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ThesisItem Embargo Morpho-physiological, biochemical and molecular changes in BMR (brown mid-rib) sorghum under rainfed conditions(Punjab Agricultural University, 2024) Gurwinder Kaur; Dr. Harpreet Kaur OberoiIn the present investigation, two BMR (CSV 43 and SPV 2017) and non-BMR (CSV 15 and SPV 462) sorghum types were studied under irrigated and rainfed conditions at 45, 55, 65, and 75 DAS for morpho-physiological and biochemical parameters. Rainfed condition decreased green and dry fodder yields, along with morpho-physiological parameters in both sorghum types. Non-BMR genotypes exhibited higher yields under both irrigated and rainfed conditions. Higher SOD, POX, CAT and total antioxidant potential (FRAP, DPPH, TRP and HFRS) were observed under rainfed condition with lower H2O2 and MDA content in BMR genotypes over the non-BMR genotypes. Nitrogen assimilation enzymes (NR, NiR and GOGAT) decreased, while nitrate and nitrite N-content increased under rainfed condition in BMR and non-BMR sorghum. Rainfed condition increased the PAL, TAL activity and phenolic compounds in BMR genotypes compared to non-BMR genotypes. Moreover, due to better defence mechanism in BMR genotypes, fodder quality in terms of crude protein, crude fat, and IVDMD parameters was noticed to be higher as compared to non-BMR genotypes. Gene expression analysis revealed increased expression of stress-responsive genes (Gly-1, Gly-2, D-LDH, PAL, rubsico, SucS, stay green) in both genotypes under PEG 8000-induced drought stress. BMR genotype exhibited higher fold change in Gly-1, Gly-2 and SucS gene expressions, contributing to enhanced detoxification and photosynthetic stability. The present study suggests that BMR genotypes (CSV 43 and SPV 2017) compared to non-BMR sorghum genotype (CSV 15 and SPV 462) showed improved growth, enzymatic activities and gene expression related to defence to counteract the negative impact of rainfed conditionThesisItem Embargo Identification of low oxalate lines in cherry tomato using biochemical and molecular tools(Punjab Agricultural University, 2022) Simarjot Kaur; Chawla, NeenaThe objectives of the present study were to evaluate different biochemical components in sixty genotypes of cherry tomato namely, total soluble solids (TSS), lycopene, total carotenoids, vitamin C, titratable acidity, total antioxidant capacity, oxalate along with enzymes involved in oxalate biosynthetic pathways. Further, the expression analysis of oxalate biosynthetic pathway genes was carried out in selected genotypes of cherry tomato To study the biosynthesis of an anti-nutrient ‘oxalate’, leaf and root tissue of nine genotypes out of sixty genotypes were selected on the basis of low, moderate and high oxalate content to evaluate variation in the enzyme activity of two out of four enzymes mediating the biosynthesis of oxalate. The enzymes studied were: Ascorbate peroxidase (APX) and Glycolate oxidase (GLO) at three developmental stages of cherry tomato plant. The study showed that enzymes APX and GLO increased from stage 1 (plant vegetative stage) to stage 3 (fruit maturation stage) in leaf and root tissue of the majority of genotypes but in some genotypes, the enzyme activity was found to increase from stage 1 (plant vegetative stage) to stage 2 (fruit colour turning stage) and then showed a decline from stage 2 (fruit colour turning stage) to stage 3 (fruit maturation stage). Overall, the enzyme activity was found prominent in leaf tissue as compared to that in root tissue. Furthermore, three out of nine genotypes were selected on the basis of low, moderate and high activity of both the enzymes as well as oxalate content. These selected genotypes were analyzed for the expression study of one APX gene (SlAPX) and one GLO gene (SlGLO) out of cluster of genes encoding for the enzyme activity. The maximum relative fold expression of SlAPX gene was reported in the leaf tissue at stage 1 i.e., plant vegetative stage in two genotypes i.e., FB-3-5 and PTC-2 except genotype RSC-4 wherein higher fold expression was found in root tissue than the leaf tissue that was followed by undetectable fold expression of the gene at later stages of plant development. In the case of SlGLO gene, the maximum fold expression was observed in leaf tissue as compared to root tissue at stage 2 (fruit colour turning stage) with the exception of RSC-4 genotype with higher fold expression of SlGLO gene in root tissue than leaf tissue. The SlGLO gene was found as one of several genes known to encode for oxalate biosynthesis whereas SlAPX gene was found independent of the APX enzyme activity as well as oxalate accumulation.ThesisItem Restricted Effect of priming with Moringa oleifera leaf extract on sorghum under salt stress(Punjab Agricultural University, Ludhiana, 2022) Anand Kumar; Oberoi, Harpreet KaurIn the present study, seeds of fourteen sweet sorghum genotypes were treated with different NaCl concentrations (0, 50, 100, 150, 200 mM). All germination traits decreased with increased salinity. Based on germination traits and dendogram obtained at 100 mM NaCl, PHULE VASUNDHARA and SPV 2074 were selected as tolerant and susceptible genotypes, respectively. Seeds of selected genotypes were primed with 10% Moringa oleifera leaf extract and grown in Hoagland‟s solution with and without 100 mM NaCl concentration. NaClinduced the activities of SOD (in roots), POX and PPO and reduced the activities of NR, NiR and GOGAT, however, significant improvement in the activities were observed with MLE in shoots and roots of two contrasting genotypes at 10 DAG. Salt stress resulted in increased Na+/K+ ratio, H2O2, MDA, sugars, phenols and decreased total chlorophyll. Gene expression of antioxidative enzymes was upregulated under salt stress in shoots and roots of both primed genotypes. Salt stress resulted higher PPO, GOGAT, NiR and NR gene expressions in shoots of both contrasting genotypes with priming conditions. Expression of HAK gene was higher in primed SPV 2074 shoot and root without salt stress. In both tissues of tolerant and susceptible genotypes, expression of HKT-6 was higher with and without priming under both control and salt stress conditions. Therefore, the present investigation suggests that moringa seed priming impact the growth, enzymatic activities as well as gene expression under salt stress in sweet sorghum.ThesisItem Embargo Assessment of bio-ethanol production potential from juice using sugarcane varieties(Punjab Agricultural University, 2023) Gupta, Alisha; Kashyap, LenikaIn the present study, 22 different sugarcane varieties/clones belonging to different maturity groups (early and mid-late) were procured from the Regional Research Station, Kapurthala and screened for 8 cane yield and its component traits, 6 physiological traits and 8 juice quality traits. Six potential clones/varieties, namely, CoPb 95, CoPb 92, Co 0118, CoPb 93, CoJ 88, Co 0238 were identified for bioethanol production on the basis of leaf area (cm2), fresh weight (kg), brix (%), extraction (%) and fibre (%) cane and further evaluated for seven biochemical traits and bioethanol production potential under lab scale. Yeast strain (S. cerevisiae MK 680910) was used for bioethanol production under lab scale and scale-up studies. Varieties CoPb 92 (early maturing) and CoJ 88 (mid-late maturing) with higher initial sugars and ethanol content under lab scale were selected as potential sugarcane varieties for scale up studies. CoPb 92 variety with initial °brix (20.97°B), total sugars (20.05 g/100ml), reducing sugars (0.567 g/100ml) yielded 12.44 (% v/v) initial ethanol content with fermentation efficiency of 96.21 (%) and ethanol yield 0.50 (g/g). After first distillation of fermentation wort, 0.340 l ethanol was experimentally recovered from 1.04 l with 32.76 (% v/v) ethanol content. Second distillation recovered 0.329 l ethanol with final ethanol content of 78.46 (% v/v). Whereas, variety CoJ 88 with initial brix (20.24°B), total sugars (19.88 g/100ml) and reducing sugars (0.814 g/100ml juice) yielded initial ethanol content of 11.68 (% v/v) with fermentation efficiency of 94.49 (%) and ethanol yield of 0.47 (g/g). Recovery of 0.481 l ethanol was obtained after first distillation of fermentation wort and subsequent second distillation resulted in 0.301 l ethanol recovery with ethanol content of 75.45 (% v/v). This information generated can be added as basic input to scale up technical process towards enhancing the production of bio-fuel in sugar industries of the state associated in bioethanol production. So, future study can be planned to evaluate bioethanol production for extended period (October – March), when a mix of reducing sugars and non-reducing sugars concentrations are available as a fermenting substrate to be exploited by different fermenting bioagents.The sugar industies can thus work on dual mode: by switching over from sugar production to ethanol production; depending on the situation (sugar deficit/sugar surplus).ThesisItem Embargo Effect of popping and malting on nutritional characteristics in millets(Punjab Agricultural University, 2023) Harsimranjeet Kaur; Oberoi, Harpreet KaurThe present investigation aimed at evaluating the effect of popping and malting on nutritional characteristics in millets. Five genotypes each of sorghum, finger millet and pearl millet were analyzed after popping and malting process. The physical properties, nutritional characters, proximate analysis, bioactive components, enzymatic and antioxidant activities were observed in raw, popped and malted millet flours. The crude protein and energy were found to increase when popped and decrease after malting whereas crude fibre content significantly decreased in popped and malted flours of all millets over the raw flours. Non-significant reduction in fat content with significant hike in total soluble carbohydrates after processing of raw millets was seen. Malting resulted in increase of enzymatic activities (Lipoxygenase and alpha-amylase). Alkaloids and antioxidants (FRAP, DPPH and Ascorbic acid) increased whereas starch and amylose decreased after processing techniques compared to raw flour. Total phenols and tannins increased and reduction in antinutrients i.e. phytic acid, saponins and oxalate was seen in processed millet flours over raw. Millet genotypes showed differential protein banding pattern with lesser number of bands after processing of raw millets, however, 52 kDa band was observed in all processed millet flours. The results showed that the household processing techniques i.e. popping and malting improved the nutritional composition and antioxidant potential with simultaneously decrease in antinutritional components in all millets genotypes with pearl millet found to be slightly better and could fulfill the nutritional needs of poor community. Further, processed millet flours could be utilized in development of value added products.ThesisItem Restricted Biochemical basis of defense response in pigeonpea against maruca vitrata (fabricius) infestation(Punjab Agricultural University, 2022) Sukhmanpreet Kaur; Grewal, Satvir KaurThe present investigation was carried out to identify the biochemical basis of Maruca vitrata (pod borer) infestation in pigeonpea. Two pigeonpea genotypes, AL 1747 (moderately resistant) and MN 1 (susceptible) were compared for methylglyoxal detoxification, proline metabolizing enzymes along with free radical scavenging activities in leaves, flowers and pods under control and infested conditions. Lower accumulation of methylglyoxal (MG) in AL 1747 genotype was due to higher activities of enzymes of glyoxylase (Glyoxylase I, Glyoxylase II, Glyoxylase III) and non-glyoxylase (methylglyoxal reductase, MGR) pathway that converts MG into lactate. Higher activity of lactate dehydrogenase (LDH) in AL 1747 indicates the utilization of end product of MG detoxification pathway for energy production. Higher glutathione content in AL 1747 genotype might be responsible for efficient working of MG detoxification pathway under insect infestation. Higher activity of γGCS in AL 1747 genotype maintains the glutathione pool, necessary for the proper functioning of glyoxylase pathway to carry out the detoxification of methylglyoxal, thereby enhancing resistance. Higher activities of GST and GPX in AL 1747 genotype might be responsible for detoxification of toxic products that accumulates upon insect infestation. However, decreased activities of glyoxylase enzymes and MGR in MN 1 resulted in accumulation of MG which limits plant growth and development. The reciprocal regulation of proline dehydrogenase (ProDH) with proline biosynthesizing enzymes such as ornithine aminotransferase (OAT), glutamate dehydrogenase (GDH), Δ1-pyrroline-5-carboxylate synthetase (P5CS) in AL 1747 suggests that proline accumulation was initiated as part of induced defense response against M. vitrata infestation. Higher free radical scavenging activities such as 2,2-diphenyl-1-picryl hydrazyl (DPPH), superoxide anion, hydroxyl ion, nitric oxide, Ferric reducing antioxidant power (FRAP) might be responsible for lowering ROS accumulation upon insect infestation. Hence, the resistance of AL 1747 to M. vitrata might be due to higher activities of enzymes of glyoxylase and non-glyoxylase system; proline metabolizing enzymes as well as free radical scavenging activities thereby mitigating insect infestation induced oxidative stress.ThesisItem Restricted Exploring Garlic Biochemical Markers Against Root Knot Nematodes(Punjab Agricultural University, 2022) Shakya, Sandeep; Chawla, NeenaThe aim of the present study was to explore garlic biochemical markers and to evaluate garlic extract against root knot nematode of selected genotypes. The study included dry matter, total sugars, reducing sugars, starch, pyruvic acid, allicin and crude fibre in garlic of thirty genotypes. On the basis of allicin per cent, PG-114, PG-119 and PG-128 were selected as the best genotypes for evaluation of nematicidal properties of garlic extract keeping allicin as the main phytochemical. The garlic extract was made from three selected genotypes using different solvents (methanol and distilled water) of different concentrations (S1, S2, S3, S4 and S5). Out of these, PG-119 garlic extract prepared in methanol showed the best results in maximum egg hatching inhibition per cent over untreated control (84.01%, 83.63% and 82.57%) and juvenile mortality per cent (92.35%, 94.47% and 95.67%) after 24h, 48h and 96h respectively at S5 concentration (5.11 ug/ml) as compared to other concentrations applied for in-vitro observations. On the basis of the best performance of PG-119 under an in-vitro trial, it was selected for an in-vivo trial in which a single dose, as well as a double dose of S4 concentration (17.74 ug/ml), showed maximum reduction in root gall index (3.124 and 2.111), final soil population (381.791 and 284.323) and a number of egg mass of root-knot nematode (21.333 and 11.000) as compared to the untreated control. Also, maximum enhancement in plant growth such as plant height and plant weight were observed with the same concentration. Overall, the nematicidal properties of the garlic extract exhibited concentration and time-dependent behaviour.ThesisItem Embargo Biochemical and molecular shifts in Solanum melongena alien introgression lines following JASSID, Amrasca bigutulla bigutulla (Ishida) infestation(Punjab Agricultural University, 2023) Handa, Sysha; Rimaljeet KaurBrinjal jassid, Amrasca biguttula biguttula (Ishida) is emerging as devastating pest which significantly affects its yield and production. Varietal resistance could be an imperative and effective strategy to combat pest feeding. Vegetable specialist at PAU, Ludhiana developed S. melongena – S. insanum derived lines which displayed excellent variability to resistance against jassid infestation. Key resistance determinants at morphological, biochemical and transcriptional levels have not been explored so far. Therefore, we planned our research with the objective of determining jassid responsive defensive traits which encompasses phenotypic assessment, morphological characterization, biochemical alterations and gene expression profiling in S. melongena – S. insanum against jassid feeding. Phenotypic assessment of introgression lines (ILs) on the basis of jassid population per plant and pest percent intensity categorized genotypes into highly resistant, moderately resistant, moderately susceptible and highly susceptible genotypes. An increase in affected leaf area was determined with increase in the incidence of A. bigutulla bigutulla infestation. Genotypes with high trichome density and a short trichome length were resistant to pest attack. The increased expression of APX transcripts and ascorbate peroxidise (APX) activity along with a decrease in jassid count in resistant genotypes determined the pivotal role of APX in S. melongena genotypes against jassid attack. This candidate would be used in deployment of S. insanum resistance during brinjal breeding programs. On the contrary, susceptible genotypes showed higher expression of catalase (CAT) activity and CAT transcripts during jassid feeding. APX primarily decomposed H2O2 in resistant lines, whereas CAT primarily did so in susceptible genotypes. During high jassid infestation, H2O2 concentration rapidly increased in test genotypes. In comparison to resistant genotypes, H2O2 buildup was higher in susceptible genotypes. Increased total sugar and sucrose content in susceptible genotypes following jassid infestation suggests their contribution to the pest susceptibility. The genotypes with high jassid infestation displayed high oxidative stress (H2O2 and MDA), indicating that an increase in liposygenase (LOX) activity might cause high production of hydroperoxides, which in turn led to an increase in CAT, polyphenol oxidase (PPO), and phenolic accumulation. The association between JPP and H2O2, LOX, CAT, PPO, MDA, and total phenols was significant and positive. Thus, our findings suggest that susceptible genotypes experienced more oxidative stress than resistant genotypes.ThesisItem Restricted Development of near-infrared reflectance spectroscopy calibration equations for phytochemical profiling of Brassica juncea(Punjab Agricultural University, 2022) Mundi, Harshdeep Kaur; Sharma, SanjulaNear-infrared reflectance spectroscopy (NIRS) with chemometrics has emerged as an effective technique for simple, rapid, inexpensive, safe, authentic and non-destructive determination of different biochemical compounds in various food and agricultural products. It quantifies organic matter in the sample by recording the absorbance signals in the near-infrared light for the major chemical groups in less than a minute. Considering the advantages of NIRS, the present study was conducted to develop NIRS-based prediction model as a speedy alternative for measuring quality traits, otherwise assessed using chemical methods which are often timeconsuming labor-intensive and with dangerous effects on health. Brassica crop is well known for its health promoting bioactive compounds. The precise and rapid assessment of these bioactive compounds found in Brassica juncea L. (Indian mustard) may help in determining the genetic variation for these traits across diverse oilseed Brassica germplasm. This study utilizes seeds from 178 diverse B. juncea accessions harvested in the years, 2019-20 and 2020-21 which were estimated for oil content, fatty acid profile, tocopherols and phytosterols using wet chemistry analytical methods. Simultaneously, the spectra of the same samples were acquired as the logarithm of reciprocal of reflectance (log 1/R) in the entire UV-NIR wavelength range of 400 to 2498 nm of electromagnetic spectrum by scanning on NIR System model 6500, Inc., Laurel, MD, USA. Modified partial least-square (MPLS) regression method based NIRS models were developed, wherein the model exhibited higher coefficient of determination (RSQ) and coefficient determination of cross validation (1-VR) value ranging from 0.650 to 0.987 and 0.612 to 0.981 for all the traits except stearic acid, linolenic acid and phytosterol. The traits with high RSQ also possess lowest values of standard error of calibration (SEC), and standard error of cross-validation (SECV) ranging from 1.242 to 18.428 and 1.427 to 22.033. External validation of the developed models yielded high RSQ value for oil (0.693), eicosenoic acid (0.716), erucic acid (0.870), αtocopherol (0.711), γ-tocopherol (0.829) and total tocopherol (0.725) with standard error of prediction (SEP) ranging from 1.496 to 6.166. The statistical results obtained demonstrated the efficacy of newly developed NIRS model for accurately predicting the oil, fatty acids (eicosenoic acid, erucic acid) tocopherols (α-, γ-, total tocopherol) contents. The outcomes of the study will assist in precise screening and selection of superior quality lines in Brassica breeding programs.
