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ThesisItem Open Access Molecular profiling of mungbean genotypes vigna radiata (L.) wilczek using RAPD analysis(CCSHAU, 2007) Madhu; Subhadra SinghA study on 39 genotypes of mungbean representing a cross-section of variability was undertaken to assess RAPD polymorphism and to estimate the matrix of Jaccard’s similarity coefficient. A total of 411 amplified products comprising 382 (92.9%) polymorphic and 29 (7.1%) monomorphics markers were observed using 30 RAPD primers. The number of bands generated per primer ranged from 7 to 20 bands with a mean of 13.7 bands per primer. The primers showed a large variation in the frequency of polymorphic bands (0.026 - 0.974) and in number of banding patterns (3 – 35). Efficiency of primers were analyzed using Discrimination power (D), Diversity index (DI), Resolving power (Rp) and Marker index (MI). All efficiency parameters except DI could identify most efficient (S-1 and S-2) and the least efficient (S-30) primers. Both discrimination power (D) and resolving power (Rp) parameters appear to be good measure of the usefulness of primers to identify cultivars. The experimental results obtained for real efficiency of the primer agreed well with that of theoretical expectation based on independent hydpothesis of primers. Genetic similarity coefficient ranged from 40.8 per cent to 90.3 per cent indicating moderate level of genetic variability among genotypes. Some of the genotypes such as K 851, Pusa Ratna, MH 98-1, Asha, MH 96-1 Muskan, Koppergaon, K 851, Pusa Baishakhi, ML 1108, PDM 262, ML 682, Pusa 9072 and ML 818 exhibited higher genetic similarity with rest of the genotypes. ML 131 was the most diverse genotype. Other genotypes namely ML1349, Pant M-4, Greengram Co-5, SML 32, UP 99-3, ML 1333, UPM-98 also exhibited higher diversity. The dendrogram constructed from Jaccard’s similarity coefficient showed three main groups. The group A consists of 26 genotypes while group B and C consist of three and two genotypes only. The mungbean genotypes evolved at PAU, Ludhiana (Punjab) and in Uttar Pradesh were comparatively more diverse among themselves than the group of genotypes from Hisar (Haryana) and IARI (Delhi). The grouping of some genotypes corresponded with their pedigree information but failed in other cases. The same was true with the geographical origin vis-à-vis grouping of genotypes. Mantel’s test between the cophenetic matrix and the original matrix (z=r=0.923) revealed a good fit of cluster analysis performed. The pattern of grouping in cluster analysis was also reflected in Principal Coordinate Analysis (PCoA) with minor differences. The three coordinate PCoA could account 73.68 per cent of total variability in RAPD markers. The high level of RAPD polymorphism observed in mungbean may be useful for variety protection and monitoring the genetic diversity.ThesisItem Open Access Evaluation of recombinant inbred lines (RILs) of bread wheat for thermo response at seedling stage(CCSHAU, 2008) Sharma, Vikas; Yadava, Ramesh KumarThe present study has been conducted to Evaluation of recombinant inbred lines (RILs) of bread wheat for thermo response at seedling stage. The seed weight (g), seed density (mg ml-3), shoot length (cm), root length (cm), seedling length, fresh weight (mg), dry weight (mg), vigour (I), vigour (II) and germination % in 100-RILs of Harrier x S308 wheat population were observed at three temperature conditions (15º, 20º and 25ºC) with two replications. The estimates of mean, components of genetic parameters (GCV, PCV,h2, genetic advance etc.) and correlation coefficients along with divergence analysis has been worked out after the establishment of substantial amount of variability in the present material. The estimates of h2 (BS) were high for almost for all traits. Genetic advance expressed as % of mean revealed the maximum scope of improvement is for shoot length at 15ºC. Correlation coefficient was indicated to be highest for dry weight and fresh weight at all the three temperatures, and was followed by seedling length with shoot length and that with vigour I at 15ºC; fresh weight with vigour II at 20º and 25ºC and that of dry weight with vigour I at 20ºC and with vigour II at 25ºC. Euclidean Clustering revealed that these RILs could be grouped in 9 clusters at 15ºC, where as in 8 clusters at 20ºC and 25ºC. Only one combination of RILs HS-135 & -136 had the tendency of grouping under same cluster across all the temperatures. Cluster II had maximum xii number of genotypes (24) at 15ºC whereas cluster III at 20ºC, and 25ºC had 20 and 19 genotypes respectively. The cluster VIII had the minimum number of RILs (two) at 15ºC where as four at both 20ºC and 25ºC in each case. At 15ºC RILs in cluster I were better for SL, RL, Se L, VI in cluster III were better for FW, GP, and that of cluster VIII were better for SW, SD, DW, VII as revealed by cluster means, RILs grouped in cluster I at 20ºC were better for RL, Se L, FW, DW, VI, VII; and that of cluster II were better for SW, SL, whereas in case of cluster III were better for SD, GP on the basis of cluster means. Likewise, RILs in cluster I were better for SW, SD; of cluster II being better for FW, DW, VII, GP, and that of cluster III as better for SL, RL, Se L, VI , as indicated by cluster means at 25oC. Consistently higher mean values at all the three temperatures has been recorded in HS 104 for fresh weight, HS 3 for vigour I & HS 51 for germination %, whereas the RIL HS 98 had consistently lower means in case of fresh weight, dry weight and vigour II, HS 99 for shoot length & HS 86 for dry weight at all the three temperatures. RILs specific for improving the traits, shoot length, root length, seedling length, fresh weight and the various seedling vigour parameters of wheat under diverse temperature conditions have been identified.ThesisItem Open Access Genetic and symbiotic characterization of poly-β-hydroxybutyrate production in Rhizobium and Bradyrhizobium spp. nodulating mungbean [Vigna radiata (L.) Wilczek](CCSHAU, 2008) Sandeep Kumar; Yadav, A. S.Twenty-one WT strains of Rhizobium/Bradyrhizobium sp. (Vigna) were isolated purified and tested for their ability to form nodules on the mungbean plants. These WT strains were screened for PHB production on Beringer’s MM with Nile blue A under UV light. The two WT strains MBR 16 and MBR 25 produced high amount of PHB and these were used to isolate PHB mutants. Three types of mutants were isolated and characterized as MHt, MM and ML mutants. The MHt mutants produced highest amount of PHB, whereas ML mutants produced very low amount of PHB. The Maximum amount of PHB was found in MBR 16 MHt 1 i. e. 1.24 g/l. The antibiotic resistance pattern of parent strains and their mutants was almost same. The symbiotic effectivity of parent strains and their mutants on mugbean plants showed that the MHt mutants had higher shoot dry weight and total shoot nitrogen than the MM and ML mutants. A highly significant positive correlation was found between the amount of PHB produced and shoot dry weight and total shoot nitrogen of the munbean plants.ThesisItem Open Access Studies on wheat-azotobacter interaction under pot house conditions(CCSHAU, 2008) Kusum; Vasudava, ManjulaTo study the varietal behaviour of wheat with regard to different strains of Azotobacter chroococcum, as experiment was conducted in which six varieties of wheat such as C-306, PBW-343, WH-283, WH-542, WH-147 and WH-711 were treated with two mutants of A. chroococcum Mac27 and HT54 at three levels of fertilizer such as O kg N ha-1, 90 kg N ha-1, 120 kg N ha-1 in terms of agronomical parameters under pot house conditions. Response of wheat varieties was studied towards A. chroococum in plant height, seed yield, total nitrogen content and total biomass. Variety C-306 showed best response towards both the strains of A. chroococcum. Mac27 showed better response as compared to HT54. Root exudates of these wheat varieties were analyzed for chemotactic behaviour and other components such as sugars, amino acids, phytohormones and organic acids. Glutamic acid and some other unidentified amino acids were detected in both inoculated and uninoculated wheat varieties. Succinic acid was present in wheat varieties WH-711 and WH-147. Maleic acid and some unidentified organic acids were detected in inoculated and uninoculated wheat varieties C-306, PBW-343, WH-283 and WH-542.ThesisItem Open Access Rhizobacterial strains as biocontrol agents against various soil pathogens(CCSHAU, 2008) Chauhan, Sapna; Vasudeva, ManjulaA total of fifty three isolates/mutant were screened for their antagonistic potential against three selected fungal soil pathogens; Rhizoctonia solani cotton, R. solani rice and Fusarium oxysporum. Out of these 10, 6 and 12 isolates/mutant of Azotobacter chroococcum were selected against R. solani cotton, R. solani rice and Fusaraium oxysporum tomato respectively. 19.61% showed zone of inhibition against R. solani cotton, 11.76% against R. solani rice and 23.53% against Fusarium oxysporum tomato respectively. Further quantitative studies were made and these zones of inhibition were measured. In R. solani cotton, the zone was maximum (2276.5 mm2) with AH2 and minimum (141.3 mm2) with AM1. Similarly, in R. solani rice, the zone of inhibition was maximum (1243.44 mm2) with AVK42 and minimum (120.5 mm2) with AR4 and in Fusarium oxysporum, maximum (1004.8 mm2) with HT54 ii and minimum (164.05 mm2) with 103. Further, various studies were done to isolate, evaluate and characterize the antimicrobial substance responsible for this activity. The antimicrobial substance responsible for the antagonistic activity was found to be extracellularly produced and it was not thermostable. TLC assays were done to confirm the production of antifungal and antibiotic compounds taking Pseudomonas as standard. The invitro results were further checked in pothouse conditions on three crops, cotton, guar and tomato respectively. Increase in percent germination was observed. Maximum percentage of disease index (80.36 % and 93.75% on 60th day) was observed with R. solani cotton and R. solani rice check respectively while minimum disease index was seen in case of 103DS (16.67 %) in R. solani cotton and (2.5%) with 103DB in R. solani rice in cotton crop. Similarly in guar crop, maximum percentage of disease index (95.24% and 85.93% on 60th day) was observed with R. solani cotton and R. solani rice check respectively while minimum disease index was seen in case of AH2 (2.5%) in R. solani cotton and (13.29%) with AR4 in R. solani rice. Further in case of tomato, maximum disease index (85 % on 60th day) was observed in control Fusarium oxysporum check and was negligible in strains HT57, AP4, AVK42, AVK51, 103, AH2 and 103DS. The disease was completely controlled by these selected isolates/mutants.ThesisItem Open Access Assessment of genetic diversity in a collection of red rices(CCSHAU, 2009) Chaudhry, Narender; Ahuja, UmaRice (Oryza sativa L.) is one of the important world’s cereal crops, providing staple food for nearly one-half of the global population. It is the only cereal where weedy, wild and cultivated forms are found. In traditional rice growing areas of Asia, in addition to white, rices of red, purple, black, brown, yellow and green colors are known and grown. Rices with red bran layer are called red rices. Some of the wild, cultivated and most of the weedy rices are red. ix Wild species of Oryza are an important source of useful genes. Red rice varieties have tolerance towards stress environments and resistance against important pest, diseases and storage pests. Off type plants having red seeds are considered as weedy although a few of them are colorless. This form of rice is found mixed into cultivated rice fields and shares traits common in both cultivated and wild rice.These are termed ‘Fat beggars’ as they accept what is offered and thrive. These are most difficult to eradicate. They cross with cultivated varieties and produce herbicide tolerant progeny. These have been studied for understanding the evolution of rice and as germplasm tolerant to adverse conditions for rice breeding. At present no cultivated red or wild variety is reported from Haryana though weedy rices are found throughout the state. There is no information available on weedy rice except eradication in Haryana. A collection of cultivated, weedy and wild rices were evaluated for variability at morphological and molecular level.Collection of weedy rices showed significant variability at morphological and molecular level and shared many traits with wild, cultivated white and red rices. Some of weedy rices can be further evaluated for important traits. Diversity at morphological level did not coincide with diversity at molecular level as compared by Euclidean square and UPGMA cluster analysis. 9 primers could be of significance in identification of red rices.One primer i.e Pr836 was found to be unique to weedy rices and giving unique bands in W8 which is long grain like basmati rice, hence can be used to detect adulteration.ThesisItem Open Access Cytological studies of mutagen treated seeds of karanj (Pongamia pinnata L.)(CCSHAU, 2009) Ahlawat, Kavita; Saharan, R.P.The studies involve the treatment of Pongmia pinnata L. Pierre (Karanj) with two mutagens i.e. gamma rays and Ethyl methane sulphonate (EMS) at different concentrations. Three hundred seeds of Karanj were treated for each dose of gamma rays at the dose rate of 10 kR, 20 kR, 30 kR, 40 kR, 50 kR, 60 kR and 70 kR. Similarly three hundred seeds of Karanj were soaked in freshly prepared solution of EMS for each concentration of 0.1%, 0.2%, 0.3%, 0.4%, 0.5% and 0.6% for 6 hours. The treated seeds were sown in the nursery of the Department of Forestry, CCS HAU, Hisar in three replications. The results indicated the effect of both mutagens on the parameters studied. The percent germination decreased as the dose increased in case of both the mutagens. The treatments were significantly differ from each other in case of percent seed germination. LD50 value was found to be 20 kR in case of gamma rays and 0.3% in case of EMS treatment. Root length and plant height decreased with the increase in concentration in case of both the mutagens. The treatments were significantly differ from each other for root length in case of EMS treatment. Collar diameter showed different pattern for both the mutagens. Treatments differ significantly for collar diameter in case of gamma rays and collar diameter decreased as the dose increase, but in case of EMS, the collar diameter increased as concentration increase but at 0.4% show a slight decrease. There was no significant difference among treatments for number of leaves per plant in case of both the mutagens. Mitotic and metaphase indices show an overall decrease with increase in concentration in both the mutagens. The chromosomal abnormalities were increased with an increase in concentrations in case of both the mutagens. Anaphase bridges and distorted metaphase were the main abnormalities found in case of gamma rays and anaphase laggard, anaphase bridge and distorted metaphase were the main abnormalities found in case of EMS treatment. The use of mutagens have been suggested with precautionary measures.ThesisItem Open Access Molecular analyses of recombinant inbred lines of wheat cross aldan x WH542 for resistance to Karnal bunt (Neovossia indica)(CCSHAU, 2010) Gupta, Mamta; Chawla, VeenaPresent studies were conducted on two genotypes, Aldan (resistant) and WH542 (susceptible), and their 94 recombinant inbred lines. Investigations were carried out to screen the recombinant inbred lines (RILs) for Karnal bunt resistance and to characterize above lines using molecular markers. There was significant variation among all the recombinant inbred lines for coefficient of infection of Karnal bunt. The range of coefficient of infection (CI) on the RILs was 0-41.70. The chi-square analysis indicated that a single dominant gene was responsible for Karnal bunt resistance. Thirty five inter simple sequence repeat primers were used for amplification of DNA from the two parents. DNA amplification was observed with only 25 markers. ISSR markers based finger print database were generated using these 25 primers. Out of 25, six primers showed 100% polymorphism. A total of 193 alleles were detected at 25 ISSR loci. The number of alleles per locus ranged from 4-14 with an average of 7.72 alleles per locus. The two parents were genetically divers (similarity coefficient 0.419). The cluster analysis led to distribution into two groups: group 1 had resistant parent Aldan and group 2 had susceptible parent WH542, resistant bulk and susceptible bulk. Among all the primers used one primer (IS-43) was identified to be associated with Karnal bunt resistance. This showed an amplification profile (650 bp product) characteristic of resistant parent, in corresponding bulk and in individual genotyping of the recombinant inbred lines. This might have an application in marker assisted selection after converting it to SCAR marker.ThesisItem Open Access Combining ability analysis for spike traits and other yield attributes in bread wheat (Triticum aestivum L. em Thell)(CCSHAU, 2009) Gollen, Babita; Yadav, R.K.Line x tester analysis involving twelve diverse wheat genotypes (HD2009M, R-6, R-49, HT90, HJP81, SG215, DI717, HS-11, HS-17, HG-2, Rm-Ts13 and HS-27 as lines in combination with six genotypes (WH711, PBW502, Rm-Ts10, Rm-Ts17, C591 and RVG) as testers was conducted by raising the experimental material at the research farm of the Department of Genetics during Rabi 2008. It has been exhibited that PCV is higher than the respective GCV and however, it is slightly lower for most of the characters thus indicating a high magnitude of heritability estimate in broad sense for most of the characters in general. Estimates of combining ability and magnitude of association between various character combinations have been worked out. The results indicated the existence of adequate genetic variation for all the characters studied. Both additive and non-additive type of genetic variance prevailed. C591 and Rm-Ts10 among the testers and HD2009M, HS-17, HG-2, DI717, R-6 and SG215 among the lines were found as good general combiners for one or the other characters. HS-17 for days to heading, HG-2 for plant height and peduncle length, R-6 for 100-grain weight, DI717 for number of grains per spike, SG215 for yield per plant have high GCA effects. Higher SCA effect was observed in the cross HD2009M x PBW502 followed by HS-27 x Rm-Ts10 and HG-2 x WH711 for the character number of grains per spike indicating the preponderance of non-additive gene action in these crosses. WH711 had positive SCA effects in six, PBW502 in nine, Rm-Ts10 in twelve,Rm-Ts17 in nine , RVG in ten and C591in twelve cross combinations. Most of the cross combination involve either both lines and testers or one of the parents as showing good combining ability thus indicating the importance of both additive and non-additive type of gene action. The coefficient for genotypic correlation for most of the character combination was higher in magnitude as compared to corresponding phenotypic correlation coefficient indicating the strong inherent nature of the traits. Grain yield per plant had a strong positive association with days to heading, plant height, peduncle length, tillers per plant, spike biomass and weight of grains per spike by considering both homozygous and heterozygous nature of material. Only few cases of correlation coefficients for the character combinations as per the pattern of heterozygous material has been visualized from the homozygous material which reveals that most traits are under the influence of polygenes and the heterozygous combination for them as has been obtained in various forms depending upon the parents. The informations generated from the present investigation may be utilized by involving good general combiners and the specific crosses that are likely to transgrants for various characters individually or in combination and appropriate breeding approach has been suggested.