STUDIES ON GENETIC CHARACTERIZATION OF PERSIAN WALNUT (Juglans regia L.) GERMPLASM USING MOLECULAR MARKERS
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Date
2018-02
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UHF,NAUNI
Abstract
ABSTRACTJuglans regia L
. commonly known as ‘Akhrot’ and it belongs to family Juglandaceae.Walnut oil contains alphalinolenic
acid and linoleic acid.They reduces the risk of cardiovascular diseases. In the present study EST-SSR
markers were datamined for Juglans regia L.and were used for polymorphism studies. EST sequences of
Juglans regia L.were downloaded from NCBI website (www.ncbi.nlm.nih.gov/nucest). 2000 EST sequences for
Juglans regia L. were screened. ESTs containing SSR motifs were extracted out using an online tool, SSRIT
(www.gramene.org/db/searches/SSRtool). SSR primers were designed from the EST-SSR containing sequences
using PRIMER 3 software (www.frodo.wi-mit.edu/primer3/) and 98 primers were custom synthesized.
Polymorphism study was carried out using 35 EST-SSR, 29 Genomic SSR and 30 ISSR primers among 37
genotypes of Juglans regia L. For polymorphism, DNA was isolated from young leaves of all the 37 genotypes
using CTAB method (Doyle and Doyle, 1987). 24 primers were informative out of 35 EST-SSR, 19 primers
were informative out of 29 Genomic SSR and 22 primers were informative out of 30 ISSR. Jaccard’s similarity
matrix was developed and dendrograms were generated using NTSYSpc ver.2.02h to establish the percent
similarity among the 37 genotypes of Juglans regia L. With EST-SSRs, two clusters ‘A’ and ‘B’ were obtained.
Cluster A was further subdivided into A1 and A2. A1 contained 28 genotypes and A2 contained eight genotypes
whereas cluster B contained only one genotype. For Genomic SSR two clusters ‘C’ and ‘D’ were obtained.
Cluster C was further subdivided into C1 and C2. C1 contained 33 genotypes and C2 contained three genotypes
whereas cluster D contained only one genotype whereas For ISSRs, 37 genotypes were divided into two cluster
i.e ‘E’ and ‘F’. Cluster E was further subdivided into E1 and E2. E1 contained 28 genotypes and E2 contained
eight genotypes whereas cluster F contained only one genotype. Morpho-physical characters were also studied
for diversity study like Time of Bud burst, Flowering time, Degree of Dichogamy, Nut shape, Shell hardness,
Nut weight, Kernel weight, Kernel recovery.Eucledia statistical tool is used to calculate the dissimilarity by
combining both molecular as well as morphological data.Thus molecular markers and morpho-physical
charaters used in the present study revealed a high level of polymorphism in the 37 genotypes of Juglans regia
L., revealing their efficiency for diversity analysis studies.
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