Morphological and Molecular Characterization of Maize inbreed Lines (Zea mays L.)

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dc.contributor.advisorKumar, Ajay
dc.contributor.authorKumar, Shashi
dc.date.accessioned2018-02-07T10:49:01Z
dc.date.available2018-02-07T10:49:01Z
dc.date.issued2017
dc.description.abstractA study was undertaken to characterize and evaluate eighteen locally adapted inbred lines of maize for various morphological characters, to investigate the SSR marker based polymorphism for characterization and differentiation of these maize inbred lines by using polymorphic and informative SSR markers and to estimate the extent of genetic diversity among these maize inbred lines using morphological and molecular characterization. The present investigation field research work was undertaken during kharif 2015 in Randomized Block Design (RBD) with 3 replication having plot size of 1.5 x 4.0 = 6 m2 at TCA Dholi farm, Dr.R.P.C.A.U., Pusa, Samastipur, (Bihar) and molecular work was performed in the molecular breeding laboratory, Department of Plant Breeding and Genetics, Dr. R.P.C.A.U., during kharif 2015. The characters studied were Days to 75% tasseling, Days to 75% silking, Days to 75% brown husk, Plant height(cm), Ear height(cm), Ear length(cm), Ear girth (cm), Number of kernel rows per ear, Number of kernels per row and Grain yield (Kg/ha). The Inbred lines were grown at TCA, Dholi Research Station farm for extraction of genomic DNA from the young seedlings. The morphological characterization include biometrical analysis namely, genetic variability, heritability (broad sense), genetic advance as per cent of mean, Character association analysis, Path analysis, genetic divergence using D2 statistic by Tocher’s method Similarly Molecular characterization of maize inbred lines involved: Extraction of genomic DNA, Quantification of genomic DNA, Amplification of genomic DNA, Separation of amplified DNA, Statistical analysis of molecular markers based data, Assessment of divergence based on molecular markers Scoring of bands using Quantity 1D analysis software of Biorad Inc., USA and Preparation of dendogram using NTSYS-pc version 2.0 software (Rohlf, 2000) with thirteen SSR Primers. Analysis of variance revealed highly significant differences among the inbred lines for all the parameters except Days to 75% tasseling, Days to 75% silking, Days to 75% brown husk and Ear girth. From the variability studies, high magnitude of GCV and was observed for Grain yield (Kg/ha) indicating the importance of these traits in evolution and selection of inbred lines for yield improvement under maize breeding programme. High heritability accompanied with high genetic advance as percent of mean was recorded for ear girth, Number of kernels per row and grain yield (Kg/ha) indicating effectiveness of selection due to preponderance of additive gene action for these traits and direct selection may be effective for these traits. Correlation studies indicated that Days to 75% brown husk, Plant height (cm), Ear height (cm), Ear length (cm), Ear girth (cm), Number of Kernel rows/ear, number of kernels/row showed significant positive association with grain yield as well as among themselves at phenotypic and genotypic level. Highest positive direct effect was exhibited by Days to 75% brown husk, Ear height (cm), Ear length (cm), Ear girth (cm),Number of Kernel rows/ear and Grain yield (Kg/ha). Hence, selection based on these characters would be more effective for yield improvement. All the 18 inbred lines were grouped into 5 clusters using D2 statistics using Tochers method (Morphological characterization) and 8 clusters using Jaccard similarity coefficient analysis (Molecular characterization) shows that inbred lines viz., PoP-61 and LM-13 (cluster I, cluster A), WNC DMR 11 R 27290 (cluster IV, cluster D) and S99 TLYQ(HG-AB)-BBB-54-BBB-54-BBB/CML-193 (cluster V, cluster E) included in the same cluster indicating thereby that these inbred lines of maize are relatively more closely related to each other with respect to the morphological and molecular characterization under consideration in the present investigation. The maximum intra cluster distance was observed in cluster V and minimum intra cluster distance was observed in cluster I. The highest inter cluster distance was observed in between Cluster I and V. Genotype from these clusters may be selected as parents for hybridization programme for developing new hybrid combinations. Among 153 combinations, number of kernels per row ranked 1st (93 time) and maximum in Percent contribution towards total divergence, Cluster mean in cluster I (51.00, 15.72, 13.11, 11.56, 1400.75) but it showed maximum in cluster V (110.67, 72.83, 19.00, 15.00, 14.00, 3010.56) respectively.Therefore, selection of parents from different clusters might be done for desired traits on the basis of higher cluster mean values, suggesting scope for Improvement in these characters. A panel of thirteen primers, namely, phi083, nc133, phi029, phi053, umc1304, umc1332, phi034, umc1161, phi014, phi065, umc1367, umc1196 and umc1266 generated amplified products due to amplification of two locus and targeting each of the chromosome no. 2, 2, 3, 3, 8, 5, 7, 8, 8, 9, 10, 10 and 3 chromosomes of the maize genome was successfully used to achieve the targeted amplification and the polymorphism was recognized on the basis of presence or absence of bands, in addition to variation in respect of number and position of bands from seven in the cases of nc133, umc1161, umc1367 and umc1196 to ten in the cases of, phi029, umc1332, phi034 and umc1266. Altogether 110 alleles (62 shared and 48 unique allelic variants) were detected among the eighteen maize inbred lines with an average of 8.46 alleles per locus using 13 SSR markers. The number of unique alleles per locus ranged from two in the case of nc133 to six alleles in the case of umc1266. Umc1266, phi029, phi034, umc1304, umc1332, umc1196, phi083, umc1161, phi014, phi065, umc1367 and nc133 generated considerably greater percentage of unique alleles in descending order of magnitude. The SSR locus associated with primer pairs phi083, nc133, phi053, umc1161, phi014, umc1367and umc1196 showed null alleles in some of the inbred lines under evaluation reflecting the failure of locus specific SSR primer directed generation of amplified products. In general, the SSR loci with di-nucleotide repeat motifs tended to detect greater number of alleles than the locus with tri-nucleotide, teta-nucleotide or penta-nucleotide repeat sequence. The marker with a GA or AG repeat motif showed greater variability than marker with a CT or TG or TA repeat motif among the loci with perfect di-nucleotide repeat motifs. Presence of stutter bands were observed in the case of tri and tetra-nucleotide repeat sequences detected by primer pairs umc1304 and umc1332. The PIC values revealing allele diversity and frequency among the inbred lines varied from 0.805 in the case of umc1367 to 0.886 in the case of phi065 with an average of 0.849 per primer. Since PIC values higher than 0.5 reflect high informative ability of the markers, all the primer pairs with thewere highly informative. The pair-wise estimates of similarity coefficients ranged from 0.000 to 0.375. The magnitude of similarity coefficient between Pool 33- 193 and Pool 34- 193 (0.375) was the maximum amongst pair-wise combinations of entries under evaluation in the present study. By drawing the phenon line at 0.11 similarity units. A perusal of the dendrogram clearly indicates that the entries were basically divided into three groups. The first multi-genotypic group consisted consisted of ten inbred lines, second di-genoptypic group consisted of six inbred lines and third mono-genotypic group consisted of two inbred line. Use of SSR markers appeared more efficient in achieving unique and unambiguous characterization and differentiation of inbred lines used in the present study. The SSR analysis also revealed unique or variety specific allele, which could be useful as DNA fingerprints in the identification and preservation of maize inbred lines.en_US
dc.identifier.urihttp://krishikosh.egranth.ac.in/handle/1/5810040570
dc.keywordsMaize, Zea mays, Molecular characterization, Morphological characterization,en_US
dc.language.isoenen_US
dc.pages137+viiien_US
dc.publisherDr. Rajendra Prasad Central Agricultural University, Pusa (Samastipur)en_US
dc.subGenetics and Plant Breedingen_US
dc.subjectnullen_US
dc.themeTo characterize and evaluate eighteen locally adapted inbred lines of maizeen_US
dc.these.typeM.Scen_US
dc.titleMorphological and Molecular Characterization of Maize inbreed Lines (Zea mays L.)en_US
dc.typeThesisen_US
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