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ThesisItem Open Access Anatomical and genetic analysis of damaged grains in rice (Oryza sativa L.)(Punjab Agricultural University, Ludhiana, 2019) Batra, Ritika; Mangat, Gurjit SinghRice grain discoloration or rice black pinhead damage reduces the yield of milled rice and directly reflects on consumer acceptability. The black pinhead damage in grains of PR125 and F3 population of cross PR121 X PR125 and PR121 X PR126 were studied for its percentage damage intensity, anatomical evaluation and marker analysis. The range of percentage damage intensity for both the crosses was 0.5-7.2 and 0.5- 4.9 per cent respectively. The anatomical evaluation was done using scanning electron microscopy (SEM) and the results showed that there was malformation of starch granules at cellular level. The starch granules of damaged grains were loosely packed while they were densely packed in undamaged ones. 200 SSR markers were applied to check parental polymorphism and 15% (30 out of 200) polymorphism was recorded between parents. Two bulks were prepared for bulked segregant analysis, in which one was with maximum percentage damage and other with no damage. From the above 29 polymorphic markers, only 10% polymorphism was observed in the bulks. Chi square analysis was done to check the inheritance of black pin head damage. Since the statistical analysis followed normal distribution, the damage was attributed to be controlled by two or more genes.ThesisItem Restricted Marker assisted selection in wheat for high carotenoid and protein content in grains(Punjab Agricultural University, Ludhiana, 2019) Padhy, Asish Kumar; Sharma, AchlaBiofortification in wheat has been always a neglected facet in the breeding programmes. The research was planned to biofortify the wheat grain with higher carotenoid and protein with low polyphenol oxidase activity and rust resistance. 7E translocated segment containing Y gene/Lr19/Sr25 from Lophopyrum ponticum, GpcB1/Yr36 and Yr15 were the genes to be pyramided in elite agronomic background for high carotenoid content, high grain protein content and stripe rust resistance respectively. 160 lines having Y gene in HD2967 background with the gene in 7A and 7D separately were screened for the presence of Y gene and their carotenoid content and other yield related traits. Biochemical observations showed the differential expression of the gene in different genotypes and increased carotenoid content up to 14ppm where probably, two doses of gene were pyramided in a same genotype (7A and 7D) in F2 population while the commercial hexaploid wheat varieties have carotenoid in the range of 3-4 ppm. UCW8 and BARC8 were used to screen the genes GpcB1 and Yr15 and phenol reaction was carried out for testing the poly phenol oxidase activity. In the F2 population, out of seven crosses synthesized, only one gave good results for carotenoid content while three crosses died in F1 generation due to necrosis. A total of 16 lines were identified which showed high carotenoid and protein content in grains with low PPO activity. The lines having GpcB1 gene were also screened for their yield characteristics and related traits, out of which 3 lines have been identified with high grain protein content and relatively equivalent yield with the commercial checks.ThesisItem Open Access Effect of temperature on germination and alpha-amylase gene expression in wheat (Triticum aestivum L.)(Punjab Agricultural University, Ludhiana, 2019) Gagandeep; Saini, Johar SinghThe present study was conducted with the aim to determine the effect of temperature on alpha-amylase gene expression and germination in wheat. The alpha-amylase gene expression of the chromosome 2 specific gene copies was studied over three temperatures i.e. 10°C, 20°C and 30°C using real time PCR. Differential response of genotypes was observed and KSG 09, KSG220 showed maximum gene expression at 20°C, KSG 30 at 30°C and KSG 072 at 10°C correspondingly. The time period 1-2 days after imbibition was found most crucial for studying the molecular basis underlying alpha-amylase gene expression. Also, rise in gene expression was detected till day 5 and followed by decline upto day 8. Evaluation of forty genotypes for response to germination under varying temperature showed that at 29.2°C, germination percentage was significantly higher than 20.1°C and 14°C however; there was no significant difference in germination percentage between 20.1°C and 14°C. Moreover, interaction between genotype and environment was identified as key aspect for selecting heat tolerant wheat genotypes.666666666666ThesisItem Restricted Genetic Variability and Molecular Characterization for Sucrose Synthesis Gene (s) in sugarcane (Saccharum spp. complex)(Punjab Agricultural University, Ludhiana, 2019) Jagmeet Singh; Thind, K.S.In the present studies, a set of one hundred twelve diverse varieties/clones including twelve checks of sugarcane were evaluated for variability, heritability, genetic advance and correlation coefficient. The aim of present study was to study genetic variability and molecular characterization for sucrose synthesis gene(s) in Sugarcane (Saccharum spp. complex). All the genotypes were planted in a augmented design. The observations on eight cane yield, its components traits and twelve quality traits were recorded. Analysis of variance revealed significant differences among clones/varieties for all the traits. For agromorphological traits, 19 new clones were superior than high yielding check variety CoPb-93.For quality traits, 8 new clones superior than quality check variety CoJ-64.Two clones K-11/9 and K-11/4 were recorded as high yield clones than high yielding check variety CoPb-93 and high quality than high quality check variety CoJ-64.Two clones K-31/2 and K-47/115 better in quality with CoJ-64 and at par the yield with CoPb-93.High magnitude of coefficient of variation i.e. PCV and GCV under cane yield and its component traits was recorded for number of shoots at 120 days and cane yield. High heritability coupled with high genetic advance recorded for number of shoots at 120 & 240 days (000/ha) and cane yield (kg/plot).For quality traits, high PCV, GCV and ECV was recorded for fibre (%). The genetic diversity in a set of 124 (112 clones and 12 check varieties) genotypes of sugarcane was assessed for sucrose phosphate synthetase gene with 10 different set of SSR markers. Different polymorphic percentage that shows high level of diversity was obtained in eight SSR markers. Marker A, SPS, SCB07, MSSCIRI and MSSCIR43 showed higher polymorphic percentage (100%).The divergence analysis grouped 124 genotypes into three major clusters. It was observed that more number of clones belongs to Cluster II. In Cluster III, eight clones and one check (CoJ-88) were present. Maximum similarity was observed in K-1/6 and K-1/30, K-3/18 and K-47/93, K-1/9 and K-48/103 .Minimum similarity was observed in K-11/9 and K-12/13, K-47/41 and CoPb-93. The varieties/clones having high yield and sugar and with high genetic diversity were identified that can be utilized for future evaluation on large scale in sugarcane breeding programme.ThesisItem Restricted Gamma rays irradiation induced variability for confectionery traits and collar rot resistance in groundnut (Arachis hypogaea L.)(Punjab Agricultural University, Ludhiana, 2019) Narinderpal Singh; Brar, K.S.In the present investigation, a set of 35 confectionery groundnut (Arachis hypogaea L.) genotypes was irradiated at 200, 300, 400 and 500 Gy gamma irradiations for standardizing the optimum dose that could be used to irradiate the same set of genotypes for screening for collar rot resistance and obtaining variation in morpho-agronomic characteristics, yield and biochemical components desirable for use of groundnut in food and confections. Optimum dose was obtained on the basis of 50 per cent growth reduction (GR50) in vitro of seedlings raised from irradiated seed by subjecting both, seedling vigour index I and II to probit analysis. Effect of gamma irradiation on growth parameters (germination percentage, shoot and root length, total seedling length, seedling dry weight) was calculated to obtain values for both vigour indices. All these growth parameters were found negatively correlated with gamma irradiation. Optimum dose range inferred was 244-520 Gy. Based on this, same set of genotypes was irradiated at 200, 300, 400 and 500 Gy gamma irradiation to raise M1 generation in vivo to obtain M2 population. Per cent disease incidence (PDI) of all the genotypes increased with increase in gamma radiation in M2 generation. Resistance reaction for any genotype at any gamma ray dose was not observed. Morpho-agronomic characteristics (per cent emergence, plant height, number of primary branches and leaves per plant, leaflet length and width) and maturity period generally decreased and increased respectively, with increase in gamma irradiation in both M1 and M2 generations. This reduction was more pronounced in M1 compared to M2 generation mainly for per cent emergence, plant height and days to maturity indicating restoration of damaged DNA in M2 generation that occurred during M1 generation, while other parameters remained unaffected or showed variation due to environmental heterogeneity. Though, effect of gamma irradiation on yield components (pod number, yield and sound mature kernels per plant, 100 seed weight and shelling percentage) observed a negative shift, some genotypes at different gamma ray doses showed increase in values compared to the control, thus providing desirable variation for yield improvement. Among biochemical components, only sugar content observed a significant reduction, while protein and oil content were not significantly affected by gamma irradiation.ThesisItem Open Access Molecular characterization of introgression for YMV in backcross derivatives of soybean cultivar, JS335(Punjab Agricultural University, Ludhiana, 2019) Koraboyana Vijaya Kumar; Gill, B.S.Yield improvement is the primary objective in soybean breeding. Many biotic and abiotic stresses are responsible for yield reduction of soybean. Among the biotic stresses yellow mosaic virus (YMV) is very serious disease. In India, soybean is prone to 80% yield lose due to YMV. All dominant varieties of soybean are susceptible to this disease. Therefore, the present study was planned to identify the soybean genotypes which are resistant to YMV. JS335 is centrally released high yielding variety but highly susceptible to YMV. Punjab agricultural university developed and released YMV resistant variety SL525. In this study 84 genotypes from four populations viz. F2:7, BC1F6, BC2F5, BC3F4 of the cross JS335xSL525 were screened to identify genotypes having YMV resistance from SL525 and yield and component traits from JS335. The variety JS 335 was used as recurrent parent. The study comprised of three experiments. In first experiment all the genotypes were grown at two locations viz. Ludhiana and Gurdaspur. Analysis of variance of genotypes revealed that genetic variation for grain yield and component traits was present in genotypes. Grain yield was observed to be positively correlated with number of pods per plant, days to maturity and number of seeds per pod. Path coefficient analysis revealed that number of seeds per pod, number of pods per plant and days to maturity recorded with maximum positive direct effect on grain yield. A total of 350 SSR markers were tested for polymorphism on both parents. Out of these, 64 were polymorphic and covering entire genome of 19 linkage groups except LG D1a. By using these 64 polymorphic markers recovery of recurrent parent genome (RPG) was estimated. Graphical genotyping (GGT) was done to identify the areas of introgression and RPG recovery. GGT analysis revealed that maximum RPG was recovered in linkage groups A2 and O. Maximum introgression was occurred in linkage groups C2 and L. Out of 84 genotypes, more than 80 per cent recurrent parent genome was recovered in six genotypes viz. SLJSB-3-3 (86.5%), SLJSB-3-7 (84.4%), SLJSB-3-13 (81.2%), SLJSB-3-10 (81.2%), SLJSB-2-10 (81.2%) and SLJSB-3-14 (80.9%). RPG for each population was also estimated. It was found that as number of back crosses increases, recovery of RPG also increased. Furthermore, phenotypic screening was also done to study the impact of RPG on grain yield and component characters. It was found that grain yield increased in accordance with increase in RPG. Validation experiment was conducted to identify the markers linked to YMV resistance. Out of 7 markers, 3 markers viz. GMAC7L, BARCSOYSSR_02_0547 and Satt322 were successfully validated as linked to YMV resistance. These markers can be used in MAS for YMV resistance.ThesisItem Open Access Response of chicory (Cichorium intybus L.) to variable sowing and harvesting schedule(Punjab Agricultural University, Ludhiana, 2019) Shawinder Singh; Brar, Amandeep SinghInvestigation entitled ―Response of chicory (Cichorium intybus L.) to variable sowing and harvesting schedule‖ was undertaken at Student’s Research Farm, Department of Agronomy, Punjab Agricultural University, Ludhiana during rabi 2016-17 and 2017-18. The soil of experimental field was loamy sand in texture, low in available N and high in available P and available K. The experiment was laid out in split plot design keeping four sowing dates (November 1, November 15, December 1 and December 15) in main plots and four harvesting dates (April 15, April 30, May 15 and May 30) in sub plots. Number of days taken for initiation and completion of emergence were found to increase with delay in the date of sowing. Growth parameters viz; plant height, number of leaves per plant, leaf area and yield attributes viz; root length, root diameter, root volume and root weight per plant were significantly higher with first date of sowing on Nov. 1 than later sowing dates of Nov. 15, Dec. 1 and Dec 15. Among the harvesting dates, growth parameters were not significantly influenced as no treatment was applied at that time , however, at the time of harvesting yield attributes viz; root length, root diameter, root volume and root weight per plant were significantly higher with delayed harvesting date on 30 May. Sowing date of Nov. 1 with harvesting date of April 30 recorded root yield statistically at par with sowing date of Nov. 1 with harvesting dates of May 15 and May 30. Delay in date of sowing from November 1 to November 15 caused significant decrease in growth and yield attributes, resulting into considerable reduction in root yield but this yield reduction can be compensated by delayed harvesting on May15. Delay in sowing caused decrease in TSS and inulin in root as well as NDF, ADF and ash content in foliage, while, crude protein in root and foliage showed increasing trend with delay in sowing. Likewise, delay in harvesting resulted decrease in inulin and crude protein content while TSS showed increasing trend with delayed harvesting.ThesisItem Restricted Inheritance studies of stripe rust, leaf rust and spot blotch diseases in barley (Hordeum vulgare L.)(Punjab Agricultural University, Ludhiana, 2019) Gurinder Pal Singh; Simarjit KaurThe present study was carried out at research farm, Department of Plant Breeding and Genetics, Punjab Agricultural University, Ludhiana, Punjab to study inheritance of stripe rust, leaf rust and spot blotch diseases of barley (Hordeum vulgare L.) in hulless barley genotype PL891, which was found resistant to these foliar diseases since last three years. Chi-square analysis was used to study the number of resistance genes for stripe rust pathotypes; G, M and 24, leaf rust pathotypes; H1, H4 and H5 and spot blotch resistance in PL891. Genetic analysis of F2 generation of PL891 X Jyoti revealed single dominant gene (3 resistant: 1 susceptible) for stripe rust pathotype M and 24 while for pathotype G, F2 showed inhibitory gene action (13 resistant: 3 susceptible). For leaf rust pathotypes H1 and H4, F2 population of PL891 X Jyoti segregated for two independent dominant genes showing duplicate gene action (15 resistant : 1 susceptible) while for pathotype H5, F2 population segregated for inhibitory gene action. Resistance to spot blotch in line PL891 was found to be controlled by single recessive gene. The inheritance studies for these three foliar diseases will be helpful in further gene mapping and transfer of resistance genes into other elite barley cultivars which have become susceptible to these diseases.ThesisItem Restricted Genetic enhancement for kernel elongation of rice variety Basmati 370(Punjab Agricultural University, Ludhiana, 2019) Parminder Singh; Sidhu, NavjotIndia is a major exporter of Basmati rice in the international market due to exemplary quality traits. Attempts were made to characterize the aromatic rices which possess a wide array of diversity which can be incorporated in Basmati improvement programmes. A set of 36 aromatic accessions was evaluated for various morphological and quality traits during Kharif 2017. Significant genotypic variations was observed for days to 50% flowering, plant height, ear bearing tillers/m2, number of grains per panicle, lodging, grain yield, milled rice recovery, head rice recovery, grain length, grain breadth, length/breadth ratio and kernel length after cooking. The genotypes Laldhan, Hung-mi-hriang-matsan, UPR 35565-10-1-1, IET 22187 (RP 4594-121-148-24-11) and Basmati 867 were early in maturity. The genotypes, IR 841- 85-1-1-2 was early maturing, semi-dwarf, and high yielding along with good tillering ability and high number of grains per panicle. Another genotype, IET 22187 (RP 4594-121-148-24- 11) was early maturing and high yielding along with high grain length and good kernel length after cooking. Two genotypes, Bony Cay and Kalikhasa showed resistance to bacterial blight against pathotype PbXo-7. These genotypes can be used in Basmati improvement programmes for various traits. With an aim of combining the elongation ratio of Pusa Basmati 1509 with long slender grain type of Basmati 370, a large F2 population derived from a cross of Pusa Basmati 1509 and Basmati 370 was subjected to quality analysis. Considerable variation was observed for all the progenies. Among these, 53 progenies were identified which possessed desirable grain length suitable for raw milling. Two progenies, 18091 and 18678 also possessed good KLAC and thus can be used to select best segregants in the near future.