA0447 Daily Activity Budget of Nicobar Long-tailed Macaque (Macaca
fascicularis umbrosa) in Great Nicobar Island, India..
Rajeshkumar S, Raghunathan C, Kailash Chandra and Venkataraman K.
1338-1347
3
RA0454
Analysis on protein fingerprint, RAPD and fruit quality of tomato
mutants by ion beam implantation.
Duan HY, Wang CF, Yu YA, Li XW and Zhou YQ.
1348-1356
4
RA0452
The leaping behavior of the sally lightfoot crab Grapsus grapsus
(Crustacea: Decapoda: Brachyura) at an oceanic archipelago.
Marina de Sá Leitão Câmara de Araújo.
1357-1364
Article Citation: Susheela P and Radha R. Laboratory evaluation and comparative study of herbal mosquito coils against the filarial vector, Culex quinquefasciatus (Diptera: Culicidae) Journal of Research in Biology (2014) 4(4): 1332-1337
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Laboratory evaluation and comparative study of herbal mosquito coils
against the filarial vector, Culex quinquefasciatus (Diptera: Culicidae)
Keywords: Mosquito, Culex quinquefasciatus, repellency, Plant essential oil.
ABSTRACT: Synthetic insecticides employed for the control of insect pests are toxic to man and livestock acting as pollutants to the environment, killing all beneficial insects thereby causing a disturbance to the ecosystem. The use of natural products such as plant essential oils has assumed significance as an important component of insect pest management because of their financial viability and eco-friendly nature. They hold promise as alternatives to chemical insecticides to reduce pesticide load in the environment. A laboratory experiment was conducted to investigate the efficacy of three essential oils -eucalyptus oil, lemon grass oil and thyme oil for the repellent activity against the filarial vector, Culex quinquefasciatus. Among the essential oils, Lemon grass oil showed good repellency property when compared to the other two plant oils. Hence, the results of the investigation would indicate a significant potential for lemon grass oil as a possible source of natural products that could be used as an alternative to synthetic insecticides.
1332-1337 | JRB | 2014 | Vol 4 | No 4
This article is governed by the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.0), which gives permission for unrestricted use, non-commercial, distribution and reproduction in all medium, provided the original work is properly cited.
www.jresearchbiology.com Journal of Research in Biology
An International
Scientific Research Journal
Authors:
Susheela P* and Radha R.
Institution:
Department of Zoology,
PSGR Krishnammal College
for Women Coimbatore,
Tamilnadu, India.
Corresponding author:
Susheela P.
Web Address: http://jresearchbiology.com/
documents/RA0446.pdf.
Dates: Received: 01 April 2014 Accepted: 31 May 2014 Published: 20 Jun 2014
Journal of Research in Biology
An International Scientific Research Journal
Original Research
ISSN No: Print: 2231 –6280; Online: 2231- 6299
INTRODUCTION
Mosquitoes are considered as an important insect
pests that affect the health and well being of human
beings and other animals worldwide. Mosquitoes are
cosmopolitan in distribution and have occupied many
niches including higher altitudes. Mosquitoes are always
considered as a nuisance because they consume blood
from living vertebrates, including human beings
(Bernhard et al., 2003). In India, annually around 40
million people suffer from mosquito borne diseases. The
extensive use of mosquito repellents and insecticides in
public health programmes has caused extensive level of
environmental pollution and serious health hazards.
Many of them are alarmingly toxic to human beings and
also other non-target organisms.
Controlling the mosquitoes in an effective manner is
often complex and expensive task which requires support
from communities and also from different groups such as
industry, agriculture, state and local governments
(Joseph et al., 2004). The harmful effect of the pesticides
on the environment, animals, plants and human beings is
an issue of great concern. As far as India is concerned,
many of the insecticides and larvicides are
commercialized in the form of dust, powder or sprays
that contain chemicals such as organochlorine,
organophosphates and synthetic pyrethroid. Yet
mosquitoes, due to a prolonged use of these insecticides
become resistant and thus it becomes a difficult task to
eradicate them totally (Prajapati et al., 2005). They also
pose a threat to the human population by carrying vector
borne diseases and sometimes out break as epidemics.
Hence to control the vector mosquitoes, efforts are
being taken to look for an alternate solution which
will ultimately minimize the use of synthetic
insecticides.
The development of eco-friendly insecticides will
serve its purpose as a new alternate to substitute the
synthetic insecticides essentially cutting down the
chemical pollution. The pyrethrum flower extracts
contain active materials that are potential enough to
control the mosquito population. (Sutthanont et al.,
2010). In recent times, plant products are used as novel
chemo therapeutants in pest management in different
parts of the world, because of their biodegradable nature.
(Hardin and Jackson, 2009).Therefore, the present study
was aimed to investigate the mosquito repellent nature of
three essential oils: Eucalyptus tereticornis (Eucalyptus),
Cymbopogon citratus (lemon grass) and Thymus vulgaris
(thyme) against C. quinquefasciatus.
MATERIAL AND METHODS
Plant Oils:
The plant oils were purchased from the Aromatic
Oil Stores, Coimbatore, Tamil Nadu and formulated for
the experiment. A stock solution at 1000 ppm is prepared
by dissolving the essential oils in distilled water using
2 ml of 100% acetone respectively. The serial dilutions
of essential oils at the concentration of 5%, 15% and
25% and three replicate of each concentration were
made.
Preparation of herbal mosquito coils:
Mosquito coils were prepared using cow dung,
sawdust, neem leaves, flower waste and tulsi leaves.
Then the essential oils, Thymus vulgaris, Lemon grass,
and Eucalyptus oils were sprayed (w/w) on top of the
coil by using a hand spray pump in different
concentration of 5%, 15% and 25 % separately and they
were used for its efficacy against C. quinquefasciatus
mosquito. The coil was dried in the oven at 70°C for
6 hours was dried for half an hour at room temperature.
These coils were then packed in suitable air tight plastic
folders and kept for 2 – 3 days for even spread of the
essential herbals on the coil.
Test Organisms
The test organism, C. quinquefasciatus, was reared in
the laboratory in the Department of Zoology, PSGR
Krishnammal College for Women, Coimbatore, Tamil
Nadu. Dog biscuits and yeast powder in a ratio of 3:1
Susheela and Radha, 2014
1333 Journal of Research in Biology (2014) 4(4): 1332-1337
were given as feed for the mosquito larvae. On the other
hand, adult mosquitoes were fed with a 10% sucrose
solution and a 1 week-old chick. Mosquitoes were kept
at relative humidity of 28-30°C, 75 ± 5%, with 14-h light
and 10-h dark, photo period ( Kitzmiller et al., 1954).
Bioassays
Repellency Test
The experiment was conducted in a closed room,
with a volume of 92.8 m3 in the Department of Zoology,
PSGR Krishnammal College for Women, Coimbatore,
Tamil Nadu. The human volunteers sat at 1 m, 2 m, 4 m,
and 8 m from the herbal mosquito coil. The mosquito
coil was put in the middle of one side of the room. For
control, 50 female unfed, 5 days old mosquitoes were
released in the centre of the room. Then the number of
landing mosquitoes on the bare legs of the human
volunteers was counted for a period of 2 min. For testing,
the mosquito coil was ignited, then counting of the
number of landing mosquitoes on the bare legs of the
human volunteers began and was recorded at periodic
intervals. Three replications were done by changing the
positions of the human volunteers, and then repeating the
procedure the next day.
Journal of Research in Biology (2014) 4(4): 1332-1337 1334
Susheela and Radha, 2014
Figure-1 Repellency of lemon grass oil against C. quinquefasciatus
Concentration of lemon grass oil
Concentration of eucalyptus oil
Figure-2 Repellency of eucalypus oil against C. quinquefasciatus
RESULTS AND DISCUSSION
The results of repellency test of thyme oil against
C. quinquefasciatus (Say) after one hour of treatment are
presented in Figure-3. The results clearly indicated that
the highest repellency was reported at 25% concentration
of thyme oil when compared to 5% concentration and
10% concentration. As the concentration of the plant oil
formulation increases the total mortality of
C. quinquefasciatus also gets increased. Figure-2
revealed the efficacy of eucalyptus oil against
C. quinquefasciatus. The lowest repellency was observed
at 5% concentration of eucalypus oil and the highest
repellency was observed at 25% concentration. But the
essential oil, eucalypus oil is more effective than thyme
oil. Increase in the concentration of the plant oil
formulation was found to increase the total repellency of
Culex quinquefasciatus. The different concentrations of
the lemon grass oil was recorded against
Culex quinquefasciatus in Figure-1. The percentage of
repellency was found to be high in 25 % concentration
than 5 % concentration of the plant oil. The results of
this study clearly indicated that lemon grass oil had high
repellency potential to control the mosquitoes than the
other two essential oils.
A number of studies have been focused on lemon
grass oil for controlling mosquitoes as a larvicide and a
repellent with varied results. Hanifah et al., (2011)
demonstrated C. citratus extract has more acaricidal
activity against Dermatophagoides farina and
D. pteronyssinus than Azadirachta indica at 50%
concentration. This proves the efficiency of
Cymbopogon citratus in controlling the insect pests.
Oyedele et al., (2002) evaluated the ointment and cream
formulations of lemon grass oil in different classes of
base and the oil in liquid paraffin solution for mosquito
repellency in a topical application. Cilek et al., (2011)
studied the efficacy of several commercially formulated
essential oils against caged female Aedes albopictus and
Culex quinquefasciatus. Mgbemena (2010) found that
the essential oil O. gratissimium had a greater larvicidal
activity than C. citratus. Purwal et al., (2010) tested the
activity of C. citratus and Mentha piperita essential oils
in a combination against Pediculus humanus and found
a mean time to death of 60 minutes. Therefore the
essential oils can be used as an alternative to synthetic
insecticides for vector control programmes.
The essential oils (EO) eucalyptus oil, lemon
grass oil, thyme oil were evaluated for repellent activity
against the Culex quinquefasciatus. Essential oils of
Concentration of thyme oil
Figure-3 Repellency of thyme oil against C. quinquefasciatus
Susheela and Radha, 2014
1335 Journal of Research in Biology (2014) 4(4): 1332-1337
many plants were observed to have mosquito larvicidal
property and have received attention as potentially
controlling vectors of mosquito borne disease (Zhu et al.,
2006). Therefore, the use of lemon grass oils in insect/
mosquito control is an alternative pest control method for
minimizing the harmful effects of pesticidal compounds
on the environment. The present study has identified
more plant oils showing larvicidal activity against
Culex mosquito. The results obtained suggest that the
plant oils are promising as larvicides against
Culex mosquito. The present study also suggests the use
of Lemon grass oil as the most effective alternative in
controlling mosquitoes.
CONCLUSION
The results of the present investigation proved
that the all essential oils at higher concentration are
effective but lemongrass oil exhibit a significant knock
down activity at higher concentration when compared to
the other oils. For the commercialization of these herbal
mosquito coils, further simulated and actual field trials
are required. Hence, Lemongrass essential oil, alone or in
combinations with those obtained from other mosquito
repellent plant species, could be potentially used for the
preparation of mosquito repellent products.
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1337 Journal of Research in Biology (2014) 4(4): 1332-1337
Article Citation: Rajeshkumar S, Raghunathan C, Kailash Chandra and Venkataraman K. Daily Activity Budget of Nicobar Long-tailed Macaque (Macaca fascicularis umbrosa) in Great Nicobar Island, India. Journal of Research in Biology (2014) 4(4): 1338-1347
Jou
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Daily Activity Budget of Nicobar Long-tailed Macaque
(Macaca fascicularis umbrosa) in Great Nicobar Island, India
Keywords: Macaca fascicularis umbrosa, Daily activity budget, Great Nicobar Island
ABSTRACT: Nicobar long-tailed macaques (Macaca fascicularis umbrosa Miller, 1902) are distributed in three Islands of Nicobar namely Great Nicobar, Little Nicobar and Katchal. Their insular population requires special attention from research and management perspectives. Daily activity budget of M.f. umbrosa in the Great Nicobar Island was studied from October 2011 to September 2013 by intensive direct observation method. Study revealed that Nicobar long-tailed macaque, undergoes most of the time for Locomotion (36.07%), followed by feeding (22.35%), resting or being inactive (15.74%), grooming (11.14%), vocalization (7.03%), playing (5.64%), sexual arousal (1.46%) and agonistic (0.56%). All daily activities have significant difference (χ2 = 1156.22; df = 7, P = 0.05). Chi-square test demonstrated that the daily activity budget differed significantly among the behaviours. Qualitative results found that the interaction within the group was fighting and grabbing food. The significant observation of disability in their legs was noticed in Nicobar Long-tailed Macaque. The relation between their behaviour and disability is also discussed.
1338-1347 | JRB | 2014 | Vol 4 | No 4
This article is governed by the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.0), which gives permission for unrestricted use, non-commercial, distribution and reproduction in all medium, provided the original work is properly cited.
www.jresearchbiology.com Journal of Research in Biology
An International
Scientific Research Journal
Authors:
Rajeshkumar S1*,
Raghunathan C1,
Kailash Chandra2 and
Venkataraman K2.
Institution:
1. Zoological Survey of
India, Andaman and Nicobar
Regional Centre, Port Blair-
744 102, Andaman and
Nicobar Islands, India.
2. Zoological Survey of
India, M-Block, New
Alipore, Kolkatta-700 053,
India.
Corresponding author:
Rajeshkumar S.
Email Id:
Web Address: http://jresearchbiology.com/
documents/RA0447.pdf.
Dates: Received: 01 Apr 2014 Accepted: 30 May 2014 Published: 24 Jun 2014
Journal of Research in Biology
An International Scientific Research Journal
Original Research
ISSN No: Print: 2231 –6280; Online: 2231- 6299
INTRODUCTION
Primates are maintaining the sustainable
ecosystem and play as indicator for ecosystem health;
hence, they help in making of conservation and
management plans. Non-human primates of undisturbed
areas are having great behavioural variation (Thomas,
1991) which are closely related to human beings such as
eating, playing, fighting, keeping young ones etc. (Rod
and Preston-Mafham, 1992). The daily activities and
behaviour of primates differ between residential, non-
residential and undisturbed areas (Krebs and Davies,
1993). Large group size, poor habitat quality, seasonal
variation in food availability may affect their daily
activity budget (Peres, 1993; Passamani, 1998). The
Long-tailed macaques (Macaca fascicularis umbrosa
Miller, 1902) are the only non-human primates found on
Nicobar Islands (Umapathy et al., 2003). Other
subspecies occur in Myanmar, Cambodia, Laos,
Vietnam, Thailand, Malaysia, Indonesia and the
Philippines (Rodman, 1991; Tikader and Das, 1985).
This species varies in their behaviour, social
organisations, habitat consumption, morphology and
genetic variation due to wide distribution (Brent and
Veira, 2002; Hamada et al., 2008). Previous researches
in Nicobar subspecies are available for population status
and distribution profiling (Umapathy et al., 2003;
Sivakumar, 2010; Narasimmarajan and Raghunathan,
2012) Study on ecology and behaviour are also focused
in the other subspecies of Long-tailed Macaque in South
East Asian countries. Reports are available on the
aggressive and social behaviour of M. fascicularis
(Nordin and Jasmi, 1981; Zamzarina, 2003; Brent and
Veira, 2002; Khor, 2003; Md-Zain et al., 2003; Siti,
2003). The present study is focused on the daily activity
budgets of M f. umbrosa in Great Nicobar Island.
MATERIALS AND METHODS
Study Area
The Great Nicobar Island is about 1045.1 sq km
comprises of Campbell bay National Park and Galathea
National Park (Fig. 1). These two National Parks
embrace Great Nicobar Biosphere Reserve (GNBR). The
study site covers about 3 km2 and is composed of low
hills near dense semi evergreen forest, Maggar Nallah
river and human Settlements at Govind Nagar (06°
59.985' N 093° 54.459' E) and it is 6 km away from
Campbell Bay (Fig. 1). GNBR has richest faunal and
Rajeshkumar et al., 2014
1339 Journal of Research in Biology (2014) 4(4): 1338-1347
Fig 1. Study area and Study site.
floral communities. Great Nicobar is the home for plants
like Albizia chinensis, Albizia lebbeck, Artocarpus
chaplasha, Calophyllum soulattri, Dipterocarpus sp.,
Pterocarpus sp., and Sterculia campanulatum. In fauna,
other than the long-tailed macaques, the endemic
mammals recorded are Nicobar wild boar (Sus scrofa
nicobaricus), Nicobar Tree shrew (Tupaia nicobarica),
Nicobar shrew (Crocidura nicobarica) and Nicobar
Flying fox (Pteropus faunulus).
Behaviour Sampling Method
Following the methods of Hambali et al., (2012);
Md-Zain et al., (2008b) and Brent and Veira (2002) daily
activity observations of macaque were made during 2 to
3 days in a week at 0500 hours until 1630 hours for 78
days from October 2011 to September 2013 to determine
the behaviour categories. A study group categories and
its composition of the three consecutive years are given
in Table. 1. The total number of individuals in study
group increased year by year i:e from 37 to 51
individuals. Every year the numbers of females were
more than that of males. This group was marked by their
dominant male who had a distinctive large and elongated
white area between the eyes and white eyelids compared
to the other groups. Focal animal sampling method was
adopted to collect the quantitative data at ten minutes
interval (Altmann, 1974; Lehner, 1979). During
torrential rain and adverse weather condition, the
observation was discontinued until the weather resumes
normally, because the animals were partially obscured or
moved completely from the observation sites. The data
on the observations of locomotion, feeding, resting,
grooming, vocalization, playing, sexual arousal and
agonistic were collected during the study. Chi-square test
was applied to analyse the behaviour data set obtained.
The nonparametric χ2 test was used to analyze the
significance of activity budgets.
RESULTS AND DISCUSSION
Result on the percentage of eight daily activities
of Nicobar long-tailed macaques monitored is given in
Table 2. Chi-square analysis upon the present study
indicated that all the eight behavioural observation shows
significant differences (Table 2). Jaman and Huffman
(2008) observed that, activities of Japanese macaque
(M. fuscata) in captivity varied between age-sex classes.
Similarly the behavioural variation occurred in
individual with different age-sex observed in the present
study. The most observed daily activity for all the age
group was locomotion. The locomotion is the highest
portion of daily activity in long-tailed macaques
compared to other activities (Hambali et al., 2012; Md-
Zain et al., 2010; Sia, 2004; Suhailan, 2004). This is
because of diurnal in nature as they are very active
during the day as they use their maximum time in
searching for food.
Locomotion
According to Menard (2004) and Wheatley
(1980) Long-tailed Macaques are the primates spending
most of their time for moving as they are mainly
frugivorous and occupy more space. It was also observed
that the study group’s moving choice is varied day by
day to different location and range. When they move out
Journal of Research in Biology (2014) 4(4): 1338-1347 1340
Rajeshkumar et al., 2014
Group categories Adult (Mature) Immature
Total No. of Individual
Year Male Female Total Sub Adult Juvenile Infant
2011 (October) 10 13 23 10 3 1 37
2012 (March) 12 15 27 12 4 2 45
2013 (August) 13 16 29 12 6 4 51
Table 1. Year wise group composition and total number of Individuals in the study group.
of their home range, there was a shortage of food sources
and availability of fruits. According to O’Brien and
Kinnaird (1997), availability of food source significantly
affects their locomotion in daily activity pattern.
Sometimes these animals visit human settlement areas
and raids crop land, coconuts farms and banana farms
which lead to their destruction. The result indicates that
the macaque spent most of the time in moving due to the
insufficient food sources in their habitat. Likewise this
study group also spend most of their time to visit
different localities because of their diminishing natural
food sources.
Feeding
Besides locomotion, feeding was observed as one
of the major activities of macaque during the study (Fig.
2 A). It resembles with the other subspecies studied by
Hambali et al., (2012), Md-Zain et al., (2010), Suhailan
(2004) and Tuan-Zaubidah (2003) who all found that
feeding is the second most occurrence activity compared
to other. However this finding was contradict with other
macaque species. For example Southern India wild lion-
tailed macaque (Kurup and Kumar, 1993) and captive
Japanese macaque (Jaman and Huffman, 2008) spend the
highest proportion of time in resting rather than feeding
depending on the food and weather factor. An increase in
one activity may pose some influence on other activities
(Jaman and Huffman, 2008). The main food sources are
fruits, flowers, tender leaves, insects, crabs, beetles,
butterflies, some spiders, grasshopper etc. Usually
macaque feed insects in afternoon period between resting
and grooming. When the food sources are less long-
tailed macaque usually rest.
Resting
Resting is the third most activity observed in our
study (Fig. 2 B). The result of the study revealed that
prolonged feeding activity considerably reduced the
resting behaviour during the observation from macaque
in Great Nicobar as noticed by Hambali et al., (2012) in
Malayan long-tailed macaque and Kurup and Kumar
(1993) in lion-tailed macaque. Resting includes activities
like sleeping, lying down and to sit idle. Macaques were
observed resting on tree branches, dead woods, bushes,
rocks and sometimes resting on the roads. Also they use
to take a few minutes rest after walking continuously.
Rainy season and unusual climate directly affect their
feeding and moving activities and increase their resting
Rajeshkumar et al., 2014
1341 Journal of Research in Biology (2014) 4(4): 1338-1347
Activity Observation Percentage (%) Expected frequency χ2 = (O-E)2/E
Locomotion 518 36.07 179.5 638.34*
Feeding 321 22.35 179.5 111.54*
Resting 226 15.74 179.5 12.04*
Grooming 160 11.14 179.5 2.12*
Vocalization 101 7.03 179.5 34.33*
Playing 81 5.64 179.5 54.05*
Sexual 21 1.46 179.5 139.95*
Agonistic 08 0.56 179.5 163.85*
Total 1436 100 1436 1156.25
Table 2. Percentage and Chi-square value of Nicobar long-tailed macaque’s daily activity.
* Showing significant differences (p<0.05), by using the chi-square test (χ2).
Degrees of freedom (df) = 7, O-Observation, E-Expected frequency.
activity. During night time, macaques sleep on the top of
tree branches. This behaviour indicates that the macaque
protect themselves from the predators. The only known
predator is reticulated python (Broghammerus
reticulatus) as no other higher predators are found in
Great Nicobar Island, but the anthropogenic activity and
domestic predators like dogs also affects their normal
activity.
Grooming
Grooming is the fourth highest activity observed
after resting (Fig. 2 C). This result is similar with M.
fascicularis found in Kuala Selangor Nature Park,
Malaysia (Hambali et al., 2012). Most of their grooming
activity occurs at the time of resting period. It was
predominantly observed at late afternoon when the
macaques return to the home range. At the time of
grooming one monkey picks up lice from other’s body.
Most of the individuals often prefer to self-groom rather
than social grooming. Social grooming highly noticed
between the adult female and adult male. Observations
on grooming between the adult female with infants were
least due to the presence of only few infant in the group.
There was a least observation on grooming between
adult female and juveniles as well as sub adults. Self-
grooming was also often observed in sub adults and
secluded male at the time of resting. In addition, after
mating, the dominant male is groomed by female.
According to Lazaro-perea et al., (2004) this behaviour
can be a way to get protection from others while fighting
and also for sharing of food.
Vocalization
Vocalization is the fifth behaviour that has been
observed. When the agonistic interaction occurs between
the group individuals, dominant adult male produce loud
calls and all the other individuals sound continuously. In
general, macaque produces loud calls especially for
Rajeshkumar et al., 2014
Journal of Research in Biology (2014) 4(4): 1338-1347 1342
Fig 2. Various activities of Long-tailed Macaque in Great Nicobar Island
A. Feeding, B. Resting, C. Grooming, D. Playing, E. Mating, F. Agonistic.
grabbing and snatching food item and fighting with their
group member. In addition during agonistic interaction
within the group or entrance of predatory animals such as
dogs in their territory, macaque used to make
vocalization. Normally vocalization can be treated as a
warning signal to protect themselves from predators as
observed by Md-Zain et al., (2010). Due to the
observer’s or the human’s activity in their range,
macaque produce different sounds and mainly the sub
adults seem to be most active as they used to climb very
quickly and keep other individuals alert. Members of the
group after hearing the vocal call warning used to climb
to higher ground to escape or hide in bushes. We
observed a least number of calls produced by macaques
while playing activity. Kipper and Todt (2002) and Md-
Zain et al., (2010) also found that the vocal call was
produced by macaques while playing. In the present
study the male long-tailed macaques were found to
produce vocal calls while grooming after mating. No
females were observed producing vocals during mating.
On the other hand observation made by Md-Zain et al.,
(2010) showed that females were found to produce vocal
during and after mating. The possible reason for this
behaviour can be a hormonal effect (Engelhardt et al.,
2005).
Playing
Playing activity is the sixth behaviour that has
been observed during the study period (Fig. 2 D). We
found predictable differences in playing activity in the
juveniles and sub adults. Juveniles were found to play
more than sub adults. Adult macaques were not involved
in playing activity. The playing behaviour may form a
social competition and juveniles in their active age
period will learn on social relations (Kipper and Todt,
2002). Usually, playing behaviour was observed in the
late afternoon, when adult long-tailed macaques are
inactive. Wrestling, chasing, tickling, swinging on the
tree branches, pulling their tails to play with one another
and invert hanging and jumping were the playing
categories observed during the study. It was also
observed that these animals prefer playing on the
selected trees like Casuarinas, Pandanus, Guava and
Coconut. In the evening, all the group member moves
near sleeping site and while moving many were found
collecting and eating some insects in the bushy area.
Sexual Arousal
Sexual behaviour like mating, mount, inspect
copulation are the categories were observed as the
seventh activity (Fig. 2 E). In our study period dominant
males were actively involved in mating with adult
females as this may help females in giving birth to
healthy generation. Females use to live with multimale
group, focused in copulating with dominant males as
observed by Hambali et al., (2012), Lawler et al., (1995),
Md-zain et al., (2010) and Van Noordwijk and Van
Schaik (1999). Sexual behaviour observed is only a small
portion of daily activity in long-tailed macaque.
Normally the adult male was found to smell or observe
the adult female genitalia first to make sure that the
females are ready to mate or not which is in corroborated
with the report of Brent and Veira (2002), Md Zain et al.,
(2010) and Hambali et al., (2012). The long-tailed
macaque takes a few seconds for mating activity.
Agonistic Activity
The least observed activity is the agonistic
behaviour (Fig. 2 F). During our study chase, grab, hit,
bite and fight are the categories of agonistic behaviour
observed as the eighth activity. Though these behaviours
are supported by Hambali et al., (2012), Md-Zain et al.,
(2010), Suhailan (2004) and Tuan-Zaubidah (2003) they
found that mating is the least observed activity. Fighting
behaviour occurred while gaining foods and mates.
Hambali et al., (2012) found that Malay wild long-tailed
macaque has a hierarchy in the group, so that they have
their own way to avoid fight when looking for food
together. Chasing and biting occur sometime between the
males and sub adults. Adult male were more aggressive
when their food was grabbed by other males, this shows
Rajeshkumar et al., 2014
1343 Journal of Research in Biology (2014) 4(4): 1338-1347
that the aggression appeared in males higher than
females which is agreed with the Brent and Veira (2002)
from macaque observed at Indo-China population.
Significantly we observed few aggressive activities in the
Nicobar long-tailed macaque against human beings
especially women and children during the study period.
Disability and Behaviour
During our study period several disabled
macaques were spotted (Fig 3). They were not able to
move properly due to their disability. These disabilities
may cause some changes in their daily activities which in
turn will cause changes in their behaviour like
locomotion, disability in finding mates, foraging
activities, etc. The relation between disability and
behaviour is also reported in Japanese macaques
(Macaca fuscata) by Turner et al., (2012). The possible
causes of disabilities are congenital defects, dog chasing
and anthropogenic activities. However, exact cause of
disability was not known. But this significant
observation may throw some light on the threats and
their status of these monkeys.
CONCLUSIONS
The present study enlightened behavioural and
activity patterns of the long-tailed macaque population
living in the Great Nicobar Island. It is revealed that
Rajeshkumar et al., 2014
Journal of Research in Biology (2014) 4(4): 1338-1347 1344
Fig 3. Disability in Nicobar Long-tailed macaque
A. Forearm partially disabled, B. Foreleg disabled, C. Hindleg partially disabled.
locomotion, feeding and resting were the most common
daily activities of these monkeys. Disabled macaques
spotted during our study period may give some
information on the changes in their behaviour that occur
due to disability as well as on the threats they use to
encounter. This study also found that the aggressive
behaviour against humans may raise the issue of human-
macaque conflict. Further studies on the specific impact
of crop raiding and feeding behaviour will derive the
implication of its conservation and management
strategies.
ACKNOWLEDGMENTS
The authors are grateful to the Ministry of
Environment and Forests, Government of India. The
logistic support provided by Divisional Forest officer,
Nicobar Division, Campbell Bay is duly acknowledged.
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1347 Journal of Research in Biology (2014) 4(4): 1338-1347
Article Citation: Duan HY, Wang CF, Yu YA, Li XW and Zhou YQ. Analysis on protein fingerprint, RAPD and fruit quality of tomato mutants by ion beam implantation. Journal of Research in Biology (2014) 4(4): 1348-1356
Jou
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al of R
esearch
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Biology
Analysis on protein fingerprint, RAPD and fruit quality of
tomato mutants by ion beam implantation
Keywords: Ion beam, tomato, SDS-PAGE, RAPD, fruit quality.
ABSTRACT: In this research, seeds of tomato were irradiated by ion beam or treated with ion beam and soybean DNA, and some tomato mutants with morphological variations were analyzed. Protein analysis in the leaves of mutants showed, changes of protein pattern in mutants were different as compared with the control, the main variation of protein pattern were darkening of bands, increase of protein bands were detected in mutant 12, mutant 14 and mutant 15 and lose of a band in mutant 15. Genomic DNA of mutants were analyzed by RAPD, and total number of amplification bands, number of differential bands and rate of differential bands were studied among mutants. Compared with the control, rate of differential bands was 100.0 % in mutant 9 and 15, also high in mutant 14 and 12, but was 20.0-50.0 % in other mutants except for mutant 3 and 11 without differential bands. In addition, content of vitamin C, soluble saccharide and protein were different, and fruit quality was multifarious in the fruit of mutants compared with the control; mutant 7 has better comprehensive nutritional quality of fruit, whereas mutant 12 and 14 stand second. The above results showed that effects of ion beam or soybean DNA on tomato genomic DNA would lead to the changes in gene expression, protein synthesis and fruit quality, moreover some tomato plants with better fruit quality or special characters were achieved, which would provide basis for the application of ion beam technology in tomato breeding.
1348-1356 | JRB | 2014 | Vol 4 | No 4
This article is governed by the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.0), which gives permission for unrestricted use, non-commercial, distribution and reproduction in all medium, provided the original work is properly cited.
www.jresearchbiology.com
Journal of Research in Biology
An International
Scientific Research Journal
Authors:
Duan HY*, Wang CF,
Yu YA, Li XW and
Zhou YQ.
Institution:
College of Life Science,
Henan Normal University,
Xinxiang 453007, China.
Corresponding author:
Duan HY.
Email Id:
Web Address: http://jresearchbiology.com/
documents/RA0454.pdf. Dates: Received: 03 Jun 2014 Accepted: 06 Jun 2014 Published: 26 June 2014
Journal of Research in Biology
An International Scientific Research Journal
Original Research
ISSN No: Print: 2231 –6280; Online: 2231- 6299
INTRODUCTION
In recent years, mutation breeding has been a
novel way in plant genetic improvement, especially low
energy ion beam implantation which exhibits many
advantages, such as low damage, high mutation rate,
wide mutational spectrum, and so on (Yu, 2000). At
present, ion beam mutation breeding technology has
been successfully applied to a lot of crop breeding, such
as rice, wheat, tobacco, cotton, soybean, rape and others
(Zhou, 2009). In addition, the etching and sputtering
effects of ion beam on cells would be very beneficial to
foreign DNA entering into the cells (Wang et al., 2009,
Li and Sun, 2011) and some transgenic plants have been
achieved by ion beam implantation (Duan et al., 2012),
thus the transgenic technology mediated by ion beam is a
simple and feasible transgenic method.
Tomato is one of the most important vegetables
and fruits that contain abundant nutrients, such as
lycopene, vitamin C, trace elements and other nutrients
(Xue et al., 2004, Wang et al., 2010). In order to meet
the need of people, germplasm resources or genetic
improvement breeding of tomato is required to be
studied and new cultivar of tomato should be cultivated.
In our laboratory, it was found that nitrogen ion (N+) or
argon ion (Ar+) had obvious influences on cell mitosis
and chromosome structure, and lead to various types of
chromosome aberrations (Duan et al., 2013). Thus, dry
seeds of tomato (tomato Zhongza No. 9) were irradiated
by N+ or Ar+ ion beam and soak into soybean DNA after
ion beam implantation to obtain a series of new
germplasm and cultivars with important application
value, and some tomato mutants with the variations of
morphologic characters were found in M1 present
generation. In this research, tomato mutants with
morphologic variations were analyzed by SDS-PAGE
and RAPD, and several indexes of fruit quality were also
detected, which would provide foundation for new
cultivars of tomato and theoretical basis for ion beam
mutation breeding of tomato.
MATERIALS AND METHODS
Plant materials
In this study, seeds of tomato (tomato Zhongza
No. 9) were provided by Vegetable Flower Institute of
Agricultural Sciences, Beijing, P. R. China, and were
respectively irradiated by N+ or Ar+ ion beam in the 30
kev energy conditions. Seeds of soybean (soybean
Zaoshu No. 2) were preserved in our laboratory, soybean
seedlings with single-leaf were used to extract genomic
DNA with CTAB method, and DNA fragments of
soybean genomic DNA were obtained by ultrasonication.
Culture of tomato plants
Tomato seeds implanted with N+ or Ar+ ion beam
were treated as described in research (Ji et al., 2001), at
first were respectively immersed into 0.1×SSC buffer
solution or 300 µg ml-1 DNA working solution which
was composed of soybean DNA and 0.1×SSC buffer
solution, and then were separately washed several times
with sterile water, but the control was only immersed
into sterile water. The above seeds were sowed in the test
field and cultured under the greenhouse conditions with
20°C light and 10°C dark temperature cycle. Seven days
later, seeds germinated, seedlings with two leaves were
transplanted in nutritive bowl and continued to be
cultured. When cultured for two months, seedlings with
five or six leaves were transplanted in the test field and
cultured at the above culture condition.
In addition, the variations of morphologic
characters in tomato plant were found, such as tall plant,
fat leaves, thick stalk, and so on, moreover protein and
DNA fingerprint of some tomato mutants were
respectively analyzed by SDS-PAGE or RAPD, and
several indexes of fruit quality were also detected.
SDS-PAGE of protein in leaves
Proteins were extracted from the fresh leaves of
tomato plants with morphologic variations as described
previously (Ji et al., 2001) with modifications. 1.0 g
leaves were mixed together with 1ml sterile water and
grinded in the mortar on ice-bath, and then the
Duan et al., 2014
1349 Journal of Research in Biology (2014) 4(4): 1348-1356
homogenate of leaves were centrifuged for 20 min by
12000 rpm at 4°C. The supernatant in the centrifuge tube
was transferred to 5 ml volumetric flask, furthermore, the
precipitate in the centrifuge tube was extracted again
with sterile water and then the supernatant was also
transferred to the above 5 ml volumetric flask, in which
the supernatant was diluted with sterile water to a
constant volume, then the solution was mixed and
preserved at -20°C. The content of soluble protein in the
above solution was determined by Bradford colorimetric
method (Bradford, 1976) at 595 nm, and the standard
curve of soluble protein was drawn with Bovine Serum
Albumin (BSA). In this research, SDS-PAGE of protein
was performed under experiment conditions of 3 %
stacking gel (pH6.8), 12 % separating gel (pH8.8) and
Tris-Glycine buffer solution (pH8.3), and Coomassie
Brilliant Blue method was used in this research.
RAPD amplification
In this study, leaves of tomato mutants were used
to extract DNA by CTAB extraction procedure (Ausubel
et al., 1987). RAPD amplification was performed as the
method (Kangfu et al., 1994). Reaction system of RAPD
amplification was 25 μl and composed of 20 ng DNA,
0.2 μmol L-1 primer, 0.2 μmol/L dNTPs, 2.0 mmol L-1
Mg2+, 1U Taq DNA polymerase and double distilled
water. RAPD amplification was performed as follows:
initial denaturalization at 94°C for 5 min, followed by 35
cycles of 94°C for 1 min, 36°C for 1 min and 72°C for
1.5 min, with a final extension cycle of 72°C for 8 min.
In addition, 100 primers were screened to obtain primers
by which amplification bands are most distinctive,
numbers of amplification bands are more and the
repeatability is preferable.
Determination of soluble saccharide in fruit
Assay of soluble saccharide was performed by
enthrone colorimetric method (Liu et al., 2013) with
improvement. Mature fruit of tomato mutants was
crushed with juicer, 0.5 g tomato juices were mixed
together with 5 ml sterile water in test tube, subsequently
the test tube was sealed with plastic film and put in
boiling water for 30 min to extract soluble saccharide.
The crude extract was filtered into 10 ml volumetric
flask, simultaneously the text tube and residues were
rinsed repeatedly with sterile water, and then the extract
was diluted with sterile water to constant volume. The
content of soluble saccharide was determined with
spectrophotometry at 485 nm, and the standard curve of
soluble saccharide was drawn with sucrose. In addition,
determination of soluble saccharide content was repeated
three times.
Determination of vitamin C and protein in fruit
Mature fruits of tomato mutants were crushed
with juicer, 0.5 g tomato juices were diluted with sterile
water to 100 ml volumetric flask, then extracted by
vacuum extrusion machine and preserved for the
determination of fruit protein and vitamin C.
Determination of fruit protein was performed as
determination of leaf protein in tomato, content of
vitamin C was assayed by spectrophotometry (Chen
et al., 2012) with modification and the standard curve of
vitamin C was drawn with standard vitamin C.
Moreover, determination of vitamin C and protein was
repeated three times.
RESULTS AND DISCUSSION
Protein fingerprint in the leaves of tomato mutants
It is well known that, effects of ion beam on
plant are very obvious and could cause versatility, such
as stem diameter, flowering phase, plant height, quality
characteristic, and so on (Phanchaisri et al., 2012). In this
research, protein in the leaves of tomato mutants were
analyzed by SDS-PAGE (Figure 1), and the electro
photograph was drawn in Figure 2 to more clearly
observe changes of the protein pattern. As compared
with the control, the main variation of protein pattern in
the mutants were some bands darkening, especially the
band with 0.350 Rf value obviously darkened, however
lose and increase of protein band was less found, only
Journal of Research in Biology (2014) 4(4): 1348-1356 1350
Duan et al., 2014
two bands increased in mutant 12, mutant 14 and mutant
15, and the Rf values were 0.05 and 0.083 respectively,
furthermore mutant 15 lost one band (Rf=0.133) in
comparison with the control and other mutants. The
above results suggest the effects of ion beam or soybean
DNA on leaf protein of tomato mutants were various,
which was same to other researchers (Ji et al., 2001).
Owing to the effects of ion beam on chromosome
structure (Huang et al., 1994), we infer that variation of
protein pattern in the leaves of tomato mutants might be
caused by the changes of genomic DNA due to the
damage of ion beam or integration of soybean DNA.
RAPD analysis of genomic DNA in tomato mutants
RAPD technology is actually PCR amplification,
and any organism could be identified by RAPD markers
(Williams et al., 1990, Welsh et al., 1991). Hither to,
some plant mutants induced by ion beam implantation
have been already analyzed by RAPD markers, such as
Nicotiana tabacum (Zhang et al., 1998), Cucumis melo
(Chen et al., 2002), Arabidopsis thaliana (Chang et al.,
2003), Dahlia pinnata Cav. (Yu et al., 2008), Jatropha
curcas (Pamidimarri et al., 2010), Balsamine (Gao et al.,
2012), and so on. In this research, genomic DNA of
tomato mutants was also analyzed with RAPD markers
in order to explore changes in the genomic DNA.
100 random primers were used in the RAPD
amplification, but only bands amplified by S11 primer
(GTAGACCCGT) and S45 primer (TGAGCGGACA)
could be variant between the control and tomato mutants,
and numbers of amplification bands and length of
amplification fragment were different in the mutants by
different primer (Figure 3). As shown in the Figure 3 (a),
only one DNA fragment with 550 bp was amplified by
primer S11 in the control, mutant 3 and mutant 11.
Compared with the control, DNA fragment with 850 bp
increased in mutant 2, mutant 4-8, mutant 10 and mutant
13, DNA fragment with 550 bp lost in mutant 9, mutant
12, mutant 14 and mutant 15, and numbers of
amplification bands and length of amplification fragment
were same in mutant 12 and mutant 14. Furthermore,
four DNA fragments were amplified in mutant 9, in
which DNA fragment with 700 bp was also amplified in
Duan et al., 2014
1351 Journal of Research in Biology (2014) 4(4): 1348-1356
M: marker, 1: the control, 2-11: tomato mutant induced by ion beam and soybean
DNA, 12: tomato mutant induced with 2×1017N+/cm2 ion beam, 13: tomato mutant
induced with 4×1017N+/cm2 ion beam, 14: tomato mutant induced with 2×1017Ar+/cm2
ion beam, 15: tomato mutant induced with 4×1017Ar+/cm2 ion beam.
Figure 1: Protein pattern in the leaves of tomato mutants by SDS-PAGE
mutant 12 and mutant 14. On the other side, bands
amplified by S45 primer were shown in Figure 3 (b); two
bands were amplified from the control, mutant 2-6,
mutant 11 and mutant 13, one special band was
amplified in mutant 8, mutant 10, mutant 12 and mutant
15 compared with the control. Moreover, three bands
were amplified in mutant 9, but their lengths were
different from the control and other mutants. Meanwhile,
there were two bands in mutant 14 in which one DNA
fragment with 700 bp was also found in mutant 12 and
mutant 15, yet other DNA fragment with 500 bp was
only amplified in mutant 14.
In addition, RAPD amplification bands of tomato
mutants by S11 and S45 primer were given in Table 1,
total number of amplification bands, number of
differential bands and rate of differential bands in tomato
mutants were found to be different. Compared with the
control, rate of differential bands were 100.0 % in mutant
9 and mutant 15, and number of differential bands were 7
and 3, respectively. Secondly, rate of differential bands
in mutant 14 and mutant 12 were also high, the number
of differential bands were 5 and 4, respectively.
However, rate of differential bands in the mutant 3 and
mutant 11 was 0.0 %, moreover rate of differential bands
in other mutants was in the scope of 20.0-50.0 %. Further
more, although rate of differential amplification bands
was 100.0 % in mutant 9, some protein bands only
darken and number of protein bands did not change in
mutant 9. In addition, the variation of protein pattern in
mutant 12, mutant 14 and mutant 15 were relatively
large, and rate of differential amplification bands was
respectively 66.7 %, 83.3 % or 100.0 %. Therefore, the
differential DNA fragments amplified by RAPD might
be related to the expression of some genes by encoding
some proteins or regulating protein synthesis, but it is not
clear whether differential DNA fragments could
influence fruit quality.
Fruit quality of tomato mutants
As everyone knows, tomato is rich in nutrition,
such as saccharide, vitamin C, protein, etc. (Xue et al.,
2004, Wang et al., 2010). In this research, fruit quality of
tomato mutants were assayed, content of vitamin C,
soluble saccharide and protein in the fruit of tomato
mutants were respectively listed in Table 2. As compared
Duan et al., 2014
Journal of Research in Biology (2014) 4(4): 1348-1356 1352
Figure 2: Protein ideograph in the leaves of tomato mutants
1: the control, 2-11: tomato mutant induced by ion beam and soybean DNA, 12: tomato mutant induced with
2×1017N+/cm2 ion beam, 13: tomato mutant induced with 4×1017N+/cm2 ion beam, 14: tomato mutant induced
with 2×1017Ar+/cm2 ion beam, 15: tomato mutant induced with 4×1017Ar+/cm2 ion beam.
with the control, content of vitamin C in 50 % mutants
was low, such as mutant 2-4, mutant 6, mutant 9, mutant
13 and mutant 15, especially lower in mutant 2, mutant 9
and mutant 4, and was 66.60 μg g-1, 69.65 μg g-1 and
74.43 μg g-1, respectively. However, content of vitamin
C was high in mutant 5, mutant 7, mutant 8, mutant 10-
12 and mutant 14, especially was higher in mutant 8
(152.03 μg g-1), mutant 10 (167.09 μg g-1) and mutant 12
(174.49 μg g-1), moreover content of vitamin C was the
highest in mutant 11 (242.24 μg g-1). In addition, content
of soluble saccharide in 64 % mutants was lower than the
control, but was high in mutant 2, mutant 5, mutant 7,
mutant 9 and mutant 10, particularly higher in mutant 7
(58.84 mg g-1) and mutant 2 (46.96 mg g-1). Furthermore,
content of protein was high in 64 % mutants in
comparison with the control, especially was the highest
Duan et al., 2014
1353 Journal of Research in Biology (2014) 4(4): 1348-1356
Figure 3: Results of RAPD amplification by S11 primer and S45 primer
(a) Results of RAPD amplification by S11 primer, (b) Results of RAPD amplification by S45 primer. M: DM2000,
M: marker, 1: the control, 2-11: tomato mutant induced by ion beam and soybean DNA, 12: tomato mutant induced
with 2×1017N+/cm2 ion beam, 13: tomato mutant induced with 4×1017N+/cm2 ion beam, 14: tomato mutant induced
with 2×1017Ar+/cm2 ion beam, 15: tomato mutant induced with 4×1017Ar+/cm2 ion beam.
a b
Table 1: RAPD amplification bands of tomato mutants by
S11 and S45 primer
Plants Total number
of bands
Number of
differential bands
Rate of differential
bands (%)
1 3 0 0.0
2 4 1 25.0
3 3 0 0.0
4 4 1 25.0
5 5 2 40.0
6 5 2 40.0
7 6 3 50.0
8 4 2 50.0
9 7 7 100.0
10 5 2 40.0
11 3 0 0.0
12 6 4 66.7
13 5 1 20.0
14 6 5 83.3
15 3 3 100.0
1: the control, 2-11: tomato mutant induced by ion beam and soybean
DNA, 12: tomato mutant induced with 2×1017N+/cm2 ion beam, 13: tomato
mutant induced with 4×1017N+/cm2 ion beam, 14: tomato mutant induced
with 2×1017Ar+/cm2 ion beam, 15: tomato mutant induced with 4×1017Ar+/
cm2 ion beam.
in mutant 9 (46.57 mg g-1), yet content of protein in
mutant 2, mutant 5, mutant 8, mutant 10 and mutant 11
was lower than the control, and only 6.17 mg g-1 protein
in mutant 10.
On the other side, content of vitamin C, soluble
saccharide and protein were different in mutants, and
fruit quality of mutants was multifarious. As shown in
Table 2, compared with the control, content of vitamin
C, soluble saccharide and protein in mutant 7 was all
higher, so mutant 7 has better comprehensive quality of
fruit, secondly were mutant 12 and mutant 14 because
content of vitamin C and protein was both higher.
Moreover, content of soluble saccharide and protein in
mutant 9 was both higher, especially content of protein
was the highest (46.57 mg g-1). However content of
vitamin C in mutant 11 was the highest (242.24 μg g-1),
and content of soluble saccharide and protein was only
11.06 mg g-1 and 18.58 mg g-1. In addition, content of
vitamin C and soluble saccharide was low in mutant 15
and mutant 3, one other thing to note is that nutritional
quality of mutant 3 and mutant 11 are obviously different
with the control, but rate of differential amplification
bands was 0.0 % in mutant 3 and 11 which were treated
with ion beam and soybean DNA, inferring some big
insert segment of soybean DNA might be not amplified,
perhaps there might be a more complicated relationship
between nutritional quality of fruit and genomic DNA of
tomato irradiated with ion beam or treated with ion beam
and soybean DNA, moreover the effect mechanism of
ion beam or foreign DNA was very complex and need to
be further studied and explored.
CONCLUSION
This study shows that ion beam or soybean DNA
could influence leaf protein, genomic DNA and fruit
quality of tomato mutants, inferring the variation of leaf
protein and fruit quality in tomato mutants might be
caused by the changes of genomic DNA which would
happen due to damage of ion beam or integration of
soybean DNA. However the effects of ion beam or
Journal of Research in Biology (2014) 4(4): 1348-1356 1354
Duan et al., 2014
Table 2: Content of vitamin C, soluble saccharide and protein in the fruit of tomato
Plant Content of vitamin C
(μg/g)
Content of soluble
saccharide (mg/g)
Content of protein
(mg/g)
1 111.95 19.18 18.88
2 66.60 46.96 13.98
3 95.07 13.17 20.51
4 74.43 17.09 26.44
5 114.28 21.37 18.48
6 95.66 14.19 20.12
7 116.91 58.84 29.19
8 152.03 16.35 17.45
9 69.65 37.48 46.57
10 167.09 40.51 6.17
11 242.24 11.06 18.58
12 174.49 16.46 25.48
13 92.36 13.65 21.19
14 122.95 19.12 24.86
15 99.96 12.62 20.35
The average content
in mutants 119.71 23.87 21.88
1: the control, 2-11: tomato mutant induced by ion beam and soybean DNA, 12: tomato
mutant induced with 2×1017N+/cm2 ion beam, 13: tomato mutant induced with 4×1017N+/cm2
ion beam, 14: tomato mutant induced with 2×1017Ar+/cm2 ion beam, 15: tomato mutant
induced with 4×1017Ar+/cm2 ion beam.
soybean DNA were different, and the changes among
protein, DNA and fruit quality was not consistent with
each other, thus it is necessary to further study effect
mechanism of ion beam or foreign DNA, which would
contribute to provide foundation for ion beam mutation
breeding of tomato.
ACKNOWLEDGMENT
This research was kindly supported by Science
Fund from Henan province (122300410025), and the
grant of young teachers in Henan province institution of
higher learning (2011GGJS-063), in P. R. China.
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Article Citation: Marina de Sá Leitão Câmara de Araújo. The leaping behavior of the sally lightfoot crab Grapsus grapsus (Crustacea: Decapoda: Brachyura) at an oceanic archipelago. Journal of Research in Biology (2014) 4(4): 1357-1364
Jou
rn
al of R
esearch
in
Biology
The leaping behavior of the sally lightfoot crab Grapsus grapsus
(Crustacea: Decapoda: Brachyura) at an oceanic archipelago
Keywords: Crab behavior, Fernando de Noronha Archipelago, Red rock crab, Semi-terrestrial crab.
ABSTRACT: The genus Grapsus includes a total of nine recognized species of semi-terrestrial crabs. Among them, Grapsus grapsus (Linnaeus, 1758) stands popularly known as sally lightfoot crab. It is very abundant in Oceanic Islands, such as the Fernando de Noronha Archipelago, Brazil. The present study registered the behavior of jumping between the rocks by G. grapsus in the supralittoral of Fernando de Noronha Archipelago. Field observations were performed in May 2012, including video footage. The crabs, juveniles and adults, males and females, jump from a rock to another. This can be related to a defense habit, but it seems that the crabs also jump to avoid entering into the sea, or to escape from wave wash. Other registers on crabs jumping from literature are also discussed. However, more studies on this behavior are still necessary for understanding them completely.
1357-1364 | JRB | 2014 | Vol 4 | No 4
This article is governed by the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.0), which gives permission for unrestricted use, non-commercial, distribution and reproduction in all medium, provided the original work is properly cited.
www.jresearchbiology.com Journal of Research in Biology
An International
Scientific Research Journal
Authors:
Marina de Sá Leitão
Câmara de Araújo.
Institution:
Departamento de Ciências
Exatas e Naturais, Faculdade
de Ciência, Educação e
Tecnologia de Garanhuns
(FACETEG), Campus
Garanhuns, Universidade de
Pernambuco (UPE), Brazil.
Corresponding author:
Marina de Sá Leitão
Câmara de Araújo.
Email Id:
Web Address: http://jresearchbiology.com/
documents/RA0452.pdf.
Dates: Received: 20 May 2014 Accepted: 30 May 2014 Published: 26 Jun 2014
Journal of Research in Biology
An International Scientific Research Journal
Original Research
ISSN No Print: 2231 –6280; Online: 2231- 6299
INTRODUCTION
The genus Grapsus Lamarck, 1801 (Grapsidae)
includes a total of nine recognized species of semi-
terrestrial crabs: G. adscensionis (Osbeck, 1765),
G. albolineatus Latreille, in Milbert, 1812,
G. fourmanoiri Crosnier, 1965, G. granulosus H. Milne
Edwards, 1853, G. grapsus (Linnaeus, 1758),
G. huzardi Desmarest, 1825, G. intermedius de Man,
1888 , G. longi tars is Dana , 1851 and
G. tenuicrustatus (Herbst, 1783) (WORMS, 2013; Ng
et al., 2008). Among these species, G. grapsus, stands
out popularly and are known as red rock crab, sally
lightfoot crab, "aratu" (in Portuguese) and "abuete negro"
or "sayapa" (in Spanish). This species is found in the
Pacific Ocean, from Baja California to Northern Chile,
and Galapagos Islands, and in the Atlantic Ocean, from
Bermudas, Florida, Gulf of Mexico, Antilles, Colombia,
and from Venezuela to Brazil. In the Brazilian coast, this
crab is found from the States of Ceará to Espírito Santo,
but it is more abundant in the Oceanic islands (Fernando
de Noronha Archipelago, Rocas Atoll and Saint Peter
and Saint Paul Rocks) (Melo, 1996; Freire et al., 2011).
At Saint Peter and Saint Paul Rocks, (Ross 1847, apud
Holthuis et al., 1980) cited that this species is a predator
of the eggs of birds that nest at the area, and Viana et al.,
(2004) cited that this is one of the most abundant animal
species on the rocks. Melo (1996) also signals the
occurrence of this species at Trindade, a Brazilian
volcanic island distant 1,167 km from the continent, but
probably the species inhabiting this island is, in fact,
G. adscensionis (Hartnoll, 2009). Ratti (2004) believed
that the differences between G. adscensionis and
G. grapsus were not enough to support two different
species, but more recently, several authors such as Ng
et al., (2008) and Freire et al., (2011), recognized the
taxonomic validity of both species.
Among the oceanic island this species can be
found, stands out the Fernando de Noronha Archipelago
(FNA) (3°51′S, 32°25′ W), a complex of volcanic islands
and rocks, which is found under jurisdiction of the State
of Pernambuco, Northeast of Brazil. The benthic fauna
of FNA was studied by Lopes and Alvarenga (1955) and
Matthews and Kempf (1970) (Mollusca), Pires et al.,
(1992) (Cnidaria), Mothes and Bastian (1993) and
Muricy and Moraes (1998) (Porifera), among others.
Several oceanographic expeditions explored the
archipelago, such as H.M.S. Beagle Challenger
Expedition, Hartt Expedition, Branner-Agassiz
Expedition, Calypso, Canopus and Almirante Saldanha.
The results of the Crustacea sampled on these
expeditions can be found at several publications, such as
Smith (1869), Miers (1886), Henderson (1888), Bate
(1888), Rathbun (1900, 1918, 1925, among others),
Forest and de Saint-Laurent (1967) and Coelho et al.,
(2006, 2007, 2008). Fausto-Filho (1974) presented a list
of the Decapoda and Stomatopoda collected by himself
and based on some of the cited publications, which
resulted in a total of 66 species (3 Stomatopoda and 63
Decapoda) for FNA. Included, there is G. grapsus. The
species was considered very abundant, being found in all
beaches. There is no doubt that the species inhabiting
FNA is G. grapsus. They are commonly observed in the
rocky shores of the islands that compose the archipelago,
sharing the habitat with Plagusia depressa (Fabricius,
1775) (Plagusiidae). The present study aims to describe
the jumping behavior of Grapsus grapsus at FNA during
field observations.
MATERIAL AND METHODS
The archipelago is distant 545 km from the
capital of Pernambuco, the Municipality of Recife,
occupies an area of 26 km² and the main island,
Fernando de Noronha, has an area of 17 km², being 6
miles long and 2 miles wide (Matthews and Kempf,
1970; Fausto-Filho, 1974). In May 2012, during three
days, field observations and footages of this species were
performed at Sueste Bay, FNA (Figure-1) (3º52'01" S;
32º25'19" W). At the bay, the Sueste Beach and the
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1358 Journal of Research in Biology (2014) 4(4): 1357-1364
Sueste Mangrove are included, the last one being
considered the only oceanic mangrove of South Atlantic.
In the seawater of the bay, there are several islets, such
as Cabeluda, Chapéu, Ovos and Trinta-Réis.
The individuals of Grapsus grapsus were
observed in the rocky shore of the bay. These rocks are
mainly distributed in the extremities of the bay, and also
serve as habitat for Plagusia depressa. The water was
transparent and shallow, with a depth of 1m. The footage
was performed with a Panasonic camera, DMC-FT10
model. After that, a bibliographic research was
performed to seek possible registers of the jumping
behavior of crabs in the literature.
The air temperature and tidal heights for the
dates of study were obtained through the Integrated
System of Environmental Data (SINDA).
RESULTS AND DISCUSSION
The air temperature for the study period varied
from 25.5 to 30ºC (Figure-2), characterizing a tropical
climate. The observations were performed during the dry
period, equatorial summer. According to Ribeiro et al.,
(2003, 2005), the FNA climate is of the type Aw of
Köppen's classifications, i.e. tropical with semi-arid
Journal of Research in Biology (2014) 4(4): 1357-1364 1359
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A B
C D
Figure 1. Brazilian coast with the location of the Fernando de Noronha Archipelago, FNA
(A); FNA with the location of the Sueste Bay (B); Aerial view of the Sueste Bay (C);
Rocky shore at Sueste Bay, where the field observations of Grapsus grapsus (Linnaeus,
1758) were perfomed (D).
characteristics, having well defined dry and rainy
periods.
The tidal level for the study period varied from
1.25 to 2.75 m (Figure-3). The tidal regime can be
characterized as semi-diurnal tide, since there are two
high tides in each lunar day (Thurman, 1997). According
to Souza (2011), the maximum height of the tide in FNA
is 2.80 m, and the minimum, 0.0m. Thus, regarding its
amplitude, the tide of FNA can be classified as
mesotides.
The observed population consisted of Grapsus
grapsus juveniles and adults of both sexes. They were
found sharing the habitat with Plagusia depressa.
Besides the size, adults and juveniles are also
distinguished by the color of the carapace. Juveniles of
G. grapsus are dark green, dark gray or almost black,
which is important for they camouflage on the
black volcanic rocks of oceanic islands, and with light
yellow spots. On the other hand, adults are quite variable
in color; some are dark red or bright red (especially
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1360 Journal of Research in Biology (2014) 4(4): 1357-1364
Figure 2. Air temperature by dates and hour during the study period,
at Fernando de Noronha Archipelago.
Figure 3. Tidal level by dates and hour during the study period,
at Fernando de Noronha Archipelago.
males), others are dark green. Some lines and spots can
be observed (Fausto-Filho, 1974; Freire et al., 2011)
(Figure-4).
During the field observations, an unusual
behavior in Brachyura could be noticed: the sally
lightfoot jumps from a rock to another. Two scenes of
G. grapsus jumping were recorded (Videos 1, 2 and 3).
This behavior was observed for both males and
females, and juveniles and adults. A total of 12
observations were performed. In a first moment, it can be
an useful strategy to prevent predation, as described to
the species which will be discussed below. Besides, this
type of movement could be important to escape from the
wave wash (Video 1) or to avoid entering into the water
(Video 2), instead of walking through the water to reach
another point of the rocks. They also seem to jump from
a lower to a higher rock (Video 3). Kramer (1967) also
observed this behavior in a population of G. grapsus
from Galapagos. He noticed that the jumpy crabs had an
average carapace width of 30 cm. The crabs from FNA
were not measured, but it was clear that they did not
reach 10 cm CW. Before jumping, the crab aligns the
body by stretching the front running pairs of legs on
(Kramer, 1967), which was also noticed in the present
study.
Some other interesting information was found in
the literature, regarding the locomotion of crabs. The
species Armases roberti (H. Milne Edwards, 1853)
(Sesarmidae) is found along river banks between rocks
and stones, as well as on the vegetation (Chace and
Hobbs, 1969). According to Schubart and Diesel (1998),
when these crabs are disturbed, they jump from the trees
into the water, and due to this behavior, they are know in
the Caribbean as “jumpy crabs”. Thus, this behavior
could be related to a defensive attitude. A similar
behavior was also registered for Percnon gibbesi (H.
Milne-Edwards, 1853) (Percnidae) by Deudero et al.,
(2005); the specimens, observed in shallow waters, run
and jump when threatened, seeking for shelter from
predators.
The crabs Sesarma trapezoideum H. Milne
Edwards, 1837 (Sesarmidae) occur preferentially in
riverine cliffs near water streams (Jeng et al., 2003).
According to these authors, these crabs retreat into
crevices or jump into the water below them when
disturbed; few minutes after that, they climb back to the
cliff. The species Leptograpsus variegatus (Fabricius,
1793) (Grapsidae), a supralittoral crab of rocky shores as
G. grapsus, jump into tidal pools or into the sea to escape
from predation (Greenaway et al., 1992).
CONCLUSIONS
All these mechanisms described in literature are
related to a fast escape from danger, such as predation,
including jumping into the water. But during the field
observations of G. grapsus, it could be noticed that the
specimens also jump from a rock to another, which could
be useful to escape from the wave wash or to avoid
entering into the water. They also seem to jump to a
higher rock. However, further studies on this feature are
still necessary. For example, to test if there is difference
in the jumping frequency between sexes and age classes,
as well as or to correlate the distance or amplitude of the
jumps with the body size of the crab.
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Journal of Research in Biology (2014) 4(4): 1357-1364 1361
Figure 4. Crabs of the species Grapsus grapsus
(Linnaeus, 1758) from the rocky shore at Sueste
Bay, Fernando de Noronha Archipelago.
ACKNOWLEDGEMENT
The author is thankful to Maurício de Sá Leitão
Dévé, Silvia de Sá Leitão Dévé e Jean Luc Dévé for
aiding in the field work and footage of the species. I also
thank Dr. Christoph Schubart for bringing me
informations regarding crabs' behavior, which helped me
describing the 'jumpy' grapsoids of Fernando de Noronha
Archipelago.
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