Scientia Horticulturae 148 (2012) 230234

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Scientia Horticulturae
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Characterization and evaluation of genetic diversity of Iranian mango (Mangifera indica L., Anacardiaceae) genotypes using microsatellites
M. Shamili a , R. Fatahi b , J.I. Hormaza c,
a b c

Departure of Horticulture Science, Hormozgan University, Iran Departure of Horticulture Science, Tehran University, Iran Instituto de Hortofruticultura Subtropical y Mediterrnea La Mayora (IHSM-UMA-CSIC), 29750 Algarrobo-Costa, Mlaga, Spain

a r t i c l e

i n f o

a b s t r a c t
Mango is an important tropical fruit crop that is believed to be native to Southeastern Asia and currently cultivated worldwide in regions with tropical and subtropical climates. However, the extant diversity in most mango producing countries is still poorly understood. In this work we have used 16 Simple Sequence Repeat (SSR) markers to analyze 41 mango genotypes present in Iran. A total of 56 alleles were detected from 15 polymorphic loci ranging from 2 to 6 alleles per locus with an average of 3.7 alleles per locus. Mean expected and observed heterozygosities over the 15 polymorphic SSR loci averaged 0.57 and 0.63 respectively. The SSRs studied allowed the unambiguous identication of all the mango genotypes analyzed except few synonymies. UPGMA and Bayesian cluster analysis suggested a clear separation between the Iranian cultivars originated from India and Pakistan. 2012 Elsevier B.V. All rights reserved.

Article history: Received 10 August 2012 Received in revised form 25 September 2012 Accepted 26 September 2012 Keywords: Fingerprinting Genetic diversity Genetic structure Molecular markers SSRs Tropical fruits

1. Introduction Mango (Mangifera indica L., 2n = 40) is one of the approximately 850 species classied into 73 genera in the Anacardiaceae (order Sapindales), a family of mainly tropical species with a few representatives in temperate regions (Bompard, 2009). This family includes other cultivated species such as pistachio (Pistacia vera L.), cashew (Annacardium occidentale L.), ambarella (Spondias dulcis Forst.) or yellow and purple mombins (Spondias mombin L. and Spondias purpurea L., respectively). The genus Mangifera contains about 70 species mostly restricted to tropical Asia and can be divided into two subgenera (Limus and Mangifera) (Bompard, 2009; Kostermans and Bompard, 1993) with mango belonging to the subgenus Mangifera. Although some authors have considered India as the center of origin due to the high degree of mango diversity observed in that country (Ravishankar et al., 2000), taxonomic and molecular evidence also supports an evolution of mango within a larger area including northwestern Myanmar, Bangladesh and Northeastern India (Bompard, 2009; Mukherjee, 1972; Mukherjee and Litz, 2009). In any case, it is likely that mango cultivation originated

Corresponding author. Tel.: +34 952548990; fax: +34 952552677. E-mail address: ihormaza@eelm.csic.es (J.I. Hormaza). 0304-4238/$ see front matter 2012 Elsevier B.V. All rights reserved. http://dx.doi.org/10.1016/j.scienta.2012.09.031

in India where over 1000 varieties are recognized, most of them selections from naturally occurring open-pollinated seedlings (Iyer and Degani, 1997). Mango cultivation spread outside its center of origin and domestication throughout many tropical and subtropical regions of the world along trading routes where selections of genotypes best adapted to particular conditions were made (Bompard, 2009; Lpez-Valenzuela et al., 1997; Mukherjee and Litz, 2009) reaching Iran at least about 400 years ago (Samavi and Saiedi, 1991) although Persian traders could have introduced the crop much earlier, around the 10th century (Mukherjee and Litz, 2009). Currently, mango is produced in more than 90 countries worldwide and, although only a small proportion of the production enters international trading channels, the volume traded has increased signicantly in the last two decades (Evans and Mendoza, 2009). Total world mango production has reached nearly 40 million tons in 2010 making mango one of the ve most important fruit crops worldwide (after bananas, oranges, grapes and apples). A few countries (India, China, Thailand, Mexico, Pakistan and Indonesia) account for over 75% of world production, being India the main producer with over 40% (over 15 millions of tons) (FAOSTAT, 2012). Traditional characterization of mango cultivars has been made based on phenological and morphological traits in owers, leaves, fruits and seeds (IPGRI, 2006; Knight et al., 2009). However, although morphological characterization is still necessary for proper genotype characterization, cultivar identication based exclusively on phenotypic traits is inaccurate due to the inuence

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of the environment and phenology and the often limiting number of discriminating traits. Thus, molecular markers provide an attractive and more reliable alternative to morphological markers. As a consequence, recently, as in other fruit tree species (Wnsch and Hormaza, 2002), molecular identication of mango cultivars has been carried out with different molecular systems as isozymes (Degani et al., 1990), minisatellites (Adato et al., 1995), ISSRs (Eiadthong et al., 1999; Samal et al., 2012), AFLPs (Eiadthong et al., 2000; Kashkush et al., 2001), RAPDs (Hemanth Kumar et al., 2001; Karihaloo et al., 2003; Lpez-Valenzuela et al., 1997; Ravishankar et al., 2000; Samal et al., 2012; Schnell et al., 1995), SCoTs (Luo et al., 2010) and, more recently, microsatellites or SSRs (Chiang et al., 2012; Duval et al., 2005; Hirano et al., 2010; Honsho et al., 2005; Ravishankar et al., 2011; Schnell et al., 2005; Viruel et al., 2005). Similarly to other countries where mango has been introduced, the diversity of mangoes present in Iran has been poorly studied. In order to ll this gap, in this work, a total of 16 SSRs were used to ngerprint and examine the genetic diversity of 41 mango genotypes present in Iran. 2. Materials and methods 2.1. Plant material and DNA extraction In this study, 41 mango genotypes, sampled from three different areas of Hormozgan province (Minab, Roodan and Siyahoo) in Southern Iran, where most of the mango production in the country takes place, were analyzed (Table 1). These three regions differ from each other in the average temperature, relative humidity and altitude with Minab showing the warmer and more humid climate, Roodan is the cooler area and Siyahoo climate is in between the other two. Total genomic DNA was extracted from young leaves as described by Murray and Thompson (1980). 2.2. SSR analysis

are the frequency of the ith and jth alleles respectively) that measures the probability that two randomly drawn diploid genotypes will be identical assuming observed allele frequencies and random assortment (Paetkau et al., 1995). The program ARLEQUIN version 3.01 (Excofer et al., 2005) was used to calculate A, Ho and He . POPGENE 1.32 software (Yeh et al., 1997) was used to calculate Ne and F. PI was calculated by IDENTITY 1.0 (Centre for Applied Genetics, University of Agricultural Sciences, Vienna, Austria). The genetic relationships among the accessions studied were calculated using UPGMA cluster analysis of the similarity matrix obtained from the proportion of shared amplication fragments (Nei and Li, 1979) with NTSYSpc 2.11 (Exeter Software, Stauket, NY, USA). The cophenetic correlation coefcient was computed for the dendrogram after the construction of a cophenetic matrix to measure the goodness of t between the original similarity matrix and the dendrogram. In order to analyze with more detail the relationships between the different genotypes studied, assignation of the genotypes to different putative populations was studied with the program STRUCTURE version 2.3.4 (Falush et al., 2003; Hubisz et al., 2009; Pritchard et al., 2000) that identies clusters of individuals on the basis of their genotypes at multiple loci using a Bayesian approach maximizing HardyWeinberg equilibrium within each cluster. We used the admixture option (where each individual can have some fraction of its genome from ancestors in other clusters) and performed several runs of various lengths to infer the number of genetic clusters (k) represented by the individuals genotyped, testing all values of k from 1 to 10. The k value that provided the maximum likelihood over the runs was retained as the most probable number of clusters (Pritchard et al., 2010). Clustering solutions of the highest likelihood were obtained when most genomic assignments were distributed over 24 clusters. For those, each k was simulated 10 times and was run for 200,000 steps, after a burn-in period of 20,000 steps (Falush et al., 2007; Hubisz et al., 2009).

3. Results and discussion A total of 16 SSR loci published previously (Viruel et al., 2005) were used in this analysis. DNA amplication was carried out following the protocol of Viruel et al. (2005). Amplication reactions were performed in 15 L volumes containing 16 mM (NH4 )2 SO4 , 67 mM TrisHCl pH 8.8, 0.01% Tween20, 2 mM MgCl2 , 0.1 mM each dNTP, 0.4 M each primer, 25 ng genomic DNA and 1 unit of BioTaqTM DNA polymerase (Bioline, London, UK) on an I-cycler (Bio-Rad Laboratories, Hercules, CA, USA) thermocycler using the following temperature prole: an initial step of 1 min at 94 C, 35 cycles of 30 s at 94 C, 30 s at 55 C and 1 min at 72 C, and a nal step of 5 min at 72 C. Forward primers were labeled with a uorescent dye on the 5 end (Proligo, Paris, France). The PCR products were analyzed by capillary electrophoresis in a CEQTM 8000 capillary DNA analysis system (Beckman Coulter, Fullerton, CA, USA). Samples were denaturalized at 90 C during 120 s, injected at 2.0 kV 30 s and separated at 6.0 kV during 35 min. Each reaction was repeated at least twice in each run to ensure size accuracy and to minimize run-to-run variation. 2.3. Data analysis Allelic composition of each accession and the number of total alleles was determined for each SSR locus. The genetic information was assessed using the following parameters: number of alleles per locus (A), effective number of alleles (Ne = 1/1 He ), observed heterozygosity (Ho , direct count), expected heterozygosity (He = p2 where pi is the frequency of the ith allele, Nei, 1973), 1 i Wrights xation index (F = 1 Ho /He ) (Wright, 1951) and the probp4 + (2pi pj )2 , where pi and pj ability of identity (PI = 1 i 3.1. SSR polymorphism and genetic diversity From the 16 loci initially analyzed, only one (LMMA5) was monomorphic across all the genotypes studied; the rest produced polymorphic amplication fragments and amplied single loci in the cultivars studied. The diversity parameters for the 15 polymorphic single-locus SSRs are shown in Table 2. The analysis of those 15 SSRs in 41 mango genotypes from Iran detected a total of 56 polymorphic bands, with an average of 3.7 bands/SSR, ranging from of 2 to 6 bands/SSR. The effective number of alleles ranged between 1.07 and 2.71 with an average of 1.51. Fifteen unique alleles were observed in the genotypes studied, mostly in genotypes from Pakistan. Thus, in Langra1 two specic alleles were observed in LMMA6 and one at each of LMMA3, LMMA9, LMMA11, LMMA13 and LMMA16; in Senderi2, two specic alleles were observed in each of LMMA2 and LMMA10 loci; in Senderi1 two specic alleles were observed in LMMA10; nally, in Khooshei (a genotype of unknown origin) one specic allele was observed at each of LMMA2 and LMMA3 loci. Observed heterozygosity ranged from 0.30 (LMMA14) to 0.87 (LMMA3) (mean 0.63), whereas expected heterozygosity ranged from 0.29 (LMMA14) to 0.84 (LMMA3) (mean of 0.57). Expected and observed heterozygosity values were compared using the xation index (F), which had an average over all the SSR loci of 0.10 with values between 0.01 (LMMA10 and LMMA16) and 0.41 (LMMA6). For all the loci this parameter was negative indicating an excess of heterozygotes in the genotypes analyzed. Regarding the probability of identity, the maximum (0.55) was detected in

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Table 1 List and characteristics of the 41 mango genotypes analyzed in this work. Genotype Langra1 Senderi1 Helov Anonym1 Doodoo Almehtar1 Arbabi Zarak Majlesi Anonym2 Moshk Anonym3 Kalak1 Charak1 Khiyar1 Haji Senderi2 Anonym4 LangraB9 Khooshei Mikhaki1 Kiloo Mikhaki2 Nabati2 Belersan Almehtar2 Shahani Anonym5 Anonym6 Anonym7 Anonym8 Anonym9 Kalak2 Charak2 Khiyar2 Kalak3 Gol Khodroo Anonym12 Nessa Zapak Sampling region in Iran Minab Minab Minab Minab Minab Minab Minab Minab Minab Minab Minab Minab Minab Minab Minab Minab Minab Minab Roodan Roodan Roodan Roodan Roodan Roodan Roodan Roodan Roodan Roodan Roodan Roodan Roodan Roodan Roodan Roodan Roodan Roodan Siyahoo Siyahoo Siyahoo Siyahoo Siyahoo Reported geographic origin Pakistan Pakistan Unknown India India Unknown India India India India India India India India India India Pakistan India Pakistan Unknown India India India India India India India India India India India India India India India India India India India India India Fruit shape Elongated Elongated Oblong Oblong Oblong Oblong Oblong Oblong Oblong Oblong Oblong Oblong Oblong Oblong Oblong Oblong Elongated Oblong Elongated Oblong Oblong Oblong Oblong Oblong Oblong Oblong Oblong Oblong Oblong Oblong Oblong Oblong Oblong Oblong Oblong Oblong Oblong Oblong Oblong Oblong Oblong Fruit size Large Large Medium Medium Medium Medium Medium Medium Medium Medium Medium Medium Medium Medium Medium Medium Large Medium Large Medium Medium Medium Medium Medium Medium Medium Medium Medium Medium Medium Medium Medium Medium Medium Medium Medium Medium Medium Medium Medium Medium Pulp color Orange/yellow Orange Yellow Yellow Yellow Yellow Yellow Yellow Yellow Yellow Yellow Yellow Yellow Yellow Yellow Yellow Orange Yellow Orange/yellow Yellow Yellow Yellow Yellow Yellow Yellow Yellow Yellow Yellow Yellow Yellow Yellow Yellow Yellow Yellow Yellow Yellow Yellow Yellow Yellow Yellow Yellow Pulp ber Absent Absent Present Present Present Present Present Present Present Present Present Present Present Present Present Present Absent Present Absent Present Present Present Present Present Present Present Present Present Present Present Present Present Present Present Present Present Present Present Present Present Present Total soluble solids to acid ratio High High Low Medium Low Low Medium Medium Medium Medium Medium Medium Low Low Low Low High Low High Medium Medium Medium Medium Medium Medium Medium Medium Medium Medium Medium Medium Medium Medium Medium Medium Medium Medium Medium Medium Medium Medium

Table 2 Diversity parameters using of 15 SSR loci in 41 Iranian mango genotypes. SSR LMMA1 LMMA2 LMMA3 LMMA4 LMMA6 LMMA7 LMMA8 LMMA9 LMMA10 LMMA11 LMMA12 LMMA13 LMMA14 LMMA15 LMMA16 Average Number of alleles (A) 3 4 6 3 6 3 3 3 3 5 3 3 2 5 4 3.7 Number of effective alleles (Ne ) 2.09 1.07 1.51 1.89 2.09 1.28 1.21 1.07 1.05 2.71 1.89 1.13 1.27 1.19 1.19 1.51 Observed heterozygosity (Ho ) 0.68 0.48 0.87 0.53 0.55 0.59 0.53 0.73 0.62 0.83 0.51 0.64 0.30 0.79 0.76 0.63 Expected heterozygosity (He ) 0.65 0.39 0.84 0.47 0.39 0.54 0.50 0.71 0.61 0.61 0.47 0.51 0.29 0.77 0.75 0.57 Fixation index (F) 0.04 0.23 0.03 0.12 0.41 0.09 0.06 0.03 0.01 0.30 0.08 0.25 0.03 0.02 0.01 0.10 Probability of identity (PI) 0.19 0.21 0.15 0.22 0.13 0.26 0.22 0.55 0.13 0.15 0.15 0.50 0.13 0.19 0.19 0.22

LMMA9 with 3 alleles and the minimum (0.13) in LMMA6, LMMA10 and LMMA14 with 6, 3 and 2 alleles, respectively. The average was of 0.22. The fact that the average F was close to 0 suggests that the group of genotypes studied shows a global behavior similar to a random mating population. Probably this can be explained because most, if not all, the genotypes studied are the result of selection from open-pollinated chance seedlings resulting from

natural cross-pollinations. In Iran most of the mango genotypes are of unknown pedigree and have been propagated by seed (Shamili et al., 2011). The results obtained in this work can be compared with those of Viruel et al. (2005) who used the same set of loci and Hirano et al. (2010) who used some of the loci used here but both works analyzed cultivars from different geographical origins and genetic

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Fig. 1. Dendrogram of the 41 mango genotypes studied based on UPGMA analysis using the similarity matrix generated by the Nei and Li coefcient with 56 SSR fragments (a) and bar plot made by Structure at k = 2(b), k = 3(c) and k = 4(d). Each individual is represented by a horizontal colored line.

backgrounds. Thus, the number of alleles obtained in our work (average of 3.7) is lower than that of Viruel et al. (2005) (average of 5.3) and Hirano et al. (2010) (average of 7.55) probably due to the fact that the genotypes analyzed in our work are just from Iranian origin. In spite of the probably close genetic relationships among most of the cultivars studied, relatively high values of observed (0.62) and expected (0.58) heterozygosities were found compared to those reported by Viruel et al. (2005) (0.69 and 0.65, respectively) and Hirano et al. (2010) (0.64 and 0.71, respectively); this, together with the low PI value observed (0.22), indicates that a relatively high variability is present in the genotypes studied and, consequently, that the Iranian germplasm can be valuable to increase the genetic base of cultivated mango. 3.2. Genetic relationships among Iranian mango genotypes The dendrogram obtained after UPGMA analysis of the 56 amplication fragments detected with the 15 SSRs used, shown in Fig. 1a, allowed an unambiguous discrimination of most of the 41 Iranian mango genotypes studied with some synonymies, such as Anonym1 and Mailesi, Khodroo and Anonym12, Moshk, Anonym6 and Charak2 or Kalak2 and Kalak3. Most accessions with similar names (except Kalak2 and Kalak3) showed different banding patterns for the SSR loci studied, indicating that they could be homonymies. A single tree was obtained with a high cophenetic correlation coefcient between the cophenetic matrix and the similarity matrix (0.94) reecting a good t between the cophenetic and the similarity matrixes. The same genotypes were also grouped by Bayesian clustering analysis using the STRUCTURE software, that uses the allele information per locus instead of the presence or absence of fragments described above. Clustering solutions of the highest likelihood were obtained when most genomic assignments were distributed over 24 clusters, although the highest likelihood was observed for k = 2. We examined with more detail the results obtained with k = 2 (Fig. 1b), k = 3 (Fig. 1c) and k = 4 (Fig. 1d). With k = 2, there is a large cluster of genotypes of reported Indian origin and a second cluster with the genotypes from Pakistan; in fact, LangraB9 that is grouped with the Indian genotypes after UPGMA analysis, clearly belongs to the same population as the rest of the Pakistanian genotypes according to the Bayesian clustering analysis. When

k is increased to 3, the Pakistanian cluster almost does not change and the group of Indian origin seems to be formed by a mixture of two different genetic backgrounds. With k = 4, both the Pakistanian and Indian clusters seem to be formed by a mixture of two different genetic backgrounds but the clear differentiation between the Pakistanian and Indian clusters is maintained. Consequently, k = 2 (e.g. two populations, Pakistanian and Indian) is not only the most statistically consistent but also the most parsimonious k based on the outputs generated by the STRUCTURE software. The four accessions introduced into Iran from Pakistan (Langra1, Langra B9, Senderi1 and Senderi2) and three accessions of unknown origin (Helov, Almehtar1 and Khooshei) are clearly separated from the rest. They are also different in fruit shape, color, taste and size compared with the rest of the genotypes (Shamili et al., 2011) and, consequently, could probably be derived from the same genetic background. The remaining 34 genotypes, from Indian and unknown origins, have smaller fruits, lower sugar to acid ratio and more bers in the pulp (Shamili et al., 2011). LangraB9 was assumed to be the same genotype as Langra1 by producers, although the results obtained in this work suggest that they are indeed different genotypes. The results presented in this work provide a detailed picture of genetic diversity of mango germplasm in Iran. Our results conrm a high diversity of mangoes cultivated in Iran revealing a mixed pattern of genotypes, although clearly separating the accessions originated from India from those from Pakistan, that could be the result of the long history of mango cultivation in this country. Molecular markers, such as microsatellites, are powerful tools that should be used to optimize germplasm management and standardize the nomenclature of mango cultivars in the different mango growing regions due to the high number of synonymies/homonymies present in order to have a complete picture of mango germplasm diversity.

Acknowledgements The research was supported by the Spanish Ministry of Science European Regional Development Fund, European Union (AGL201015140) and the Agriculture and Natural Resources Faculty of Tehran University.

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M. Shamili et al. / Scientia Horticulturae 148 (2012) 230234 Knight Jr., R.J., Campbell, R.J., Maguire, I., 2009. Important mango cultivars and their descriptors. In: Litz, R.E. (Ed.), The Mango, 2nd Edition: Botany, Production and Uses. CAB International, Wallingford, UK, pp. 4266. Kostermans, A.J.G.H., Bompard, J.M., 1993. The Mangoes, Their Botany, Nomenclature, Horticulture and Utilization. IPGRI. Academic Press. Lpez-Valenzuela, J.A., Martnez, O., Paredes-Lpez, O., 1997. Geographic differentiation and embryo type identication in Mangifera indica L. cultivars using RAPD Markers. HortScience 32, 11051108. Luo, C., He, X., Chen, H., Hu, Y., Ou, S., Gao, M.P., 2010. Analysis of diversity and relationships among mango cultivars using Start Codon Targeted (SCoT) markers. Biochem. Syst. Ecol. 38, 11761184. Mukherjee, S.K., 1972. Origin of mango (Mangifera indica L.). Econ. Bot. 26, 260266. Mukherjee, S.K., Litz, R.E., 2009. Introduction: botany and importance. In: Litz, R.E. (Ed.), The Mango, 2nd Edition: Botany, Production and Uses. CAB International, Wallingford, UK, pp. 118. Murray, M.G., Thompson, W.F., 1980. Rapid isolation of high molecular weight plant DNA. Nucleic Acids Res. 8, 43214326. Nei, M., 1973. Analysis of gene diversity in subdivided populations. Proc. Natl. Acad. Sci. U. S. A. 70, 33213323. Nei, M., Li, W.H., 1979. Mathematical model for studying genetic variation in terms of restriction endonucleases. Proc. Natl. Acad. Sci. U. S. A. 76, 52695273. Paetkau, D., Calvert, W., Stirling, I., Strobeck, C., 1995. Microsatellite analysis of population structure in Canadian polar bears. Mol. Ecol. 4, 347354. Pritchard, J.K., Stephens, M., Donnelly, P., 2000. Inference of population structure using multilocus genotype data. Genetics 155, 945959. Pritchard, J.K., Wen, W., Falush, D., 2010. Documentation for Structure Software: Version 2.3. Chicago, http://pritch.bsd.uchicago.edu/structure software/ release versions/v2.3.4/structure doc.pdf Ravishankar, K.V., Anand, L., Dinesh, M.R., 2000. Assessment of genetic relatedness among mango cultivars of India using RAPD markers. J. Hortic. Sci. Biotechnol. 15, 198201. Ravishankar, K.V., Mani, B.H.R., Anand, L., Dinesh, M.R., 2011. Development of new microsatellite markers from mango (Mangifera indica) and cross-species amplication. Am. J. Bot. 98, e96e99. Samal, K.C., Jena, R.C., Swain, S.S., Das, B.K., Chand, P.K., 2012. Evaluation of genetic diversity among commercial cultivars, hybrids and local mango (Mangifera indica L.) genotypes of India using cumulative RAPD and ISSR markers. Euphytica 185, 195213. Samavi, H., Saiedi, G.H., 1991. Identication and collection of mango genotypes in Hormozgan province. Annual Report of Hormozgan Center of Seed and Seedling Investigation and Breeding, p. 12. Schnell, R.J., Ronning, C.M., Knight, R.J., 1995. Identication of cultivars and validation of genetic relationships in Mangifera indica using RAPD markers. Theor. Appl. Genet. 90, 269274. Schnell, R.J., Olano, C.T., Quintanilla, W.E., Meerow, A.W., 2005. Isolation and characterization of 15 microsatellite loci from mango (Mangifera indica L.) and cross-species amplication in closely related taxa. Mol. Ecol. Notes 5, 625627. Shamili, M., Talaie, A., Fatahi, R., Talebi, M., 2011. Morphological evaluation of some mango genotypes in Iran. Iran. J. Hortic. Sci. 41, 95110. Viruel, M.A., Escribano, P., Barbieri, M., Ferri, M., Hormaza, J.I., 2005. Fingerprinting, embryo type and geographic differentiation in mango (Mangifera indica L.) Anacardiaceae) with microsatellites. Mol. Breed. 15, 383393. Wright, S., 1951. The genetical structure of populations. Ann. Eugen. 15, 323354. Wnsch, A., Hormaza, J.I., 2002. Cultivar identication and genetic ngerprinting of temperate fruit tree species using DNA markers. Euphytica 125, 5967. Yeh, F.C., Young, R.C., Timothy, B., Boyle, T.B.J., Ye, Z.H.,1997. Popgene, the UserFriendly Shareware for Population Genetic Analysis. Mol. Biol. Biotech. Center, University of Alberta, Canada.

References
Adato, A., Sharon, D., Lavi, U., Hillel, J., Gazit, S., 1995. Application of DNA ngerprints for identication and genetic analysis of mango (Mangifera indica L.) genotypes. J. Am. Soc. Hortic. Sci 120, 259264. Bompard, J.M., 2009. Taxonomy and systematics. In: Litz, R.E. (Ed.), The Mango, 2nd Edition: Botany, Production and Uses. CAB International, Wallingford, UK, pp. 1941. Chiang, Y.C., Tsai, C.M., Chen, Y.K., Lee, S.R., Chen, C.H., Lin, Y.S., Tsai, C.C., 2012. Development and characterization of 20 new polymorphic microsatellite markers from Mangifera indica (Anacardiaceae). Am. J. Bot., e117e119. Degani, C., El-Batsri, R., Gazit, S., 1990. Enzyme polymorphism in mango. J. Am. Soc. Hortic. Sci. 115, 844847. Duval, M.F., Bunel, J., Sitbon, C., Risterucc, A.M., 2005. Development of microsatellite markers for mango (Mangifera indica L.). Mol. Ecol. Notes 5, 824826. Eiadthong, W., Yonemori, K., Sugiura, A., Utsunomiya, N., Subahadrandhu, S., 1999. Identication of mango cultivars of Thailand and evaluation of their genetic variation using the amplied fragments by simple sequence repeat- (SSR-) anchored primers. Sci. Hortic. 82, 5766. Eiadthong, W., Yonemori, K., Kanzaki, S., Sugiura, A., 2000. Amplied fragment length polymorphism analysis for studying genetic relationship among Mangifera species in Thailand. J. Am. Soc. Hortic. Sci. 125, 160164. Evans, E.A., Mendoza, O.J., 2009. World mango trade and the economics of mango production. In: Litz, R.E. (Ed.), The Mango, 2nd Edition: Botany, Production and Uses. CAB International, Wallingford, UK, pp. 606627. Excofer, L., Laval, G., Schneider, S., 2005. Arlequin ver. 3.0: an integrated software package for population genetics data analysis. Evol. Bioinform. Online 1, 4750. Falush, D., Stephens, M., Pritchard, J.K., 2003. Inference of population structure using multilocus genotype data: linked loci and correlated allele frequencies. Genetics 164, 15671587. Falush, D., Stephens, M., Pritchard, J.K., 2007. Inference of population structure using multilocus genotype data: dominant markers and null alleles. Mol. Ecol. Notes 7, 574578. FAOSTAT, 2012. FAO Statistics Database on the World Wide Web. http://faostat. fao.org/site/567/default.aspx#ancor (accessed August 2012). Hemanth Kumar, N.V., Narayaswamy, P., Theertha Prasad, D., Mukunda, G.K., Sondur, S.N., 2001. Estimation of genetic diversity of commercial mango cultivars using RAPD markers. J. Hortic. Sci. Biotechnol. 76, 529533. Hirano, R., Htun-Oo, T., Watanabe, K.N., 2010. Myanmar mango landraces reveal genetic uniqueness over common cultivars from Florida, India, and Southeast Asia. Genome 53, 321330. Honsho, C., Nishiyama, K., Eiadthong, W.K., 2005. Isolation and characterization of new microsatellite markers in mango (Mangifera indica L.). Mol. Ecol. Notes 5, 152154. Hubisz, M.J., Falush, D., Stephens, M., Pritchard, J.K., 2009. Inferring weak population structure with the assistance of sample group information. Mol. Ecol. Resour. 9, 13221332. IPGRI, 2006. Descriptors for Mango (Mangifera indica L.). International Plant Genetic Resources Institute, Rome, Italy. Iyer, C.P.A., Degani, C., 1997. Classical breeding and genetics. In: Litz, R.E. (Ed.), The Mango-Botany, Production and Uses. CAB International, Wallingford Oxon, pp. 4968. Karihaloo, J.L., Dwivedi, Y.K., Archak, S., Gaikwad, A.B., 2003. Analysis of genetic diversity of Indian mango cultivars using RAPD markers. J. Hortic. Sci. Biotechnol. 78, 285289. Kashkush, K., Jinggui, F., Tomer, E., Lavi, U., 2001. Cultivar identication and genetic map of mango (Mangifera indica L.). Euphytica 122, 129136.