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ThesisItem Open Access Investigations on endophytic bacteria for management of downy mildew caused by Sclerospora graminicola in pearl millet(CCSHAU, Hisar, 2019-12) Sangwan, Pooja; Kushal RajPearl millet is a valuable staple food in semi-arid regions of the world. Pearl millet is affected by many diseases among them, downy mildew incited by oomycetes pathogen Sclerospora graminicola (Sacc.) Schroet is the most widespread and destructive disease of pearl millet. Endophytes play an important role in metabolism and physiology of the host plants. They provide multifarious benefits such as growth promotion, suppression of pathogens and nitrogen fixation. In the present study, roots and stubbles were collected from sick plot area of Bajra Pathology after the harvest of the pearl millet crop during Kharif 2016 and 2017. A total of 30 pearl millet root bacterial endophytes (PMRBE1- PMRBE 30) were retrieved from crushed roots of pearl millet streaked on Nutrient agar plates. Ten endophytic bacterial isolates were retrieved from roots of kharif 2016 crop and other twenty isolates from roots of kharif 2017 crop. All endophytic bacterial isolates were assessed for biocontrol activities. Amongst 30 pearl millet root bacterial endophytes, sixteen isolates were positive for siderophore production, three isolates viz. PMRBE6, PMRBE25 and PMRBE28 were found positive for HCN production, 30% of the bacterial endophytes showed good growth on ACC supplemented plates and cellulolytic activity was found in bacterial endophyte PMRBE18. The isolates PMRBE1, PMRBE4, PMRBE5, PMRBE6, PMRBE7 PMRBE17, PMRBE18, PMRBE20, PMRBE23 and PMRBE24 which showed maximum germination, seedling vigour index I & II and germination speed of three pearl millet cultivars HHB226, HHB67 Imp and 7042 S under in vitro conditions were selected for seed treatment as well as for foliar spray under screen house and field conditions. Amongst various pearl millet root bacterial endophytes, maximum germination, minimum downy mildew incidence at 30 and 60 DAS, better growth and yield of pearl millet cultivars were found on treatment with isolate PMRBE6, PMRBE18 and PMRBE23. On the basis of biochemical characterization and partial 16S rRNA sequencing pearl millet root bacterial endophytic isolates PMRBE6, PMRBE18 and PMRBE23 were identified as Bacillus subtilis strain PD4 (Accession no. MN400209), Serratia sp. and Alcaligenes faecalis strain HX2016003 (Accession no. MN400208). Promising root endophytes may be used for IPM of downy mildew in pearl millet after field trials.ThesisItem Open Access Variability, epidemiology and management of Tilletia indica Mitra, the incitant of Karnal bunt of wheat(CCSHAU, Hisar, 2019-11) Mamta; Rajender SinghKarnal bunt disease of wheat continues to be a potential threat to production and trade of wheat. The present study was planned to study cultural, morphological, molecular and pathogenic variability in Tilletia indica, epidemiology and management of this disease. On the basis of cultural and morphological characteristic, the largest teliospore size was observed in Bhiwani isolate, whereas Kapurthala isolate was having the smallest size. Different Karnal bunt isolates colony colour were white, yellowish white, creamy, creamish yellow and creamish white whereas, culture colony growth pattern of isolates were erect, compact, clustering, crustose, appressed, spreading and leathery. Maximum teliospore germination, primary sporidia formation and secondary sporidia production was recorded at 18°C in Pantnagar isolate. Best teliospore germination occurred at pH 6.0 followed by pH 5.0 Maximum coefficient of infection was expressed in HD2967 followed by DPW 621-50. Pantnagar isolate exhibited maximum mean coefficient of infection followed by Bhatinda and the least was observed in Mahendergarh isolate. Pantnagar isolate was most virulent as well as had maximum sporulation. Karnal bunt resistant reaction in three differentials namely HD 29, HD 30 and Aldan were noted. In molecular variability twelve molecular markers were tested against 10 Karnal bunt isolates and characterized into 3 major groups. Karnal bunt incidence had significant correlation with maximum temperature, rainy day and total rainfall during earhead emerging stage Two hundred seventy nine genotypes showed less than 5 coefficient of infection against Karnal bunt thereby expressing resistant reaction and 221 genotypes were susceptible having more than 5 per cent coefficient of infection under artificial inoculation condition. Maximum disease control was obtained by propiconazole 25% EC (0.1%) application, followed by combination of propiconazole 25% EC (0.05%), neem seed kernel and Trichoderma harzianum.ThesisItem Open Access Management of stripe rust of wheat incited by Puccinia striiformisf.sp.tritici westend(CCSHAU, 2019) Bilal, Mohammad; Rajender SinghThe present study entitled, ―Management of stripe rust of wheat (Puccini striiformis f.sp.tritici westend‖ was conducted during the 2017-2018 at research farm, Department of Plant Pathology, Chaudhary Charan Singh Haryana Agricultural University, Hisar. In present study, symptoms of stripe rust appeared after 14 days with uredospores of Puccinia. striiformisf. sp. tritici(Pst). Maximum stripe rust severity was recorded in Yamunanagar followed by Ambala and least was observed in Fatehabad. The varieties HD3086, WH1105, WH1124 and WH1142 were free from stripe rust. Two commonly prevalent pathotypes were 46S119 and 110S119. Maximum terminal disease severity was recorded by WH 147 and HD2967 in seedling and tillering stage disease appearance followed by stem elongation and least disease severity was observed on milk development. Maximum yield loss was obtained by WH 147 and HD2967. WH1105. WH1124, WH 1142, WH 1237, WH 1238, WH 1241, WH 1242, WH 1244, WH 1247, WH 1248, WH 1239, WH 1246, WH 1250, WH 1251 genotypes were found resistant. Minimum disease severity observed on application of Nativo75 WG (trifloxystrobin25%WG+ tebuconazole50%), @ 300 gm/ha and Tilt 25EC (propiconazole25EC) 500ml/ha on appearance of disease at stem elongation stage and maximum yield gain was obtained. There was no residual toxicity of propiconazole25%EC (Tilt25%EC) sprayed @ 0.1% on tillering and stem elongation stage.ThesisItem Open Access Studies on epidemiology and management of yellow vein mosaic virus disease of okra (Abelmoschus esculentus L. Moench)(CCSHAU, 2019) Pankaj; Vinod KumarThe present study entitled, “Studies on epidemiology and management of yellow vein mosaic virus disease of okra (Abelmoschus esculentus L. Moench)” was conducted during the kharif 2018 at research farm, Department of Plant Pathology, Chaudhary Charan Singh Haryana Agricultural University, Hisar. Okra yellow vein mosaic virus disease is a devastating viral disease which has a wide occurrence and causes considerable yield losses quantitatively and qualitatively. The viral disease can appear at all stages of crop growth depending upon favorable conditions. Environmental factors play important role in sustaining whitefly population and disease development. It was observed that whitefly population builds up started in month of July and reached to maximum in first week of October. The whitefly population and percent disease index (PDI) was observed maximum in first week of October due to prevailing favourable weather conditions, i.e. maximum temperature 34±2 °C minimum temperature 23±2 °C, morning relative humidity more than 90 per, wind speed about 3 km/h, more sunshine hours and no rainfall. The average whitefly population was maximum on Pusa Sawani (6.54) followed by Pusa A4 (5.27) and Varsha Uphar (3.78). All three cultivars had positive significant correlation with bright sunshine hours while morning relative humidity and maximum temperature was recorded positively non-significant. However, evening relative humidity, wind speed and rainfall was negatively (non-significant) correlated, while minimum temperature was recorded negatively significant with whitefly population. The maximum and minimum temperature, evening relative humidity, wind speed, and total rainfall were negatively correlated with OYVMV disease where morning relative humidity and bright sunshine hours were found positively correlated. The highly significant positive correlation was found between whitefly population and disease severity. The terminal PDI was observed highest in Pusa Sawani (73.00 %) cultivar and lowest in Varsha Uphar (18.75 %). The maximum PDI and disease incidence was observed in 14 July sown crop with wider row to row spacing while minimum in 15 June sown crop with closely spaced. For the management of OYVMV disease chemical, botanicals and resistance inducers were used. Amongst all foliar spray of malathion 50 EC @ 2ml/litre of water + sarpagandha leaf extract @10% was found most effective in managing the disease with PDI (15.55 %). Amongst the resistance inducers seed priming with salicylic acid @ 150 mg/litre and spray the same also found effective against OYVMV with PDI (28.89 %).ThesisItem Open Access Management of banded leaf and sheath blight caused by Rhizoctonia solani f.sp. sasakii of maize(CCSHAU, 2019) Yadav, Lokesh; Ashwani KumarThe present study entitled “Management of banded leaf and sheath blight caused by Rhizoctonia solani f.sp. sasakii of maize” was conducted during Kharif season of 2018 at CCSHAU, Regional Research Station Uchani, Karnal. All the laboratory work was carried out in the Department of Plant Pathology, College of Agriculture, Chaudhary Charan Singh Haryana Agricultural University, Hisar. The experiments were laid out in RBD in field and CRD in lab conditions. Maize (Zea mays L.) is the most important cereal crops in the world agricultural economy with high yield potential. The disease banded leaf and sheath blight of maize caused by Rhizoctonia solani f.sp. sasakii is considered as the main limiting factor for reduced production. The characteristic symptoms of disease appears on leaves and sheaths on 40-45 days old plants. Symptoms on leaf sheath were water-soaked, round in shape and straw-coloured whereas, on leaves irregular, water soaked, round to stretched lesions having diameter 1-3 mm were observed. In case of severe infection, the pathogen manifests itself on the cobs and results in no grain formation. It was observed that the growth of pathogen was white to creamy whitish on PDA after 24 hours of inoculation. The sclerotia were round, semi spherical to irregular in shape and were scattered in the Petri plates. To evaluate the efficacy of three different aqueous compost extracts were used under in vitro conditions for the per cent mycelial growth inhibition. It was found that the compost extracts of cow desi inhibited mycelial growth up to 22.22 per cent at 20 per cent concentration. It was revealed that seed treatment with carbendazim @ 2g/kg of seed was found most effective in controlling disease upto 54.25 per cent. Seed treatment with Trichoderma harzianum @ 0.2% and soil application with P. fluorescens @ 0.2% was most effective in controlling the disease upto 66.89 per cent and highest yield (57.68 q/ha) was also recorded from this treatment. Out of five fungicides evaluated, foliar sprays of azoxystrobin (0.1%) twice at 30 and 45 DAS was found most effective in reducing the disease upto 73.54 per cent and increasing grain yield upto 77.30 per cent. Out of twenty seven maize hybrids and sixty inbred lines of maize screened against banded leaf and sheath blight under field condition, only seven hybrids and two inbred lines showed resistant reaction against banded leaf and sheath blight disease of maize.ThesisItem Open Access Epidemiology and management of stem gall of coriander (Coriandrum sativum L.) caused by Protomyces macrosporus Unger(CCSHAU, 2019) Vijaykumar S; Kushal RajThe present investigation entitled "Epidemiology and management of stem gall of coriander (Coriandrum sativum L.) caused by Protomyces macrosporus Unger" was conducted in the Department of Plant Pathology, CCS HAU Hisar laboratories and research farm during year 2018-2019. Typical symptoms in infected plants were observed on all above ground parts of coriander. Symptoms appeared in the form of tumour-like swellings (galls) on stem and fruits. These tumours were smooth initially but later ruptured and turned to be rough. The pathogen Protomyces macrosporus was isolated as pure culture on PDA medium enriched with yeast extract (PDA+YE). The colony colour was creamy white and chlamydopsores were yellowish brown and spherical in shape with three distinct layers & diameter 40-80 μm. The pathogenicity was confirmed as the symptoms developed after 63 and 75 days of incubation with the application of two different source of inoculum. Epidemiology of pathogen was studied under field condition by sowing the crop at five different dates and disease severity on tagged plants and over all incidence was maximum in December 1st week sown crop with average temperature of 28.9°C and 80 per cent of relative humidity. Out of five fungicides evaluated against Protomyces macrosporus, Ridomil MZ and Blitox 50 WP were found most effective and significantly superior among the treatments with 100 per cent growth inhibition of the pathogen at 0.2 per cent concentration under in vitro conditions. Out of five plant extracts evaluated under in vitro conditions against the pathogen, datura extract at 10 per cent was found most effective in mycelial growth inhibition. Among three bioagents evaluated under in vitro conditions against the pathogen, Pseduomanas fluorescens was found most effective to inhibit the growth of pathogen. Seed treatment and foliar spray of Ridomil MZ and Blitox 50 WP at 0.2 per cent was found most effective to manage the disease severity and incidence, resulting in comparative higher yield under field conditions. Amongst 28 genotypes screened against stem gall of coriander, one genotype viz., ICS-4 was found resistant, eleven genotypes as moderately resistant and sixteen genotypes were moderately susceptible.ThesisItem Open Access Epidemiology and management of early blight of tomato incited by Alternaria solani(CCSHAU, 2018) Ravinder; Narender SinghEarly blight disease caused by Alternaria solani is one of the most important fungal diseases of tomato (Lycopersicum esculentum). Keeping in view the enormous loss caused by the pathogen, the present investigation was carried out to study the early blight disease progression in relation to weather parameters and its management by using the plant extracts, bio-agents and fungicides under both in vitro and field conditions during Rabi season, 2017-18 at CCS HAU, Hisar. Survey was conducted in eight villages of two districts (Bhiwani and Hisar) in Haryana and minimum per cent disease intensity was recorded in village Saral of Bhiwani district. The epidemiological study revealed that early sown crop was least affected by the early blight disease compared to the timely and late sown crop. The per cent disease intensity increased with the increased age of the plant. . Mancozeb (75% WP) and hexaconazole and zineb (72% WP) were found to be most effective among the fungicides under in vitro condition. Two plants extract (neem leaf extract and combination of garlic clove and green chili extract) and a bio-agent (Trichoderma viride) was also found effective in inhibiting the mycelial growth of A. solani among the plant extracts and bioagents, respectively. In field conditions, mancozeb 75% WP was found to be most effective in reducing the early blight disease intensity and increasing the fruits yield of tomato crop.ThesisItem Open Access Studies on some epidemiological and biochemical resistance parameters for powdery mildew disease in Indian mustard caused by Erysiphe cruciferarum Opiz. ex. Junell(CCSHAU, 2018) Sunil H. S.; Rathi, A.S.Powdery mildew disease caused by Erysiphe cruciferarum is an important disease of Indian mustard (Brassica juncea) particularly in northern and north eastern states of India. Keeping in view the importance of disease present study has been carried out on epidemiology in relation to weather, sources of resistance and biochemical changes due to powdery mildew infection under in field conditions during rabi 2017-18 at CCS HAU, Hisar. Powdery mildew disease first appeared in third week of February on all the three varieties (RH 0749, RH 0406 and RH 9801) sown on 16th October and delayed by a week i.e. in the 1st week of March in all the varieties sown on 26th October and 8th November, when the temperature ranged between 27.3-29.4°C (maximum) and 9.4-13.5°C (minimum), while relative humidity (RH) range was 91.7-94.0 (morning) and 37-49% (evening). Wind speed during this period ranged from 1.1-2.7 km/ha, while sunshine range was 5.4-7.7 hours. Maximum disease progression in last two dates of sowing was observed from March 4th to March 13th, when the temperature ranged between 28.2-28.7°C (maximum) and 10.7-12.3°C (minimum), while RH ranged between 81.0-91.3% (morning) and 35.7-48.3% (evening). Wind speed and sunshine hour during this period was 2.2-2.5 km/hour and 4.9-7.7 hours, respectively. Temperature (maximum and minimum), wind speed and sunshine were positively correlated with per cent disease intensity, while relative humidity (morning and evening) and average vapour pressure (morning and evening) were negatively correlated with the per cent disease intensity and AUDPC increased with delay in time of sowing. The apparent rate of infection was high during initial stages of disease development, while it was negligible from 2nd week of March in all the varieties. Varieties RH 0749 showed less disease intensity followed by RH 0401 and RH 9801. Out of 148 genotypes screened under the artificial inoculation conditions, 5 genotypes (HNS 0901, Bio 902, BioYSR, DRMRIJ 12-48 and JM-6011) were resistant and 21 genotypes were moderately resistant, 55 genotypes were moderately susceptible and 31 genotypes were susceptible, while rest of the genotypes were highly susceptible. Slightly higher amount of chlorophyll ‘a’, chlorophyll ‘b’ and total chlorophyll and total sugars was found in healthy leaves of resistant (DRMRIJ 12-48) as compared to susceptible (RH 30) genotype. After infection amount of chlorophyll (a, b and total) and total sugars reduced gradually in both genotypes, but reduction was more in susceptible genotype particularly under inoculated conditions. The amount of phenols (total and OD) and the activities of POX and PPO were increased at higher rate in resistant as compared to susceptible one at the time of disease initiation. At the time of maximum disease intensity i.e. at 120 DAS, the activities of enzymes were more in resistant genotype under inoculated condition in comparison to un-inoculated condition.ThesisItem Open Access Studies on variability and management of Verticillium fungicola causing dry bubble disease in Agaricus bisporus(CCSHAU, 2018) Jagdeep Singh; Surjeet SinghVerticillium fungicola is a serious pathogen causing dry bubble disease in button mushroom (Agaricus bisporus). Present investigations were carried out on both host and pathogen by covering aspects of variability in pathogen, host-pathogen interaction and management of the disease. The isolates of V. fungicola were collected from different mushroom farms of Haryana state, coded as MHS (Hisar), BFT (Fatehabad), NJN (Jind), RHT (Rohtak), TPN (Panipat), BSN (Sonipat), FDB (Fridabad) and SKK (Kurukshetra) and pathogenicity was proved on A. bisporus. All isolates showed morphological, physiological and pathological variations. The isolates BSN, TPN, FDB, SKK and RHT are fall in the fast growing category having radial growth of 44.66, 43.86, 43.33, 42.16 and 41.50 mm, respectively and whitish colonies with dark yellow underside on PDA at pH 6.5 and 25±1ºC temperature after 12 days of incubation. Similarly, during screening of the isolates, only BSN, TPN, FDB, SKK and RHT showed disease symptoms on fruiting bodies of all the strains of A. bisporous included in the study and other mushroom spp. i.e. A. bitorquis, Pleurotus sajor-caju and P. florida, except A. bisporus strain U-3 and Calocybe indica where no disease appeared. During interaction between A. bisporus and the virulent isolate (BSN) of V. fungicola, both are easily distinguishable at interaction regions on the basis of hyphal width i.e. ranged from 4.0 to 6.5 μm and 1.5 to 4.5 μm, respectively. Pathogen grows inter- as well as intra-cellularly on host hyphae, thereby causing coiling and lysis of host mycelia. During enzymatic bioassay in dual culture, the mycopathogen showed production of different hydrolytic enzymes i.e. amylase, cellulase and chitinase but not lipase and pectinase by formation of clear zonation on substrate. In in vitro studies three bacterial isolates i.e. BI, BII and BIII were isolated from casing soil for their antagonism against V. fungicola. The maximum growth inhibition of V. fungicola was 78.64% with BII isolate followed by 63.94% and 61.10% in BI and BIII, respectively after 12 days of incubation. The efficacy of neem products i.e. neem seed kernel extract, neem oil and neem leaf extracts, at three concentrations (2.5, 5.0 and 7.5 μl/ml) were determined against V. fungicola. The radial growth inhibition recorded was 50.02% at 7.5 μl/ml in case of neem seed kernel extract, followed by 40.99% and 34.94% in neem leaf extract and neem oil, respectively. Salicylic acid and jasmonic acid with different concentrations (0.1, 0.2 and 0.4 mM) were used and growth inhibition was 44.96% recorded at concentration 0.4 mM with salicylic acid, followed by jasmonic acid (23.42%) when evaluated against V. fungicola. In In vivo both salicylic acid and jasmonic acid resulted in reduction of lesions size on A. bisporus to the extent of 69.69% and 39.93%, respectively at concentration of 0.4mM. Similarly, the number of lesions reduction was 81.96% (salicylic acid) and 54.64% (jasmonic acid) at same concentration. The integrated managements of dry bubble disease caused by V. fungicola during the cultivation of A. bisporus in cropping period of 2016 and 2017 showed that all the treatments individually as well in combinations affected all yield parameters at all stages of growth of A. bisporus. However, the integrated disease management reported when treatments combination (bacterial isolate BII, neem seed kernel extract and salicylic acid) were applied at both spawning and casing time resulted in maximizing yield (24.75%) and minimizing disease incidence (4.31%) in cropping duration of 37 days.
