Loading...
Thesis
Browse
16 results
Search Results
ThesisItem Open Access MOLECULAR SCREENING OF ALPHONSO MOTHER PLANT (Mangifera indica L.) AND ITS SEEDLINGS THROUGH MOLECULAR MARKERS(Dr.Balasaheb Sawant Konkan Krishi Vidyapeeth, Dapoli, 2023-02-06) CHOUKSEY, MAHENDRA KUMAR; BEDSE, T. J.; SAWARDEKAR, S. V.; DESHPANDE, R.S.; DALVI, V.V.; Mahadik, S. G.The most valued table cultivar of mango (Mangifera indica L.), known as "Alphonso," has been growth in India for more than a century. In order to explore intracultivar heterogeneity based on microsatellite markers, 15 seedlings of Alphonso mango were grown by the stones taken from identified mother plant for Agriculture Research station, Shirgaon, Dist.Ratnagiri (Maharashtra). It was investigated that whether there was genetic heterogeneity or relatedness between the chosen seedlings and their mother plant. The all fourteen verified mango-specific simple sequence repeats were found polymorphic. In the result we found total 224 alleles were produced by polymorphic microsatellites, they showed the 100% polymorphism.ThesisItem Open Access STANDARDIZATION OF IN VITRO ORGANOGENESIS TECHNIQUE IN BAMBOO(Dr.Balasaheb Sawant Konkan Krishi Vidyapeeth, Dapoli, 2023-02-03) CHAVHAN, SAMIKSHA RAMESHWAR; SAWARDEKAR, S.V.; DESHPANDE, R.S.; PETHE, U.B.; DALVI, V.V.ThesisItem Open Access STUDY OF IN VITRO REGENERATION TECHNIQUE THROUGH FLORAL ORGAN OF BANANA, CV. GRAND NAINE (G-9)(Dr.Balasaheb Sawant Konkan Krishi Vidyapeeth, Dapoli, 2023-02-07) PAWDE, GOVIND BHAURAO; SAWARDEKAR, S. V.; Deshpande, R.S.; Kunkerkar, R.L.; Pethe, U.B.ThesisItem Open Access SCREENING OF RICE (Oryza sativa L.) CULTURES FOR BIOTIC AND ABIOTIC STRESS USING SSR MARKERS.(Dr.Balasaheb Sawant Konkan Krishi Vidyapeeth, Dapoli, 2023-01-31) SAWANT, HEMANT SANTOSH; DESHPANDE, R. S.; Sawardekar, S.V.; Mahadik, S.G.; Kunkerkar, R.L.ThesisItem Open Access STANDARDIZATION OF In Vitro CULTURE ESTABLISHMENT TECHNIQUE IN MANGO (Mangifera Indica L.)(Dr. Balasaheb Sawant Konkan Krishi Vidyapeeth, Dapoli., 2021-06) KARANJE, PRANIT DHANANJAY; SAWARDEKAR, S.V.; DESHPANDE, R.S.; Parulekar, Y. R.The research was aimed to standardize the sterilization technique and to optimize the in vitro culture establishment in mango using apical meristem, axillary bud and leaf of three different varieties of mango viz. Alphonso, Kesar and Ratna. The maximum aseptic culture was in treatment T8 (86.81%) consisting Wiping with Ethanol + Running Water + Tween 20 + Dettol + Savlon + Polyvinylpyrrolidone + Activated Charcoal + Carbendazim + Mercuric Chloride + Cefotaxime. Similarly, media combination M9 consisting WPM + 5 mgL-1 BAP + 0.5 mgL-1 NAA was found best as it recorded 84.04% culture establishment.ThesisItem Open Access “STUDY OF IN VITRO REGENERATION TECHNIQUE IN ELEPHANT FOOT YAM (Amorphophallus paeoniifolius L.)” T06802(Dr. Balasaheb Sawant Konkan Krishi Vidyapeeth, Dapoli, 2020-10) PAWAR, PRAVIN BHAUSAHEB; Sawardekar, S. V.The present investigation entitled, “Study of in vitro regeneration technique in elephant foot yam (Amorphophallus paeoniifolius L.)” was aimed to standardize surface sterilization technique and to standardize culture establishment technique for in vitro regeneration of elephant foot yam. The study was conducted in completely randomized design with 3 replications. The surface sterilization treatment of 750 mg/L Carbendazim (overnight) before cut, 12.5 ml/L Sodium hypochlorite (20 min), 10 ml/L Tween 20 (15 min), 10 ml/L Dettol and 50 ml/L Savlon (30 min), 1 g/L Carbendazim (10 min), under running warm water (1 hr), 2 g/L Carbendazim (15 min), 70 % Ethanol (2 min), 0.12 g/L Mercuric Chloride (15 min), 50 ml/L Sodium hypochlorite (5 min), 16 ml/L GA3 (10 min), + 3 ml/L Streptomycin (20 min), 750 mg/L Cefotaxime (I) (20 min) and 750 mg/L Cefotaxime (II) (40 min) was found to be best combination to achieve highest percentage of aseptic cultures (88.57%). Corm bud explants transferred to MS medium containing different combinations of plant growth regulators for culture establishment. Among the various combinations, MS medium supplemented with 6 mg/LBAP + 0.5 mg/L NAA showed highest percentage of culture establishment (60%). The media combination of MS + 6 mg/L BAP + 0.5 mg/L NAA + 0.1 g/L Ascorbic acid was found effective for highest percentage (88.89%) of shoot induction. The number of days required for first shoot initiation ranged from 15.33 to 23.37 days. The media combination comprised of MS + 6 mg/L BAP + 0.5 mg/L NAA + 0.1 g/L Ascorbic acid showed minimum (15.33) days for shoot induction. Same media combination was also found effective for highest average number of multiple shoot bud induction (5.33). However, all the hormonal combinations failed to induce in vitro rooting. Key words: Elephant foot yam, surface sterilization, culture establishment, shooting media.ThesisItem Open Access MOLECULAR PROFILING OF MANGO (Mangifera indica L.) HYBRIDS AND THEIR PARENTS USING MOLECULAR MARKERS(Dr. BSKKV., MH, 2020-10) BHOSALE, POOJA SUNIL; Deshpande, R.S.The present study was carried out with an objective to study the band sharing pattern in mango (Mangifera indica L.) hybrids and their parents using molecular markers and to analyse the similarity and distinctiveness between them. For the present study, the hybrids developed by Dr. Balasaheb Sawant Konkan Krishi Vidyapeeth, Dapoli were used. These included Ratna, Sindhu, Konkan Ruchi, Suvarna and Konkan Samrat. The parent varieties comprised of Alphonso, Neelum, Ratna and Tommy Atkins. DNA was extracted from tender leaf samples collected from Regional Fruit Research Station, Vengurle. Using the rapid method, the five components of the extraction buffer were standardized. The results obtained using 0.900 g glucose, 0.100 g PVP, 0.040 g sodium bisulphite, 0.050 g sodium lauryl sulphate, 500 µl sarcosyl were most suitable. Modification in PCR parameters like PCR master mixture and thermo profile resulted in clear and specific banding pattern. The average per cent polymorphism showed by 13 ISSR primers amongst four parent varieties and five hybrid varieties was 75.64 & 79.41 per cent, respectively. The average per cent polymorphism across the 13 primers amongst eight mango varieties was 83.92 per cent. The polymorphic information content (PIC) values ranged from 0.575 to 0.913 with an average PIC value of 0.825 per primer. UPGMA followed the similar pattern of grouping the individuals; i.e. it separated all the varieties into two main clusters. The results of present study indicated the efficiency of ISSR markers in examining the genetic variation in the crop at molecular level. The study helps in getting the molecular profile of each hybrid which will assist the mango growers to identify the planting material at the juvenile stage itself. The findings of this molecular study can establish the identity and creditability of the variety at molecular level and substantiate its claim in PPVFRA registration at national and international level. Keywords: Mango, Hybrids, Banding pattern, Polymorphism,ISSR, Marker.ThesisItem Open Access DNA BARCODING FOR IDENTIFICATION OF RICE (Oryza sativa L.) VARIETIES DEVELOPED BY Dr. BALASAHEB SAWANT KONKAN KRISHI VIDYAPEETH, DAPOLI (Accession No. T06800)(Dr. Balasaheb Sawant Konkan Krishi Vidyapeeth, Dapoli, 2020-10) MHATRE, DHANASHREE PRASHANT; Sawardekar, S.V.The present study was conducted with an objective to characterize and add a molecular tag i.e. the DNA barcode for the 20 varieties developed by Dr. Balasaheb Sawant Konkan Krishi Vidyapeeth, Dapoli. The study involved analysis of these varieties by using 10 SSR markers and 10 barcoding loci markers. The evolutionary analysis and phylogenetic relationship study was done using MEGA software. CLUSTAL Omega online tool detected the conserved region in the varieties. DNA barcodes derived from the chloroplast genome were used to identify varieties and in the conservation of breeding resources. The gene regions are chosen because they have less intraspecific (within species) variations than interspecific (between species) variations, which are known as the “Barcoding Gap”. DNA barcoding has wider applications in the fields of taxonomy, conservation and identifications of different crops, trees and plants. Two major types of seed grains were observed as bold and slender and categorized as short and long studied on the basis of morphological data. Amplification efficiency observed was about 100 % in 3 primers to 81.25 % in remaining primers. Fragment size range was 600-2900 bp in 10 loci. Sequencing efficiency of loci ranged from 100% to 18.75%. The estimated evolutionary divergence between sequences ranged from 0.000-2.170 (average 0.001-0.405). The Maximum Likelihood values of maximum transitional and transversional rate were observed. The parsimony informative sites was estimated with maximum 161 sites recorded in psbA-trnH, followed by 159 sites trnK and 33 rbcL, 21 in matK and number of variable sites reported highest in psbA-trnH 568, trnK 457 and matK-1M- matK3RIM 285.While nucleotide diversity (per site pi) reported maximum in trnK 0.1455. Phylogenetic relationship established using Neighbor Joining Method for all the 10 loci distinctly separated out Ratnagiri-2 for matK, Palghar-2 for psbA-trnH , Karjat-1 And Karjat-2 for trnH-psbA , Ratnagiri-4 for atpH-atpI , Panvel-2 for petA- psbJ , Karjat-7 for trnK and Ratnagiri-7 for matK-1M-matK3RIM . Both strand of psbA-trnH and trnH-psbA showed good discrimation power, also petA-psbJ showed higher discrimination. trnK distinctly separated four of the 16 varieties in alignments. rbcL did not show discrimination but more of conserved region was seen whereas matK could give an average discrimination among the varieties. Barcodes were generated using online tool for the different loci considering the variations in sequence at nucleotide level. Comparative analysis of SSR markers was done with RM 338 highly informative with PIC value of 0.631 and observing the most frequent band size among 20 varieties for the microsatellites markers. Jaccard’s similarity coefficient ranged from 0.190 to 0.750. Cluster analysis was established by UPGMA and similarity index by Jaccard’s coefficient. In conclusion, this study provides a precursive assessment data that will be useful for extensive application of DNA Barcoding in not only rice varieties but other fruit crops, ornamental and forestal plants of Dr. Balasaheb Sawant Konkan Krishi Vidyapeeth, Dapoli. Keywords: DNA barcoding, matK, psbA-trnH, trnH-psbA, rbcL, trnK, Parsimony informative sites, Neighbor Joining Method, CLUSTAL Omega, MEGA 7.0.ThesisItem Open Access STANDARDIZATION OF SEX DISCRIMINATION TECHNIQUE IN KOKUM (Garcinia indicaChoisy) USING MOLECULAR MARKERS(Dr. Balasaheb Sawant Konkan Krishi Vidyapeeth, Dapoli, 2020-10) GAJARE, TUSHAR POPAT; Deshpande, .R.S.In the present investigation it was attempted to standardize the sex discrimination technique in Kokum by considering hermaphrodites along with male and female by using ISSR markers. The study utilized five samples each of male, female(cv.Konkan Amruta)and hermaphrodite of Kokum which were identified on the basis of their flowering and fruiting behavior. DNA isolated using rapid method was used for PCR amplification using 60 ISSR primers. Out of the 60 primers used for the study, 29 primers did not amplify, 3 primers showed intangible amplification, 23 primers were monomorphic and five were polymorphic. Out of these five polymorphic primers, UBC 807(Male specific), UBC 844(Hermaphrodite specific) and UBC 845(Female specific) were not reproducible. Only two primers namely, UBC 825 and UBC 878 were reproducible with polymorphism of 20 and 63.63 percent respectively. The polymorphic bands UBC 8252000,2100(Female specific), UBC 8781600(female specific), UBC 878780,980 (hermaphrodite specific) were faint but reproducible.UBC 878 also produced two bands, UBC 878990,1900in female and hermaphrodite but absent in males, three bands UBC 8781480,UBC 8781000 and 980 only in male and females and absent in hermaphrodites. The similarity matrix and cluster analysis revealed that hermaphrodites are closely related to males than the females. Thus these markers successfully standardized the sex discrimination technique in Kokum. They can further be utilized for SCAR development and understanding the evolution of dioeciousness and various pathways and processes associated with it. Keywords: Garcinia indicaChoisy, sex discrimination, ISSR, polymorphism.
