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ThesisItem Open Access Efficacy of gibberellic acid and triacontanol on morpho-physiological, biochemical and anatomy of Sorghum bicolor L. under salt stress(CCSHAU, Hisar, 2023-09) Mehra, Indu; Tallapragada, SrideviThe present study entitled “Efficacy of gibberellic acid and triacontanol on morpho-physiological, biochemical and anatomy of Sorghum bicolor L. under salt stress,” was examined in the screen house during the kharif season of 2022. Prior to sowing, different salt stress levels (control, 4, 6, and 8 dS m-1) were maintained by saturating each pot. Sorghum bicolor L. seeds were planted in pots under screen house conditions on July 25th, 2022. Triacontanol was applied at concentrations of 10 ppm and 20 ppm, while Gibberellic acid was applied (200 ppm and 400 ppm) to both stressed and non-stressed plants of two genotypes (CSH20MF and CSH24MF). Sampling was conducted at 30 and 60 days after sowing (DAS). Growth parameters were examined. An inverse relationship was observed between these parameters and increasing salinity levels. Furthermore, anatomical leaf characteristics were improved following the application of Triacontanol and Gibberellic acid, counteracting the negative effects of salt stress. A similar pattern was observed in the relative water content under different salt levels. Total chlorophyll content, carotenoid content, gaseous exchange studies, chlorophyll index, and photochemical quantum yield also displayed a decreasing trend from the control to the highest salt level (8 dS m-1) in both genotypes. The application of Triacontanol and Gibberellic acid aids in maintaining the relative water content, increasing proline and glycine betaine content, enhancing gas exchange, ash content, chlorophyll index, total chlorophyll, carotenoid content, anthocyanin, flavonoid content, and promoting the activity of antioxidant enzymes regardless of salt stress in both genotypes. Conversely, the relative stress injury worsens with each increment of salt stress, but the application of Triacontanol and Gibberellic acid partially mitigates the adverse effects of salt stress. An increase in Na+ content and a decrease in K+ and Ca2+ content was observed under salt stress, whereas the opposite trend was observed after the application of TRIA and GA3. In sorghum genotypes CSH24MF and CSH20MF performed better under salt stress and exhibited a superior response in alleviating salt stress with the foliar application of 20 ppm of Triacontanol.ThesisItem Open Access Efficacy of jasmonic acid on morpho-physiological, biochemical and quality attributes of sorghum (Sorghum bicolor L.) under salt stress(CCS HAU, Hisar, 2022-07-16) Prakriti; Sridevi TallapragadaThe present study entitled “Efficacy of jasmonic acid on morpho-physiological, biochemical and quality attributes of sorghum (Sorghum bicolor L.) under salt stress” was investigated in the screen house during the kharif season of 2021. Before sowing, the desired levels of salt stress (control, 5, 7.5 and 8 dS m-1) were maintained by saturating each pot. Seeds of Sorghum bicolor L. were grown in pots under screen house conditions on 16th July 2021. Jasmonic acid (4.5, 6.5 and 8.5 mM) was applied exogenously after 30 days after sowing (DAS) under stressed as well as non-stressed plants in both genotypes (HJ 541 and CSV 21F). Sampling was done at 40 and 60 DAS. Growth parameters decreased with increasing levels of salt stress, whereas foliar application of jasmonic acid enhanced the growth of plants. Similarly, anatomical parameters of leaf were enhanced after the application of jasmonic acid which were negatively affected by salt stress. Plant water relation, gaseous exchange studies, chlorophyll index and photochemical quantum yield also showed declining trend from control to 8 dS m-1 of salt level in both genotypes, but more percent decrease was observed at 8 dS m-1 of salt stress like in ψw (46.66%), ψs (18.27), photosynthetic rate (31.80%), transpiration rate (14.60%) and stomatal conductance (44.41%) in HJ 541, over their respective control. The values ranged from 7.32 to 6.66, 27.37 to 23.32 and 54.51 to 50.83 in protein, fibre and in-vitro dry matter digestibility, respectively, at 8 dS m-1 of salt stress in CSV 21F at 40 DAS. Application of JA help in the maintenance of plant water status, enhanced the exchange of gases and chlorophyll index irrespective of salt stress in both genotypes. Reversibily, electrolyte leakage and ash content enhanced with every increment of salt stress but application of JA ameliorate the adverse effect of salt stress to some extent. Increment in Na+ but a decrease in K+ and Ca2+ content was noted under salt stress. While, the reverse trend was noticed after application of JA. In nut shell, sorghum genotypes CSV 21F and HJ 541 performed better under salt stress and showed better response in alleviating salt stress with foliar application of 8.5 mM jasmonic acid.ThesisItem Open Access Response of jasmonic acid for alleviating cadmium (Cd) toxicity in Brassica spp. (Brassica napus, Brassica juncea)(CCS HAU, Hisar, 2022-07-22) Sajid Khan; Sridevi TallapragadaThe present study entitled “Response of jasmonic acid for alleviating cadmium (Cd) toxicity in Brassica spp. (Brassica napus, Brassica juncea)”. was investigated in the screen house during the Rabi season of 2021. Before sowing, the desired levels of cadmium stress (Control, 14 mg/kg, 21 mg/kg) were maintained by saturating each pot. Jasmonic acid (5 μmol L-1, 10 μmol L-1, and 20 μmol L-1) was applied exogenously after 30 days after sowing (DAS) under stressed as well as non-stressed plants in both genotypes (RH 725 and HNS 0901). Sampling was done at 45 DAS. Seeds of Brassica spp. were grown in pots (three plants per pot) under screen house conditions on 15th October 2021. Growth parameters decreased with increasing levels of cadmium stress, whereas foliar application of jasmonic acid enhanced the growth of plants at sampling stages. Similarly, plant water relation, gaseous exchange studies, chlorophyll content (SPAD units) and oil content also showed declining trend from Control to 21 mg/kg of cadmium level in both genotypes. Per cent decrease was observed at 21 mg/kg of cadmium stress like in ψw (68.3%), ψs (52.2%), photosynthetic rate (30%), transpiration rate (25%) and stomatal conductance (29.2%) in RH 725 at 45 DAS, over their respective control. The values ranged from 31.66 to 24.47 and 81.27% to 60.95% of chlorophyll content (SPAD units) and RWC respectively, decreased at 21 mg/kg of cadmium stress in RH 725 at 45 DAS. In anatomical study it is observed that increase in cadmium level is responsible for decrease in stomatal frequency (23.9%, 30%) and stomatal index (25.9 9%, 29.01%) in RH 725 and HNS 0901 respectively at 21 mg/kg of cadmium stress. Foliar application of jasmonic acid is responsible for slightly increase in stomatal frequency and stomatal index at its maximum value (20 μmol L-1) in both genotypes at 45 DAS. Application of JA help in the alleviation of plant water status, exchange of gases and chlorophyll pigment irrespective of cadmium stress in both genotypes at 45 DAS. Reversibly, ash content and ECe show increase along different cadmium level. Seed yield decline significantly by decreasing yield attributing characters i.e., siliqua per plant, seed per siliqua, biological yield under cadmium stress. Reduction in seed yield was ranged from 4.55 to 3.14 in RH 725 and 4.16 to 2.58 in HNS 0901 at 21 mg/kg. Uptake of cadmium from soil increase along with increase in cadmium level 14 mg/kg to 21 mg/kg is 1144.50 μg/pot to 1450 μg/pot in RH 725 and 1025 μg/pot to 1224.8 μg/pot in HNS 0901. JA is showing an alleviating effect against cadmium uptake from soil and decrease cadmium uptake in plant. Conclusively, based on the above studies, it can be concluded that after foliar spray of JA, RH 725 performed better under cadmium stress by maintaining higher plant growth, water status, photosynthetic rate, seed yield, biological yield. JA treatments not only mitigate the negative effect of cadmium stress on plants, but also showed, a stimulating effect and 20 μmol L-1 JA was found more effective than 10 μmol L-1 at 45 DAS.ThesisItem Open Access Wide hybridization and embryo rescue for pre breeding in Indian Mustard(CCSHAU, Hisar, 2022-10) Anjali; Lakra, NitaThe present study was conducted to develop hybrid between Brassica juncea cv. RH 725 taken as a female parent and Sinapis alba genotype DRMR 2183 taken as a male parent with the help of embryo rescue because hybrid embryo usually abort at early stages. The pod setting was 40%. The hybrid siliquae were intermediate in size with respect to the parent plant with an average length of 3.44 cm. The average length of siliquae in B. juncea cv. RH 725 was 4.85 cm and in Sinapis alba the average length was 1.92 cm. Developing ovules were used as an explant and cultured on Murashige and Skoog‟s medium supplied with optimum concentration of different growth regulators. Maximum no. of ovules germinated after 25-30 days of inoculation. Maximum shooting response (48.33%) was observed on MS medium supplemented with MS + BAP (2.5mg/l) + CH (0.5g/l) and maximum rooting response was observed on MS + IAA (0.5 mg/l) i.e. 57.17%. After hardening in a mixture of sterilized sand: soil (1:1), 22% plants successfully moved to green house.ThesisItem Open Access Study on sustainability and survival rate of in–vitro propagated sugarcane under green house conditions(CCSHAU, Hisar, 2021-09) Shweta; Tallapragada, SrideviThe investigation was carried out to study the effect of different energy sources and gelling agents on in vitro multiplication and rooting response on sugarcane cultivar Co-89003. For in vitro multiplication plant cultures were inoculated on MS media supplemented with various sugars (fructose, sucrose, glucose, table sugar and maltose) and gelling agents (agar-agar, agarose, gelrite, guargum and isabgol). The maximum shoots (11.5±0.23 and 9.7±0.23) were observed on medium enriched with table sugar and agrose, respectively. And the maximum shoot-length (7.1±0.39 cm and 7.1±0.51cm) was noticed in sucrose and agar –agar media on after 28th day of inoculation. The very good growth/ rooting (+++++) was observed on ½ MS media supplemented with sucrose and gelrite. Then well rooting plantlets were transplanted into pots containing various compositions of soil, sand, vermicompost, FYM, coco pit and perlite for acclimatization. The maximum and survival (100%) were observed in four different potting mix including sand : soil : FYM, Sand : Soil : Vermicompost, Sand : Soil : FYM : Vermicompost and Coco pit : Vermicompost : Perlite under green house conditions. Hardened plantlets were successfully transferred to the field conditionsThesisItem Open Access Evaluation of antagonistic effect of cell free culture filtrates of Rhizoctonia sp., anastomosis groups/sub-groups infecting Zea mays(CCSHAU, Hisar, 2021-08) Sumit; Vimla SinghThe present study entitled “Evaluation of antagonistic effect of cell free culture filtrates of Rhizoctonia sp., anastomosis groups/sub-groups infecting Zea mays” was conducted during Kharif season of 2020 at CCSHAU, Regional Research Station Uchani, Karnal. Maize (Zea mays L.) is the most economically important cereal crops with a promising yield potential and industrial importance. Banded leaf and sheath blight (BLSB) disease caused by Rhizoctonia solani f.sp. sasakii is a devastating stress on Maize and which leads to huge monetary losses to the farmers. Single hypha tip culture of the isolates viz., Rs-Khp (Kohlapur), Rs-Rch (Ranchi), Rs-Bhw, (Bhuvneshwar), Rs-Kau (Kaul) and Rs-Lck (Lucknow) maintained on PDA plates and characterized for virulence profiles based on cultural, morpho metric and pathogenic variability. The R. solani isolates Rs-Khp, Rs-Kau and Rs- Lck were found to be virulent and comparatively more aggressive than isolates Rs-Rch and Rs-Bhw. Hyphal anastomosis studies showed that, weaker interaction of less virulent isolates with the more virulent isolates. Investigations on cross protective effects of cell free culture filtrates of weaker isolates in suppressing the virulent isolates was examined at dilutions 15% and 25%. Seeds pre-soaked in 15% dilution of culture filtrate showed better results in terms of reduction of severity of symptoms by 25%. However, satisfactory results with 25% dilution were not obtained. The findings reveal that culture filtrate of weaker isolated can serve to reduce yield losses due to severe isolates and such studies may be further undertaken at field level. Further, insight of mechanism of disease suppression may be studied to devise more accurate technique for minimizing losses due to BLSB.ThesisItem Open Access Morphogenetic and biochemical studies in callus cultures of devil’s thorn (Tribulus terrestris Linn.)(CCSHAU, 2013) Manogya; Goyal, S.C.Present investigation was carried out on Tribulus terrestris Linn. for the selection of suitable nutritive medium and explant for callus induction, growth and differentiation. In vitro studies were conducted with employing different explants viz. leaf, internode and node used for callogenesis and growth, node explant was found to be the best for callus induction and growth. Best callus growth was obtained on MS medium fortified with 2,4-D (2.0 mg/l) in combination with Kn (0.5 mg/l) depending on fresh and dry weight of callus. This callus selected for further studies of biochemicals and protein profile during differentiation. MS medium supplemented with BAP (3.0 mg/l) + NAA (1.5 mg/l) enabled the early regeneration of shoot from callus. Early root emergence from callus obtained on medium MS+NAA (2.0 mg/l). Biochemicals like starch and total soluble sugars decreased while reducing sugar, total phenol and total protein increased during root differentiation from callus. Enzyme activity viz. α- amylase, acid invertase, peroxidase and acid phosphatase increased during root differentiation. In shoot differentiating calli, all these metabolites were increased and enzyme activity of α-amylase and acid phosphatase increased while acid invertase and peroxidase activity decreased during shoot differentiation. SDS-PAGE studies revealed that there was synthesis of three root specific polypeptide bands with (MW 40.64, 19.23 and 11.81 kDa) and three shoot specific bands (MW 28.10, 17.02 and 12.46 kDa) during differentiation. MW 24.96 kDa and 21.20 kDa bands were common bands which disappeared during root and shoot differentiation. TLC analysis showed that the diosgenin and hecogenin were present in the calli as well as in natural fruits. They were further confirmed by IR spectroscopy. The incorporation of cholesterol in the medium was observed to be effective for the increased steroidal sapogenins (hecogenin and diosgenin) production in calli than natural fruits.ThesisItem Open Access Identification of physiological and molecular markers for salinity tolerance in wheat(CCSHAU, 2012) ASHA; Dhingra, H.R.The present investigation was carried out on eight varieties of wheat viz. WH157, WH1021, RAJ3765, KRL19, WH711, PBW550, WH542 and PBW502, to screen them for salinity tolerance on the basis of sexual characters and yield, to find biochemical, physiological and molecular basis of salinity tolerance under in vitro and in vivo conditions and to select and evaluate salinity tolerant somaclones. In the first experiment, all the eight varieties of wheat were subjected to a range of chloride dominated salinity viz. control 4, 8 and 12 dSm -1 to study its effect on physiological parameters like seedling emergence, chlorophyll fluorescence, membrane injury, photosynthetic rate, transpiration rate, water use efficiency, proline content and osmotic potential. In addition to these reproductive parameters like spike length, number of pollen/anther, pollen viability and in vitro pollen germination and yield and grain quality parameters like grain weight per spike, 100 grain weight, grain yield/plant, sedimentation value, hectoliter weight, starch and protein content of grain and mineral content (Na + , K + , Cl and SO 4 2-) were also recorded. Salinity decreased and delayed seedling emergence in all varieties. It also exercised inhibitory effect on plant height particularly in PBW550, WH542 and WH711. Salinity increased electrolyte leakage of flag leaf,increase being maximum in variety WH711 followed by PBW550 while minimum in WH1021 followed by KRL19. Photosynthetic rate, stomatal conductance and transpiration rate decreased with increasing saline irrigation in all varieties, but PBW550, WH542 and PBW502 suffered most. Water use efficiency (WUE) remained nearly unchanged under saline conditions. Salinity induced accumulation of proline in flag leaf of all tested varieties;percent accumulation being highest in WH157 followed by KRL19 and least in PBW550 and WH711 at highest level of salinity. The Ψs of flag leaf became more negative with increasing level of salinity stress. Number of spikes/plant, spike length, number of florets/spikelet decreased with salinity. Pollen production, pollen viability (%), in vitro pollen germination and tube growth were also adversely affected with increasing salinity; the effect being most pronounced in varieties PBW550and WH711. The yield parameters like number of grains/spike, grain weight/plant, 100 grain weight decreased with increasing salinity, however varieties KRL19 and WH1021 were less affected than others. Grain quality parameters like sedimentation xvi value and hectoliter weight decreased at higher salinity levels. Reduction for sedimentation value was more in PBW550 over other varieties and least in WH1021. Biochemical studies also revealed a decline in protein and starch content of grain with increasingsalinity. Analysis of mineral composition of seed showed a decrease in K + /Na + ratio and an increase in chloride and sulphate content. Minimum reduction for K + /Na + ratio at 12 dSm -1 salinity was observed in KRL19 followed by WH157 and RAJ3765. MS medium supplemented with 4 mg/l concentration of 2,4-D (MS1) was the best callusing and growth supporting medium for mature embryo. Culturing of actively growing callus on MS1 medium supplemented with a range of salinity (Control 4, 8 and 12 dSm -1 ) resulted in decline in RGR after 3 and 6 weeks of incubation. Varieties WH1021, WH157 and KRL19 possessed higher RGR than other varieties. Tissue culture studies adduced support to pot culture experiment for the parameters like osmotic potential, proline content, starch and protein content, K + /Na + ratio, Cl - and SO4 2- content at both stages of sampling i.e. after 3 and 6 weeks of incubation. Repeated sub-culturing of callus for 6 generations from sensitive varieties WH711 and PBW550 on MS1 medium with or without salinity, generated four different callus lines viz. control, salinity unadapted, salinity adapted and salinity adapted transferred to salinity free medium. RGR, osmotic potential, starch and protein content, K + /Na + ratio decreased while proline, Cl and SO 4 2-content increased in all salinity exposed callus lines as compared to control. Salinityadapted callus lines performed better than salinity unadapted callus lines for all parameters. Adapted calli when transferred to salinity free medium showed improved performance in respect of different evaluation parameters. Salinity induced synthesis of new polypeptide bands with M.W. 69.1, 36.3, 21.8, 17.3 and 15.1 kDa in salt tolerant WH1021 and in salt sensitive WH711,polypeptide bands with M.W. 79.4 & 75.8 kDa disappeared concomitant with synthesis of polypeptide bandswith M.W. 60.2, 36.3, 17.3 and 15.1 kDa.ThesisItem Open Access Physiological And Molecular Characterization Of Some Salinity Phytoremediating Plants(Chaudhary Charan Singh Haryana Agricultural University; Hisar, 2006) Sushma Rani; Goyal, S. C.
