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ThesisItem Restricted MOLECULAR CHARACTERIZATION OF Triticum durum – Aegilops speltoides INTROGRESSION LINES AND EVALUATION FOR THERMO–TOLERANCE AND ITS COMPONENTS(PAU Ludhiana, 2013) Zewdu Teshome; Parveen, ChhunejaBackcross introgression lines (BILs) developed from crosses of Triticum durum –Aegilops speltoides were used to identify quantitative trait loci (QTLs) associated with heat tolerance.Three hundred lines in the first season and 100 selected lines in two consecutive cropping seasons were grown under normal sown and late sown conditions. Membrane thermostability, chlorophyll content (CH), canopy temperature and triphenyl tetrazolium chloride (TTC) tests have been used for identifying genetic variability within genotypes.Yield and yield related traits were also recorded. In all the traits variations between BILs in both normal sown and late sown conditions have been observed. Early maturity, shorter height and reduced tillering were recorded in late sown conditions which could be due to terminal heat stress. Out of 386 markers tested, 76 markers (19.7%) showed polymorphism between the parental lines. These polymorphic markers were used to genotype 100 selected BILs. Graphical genotyping showed introgressed regions in all 14 chromosomes of the recipient parent with overall 14.2% introgression.Single marker analysis (SMA) revealed that six markers (Xgwm293, Xgwm565, Xgwm148, Xwmc269, Xgwm299 and Xwmc603) were significantly associated with spikelets/spike at LOD values ranging from 6.1-10.3 and phenotypic contribution of 18.5-44.3%.Similarly, markers Xgwm293,Xwmc269, Xgwm565, Xgwm471, Xgwm148, Xgwm299, Xwmc603, Xwmc31, Xcfd6 and Xcfa2278 were linked to thousand grain weight. One marker Xcfd60 revealed association to TTC in normal sown BILs with LOD value of 3.1 and PVE of 16.1%. Four markers Xgwm293,Xgwm148, Xcfd60 and Xwmc269 showed linkage with CH with varying LOD score and PVE%. SMA also identified eight markers (Xgwm293, Xgwm565, Xgwm471, Xgwm148, Xgwm299, Xwmc603, Xcfd6 and Xwmc269) linked to days to flowering and four markers viz, Xgwm293, Xgwm471, Xgwm299 and Xwmc603 showed association to days to maturity with LOD value higher than the threshold. Plant height was also linked to four marker loci (Xgwm471, Xgwm148, Xgwm565 and Xwmc603) in average environment and stay green trait was linked to Xgwm148, Xwmc269, Xgwm471 and Xwmc269 with LOD score in the range of2.6 -3.4 and PVE of 12.6-15.9%. Additive QTL analysis identified that most of the traits were contributed from the donor parent and a number of markers identified in SMA were validated in additive QTL analysis also.ThesisItem Open Access Effect of alien cytoplasms on heterosis and combining ability of yield, quality and water use efficiency traits in sunflower (Helianthus annuus L.)(PAU, 2013) Tyagi, Vikrant; Dhillon, S KThe present investigation was undertaken in the Department of Plant Breeding & Genetics, Punjab Agricultural University, Ludhiana during the years 2011 and 2012 to study the effect of alien cytoplasms on heterosis and combining ability of yield, quality and water use efficiency traits in sunflower (Helianthus annuus L). Analysis of variance for morphological, physiological and quality traits revealed significant differences due to genotypes and environments (irrigation regimes) for all the characters. Among sources E002-91A recorded a substantial increase in plant height, head diameter, seed yield and harvest index under stress environment, indicating its suitability for developing water use efficient hybrids. The D 2 analysis revealed a lot of diversity between cms analogues. Euclidean distance values indicated the uniqueness of ARG-3A under stress environment. For seed yield and its component traits E002-91A, ARG-3A, ARG-6A and RCR-8297 were good combiners under both the environments while PRUN-29A and CMS-XA under stress environment. The role of female parent was more in expression of all the traits as compared to male parents. The hybrid s CMS-XA x P100R, ARG-2A x P100R, ARG-6A x P69R, DV-10A x P100R, PRUN-29A x RCR-8297, 234A x P69R and 38A x P124R were identified as best specific combiners for seed yield and E002-91A x P124R, E002-91A x P100R, ARG-3A x RCR-8297, ARG-6A x P124R, DV-10A x P69R, PRUN-29A x P100R, 40A x RCR-8297, 40A x P124R for oil content. For seed yield maximum standard heterosis over check PSH-996 was recorded as 24.72 per cent for 40A x P69R (under normal environment) and 60.7 per cent for PRUN -29A x P69R (under stress environment). It is pertinent to mention here that all the sources significantly differed from PET-1 source with respect to seed yield per plant under both the environments and this effect was in positive direction. The sources ARG-3A and DV-10A had significant effect on heterosis for 100 seed weight under stress environment. Similarly for seed yield none of the sources recorded significant effect on heterosis under normal environment while in stress environment all the sources except ARG-2A and ARG-6A had positive effect on heterosis. Seed yield recorded significant positive association with plant height (0.301), highly significant correlation with leaf water potential (0.417) and biological yield (0.372) under normal environment. A total of 11 hybrids giving stable performance over normal and stress environments have been identified for seed yield. The genotypes with stay green trait at maturity were identified under both the environments. Drought susceptibility index (DSI) studies revealed that few genotypes recorded stable performance under both the environments with respect to seed yield having DSI values near to zero.ThesisItem Open Access Molecular marker and morphological characterization of triticale x wheat derivatives(PAU, 2015) Feltaous, Yousef Mohsen; Bains, N.S.The study is based on near homozygous derivatives from three crosses: TL 2908/2*CSS//PBW 343; TL 2908/2*CSS//DBW 17 and TL 2908/2*CSS//PBW 550 where TL 2908 is an improved, amber grained triticale variety, CSS is a bread stock carrying Ph suppressor genes in Chinese Spring background and PBW 343, DBW 17 and PBW 550 are bread wheat varieties used as recipient parents. As all the recipient wheat varieties carried 1B/1R translocation, it was expected to be present in half of the derivatives. Out of 1244 lines screened using SSR marker diagnostic of 1B/1R translocation (SCM 9), 652 turned out to be non 1B/1R. The lines negative for 1B/1R translocation were screened for presence of rye chromatin using a universal marker (Rye R3/F3). A large proportion (266 out of 552 lines, 48.2%) was positive for rye chromatin. A set of 164 derivatives positive for rye chromosome introgression were taken up for evaluation in replicated trials. Transfer of large anther size and high anther extrusion was indicated in several derivatives, which is relevant for hybrid wheat development. Sixty five derivatives showing almost complete resistance to highly virulent stripe rust race 78S84 were evaluated in a separate trial. While all the recipient bread wheat parents were highly susceptible to stripe rust and Karnal bunt, twelve derivative lines combined high level of resistance to stripe rust (mostly 0 or ts) and Karnal bunt (less than 2% infection). Another three lines were found to be highly tolerant to micronutrient deficiency. SSR marker based screening of elite derivatives revealed three lines to carry rye chromosome 5 (and three lines with rye chromosome 6. One line carried both chromosomes 5R and 6R.ThesisItem Open Access Molecular characterization of inter-specific backcross inbred lines of rice (Oryza sativa L.) for mapping of yield component QTL(PAU, 2015) Bhatia, Dharminder; Singh, KuldeepWe need to significantly increase the yield potential of rice for feeding estimated 9 billion populations by 2050. For increasing yield potential in rice, utilization of wild species is one among the several strategies advocated. In the present study, Backcross inbred lines (BILs) derived from O. longistaminata acc. IRGC104301 (longi-BILs), O. rufipogon acc. IRGC104433 (rufi-BILs) and O. glumaepatula acc. IRGC104387 (glumae-BILs) in the background of O. sativa ssp. indica cv. PR114 were used for mapping QTLs consistently contributing variation for yield and yield component traits. BILs were evaluated for yield and yield component traits as compared to recurrent parent PR114 as check in alpha lattice design over three seasons spanning two locations. A modified Genotyping by sequencing approach was used for genotyping. Sequence data for each population was analysed with custom designed method exactly similar for each population to identify SNPs. A total of 3322 informative SNPs of rufi-BILs and 3437 informative SNPs of glumae-BILs were used for mapping QTLs for twelve yield and yield component traits using inclusive composite interval mapping. SNPs identified in longi-BILs could not be used for mapping due to large number of missing data points. In rufi-BILs, QTL for thousand grain weight (qtgw5.1) was mapped on chromosome 5 consistent over all the three seasons and with positive additive effect contributed by O. rufipogon allele. Two QTLs for grain width (qgw5.1 and qgw5.2) were also mapped on chromosome 5 with positive and negative additive effect respectively. In glumae-BILs, three consistent QTL for thousand grain weight on chromosome 2 (qtgw2.1), 3 (qtgw3.1) and 6 (qtgw6.1) were mapped consistently for season 2 and 3. QTLs qtgw2.1 and qtgw6.1were associated with positive additive effect, while qtgw3.1 with negative additive effect contributed by O. glumaepatula allele. Two QTL for grain length (qgl7.1 & qgl7.2) with positive additive effect contributed by PR114 allele was mapped on chromosome 7 spanning overlapping position in both rufi-BILs and glumae-BILs respectively. qgl7.1 and qgl7.2 might be same QTL as contributed by same parent and spanning overlapping position. QTL for other yield and yield component traits could not be identified due to skewed segregation.Yield component QTLs identified in the present study from low yielding wild relatives of rice reveals their significance in improving yield of cultivated rice. Combining these QTLs in the background of cultivated rice will not only increase the yield, but also widen the genetic base of cultivated rice.ThesisItem Open Access Genetic Diversity and Association Studies for traits Related to Water Stress Tolerance in a Structured Brassica Juncea Population(PAU, 2014) Akhatar, Javed; Banga, S.S.A core subset of genetically fixed lines (48) was selected through stratified sampling to represent the allelic variations within Brassica juncea gene pool. The molecular characterization by 158 nuclear and nine chloroplast SSR markers revealed distinct germplasm patterns. There were significant interactions between chlorotype and seed/biological yields. Test genotypes were also evaluated for twenty four morpho-physiological traits over two years under normal and restricted moisture regimes for association studies. Highly significant marker-trait associations (MTA‟s) were identified by General Linear Model [GLM-PC (Principal Component)] Mixed Linear Model [MLM-PC+K (Kinship)]. MTA‟s include days to maturity, grain yield, number of secondary branches, number of seeds per pod, thousand seed weight, average root diameter and root length. Single marker, SB1822-1, localized at 17.5cM on chromosome 13 was repeatedly detected for seed size and grain yield. Marker, SB3872-3 revealed significant effect under normal moisture on seed size during the year one. Among the favorable double alleles, SB1822-1 showed positive phenotypic effect for thousand seed weight and grain yield. Of the four double alleles, cnu316-3 had maximum positive phenotypic effects on grain yield. Maturity QTLs (Quantitative Trait Loci) detected on J6 (A genome) have been repeatedly detected in past studies with diverse in B. napus. Novel QTLs were identified for photosynthetic active radiation interception (cnu031-1 and SB1728-3) and drought tolerance index (NI3-G07-1 and SB1935A-1) and canopy air temperature difference. The loci identified by us need to be verified and utilized for breeding tolerant cultivars to improve crop performance under variable moisture conditions.ThesisItem Restricted Mapping of quantitative trait loci for resistance to ascochyta blight, botrytis grey mould and fusarium wilt in chickpea(PAU, 2012) Garg, Tosh; Sarvjeet Singh, Sarvjeet SinghThe present investigation was carried out using 188 RI lines (F8) developed by crossing two enotypes JG 62 and ICCV 05530. The parent JG 62 was susceptible to three economically important diseases viz., ascochyta blight, botrytis grey mould and fusarium wilt whereas parent ICCV 05530 was resistant to these diseases. The study on genetics of resistance to ascochyta blight showed the monogenic control at adult plant stage at Ludhiana and Dhaulakuan, monogenic and trigenic control at seedling stage at Ludhiana and Hyderabad, respectively. The genetics of resistance to fusarium wilt showed the monogenic and digenic control against the race 3 and race 1 of Fusarium oxysporum. Resistance to botrytis grey mould was digenically controlled at seeding as well as adult plant stage. Linkage map was developed by using JoinMap version 4 with the help of 125 molecular markers and one phenotypic marker which gave total 11 linkage groups spanning a length of 399.6 cM. The major QTL associated with the ascochyta blight resistance at seedling as well as adult plant stage against the isolate 8 of race (3968) was reported on LG01B which estimated 40.87% and 51.40% phenotypic variance, respectively. The QTL associated with ascochyta blight resistance against pathotye I at adult plant stage was also identified on LG01B but QTL associated at seedling stage against Hisar race was identified on LG04B. The fusarium wilt resistance gene against race 3 was present on LG02 flanked by the markers TR19 and H2B061, TA27. Wilt resistance gene against race 1 was also flanked by same markers on G02. One QTL associated with BGM resistance against isolate 4 of race 3 was identified on LG06. In this population, total 38 QTLs were identified for 12 morphological characters. Double podding gene was linked with marker TR44 at 2.5 cM distance on LG06. The maximum number of QTLs associated with morphological characters were identified on linkage group 4. The QTLs/genes identified for important morphological traits as well diseases would be useful for marker assisted selection in the future breeding programmes.ThesisItem Open Access Improvement of rice variety PAU 201 for aleurone colour and bacterial blight resistance using Marker Assisted Selection(PAU, 2013) Prem Kumar A; Gill, M.S.In Punjab, Rice variety PAU 201 was released for cultivation in the year 2007. Within two years this variety occupied 25% of total area under rice because of its high yield. It gives good yield under average conditions. However, recently this variety faced problems because of red aleurone colour, susceptibility to new evolved pathotypes of bacterial blight and low amylose content. During 2010, crosses were attempted between PAU 201 and RYT 3148 to improve the characters of PAU 201 with the help of molecular markers. The F1 plants were backcrossed with recipient parent and BC 1 F 1 seeds were generated. In 2011, marker-assisted foreground selection for first gene rc7 was analyzed on 270 BC 1 F 1 plants, 97 plants showed positive result for this gene in heterozygous condition. For second target gene (Xa21), 23 plants were found to be heterozygous for Xa21 and the desirable plants were backcrossed to recurrent to generate BC2 F 1 seeds. For third gene (wx), 10 plants out of 23 were recorded for the presence of three genes (rc7, Xa21 and wx) while 13 plants had two target genes (rc7 and Xa21). BC2 F 1 seeds were sown in kharif 2012, 371 out of 669 plants were heterozygous for rc7 gene. Among 371 BC 2 F 1 plants, 231 plants were selected which looked similar to PAU 201 in terms of agro-morphological characters. Out of these, 80 plants were found positive for Xa21 and 25 plants out of 80 plants were identified to be heterozygous for RM190. So, on the basis of analysis of three genes, 25 plants were identified to the presence of the target genes. In case of two gene category, 118 plants out of 778 were selected based on morphological characters which resemble to PAU 201. During parental polymorphic survey, 69 polymorphic markers (19.8%) were identified out of 348 between parents. Four and six polymorphic markers in carrier chromosomes 7 and 11 were applied on selected plants to find out the recovery percentage of recurrent parent. The recovery percentage of selected plants varied from 50-75% in carrier chromosomes 7 and 11. Based on GGT results, some plants were observed as heterozygous condition with retain segments from the recurrent parent, while heterozygosity was observed at all loci in the remaining plants for carrier chromosomes 7 and 11. Plants (24-12-43, 24-15-5, 24-18-8, 24-18-44, 24-78-54, 24-78-58, 24-200-17,24-200-18, 24-200-19, 24-200-24, 24-200-47, 24-200-69, 24-200-70, 24-200-71 and 24-200-75) carried three target genes with recurrent parent genome to the extent of 75.0% in carrier chromosomes along with grain size and shape similar to parentsThesisItem Restricted EPIDEMIOLOGY AND MANAGEMENT OF ALTERNARIA LEAF SPOT OF Aloe barbadensis(2009) Pawan Kumar Amrate; J. R. SharmaSurveys conducted during 2008 revealed that Alternaria leaf spot disease of Aloe barbadensis was prevalent at all the locations viz. Ludhiana, Malerkotla, Nawanshahar (Ballowal Saunkhri) and Bathinda. Disease incidence and severity was highest at P.A.U. Regional Research Station, Bathinda which recorded 90.0 and 34.0 per cent respectively. Alternaria alternata (Fr.) Keissler was isolated from diseased leaves and pathogenicity was proved on healthy leaves. Disease development and progression were correlated with prevailing environmental factors at Ludhiana during the year 2008. Highest disease incidence and intensity was recorded during August. However correlation study showed that all the factors (atmospheric temperature, relative humidity and cumulative rainfall) were positively correlated with disease development. This investigation revealed that the atmospheric temperature between 23 to 300C, relative humidity between 65 to 95 per cent and higher rainfall were conducive for development and progression of the disease. To find out effective strategies for the management of the disease various fungicides, plant oils and plant extracts were evaluated in vitro as well as in vivo for their efficacy against the pathogen A. alternata. Among these Amistar, garlic oil and turmeric extract were found to be most effective in inhibiting the spore germination of A. alternata and recorded ED50 values of 5.5 g/ml, 155 g/ml and <10000 g/ml respectively whereas Score was found to most effective in checking the mycelial growth of the pathogen as it completely inhibited the growth even at 25 g/ml and recorded ED50 value of 1.7 g/ml. Garlic oil and turmeric extract were also found to be effective in inhibiting the mycelial growth of the pathogen. In the field experiment, all the tested fungicides reduced disease intensity significantly. Among these Score 25 EC was found to be highly effective as it reduced 80.6 per cent disease at 0.1 per cent concentration whereas garlic oil and turmeric extract also showed promising result. In vitro screening of genotypes for their resistance/susceptibility response revealed that Aloe barbadensis contracted maximum disease and showed 100 per cent disease incidence.ThesisItem Restricted Genetic enhancement of grain zinc content in high yielding wheat varieties(Punjab Agricultural University, Ludhiana, 2016) Charan Singh; Sohu, V.S.Two approaches for genetic enhancement of grain zinc content in high yielding wheat lines was pursued under the study. In the first approach high grain micronutrient content from Triticum monococcum and Triticum boeoticum introgressed into tetraploid (T. durum) background served as donors. The recipient parents utilized were hexaploid high yielding wheat varieties of North Western Plane Zone such as DPW621-50, HD2967 and WH1105 along with two gene pyramided version of PBW343 (carrying rust resistance genes Yr10, Yr15, Lr24, and Lr28). A total 1077 progenies (BC2F2) from different cross combinations were generated and evaluated for grain zinc content other agronomic and quality traits. Analysis of variance for augmented block design revealed that genotypes differed significantly for grain zinc concentration (ppm). A set of highly promising 271 progenies with more than 70 ppm grain zinc concentration and 25 progenies with more than 80 ppm grain zinc concentration were identified. The highest grain zinc and iron concentration was observed in progenies of gene pyramided parent PBW698 with introgression from BF22 used as tetraploid donor. The monitoring of previously mapped grain micronutrient QTL in this cross showed the markers identified at diploid level to be non polymorphic. The second approach followed in this study employed grain micronutrient enhancement via positive association of grain micronutrient and protein content with Gpc-B1 gene, originally derived from T. dicoccoides. A back cross RIL population of 94 Gpc-B1 positive lines derived from cross Glupro/3*PBW568 showing wide variation in protein and micronutrient content and several agronomic traits including plant height and thousand grain weight in a two year evaluation experiment was used in a correlation analysis. Analysis of variation and correlation studies showed that the lines which possess high grain protein content, grain zinc and iron concentration were low yielding with less number of grains per spike, bolder grains, earlier maturity and relatively more plant height. It showed that increase in grain protein content was constrained and a compromise of yield and yield components was observed. Further, yield reduction could arise in more than one ways. For instance, there could be reduction in grain per spike as observed in this study or thousand grain weight as indicated by several reports in the literature. In spite of yield-quality trade off commercially useful product-specific lines can be derived. The relatively tall and early but bold grains lines with exceptionally high protein (15-16%) and micronutrient content (70-80 ppm) in the above studied set can be shortlisted as product specific (bread making) high premium cultivar after further testing. In a second set of experiments under this approach, DPW621-50 derivatives with Gpc-B1 gene were evaluated for yield, yield components and grain zinc content. Pooled analysis of variance and correlation coefficient analysis indicated that Gpc-B1 gene was less effective in high yielding DPW621-50 background. This gene moderately enhanced grain protein content and grain zinc and iron concentration in DPW621-50 derivatives with some reduction in thousand grain weight and grain yield. It was seen that the Gpc-B1 gene must be allowed sufficient flexibility of productivity trait combinations for its protein and grain enhancement effects to be manifested.