alexa Inter-Specific Variation Studies among Nephrolepis using SDS-PAGE | OMICS International
ISSN: 2472-0992
Journal of Pharmacognosy & Natural Products

Like us on:

Make the best use of Scientific Research and information from our 700+ peer reviewed, Open Access Journals that operates with the help of 50,000+ Editorial Board Members and esteemed reviewers and 1000+ Scientific associations in Medical, Clinical, Pharmaceutical, Engineering, Technology and Management Fields.
Meet Inspiring Speakers and Experts at our 3000+ Global Conferenceseries Events with over 600+ Conferences, 1200+ Symposiums and 1200+ Workshops on
Medical, Pharma, Engineering, Science, Technology and Business

Inter-Specific Variation Studies among Nephrolepis using SDS-PAGE

Johnson M*

Centre for Plant Biotechnology, Department of Botany, St. Xavier’s College (Autonomous), Palayamkottai, Tamil Nadu, India

*Corresponding Author:
Marimuthu Johnson
Centre for Plant Biotechnology, Department of Botany
St. Xavier’s College (Autonomous)
Palayamkottai, Tamil Nadu, India
Tel: +979786924334
Fax: +914622561765
E-mail: [email protected]

Received date: October 08, 2015; Accepted date: September 06, 2015; Published date: September 10, 2015

Citation: Johnson M (2015) Inter-Specific Variation Studies among Nephrolepis using SDS-PAGE. J Pharmacogn Nat Prod 2:112. doi: 10.4172/2472-0992.1000112

Copyright: © 2015 Johnson M. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.

Visit for more related articles at Journal of Pharmacognosy & Natural Products

Abstract

The present study was aimed to reveal the biochemical similarity and varaition among the three taxonomically confused species Nephrolepis exaltata (L.) Schott, Nephrolepis auriculata (L.) Trimen and Nephrolepis multiflora (Roxb.) Jarret using SDS-PAGE analysis. To reveal the inter-speciifc variation among the selected Nephrolepis, SDS-PAGE was carried out to obtain protein bands following the method described by Anbalagan. Multiple regions (8) of activity were observed from protein  electrophoretic system of Nephrolepis. A total of 19 bands with various Rf values and molecular weight were demonstrated in the SDS-PAGE gel system of Nephrolepis. Among the three species of  Nephrolepis, N. exaltata showed maximum number of protein bands (9) followed by N. multiflora (6) and N. auriculata (4). Each region expressed different proteins which act as representative of the expression of a particular gene in the studied species of Nephrolepis. The similarity indices were calculated and cladogram was constructed based on the protein profiles of Nephrolepis and revealed the similarities and variation among the studied Nephrolepis species. The results obtained in this work also showed that SDS-PAGE analysis can provide an easy, low cost and quick way for the identification of inter-specific variation among the selected Nephrolepis species. SDS-PAGE analysis provided strong basis for the discrimination of genotypes on the basis of specific polypeptide fragments.

Keywords

Nephrolepis; Protein profile; SDS-PAGE

Introduction

Nephrolepis is a tropical genus with 40 species is identified worldwide (Friedrich 2005). In India nearly eight species are enumerated [1]. Among the eight speceis, except Nephrolepis delicatula all other species recorded in South India. But Manicakam and Irudayaraj [2] noted only two indigenous Nephrolepis species, other species nomenclature, identity and distribution has to be confirmed. Similarly Sledge [3] also pointed out the nomenclatural problem of Nephrolepis cordifolia, Nephrolepis auriculata and Polystichum auriculatum. Mickel and Beitel [4] also noted the misidentification of Nephrolepis exaltata and Nephrolepis biserrata. Nephrolepis multiflora (Roxb.) Jarret is distinguished morphologically by the charactereous frond and finely granulose exine. Nephrolepis auriculata (L.) Trimen is identified morphoogically based on the herbaceous fronds, exine with reticulate ridges and tubers on the roots. Nephrolepis exaltata (L.) Schott, with alternate pinnae (the small “leaflets” on either side of the midrib), each pinna being 2-8 cm long. The pinnae of N. exaltata, are generally deltoid and the edges appear slightly serrate. Similar to N. multiflora the N. exaltata also not possesed the tubers in the roots. Nephrolepis exaltata (L.) Schott, Nephrolepis auriculata (L.) Trimen and Nephrolepis multiflora (Roxb.) Jarret are morphologically similar and they posess taxonomic problems especially in their identification. Much more studies therefore, need to be carried out to provide taxonomic features that will delimit the species. Polyacrylamide Gel Electrophoresis (PAGE) is a versatile biochemical technique to detect genetic variation. In recent years, there has been explosion in the availability of different types of genetic markers [5]. With this knowledge the study was aimed to reveal the biochemical similarity and varaition among the three taxonomically confused species Nephrolepis exaltata (L.) Schott, Nephrolepis auriculata (L.) Trimen and Nephrolepis multiflora (Roxb.) Jarret using SDS-PAGE anaysis.

Materials and Methods

For the electrophoresis studies, 500 mg of Nephrolepis exaltata (L.) Schott, Nephrolepis auriculata (L.) Trimen and Nephrolepis multiflora (Roxb.) Jarret young individual croziers were harvested from Kodaikannal Botanic Garden, Eettipallum, Kodaikannal and ground on ice cold mortar and pestle with 0.1 M phosphate buffer (pH 7.0). The resultant slurry was centrifuged at 10,000 rpm for 10 min at 4ºC in cooling centrifuge and the supernatant was stored at -70ºC before use. SDS-PAGE was carried out to obtained protein bands following the method described by Anbalagan [6]. After electrophoresis the gel was observed using a Vilber Loubermat gel documentation system and banding profiles of protein was compared by zymogram. For the interspecific relationship studies, the protein profile was converted into a “1” and “0” matrix, to indicate the presence or absence of the Rf Values, respectively. Genetic similarities (GS) were estimated according to Nei and Li [7]. To demonstrate the inter-specific relationship among the studied Nephrolepis, a dendrogram was constructed by UPGMA using NTSYSpc-2.0 software.

Results and Discussion

The relative positions of the protein bands of the studied Nephrolepis species viz., Nephrolepis exaltata (L.) Schott, Nephrolepis auriculata (L.) Trimen and Nephrolepis multiflora (Roxb.) Jarret collected from various localities of South India were revealed by SDSPAGE. Multiple regions (8) of activity were observed from protein electrophoretic system of Nephrolepis. A total of 19 bands with various Rf values and molecular weight were demonstrated in the SDS-PAGE gel system of Nephrolepis (Table 1; Figure 1). Among the three species of Nephrolepis, N. exaltata showed maximum number of protein bands (9) followed by N. multiflora (6) and N. auriculata (4). The observed protein profile demonstrated the role of protein in similarity and variation between the studied species of Nephrolepis. Each region expressed different proteins which act as representative of the expression of a particular gene in the studied species of Nephrolepis. Based on the occurrence of proteins in the Nephrolepis gel system, the protein profiles were classified into ten regions. Region 1 observed with only one protein (PP11) with Rf- 0.02 and MW- 177.8 KDa) showed its presence in N. exaltata and N. multiflora. Region 1 explained the similarity between the N. exaltata and N. multiflora. Region 2 showed four protein bands with three positions (PP21-3). PP21 (Rf-0.10; MW-162.2 KDa) and PP22 (Rf-0.16; MW-123.0 KDa) was dispalyed their existence only in N. exaltata. PP23 (Rf-0.18; MW-100.0 kDa) represented its common occurrence in N. exaltata and N. multiflora. Two unique bands were found in region 2 and explained the uniquness of N. exaltata. Region 3 depicted four bands with four positions. PP31 (Rf-0.2; MW-99.5 KDa) expressed its unique occurrence in N. auriculata. Similarly PP32 (Rf-0.22; MW-96.2 KDa) was present only in N. multiflora. PP33 (Rf-0.24; MW- 93.3 KDa) and PP34 (Rf-0.29; MW- 87.0 KDa) was showed their existence only in N. exaltata. In region 3, the studied three species explained its exclusive character with the distinct Rf value and protein occurrence in the protein gel system of Nephrolepis. Region 4 represented four bands with three positions. PP41 (Rf-0.31; MW-80.3 KDa) was showed its occurrence only in N. auriculata. PP42 (Rf-0.35; MW-61.6 KDa) was demonstrated its unique existence only in N. exaltata. PP43 (Rf-0.37; MW-58.5 KDa) showed its common presence in N. auriculata and N. exaltata. N. multiflora failed to express its occurrence in this region. The region 4 clearly explained the biochemical similarities and difference between N. auriculata and N. exaltata. Region 5 depicted two bands with two positions only. PP51 (Rf-0.43; MW-50.1 KDa) was distinct to N. multiflora that explained its special occurrence in the region 5. Similarly PP52 (Rf-0.47; MW-46.1 KDa) was observed in N. exaltata. In this region N. auriculata failed to demonstrate its existence. The region 5 also clearly distinguished the difference among the studied species with specific protiens. Region 6 (PP61 - Rf-0.55; MW-39.2 KDa), 7 (PP71 - Rf-0.65; MW-29.6 KDa)) and 8 (PP81 - Rf-0.72; MW-25.1 KDa)) represented with only one proteins. The protein PP61 and PP71 showed their occurrence only in N. multiflora. PP81 (Rf-0.72; MW-25.1 KDa) displayed its unique presence in N. auriculata only which explained its variation with other studied species of Nephrolepis. Regions 9 and 10 were failed to express protein occurence in the studied Nephrolepis species. The banding patterns of proteins in the SDS-PAGE gel system of Nephrolepis discussed the similarity and variation between the studied three species of Nephrolepis. The similarity indices were calculated and cladogram was constructed based on the protein profiles of Nephrolepis and revealed the similarities between the N. multiflora and N. exaltata, the N. auriculata showed the variation and occupied a separate clade in the cladogram (Figure 2). The evolutionary tree which constructed based on the protein profile expressed two clusters (C1 and C2). The cluster (C1) includes two species of Nephrolepis viz., N. multiflora and N. exaltata. The cluster (C2) is simply with one species N. auriculata, the cladogram constructed based on the protein profile showed the closeness and divergence among the three Nephrolepis species. Among the various analytical tools, electrophoresis is a relatively simple, rapid and highly sensitive tool to study the properties of proteins and nucleic acids. Studies of protein variation are important tool (PAGE) that has often been employed to know the biochemical variation, inter and intra specific variation and evolutionary relationships among the plant species [8-13]. In addition, researchers employed the Poly Acrylamide Gel Electrophoresis (PAGE) to know information about the molecular weights and charges of proteins. Protein electrophoresis is a powerful tool for population genetics [14]. Ghafoor et al. [15] employed protein profiles as genetic markers to resolve taxonomic and evolutionary problems of Cicer arietinum. Harendra Singh et al. [16] differentiated Rhizobium inoculated Desi and Kabuli Chickpea (Cicer arietinum L.) genotypes using SDS-PAGE. Dudwadkar et al. [17] enumerated the protein diversity of Cucurbitaceae using SDS-PAGE protein profile and found the similarities and variations among the Cucurbitaceae members. Sivaraman et al. [18] and Revathy et al. [19] revealed the protein expression on various morphogenetic developments of selected ferns from Western Ghats. Narayani and Johnson [20] studied inter specific proteomic studies on selected Selaginella species using SDS-PAGE. In the present study also SDS-PAGE protein profiles distinguished the similarity and variation among the three Nephrolepis species. The results of the present study also supplemented the previous observation and application by expressing inter-specific protein variation among the three Nephrolepis species. These banding profiles can be used as biochemical and pharmacognostical marker to distinguish the medicinally important Nephrolepis from its adulterants in the pharmaceutical industries. Characterization of genetic diversity in Nephrolepis species, offers an opportunity for breeders for its exploitation in wide hybridization programs in the horticultre. Furthermore, the results obtained in this work also showed that SDS-PAGE analysis can provide an easy, low cost and quick way for the identification of inter-specific variation among the selected Nephrolepis species. SDS-PAGE analysis provided strong basis for the discrimination of genotypes on the basis of specific polypeptide fragments.

MW - Rf Mol. Wt. in K Da Region Position N. multiflora N. auriculata N. exaltata
0.08 177.8 1 PP11 +   +
0.10 162.2 2 PP21     +
0.16 123.0 PP22     +
0.18 100.0 PP23 +   +
0.20 99.5 3 PP31   +  
0.22 96.2 PP32 +    
0.24 93.3 PP33     +
0.29 87.0 PP34     +
0.31 80.3 4 PP41   +  
0.35 61.6 PP42     +
0.37 58.5 PP43   + +
0.43 50.1 5 PP51 +    
0.47 46.1 PP52     +
0.55 39.2 6 PP61 +    
0.65 29.6 7 PP71 +    
0.72 25.1 8 PP81   +  

Table 1: Protein profile of Nephrolepis species in SDS-PAGE gel system.

pharmacognosy-natural-products-SDS-PAGE-pattern

Figure 1: SDS PAGE pattern and zymogram of SDS-PAGE protein profile of Nephrolepis.

pharmacognosy-natural-products-protein-profile-Nephrolepis

Figure 2: Cladogram based on the protein profile of Nephrolepis.

References

Select your language of interest to view the total content in your interested language
Post your comment

Share This Article

Recommended Conferences

Article Usage

  • Total views: 12195
  • [From(publication date):
    February-2016 - Jul 23, 2018]
  • Breakdown by view type
  • HTML page views : 8362
  • PDF downloads : 3833
 

Post your comment

captcha   Reload  Can't read the image? click here to refresh

Peer Reviewed Journals
 
Make the best use of Scientific Research and information from our 700 + peer reviewed, Open Access Journals
International Conferences 2018-19
 
Meet Inspiring Speakers and Experts at our 3000+ Global Annual Meetings

Contact Us

Agri & Aquaculture Journals

Dr. Krish

[email protected]

+1-702-714-7001Extn: 9040

Biochemistry Journals

Datta A

[email protected]

1-702-714-7001Extn: 9037

Business & Management Journals

Ronald

[email protected]

1-702-714-7001Extn: 9042

Chemistry Journals

Gabriel Shaw

[email protected]

1-702-714-7001Extn: 9040

Clinical Journals

Datta A

[email protected]

1-702-714-7001Extn: 9037

Engineering Journals

James Franklin

[email protected]

1-702-714-7001Extn: 9042

Food & Nutrition Journals

Katie Wilson

[email protected]

1-702-714-7001Extn: 9042

General Science

Andrea Jason

[email protected]

1-702-714-7001Extn: 9043

Genetics & Molecular Biology Journals

Anna Melissa

[email protected]

1-702-714-7001Extn: 9006

Immunology & Microbiology Journals

David Gorantl

[email protected]

1-702-714-7001Extn: 9014

Materials Science Journals

Rachle Green

[email protected]

1-702-714-7001Extn: 9039

Nursing & Health Care Journals

Stephanie Skinner

[email protected]

1-702-714-7001Extn: 9039

Medical Journals

Nimmi Anna

[email protected]

1-702-714-7001Extn: 9038

Neuroscience & Psychology Journals

Nathan T

[email protected]

1-702-714-7001Extn: 9041

Pharmaceutical Sciences Journals

Ann Jose

[email protected]

1-702-714-7001Extn: 9007

Social & Political Science Journals

Steve Harry

[email protected]

1-702-714-7001Extn: 9042

 
© 2008- 2018 OMICS International - Open Access Publisher. Best viewed in Mozilla Firefox | Google Chrome | Above IE 7.0 version
Leave Your Message 24x7