Morphological variations in teak (Tectona grandis L. f.) clones

Views:
 
Category: Entertainment
     
 

Presentation Description

No description available.

Comments

Presentation Transcript

slide 1:

259 International Journal of Chemical Studies 2018 65: 259-264 P-ISSN: 2349 –8528 E-ISSN: 2321 –4902 IJCS 2018 65: 259-264 © 2018 IJCS Received: 21-07-2018 Accepted: 25-08-2018 Modi JS Department of Silviculture and Agroforestry ASPEE College of Horticulture and Forestry Navsari Agricultural University Navsari Gujarat India Tandel MB Department of Silviculture and Agroforestry ASPEE College of Horticulture and Forestry Navsari Agricultural University Navsari Gujarat India Prajapati VM Department of Silviculture and Agroforestry ASPEE College of Horticulture and Forestry Navsari Agricultural University Navsari Gujarat India Parekh VB Department of Silviculture and Agroforestry ASPEE College of Horticulture and Forestry Navsari Agricultural University Navsari Gujarat India Ahir BR Department of Silviculture and Agroforestry ASPEE College of Horticulture and Forestry Navsari Agricultural University Navsari Gujarat India Correspondence Modi JS Department of Silviculture and Agroforestry ASPEE College of Horticulture and Forestry Navsari Agricultural University Navsari Gujarat India Molecular variations in teak Tectona grandis L. f. clones Modi JS Tandel MB Prajapati VM Parekh VB and Ahir BR Abstract The investigation entitled “Molecular variations in Teak Tectona grandis L. f. clones” was carried out at Rajpipla Silviculture Forest Division Rajpipla and Biotechnology Laboratory ASPEE College of Horticulture and Forestry Navsari Agricultural University Navsari Gujarat. The experiment comprised of 15 clones viz. C1-Bandhpada North Dangs C2-Chikhli South Dangs C3-Nilambo Dungarda C4- African Dungarda C5-Satkhasi Vyara C6-Khalta Bariya C7-Achhala Godhra C8-Kevadi Chotaudepur C9-Raighadh Sabarkantha North C10-Danta Banaskantha C11-Vanaj South Sabarkantha C12-Dankiwadu Gir West C13-Bhavnath Junaghadh C14-Develvel Gir East and C15- Khatam Rajpipla East. This experiment was laid out in Randomized Block Design RBD comprising of fifteen clones and three replications. The seven decamer primers produced a total of 50 scorable bands in the fifteen clones of Tectona grandis L. f. out of them 20 were polymorphic and 30 were monomorphic. The percentage of polymorphism ranged from a maximum 50.00 by OPE-18 to a minimum of 16.67 by OPE-10. The lowest genetic similarity 0.7209 was between the clones 4 7 whereas the highest genetic similarity 0.9761 was between the clones 9 10. The dendrogram of fifteen clones reflected that the clones were divided in two main clusters named A and B. Further Cluster-B was divided in to two sub clusters B1 and B2. The clones C7 and C8 reported in Cluster-A which were found to be more diverse as compared to other clones. The clones C1 C3 C5 C2 C6 and C4 were observed in sub-cluster-B1. Whereas clones C9 C10 C15 C11 C12 C13 and C14 were recorded in sub-cluster-B2. The clones C4 and C14 found in B1 and B2 sub-cluster respectively which having minimum similarity with other clones. Keywords: molecular variations teak Tectona grandis L. f. and clones Introduction The forests of India are ancient in nature with high diversity. They are not only rich in tree species composition but also provide shelter to a wide range of fauna avian-fauna and insects. The teak tree is native to South-East Asia more specifically to India Myanmar Thailand and Laos. Over the past 150 years it has been planted extensively both within its native range and in other tropical and sub-tropical region of Asia Africa and America. It is naturalized in the Indonesian island of Java and some of the smaller islands east of Java where it is believed to have been introduced some 400-600 years ago. Teak Tectona grandis is a tall and handsome deciduous tree representing Lamiaceae family in plant kingdom. Locally it is also known as Sagon Saigon Saj Taku Kayum etc. in various Indian languages. The teak tree is well known for its versatile timber. Its heartwood combines several qualities like termite and decay resistance lightness and strength drying without warping and splitting easy workability and attractive appearance making it one of the world’s finest timbers. Teak genetic improvement was started in India in the year 1954. Teak improvement was limited to establishing seed production areas SPAs and clonal seed orchards CSO. CSOs are established with grafted plants of superior phenotypes plus trees selected from natural forests and plantations. These "plus trees" were chosen at a very high intensity often one in several hectares of forests with rigorous selection criteria. CSOs are the only source of ex-situ conservation for Teak in India. More than 1000 ha of CSOs have been established in India with 450 ha in Maharashtra 240 ha in Madhya Pradesh 120 ha in Karnataka 92 ha in Andhra Pradesh 35 ha in Kerala 30 ha in Orissa and Tamil Nadu and 25 ha in Arunachal Pradesh Katwal 2005 6 . Molecular markers have been looked upon as tools for a large number of applications ranging from localization of a gene to improvement of plant varieties by marker-assisted selection.

slide 2:

260 International Journal of Chemical Studies They have also become extremely popular for phylogenetic analysis adding new dimensions to the evolutionary theories. If we look at the history of the development of these markers it is evident that they have been improved over the last two decades to provide easy fast and automated assistance to scientists and breeders. Genome analysis based on molecular markers has generated a vast amount of information and a number of databases are being generated to preserve and popularize it Joshi et al. 1999 5 . Materials and Methods Clonal Teak Seed Orchard Rajpipla TSO was established in 2000-2001 at Rajpipla in Narmada District for getting good quality seed material. Clonal seed orchard Rajpipla is located at 21 o 53 N Latitude 73 o 31 E Longitude at 45 meters above the mean sea level in Narmada district of South Gujarat in India. The climate of Rajpipla is tropical characterized by fairly hot weather moderately cold winter with humid and warm monsoon coupled with moderately heavy rainfall. The monsoon commenced from second week of June and lasts up to first week of October. Most of the precipitation received from the South-West monsoon during July and August. The average annual rainfall is 1055 mm. Molecular Profile to be subjected to Jaccard’s similarity coefficient analysis using NTSYSpc ver 2.2 software. The genomic DNA extraction protocols was standardized and subjected for analyzing existing molecular variations among Teak clones. Number of monomorphic band Number of polymorphic band and Genetic similarity were recorded. The leaf samples of Tectona grandis L. f. from field plants were collected and stored at - 20 o C. The leaf samples of Tectona grandis L. f. from field plants were collected and stored at -20 o C. The genomic DNA samples extracted from Tectona grandis L. f. leaves were subjected to PCR amplification. Amplification was carried out in a 200 µl thin walled PCR tube containing a 25 µl reaction mix volume. The reaction volume of 25 µl was subjected to amplification through PCR in a thermal cycler Eppendorf along with a control without genomic DNA. Prior to amplification reaction mixture was gently tapped and spun briefly. The genomic DNA amplified using random primers of OPC OPE OPF OPG and OPJ series Operon Tech. California USA. The PCR reactions for RAPD were carried out in a 25 µl of reaction mixture as described by William et al. 1990 12 . The PCR amplification was carried out under following thermal cycling regime The amplified product was collected from the thermal cycler and loaded on to 1.5 percent w/v agarose gel containing 5 µl ethidium bromide prepared in 0.5X TBE pH 8.0. The required volume of 0.5X TBE pH 8.0 was used as running buffer. Whole of the 25 µl PCR amplified product was mixed with 6X gel loading dye of which 15 µl was loaded in well. Along with the samples ‘O’ Range Ruler 500bp ladder ready to use molecular weight premix DNA ladder was also loaded. A potential difference of 5-6 V/cm was provided till the bands resolved properly. The band profiles were visualized and documented using gel documentation system Syngene. For each locus the presence and absence of the polymorphic band was recorded as 0 and 1 respectively. Band positions for each Teak clones and primer combination were scored as either present 1 or absent 0. The scores were entered into a database programme Microsoft Excel and compiled in a binary matrix for phylogenetic analysis using NTSYS-pc Numerical Taxonomy and Multivariate analysis system version 2.2 by Exeter Software Rohlf 2004 7 . The SIMQUALK programme was used to calculate Jaccards similarity coefficient and a graphical phenogram dendrogram of the genetic relatedness among the 15 clones was produced by means of the un-weighted pair group method with arithmetic average UPGMA analysis Sneath and Sokal 1973 9 . Results and Discussion The genomic DNA extracted from each clone was subjected to polymerase chain reaction using random decamers. Initially a total of 200 primers belonging to OPC OPE OPF OPG and OPJ series of universal primers set each consisting of 20 decamers were screened. However primers from OPA and OPL and OPM showed no amplification may be due to absence of complementary sequence in the genome. Finally 7 primers viz. OPC-17 OPE-10 OPE-18 OPF-20 OPG-3 OPG-4 and OPJ-1 were selected for evaluating molecular differences. The nucleotide sequences of each primer are given in Table - 1. It can be seen from Table – 1 that seven decamer primers produced a total of 50 scorable bands in the 15 clones of Tectona grandis L. f. out of which 20 were polymorphic and 30 were monomorphic. The percentage of polymorphism ranged from a maximum 50.00 by OPE-18 to a minimum of 16.67 by OPE-10. Genetic similarity From Table - 2 it is evident that the lowest genetic similarity 0.7209 was between the clones 4 7. The highest genetic similarity 0.9761 was between the clones 9 10. According to dendrogram Fig. - 1 at a similarity level of 70.00 the clones were divided in two main clusters named A and B. Further Cluster-B was divided in to two sub clusters B 1 and B 2 . The clones C 7 and C 8 reported in Cluster-A which were found to be more diverse as compared to other clones. The clones C 1 C 3 C 5 C 2 C 6 and C 4 were observed in sub- cluster-B 1 . Whereas clones C 9 C 10 C 15 C 11 C 12 C 13 and C 14 were registered in sub-cluster-B 2 . The clones C 4 and C 14 found in B 1 and B 2 sub-cluster respectively were having minimum similarity with other clones. Still a screening of more number of primers is recommended to evaluate the present set of clones. Moreover screening of more clones may also give some diverse clones. Several workers have used RAPD markers to detect genetic variation viz. in willow clones in 29 populations of teak Tectona grandis which were collected from central and peninsular India Ginwal et al. 2010 4 in 55 geographically distinct populations of Pinus roxburghii of the Himalayan region Fofana et al. 2008 3 in Tectona grandis grown in Cote d’Ivoire Behera

slide 3:

261 International Journal of Chemical Studies et al. 2008 1 in 38 diverse Indian bitter gourd Mormordica charantia var. charantia and var. muricata accessions Shiran et al. 2007 8 in 39 cultivars and species of almond Syamkumar and Sasikumar 2007 11 in 15 curcuma species Faseela and Joseph 2007 2 in amaranth landraces belonging to three species viz. Amranthus tricolor A. dubius and A. hypochondriacus Zoghlami et al. 2007 13 in 36 Tunisian Opuntia ficus indica L. Mill. ecotypes and Surkhosh et al. 2006 10 in 24 genotypes of Iranian pomegranate Punica granatum L.. Table 1: Details of amplification obtained with different RAPD primers in different clones of Tectona grandis L. f. Primer Number of monomorphic Band Number of polymorphic Band Total number of bands Percentage of polymorphism OPC-17 6 5 11 45.45 OPE-10 5 1 6 16.67 OPE-18 2 2 4 50.00 OPF-20 4 3 7 42.86 OPG-03 5 3 8 37.50 OPG-04 4 1 5 20.00 OPJ-01 4 5 9 55.56 Total 30 20 50 A OPC-17 B OPE-10

slide 4:

262 International Journal of Chemical Studies C OPE-18 D OPF-20 F OPG-3 G OPG-4

slide 5:

263 International Journal of Chemical Studies H OPJ-1 Plate 1: RAPD amplification pattern of different clones of Tectona grandis L. f. by using various series of primer Table 2: Jaccards similarity coefficient among different clones of Tectona grandis L. f. based on the RAPD data C1 C2 C3 C4 C5 C6 C7 C8 C9 C10 C11 C12 C13 C14 C15 C1 1 C2 0.8409 1 C3 0.8809 0.8636 1 C4 0.8139 0.8409 0.8372 1 C5 0.9024 0.8409 0.9268 0.8571 1 C6 0.8181 0.8863 0.8837 0.7777 0.8181 1 C7 0.8048 0.7500 0.7441 0.7209 0.7619 0.6888 1 C8 0.8048 0.7500 0.7441 0.7619 0.8048 0.6888 0.9444 1 C9 0.7777 0.8043 0.8837 0.8181 0.8181 0.7826 0.7674 0.7674 1 C10 0.7608 0.8260 0.8636 0.8000 0.8000 0.8043 0.7500 0.7500 0.9761 1 C11 0.7391 0.7659 0.8409 0.8181 0.8181 0.7446 0.7674 0.8095 0.9523 0.9302 1 C12 0.7826 0.7708 0.8444 0.7826 0.8222 0.7500 0.8139 0.8139 0.9090 0.8888 0.9090 1 C13 0.8409 0.7872 0.9069 0.8000 0.8837 0.8043 0.7500 0.7500 0.8863 0.8666 0.8444 0.9318 1 C14 0.7777 0.8043 0.8837 0.7777 0.8181 0.8222 0.7272 0.6888 0.8636 0.8863 0.8222 0.8666 0.9302 1 C15 0.7234 0.7872 0.8222 0.7608 0.7608 0.7659 0.7906 0.7906 0.9302 0.9534 0.9302 0.9318 0.8666 0.8863 1

slide 6:

264 International Journal of Chemical Studies Fig 1: Dendrogram depicting the genetic relationship among different clones of Tectona grandis L. f. based on RAPD data Conclusion From this molecular variation study it is concluded that the seven decamer primers produced a total of 50 scorable bands in the fifteen clones of Tectona grandis L. f. Out of them 20 were polymorphic and 30 were monomorphic. The percentage of polymorphism ranged from a maximum 50.00 by OPE- 18 to a minimum of 16.67 by OPE-10. The lowest genetic similarity 0.7209 was between the clones 4 7 whereas the highest genetic similarity 0.9761 was between the clones 9 10. The dendrogram of fifteen clones reflected that the clones were divided in two main clusters named A and B. Further Cluster-B was divided in to two sub clusters B 1 and B 2 . The clones C 7 and C 8 reported in Cluster-A which were found to be more diverse as compared to other clones. The clones C 1 C 3 C 5 C 2 C 6 and C 4 were observed in sub-cluster- B 1 . Whereas clones C 9 C 10 C 15 C 11 C 12 C 13 and C 14 were recorded in sub-cluster-B 2 . The clones C 4 and C 14 found in B 1 and B 2 sub-cluster respectively were having minimum similarity with other clones. References 1. Behera TK Singh AK Staub JE. Comparative analysis of genetic diversity in Indian bitter gourd Momordica charantia L. using RAPD and ISSR markers for developing crop improvement strategies. Scientia Horticulture. 2008 115:209-217. 2. Faseela KV Joseph S. Molecular characterization of amaranth land races and assessment of interspecific relationships among Amaranthus spp. L. using RAPD markers. Indian J Genet. 2007 671:12-17. 3. Fofana IJ Ofori E Poitel M Verhaegen D. Diversity and genetic structure of Teak Tectona grandis in its natural range using DNA microsatellite markers. Tropical Conservation Science. 2008 13:279-292. 4. Ginwal HS Chauhan Priti Singh Shalini Maurya Jadon Vikas S. Genetic variability in Pinus roxburghii by RAPD markers. Bioremediation Biodiversity and Bioavailability Global Science Books Ltd. Special 2010 28-34. 5. Joshi SP Prabhakar KR Gupta VS. Molecular markers in plant genome analysis. Plant Molecular Biology Group Division of Biochemical Sciences National Chemical Laboratory Pune India 1999. 6. Katwal RPS. Teak in India: Status prospects and perspectives. In: Bhat K.M. Nair K.K.N. Bhat K.V. Muralidharan E.M. Sharma J.K. eds. Quality timber products of teak from sustainable Forest Management Proceedings of International Conference Kerala Forest Research Institute India and International Tropical Timber Organization Japan 2005 1-17. 7. Rohlf FJ. NTSYS-pc Numerical Taxonomy and Multivariate Analysis System version 2.2 Exeter Software Applied Biostatics New York 2004. 8. Shiran B Amirbakhtiar N Kiani S Mohammadi Sayed- Tabatabaei BE Moradi H. Molecular characterization and genetic relationship among almond cultivars assessed by RAPD and SSR markers. Scientia Horticulture. 2007 111:280-292. 9. Sneath PHA Sokal RR. Numerical Taxonomy. The principle and practice of Numerical Classification. W.H. Freemon and Company San Francisco 1973. 10. Surkhosh A Zamani Z Fatahi R Ebadi A. RAPD marker reveal polymorphism among some Iranian pomegranate Punica granatum L. genotypes. Scientia Horticulture. 2006 111:24-29. 11. Syamkumar S Sasikumar B. Molecular marker based genetic diversity analysis of Curcuma species from India. Scientia Horticulture. 2007 112:235-241. 12. William JGK Kubelik KJ Livak KJ Rafalski JA Tingey SV. DNA polymorphisms amplified by arbitrary primers are useful genetic markers. Nucleic Acid Res. 1990 18:6531-6535. 13. Zoghlami N Chrita I Bouamana B Gargouri M Zemni H Ghorbel A et al. Molecular based assessment of genetic diversity within Barbary fig Opuntia ficus indica L. Mill. in Tunisia. Scientia Horticulture 2007 113:134-141.

authorStream Live Help