Dr. Panjabrao Deshmukh Krishi Vidyapeeth, Akola
Dr. Panjabrao Deshmukh Krishi Vidyapeeth, Akola was established on 20th October, 1969 with its head-quarter at Akola. This Agricultural University was named after the illustrious son of Vidarbha Dr. Panjabrao (alias Bhausaheb) Deshmukh, who was the Minister for Agriculture,Govt. of India. The jurisdiction of this university is spread over the eleven districts of Vidarbha. According to the University Act 1983 (of the Government of Maharashtra), the University is entrusted with the responsibility of agricultural education, research and extension education alongwith breeder and foundation seed programme.
The University has its main campus at Akola. The instructional programmes at main campus are spread over in 5 Colleges namely, College of Agriculture, College of Agricultural Engineering & Technology, College of Forestry, College of Horticulture and Post Graduate Institute. At this campus 4 degree programmes namely B.Sc.(Agri.) B.Sc. (Hort.), B.Sc. (Forestry) and B.Tech. (Ag. Engg.) , two Master’s Degree Programmes viz. M.Sc.(Agri.) and M.Tech. (Agri.Engg.) and Doctoral Degree Programmes in the faculties of Agriculture and Agril. Engineering are offered.
The University has its sub-campus at Nagpur with constituent College, College of Agriculture which offers B.Sc.(Agri.) and M.Sc.(Agri.) degree programmes. The Nagpur Campus is accomplished with a garden, surrounded by its natural beauty and a well established Zoo which attract the general public and visitors to the city. A separate botanic Garden is being maintained on 22 hectares with a green house for the benefit of research workers.
In addition there are 2 affiliated grant-in-aid colleges and 14 private non-grant-in-aid colleges under the umbrella of this University
A Central Research Station is situated at the main Campus which caters to the need of research projects undertaken by Crop Scientists of the principle crops of the region are Cotton, Sorghum, Oilseeds and Pulses.
Browse
Browsing Dr. Panjabrao Deshmukh Krishi Vidyapeeth, Akola by Subject "Agricultural Biotechnology"
Now showing 1 - 18 of 37
Results Per Page
Sort Options
ThesisItem Open Access ANALYSIS OF DNA METHYLATION PATTERN ASSOCIATED WITH FLORAL BUD DISTORTION IN SOYBEAN (Gylcine max L.).(Dr. Panjabrao Deshmukh Krishi Vidyapeeth, Akola, Maharashtra., 2017-08-28) KAD, MISS. SNEHAL KRISHNA; Jadhav, Dr. P. V.The present investigation entitled, “Analysis of DNA methylation pattern associated with floral bud distortion in soybean (Glycine max L.)” was carried out during 2016-17 at Biotechnology Centre, Department of Agricultural Botany, Dr. Panjabrao Deshmukh Krishi Vidyapeeth, Akola to investigate methylation pattern associated with floral bud distortion in soybean. DNA methylation is known to play an important role in regulating gene expression during biological development and tissue differentiation in eukaryotes. In the present attempt, cytosine methylation profile was determined in elite soybean (Glycine max L.) cultivar JS-335. Methylation-sensitive amplified polymorphism was used to generate genome-wide methylation profiles and compared the patterns of cytosine methylation in floral bud distorted and asymptomatic plant. It has been learned that soybean plant is affected by the disorder known as floral bud distortion (FBD), no pod formation take place at R6 stage which results in huge yield loss in symptomatic plant(s). The methylation-sensitive amplified polymorphism technique (MSAP) which is a modification of amplified fragment length polymorphism (AFLP) used to investigate the epigenetic changes associated with FBD in soybean. The samples of genomic DNA were double-digested with either HpaII or MspI and with EcoRI. To analyze epigenetic changes, 25 MSAP primer combinations detected a few DNA methylation patterns in the plant tissue of symptomatic and asymptomatic plant. A total of 378 and 392 clear and reproducible bands were amplified from symptomatic and asymptomatic samples, respectively. In the symptomatic plant, out of 378 MSAP sites, 273 (72.22%) were found un-methylated, 31 (8.20%) were hemi-methylated, 35 (9.25%) were fully methylated; and 39 (10.31%) hyper-methylated sites were amplified. Whereas, in asymptomatic plant, out of 392 MSAP sites, 315 (80.35%) found un-methylated, 21(5.35%) hemi-methylated, 29 (7.39%) fully methylated and 25 (60.37) hyper-methylated sites were amplified. An increased level of methylation was recorded in symptomatic plant (27.28%) than that of asymptomatic (19.13%). In fact, there was evidence of increase in fully methylated ratio to 19.57% in symptomatic plant and that in case of asymptomatic revealed decrease of 12.75%. These results indicated that there were greater differences of the genomic DNA methylation extent and also variation in the pattern of methylation between the symptomatic and asymptomatic plant. It revealed that the methylation status varied substantially during the development stage of flower. The polymorphic DNA methylation fragments that showed polymorphic bands were excised from Polyacrylamide gel and cloned in vector pTZ57R/T. However, total 10 polymorphic bands were excised varying from 150 to 250 bp, out of which only 2 bands were successfully purified from the PAGE. Further these methylated fragments were cloned in vector pTZ57R/T (2886 bp). These findings successfully generated extent and pattern of cytosine methylation in floral bud distorted and asymptomatic plants. In the present investigation, although limited numbers of methylation sensitive polymorphic fragments were obtained this helped to calculate the extent and pattern of methylation associated with floral bud distortion in soybean.ThesisItem Open Access BIOCHEMICAL CHARACTERIZATION OF WHEAT LINES UNDER WATER STRESS CONDITION(Dr. Panjabrao Deshmukh Krishi Vidyapeeth, Akola, Maharashtra., 2022-01-11) KHARADE, SHUTIJA, JAYDEEP; Gahukar, Dr. S. J.Drought is multidimensional in nature affecting plant at different level of organization. Under drought conditions, crops close their stomata to limit evaporative water loss, thus reducing carbon uptake by photosynthesis and decreasing yields. Plants sustain water deficit by drought escape, drought avoidance or drought tolerance which all contribute to resistance. Plants endure drought through osmotic alteration, stomatal resistance, improved water uptake by making changes in their metabolism. These traits can be useful in selecting drought tolerant cultivars for water stressed areas. The present study was planned assaying water stress tolerance in wheat genotypes. In this study various morphological, physiological and biochemical observations were recorded. An in vitro study was conducted in which drought stress was artificially created using various concentrations of PEG-6000 (0 %, 10 % and 15%) in a completely randomized block design. Various physiological parameters were recorded. Similarly, an in vivo study was conducted on field where wheat genotypes were grown at different irrigation levels, in control (six irrigation) and water stressed (three irrigation) condition in a randomized block design. These were then evaluated for various physiological, morphological and biochemical parameters. In the in vitro study, the physiological studies showed that germination percentage, shoot-root length, seedling vigour index, relative water content and chlorophyll content decreased with increasing osmotic stress. Osmotic stress showed negative effect on all the studied parameters. In the in vivo study, analysis showed that all the morphological parameters i.e. plant height, tillers per plant, spike length, grains per spike, 1000 grain weight, yield per plant decreased in water stressed condition, while days to 50% flowering increased in water stressed condition. In physiological analysis, relative water content and chlorophyll content decreased in water stress condition. Similarly, in biochemical analysis it was observed that protein content has inverse relation with water stress while proline content and ascorbate peroxidase activity has direct relationship with increasing water stress. Genotypes AKAW-3717, AKAW-4842 and AKAW- 4924 were observed to be superior cultivars for low rainfall areas with possibility of drought spells as they performed well in both in vitro and in vivo studies. AKAW-2862-1 and AKAW-4925 showed a consistently poor performance in both the studies and found to be drought susceptible. It is thus concluded that water stress levels had extensive effect on germination and seedling growth. Morphological, biochemical, physiological analysis discovered that adequate difference for drought tolerance existed in the tested genotypes. AKAW-3717, AKAW-4842 and AKAW- 4924 may prove a promising parent material for breeding drought tolerant wheat cultivars and can be considered better genotypes for low rainfall or drought prone areas. Correlation study showed that relative water content and proline content has significant positive association with grain yield and thus these traits can be used as a selection criterion for such drought environment for further improvement.ThesisItem Open Access BIOLOGICAL PREPARATION OF NANOPARTICLES AND ITS STUDY ON MANAGEMENT OF POSTHARVEST DISEASES OF LEMON.(Dr. Panjabrao Deshmukh Krishi Vidyapeeth, Akola, Maharashtra., 2022-05-05) WAGHODE, KANCHAN HARIDAS.; GAHUKAR, Dr. S. J.The present investigation entitled “Biological preparation of nanoparticles and its study on management of post-harvest diseases of lemon” was conducted Food Microbiology Laboratory of All India Coordinated Research Project on Post–Harvest Engineering and Technology Dr. Panjabrao Deshmukh Krishi Vidyapeeth, Akola. Lemon, fruits are highly demanded in the market for its nutritional value. About 30% of fruits are affected or damaged by microorganisms, insect, pre and post harvesting conditions during transport and preservation. Coating is an effective and alternative method to solve these problems and reduce its post-harvest losses during storage condition. In this study utilization of silver nanoparticle, Aloe vera and its combination as a good coat up to 28 days and stored at different temperatures (cold condition and ambient condition) to reduce post-harvest losses. Nanoparticle is synthesized from fungus Penicillium digitatum and characterized by UV-Vis spectroscopy, Particle size analysis, Zeta potential and FTIR techniques. the UV-Visible spectra of the biosynthesized silver nanoparticle shows characteristic surface plasmon absorption at 379nm. The biosynthesized AgNPs were found to have negative zeta potential -10.3 mV which shows increases the stability of the formulation. Z-average of silver nanoparticle having 23.11 (r. nm) size and FTIR shows band at 1553, 1683, 2390, 3633, 3781, 3890 cm-1. The treatment combinations were used for coating are C1=control, C2=Aloe vera, C3= nanoparticle and C4= combination of Aloe vera and nanoparticle. The coatings as having a high potential to carry active ingredients such as nano-materials maintained rotting percent Average weight, total soluble solids, titratable acidity, ascorbic acid, pH, colour of fruits. All the treatments are found to be effective against rotting of fruits, reducing weight, ascorbic acid. Thus this study will help to reduce the post-harvest losses during storage. The minimum average weight was noted in uncoated fruits whereas maximum average weight was noted in fruits coated with combination of Aloe vera and nanoparticles. The minimum TSS, titrable acidity, total sugar, reducing sugar and pH were noted in cold storage condition while non reducing sugar, ascorbic acid and average weight were noted maximum in ambient condition storage.ThesisItem Open Access DECIPHERING THE DIFFERENTIALLY EXPRESSED GENES IN FOXTAIL MILLET (Setaria italica L.) IN RESPONSE TO WATER STRESS.(Dr. Panjabrao Deshmukh Krishi Vidyapeeth, Akola, Maharashtra., 2017-06-15) GAWAI, DIPTI CHANDRABHAN.; Gahukar, Dr. S. J.The present investigation entitled “Deciphering the differentially expressed genes in Foxtail millet (Setaria italica L.) in response to water stress” was carried out at the Biotechnology Centre, Department of Agricultural Botany, Dr. Panjabrao Deshmukh Krishi Vidyapeeth, Akola during 2013-2016. In the present study, sixty two accessions of foxtail millet (Setaria italica L.) were investigated for physiological, biochemical and molecular responses for water stress. Water stress was imposed under laboratory conditions by using PEG-6000 and by withholding water and performances of various genotypes were monitored against a control. Seedling traits such as germination percentage, root length, shoot length and seedling dry weight, various physiological parameters like relative water content (RWC), leaf water potential (LWP) and chlorophyll content (CC) were studied in unstressed and stressed experiments. The results showed that water deficit stress has exerted negative effect on all the physiological parameters considered. Based on these observations, most drought tolerant and susceptible accessions were selected. IC97087, IC97189, IC120159 and IC120239 were recorded as the most drought tolerant accessions, whereas, IC97109, IC120234, IC120346 and Lepakshi were recorded as susceptible accessions. The selected core set of accessions were then screened for their water stress tolerance on the basis of some primary metabolites like proline, total carbohydrates, starch and antioxidant enzymes like superoxide dismutase, peroxidase, catalase and glutathione reductase. Osmolytes viz. proline and carbohydrates increased with increasing drought stress. The proline content in unstressed tolerant plant is found to be much higher as compared to that in unstressed susceptible plants suggesting its important role in drought tolerance of plants. Photosynthetic pigment decreased with increasing drought stress. Activities of antioxidant enzymes increased with drought stress in most of the accessions. The activity of SOD was found to be highest in IC120239 which was found to be a tolerant accession in prior physiological screening. The activity of GR was high in all tolerant accession than in susceptible accessions. Catalase and POD activity was found to be highest in Prasad. Drought stress preferentially enhanced the activities of enzymatic antioxidants and accumulation of osmolytes. Further, gene expression studies were carried out using the two contrasting accessions IC97189 (tolerant) and IC97109 (susceptible), for deciphering the transcriptional regulation of abiotic stress-related genes. cDNA SRAP analysis showed a relatively higher number of genes expressed in IC97189. The differential gene expression of drought responsible transcriptional factors like DREB1, DREB2, Aquaporins and C2H2 was studied in selected drought tolerant and susceptible plants each one showing considerable difference gene expression. Genome–wide investigation of plant aquaporin protein from foxtail millet was carried out to show that aquaporins comprise of MIP superfamily with conserved motif. Expression profiling of PIPs showed differences in gene expression pattern. Further an attempt was made to characterise full length sequence of PIP 2;7 to isolate a 770bp long DNA stretch out of the 857 base pair CDS having high homology to a UNPRIDICTED PIP2;7 gene in Setaria italica L. with an identity of 99%. Theoretical model of the tertiary structure shows that it has a highly conserved containing the NPA motif which was further validated by Ramachandran plot. Analysis for cis regulatory elements identified more than 250 cis-regulatory elements present in the promoter region of PIP2;7 aquaporin gene in foxtail millet which revealed the presence of a number of abiotic stress responsive acting elements such as, ABRE binding site, MYB binding site, AP2/ERF binding site, LEA-5 binding site, NF-YB/A/C binding site, Trihelix binding site, EIN3:EIL binding site etc. Promoter analysis also revealed the presence of binding site for a sequence-specific DNA-binding domain (designated CG-1) present in calmodulin-binding transcription activators (CAMTAs) in the upstream region of PIP2;7 aquaporin. This domain could bind DNA directly and activate transcription, or interact with other transcription factors, not through DNA binding, thus acting as a co-activator of transcription. This might be the reason for high expression of PIP2;7 aquaporin. Applying different physiological and biochemical tests to appreciate drought tolerance in plant leads to faster selection methods. Therefore, these characters can be used as an indirect selection criterion for screening drought tolerance plant materials which will lead to new cultivars with high yield potential and high yield stability that in turn will result in superior performance in dry environments. Hence, this research can provide documentation for breeding/selection of higher drought resistant foxtail millet in arid regions and acquisition of good information for future molecular research. The physio-biochemically screened accessions can further prove to be useful in studies involving transcriptome changes to fetch out the molecular mechanism underneath of its drought adaptation. Accomplishments of these goals permit better understanding of molecular and physiological mechanisms utilized by plants during adverse growth conditions and define the biomarkers of tolerance against stresses. The research provides a possible point of integrating various molecular and biological pathways with water stress regulated gene expression. The study can prove helpful to the farmers in selecting foxtail millet cultivars for unaffected yields in diverse agronomic conditions. These findings provide insight for further investigation of CG-1 domain in plant aquaporins in opening new perspectives for improving drought tolerance which could eventually lead to better crop production. The identified aquaporin could be explored in development of tolerant lines by MAS using PIP 2;7 as a functional marker and also in transgenic development in other related crops.ThesisItem Open Access DNA BARCODING AND FINGERPRINTING OF CITRUS SPP.(Dr. Panjabrao Deshmukh Krishi Vidyapeeth, Akola, Maharashtra., 2022-09-23) CHUNGADA, ANIL SANDUSING.; Gahukar, Dr. S. J.The present investigation entitled “DNA Barcoding and Fingerprinting of citrus spp.” was carried out at Biotechnology Centre, Department of Agricultural Botany, Post Graduate Institute, Dr. Panjabrao Deshmukh Krishi Vidhyapeeth, Akola. Commercial citrus cultivation has been done by grafting and budding. The rootstock is a very important part of a citrus orchard. The contamination in source seed for the raising of the nursery of citrus rootstock is a major challenge in assuming the quality of orchard and ultimately yield of citrus. Thus a reliable and life stage-independent method of species identification is important. In the present investigation, 11 different SCAR markers were validated for discriminating Galgal rootstock from other citrus species under study. The DNA amplification was carried out with 11 SCAR markers. The primer-1, primer-2, primer-3, primer-4, primer-5, primer-6 and primer-11 show the band size of 283 bp, 380 bp, 172 bp, 310 bp, 150 bp, 137 bp and 300 bp respectively. All these primers are promising primers for discrimination of Galgal from other species under study. A total of twenty-one SSR primers were used to evaluate the genetic diversity of six citrus species. Seven primer pairs failed to show any amplification thus revealing no bands (null allele) in all the genotypes. Fourteen SSR markers exhibited polymorphism and showed high levels of allelic diversity. A total of 139 amplicons were amplified by 14 polymorphic SSR loci and the number of amplicons ranged from 5 to 18 with an average of 9.92 amplicons per locus. Only one primer i.e. SSR-5 shown 100 % polymorphism, whereas, 87 % polymorphism was observed in SSR-2 and 67 % polymorphism was detected in SSR-3, SSR-9, SSR-12 and SSR-13; showed an average 66.14 % polymorphism percent. Total alleles per locus were 9.92, whereas, the average number of monomorphic and polymorphic alleles are 2.92 and 6.85, respectively. The PIC value of 14 microsatellite loci ranged from 0.41 to 0.86 with an average value of 0.59. For each marker, the maximum PIC value was observed in marker SSR-5 i.e. 0.86 and the minimum was in SSR-11 i.e. 0.41. The present investigation also aimed to generate sequences for DNA barcodes of various citrus species by using matK, rbcL, ITS and trnH–psbA. The six citrus species i.e. Galgal (Citrus pseudolimon), Rangpur lime (Citrus limonia), Alemow (Citrus macrophylla), Jambhiri (Citrus jambhiri), Orange (Citrus reticulata) and Sweet orange (Citrus sinensis) were selected from All India Co-ordinate Research Project on Citrus, Dr. P.D.K.V. Akola and Central Citrus Research Institute, Nagpur. The isolated DNA was amplified with three plastids (rbcL, matK and trnH-psbA) and two nuclear internal transcribed spacer (ITS) gene regions. The rbcL, matK, and trnH-psbA shows the band size of 700bp, 1700bp and 625 bp respectively and ITS shows a band size of 400bp. The amplified PCR products were sequenced by using the Sanger sequencing method by using Applied Biosystem Sequencer from Eurofins Genomics India Pvt. Ltd. Bangalore. The rbcL gives a good sequence of 629 bp for Sweet orange (Citrus sinensis). Barcode was generated from this sequence on BOLD (Barcode of Life Database) by using its “Phd.1”, “Trace”, “ABI” file of the sequence having sample ID- SOR-1 and Process ID-SOR001-19. The Barcode generated by using the sequence of Jambhiri (Citrus jambhiri) 860 bp, amplified by universal marker matK by using “Trace”, “ABI” file of the sequence on BOLD system 3 (Barcode of life Database) having Sample ID CJ-1 and Process ID-GRJ003-20. The Barcode generated by using the sequence of Rangpur lime (Citrus limonia) 738 bp, amplified by universal marker matK by using “Trace”, “ABI” file of the sequence on BOLD system 3 (Barcode of life Database) having Sample ID CRL-4 and Process ID-GRJ004-20. The Barcode generated by using the sequence of Galgal (Citrus pseudolimon) 820 bp, amplified by universal marker matK by using “Trace”, “ABI” file of the sequence on BOLD system 3 (Barcode of life Database) having Sample ID CG-2 and Process ID-GRJ002-20. From the present study involving four different barcode regions under study, it was concluded that rbcL and matK markers are good for the generation of a barcode in Citrus Sinensis, Citrus Jambhiri, Citrus Pseudolimon and Citrus Limonia. DNA barcoding can be a very effective tool to identify citrus species. Here, we tested DNA barcodes of plant plastid DNA, rbcL, matK, trn H-psb A and internal transcribed spacer (ITS) to resolve available citrus species. For the single barcode region, matK had the highest rate of correct identification than rbcL. In the present work, we lay the foundation towards DNA barcoding applications for citrus plants. matK is proposed to be a suitable candidate DNA barcode marker for citrus species identification. Further need to explore with additional markers which may improve citrus species identification for practical conservation. The most valuable finding of this study is that the developed barcodes will help in the identification of quality rootstock for citrus propagation, which can increase the life of citrus orchards and the productivity of citrus.ThesisItem Unknown EFFECT OF DESICCATION ON CALLUS CULTURES IN COTTON(Dr. Panjabrao Deshmukh Krishi Vidyapeeth, Akola, Maharashtra., 2009-06-12) ZAMBRE, SUPRIYA MANOHAR; VAIDYA, E.R.ThesisItem Open Access GENOME WIDE ANALYSIS OF DNA METHYLATION IN RESPONSE TO MACROPHOMINA PHASEOLINA (CHARCOAL ROT) IN SOYBEAN(Publisher : Dr. Panjabrao Deshmukh Krishi Vidyapeeth, Akola, Maharashtra., 2022-01-08) MADKE, MEGHA SHAMRAO; Jadhav, Dr. P. V.Abstract : The exploration entitled “Genome wide analysis of DNA methylation in Macrophomina phaseolina (charcoal rot) in soybean (Glycine max L.)” was carried out during 2019-20 in Biotechnology Centre, Department of Agricultural Botany, Dr. PDKV, Akola to look over the extent and pattern of genomic methylation in response to charcoal rot disease in soybean genotypes. Variation in DNA methylation enables plants to inherit traits independently of changes to DNA sequence with the help of restriction enzymes that leads to amplify small region of genome. Here, for validation purpose we screened six selected resistance and susceptible soybean genotypes in response to charcoal rot diseases. Charcoal rot disease caused by Macrophomina phaseolina is one of the most damaging diseases of soybean resulting to 70 % losses and till date no immune genotype is known for the same. Various limitations inherent in field experiments may have hindered progress toward identifying new source(s) of resistance and breeding resistance in soybean. The Methylation Sensitive Amplified Polymorphism which is modification of Amplified Fragment Length Polymorphism (AFLP) used to investigate the epigenetic changes associated with the Charcoal rot in Soybean. The samples of genomic DNA were double digested with either HpaII or MspI and with EcoRI. To analyze epigenetic changes, 21 MSAP marker with different primer combinations screened and 11 primer combinations showed polymorphism at methylated sites in response to genotypic variation. A total of 312 and 311 clear and reproducible amplicons were amplified from control and CR treated samples, respectively. Under the control conditions, the total methylation of CCGG sequences averaged 49.03% in control and 39.22% in CR treated. The extent of DNA methylation in resistant cultivars were higher under control condition and lower in diseased condition. It means Charcoal rot decreased the percentage of total methylated amplicons in resistant cultivars, but increased it in susceptible genotype TAMS-38 (avg of 41.17%), when levels of DNA methylation were compared with those in the respective control (22.22%). These findings indicated opposite effects of disease on DNA methylation in genotypes of Soybean with different levels of resistivity to Charcoal Rot The fully methylated loci were always more abundant than the hemi-methylated ones. PCoA (Principle Co-ordination Analysis) and NJ (Neighbour Joining) radial tree diagram showed the dissimilarity between primers according to the scored data by genotypes. These results showed that there are great difference in genomic DNA of different soybean genotypes related to the response to the charcoal rot disease. The study revealed that methylation status varied substantially during the stressed or diseased condition. Polymorphic bands can be eluted and sequence further.ThesisItem Open Access IN VITRO SHOOT TIP GRAFTING IN CITRUS FOR QUALITY SEEDLING PRODUCTION.(Dr. Panjabrao Deshmukh Krishi Vidyapeeth, Akola, Maharashtra., 2017-10-10) MALKOTE, MANIK MADHAVRAO.; Akhare, Dr. A. A.The present investigation entitled “In Vitro Shoot tip grafting in citrus for quality seedling production” was conducted at Centre of Excellence In Plant Biotechnology, Dr. Panjabrao Deshmukh Krishi Vidyapeeth, Akola, Dist.Akola (M.S.) during the academic year 2015-2017 with the objectives to study the effect of different parameters influencing shoot tip grafting efficiency and efficacy. The experimental material of present investigation comprised the three varieties of citrus viz., Rangpur lime, Jambhiri as rootstocks and Sweet orange as scion. For micropropagation of Sweet orange field collected nodal explants and epycotyle segments from in vitro grown seedlings was used. The multiples obtained were then used for grafting purpose on Jambhiri and Rangpur lime. When seeds of Jambhiri, Rangpur lime and Sweet orange were inoculated for germination on various media combinations, The highest % of germination was found on MS media suplimented with 6% sucrose along with GA3 @ 1mg/l. The highest percent germination in case of Jambhiri and Sweet orange was 86.66% and in case of Rangpur the precent germination was 80.33%. For the preparation of scion, micropropagation of Sweet orange explants viz. nodal explants from field and epicotyls segment from in vitro germinated seedlings were tried on various concentrations of BAP @ 1, 2, 3 mg/l and NAA @ 0.5 and 1 mg/l alone or in combinations. In case of field collected nodal explants the highest success percentage was 66.66% in 6 days where as vitro derived explants respond best on media combination MS + BAP @ 3mg/l in 6.33 days. In order to optimize shoot tip grafting protocol various parameters such as sucrose concentration, age of rootstocks effect of PGRs, size of scion etc. were standardized. Amongst which sucrose % from 1.5 to 6% were tried, out of which 6% sucrose showed highest number of successful grafts on both the rootstocks Rangpur lime and Jambhiri. The rootstock age was tried from 5 to 25 days after germination and highest % success 71.42% and 62.50% in Jambhiri and Rangpur respectively was recorded when 15 days old rootstocks was used. Different PGRs viz. kinetin (1 mg/l and 5 mg/l) and BAP (1mg/l and 5mg/l) were tried in order to enhance graft union and scion growth, the highest % success of graft was noted when scion were treated with BAP @ 1mg/l prior to graft it on rootstock either Rangpur lime (66.66%) Jambhiri (73.33%). The scion size tried varies from 0.2 to 0.7 cm amongst which the highest % of success was shown by 0.6 to 0.7 mm scions. In case of Rangpur lime the % success of graft was 58.33% where as 8.033% success was recorded in Jambhiri rootstocks.ThesisItem Open Access IN-PLANTA TRANSFORMATION OF SOYBEAN (Glycine max L.) TO ENHANCE SALINITY AND DROUGHT TOLERANCE(Dr. Panjabrao Deshmukh Krishi Vidyapeeth, Akola, Maharashtra., 2017-10-10) LUNGE, AKSHAY RAJENDRA; Jadhav, Dr. P. V.In the present investigation entitled “In-Planta transformation of soybean (Glycine max L.) to enhance salinity and drought tolerance” was under taken with the view to develop In-planta regeneration system for elite soybean genotype. The study was carried out at Biotechnology Centre, Dr. PDKV, Akola during year 2015-16. In vitro culture and genetic transformation of soybean is difficult due to its recalcitrant nature. Establishment of gene transfer procedure is prerequisite to devolved transgenic plant of soybean in a shorter period. Therefore, the transformation was perform using the most popular soybean cv., JS-335 to optimize the factor influencing transformation efficiency through Agrobacterium tumefaciens based in-planta transformation using LBA4404 strain harboring gene, PDH45. In the present study four strategies were employed for in-planta transformation i.e. infection to seed, infection to seed and then incubation for 24 h in Agrobacterium, infection to every developing stage and infection to flowering and pod filling stage. Several parameters, such as optical density, incubation time, acetosyringone concentration influencing In-planta transformation, have been evaluated in this study. In seed radicle treatment, number of flower and pod formation was highest among the treatments. All the plant samples (T1) tested were found to be negative after PCR confirmation with gene specific primer. Similarly, in treatment injected to seed radicle and incubated for 30 min. out of 50 sample two samples were found to be positive after PCR analysis with gene specific primer. Putative transgenics were screened by PCR confirmation. It helped to eliminate non-transgenic. The method also decreased the time and effort involved in the screening process The procured plasmid containing PDH 45 gene was transformed into the Agrobacterium using In-planta method without using helper strain and confirmed by restriction digestion and the gene specific set of primers.ThesisItem Open Access INTROGRESSION AND CHARACTERIZATION OF BACKCROSS POPULATION OF SORGHUM.(Dr. Panjabrao Deshmukh Krishi Vidyapeeth, Akola, Maharashtra., 2018-02-06) RAUT, DIMPAL SHESHRAO.; Gahukar, Dr. S. J.The present investigation entitled “Introgression and characterization of backcross population of sorghum” was carried out at Molecular Breeding Laboratory, Biotechnology Centre, Department of Agricultural Botany, Post Graduate Institute, Dr. Panjabrao Deshmukh Krishi Vidyapeeth, Akola in academic year 2015-2016. The study was aimed at introgression the glossiness and trichome density resistant QTLs using specific SSR markers. A total 80 BC4 plant progenies were obtained by crossing AKSV-13R X IS18551, all these progenies were evaluated for phenotyping and genotyping using specific foreground and background primers. Wide morphological variation was observed among the genotypes for the shoot fly resistance component traits viz., pigmentation, leaf glossiness, seedling with eggs, dead hearts, trichome density and yield. Phenotypic correlation between the component traits of shoot fly resistance were estimated based on individual progeny from each of BC4 population under study. Yield showed highly significant positive association with trichome density in BC4 population under study. These traits could be utilized as simple criteria for phenotypic selection for resistance to shoot fly in sorghum. Foreground selection of BC4 progenies using markers associated with genomic region for glossiness and trichrome density revealed specific donor parent type alleles in plant no. BC4-2, BC4-10, BC4-14, BC4-17, BC4-19, BC4-25, BC4-54, BC4-60, BC4-76, BC4-79 from cross AKSV-13 X IS18551. Background selection of BC4 revealed that recurrent parent type of in plant no. BC4-1, BC4-6, BC4-13, BC4-17, BC4-35, BC4-41, BC4-43, BC4-55, BC4-68, BC4-73 population of genome recovery of recurrent parent. Amplification with foreground markers revealed specific allele recovery from donor parent (IS18551) in BC4 population. Population plant no. BC4-3, BC4-50 in foreground markers and plant no. BC4-5, BC4-70 in foreground markers showed maximum alleles with expected size.ThesisItem Open Access INTROGRESSION STUDIES OF FUSARIUM WILT RESISTANCE IN PKV KABULI 4 USING MOLECULAR MARKERS.(Dr. Panjabrao Deshmukh Krishi Vidyapeeth, Akola, Maharashtra., 2021-10-31) KHELURKAR, VAIBHAV CHANDRAKANT.; SAKHARE, Dr. S. B.The present investigation entitled “Introgression studies of Fusarium wilt resistance in PKV Kabuli 4 using molecular markers” was carried out at the Biotechnology Center and Pulses Research Unit, Department of Agricultural Botany, Dr. PDKV, Akola. The MABC program was initiated with the cross between PKV 4 (Moderately susceptible, recurrent parent) and WR-315 (Resistant, donor parent) to develop Fusarium wilt resistance to foc-1 during crop season September 20018-19. Total 67 different molecular markers were used for the parental polymorphism survey and seven markers found polymorphic namely, TA 194, TS 82, TA 59, TA 110, showed 100 % polymorphism and TR 19, STMS 02, TA 03 showed 75 % polymorphism. The identified polymorphic markers help in differentiating male and female parental lines and successfully detect the F1 hybrid progenies. From the hybridization program 75 F0 were seeds obtain Among 75 F1s plants, 45 hybrids were confirmed through using polymorphic markers (TA 14, TA 194, and TR 19). Further, the confirmed hybrids were used for the execution of the first round of backcrossing and 18 BC1F1 plants were grown at this point foreground and recurrent parent genome recovery was carried out. Foreground selections were carried out using linked markers namely, TA 59, TA 110, TA 194, TR 19 for selection of introgressed target region and six common heterozygous plants were selected. In BC1F1 population selection for target, the region was carried out using the linked markers. For identification of Fusarium wilt QTLs in the F2 population, 60 F2 individuals from a single cross were used simultaneously for genotyping by six polymorphic markers and phenotyping in the artificial greenhouse sick pot conditions. Using morphological and molecular data linkage map was constructed and one major QTL “qfwch-02” were identified. Which covered a total map length of 93.60 cM, the average map distance between any pair of markers was 16.1 cM and major QTL “qfwCh-02” was identified on linkage group 2 for Fusarium wilt resistance against race-1 at map position 31.4 cM in F2 progenies of PKV Kabuli 4 X WR-315. This QTL explained 10.91 % phenotypic variance with LOD score 3.3 flanked by left marker TA 110 and right marker TR 19. Background selection was performed in 18 BC1F1 individuals using 20 molecular markers distributed throughout the chickpea genome. The range of RPGR analysis in BC1F1 was ranged from 57 % (BC1F1-180) to 67.5 % (BC1F1-17). During the present investigation the validated markers, for foreground selection namely, TA 100, TR 19, TA 194, TA 59 for foc-1 race will help in early detection of resistant and susceptible plants in chickpea mapping population and to introgress fusarium wilt resistance in chickpea genotypes by Marker Assisted backcross breeding program. Developed genomic linkage map and QTLs identified will be useful for chickpea genetics and breeding applications. Moreover, linked markers found with identified QTLs for FW race-1 will be useful for molecular breeding for Fusarium wilt chickpea improvement.ThesisItem Open Access MOLECULAR AND BIOCHEMICAL CHARACTERIZATION FOR WATER STRESS TOLERANCE IN WHEAT GENOTYPES(Dr.Panjabrao Deshmukh Krishi Vidyapeeth, Akola., 2017-01-18) TANDALE, RAM MHATARDEO; Gahukar, Dr. S. J.Drought affects crop growth and yield by altering metabolic and physiological processes. Plants tolerate drought through osmotic adjustment, stomatal resistance, increased water uptake and accumulation of waxy layer. Cultivars possessing drought tolerant traits may be promising candidates for drought prone regions. The present study was planned for molecular and biochemical characterization for water stress tolerance in wheat genotypes. In this study various morphological, physiological, biochemical and molecular observations were recorded. Drought stress was created with different concentrations of PEG-6000 (-2, -4, -6, and -8 bars) and by withholding of water at moderate and sever level. On the morphological basis, all cultivars exhibited decline in germination percentage, speed of germination, shoot-root length, coleoptile length, seedling fresh and dry weight under water stress environments. While on biochemical basis proline has direct relationship with osmotic stress and water stress while total chlorophyll content has inverse association with osmotic stress and water stress. On the physiological basis, with increase in water stress photosynthesis rate, relative water content decrease while epicuticular wax content increased with an increase in osmotic stress and water stress conditions. All these parameters finally extremely affect the yield of wheat genotype. At the end for drought response, wheat cultivars were evaluated on molecular basis. A set of trait specific 18 SSR primers were used for identification of drought tolerant genotypes. From which 16 primers gave polymorphic result and amplified unique band as result of drought tolerance. AKW-381 followed by AKAW-4627, AKAW-4210-6, AKAW-4498 and AKAW-4842 performed best at various stress levels for germination percentage, speed of germination, root-shoot fresh and dry weight, total chlorophyll content, proline, Rate of photosynthesis, wax content, and relative water content. The performance of HD-2189, GW-322, NIAW-917 and AKAW-3722 was poor. On molecular basis expected size unique band was amplified in the tolerant genotypes AKW-381, AKAW-4627, AKAW-4210-6, AKAW-4498 and AKAW-4842. It can be concluded that water stress levels had substantial effect on germination and seedling growth. Morphological, biochemical, physiological and molecular analysis revealed that adequate genetic difference for drought tolerance existed in the tested genotypes. AKW-381 and AKAW-4627 may prove a promising parent material for breeding drought tolerant wheat cultivars. Further molecular investigations are suggested to assess the genetic basis of drought tolerance. AKAW-4627, AKAW-4210-6, AKAW-4498 and AKW-381 may be considered better genotypes for low rainfall or drought prone areas.ThesisItem Open Access MOLECULAR AND BIOCHEMICAL RESPONSES OF WHEAT GENOTYPES (Triticum aestivum) UNDER WATER STRESS.(Dr. Panjabrao Deshmukh Krishi Vidyapeeth Akola, Maharashtra., 2021-08-27) SURBHAIYYA, SHOBHA DEVIDAS.; Gahukar, Dr. S.J.The present investigation entitled “Molecular and biochemical responses of wheat genotypes (Triticum aestivum) under water stress” was carried out at the Biotechnology Centre, Department of Agriculture Botany, Dr. Panjabrao Deshmukh Krishi Vidyapeeth, Akola during 2016-2019. In the present study, the objectives were accomplished at two phases of environment, physiological screening under in-vitro (lab) condition and morpho-physiological, biochemical, molecular screening under in-vivo (green house and field) condition. The experiments comprised of eight genotypes with two check viz., AKAW-3717 (tolerant) and AKAW-3722 (susceptible). In first phase of experiment, osmotic stress imposed under laboratory condition by using different concentration of PEG- 6000 viz., 0, 15 %, and 25 % and performances of various genotypes were monitored against a control. Seedling traits such as germination percentage, seedling vigor index, shoot length, root length, shoot fresh/dry weight and root fresh/dry weight were studied in unstressed and stressed condition. The result indicated that increase in osmotic stress caused a significant decreased in above seedlings parameters. Correlation coefficient studies revealed considerable and positive correlation among seedling traits.The result indicated that decrease in one trait may cause simultaneous decrease in other traits; hence, selection for any of these seedling attributes will lead to develop water stress tolerant wheat genotypes. Based on these observations, most water stress tolerant and susceptible genotypes were selected. (Check tolerant) AKAW-3717, AKAW-4842, and AKAW-5017 were recorded as the most water stress tolerant genotypes, whereas, (Check susceptible) AKAW-3722, AKAW-4925, and AKDW-5012 were recorded as susceptible genotypes. In second phase of experiments (under green house and field condition), the extent of yield reduction with water deficit depends not only on the magnitude of water deficit but also on the stage of plant growth at which it develops. Under green house condition, a pot study water stress treatments were created by using different concentration of PEG-6000 (viz., -3 bar and -6 bar PEG-6000) and water withholding. Under field condition, water stress were created by only withholding of water to assess the effect of induced water stress on performance of ten wheat genotypes at two critical growth stages (tillering and flowering). After seven to fourteen days, plants exhibited visible effects of stress. Thus plant sample were collected for further analysis and data collection. Screening of wheat genotypes for water stress tolerance on the basis of morphological, physiological, biochemical, and molecular level. The results showed that water stress significantly reduced in morphological and physiological traits like plant height, total tillers, spike length, number of spikes per plant, number of grains per spikes, 1000 grain weight, yield per plant, relative water content, and chlorophyll content. The biochemical analysis revealed increased total soluble sugar, total soluble protein, and proline significantly with increasing water stress. Proline content in stressed tolerant plants is found to be very higher as compared to that in unstressed susceptible plants suggesting its key role in water stress tolerance in plant. The activity of CAT was found to be highest in AKAW-3717 at both tillering and flowering growth stages which was found to a tolerant genotype in prior morphological and biochemical screening. The activity of POD was found maximum in all tolerant genotypes viz., AKAW-3717, AKAW-4842, and AKAW-5017 than in susceptible genotypes viz., AKAW-3722, AKAW-5010, and AKAW-4926. Superoxide dismutase activity was found to be highest in AKAW-5017. Water stress preferentially enhanced the activities of enzymatic antioxidant and osmolytes. Differential transcriptome analysis using cDNA based start codon targeted polymorphism (SCoT marker) and oligodecamer (RAPD marker) were accomplished To identify differentially expressed TDFs. TDFs (cDNA-SCoT profiling) were produced in unstressed and stressed plant at tillering and flowering stages. At tillering stage, 186/215 (green house/field) TDFs were found differentially regulated out of 191/218 TDFs while 171/228 TDFs were found differentially regulated out of 177/229 TDFs at flowering stage. cDNA-SCoT profiling revealed that marker SCoT 03, SCoT 05, SCoT 11, SCoT 13, SCoT 14, and SCoT 18/SCoT 01, SCoT 04, SCoT 11, SCoT 14, SCoT 18, and SCoT 20 showed 100 % polymorphism at both stages. Similarly, TDFs (cDNA-RAPD profiling) were produced in unstressed and stressed plant at tillering and flowering stages. At tillering stage, 381 TDFs were found differentially regulated out of 398 TDFs while 373 TDFs were found differentially regulated out of 384 TDFs at flowering stage. AT both stages, cDNA-RAPD profiling revealed that marker OPF 7, OPF 14, OPH 16, OPB 10, OPI 16, OPI 13, OPI 2, and OPH 12 showed 100 % polymorphism. Further, gene expression studies were carried out using the four contrasting genotypes AKAW-3717 (check tolerant), AKAW-3722 (check susceptible), AKAW-4842, AKAW-4925, and AKAW-5017. Under green house as well as field condition there are five water stress specific markers (WDHN 13, α-Tubulin, WPIP, WTIP 11, and DREB 1A) were used for presence and expression of gene which can confer water stress resistance to a genotype. RNA was extracted by TriZol method. Then first strand cDNA synthesis was done by using Himedia cDNA synthesis kit. PCR was carried out by using the cDNA. PCR products were separated on polyacrylamide gel and visualized under gel doc system. AKAW-3717 followed by AKAW-5017 and AKAW-4842 performed best at various stress levels for morpho-physiological and biochemical parameters. However performance of AKAW-3722, AKAW-5010 and AKAW-4926 was poor. The gene expression, results indicated that, α-Tubulin showed their expression in all genotypes grown under controlled as well as water stressed conditions. While, WDHN 13 showed their expression in AKAW-3717, AKAW-5017, and AKAW-4842. None of the other gene expression was recorded in any other genotypes. It can be concluded that water stress levels had substantial effects on germination and seedling growth. Morphological, biochemical, physiological and molecular analysis revealed that adequate genetic difference for water stress tolerance existed in wheat genotypes tested AKAW-3717 and AKAW-5017 may prove a promising parent material for breeding water stress tolerant wheat genotype. Further molecular investigations are suggested to assess the genetic basis of water stress tolerance. AKAW-3717 and AKAW-5017 may be considered better genotypes for low rainfall drought prone areas. The present study can provide clues in identifying candidate genes for further functional analysis to delineate their precise role in abiotic stress response. As key genes are identified, efficiency increase and opportunities for genetic engineering are realized. This is a fundamental aspect of research into abiotic stress tolerance, and discoveries of abiotic stress tolerance genes, which is explored in the present study.ThesisItem Open Access MOLECULAR AND GENETIC ANALYSIS OF PUTATIVE TRANSFORMANTS IN TRANSGENIC PIGEONPEA.(Dr. Panjabrao Deshmukh Krishi Vidyapeeth, Akola, Maharashtra., 2017-12-05) KADAM, KARTIKI DADASAHEB; Akhare, Dr. A. A.The gram pod borer (Helicoverpa armigera Hubner) is the most serious insect pest of pigeonpea. It is highly susceptible to the insecticidal proteins of Bacillus thuringiensis (Bt). A codon-optimized chimeric Cry1Aabcgene of Bt driven by a constitutive promoter was introduced pigeonpea (PKV Tara). Molecular analyses were carried out by most of worker in T1 and T2 generation. From 7 plants of T3 generation total 57 progenies were tested for PCR from which 47 plants were seen to be positive through PCR analysis. 38 plants of T1 generation from DK+ material were tested for PCR analysis through gene specific primers from which 22 plants were seen to be positive. Kamble in year 2014-15 attempted 130 embryo infections with Agrobacterium strain EHA105 harboring Cry1Aabc and NptII gene out of which 73 were survived but only two plants were confirmed positive through PCR. T1 and T2 generation of these two plants namely A and B were sown. Three plants from eachwere selected for PCR confirmation. From two positive plants total 6 progenies from T1 generation and 3 plants from T2 generation were tested for PCR analysis all the 6 plants from T1 generation were seen to be positive while from T2 generation out 3 only two plants were positive. Sample population size of T1 and T2 plants was too less to study inheritance pattern. In order to study Mendalian inheritance pattern in T3 progeny PCR analysis for the amplification of the cry1Aabc gene and NptII gene was carried out. The plants from same parents plant was consider together for the inheritance study. For undertaking the Chi-square test for segregation the minimum population size of 6 was taken. Calculated x2 value is less than table value in all the plant progenies. Thus plants for the amplification of the Cry1Aabc gene and NptII gene and progeny of each clone shows 3:1 (presence: absence) segregation pattern. For detached leaf bioassay 19 positive plants and one negative plant which were confirmed by DNA extraction and PCR amplification with NptII and Cry1Aabc gene specific primers were tested using leaf feeding assay with neonate larvae of Helicoverpa armigera. 20 putative transformants of pigeonpea were subjected to insect (Helicoverpa armigera) bioassay using neonates of Helicoverpa armigera and young trifoliate leaves and WT (non-transformed) PKV Tara was used as control. Thus the result revealed that the level of Cry protein expressed in positive plants were able to reduce the leaf damage. Thus, it was concluded from above results that like positive plants selected for bioassay study having potential for providing tolerance against Helicoverpa armigera. The further confirmation in subsequent generation is required to come to more robust conclusion.ThesisItem Open Access MOLECULAR AND SEROLOGICAL DIAGNOSTIC METHOD FOR THE ASSESSMENT OF TRISTEZA VIRUS IN CITRUS SPP.(Dr. Punjabrao Deshmukh Krishi Vidyapeeth, Akola, Maharashtra., 2020-03-18) THAKRE, RITESH ASHOKRAO.; Gahukar, Dr. S. J.The present investigation entitled “Molecular and serological diagnostic method for the assessment of tristeza virus in citrus spp.” was carried out at Biotechnology Centre, Department of Agricultural Botany, Post Graduate Institute, Dr. Panjabrao Deshmukh Krishi Vidhyapeeth, Akola. Traditional method of Citrus tristeza virus (CTV) detection like surveys and biological indexing are relatively less reliable, time consuming and not feasible economically, preclude the handling of large number of samples beside of this molecular and serological methods has revolutionized the detection test, making it feasible to test large number of samples, which provide rapid, sensitive and economical detection of CTV in crude extracts. Hence the present investigation was designed to developed and validate the molecular and serological diagnostic methods for CTV detection at very early stage of infection to prevent the pathogenic spread. For molecular diagnosis of CTV, viral RNA was isolated from the CTV infected leaf samples of citrus spp. showing characteristic symptoms of CTV as vein clearing, yellowing, leaf cupping, quick decline etc. were collected from two location of Vidarbha region, during July-December in 2016-17 of Maharashtra state as well as from two different growth stage of citrus spp. using trizol reagent method. Polymerase chain reaction was carried out with ease of coat protein gene specific primers, showed amplification at 672 bp, confirmed prevalence of CTV. BTC series primers were validated to check cross amplification with other citrus infecting viruses. No amplification observed in other viruses samples and also in non template control, revealed that BTC series primers amplifies only CTV isolates from citrus field and hence it can be used in diagnosis and certification programme. In serodiagnostic approach CTV was isolated and purified from infected citrus spp. samples using ultracentrifugation method by caesium sulphate and sucrose gradient. Electron microscopic studies of the purified CTV showed the flexuous rod shaped particales measuring 2000 nm in length and 12 nm in diameter. In biological indexing of CTV, indexed seedlings produced characteristic disease symptoms of leaf cupping, vein clearing and yellowing, after two to four month of inoculation and subsequent growth was reduced, leaving the plants with a dwarfened appearance, shorter stem, internodes, petioles, and underdeveloped. Polyclonal antibodies specific to CTV were produced in Rabbit using ultrapurified virus and raised antiserum specificity was confirmed with Dot-Blot. Polyclonal antibodies detected closely related species during antibodies specificity confirmation. In Dot-Blot of Orange CTV sample, titter dilution was recorded at 1:10 of antigen 1:2000 of primary antibody and 1:1000 of secondary antibody. In sweet orange it was recorded at 1:20 of antigen 1:2000 of primary antibody and 1:1000 of secondary antibody, in lemon also, it was 1:20 of antigen 1:2000 of primary antibody and 1:1000 of secondary antibody. No colour was developed in control indicated sensitivity of assay. In Orange, Enzyme Link Immunosorbent Assay (ELISA) OD reading was recorded at 405 nm for the detection of CTV. Optimum OD reading 1.31 was observed in the combination of dilution of 1:20 antigen and 1:4000 dilutions of antisera with secondary antibody dilution of 1:1000. Optimum OD 1.26 was observed in the combination of 1:20 dilution of antigen and 1:4000 dilution of antisera with secondary antibody dilution 1:2000. Similarly, in sweet orange optimum OD reading 1.35 was observed in the combination of dilution of 1:10 antigen and 1:16000 dilutions of antisera with secondary antibody dilution of 1:1000. Optimum OD reading 1.32 was observed in the combination of 1:10 dilution of antigen and 1:8000 dilutions of antisera and with secondary antibody dilution of 1:2000. Also, in case of lemon optimum OD reading 0.54 was observed in the combination of dilution of 1:20 antigen and 1:4000 dilutions of antisera with secondary antibody dilution of 1:1000. Optimum OD reading of 0.51 was observed in the combination of 1:20 dilution of antigen and 1:4000 dilutions of antisera and with secondary antibody dilution of 1:2000. Among the three citrus spp. sweet orange was heavily infected with CTV according to conclusion drawn from both DOT-BLOT and ELISA serological method. Plate ELISA at higher antibody and antigen dilutions was performed to check the reactivity at lowest concentration of antibody and antigen. No colour development was observed in any combinations and all OD readings were negative in the ELISA reader at 405 nm, indicate that dilutions of antibody beyond 1:32000 and dilutions of antigen above 1:50 were not working for detection of CTV Immuno diagnostic kit was developed and validated successfully by using different CTV infected citrus spp. samples. Out of eight CTV symptomatic samples selected morphologically, four showed incidence of CTV occurrence. Result of immunodiagnostic kit developed using polyclonal antibody proved to be efficient and reliable for CTV detection. Standardized and validated molecular and serological method of CTV detection can be used further for the CTV diagnosis in citrus spp. and also for certification programme.ThesisItem Open Access MOLECULAR ASSESSMENT OF SINGLE CROSS HYBRIDS OF DESI COTTON FOR YIELD AND BOLL RELATED TRAITS USING SSR MARKERS.(Dr. Panjabrao Deshmukh Krishi Vidyapeeth, Akola, Maharashtra., 2022-01-27) BHOJANE, PALLAVI NARENDRARAO.; RATHOD, Dr. D. R.Present experiment entitled “Molecular assessment of single cross hybrids of desi cotton for yield and boll related traits using SSR markers” was undertaken at Biotechnology Centre, Dept of Agril. Botany, Dr. PDKV, Akola during 2019-20 with objective to transfer important traits related to boll size and seed cotton yield to enrich gene pool of G. arboreum (desi) cotton using popular and released varieties viz., AKA-7 (P1), Phule Dhanwantari (P2) as a female parent, while donor parent’s G. cernuum (D1) and PA-812 (D2). Four single cross hybrids viz., SC-I (AKA-7xG.cernuum), SC-II (AKA-7xPA-812), SC-III (PD x G.cernuum) and SC-IV (PDxPA-812) attempted during 2018-19 were used further to develop possible two double cross (DC) and one multi cross (MC) hybrids. Accordingly, a total of 333 crosses were attempted consisting of 59, 102, 67 and 105 crosses of SC-I, II, III and IV respectively. Out of these, a total of 55 crosses made successful with 10, 18, 7 and 20, from crosses of SC-I, II, III and IV respectively. A total of 971 F1s seeds harvested individually with 190, 377, 131 and 273 seeds from single crosses SC-I, II, III and IV, respectively. From these single crosses, a total of 118 double cross seeds consisting of DC-I 52 seeds and DC-II 66 seeds were obtained. Harvested seeds from each individual crossed boll of four SC and two DC hybrids were raised for its phenotypic and genotypic evaluation during off-season nursery under polyhouse facilities. Simultaneously, raised seedlings were subjected for molecular assay to ensure its successful crossing. Mean performance of varietal features of parents were assessed under field condition for nine quantitative characters viz., days to 1st flower, days to 1st boll bursting, plant height, no. of monopodia, sympodia and bolls plant-1, boll weight and size, seed cotton yield plant-1. Morphological data recorded on five randomly selected plants confirms its varietal identity. Few SC and DC progeny lines found promising for boll wt. 3.70 -5.10g big boll size. During molecular characterization, parental polymorphism survey was undertaken using 70 microsatellite markers. Out of these 31 SSRs were not amplified and uninformative, whereas remaining 39 shows satisfactory level of amplifications. Among these 39 SSR amplified in parental population, only 20 SSR shows polymorphism among parents and their resultant SC hybrids. Remaining 11 SSR shows monomorphic bands, hence discarded. Among polymorphic 20 SSRs, only 8 SSR were able to distinguish SC and DC hybrids and only these markers used to characterize SC and DC hybrid progeny population. Remaining 12 SSR among 20 polymorphic observed, where found non consistent in hybrids, when tested in either SC or DC. The average number of alleles was found 3.44 primer-1 with highest polymorphism (94%). Among SSRs, only BNL-1694 and BNL-3580 showed highest level of polymorphism 94%. A total eight DC hybrids, out of 118 obtained in DC-I and DC-II were confirmed as true hybrids by SSR genotyping. These true hybrids were further evaluated for trait evaluation and introgression of gene in segregating progenies of each single cross and double cross hybrids.ThesisItem Open Access MOLECULAR CHARACTERIZATION OF USEFUL MUTANTS OF FOXTAIL MILLET (Var.PS4).(Dr. Panjabrao Deshmukh Krishi Vidyapeeth, Akola, Maharashtra., 2017-12-05) PADMANE, SATISH BHIKAJI; Moharil, Dr. M. P.The present investigation entitled “Molecular characterization of useful mutants of Foxtail millet (Var.PS4)” was under taken with the view to identify the promising mutants with respect to yield contributing traits. The study was carried out at Biotechnology centre, Dr. PDKV., Akola during year 2016-17. In the present study 50% lethality reduction was observed at 400 Gy treatment and hence it was (400 Gy) imposed for further irradiation with subsequent treatment of EMS with different time intervals ranging from 6 hrs to 24 hrs. The genotype of Foxtail millet Var. PS4 was irradiated with 400 Gy and 0.1% EMS. Treated seeds were sown at experimental field of Department of Botany to rise M1 and followed by M2 generation. The phenotypic observations with respective to yield contributing traits such as plant height, panicle number and density, anthocyanin coloration of bristles, reproductive tiller, distance of third internode, lethality, mutation frequency and mutagenic effectiveness were carried out M2 generation. However, in M1 generation observation on plant injury were recorded. Based on these phenotypic observations 110 promising mutants of foxtail millet for different attributes like low plant height, high bristles, high number of panicle, panicle length, reproductive tillers and plant yield were identified. The mutagenic concentration (400Gy+ 0.1%EMS, 24 hr treatment) resulted in significant increase in tiller numbers and panicle numbers, While resulted to be best treatment for dwarf height (400Gy) and (400Gy+ 0.1%EMS, 6hr treatment). Treatment for panicle length with significant increase is (400Gy+ 0.1%EMS, 18 hr treatment) and (600Gy). In this study, 27 SSR markers and 5 gene specific markers were evaluated in 9 foxtail millet genotypes. Promising mutants from M3 generation i.e. dwarf height, more number of panicles, number of reproductive tillers, panicle length, earliness and high bristles were selected for molecular screening. Molecular characterization using SSR markers and gene specific markers revealed considerable variability induced at various loci, targeted by SSR and gene specific markers. All 27 SSRs were polymorphic across the 9 foxtail millet genotypes. A total of 54 monomorphic and polymorphic alleles were detected with an average number of alleles of 1.87 and 0.62 per locus (range 1-2 per locus). Amongst 27 SSR markers used, SiGMS4069, SiGMS7828 and SiGMS2453 showed 100% percent polymorphism, followed by SiGMS12150 shows 66% polymorphism. SiGMS10547, SiGMS11633, SiGMS6281, SiGMS6938, SiGMS14131, SiGMS1995, SiGMS203, SiGMS9021 SiGMS9551 and SiGMS7002 shows 50% percent polymorphism respectively. Null promorphic observance were observed in gene specific markrers. The genetic variability analysis using jaccard’s coefficient clearly separated 9 foxtail millet genotypes into five distinct clusters in SSR markers and three distant clusters in gene specific markers were done. The genotypes belonging to the same cluster are genetically more or less similar and the genotypes belonging to the different clusters are genetically different from each other. The polymorphic marker identified in the present investigation for the characterization of promising mutants will be explored, in future, to see the association with any desired phenotype. Selective genotyping may be useful to see the association between genetic diversity and phylogenetic data, otherwise segregating population will have to screen. However, point mutations cannot be or very rarely detected by the SSR marker, considering this different approaches like single stranded confirmation polymorphism (SSCP), Endonucleolytic Mutation Analysis by Internal Labelling (EMAIL), High resolution melting (HRM), Heteroduplex analysis, should be used in future to investigate the important point mutation in functional gene. In the present investigation entitled.ThesisItem Open Access MORPHOLOGICAL CHARACTERIZATION OF JAVA CITRONELLA VARIANTS AND ENCAPSULATION OF OIL FOR SLOW RELEASE.(Dr. Panjabrao Deshmukh Krishi Vidyapeeth, Akola., 2017-10-10) GANGURDE, ANUJA SUKLAL.; Akhare, Prof. A. A.The present investigation entitled “Morphological characterization of java citronella variants and encapsulation of oil for slow release” was carried out at Biotechnology Centre, Department of Agricultural Botany, Post Graduate Institute, Dr. Panjabrao Deshmukh Krishi Vidyapeeth, Akola in academic year 2016-2017. Morphological characterization of five selected variants of in vitro mutagenized plants along with 4 promising control genotypes of java citronella i.e Bio-13, jalpallavi, medini and mandakini was done. On the basis of various qualitative character such as plant height(cm), number of tillers, leaf width (cm), leaf blade length (cm), yield parameter such as oil yield (100g/ml), biomass (g), morphological characters such as steam colour characterization was done. All the five variants of in vitro mutagenized plants have performed well for characters like plant height, number of tillers, leaf blade length and biomass but the variant 4 and 5 performed best over control genotypes for the character such as for plant height variant 4 (118.40) cm and variant 5 (113.60) cm, number of tillers (59.40),(61.93), leaf blade length (88.86) cm, (90.73) cm, Biomass (4204.47)g, (4526.69)g. While for characters like leaf width the variants were small as compared to control Bio-13 (2.0) cm, jalpallavi (1.78)cm High oil yield was recorded in variant 4, 5 (1.38), (1.40), Bio-13 (1.45), jalpallavi (1.42) 100g/ml. steam colour had shown variation from as green, light green, green brown, dark brown in variants and control genotypes respectively. Qualitative determination of oil constituents by thin layer chromatography was done by using different solvent system but among that Hexane : Ethyl acetate (80:20 v/v) has given best separation and identification of components .The components identified using this solvent system were geraniol with Rf value (0.77), (-)- citronellal (0.98), β-citronellol (0.80), (-)-β citronellol (0.81) with Greenish blue, Dark violet, Faint blue colour were obtained. The Oil in water (o/w) type chitosan encapsulated volatile Citronella Oil microcapsules were prepared with various concentration of chitosan ranging from (0.5 percent to 1.5 percent) and NaOH concentration ranging from (0.5 percent to 1.5 percent). The significant formlulation of microcapsules were obtained at 1.5 percent chitosan and 1 percent NaOH.The encapsulation effeciency of 95.61 percent was observed with storage stability upto 25 days.
- «
- 1 (current)
- 2
- 3
- »