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Dr. Rajendra Prasad Central Agricultural University, Pusa
In the imperial Gazetteer of India 1878, Pusa was recorded as a government estate of about 1350 acres in Darbhanba. It was acquired by East India Company for running a stud farm to supply better breed of horses mainly for the army. Frequent incidence of glanders disease (swelling of glands), mostly affecting the valuable imported bloodstock made the civil veterinary department to shift the entire stock out of Pusa. A British tobacco concern Beg Sutherland & co. got the estate on lease but it also left in 1897 abandoning the government estate of Pusa.
Lord Mayo, The Viceroy and Governor General, had been repeatedly trying to get through his proposal for setting up a directorate general of Agriculture that would take care of the soil and its productivity, formulate newer techniques of cultivation, improve the quality of seeds and livestock and also arrange for imparting agricultural education. The government of India had invited a British expert. Dr. J. A. Voelcker who had submitted as report on the development of Indian agriculture. As a follow-up action, three experts in different fields were appointed for the first time during 1885 to 1895 namely, agricultural chemist (Dr. J. W. Leafer), cryptogamic botanist (Dr. R. A. Butler) and entomologist (Dr. H. Maxwell Lefroy) with headquarters at Dehradun (U.P.) in the forest Research Institute complex.
Surprisingly, until now Pusa, which was destined to become the centre of agricultural revolution in the country, was lying as before an abandoned government estate. In 1898. Lord Curzon took over as the viceroy. A widely traveled person and an administrator, he salvaged out the earlier proposal and got London’s approval for the appointment of the inspector General of Agriculture to which the first incumbent Mr. J. Mollison (Dy. Director of Agriculture, Bombay) joined in 1901 with headquarters at Nagpur
The then government of Bengal had mooted in 1902 a proposal to the centre for setting up a model cattle farm for improving the dilapidated condition of the livestock at Pusa estate where plenty of land, water and feed would be available, and with Mr. Mollison’s support this was accepted in principle. Around Pusa, there were many British planters and also an indigo research centre Dalsing Sarai (near Pusa). Mr. Mollison’s visits to this mini British kingdom and his strong recommendations. In favour of Pusa as the most ideal place for the Bengal government project obviously caught the attention for the viceroy.
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ThesisItem Open Access Morphological and bio chemical characterization of Winged bean Rhizobium and its impact as seed Inoculant either singly or Combinedly with PGPR towards Yield Attributing Characters and Yield of Winged Bean (Psophocarpus tetragonolobus)”(Dr. Rajendra Prasad Central Agricultural University, Pusa, Samastipur, 2018) Kumari, Shweta; Yadav, R.C.A laboratory investigation for morphological and bio-chemical characterization and physiological properties of winged bean Rhizobium with a purified culture; and a field experiment consisting of four seed-inoculation treatments [No inoculation, Rhizobium, PGPR(Bacillus) and Rhizobium+PGPR(Bacillus)] and three nitrogen levels [0, 20 and 40 kg ha-1] in RBD (factorial) with three replications for evaluation of two winged bean germplasm IC-17002 and DLN-9, during crop season in the year 2014-15 and with better-performing IC-17002 in 2015-16 to find out impact of the Rhizobium as seed inoculant alone or in combinedly with PGPR on yield attributes and yield of winged bean were conducted at Tirhut College of Agriculture, Dholi Farm of RPCAU, Pusa (Bihar). The pure-culture of winged bean Rhizobium isolate was Gram-negative rod-shaped with a dimension of 0.59 μm x 1.81μm. The Rhizobium tested vis-à-vis (pea, gram, lentil and moong-rhizobia) showed a wide diversity in tolerance for its growth at different pH values, though neutral pH being the optimum. The winged bean-Rhizobium isolate produced 64.6 μg mL-1 of IAA at 72h of incubation with antibiotic-resistance and carbon-utilization test, the Rhizobium isolate was found to be resistant to vancomycin (30μg) and utilized a wider group of carbon sources than other rhizobia. The Fatty-Acid-Methyl-Ester (FAME) profiling confirmed that it belonged to Rhizobium-miscellany group. Seed inoculation and nitrogen levels greatly influenced nodule-parameters, nitrogenase activity and yield attributes, value of which were the highest with the combined inoculation of Rhizobium+PGPR (Bacillus), followed by seed inoculation of Rhizobium alone, and differed significantly among themselves at both 60 and 120 days after sowing (DAS). Nitrogen application @ 20 kg ha-1 recorded significantly higher nodule parameters over the control. The inoculation x nitrogen interaction effect was found to be significant with respect to nodule parameters. The combined inoculation of Rhizobium+PGPR (Bacillus) at 20 kg N ha-1 appeared significantly superior to other treatment combinations. S, Fe and Mo in the nodule recorded higher at 60 DAS than at 120 DAS. Rhizobium inoculation alone caused higher S and Mo content in nodule, whereas combined application of Rhizobium and PGPR recorded the highest Fe content at both growth stages. Nitrogen application @ 20 kg ha-1 increased Fe content significantly over no-nitrogen application at 120 DAS. The highest nitrogen content in pod was noted by combined inoculation of Rhizobium and PGPR treatment, followed by Rhizobium alone. Nitrogen @ 20 kg ha-1 was found to be superior to the higher level of N application. The combined inoculation of Rhizobium and PGPR at 20 kg N ha-1 proved better than the other treatments. The treatments recorded greater N and P content of winged bean plant at 60, 120 DAS and at harvest. In seed, Rhizobium and PGPR inoculation in combination recorded more N content than other treatments and was found to be at par with Rhizobium inoculation alone. Nitrogen application @ 20 kg ha-1 recorded higher N, P and K in seed of winged bean in first year only. Protein content in green edible pod, seed and tuber was significantly increased by combined inoculation of Rhizobium and PGPR over the control and was at par with the treatment of Rhizobium inoculation alone, whereas basal nitrogen @ 20 kg ha-1 favoured more protein content than higher level of nitrogen. After harvest of winged bean, the highest available soilnitrogen build-up was recorded with the treatment of Rhizobium inoculation alone. Seed-oil content (33.76 %) recorded the highest in the treatment with seed inoculation of PGPR at zero nitrogen application. The study proved the multiple-utilitarian potential of winged bean including its role in soil fertility improvement which was best with seedinoculation of Rhizobium-miscellany and PGPR either alone or in combination with basal dose of 20 kg N ha-1.ThesisItem Open Access Physiology of wheat (Triticum aestivum L.) under salt and high temperature stress condition(Dr. Rajendra Prasad Central Agricultural University, Pusa, Samastipur (Bihar), 2018) Neelambari; Singh, Ajay KumarUnder natural conditions, plants are often subjected to a combination of different stresses such as salt stress and heat shock. Wheat crops are slightly salinity tolerant but as a cool season crop it is sensitive to heat stress at both vegetative and reproductive stages. Recent studies revealed that the response of plants to a combination of two different stresses is specific and cannot be deduced from the stresses applied individually because increased transpiration rate associated with high temperature stress worsen the harmful effects of salinity stress. Therefore, the present investigation entitled “Physiology of wheat (Triticum aestivum L.) under salt and high temperature stress condition” was conducted with three main objectives (1) Screening of wheat genotypes for combined salt and high temperature stress to identify the contrasting sets of wheat genotypes on the basis of physiological traits. (2) To study physiological and biochemical mechanisms of tolerance of wheat genotypes subjected to independent, and combined salt and high temperature stress conditions. (3) To quantify the changes in yield and yield attributes of wheat genotypes grown under independent, and combined salt and high temperature stress conditions. In the present study, two independent experiments were conducted. The first experiment was done in laboratory condition where 46 wheat genotypes were screened for individual and combined salinity and high temperature stress condition at seedling stage. Wheat seeds were sown in petri plate in three replications for germination. A saline solution of composition NaCl:CaCl2: Na2SO4 (7:2:1) with salinity levels at 4.0 and 8.0 dS m-1 were used as irrigation solution for giving salt stress treatments. High temperature treatment, was given by keeping the petri dishes in incubator at temperature 37 ± 2°C, and combined salinity and high temperature stress was given by transferring salinity stressed plant to incubator at high temperature (37 ± 2°C). The physiological parameters were measured in 10-day-old seedling. Physiological parameters (germination percentage, shoot length, seedling length, shoot fresh weight, root fresh weight, seedling fresh weight, root dry weight, shoot dry weight, seedling dry weight, vigour index I, vigour Index II and SPAD value) were measured and found to be reduced, except root length which was found to be increased in some genotypes and decreased in others under salinity, high temperature and combined stress. However the reduction was more pronounced under combined stress. On the basis of physiological data, the contrasting set of wheat genotypes (tolerant genotypes, i.e., KRL-1-4, KRL-19 and HD-2733, and susceptible genotypes, i.e., HT-8, HI-1563 and HD-2987) were identified for further study. The genotypes identified from screening experiment were used for a pot culture experiment to study the physiological and biochemical mechanism of stress tolerance and yield/yield associated parameters. Three replications of pot were arranged in factorial completely randomized design. Four treatments were given, i.e., control (wheat genotypes were sown timely in soil with ECe 1.3 dS m-1), salinity stress treatment (wheat genotypes were sown timely in natural saline soil with ECe 7.4 dS m-1), high temperature stress treatment (wheat genotypes were sown late in soil with ECe 1.3 dS m-1) and combined salinity and high temperature stress treatment (wheat genotypes were sown late in natural saline soil with ECe 7.4 dS m-1). The physiological and biochemical parameters were measured in flag leaf at anthesis period and yield parameters were measured at physiological maturity. The result revealed a decrease in physiological parameters in all wheat genotypes, while in tolerant group the decrease was less in all parameters, i.e., relative water content (25.0-29.6%), membrane stability index (23.9-27.2%), total chlorophyll (41.1-42.4%) and carotenoids content (14.9-17.2%), chlorophyll stability index (30.8-36.2%) and SPAD value (26.1-28.7%) as compared to susceptible group of genotypes in which percentage change in relative water content ranged from 37.7 to 41.2%, membrane stability index (37.9-44.5%), total chlorophyll (52.6-60.2%) and carotenoids content (15.1-33.9%), chlorophyll stability index (44.6-49.9%) and SPAD value (33.5-40.9%) under combined salinity and high temperature stress treatments. However, lipid peroxidation was increased in all the genotypes with minimum percentage increase in tolerant group which ranged from 58.6 to 60.3% and maximum percentage increase in susceptible group ranged from 71.4-88.3% under combined salinity and high temperature stress treatments. Greater antioxidant enzyme activities were induced in tolerant group, i.e., peroxidase (61.8-69.8%) and superoxide dismutase activity (75.4-86.9%). Similarly, proline content (61.1-68.1%), total phenol content (59.9-71.9%), total soluble sugar (42.5-48.9%), and free amino acids (43.3-50.9%) were higher in tolerant types, in comparison to susceptible group, i.e.,, peroxidase (40.6-49.2%), superoxide dismutase activity (50.7-56.3), total soluble protein (12.2-24.4%), proline content (37.6-47.4%), total phenol content (43.8-49.0%), and free amino acids (21.1-32.2%) under combined stress condition. However, total soluble protein content was found to be decreased less in tolerant group (19.6-21.6%) then in susceptible group (36.6-28.4%) under combined stress condition. Irrespective of the genotypes, combined stress reduced the yield and yield components in all genotypes. However, the reduction was minimum in tolerant genotypes, i.e., plant height (25.8-30.6%), number of grain per ear (28.6-34.1%), test weight (29.2-31.9%), harvest index (36.7-39.0%) and yield per plant (37.6-43.6%) as compared to susceptible genotypes in which reduction in plant height ranged from 36.9 to 49.6%, number of grain per ear 44.4 to 49.4%, test weight 38.7 to 43.4%, harvest index 47.3 to 50.3% and yield per plant 53.5 to 58.3% under combined stress treatments. However, floret sterility index was increased in all the genotypes with minimum percentage increase in tolerant group which ranged from 67.0-74.1% and maximum percentage increase in susceptible group ranged from 96.3 to 101.2% under combined salinity and high temperature stress treatments. From the present investigation it was evident that tolerant genotypes, i.e., KRL-19, KRL-1-4 and HD-2733 had the potential to cope up with the adverse effect of given stress treatments and they performed well under stress condition with least reduction in physiological, yield and yield associated parameters and greater increase in biochemical parameters. Among all parameters, antioxidants enzyme activity (SOD), phenol and proline content were highly correlated with yield per plant and showed maximum induction under combined salinity and high temperature stresses. Hence, the higher percentage increase in these parameters may be the reason for the tolerance of genotypes, HD-2733, KRL-19 and KRL-1-4. Possibly, some of these indices might prove useful for improving wheat genotypes to withstand combined salinity and high temperature.ThesisItem Open Access Stability study in spring planted early maturing sugarcane clones for higher yield(Dr. Rajendra Prasad Central Agricultural University, Pusa, Samastipur (Bihar), 2018) Kumar, Praveen; Pandey, S.S.Fifteen early maturing sugarcane genotypes were planted in spring season at two different locations viz. Pusa and Madhopur for two consecutive years (2016 and 2017) to assess their stability across four environments viz. E1 – Pusa‟2016, E2 – Madhopur‟2016, E3 – Pusa‟2017 and E4 – Madhopur‟2017, in Randomized Block Design with three replications for yield and attributing traits. The genotypes showed high amount of variability present in individual (E1, E2, E3 and E4) and pooled over environments for most of the characters. Genotypes viz. CoP 16437, CoP 16436, CoP 11437 and CoP 16438 showed superiority for most of the traits including sugar and cane yield. PCV was found higher than GCV revealed environmental influence, sugar yield exhibited high heritability coupled with high GAM revealing preponderance of additive effects of gene while cane yield at harvest showed high heritability coupled with moderate GAM. Significant G × E interaction including environment (linear), linear component of G × E interaction and pooled deviation (non-linear) were significant indicating considerable genetic variability for most of the studied traits. Role of the environmental variation was observed for most of the attributes except for the number of shoots at 120 days, plant height (PH) at 240 days, single cane weight (SCW), number of milliable cane (NMC) and cane yield (CY) at harvest revealing no effect of environment or similar behaviour and response of studied genotypes. As per estimated stability parameters, six genotypes were found stable viz. CoP 16437, CoP 11438 and BO 153 stable for CY and SY in rich environment. CoP 16436 stable in poor environment for CY while in rich environment stable for SY. CoP 16438 stable in poor and rich environment for SY and CY respectively whereas CoP 11437 was stable for CY in rich environment. In AMMI analysis, G × E interaction was found highly significant for most of the traits, IPCA I and IPCA II were also found highly significant for most of the traits. IPCA I and IPCA II cummulatively contributed more than 80 percent of the total G × E interaction for all traits. All four environments E1, E2, E3 and E4 were identified as favourable for cane yield and sugar yield at harvest and fall into two groups i.e. one group (E2 and E4) and another group (E1 and E3) were preferred for most of the traits and had similar effects on genotypes. Biplot analysis revealed that genotypes CoP 16437, CoP 16436, CoP 11437, CoP 16438, BO 153 and CoP 11438 were found the most desirable and favourable across different environments. As per both the stability model the genotypes viz. CoP 16437, CoP 16436, CoP 11438, CoP 16438 and CoP 11437 were identified as stable one for yield and yield attributing traits in their respective environments.ThesisItem Open Access Stability study on basal and non-basal branching genotypes of Indian Mustard (Brassica juncea L. Czern & Coss) under different moisture regimes(Dr. Rajendra Prasad Central Agricultural University, Pusa, Samastipur (Bihar), 2018) Chandra, Khusboo; Pandey, AnilKeeping in view large acreage of Indian mustard under conserved residual moisture – rainfed condition on farmers‟ fields, the present study entitled “Stability study on basal and non-basal branching genotypes of Indian mustard (Brassica juncea L. Czern & Coss) under different moisture regimes” was undertaken. Two field experiments: Divergence study under rainfed condition (Rabi 2015-16) with 50 genotypes from 10 AICRP-R&M centres located in major crop Brassicas growing areas; Stability study (Rabi 2016-17) with 20 genotypes, under four environments viz, no irrigation under Rainout Shelter (E1), rainfed (E2 ), one irrigation : 45 DAS (E3 ) and two irrigations: 45 and 65 DAS (E4 ), conducted in Randomized Complete Block Design in three replications and evaluated for twenty – five and thirty – three morpho-physio-biochemical traits, respectively along with laboratory experiment for drought related thirteen physiological traits. 50 genotypes for investigated characters revealed significant amount of variability, categorized as 13 BB and 37 Nbb (27 M-Nbb and 10 H-Nbb) plant types. Rajendra Suphlam and NDRE7 (BB), Pusa Mahak and NRCDR-2 (M-Nbb) and Rohini (H-Nbb) reflected superiority for many traits as compared to best national check basal – branching variety Varuna. High level of variation with more number of PBB, SB-PBB, less IL-PBB, bold seeds – PBB on basal branches observed as compared to Nbb on all 13 BB genotypes. PCV higher than GCV revealed environmental influence, PBP-1, SBP-1, NS, SG, IL, HFPB, HFS, AS, AB, RL, RV and RG exhibited high h2bs and Genetic advance as percent of mean revealing preponderance of additive effects of gene. Early DFF, DPM and less IL showed negative whereas, RV, RG and SS-1 exhibited positive association of traits with GYP1. Multivariate analyses, Tocher‟s, Euclidean and PCA methods reflected maximum contribution (85.39%) of root length, HFPB and RV towards total divergence. The suggested crosses with the most divergent parents involved all branching patterns (BB, M-Nbb and H-Nbb) in 13 crosses combining 9 (Tocher‟s and Euclidean method) and more reliable in six divergent crosses between six most divergent genotypes: Rajendra Suphlam/ Divya, Rajendra Suphlam/ RH0406 and Rajendra Suphlam/ TM-2 (BB/BB); Pusa Mahak/ Divya (M-Nbb/BB); Rajendra Suphlam/ Kranti(BB/M-Nbb) and Pusa Mahak/ Kanti (M-Nbb/H-Nbb) common from Tocher‟s, Euclidean and PCA methods, were suggested for future Brassica improvement. High amount of variability individual (E1, E2, E3, E4) and pooled over environments reflected worth of studied genotypes for most of the characters. Significant G-E interaction including environment (linear), linear component of G-E interaction and pooled deviation (non-linear) were significant indicating considerable genetic variability for most of the studied traits. Role of the environmental variation was observed for most of the attributes except no influence on SD, LS, SS-1, LAI, CC, PRO, TSW and OC. Out of eleven stable genotypes, seven (NRCDR-2, TM-151, Kranti, PKRS-28, TM-128, PM-28 and RAURD-78) in poor, two (Rajendra Suphlam and KMR 10-2) in average and two (Rohini and RH8814) in rich environments. NRCDR-2 and Rajendra Suphlam, both stable genotypes for yield in poor and average environments, respectively, exhibited stability for RGR, LAI,SLW, HI and DME( in poor) and for RV, DPM, CC, PRO and BY (in rich) environments. But for HFPB and TSW, Rajendra Suphlam reflected stability in poor whereas for HFPB, NRCDR-2 expressed in rich environment. For none of the 11 stable genotypes DFFO reflected stability and needs attention for ideotype development except RH-8814 in rich environments. NRCDR-2 (M-Nbb, Variety 2007) is the most stable genotype in poor whereas popular variety Rajendra Suphlam (BB) stable in average environments can be suggested to farmers for residual moisture – rainfed condition. Suggested plant ideotype based on findings of present investigation for moisture stressed – rainfed condition would be basal – branching, semi – compact growth habit (AB) and semi – appressed (less AS) siliqua bearing one; with more than 50% PBBs accommodating SBs, less IL, lower positioning of HFS, deep tap root with more volume (RL and RV), earliness in DFF and DPM, high RGR, LAI and SLW up to early DFFO stage, bold seeds and other yield components like LS, SS-1 and DME. Yield determinants RV, RL and HFPB are most important for residual moisture – rainfed environments as reflected by their contribution towards total divergence, high significantly positive (RL, RV) and negative ( HFPB) association, regression direct effect towards yield along with high h2bs and Genetic advance as percent of mean. Overall, RV as best yield contributor also reflected stability in rich environment among stable genotypes for yield (poor and rich environments) and could be utilized for mustard varietal improvement in moisture stress.ThesisItem Open Access Characterization and micropropagation of Physalis minima L. in Bihar(DRPCAU, Pusa, Samastipur, 2018) Kumari, Anjani; Kumar, HarshA study was undertaken to characterize populations of Physalis minima L. collected from the seven locations of Bihar. A list of 47 morphological characters were used to score the variations present in the plants. Qualitative and quantitative estimation for the presence of phytochemicals was done to determine the medicinal and nutritive value of the plant. Different populations were also characterized on the basis of their molecular study using ISSR and SSR markers. The superior plant populations were identified on the basis of these studies. The micropropagation of the plants was carried out for their multiplication and maintaining superior characters and also prevent the natural hybridization. The morphological characterization based on 24 qualitative and 13 quantitative characters revealed the presence of variations within and between populations. The variations not reported before were observed, which lead to the identification of new forms of the Physalis minima L. plants in Bihar. The quantitative morphological traits were found to be significantly correlated. The cluster analysis and Principal Component Analysis successfully grouped the populations into three groups. The presence of eight phytochemicals were detected in leaves, stem, fruits and roots of the plants from all the seven populations by using thirteen qualitative tests. The quantitative estimation revealed the presence of a high amount of total soluble sugar, ascorbic acid, protein, phenols, flavonoids and alkaloids in leaves and fruits of the seven populations. The populations showed significant variations in their phytochemical constitution. The dissimilarity coefficient and Principal Component analysis based on quantitative biochemical characters clustered the populations in three groups similar to the groups identified by morphological characterization. The biochemical profile highlighted the fruit and medicinal value of the plant. Three out of the eight SSR markers used in the study failed to achieve amplification in the forty two plants of Physalis minima, suggesting a lack of transferability of these primers. A total of 50 alleles were detected using the five SSR primer pairs. The plants were effectively diversified using the SSR primer based similarity coefficients and Principal Coordinate Analysis. The plants were grouped in three clusters having the plants from different populations. A total of 574 alleles were detected using fifteen ISSR primers. The results of ISSR marker based similarity coefficient and Principal Coordinate Analysis showed corresponding results. The plants were again grouped into three clusters. The similarity coefficients and Principal Coordinate Analysis based on both the markers revealed a similar result, suggesting the applicability of both the markers in scoring the genetic variation present in the Physalis minima L. plants. A high level of genetic heterogeneity was identified in the populations based on molecular studies. The results of the three studies were found to be related suggesting that the variations in morphological and biochemical parameters are a result of the genetic variations. The Selao was identified as the superior population based on the score of biochemical characters. Since the Physalis minima L. populations are highly heterogeneous, there is a need for purification of the populations which can lead to the development of superior genotypes. The tissue culture responses were found to be independent of the effect of population. The shoot apical bud followed by nodal stem were identified as the best explant for most of the tissue culture response except swelling and callogenesis for which leaves were the most effective. MS 30 (1 mg l-1 BAP and 1 mg l-1 2, 4-D) was identified as the best medium for differentiation of multiple shoots in shoot apex and nodal stem while MS 2 (1 mg l-1 BAP) was the best for leaves. MS 7 (1 mg l-1 KIN) was invariably identified as the best medium with the highest number of shoots per explant for all the explants. The leaves were the best explant for callogenesis. The in vitro flowering and fruiting were obtained from shoot apex and nodal stem culture. MS 32 (1.0 mg l-1 BAP and 1.5 mg l-1 2, 4-D) for flowering and MS 30 for fruiting was identified as the best medium for both the cultures. R3 (MS + 1 mg l-1 NAA) was identified as the best medium for rooting. The in vitro studies led to the development of a highly efficient protocol for micropropagation of Physalis minima L.ThesisItem Open Access In vitro morphological and molecular evaluation of rice genotypes and validation of markers for salinity tolerance(DRPCAU, Pusa, Samastipur, 2018) Kumari, Rima; Kumar, HarshA set of thirty rice varieties including two tolerant (Pokkali and CSR-36) and two susceptible (IR-29 and IR-64) checks were screened for assessment of their salt tolerance at early seedling stage on the basis of in vitro seed germination and seedling growth at different salinity levels (0, 4, 8 and 16 dS m-1) created by application of salt mixture consisting of NaCl, CaCl2, Na2SO4 in 7:2:1 ratio. Seed germination and seedling growth were adversely affected with the increase of salt concentrations. The salt tolerance index (STI) of the rice varieties was calculated on the basis of seed germination, seedling shoot and root dry weights at different levels of salt stresses. Phenotypic grouping of the thirty rice varieties for their salt tolerance level on the basis of overall salinity tolerance indices across in vitro seed germination and seedling growth (shoot and root dry weight) under salt stress clearly reflected that seventeen varieties, namely, Pokkali, CSR-36, Mandakini, Kranthi, Jyothi, Bardhan, Pusa Sugandh-2, Duna Sankhi, Sanwal Basmati, Ratnagiri-4, Shanthi, Rajendra Dhan-102, Sahbhagi Dhan, Vaisak, Annada, Badami and Jyotrirmayee were highly tolerant to salt stress. Among the remaining entries, Pusa-834, Sarsa, Govind, Khira, Pusa Sugandh-5, MTU-7029 and Saraswathi were found to be moderately salt tolerant, whereas varieties IR-29, IR-64, Daya, Kalinga-3, Golaka and Shatabdi were highly susceptible to salt stress. Out of thirty rice varieties, eighteen rice varieties, namely, Pokkali, CSR-36, IR-29, IR-64, Mandakini, Pusa Sugandh-2, Saraswathi, Ratnagiri-4, Rajendra Dhan-102, Sahbhagi Dhan, Badami, Sarsa, Jyotrirmayee, MTU-7029, Golaka, Daya, Vaisak, and Shatabdi were further screened with the help of ISSR, SSR, salt stress responsive candidate genes and EST primer pairs for the purpose of their molecular profiling in relation to their salinity tolerance. Genetic profiling of entries with a panel of 14 ISSR markers generated altogether 483 allelic variants including 236 shared and 247 unique alleles with an average of 34.50 alleles per primer, revealing ample extent of genetic differentiation and divergence amongst the entries. Among these markers, 811, 814, 815, 823, 834, 836, 840, 841, 842, 872 were found to be highly polymorphic and informative on the basis of their PIC and PP values. However, using a panel of salt stress response related 24 SSR primer pairs, altogether 205 allelic variants including 114 shared and 91 unique alleles were detected with an average of 8.54 alleles per primer due to length variation of simple sequence repeats. Simple sequence repeat loci with di-nucleotide and tri-nucleotide repeat motifs detected greater number of alleles than the repeat loci with tetra-nucleotide and complex repeat motifs. Additionally, the simple sequence repeat loci with CT, GT, AT, AG and AC di-nucleotide repeat motifs detected greater number of alleles. Contrarily, the loci with GA and CA di-nucleotide repeat motifs appeared to detect relatively lesser number of alleles. Considering the number of alleles generated in conjunction with the level of polymorphism detected in the present study, the primers RM 302, RM 8094, RM 10665, RM 10694, RM 10748 and RM 10825 appeared to be highly polymorphic and comparatively more informative primers. Six SSR primer pairs, namely, RM 140, RM 1287, RM 3412, RM 10745, RM 10764 and RM 10772 were validated on the basis of their efficiency to distinguish salt tolerant varieties from susceptible varieties. These six primers can be utilized for the purpose of genetic differentiation and discrimination in relation to salt stress responsiveness of the rice genotypes. Similarly, microsatellite containing salt stress responsive candidate gene (OsHKT1;5, SNAC1, CDMK, CCC, SHMT1 and SHMT2) and microsatellite lacking salt stress responsive candidate gene (OsHKT1;1, OsHKT1;3, OsHKT2;3 and OsHKT2;4) specific markers based genetic profiling allowed unambiguous discrimination of salt stress responsive and tolerant entries, validating their utility for the purpose of differentiation and discrimination of salt stress sensitive and tolerant varieties. Principal coordinate analysis completely supported the results obtained from hierarchical classification of the varieties. Using a panel of eight salt stress responsive EST-contigs based markers, monomorphic bands amongst all the 18 varieties were recorded for six markers, namely, Contig2 (Ferritin superfamily), Contig54 (Plant peroxidase superfamily), Contig138 (ATPase expression protein), Contig314 (Exonuclease), Contig545 (Major latex protein) and Contig633 (Protein kinase), revealing genetic similarity with respect to primer binding sites and molecular size of targeted genomic regions. Remaining two markers specific to Ferritin superfamily (Contig43) and Microtubule associated protein (Contig215) genes revealed genetic polymorphism in the form of presence or absence of bands. Thus, combining in vitro morphological and molecular assessment, seventeen varieties were considered as salt tolerant varieties which can be used as parental donor in rice breeding programme to develop salt tolerant rice varieties.ThesisItem Open Access Studies on spot blotch of wheat caused by Bipolaris Sorokiniana (SACC.) Shoem(DRPCAU, Pusa, Samastipur, 2018) Kumar, Paritosh; Rai, R. C.Studies on spot blotch of wheat (Triticum aestivum L.) was carried out to ascertain the status of spot blotch disease of wheat in Bihar, to find out the variability of pathogen in Bihar and to find out a suitable culture media for growth and sporulation of the pathogen. Attempts were also made to estimate the yield losses; and to manage the disease by agronomical manipulations and through fungicides and botanicals. Bipolaris sorokiniana was mainly responsible for causing spot blotch of wheat in Bihar. The disease appeared 5 to 8 weeks after wheat sowing and the symptoms advanced after heading and spreaded very fast. The dark brown necrotic spots (boat shaped) occured on the leaves, crowns and stems with or without yellow halo around these. Lesions on the leaves in the start were few mm in size and extended as elongated dark brown spots and later on coalesced resulting into blight. In severe cases, at flowering stage 60 to 80 per cent leaf areas was turned into necrotic lesions. Intensity of spotting increased with the increase in the age of plants and reached maximum between boot to maturity stage of the crop. Minute spots were also observed on leaf sheaths, nodes, internodes and the glumes. The conidiophores of Bipolaris sorokiniana were light olivaceous brown in colour and emerged both singly or in a group of 2 to 6 from stomatal opening or directly through the epidermal cells of the host, bearing conidia on well defined geniculations and measured 81 to 193 μm long (average 155.20 μm), 6 to 8 μm wide (average 6.70 μm) having 2 to 9 septa (average 6.40). Conidia were formed singly at the tip of conidiophores and were dark olivaceous brown in colour, straight to slightly curved, widest at or near the middle, tapering towards both the abruptly rounded ends and measured 39 to 112 μm long (average 73.90 μm), 13 to 21 μm wide (average 16.70 μm) having 2 to 6 septa (average 3.70). Six groups of B. sorokiniana could be differentiated on the basis of colony colour, colony characteristics and colony diameter establishing the variability in Bipolaris sorokiniana. Remarkable variation in length, width, number of septa in spore and shape of spore were present. Oat meal agar medium and oat meal medium were best suited medium for radial growth and biomass production of Bipolaris sorokiniana, respectively. However, potato dextrose agar was best suited for sporulation of Bipolaris sorokiniana. Yield loss due to spot blotch varied between 7 to 30 per cent and loss in 1000-grain weight between 3 to 23 per cent, depending upon the levels of disease. Delayed sowing favoured incidence of spot blotch irrespective of nitrogen level and more disease developed at higher nitrogen level in all the three date of sowing. Higher levels of nitrogen at all the three date of sowing increased yield and 1000-grain weight and with delay in sowing, yield and 1000-grain weight decreased significantly at all the three nitrogen level. Propiconazole, Hexaconazole and Difenaconazole + Propiconazole at 250 ppm completely inhibited the mycelial growth of B. sorokiniana. Carbendazim and Copper oxychloride also produced cent per cent inhibition but at 500 and 1000 ppm, respectively. Mancozeb could not produce complete inhibition even at 1000 ppm. Spot blotch of wheat could be effectively and economically controlled by two foliar sprays of either Propiconazole or Hexaconazole @ 0.1 per cent at fifteen days interval but on the basis of cost- benefit ratio Hexaconazole had an edge over Propiconazole. Among botanicals eucalyptus leaf extract was highly effective in inhibiting the growth of B. sorokiniana. Garlic clove extract was second best plant extract in inhibiting the growth of B. sorokiniana. All the six plant extracts (garlic clove, eucalyptus leaf, neem leaf, onion bulb, ginger rhizome and black cumin) could significantly reduce PDI when compared with untreated check. Two sprays of garlic clove and eucalyptus leaf extracts @ five per cent were equally effective in reducing disease to 50 per cent and increasing grain yield. Two spray of Propiconazole (0.1 %) at 15 days interval under timely sown condition was best in controlling spot blotch of wheat with significant increase in yield over unsprayed crop. With the delayed sowing (26th December) there was significant decrease in disease control with significantly less yield. Hexaconazole was equally effective and on the basis of cost- benefit ratio, had an edge over Propiconazole.ThesisItem Open Access Biotechnological development of Panama wilt resistance in local cultivars of banana(RAJENDRA AGRICULTURAL UNIVERSITY, BIHAR PUSA (SAMASTIPUR), 2017) Kumari, Anita; Kumar, HarshBiotechnological development of resistance to Panama wilt, the most important disease caused by Fusarium oxysporum f.sp. cubense, was explored in four important local cultivars of banana namely Robusta, Alpan, Malbhog and Kothia employing tissue culture studies and genetic transformation. Using tissue culture, efficient micropropagation protocol and disease resistance utilizing somaclonal variation were developed. Biological control and management of the disease was done utilizing endophytic association of nonpathogenic strain of Fusarium oxysporum. The culture of shoot apices and male flower buds resulted in their establishment and swelling, and callogenesis, caulogenesis, differentiation of shoots and roots from them. Elongation of existing shoots was observed from shoot apices and somatic embryogenesis from floral buds only. The best medium for caulogenesis was BM19 (MS+1.14 µM IAA+19.97 µM BAP) for cultured shoot apices and medium BM8 (MS+22.19 µM BAP) for cultured male flower buds. The best medium for somatic embryogenesis was BM20 (MS+18.10 µM 2,4-D+5.37 µM NAA+ 5.71 µM IAA+ 1mg Biotin). The cultivar Robusta was the best for all tissue culture responses except somatic embryogenesis followed by either cultivar Alpan or cultivar Malbhog. The cultivar Kothia showed the least tissue culture responses. However, the best response for somatic embryogenesis was observed in cultivar Kothia followed by cultivars Malbhog, Alpan and Robusta respectively. The subculture of in vitro developed multiple shoots on medium BM19 resulted in cent percent shoot multiplication with a high multiplication rate. An efficient micropropagation protocol was developed for banana cultivars. The Panama wilt resistance was developed in two susceptible cultivars Alpan and Malbhog exploiting somaclonal variation. Endophytic nonpathogenic Fusarium oxysporum strains were found effective in biocontrol of wilt disease in banana. Genetic transformation of callus and regenerating shoots was done using GUS and eGFP marker gene constructs and it was confirmed using histochemical GUS assay and fluorescence microscopy respectively, and by molecular validation using specific SSR primer pairs. Thus, the work established a suitable protocol of the genetic transformation, which can be used for the development of Panama wilt resistance in local cultivars of bananas.ThesisItem Open Access Effect of summer legumes on growth and yield of succeeding direct-seeded rice (Oryza sativa L.) under different nitrogen levels(Dr. Rajendra Prasad Central Agricultural University, Pusa, Samastipur, 2018) Choubey, Anup Kumar; Sinha, K. K.A field experiment was carried out during the two consecutive kharif season of 2014 and 2015 at research farm of Tirhut College of Agriculture, Dholi of Dr. Rajendra Prasad Central Agricultural University, Pusa, Samastipur, Bihar to study the effect of summer legumes on growth and yield of succeeding direct-seeded rice (Oryza sativa L.) under different nitrogen levels. The experiment was laid out in split-plot design and replicated thrice. The treatments consisted of three summer legumes viz. dhaincha, mungbean, urdbean and summer fallow was kept in main plot and four nitrogen levels viz. 0% RDN (recommended dose of nitrogen), 50% RDN, 100% RDN and 150% RDN in sub-plot. All the treatments received an uniform application of 60 kg P2O5, 40 kg K2O and 25 kg ZnSO4 per hectare as basal dressing. Nitrogen was applied in three equal splits i.e. 1/3rd as basal, 1/3rd at the active tillering stage and rest 1/3rd at panicle initiation stage. The crop was sown in rows 20 cm apart using the seed rate of 30 kg/ha. The soil of the experimentation plot was sandy loam in texture and low in OC, available Nitrogen, Potassium and medium Phosphorus. Rice crop sown after incorporation of summer legumes significantly influenced the growth, yield attributes and yield during both the years. Significantly higher plant height (85.15 & 86.03 cm), number of tillers (291.25 and 302.50 m-2), dry matter production (872.19 and 891.36 g m-2) and leaf area index (4.00 and 4.14) were recorded when crop was sown after incorporation of dhaincha than summer fallow. Rice crop was sown after incorporation of dhaincha recorded maximum number of panicles (275.00 and 285.12 m-2), panicle length, number of spikelets/panicle, number of fertile spikelets/panicle being at par with preceding mungbean and both recorded significantly higher value over summer fallow. The crop sown after incorporation of dhaincha produced significantly higher grain yield (38.74 & 39.59 q ha-1) than mungbean, urdbean and summer fallow. Nutrient uptake, net return (`32813 & 38960 ha-1) and B: C ratio (1.10 & 1.36) of rice were also highest in dhaincha incorporated plot which was significantly higher over mungbean, urdbean and summer fallow. Similarly, incorporation of mungbean and urdbean statistically at par in respect to yield indices, grain yield, net return and B: C ratio and both recorded significantly higher values than summer fallow. Application of 100% RDN in dhaincha incorporated plot recorded significantly higher grain yield, net return and B: C ratio than the application of 150% RDN in rice followed by urdbean and summer fallow. Application of 150 per cent recommended dose of nitrogen registered maximum plant height (87.90 and 88.88 cm), number of tillers (309.58 and 318.33 m-2), dry matter production (877.78 and 896.95 g m-2) and leaf area index (4.20 and 4.37) which was statistically at par with 100% RDN and booth significantly scored over 50% RDN and control. The maximum yield attributing parameters, grain yield (40.07 & 41.46 q ha-1), net return (` 34259 & 39880 ha-1) and B: C ratio (1.12 & 1.35) were also recorded at 150% RDN which was found at par with 100% RDN and both significantly scored over lower levels of nitrogen. The N, P and K uptake by rice increased significantly with increasing levels of nitrogen and recorded significantly higher uptake at 150% RDN than lower levels of nitrogen.
