Research Journal of Agriculture and Forestry Sciences __________________________________ ISSN 2320-6063 Vol. 1(8), 23-29, September (2013) Res. J. Agriculture and Forestry Sci. International Science Congress Association 23 Genetic Variability and Association Studies among Morphological traits of Leucaena leucocephala (Lam.) de Wit. Genetic ResourcesChavan Sangram1* and Keerthika A.2 National Research Centre for Agroforestry, Jhansi-284003, UP, INDIA Forest College and Research Institute, Tamil Nadu Agricultural University, Mettpalayam-641301, TN, INDIAAvailable online at: www.isca.in Received 18th August 2013, revised 22nd August 2013, accepted 3rd September 2013Abstract Experiment were undertaken to determine variability and correlation among the fifteen Leucaena leucocephala genetic resources from the three states of India viz., Tamil Nadu, Andhra Pradesh and Maharashtra. The observations were recorded on plant height, basal diameter, number of branches and volume index. The fifteen genotypes differed significantly for all the studied traits. The genotypes viz., FCRILL 8 and FCRILL 15 recorded significantly higher plant height, basal diameter and volume index than the rest of the genotypes. The volume index recorded maximum PCV (37.18) and GCV (28.89) and high heritability value of 0.60. The genetic advance as per cent of mean of volume index was 46.23 which were highest among all traits. Volume index exhibited positive and significant phenotypic and genotypic association with plant height (0.76), basal diameter (0.97). Basal diameter exhibited significant phenotypic (0.61) and genotypic (0.77) inter-correlation with plant height. Plant height had positive and non-significant phenotypic (0.02) and genotypic (0.16) inter-association with number of branches. The path analysis study indicated that plant height and basal diameter had direct effect on volume index. Keywords: Genetic variability, genetic advance, heritability, path analysis, genetic resources and Leucaena leucocephala.IntroductionSubabul (Leucaena leucocephala L. de wit) is a miracle tree with a wide assortment of cultivars for different uses like forage, timber, wind break, nitrogen fixation, pulp, etc. for short rotation forestry. Subabul has been introduced in India mainly as an agroforestry crop to meet the increasing demand for fuel, fodder, pulp and timber for poles and posts. It can be grown in variety of soils and climatic conditions due to its tolerance to high temperature and extended drought and remarkable regenerative capacity1,2. The nearly 100 varieties that comprise the species differ markedly in size and form and have been classified into three broad types, the Salvador, the Peru and the Hawaiian. Though the plant has a strong appeal for a multitude of uses, its primary reputation is as forage and feed. But Salvador types like K8, K28 and K67 referred to as Hawaiian giants are suitable for wood production. Leucaena possesses enormous wealth of variability and great potential for economic yield which attract the breeders in utilizing the species in genetic improvement. A logical way to start any improvement programme is to survey the variations present in the germplasm. Hence, the todays challenging task is to screen the naturally available Leucaena leucocephala genetic resources to select the best planting material for higher biomass. Seeds from proven source or plus trees are the backbone of any successful tree improvement and afforestation programme. In order to adopt strategies of conservation and improvement, it is necessary to estimate the amount and distribution of genetic variation in preselected populations of Leucaena leucocephalagenetic resources and it demands intensive research. Most of the economic characters in trees are quantitative in nature. There is a wide range of variability in these characters which depends upon the genetic makeup of the individuals and the environment in which they are grown. Breeders use this variability for achieving improvement in economic characters through efficient selection techniqes4 and knowledge of genetic parameters are required to formulate breeding strategies, as well as estimated breeding values and gains from selection. Thus, the information on genetic parameters such as heritability; genetic advance and genetic correlation is a pre-requisite for making efficient selection strategies by the geneticists and breeders. Due to long gestation period of trees, the analysis of juvenile growths traits is an important technique to establish the relative importance of different genotype as the determinant for improvement. Also Burley and Wood observed that if good correlations exist between measured traits at different stages of the tree’s development, prediction of growth at an advanced age may be possible. The study aims at selection of better genotypes with higher plant height and stem diameter based on correlation with other juvenile growths traits of Leucaena leucocephala. The objectives of this trial was not only to provide a base-population for selection of plus trees, but also to produce improved seeds for new plantings, through converting progeny trials into seedling seed orchards. Most studies in genetic parameters in different tree species were focused on Cotton; Research Journal of Agriculture and Forestry Sciences _______________________________________________ ISSN 2320-6063Vol. 1(8), 23-29, September (2013) Res. J. Agriculture and Forestry Sci. International Science Congress Association 24 Casuarina equisetifolia10; Bombax ceiba; Azadirachta indica11and Simaruba glauca12. In the present study, growth traits such as plant height, collar diameter, number of branches and volume index are used to estimate: (a) differences among the 15 genetic resources and (2) genetic parameters including variability, heritabilities, genetic advance, correlation and path coefficient in a base-population of 15 genetic resources of Leucaena luecocephala. Additionally, this information will be used to discuss the implications and selection strategies of Leucaena for getting higher productivity genotypes to serve as a base for short rotation forestry. Keeping in view the importance of these parameters, the present study was conducted at the Forest College and Research Institute, Mettupalayam and Tamil Nadu News Print Ltd. Karur which are as follows. Material and Methods Genotype collection site: A thorough and extensive wild germplasm exploration survey was undertaken to identify high yielding candidate plus trees (CPTs). Seeds were collected from three states viz., Tamil Nadu, Andhra Pradesh and Maharashtra during the fruiting season of Leucaena in 2008–2009 and planted at captive site of Tamil Nadu News Print Ltd. Karur. The genotypes used in this study are given in figure 1 and table 1. Table-1 Details of Leucaena leucocphala genetic resources and their location Location Name of Assigned District State Latitude Longitude Altitude BAIF, Pune FCRILL 1 Pune Maharashtra 15°06' N 74° 57' E 500 Hyderabad I FCRILL 2 Hyderabad Andhra Pradesh 13° 40'N 79° 20' E 1758 Hyderabad II FCRILL 3 Hyderabad Andhra Pradesh 17° 20'N 78° 30' E 1890 MTP – I FCRILL 4 Coimbatore Tamil Nadu 11 o 20’N 76 o 51’ E 1784 MTP – II FCRILL 5 Coimbatore Tamil Nadu 11 o 20’N 76 o 51’ E 1550 MTP III FCRILL 6 Coimbatore Tamil Nadu 11 o 20’N 76 o 51’ E 1895 MTP IV FCRILL 7 Coimbatore Tamil Nadu 11 o 19’N 77 o 56’ E 987 Coimbatore FCRILL 8 Coimbatore Tamil Nadu 11 o 37’N 77 o 00’ E 1328 Erode I FCRILL 9 Erode Tamil Nadu 11 o 46’N 77 o 00’ E 1149 Erode II FCRILL 10 Erode Tamil Nadu 11 o 46’N 77 o 00’ E 2707 TNPL, Karur FCRILL 11 Karur Tamil Nadu 10 0 58'N 78° 07' E 912 Sivagangai FCRILL 12 Sivagangai Tamil Nadu 9 0 43' N 77Ί 47' E 255 Pudukottai I FCRILL 13 Pudukottai Tamil Nadu 10 0 2' N 77 0 47' E 380 Pudukottai II FCRILL 14 Pudukottai Tamil Nadu 10° 37'N 77° 20’ E 430 Pudukottai III FCRILL 15 Pudukottai Tamil Nadu 9.50' N 78 0 25’ E 358 Figure-1 Distribution of 15 genotypes of Leucaena leucocephala Research Journal of Agriculture and Forestry Sciences _______________________________________________ ISSN 2320-6063Vol. 1(8), 23-29, September (2013) Res. J. Agriculture and Forestry Sci. International Science Congress Association 25 Experimental site and Morphological observation: Field evaluation trial was carried out at Athipalayam which is the captive planting site of Tamil Nadu Newsprint and Paper Ltd. (TNPL), Karur. During 2010-2011, progenies of 15 genotypes were planted in a Randomized Block Design (RBD) with three replications. The 9 plants of fifteen genotypes were planted at 3 x 3 m spacing and are replicated thrice. Observations were recorded on plant height, collar diameter, number of branches and volume index in all the 9 plants (at 9, 12 and 15 months after planting) of each replication. Experimentation and calculation: The data on the four traits were analyzed in RBD. Genetic variability parameters, heritability and genetic advance of the traits were estimated13,14. Replication-wise mean values of the data of each accession were subjected to statistical analysis. The analysis of variance (ANOVA) 15 separates genetic components of variability from total variability. The broad-sense heritability16 were calculated. Phenotypic Coefficient of variation (PCV) and Genotypic Coefficient of Variation (GCV) were estimated using following formula: (Phenotypic Variance) ½ PCV (%) = x 100 General mean (Genotypic Variance) ½ GCV (%) = x 100 General mean Genotypic and phenotypic correlations co-efficients were calculated according to the method suggested by Goulden17. Path co-efficient analysis estimates the direct and indirect effects of each variable by using Deweyand Lu18 methodology. Results and Discussion Growth performance of growth traits: Observation on morphological growth traits (15 months after planting) viz. plant height, basal diameter, number of branches and volume index pertaining to 15 genotypes are presented in table 2. The significant differences were found among the genotypes at 1 per cent level of significance. Only one genotype viz., FCRILL 8 (565.63 cm) at one percent and three genotypes viz., FCRILL 15 (554.97 cm), FCRILL 12 (547.8cm) and FCRILL 7 (545.83 cm) at five per cent level proved superior compared to grand mean (489.90 cm). The highest value of basal diameter was recorded by only one genotype FCRILL 8 (7.97 cm) at one percent level and FCRILL 15 (7.57 cm) at five percent level. A similar pattern in volume index was observed wherein FCRILL 8 and FCRILL 15 put maximum volume index. Among the 15 genotypes, two genotype viz., FCRI LL 8 and FCRI LL 15 proved to be a good performer which expressed superiority for all the four characters viz., height, basal diameter, volume index and number of branches investigated. A plethora of workers reported the existence of significant differences and superiority of few seed sources, progenies and provenances in various tree species like Acacia nilotica19Cordia alliodora20; Dalbergia sissoo21 which lend support to the current findings in Leucaena genetic resources. Variability, heritability and genetic advance studies in growth attributes: The perusal of data revealed significant genetic variation among 15 genotypes of Leucaena leucocephala for growth attributes (table 3). Volume index registered highest Phenotypic co-efficient of variation (PCV) (37.18%) followed by basal diameter, number of branches and plant height. Volume index also registered high genotypic co-efficient of variation (GCV) (28.89%) followed by rest of the traits. The high estimates of heritability observed for number of branches (0.61) followed by volume index (0.60) and moderate heritability for height (0.55) and basal diameter (0.46). High degree of genetic advance as per cent of mean was observed for volume index (46.23%) followed by number of branches, basal diameter and plant height. The extent of variability was measured by GCV and Phenotypic co-efficient of variation (PCV) which provides information about relative amount of variation in different character22. In the current study; volume index registered the high PCV, GCV, heritability and genetic advance. The GCV was found to be low magnitude than the PCV for all the studied traits. This indicates that these traits are influenced by the local environmental factors as evidenced in Acacia nilotica23 neem11, Dalbergia sissoo24 and also in the progenies of Melia dubia25 which lend supports to the results of current investigation. The heritability states the magnitude of inheritance of quantitative traits while genetic advance provide needful information for formulating suitable selection procedure22. The knowledge of heredity and environment to the fullest extent will give opportunity to understand the role of heredity among quantitative traits26. In the present study, the high heritability with high genetic gain was observed for volume index. Hence, the higher heritability recorded in volume index, so it could be a reliable indicator for further improvement programme. High heritability coupled with high genetic advance indicated that such characters are controlled by additive gene action and it will responds effectively on phenotypic selection. The findings of current study are in line with those of Ramachandra27 in Acacia catechu and Devagiri28 in Heracleum Candicans. Association Studies in growth attributes of Leucaena: Correlation studies: Volume index exhibited positive and significant phenotypic association with plant height (0.76), basal diameter (0.97). It had significant genotypic correlation with plant height (0.89) and basal diameter (0.98). Basal diameter exhibited significant phenotypic (0.61) and genotypic (0.77) inter-correlation with plant height. Plant height exhibited positive and non-significant phenotypic (0.02) and genotypic (0.16) inter-association with number of branches (table 4). In the present investigation, degree of correlation co-efficient at genotypic level was higher than their corresponding phenotypic co-efficient of correlations in all the parameters indicating the genetic association among the character. The correlation Research Journal of Agriculture and Forestry Sciences _______________________________________________ ISSN 2320-6063Vol. 1(8), 23-29, September (2013) Res. J. Agriculture and Forestry Sci. International Science Congress Association 26 coefficient values at genotypic level were higher than phenotypic coefficient of correlations in all the parameter, which indicates the genetic association among the characters. This result is an agreement with the findings of Pongamia pinnata.26 In the current study, significant and positive correlations were observed for volume index with plant height and basal diameter. The results are in agreement with earlier findings in Casuarina equisetifolia10. Plant height had positive and significant correlation with basal diameter. Similar result was reported in Bauhinia variegata29. The present investigations indicated that basal diameter and plant height could reliably serve as selection criteria to increase wood volume in Leucaena for pulp and energy wood production. Path analysis: Though correlation analysis measures the relationship between dependent and independent characters, it does not provide exact picture of how much a character constitute on its own and via other characters as the dependent trait. Under such circumstances, path coefficient analysis is helpful in the resolution of correlation into components of direct and indirect effects. Among the traits studied, plant height (0.33) and basal diameter (0.73) exercised higher positive direct effect on volume index. But, the number of branches (-0.017) exerted negative direct effect on volume index. The plant height exerted positive indirect effect via basal diameter (0.56) and number of branches (0.05). Basal diameter recorded positive and moderate indirect effect via plant height (0.25) and number of branches (0.19) on volume index (table 5). Path analysis gives an insight into a complex relationship between different characteristics in a biological system30. In the present investigation, the plant height and basal diameter exercised positive direct effect on volume index and plant height exerted positive indirect effect via basal diameter and number of branches. Similarly, basal diameter recorded indirect effect viaplant height on volume index which indicated major contribution towards volume. Hence from the current study, high and positive association with intensive direct effect of plant height and basal diameter on volume index could be used as valuable and reliable measure for Leucaena leucocephala tree improvement programme. Wide range of research findings in different tree species viz., Simarouba glauca12 and Terminalia arjuna31 also reported similar results these extend support to the results of current study. Table-2 Variation in growth attributes for Leucaena leucocephala genetic resources (15 MAP) Genotypes Plant Height Basal Diameter Number of branches Volume index FCRI LL 1 421.37 5.79 6.24 14118.26 FCRI LL 2 480.63 5.52 5. 25 14617.08 FCRI LL 3 450.26 5.74 6.34 14891.2 FCRI LL 4 463.89 6.68 5.52 21419.34 FCRI LL 5 481.2 6.14 6.12 18344 FCRI LL 6 499.42 5.36 5.82 14898.98 FCRI LL 7 545.83* 6.41 5.61 22633.33 FCRI LL 8 565.63** 7.97** 7.16* 35912.93** FCRI LL 9 443.7 5. 99 6.31 16272.46 FCRI LL 10 460.87 6.24 5.20 18569.49 FCRI LL 11 433.78 6.17 6.03 15473.08 FCRI LL 12 547.8* 7.59* 5.10 31785.72** FCRI LL 13 480.94 6.25 5.40 20267.38 FCRI LL 14 518.15 6.81 7.41** 24141.33 FCRI LL 15 554.97* 7.57* 7.02* 31850.07** Mean 489.90 6.42 6.01 21012.98 SEd 26.59 0.49 0.49 3552.35 CD (0.01) 73.50 1.35 1.35 9817.53 (* at 5% significant level, ** at 1% significant level)Table-3 Genetic estimates for growth attributesTraits GCV (%) PCV (%) Heritability (%) Genetic advance (%) Height 7.77 10.49 0.55 11.85 Basal diameter 10.10 14.93 0.46 14.10 No. of branches 9.23 11.88 0.61 14.95 Volume index 28.89 37.18 0.60 46.23 Research Journal of Agriculture and Forestry Sciences _______________________________________________ ISSN 2320-6063Vol. 1(8), 23-29, September (2013) Res. J. Agriculture and Forestry Sci. International Science Congress Association 27 Table-4 Phenotypic and genotypic correlation coefficient of growth attributes Traits Plant height Basal diameter No. of branches Volume index Plant height P 1.000 0.61** 0.020 0.76** G 1.000 0.77** 0.16 0.89** Basal diameter P 1.000 0.172 0.97** G 1.000 0.26 0.98** No. of branches P 1.000 0.17 G 1.000 0.23 **Significant at 1% level. Table-5 Path coefficient analysis of morphometric traits on volume index Traits Plant height Basal diameter No. of branches Correlation (r) with Volume index Plant height 0.331 0.563 -0.003 0.331 Basal diameter 0.254 0.732 -0.05 0.732 No. of branches 0.054 0.193 -0.017 -0.017 Figure 2 Path diagram showing relationship between volume index and morphometric traits (15 MAP)Conclusion From the current study, FCRILL 8 and FCRILL 15 recorded superiority in terms of growth characteristics viz., plant height, basal diameter, number of branches and volume index. These two genotypes can be exploited for future improvement programmes. Also the present investigation envisaged that high and positive association coupled with intensive direct effect by plant height and basal diameter on volume index could be used as valuable, reliable and relevant yardstick for Leucaena breeding programme. Hopefully genetic knowledge of tree species will be helpful to propagators, geneticists, breeders and tree improvement specialists in maximizing the quality and productivity of the plantation to meet their market demand over a time. Research Journal of Agriculture and Forestry Sciences _______________________________________________ ISSN 2320-6063Vol. 1(8), 23-29, September (2013) Res. J. Agriculture and Forestry Sci. International Science Congress Association 28 Acknowledgements The authors are thankful to ICAR-NAIP Scheme and Forest College and Research Institute, Mettupalayam who supported for research activities. Mention is also merited to Dr. K.T. Parthiban, Professor and head, Department of Tree Breeding for his supervision, advice, and guidance during study. References 1.Devaranavadgi S.B., Murthy B.G., Desai S.A. and Yelashetty S., Preliminary evaluation of subabul genotypes for psylid tolerance, Cur. Res.,25, 64-65 (1996)2.Devaranavadgi S.B., Guggari A.K., Kalaghatagi S.B., Wali S.Y. and Nadagoudar S.B., Suitable Leucaena genotypes for shallow black soils of northern dry zone of Karnataka, Annals of Arid Zone,38(1), 45-48 (1999)3.Pathak P.S and Patil B.D., Leucaena Research at the IGFRI. In: Leucaena research in the Asian – Pacific Region. Proc. Workshop, Singapore, Nov. 23-26, NFTA & IDRC (1982)4.Safavi S.M., Farshadfar M.D., Kahrizi and Safavi S.A., Genetic variability of some morphological traits in Poplar Clones, American Journal of Scientific Research,13, 113-117 (2011)5.Wright S. Correlation and causation, J. Agric. Res., 20,557-585 (1921)6.Lewis A., Pande K.K., Salil K. Tewari, Gahalain S.S. and Pankaj Sah, Genetic Variability for different traits in the Indian Bamboo, New York, Science Journal,3(6), 27-31 (2010)7.Chaturvedi O.P. and Pandey N., Correlation and Path Analysis Studies between Biomass and Other Characters in Bombax ceiba L., Silvae Genetica, 53(5–6), 269-271 (2005)8.Burley J. and Wood P.J., A manual on species and provenance research with particular reference to the tropics, Tropical Forestry Papers,10, 34-61 (1978)9.Singh R.B. and Gupta M.P., Multivariate analysis of divergence in upland cotton, Indian Journal of Genetics,28, 151-157 (1968) 10.Ashok Kumar and Paramathma M., Correlation and path coefficient studies in Casuarina equisetifolia L. Johnson, Indian Forester, 131(1), 47-55 (2005) 11.Dhillon R.S., Bisla S.S., Arya S. and Hooda M.S., Genetic variation, heritability and correlations for growth parameters in Azadirachta indica. A. Juss, Ann. For.,11(2), 215-221 (2003)12.Kumaran K., Nesamani K. and Govinda Rao M., Correlation and Path coefficient studies in Simaruba glauca DC. Indian Forester, 2, 322-330 (2010)13.Johnson H.W., Robinson H.F. and Comstock R.E., Genotypic and Phenotypic correlations on soyabean and their implications in selection, Agron. J., 47, 477-483 (1955)14.Nya E.J. and Eka M.J., Genetic variability and heritability studies of desirable metric characters in Talinum triangulare land races in South Eastern Nigeria, J. Agron.,6, 459-463 (2007)15.Panse V.G. and Sukhatme P.V., Statistical methods for Agricultural workers, ICAR Publication, New Delhi, 337 (1978)16.Lush K.I., Intrasite correlation and regression of spring on dams as a method of establishing heritability of characters, Proc. Amer. Soc. Animal Production, 33, 293-301 (1940)17.Goulden C.H., Some distance properties of latent root and vector methods used in multivariate analysis, Biometrika,53, 325-338 (1952)18.Dewey R.D. and Lu K.H., A correlation and path analysis components of crested wheat grass seed production, Agron. Journal, 51, 515-518 (1959)19.Padmini S. and Banerjee A.C., Provenance trails of Acacia nilotica,J. Tree Sci., 5, 53-56 (1986) 20.Lahiri A.K., Trials of Cordia alliodora provenances in North Bengal, Indian Forester 114, 51-53 (1988)21.Tewari S.K., Kohli R.K., Arya K.S. and Atal, Diagnostic survey of Shisham. In proceedings, IUFRO-DNAES International Meeting: Resource Inventory Techniques to Support Agroforestry and Environment, 53-55 (1996)22.Sumathi P., Sumanth M., and Veerabadhiran P., Genetic variability for different biometrical traits in pearl millet genotypes (Pennisetum glaucum L. R. BR.), ElectronicJournal of Plant Breeding, 1(4), 437-440 (2010)23.Ginwal H.S. and Mandal A.K., Variation in growth performance of Acacia nilotica Willd. ex Del. Provenances of wide geographical origin : Six Year results, Silvae Genetica, 53, 5–6 (2004)24.Dogra A.S., Nautiyal S., Nautiyal D.P. and Singh G., Evaluation of field performance of 34 progenies of Dalbergia sissoo in Punjab, Ann. For., 13(2), 199-204 (2005)25.Kumar P., Parthiban K.T., and Sarvanan V., Genetic Variations among Open Pollinated Families of Selected Better Trees in Melia Dubia, Research Journal of Recent Sciences,2(ISCA-2012), 189-194 (2013)26.Rao G.R., Shankar A.K., Srinivas I., Korwar S.R. and Venketesh V., Diversity and variability in seed characters and growth of Pongamia pinnata (L.) Pierre accessions, Trees, 25, 725–734 (2011) Research Journal of Agriculture and Forestry Sciences _______________________________________________ ISSN 2320-6063Vol. 1(8), 23-29, September (2013) Res. J. Agriculture and Forestry Sci. International Science Congress Association 29 27.Ramachandra N.G., Provenance variation in seed and seedling parameters in Acacia catechu.Willd. Ph.D. Thesis, FRI Deemed University, Dehra Dun (1996)28.Devagiri G.M., Evaluation of seed source variation in seed and seedling traits in Dalbergia sissoo Roxb. Ph.d. Thesis,Forest Research Institute University, Dehra Dun, India (1997)29.Thakar I.K., Seed source evaluation for growth and nutrient parameters of Bauhinia variegate,Indian J. of agroforestry,12(2), 96-100 (2010)30.Pandey Pandey V., Tripathi S. and Tewari S.K., Path analysis in Populus deltoides Bart. 8 year age, Indian Forester,123(8), 755-758 (1997)31.Srivastava D.P., Srivastav P.K., A.K. Goel and Thangavelu K., Correlation and path coefficient studies in Terminalia arjuna,Ann. For., (1), 178-181(1993)