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Chaudhary Charan Singh Haryana Agricultural University, Hisar
Chaudhary Charan Singh Haryana Agricultural University popularly known as HAU, is one of Asia's biggest agricultural universities, located at Hisar in the Indian state of Haryana. It is named after India's seventh Prime Minister, Chaudhary Charan Singh. It is a leader in agricultural research in India and contributed significantly to Green Revolution and White Revolution in India in the 1960s and 70s. It has a very large campus and has several research centres throughout the state. It won the Indian Council of Agricultural Research's Award for the Best Institute in 1997. HAU was initially a campus of Punjab Agricultural University, Ludhiana. After the formation of Haryana in 1966, it became an autonomous institution on February 2, 1970 through a Presidential Ordinance, later ratified as Haryana and Punjab Agricultural Universities Act, 1970, passed by the Lok Sabha on March 29, 1970. A. L. Fletcher, the first Vice-Chancellor of the university, was instrumental in its initial growth.
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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 Studies on the salt tolerance status of clusterbean [Cyamopsis tetragonoloba (L.) Taub.] and its improvement by in vitro methods by using ethylmethane sulphonate(CCSHAU, 2013) Deepika; Dhingra, H.R.Present investigation was conducted on clusterbean (Cyamopsis tetragonoloba (L).Taub.) var. HG 2-20 to study the effect of chloride dominated salinity on growth, development and yield attributes and to explore the possibility of induction of salinity tolerance through deployment of EMS. Seedling emergence, plant height, root length and its fresh as well as dry weights decreased with progressive increase in rhizospheric salinity. Membrane injury of leaves and roots, leaf succulence increased while relative water content and chlorophyll fluorescence decreased with salinity. Accumulation of soluble sugars and free proline in leaves with salinity is indicative of osmotic adjustment of leaves to some extent but decline in protein and starch content is possibly due to ion toxicity. Salinity decreased male fecundity conomittant with reduction in quantitative production of ovules. This is suggestive of female adjustment according to male fecundity and resource availability as evident from no change in the % ovules fertilized under saline conditions over control. Salinity decreased number of pods/cluster, number of pods/plant and number of seeds/pod. However percent pod set did not differ significantly upto 8dSm-1 and then decrease was evident at 12 dSm-1 level. Dry aerial biomass/plant was not affected significantly of 4dSm-1 salinity and it decreased with further increase in the level of rhizospheric salinity. Harvest index was not affected significantly by salinity upto 8dSm-1 level and then it declined at 12dSm-1 salinity. Seed yield/plant and seed quality in terms of starch, protein and guar gum content was, however deteriorated by rhizospheric salinity. Protein profile of leaves of guar plants raised under non-saline conditions showed polypeptide bands ranging from 67.61, 45.28, 29.12, 21.38 and 14.45 kDa. Among these, polypeptide bands of MW 45.28 and 14.45 kDa were present in leaves of salinized plants. Among various explants, cotyledonary node was found be the best explant and MSB5 medium + 2mgl-1 2,4-D and 1mgl-1 BAP supported best callus growth. Callus of cotyledonary node origin was compact, while those from other explants were fragile and forms suspension in EMS solution. BAP (1 and 2 mgl-1) alone supported 100% callusing from the cut end cotyledonary node while 2.0 mgl-1 BAP in combination with 1.5 mgl-1 NAA supported minimum callusing. Various permutation and combinations of plant growth regulators were used for regeneration from callus but could not be successful. Among different concentrations of salt, 200 mM NaCl was found to be the lethal. Culturing of EMS treated calli on NaCl adjunct medium showed no significant growth of callus upto 2.0h EMS treatment. An increase of duration of EMS treatment beyond 2.0h supported growth of callus which increased upto 3.0h and again declined with further increase in duration of EMS treatment. Fresh and dry weights of calli treated with EMS also increased upon raising on salt free medium over untreated ones. On the other hand, it decreased in untreated and treated calli (upto 2.0h duration) upon culturing on salt amended medium. EMS treatment of 2.5h and 3.0h duration improved callus growth on salt amended medium. EMS treatment in general increased starch, protein content, proline content and sodium content on salt amended medium over respective control while it decreased chloride content over respective control. Untreated subcultured callus, showed polypeptides of MW ranging 67.61, 66.07, 56.23, 50.12, 32.36 and 18.20 kDa, while the fresh callus showed 5 polypeptides of MW ranging 67.61, 50.12, 23.93, 21.38 and 13.80 kDa. NaCl induced new bands of MW 57.54, 28.18, 26.92 and 20.89 kDa while EMS treated calli for 2.5h and 3.0h and cultured on salt free medium showed polypeptide band of MW 56.23 and 32.36 kDa. Culturing of these calli on salt amended medium revealed presence of polypeptide bands of MW 67.61, 53.70, 50.12, 28.84, 20.89 and 18.62 kDa.ThesisItem Open Access Studies On Reproductive Characters Inerspecific Hybridization And Plant Regeneration In Cyamopsis Species(Chaudhary Charan Singh Haryana Agricultural University, College Of Basic Sciences And Humanities : Hisar, 2011) Ahlawat, Anju; Dhingra, H. R.ThesisItem Open Access Effect Of Saline Irrigation On Anatomical, Reproductive And Physiological Aspects Of Chickpea (Cicer Arietinum L.) Genotypes(Chaudhary Charan Singh Haryana Agricultural University; Hisar, 2010) Sunder Singh; Dhingra, H RThesisItem Open Access Effect of salt stress on the morphophysiological, yield and quality parameters in isabgol (Plantago ovata Forsk.) genotypes(CCSHAU, 2014) Suraj Kala; Varshney, U.K.Effect of NaCl salt stress on the morpho-physiological, yield and quality parameters in four isabgol (Plantago ovata Forsk.) genotypes viz. GI-2, HI-96, PB-80 and HI-5 was studied under screen house conditions. All the genotypes were grown in the dune sand filled polythene bags with control (without salt), 5 and 10 dSm-1 EC level of NaCl salt solution along with nutrients. Sampling was done at vegetative (58 DAS), flowering (115 DAS) and maturity (150 DAS) stage of growth. Results revealed a significant reduction in various plant growth parameters at every growth stages with increasing salt stress. Root/shoot ratio remained indifferent with the increasing salt stress level up to flowering stage but an increase in the ratio was observed at the maturity stage. Both reproductive effort as well as reproductive capacity of all the isabgol genotypes decreased with increasing salt stress. Reduction in plant growth, in general was maximum in the genotype PB-80 and minimum in GI-2. Increasing salt stress caused reduction in various yield attributes in all the isabgol genotypes. Mucilage (%) as well as seed swelling factor of all the isabgol genotypes also significantly declined under salt stress. Decline in yield as well as quality parameters was highest in the genotype PB-80 and lowest in GI-2. On the other hand increasing EC level caused enhancement of RMI, lipid peroxidation and ROS content in all the isabgol genotypes. The increase was maximum in the genotype PB-80 and minimum in GI-2. Chlotrophyll, carotenoid, ascorbic acid and total soluble protein content in leaves of all the isabgol genotypes decreased with increasing salt stress level and maximum decrease was found in the genotype PB-80 and minimum in GI-2. An accumulation of proline and total soluble carbohydrate content in leaves of all the isabgol genotypes was detected with the increase of salt stress; increase being maximum in the genotype GI-2 and minimum in PB-80. The enhancement in the activity of antioxidant enzymes viz., CAT, SOD and POD with the increase of salt stress was also highest in GI-2 and lowest in PB-80 among all the genotypes. In the Protein profile of leaves of the genotype GI-2 three new protein bands of the MW 29.59, 54.95 and 55.72 kDa and in the genotype HI-96 two new protein bands of the MW 29.59 and 55.72 kDa were detected under salt stress. While in the genotype PB-80 one protein band of MW 30.06 kDa was disappeared and one new protein band of MW 43.65 kDa was appeared under salt stress. Genotype HI-5 showed three new protein bands of MW 43.65, 55.72 and 66.07 kDa and disappearance of one protein band of MW 32.36 kDa under salt stress. The de novo synthesis of new protein bands under salt stress could be responsible for induction of salt tolerance in isabgol genotypes. While in the genotype PB-80 and HI-5 disappearance of protein bands may be viewed as marker of their salt sensitivity. Sodium and chloride content in different plant parts of isabgol genotypes increased while potassium content decreased with increasing EC level. Maximum sodium and chloride content was found in the genotype PB-80 and minimum in GI-2. Potassium content on the other hand was highest in the genotype GI-2 and lowest in PB-80. Based on the overall performance of all the genotypes under present study with respect to various morpho-physiological, yield and quality parameters, the genotype GI-2 was found to be most tolerant and the genotype PB-80 least tolerant to salt stress. Relative order of salt tolerance of different isabgol genotypes was GI-2>HI-96>HI-5>PB-80.