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ThesisItem Unknown Identification of noval donors for resistance against Tilletia indica causing Karnal bunt from wild and related species of wheat(Punjab Agricultural University, Ludhiana, 2020) George, Anju Grace; Vineet KumarKarnal bunt (KB) of wheat caused by the fungus Tilletia indica is an internationally quarantine-significant disease of wheat. KB can be managed in a better way by exploiting genetic resistance in wild and related species of wheat. A population of 490 lines derived from progenitor species of wheat such as Aegilops tauschii and Triticum spp. (T. monococcum, T. boeoticum, T. dicoccoides), non-progenitor species Triticum araraticum and Aegilops triuncialis and triticale introgressions with wheat was utilized in this study. This study was conducted at the Experimental farm and laboratories, Department of Plant Breeding & Genetics, PAU Ludhiana during rabi seasons of 2018-19 and 2019-20. Among the progenitor and non-progenitor species and their bread wheat introgressions, Triticum araraticum 4747 /WHD 943 // bread wheat showed the highest level of KB resistance. This was followed by PDW 233 / Aegilops tauschii 14119// bread wheat. Among the triticale lines with diverse genetic background and their introgressions with bread wheat, TL 2908/2*CS(S)// DBW17 showed the highest level of KB resistance followed by T 3021/2* CS (S) // PBW 550. These lines were considered as promising donors for KB resistance and can be utilized for further breeding disease resistant varieties. Expressed Sequence Tag (EST) based markers specific for chromosome arms of rye were used to determine rye chromosomes associated with the KB resistant triticale x bread wheat lines. The introgressed regions of rye chromosome 4RL, 4RS and 7RL might have contributed to KB resistance in the phenotypically resistant lines.ThesisItem Restricted Bio-management of chickpea wilt incited by Fusarium oxysporum f.sp. ciceri (Padwick) Synd. & Hans.(Punjab Agricultural University, Ludhiana, 2020) Arpanpreet Singh; Narinder SinghFusarium wilt of chickpea caused by Fusarium oxysporum f.sp. ciceri is a serious disease in chickpea growing areas of Punjab. Rhizosphere soil samples were collected from chickpea fields of Ludhiana, Gurdaspur, Mansa, Moga and Bathinda regions and analysed for antagonistic microflora. Five isolates of Trichoderma spp. and three isolates of Pseudomonas spp. were isolated and screened in vitro for antagonistic activity against Fusarium oxysporum f.sp. ciceri causing wilt of chickpea. Th5 isolate of Trichoderma viride and Pf2 isolate of Pseudomonas aeruginosa showing maximum inhibition of 73.11 and 67.63 per cent against Fusarium oxysporum f.sp. ciceri in dual culture assay were selected. Th5 isolate also showed highest chitinase activity 8.85 unit/ml and Pf2 showed highest siderophore activity with the production of 29.07 mm diameter of orange to yellow coloured halo. Efficacy of talc based bioformulations of Trichoderma viride and Pseudomonas aeruginosa isolates applied as seed, soil and seed + soil treatment was seen against Fusarium wilt of chickpea under field and pothouse conditions. Maximum disease inhibition was observed in chemical treatment (Carbendazim) with 88.95, 88.12 and 88.72 per cent in Ludhiana, Gurdaspur and Pothouse trial, respectively, followed by the Trichoderma viride (Th5) seed + soil treatment with 81.84, 85.69 and 83.63 per cent disease inhibition in Ludhiana, Gurdaspur and Pothouse trial, respectively. Maximum growth promotion was also recorded in chemical treatment (Carbendazim) i.e. shoot length, root length and ultimately increased the yield of chickpea under both field and pothouse conditions followed by the isolate Trichoderma viride (Th5) seed + soil treatment. The rhizospheric competence of the antagonists showed that enough population density of antagonists was present throughout the crop period to control the pathogen both under both field and pothouse conditions. Maximum shelf life of talc-based powder bioformulation of all the antagonists was recorded in low temperature conditions (0°C) when packaged in polythene bags.ThesisItem Open Access Characterization of Tilletia barclayana causing kernel smut of rice and its management(Punjab Agricultural University, Ludhiana, 2020) Babanpreet Singh; Anju BalaKernel smut of rice caused by Tilletia barclayana is a designated seed borne disease under Indian minimum seed certification standards. The pathogen converts the grains into black powdery mass resulting considerable loses in the yield as well as quality of the produce. The short duration rice varieties are more susceptible to kernel smut as compared to the other varieties. The present studies aimed at characterization of the populations of T. barclayana isolates obtained from different varieties under cultivation in Punjab from different agro-climatic regions and to devise management of this disease. A total of thirty three isolates of T. barclayana were obtained from thirteen districts and eight different varieties for morphological and pathological characterization. Based on the morphological and cultural characteristics, the isolates were grouped into 8 clusters, from each cluster representative isolates were selected for their pathological characterization under field conditions. Fourteen isolates were used for pathological characterization using spray inoculation technique at fifty per cent flower opening and by syringe inoculation technique at late boot stage. Multivariate cluster analysis of the pathogenicity data grouped the isolates into five and three clusters by spray and syringe inoculation method respectively. Isolate Tb 24 obtained from variety PR 124 from district Ropar was the most virulent isolate of T. barclayana whereas isolate Tb 31 obtained from variety PR 121 from district Tarantaran was the least virulent among the isolates under study. The germinated teliospores obtained from weed hosts, Dichanthium annulatum, Cyanodon dectylon were inoculated on variety PR 114 by spray inoculation method to study the cross infectivity of T. barclayana from weeds to rice. Neither of the isolate from any weed hosts were able to produce disease symptoms on rice. Six different fungicides including Hexaconazole 75% WG, Propicinazole 13.9% + Difenconazole 13.9% EC, Hexaconazole 4% + Zineb 68%, Tebuconazole 50% + Trifloxystrobin 25% WG, Azoxystrobin 23% SC, Propiconazole 25% were evaluated against T. barclayana under lab conditions and for disease control under the field conditions. Minimum ED 50 (3.57 ppm) and ED 90 (37.11 ppm) values were recorded for Hexaconazole 75% WG. Maximum disease control under field conditions was achieved in plots where two sprays of combination fungicide Hexaconazole 4% + Zineb 68% was given. The results of two sprays of Propiconazole 25% EC and combination of Tebuconazole 50% + Trifloxystrobin 25% WG @ 0.1 % and 0.04% respectively were equally effective for the management of disease under field conditions Fifty advanced breeding lines of rice were screened for disease reaction under natural conditions. Disease incidence and coefficient of infection was categorized into different response of disease reaction on the fifty advanced breeding lines under study at Ludhiana and Kapurthala. At PAU Ludhiana fourty six lines exhibited resistance response and four lines exhibited highly resistance response. At RRS Kapurthala 41 lines exhibited resistance response, one of the line was susceptible and seven lines were moderately susceptible to kernel smut under natural conditions. The lines showing resistance response can further be evaluated under artificial inoculation conditions in order to obtain resistance sources against the pathogen. The most virulent pathogen isolate Tb-24 may be used for artificial screening of lines against T. barclayana.ThesisItem Open Access Biocontrol of fusarial wilt of bottle gourd(Punjab Agricultural University, Ludhiana, 2019) Ruchika, Ruchika; Narinder SinghThe wilt of bottle gourd caused by Fusarium oxysporum is a serious disease in bottle gourd growing areas of Punjab. Rhizosphere soil samples were collected from bottle gourd fields of Ludhiana, Kapurthala and Bathinda districts of Punjab in order to analyze the antagonistic microflora. Five isolates of Trichoderma and three isolates of Pseudomonas were isolated and screened in vitro for their antagonistic activity against Fusarium oxysporum. Among the various isolates, two isolates of Trichoderma (T1 and T2) and one isolate of Pseudomonas (P1) showed good inhibition against Fusarium oxysporum under lab conditions. Trichoderma isolate T2 showed maximum inhibition of Fusarium oxysporum (76.33 % and 65.83%) in dual culture and volatile assay, respectively. On Molecular characterization the selected isolates T1 and T2 were identified as Trichoderma asperellum and isolate P1 was identified as Pseudomonas aeruginosa. The selected isolates were then applied as seed, soil and seed + soil treatments using 10g and 15g formulation of each antagonist to check their efficacy against Fusarium oxysporum in two different seasons (March and June) under field conditions. Maximum germination (91.67% and 87.50%) and maximum disease inhibition (92.31% and 86.67%) was found in case of chemical treatment followed by Trichoderma asperellum isolate T2 with a germination of 87.50 per cent and a disease reduction of 84.62 and 80.00 per cent in March and June, respectively. Maximum yield (235.99q/acre and 229.00q/acre) along with maximum total length (5.43 m and 4.83 m) was obtained in case of Trichoderma asperellum isolate T2 in March and June, respectively due to disease control and the plant growth promoting behaviour of the antagonist. The results of rhizospheric competence showed that enough propagules of the antagonists were present throughout the growing season to effectively manage the wilt pathogen. Talc based bioformulation of the selected antagonists showed maximum shelf life on storing at 0°C and packing in polythene bags.ThesisItem Open Access Components of partial resistance against Neck Blast (Pyricularia Oryzae) in basmati rice(Punjab Agricultural University, Ludhiana, 2020) Sohal, Manarshroop KaurThe neck blast disease caused by Pyricularia oryzae was highly prevalent throughout the Punjab state with disease incidence ranging from 5.6 to 52.4 per cent on various basmati varieties. Highest disease incidence was observed in Mahal Kalan village of Barnala district. Amongst varieties, Pusa Basmati 1121 had highest value of disease incidence followed by Pusa Basmati 1401, Pusa Basmati 1509 and CSR 30. Significant variability amongst the neck blast isolates of the pathogen was observed with respect to various morphological and cultural characters viz., colony colour, colony texture, colony margin, mean radial growth (mm), spore size and sporulation intensity. The average sporulation intensity of the isolates on Oatmeal agar medium was 7.55×106 conidia /ml which was higher than the average sporulation intensity of isolates on Paspalum grass medium i.e. 6.97×106 conidia /ml. Three isolates namely NB 55, NB 57 and NB 60 were categorized as fast growing isolates with radial growth of >8mm per day. Various architectural features viz., flag leaf angle, flag leaf length, flag leaf width, panicle axis, panicle diameter, degree of exertion of panicle, culm length, 50% flowering and node aging were recorded in order to find correlation with various disease parameters viz., lesion length, panicle blast severity, area under disease progress curve (AUDPC), incubation period, disease incidence (%) and apparent rate of infection (R). Overall, flag leaf angle, panicle axis and node ageing emerged out to be three important components of partial resistance. Forty five basmati rice genotypes were screened under both artificial and natural conditions against the disease. Four genotypes i.e. Tetep, INGR 15001, INGR 15002 and Pusa Basmati 1637 were found to be moderately resistant to most virulent isolate NB 7 with susceptibility index was below 3 and they could be used as donors in breeding for disease resistance.ThesisItem Restricted Characterization of Uromyces viciae-fabae (PERS.) J. SCHRÖT causing lentil rust and molecular tagging for its resistance(Punjab Agricultural University, Ludhiana, 2020) Jatinder Singh; Sirari, AsmitaThe population structure of rust isolates infecting lentil and pea in different districts of Punjab was revisited on the basis of cross infectivity test and molecular characterization. Both lentil and pea rust isolates were found non-cross infective at different stages of plant growth. The ITS region of ~700 bp, amplified from DNA of these isolates showed 97 to 99.24% sequence homology with Uromyces viciae-fabae with more than 90% query cover. The sequences were then submitted and accession numbers were provided by GenBank. Further, analysis showed less genetic variation among these isolates. Genetically diverse microsperma and macrosperma lentil lines could not differentiate the rust populations prevalent at Gurdaspur and Ludhiana. Further, recombinant inbred lines (RILs) developed from cross between rust resistant (FLIP-2004-7L) and susceptible (L-9-12) cultivars were phenotyped against lentil rust for genetic analysis and molecular mapping. Genetic analysis indicated the role of major gene in resistance. The phenotypic data was also used to identify molecular markers associated with rust resistance. Out of 389 SSR markers, available in public domain, two markers, namely, LcSSR 440 and LcSSR 606 were found to be linked with rust resistance at 8.3 and 8.1cM, respectively. Availability of high-density linkage maps and genome sequence information in lentil can be helpful to extend the present study in identifying candidate resistance genes against U. viciae-fabaeThesisItem Restricted BIOCHEMICAL AND HISTOPATHOLOGICAL STUDIES IN BARLEY FOR COVERED SMUT (Ustilago hordei) RESISTANCE(Punjab Agricultural University, Ludhiana, 2019) Jashandeep Singh; Vineet KumarCovered smut caused by Ustilago hordei is an important disease of barley in India. In present study, biochemical basis of resistance, histopathological & ultrastructural changes and identification of sources of resistance to U. hordei were undertaken. The biochemical basis of resistance was studied on a set of three barley cultivars viz. PL 426 (resistant), PL 891 (moderately susceptible) and VJM 201 (susceptible). Maximum level of total phenols (0.64 units min-1 g-1 fresh weight) and activity of peroxidase (POD) (86.70 units min-1 g-1 fresh weight) was recorded in PL 426 whereas maximum activity of polyphenoloxidase (PPO) (89.30 units min-1 g-1 fresh weight) and phenylalanine ammonia lyase (PAL) (1.39 units min-1 g-1 fresh weight) was recorded in PL 426 and PL 891, respectively. The aqueous extract of seedlings of PL 426 (resistant variety) inhibited 98% of teliospores at 100% concentration. Ultrastructural comparison of compatible and incompatible Hordeum vulgare-Ustilago hordei, host-pathogen interaction done with scanning electron microscope showed germinated teliospores and trailing hyphae all over the host surface in the compatible interaction. Whereas, in incompatible interaction the teliospores were intact and extracellular polysaccharides were present completely adjacent to teliospores which might be involved in hindering the germination of teliospores. Eighty three germplasm accessions comprising of 74 germplasm lines and 9 varieties were evaluated against covered smut disease. Among the 9 varieties, 5 varieties namely HBL 113, HBL 391, HBL 316, DWRUB 92 and DWRUB 123 were found to be completely free from covered smut disease under field conditions at two locations viz. Ludhiana and Faridkot. PL 172 and PL 891 were categorized as moderately resistant and moderately susceptible, respectively. Among 74 germplasm lines tested at both locations, 41 showed high level of resistance and were found free from disease. BL 1562, BL 1656, BL 1627 and BL 1562, BL 1652 lines were found to be resistant at PAU, Ludhiana and RRS, Faridkot, respectively. 17 germplasm lines showed moderate resistance against covered smut.ThesisItem Open Access Etiology of charcoal rot of maize caused by Macrophomina phaseolina (Tassi.) Goid. in Punjab(Punjab Agricultural University, Ludhiana, 2019) Ahmadi, Khalilullah; Harleen KaurFive different stages (55, 65, 75, 85 and 95 days old plants) of three maize hybrids viz. PMH 10, DKC 9108 and PMH 2 were evaluated for determination of most susceptible growth stage against charcoal rot during spring season. The data revealed that 75 days old plant was the most susceptible growth stage for charcoal rot. At this particular stage, mean disease severity (60.83 %) was maximum, resulting in minimum mean grain yield (55.27 q/ha).This stage also coincides with tasseling and silking stage of the plant. The effect of five different sowing dates on disease severity of charcoal rot of five maize hybrids during spring 2018 and 2019 revealed that crop sown on 20th January suffered minimum disease severity (40.91 and 36.3 %) resulting in maximum grain yield (74.27 and 77.89 q/ha) during spring 2018 and 2019 respectively. The plant height and cob height was found maximum on this date. As sowing date was delayed, severity of charcoal rot increased, thus decreasing grain yield, plant height and cob height. Among meteorological factors, disease severity was found positively correlated with mean temperature (+0.98) and negatively correlated with mean relative humidity (-0.92) and total rainfall (-0.90). The effect of three irrigation schedules (additional irrigation at tasseling and silking stage, local practice and stress at tasseling and silking stage) on the development of charcoal rot showed that application of an additional irrigation at tasseling and silking stage had least mean disease severity (36.28%) compared to other irrigation schedules and resulted in highest mean grain yield (72.61 q/ha). Water stress at tasseling and silking stage of the crop aggravated the disease development. Fifty four maize germplasm lines were evaluated artificially against charcoal rot, out of which five germplasm lines (PML-81, LTY 9808, JCY 13-1-1, SNL 172470 and SNL 172475) were found resistant, thirty moderately resistant, seventeen moderately susceptible and two of them were found highly susceptible. The maize hybrids sown in month of January and February coupled with additional irrigation at tasseling and silking stage resulted in low disease severity of charcoal rot, thus maximizing the grain yield. The germplasm lines found resistant after artificial screening of charcoal rot at the most susceptible growth stage (75 - 85 days old plants) during spring season could be successfully utilized in future breeding programme for maize improvement.ThesisItem Restricted Integrated management of Meloidogyne incognita and Rhizoctonia bataticola in root rot disease complex in mungbean(Punjab Agricultural University, Ludhiana, 2019) Rohit Kumar; Dhillon, Narpinderjit KaurMungbean crop is attacked by number of soil and air borne pathogens. Of the different soil borne pathogens; root-knot nematode (Meloidogyne spp.) is an important pathogen affecting growth and production of mungbean. Another soil and seed borne pathogen, Rhizoctonia bataticola causes root rot disease of mungbean. Meloidogyne incognita and R. batalicola being soil borne organisms may affect each other activity besides influencing the responses in their common host mungbean. Studies were conducted on the build-up of root knot nematode in nine different genotypes in summer and kharif season. The reaction of different genotypes was varied in two seasons. Eight of the nine genotypes were found to support multiplication of root knot nematode while decrease in nematode population was observed in only one genotype ML 2056. M. incognita populations in soil were observed to be significantly higher in kharif season in four genotypes viz., SML 1906, TMB 37, SML 1827, SML 1900 in comparison to summer season while in the other five genotypes there was no significant difference in the nematode population in two seasons. The interaction of M. incognita and R. bataticola revealed that higher incidence of root rot disease and maximum decrease in plant growth parameters was recorded in simultaneous inoculation of Meloidogyne incognita and R. bataticola. Of the one hundred two genotypes screened against M. incognita in summer and kharif mungbean; only ten genotypes were found to be moderately resistant. Of the eighty eight mungbean genotypes screened against Rhizoctonia bataticola only ten were moderately resistant. In multiple disease resistance studies, four genotypes which were moderately resistant against both pathogens were evaluated. Breakdown of resistance was observed in ML2581 and ML2560, while ML 2056 showed moderately susceptible reaction and ML2607 was observed to be moderately resistant against both pathogens. Integrated management of root knot nematode and R. bataticola in mungbean revealed that combined application of Bionematon@4l/ha + neem cake@1 t/ha + Captan (seed Treatment) was more effective than the individual application of chemical, bioformulation or cakes.
