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ThesisItem Open Access Evaluation of flag leaf characteristics in relation to spike quality and corm production in gladiolus (Gladiolus grandiflorus L.)(Punjab Agricultural University, 2024) Ramandeep Kaur; Dr. Shalini JhanjiThe present investigation entitled “Evaluation of flag leaf characteristics in relation to spike quality and corm production in gladiolus (Gladiolus grandiflorus L.) was conducted at the Research Farm of the Department of Floriculture and Landscaping, Punjab Agricultural University, Ludhiana during 2022-2023. The genotypes/varieties were evaluated for distinctiveness, uniformity and stability (DUS). Significant differences among the genotypes were recorded for studied morphological and post harvest quality parameters. The cluster analysis grouped the genotypes/varieties into two major clusters which were further subdivided into two subclusters each. Cluster Ib (Punjab Pink Elegance, Punjab Lemon Delight, Punjab Glance, etc) performed best in terms of desirable morphological traits while cluster IIa (Pusa Gunjan, Pusa Kiran, Punjab Dawn, etc) was found to have performed least in terms of various morphological characters. Cluster analysis also revealed that Punjab Glance and Punjab Glad 3 lied in different clusters and differed based on flag leaf length, spike length, spike weight, corm weight, days to initiation and complete spike emergence due to which these two varieties were selected for evaluation of flag leaf characteristics in relation to spike and corm production during different spike developmental stages. With advancement in developmental stages, there was significant growth of flag leaf, spike and corm. In comparison of Punjab Glance and Punjab Glad 3, Punjab Glad 3 had comparatively greater flag leaf length, width, area, weight, total leaf weight, area, spike length, rachis length, spike weight, rachis weight, spike diameter, lamina thickness, epidermal thickness, mesophyll thickness and stomatal index but in terms of corm traits, Punjab Glance had greater number of corms, corm weight, corm diameter and number of cormels. Punjab Glad 3 had comparatively greater membrane stability (MS), relative water content (RWC), total soluble sugars (TSS), total soluble proteins (TSP), sucrose content, starch content, activity of acid invertase, neutral invertase and sucrose synthase than Punjab Glance in both flag leaf and spike while in corm Punjab Glance had greater MS, RWC, TSS, TSP, sucrose content, starch content, activity of acid invertase, neutral invertase and sucrose synthase. Punjab Glad 3 had greater flag leaf length and width which led to longer spike but less corm development as maximum assimilates were translocated to spike while in case of Punjab Glance though it had shorter spike but the corm in this case had greater sink strength which led to better corm development.ThesisItem Open Access Biochemical defense response and genetic basis of resistance against Ascochyta blight (Ascochyta rabiei (Pass.) Labr.) in chickpea (Cicer arietinum L.)(Punjab Agricultural University, 2024) Abassy, Omer; Dr Upasana RaniChickpea (Cicer arietinum L.), the second-largest global pulse crop, is under threat from highly variable nature of Ascochyta rabiei, causing widespread disease damage under favourable environmental conditions. This disease, Ascochyta blight is a major concern worldwide, with insufficient resistant sources and frequent breakdowns due to rapid pathogen evolution. This study evaluated 238 kabuli chickpea genotypes for resistance to Ascochyta blight (AB) over the growing seasons from 2019-2022 at Punjab Agricultural University under artificial epiphytotic conditions, identifying 18 resistant lines (GLK 10-40, GLK 20054, GLK 20055, FLIP 10-298C-IFC-S2, FLIP 10298C-IFC-S1, CS-3-E-24, FLIP 09 256C-55, FLIP 07-314C-57, FLIP 10-243C, FLIP 09-194C, FLIP 08-104C, FLIP 04-219C, ICCV 55233, ICCV 55215, ICCV 55135, ICCV 55108 and ICCV 155141) belonging to different genetic backgrounds with disease scores of 1.0 to 3.0, whereas seventy-three (73) lines demonstrated a moderate level of disease resistance with a disease rating of 3.1-5.0. The activities of defense-related enzymes (PAL, TAL, PPO, POD) associated with phenol metabolism along with lignin and total phenol content were compared among six kabuli chickpea lines, viz., five resistant (GLK 10-40, GLK 20055, FLIP-09-194C, FLIP-04-219C, ICCV 55215) and one susceptible (GLK 17301) treated lines that exhibited differential responses to Ascochyta blight at tested time intervals of 48, 96, 144 and 240 hr post-inoculation respectively. The perusal of data showed increased activity of all enzymes (PAL, TAL, PPO, POD) till 96 hr after inoculation, whereas the non-enzymatic contents such as lignin and total phenols showed the maximum enzyme activity till 144 hr after inoculation. However, enzyme and non-enzymatic activity remained almost constant with no significant difference in all control/untreated resistant and susceptible line(s) at different time intervals. For understanding the genetic basis of resistance, the F2 population comprising of 162 plants derived from the cross, GLK 24096 (resistant) and L 556 (susceptible) was assessed under artificially epiphytotic conditions against local isolate of Ascochyta rabiei. Of 162 plants evaluated, 39 plants were grouped in resistant (R ) category (disease score ≤ 5.0 ) and 123 were classified as susceptible (S) with score of five and above. Consequently, the F2 plants exhibited a segregation ratio of 1R:3S, suggesting that a monogenic recessive gene governed resistance to Ascochyta blight. Of thirty four SSR markers associated with Quantitative Trait Loci (QTLs)/genes were used for confirmation of resistance in identified eighteen resistant and six susceptible lines, only 19 SSR markers depicted polymorphism. Notably, SSR markers, STMS11, TA194, TS12, TA146, and TA2 exhibiting significant polymorphism, indicated tight linkage to Ascochyta blight resistance. Phylogenetic analysis showcased distinct clustering based on resistance, offering insights into the genetic basis of resistance. These findings informed about targeted resistance breeding and sustainable disease management with defence-related enzymes/identified markers holding promise for developing superior, resistant and resilient chickpea varieties contributing to crop sustainability and productivity.ThesisItem Restricted Response of soil and foliar applied zinc on wheat and its residual effect on moongbean(Punjab Agricultural University, 2023) Manjot Singh; Dr B. K. YadavMicronutrient deficiencies have rapidly emerged as a significant human health concern, particularly in developing nations. Zinc (Zn) is a basic ingredient and regulatory co-factor in enzymes and proteins involved in many metabolic activities. It is vital for both animals and plants. The experiment was conducted during the Rabi 2020-21 and Kharif 2021to study the response of soil and foliar applied zinc on wheat and its residual effect on moongbean. Soil and plant samples were analysed by standard methods. The present study results revealed that plant growth parameters such as plant height, spike length, test weight, grain and straw yield of wheat significantly differ among wheat cultivars and increased by spraying of 0.5% ZnSO4.7H2O along with 1% urea at the time of booting + grain filling stages. Macronutrients (N, P, and K) and micronutrients (Fe, Cu, Zn and Mn) content in wheat grains significantly varied among wheat cultivars. The macronutrients (N, P and K) and micronutrients (Fe, Cu and Mn) content in wheat grain and straw was non- significant due to spraying of Zn fertilizer irrespective of the source, however, Zn content in grain and straw varied significantly due to spraying of Zn fertilizer irrespective of the source. The soil chemical properties (pH, EC and OC), macronutrients (N, P and K) and micronutrients (Fe, Cu and Mn) showed nonsignificance and soil available Zn showed significance difference between two cultivars (PBW 725 and PBW 658), under different Zn treatments and time of spray. The residual effect of different Zn sources and time of application on soil chemical properties and available nutrients were also non-significant after succeeding moongbean crop. However, available Zn was significantly higher with applied Zn @ 5kg ha-1 in soil (T1). Similarly, the plant growth parameters and Zn content in succeeding moongbean was significantly higher with applied Zn @ 5kg ha-1 in soil (T1) as compared to other methods of Zn application. From the present study it is concluded that wheat variety PBW 725 showed the highest yield attributes, yield, Zn content and among all the Zn treatments, treatment T5 (2.5 kg Zn ha-1 applied in soil + 0.5% EDTA- Zn +1% urea - foliar spray) exhibited superior performance. Whereas, treatment T1 (5 kg Zn ha-1 -applied in soil) has the most beneficial residual effect on succeeding crops (moongbean) in terms of growth and yield.ThesisItem Embargo CRISPR/Cas9-mediated pectate lyase gene editing for enhanced shelf life of tomato (Solanum lycopersicum L.)(Punjab Agricultural University, 2024) Sumedha; Dr Prashant MohanpuriaTomato is one of the most consumed vegetable crops worldwide. Tomato fruits are having an inherently short shelf life which is directly related to fruit softening, caused by cell wall modifying enzymes. Among which pectate lyase (PL) controls fruit softening and thus PL gene was targeted using CRISPR/Cas9 to enhance shelf life of tomato variety Punjab Ratta. In silico study of PL genes from Solanaceous crops including tomato was attempted and three sgRNAs were designed from exon-2 of PL gene. The CRISPR/Cas9 constructs containing sgRNA-1 and 2 were used for transformation of tomato. A new in planta transformation protocol was developed with 10.49 % overall transformation efficiency. Floral buds of 3-8 days of developmental stages were used for dipping in Agro suspension which resulted in setting of 150 fruits. The extracted T0 seeds were selected on kanamycin. A total of 3240 T1 generation plants were regenerated from transgenic tomato fruits (T0), out of which 1620 were found kanamycin resistant which were used for DNA isolation and PCR confirmation of T-DNA integration. The 425 bp PL gene target region from 170 transgenic tomato T1 plants was PCR amplified. Upon sequencing eight T1 tomato transgenic plants were found with PL gene edits containing T insertion, T insertions as well as T substitutions, GC substitution by CG as well as TT insertion downstream to PAM site. PL gene expression and pectin content were decreased in fruits of mutated tomato plants but morphological and biochemical characteristics were more or less similar.ThesisItem Embargo Characterization of maize inbred lines for high plant density amenable traits(Punjab Agricultural University, 2024) Meena, Tarun Kumar; Dr Gagandeep SinghIn temperate maize, breeding for stress tolerance including a higher number of plants per unit area has been extensively used for productivity enhancement in commercial hybrids but this strategy has not been exploited in tropical maize. The increase in the number of plants per unit area through the adoption of high density planting (HDP) is one of the alternative strategies to achieve high productivity in tropical/subtropical maize. Narrow leaf angle, shorter anthesis silking interval, and moderate plant height were delineated as major architectural traits for tolerance to HDP in parental lines. The lines harboring these traits coupled with high grain yield potential under HDP were identified and recommended to use as parents for the HDP breeding program. These are following inbred lines identified for HPD traits CML 451, 221520, EML108, EML113, EML129, PML1112, PML115, PML1194, PML1239, PML133, PML162, PML163, PML164, PML18, PML2, PML80, PML81, SN485145 and VL174449.The inheritance study of leaf angle resulted as the expected number of intermediate, broad leaf angle and narrow leaf angle plants for 9:6:1 was 405, 250 and 607, respectively, giving a chi-square value of 1.097 (< 5.991) hence giving a perfect fit to 9:6:1. Thus, Chi-square analysis revealed that the narrow leaf angle is controlled by two recessive gene. A cross between broad leaf angle (PML1126) and narrow leaf angle (CML451) inbred line was used to raise an F2 mapping population for the construction of two contrasting bulks namely, broad leaf angle (BLA) and narrow leaf angle (NLA). These bulks along with both the parents were sequenced using whole genome resequencing and analysed using the SNP-index method for identifying the narrow leaf angle associated region. Identification of QTLs associated with narrow leaf angle in the material under study based on sequence based bulked segregant analysis and use of QTLseqr software was done. It was observed that the ∆SNP-index values in the region 48.72mb to 55.13mb on chr 6, 44.63mb to 45.74mb; 46.21mb to 50.86mb on chromosome 7, 48.22mb to 54.56mb on chromosome 9 lied outside the statistically calculated confidence interval at 99% level of significance. The findings of the present study will facilitate in genetic dissection of candidate gene(s) responsible for the narrow leaf angle gene. This study will also help in the development of SNP-based markers which would facilitate marker-assisted transfer of narrow leaf angle gene in maize inbred line.ThesisItem Restricted Genetic and biochemical characterization among diverse set of Linseed (Linum usitatissimum L.) germplasm(Punjab Agricultural University, 2024) Alka; Dr. Arvinder Kaur ToorThe present study involved the evaluation of sixty linseed genotypes at two locations. Two experiments were conducted, focusing on agro-morphological and biochemical traits. The agromorphological assessment included traits like branches, capsules, seed yield, and maturity duration. Genotype BAU-2019-13 excelled in seed yield at Ludhiana, while LCK-2134 performed best at Gurdaspur. Descriptive statistics indicated minimal environmental impact on most of the traits, suggesting potential for improvement through direct phenotypic selection. Traits like primary branches and seed yield per plant showed high heritability and genetic advance, indicating suitability for improvement through phenotypic selection. Genotype × environment interaction was observed, and GGE biplot analysis highlighted superior performances of LCK-2134 in Gurdaspur and BAU-2019-13 in Ludhiana. Positive correlations were identified between morphological traits and seed yield per plot. Dendrograms revealed six clusters at each location, by grouping genotypes with similar features. These clusters are crucial for breeding programs and for suggesting potential for heterosis. The agro-morphological characterization demonstrated significant genotype variation. The second experiment focused on biochemical traits, including crude protein, oil percentage, and fatty acids. Genotypes with high oil percentage and low anti-nutritional factors were identified as valuable for both human and livestock feeding. The top genotypes for oil percentage and crude protein were Surbhi × LC 2023 and BAU-2021-08, respectively. Genetic variability was confirmed by significant mean sum of squares in the analysis of variance. G × E interaction was significant for tannins and crude protein, while other traits showed non-significant interaction. Dendrograms for biochemical traits classified genotypes based on homogeneity and heterogeneity. The study's comprehensive characterization identified promising genotypes with exceptional performance in both agro-morphological and biochemical traits. This information is valuable for future linseed breeding programs.ThesisItem Embargo Effect of different paired row arrangement on growth, yield and quality of kharif maize (Zea mays L.)(Punjab Agricultural University, 2024) Ashesh, Ayush; Dr. Harpreet SinghThe present study entitled “Effect of different paired row arrangement on growth, yield and quality of kharif maize (Zea mays L.)” was conducted at Regional Research Station (RRS), Gurdaspur and Punjab Agricultural University (PAU), Ludhiana during kharif 2022. The soil of Regional Research Station, Gurdaspur and Punjab Agricultural University, Ludhiana was clayey loam and sandy loam respectively and pH of soil at both the locations is neutral. The research was conducted in Randomized complete block design having twelve treatments and three replications. The hybrid variety used during research was PMH-13. The treatment comprise of 9 paired rows and 3 conventional methods at standard spacing of 60 cm x 20 cm. The paired rows are at different spacing where intra-paired rows are 20, 30 and 40 cm with inter-paired row distance of 60, 80 and 100 cm. Plant height significantly varied with planting population and paired spacing. The treatment with 20 cm intra-row spacing and 60 cm inter-row spacing consistently resulted in taller plants at various stages of growth. The treatment with 40 cm intra-row spacing and 100 cm inter-row spacing, which had the lowest population, exhibited significantly thicker stem girth. Days to reach tasseling and silking stages were not affected by different paired row arrangements and population densities. Leaf area index (LAI) was highest in the treatment with 20 cm intrarow spacing and 60 cm inter-row spacing, and a population of 125,000 plants ha-1. Photosynthetically active radiation (PAR) levels were significantly higher in the treatment with 20 cm intra-row spacing and 60 cm inter-row spacing, attributed to early and complete ground cover. Yield attributes like cob weight was not influenced by the treatments. However, cob length, cob girth, number of grains, and test weight were maximized in the treatment with 40 cm intra-row spacing and 100 cm inter-row spacing, with the lowest population. The highest grain yields were achieved with 20 cm intra-row spacing and 60 cm inter-row spacing, and a population of 125,000 plants ha-1. Similar yields were obtained with 40 cm intra-row spacing and 60 cm inter-row spacing, with a population of 100,000 plants ha-1. Nutrient and protein content in the plant were not significantly affected by the treatments. Stover yield was also higher in the treatment with 20 cm intra-row spacing and 60 cm inter-row spacing. Nitrogen, phosphorus, and potassium uptake were significantly higher for 20 cm intra-row spacing and 60 cm inter-row spacing due to increased dry matter accumulation. From economic perspective, the treatment with 40 cm intra-row spacing and 60 cm inter-row spacing was the most cost-effective option, with lower cultivation costs and higher net income at both the locations (PAU, Ludhiana and RRS, Gurdaspur).ThesisItem Restricted Genome wide association studies for sorghum shoot fly and red leaf spot resistance(Punjab Agricultural University, 2024) Jasneet Kaur; Dr. R.S. SohuThe present study was conducted for the assessment of shoot fly and red leaf spot resistance and discovery of markers and candidate genes associated with target traits in Sorghum bicolor across years and locations. A diversity set of 121 germplasm lines, acquired from ICRISAT and IIMR, was evaluated under two environments (two locations – Ludhiana (2020-21 and 2021-22) and Faridkot (2021-22)) for shoot fly whereas at Ludhiana for two years kharif 2020-21 and 2021-22 for red leaf spot disease. The germplasm lines were evaluated for nine morphological traits for shoot fly representing- per cent dead heart formation (DH), glossiness(GL), leaf sheath pigmentation (LSP), plants infested with eggs (PWE), number of eggs per plant (PPE), seedling vigour (SV), seedling height (SDH), plant height (PH) and number of tillers (TL). Significant genetic and genotype x environment interaction variance was observed for all test traits. Ten genotypes viz., IC 288122, IS 7657, IS 7663, IS 7745, IS 8155, IS 8282, IS 8256, IS 8348, IS 8230 and SSV 17 were found resistant and gave stable performance across all environments. Resistant genotype IS 8348 was selected for generation of bi-parental cross with susceptible genotype IS 7017. Similarly, disease score and area under disease progressive curve was used for assessment of red leaf spot disease. Significant genetic variance and genotype x environment interaction variance was observed for disease score under both the environments. Four lines viz., IS 7017, IS 8015, IS 8256 and SCL 4 were free from infection, indicating resistance to red leaf spot. Best linear unbiased predictors values (BLUP) of traits were used for genome-wide association study (GWAS) using 42678 SNPs. Linkage disequilibrium decay in population was estimated to be ~50kb. A large number of significant marker-trait associations were identified for all test traits. The genomic regions identified on chromosomes 1, 2, 4, 7 and 10 were previously reported to be associated with shoot fly resistance for trait DH and GL. Annotations helped to predict candidate genes in the genomic regions associated with the significant SNPs. Important candidate genes involved were receptor kinases (LRR, serine/threonine protein kinases, hydrolases and transferases), stress signalling pathways (ethylene, abscisic acid and jasmonic) for shoot fly and (RLP, WRKY, F-box protein, CDP, zinc finger and putative disease resistance protein) for red leaf spot disease. Associative SNPs were converted into KASP markers and S1_15319241 KASP marker associated with DH was validated in the F2 population of cross IS 8348 x IS 7017.ThesisItem Restricted Identification of major abiotic bottlenecks in Wheat (Triticum aestivum L.) productivity in Punjab(Punjab Agricultural University, 2024) Mahajan, Sakshi; Dr. Prabhjyot Kaur SidhuThe field experiment entitled, “Identification of major abiotic bottlenecks in Wheat (Triticum aestivum L.) productivity in Punjab” was carried out at the Research Farm of the Dept of Climate Change and Agricultural Meteorology, PAU, Ludhiana during rabi 2022-23. Five wheat cultivars (Unnat PBW343, Unnat PBW550, PBW826, PBW869 and PBW824) were sown under six thermal environments (D1: 26th October, D2: 2nd November, D3: 9th November, D4: 16th November, D5: 23rd November and D6: 30th November) in a split plot design. The perusal of data revealed that the number of days required to complete the phenological stages were more under D1 followed by D2, D3, D4, D5 and D6. The cv Unnat PBW550 took least number of days to complete its life cycle. The LAI, tiller count periodic dry matter and PAR interception was more under thermal environment D1 and cv PBW826 while these parameters were more under late sown conditions in cv Unnat PBW550. The grain yield of wheat cultivars was reduced with delay in sowing by ~34 days (26th October to 30th November) by 40.7, 39.1, 46.0 and 42.6% in cv Unnat PBW343, PBW826, PBW869 and PBW824, respectively while in cv Unnat PBW550 the yield increased by 23.9% with delayed sowing. The GDD and PTU had positive relation with the yield and yield attributes amongst the four cultivars, while a negative relation was found with the cv Unnat PBW550. The weekly and monthly thumb rule models were developed using past 15 years meteorological data. It was concluded that the temperature and sunshine hours decreased from October till January and later from February onwards increased till April and the inverse happens with RH. The crop weather calendars that were developed will provide useful information regarding meteorological data for different growth stages and can be used to issue agro-advisories for crop yield prediction. The analysis revealed that high temperature was the cause of low yield during 2021-22 while rainfall during grain filling of wheat contributed to yield loss during 2014-15. Maximum/minimum temperature of 16-27/0-12oC, 14-23/3-10 oC and 25-36/10-20 oC during vegetative, flowering and physiological maturity stages, respectively were favourable for high yield of wheat in Punjab.
