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Open Access
Studies on variability and management of Verticillium fungicola causing dry bubble disease in Agaricus bisporus
(CCSHAU, 2018) Jagdeep Singh; Surjeet Singh
Verticillium 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.
Open Access
Ecofriendly management of Rhizoctonia solani causing black scurf of potato
(CCSHAU, 2018) Manoj Kumar; Anil Kumar
In recent years, among different fungal diseases of potato, black scurf disease has emerged as a serious problem caused by Rhizoctonia solani in most of the potato growing areas of India with 2-70 per cent disease incidence leading to considerable yield losses up to 50 per cent. Due to difficulties in disease management and lack of stable genetic resistance in potato cultivars against R. solani, different ecofriendly management aspects including fungicides, bioagents, organic amendments and cultural practices as well as systemic acquired resistance (SAR) activators in vitro and in vivo conditions were undertaken for effective management of the disease in present investigation. The in vitro and in vivo management study involving fungicides, bioagents or organic amendments indicated that among fungicides, Monceren, MEMC were most effective, whereas, in bioagents Trichoderma harzianum produced better results followed by T. viride. Among the organic amendments, vermicompost was found most effective followed by neem cake against black scurf of potato. Besides chemicals and biological control, cultural practices have also been used to minimize the inoculums level from the soil and tubers. Black scurf incidence is significantly influenced by irrigation level and depth of planting. As the irrigation frequency and depth of planting were increased, disease incidence and severity of potato black scurf was also increased. The delayed date of planting also affects disease incidence and severity. It was observed that soil moisture content is directly proportional to increased irrigation frequency and depth of planting. Black scurf disease incidence and severity showed a linear and negative correlation with soil temperature, whereas, a linear and positive correlation with soil moisture. Biochemical analysis indicated roles of systemic acquired resistance (SAR) inducing chemicals viz., salicylic acid, jasmonic acid, β-aminobutyric acid, γ-aminobutyric acid and hydrogen peroxide against black scurf of potato caused by R. solani. SAR activators play an important role in biochemical composition viz., total phenol, peroxidase, polyphenol oxidase and phenylalanine ammonia lyase and enhanced defense mechanism in potato crop for black scurf. It was also inferred from the study that all the biochemical constituents initially increased up to peak concentration after that, it decreased. Tubers dipping of SAR activators were found more effective to control the disease by reducing disease incidence as compared to foliar spray.
Open Access
Variability and management of rhizoctonia solani kuhn, the incitant of sheath blight of rice
(CCSHAU, 2018) Pankaj Kumar; Anil Kumar
Present investigation entitled ―Variability and management of Rhizoctonia solani Kuhn, the incitant of sheath blight of rice‖ revealed that Rhizoctonia solani isolates showed a considerably variation in cultural and morphological characters and pathogenic behaviour. There was no correlation of geographic location and source of isolation of the pathogen of the fungal isolates with the cultural, morphological and pathogenic clustering. ITS species specific primers indicated the intraspecific variation within R. solani isolates. R. solani was most dominant species associated with sheath blight complex in Haryana. However, R. oryzae was also found associated to the extent of 1.5%. Out of 307 rice genotypes evaluated, only one rice genotype i.e. RMS-BL-6 showed consistent resistant reaction against sheath blight during both years, indicating that resistance against sheath blight is rarely found. Among botanicals aqueous extracts of garlic cloves and turmeric powder proved effective in reducing disease severity while among fungicides azoxystrobin 18.2% + difenoconazole 11.4% SC and thifluzamide 23.9% SC proved to be the best treatments. None of the fungicide persisted till the harvest of crop i.e. 44 days after their last spray application in rice grains, cropped soil, paddy straw and husk indicating that these fungicides are safe for human, animal and environmental point of view. Among the combination of botanicals and fungicides, application of azoxystrobin 18.2% + difenoconazole 11.4% SC (1 ml/l) followed by extract of garlic cloves or turmeric powder at 50 g/l proved most effective but found out inferior to azoxystrobin 18.2% + difenoconazole 11.4% SC and thifluzamide 23.9% SC. All the herbicides inhibited the mycelial growth, sclerotial formation and sclerotial germination in vitro. Under field conditions pretilachlor 50 EC and pendimethalin 30 EC reduced the sheath blight severity alongwith significant increase in grain yield of paddy.
Open Access
Studies on maydis leaf blight of maize caused by Bipolaris maydis (Nisikado and Miyake) Shoemaker
(CCSHAU, 2017) Manjeet Singh; Mehra, Rakesh
The maydis leaf blight disease of maize (Zea mays L.) caused by Bipolaris maydis has emerged as a serious problem in most of the maize growing places of India, leading to considerable yield losses in maize. The present investigation on epidemiology, management and variability (cultural, morphological, pathogenic and genetic) were carried out with twenty six isolates of B. maydis collected from maize growing areas of the country. Out of four dates of sowing of maize, the highest disease intensity, AUDPC and AIR was observed in Ist date of sowing (15 June) followed by IInd date of sowing (27 June), IIIrd date of sowing (10 July) and least disease intensity in IVth date of sowing (25 July) on all the five hybrid and inbreds. Maximum temperature range of (32.2-34.20C), minimum (23.7-25.30C) with relative humidity morning (88-93%) and evening (59.5 -76.7 %), rainfall (0.0-7.4 mm), evaporation (2.6-3.9 mm), sun shine (4.8- 9.7 h/day) and wind speed (2-3.4 km/h) were most congenial for disease progression during Kharif 2015 and 2016. Temporal increase was maximum in 33th to 34th and 36th to 37th metrological weeks in all the five hybrid/inbreds during both the years. Disease progression, AUDPC and AIR value were more in Kharif 2016 as compare to Kharif 2015 however, among the five hybrid/inbreds highest in HKI-PC4B and minimum on HM-4 in all four date of sowing during Kharif 2015 and 2016. Relative humidity, rainfall and temperature showed important role in disease progression. Regression equations on relationship between disease development and weather parameter have been developed for five maize hybrid/ inbreds. Out of one hundred and two inbred lines and one hundred and four hybrids of maize screened against maydis leaf blight under artificial inoculated conditions, six inbreds and eighteen hybrids found resistant respectively. Out of eight fungicides evaluated against B. maydis, propiconazolee and tebuconazole provided maximum per cent growth inhibition, disease control and also increase yield. Among twelve plant extracts evaluated against B. maydis, garlic clove and neem leaves extracts at 20 per cent concentration were found most effective both under in vitro and in vivo conditions. Bio agents, Trichoderma harzianum and T. viride found most effective under in vitro, screen house and field conditions. For integrated disease management, combination of propiconazolee (0.1%) + garlic (20%) + T. harzianum and tebuconazole (0.1%) + neem (20%) + T. viride were most effective and also increase the yield. All the twenty six isolates identified, belonged to B. maydis based on their morpho-cultural characteristics. Among the isolates, BM-22 and BM-23 from Gogunda and Fateh Nagar recorded maximum colony diameter. The growth pattern varies among twenty six isolates of B. maydis showed rough or smooth texture, regular or irregular margin with or without zonation, mycelium of isolates was either appressed or raised type. The colors of the colony were highly variable and size of conidia ranged 28.38-77.40 × 10.32-23.22μm. The conidia size of isolate BM-20 was largest (63.98 × 15.74 μm) with highest average number of' septa (5.9). In pathogenic variations, the isolate BM-16 from Karnal was most virulent and aggressive in comparison to other twenty five isolates of B. maydis, as it showed shortest mean latent period (39.6 h) and maximum mean disease score (4.1) across ten maize genotypes. Molecular variability among the twenty-six isolates of B. maydis has been examined through RAPD and ITS analysis, isolate BM-3 isolate to remain alone in the dendrogram, whereas maximum closeness (97%) was observed between BM-24 and BM-25 isolates.
Open Access
Epidemiology, morpho-pathogenic variability and management of white rust of Indian mustard incited by Albugo candida (Pers. ex. Lev)
(CCSHAU, 2017) K. Basavaraj; Rathi, A.S.
White rust disease caused by Albugo candida is an important disease of Indian mustard (Brassica juncea.) in India particularly in northern and North eastern states. Keeping in view the enormous loss caused by the pathogen, the present study has been devised on epidemiology, variability and management studies under field and screen house conditions during rabi 2014-15 and 2015-16 at CCS HAU, Hisar. In epidemiological studies the two Indian mustard varieties namely RH 30 and RH 0749 were sown in different dates to observe the influence of weather factors on onset and progression of the disease. The white rust disease appeared early in the season i.e. last week of December in first three dates of sowings while a week delayed in late sown crops. Initially, after the onset of the disease, disease was slow and non-significant; however it gained momentum and showed significant difference among the observations in all dates of sowing from second week of January. The highest disease intensity was observed during the third week of January. The disease was rapid and more severe on variety RH 30 as compared to RH 0749. The severity was observed to be more with the delayed sowing. Investigation on selective criteria for Indian mustard genotypes for white rust intensity and yield revealed that, existence of great variability among the all the genotypes for white rust intensity and yield characters under all the three environments such as natural, artificially pathogen inoculated and pathogen controlled environment. The disease severity was more in environment where the zoosporangial suspension was artificially inoculated to create a disease pressure compared to naturally infected and controlled environments. In general the genotypes RH 9801, RH 9304 recorded the minimum disease severity while the genotypes RB 50, RH 0725, RH 0673 and RH 1134 were found to be highly susceptible. The genotypes RH 8812, RH 9304, RH 0749 performed better with uniform superiority under both no disease stress and disease stress conditions. So such genotypes can be encouraged to use in breeding programme. The variability among the Albugo candida isolates collected from the six different locations of Northern India and also from the different Brassica species from the Hisar showed considerable variation in morphology and pathological reactions on set of Brassica host species. The isolates showed greater variation in size and germination percentage of sporangia; however very less variability was observed for the shape of sporangia in which all the isolates exhibited spherical to globose. Based on the pathogenic reaction on set of Brassica species, Hisar, Bawal, Bharatpur and Ludhiana were seems to be similar. The Pantnagar and Delhi isolates seems to be entirely distinct isolates from Northern India. The results of cross inoculation studies indicated about existence of variability in the isolates and five groups were identified among the isolates. Among the different fungicides tested for their efficacy against white rust, spray of mancozeb 75 WP 45 DAS @0.2% followed by azoxystrobin 23% SC @ 0.05% at 60 DAS gave maximum control of the disease at both leaf and staghead phase with higher seed yield.
Open Access
Studies on blue mould rot of aonla (Emblica officinalis Goerth.) caused by Penicillium islandicum (Sopp.)
(CCSHAU, 2017) Saini, Anil Kumar; Sharma, Sushil Kumar
Aonla or Indian gooseberry (Emblica officinalis Goerth. Syn. Phyllanthus emblica L.) is one of the most important indigenous fruit of Indian sub continent. The blue mould rot caused by Penicillium islandicum adversly affects the fruit quality, quantity and ultimately reduces the marketable value. In India, nearly 20 – 35 per cent of perishables are lost due to post harvest diseases. Keeping the above facts in view, the present study was undertaken to survey orchards and terminal markets of Hisar and Rewari districts of Haryana, screen different varieties, study the various biochemical and epidemiological factors, and to develop suitable strategies through use of fungicides, chemicals, bioagents and botanicals for management of blue mould rot of aonla. Fortnightly survey carried out from November to January, 2015-16 revealed that blue mould rot incidence was higher in terminal markets of Hisar and Rewari Distt as compared to orchards in the respective locations. Varietal screening revealed that least blue mould rot intensity was found in Desi (2.67%) and Kanchan (3.33%) while maximum rot was observed in Chakaiya (50.00%) and Banarsi (49.33%) aonla varieties. In biochemical studies, it was observed that total soluble solids, total phenol, peroxidase and polyphenol oxidase activity increased in diseased fruits in comparison to healthy fruits, while ascorbic acid and acidity decreased over time in both healthy and diseased fruits. The epidemiological studies revealed that the injury of the aonla fruits was found to be a pre-requisite for infection. A minimum number of 50 spores/ml was necessary for disease initiation whereas maximum blue mould rot incidence was noted at 30ºC and 100 per cent relative humidity. Relative efficacy of five chemicals against blue mould rot tested in vitro and in vivo at 0.2, 0.5 and 1.0 M concentrations revealed that boric acid at 1.0 M concentration was the best to inhibit growth in pre- and post-inoculation after five and ten days of inoculation in both test conditions. In vitro and in vivo bio-efficacy studies involving ten plant extracts at 5, 10 and 20 per cent concentration for the management of blue mould rot indicated that treatment with neem leaf extract exhibited maximum mycelial growth inhibition (91.88%) at concentration of 20 per cent resulting in minimum pre- and post-treatment disease intensity of 7.31 and 8.13%, respectively, after five days of inoculation. Similar trend was also recorded in pre and post treatment after ten days of inoculation. Five bioagents were also studied for control of Penicillium islandicum by dual culture technique. Significantly highest per cent growth inhibition of P. islandicum was noted in Trichoderma harzianum (83.06%). T. harzianum with disease intensity 6.0 and 6.31 per cent was found most efficient antagonist in reducing the blue mould rot with disease intensity of 34.48 and 39.75% in pre- and post- inoculation, respectively, after five days of inoculation. Similar results were also obtained in pre- and post-treatment after ten days of inoculation.
Open Access
Studies on variability and anastomosis among the potato and rice isolates of Rhizoctonia solani Kuhn
(CCSHAU, 2017) Pandit, Gurav Narayan; Surjeet Singh
Sheath blight of Rice and black scurf of Potato caused by Rhizoctonia solani has emerged as a serious problem in majority of rice-potato growing states of India, leading to considerable yield loss in both rice and potato crops. The present investigation on variability, anastomosis and cross infectivity was carried out with eighteen rice and eleven potato isolates of R. solani collected from different locations. Based on the radial colony growth rate, all the isolates from both the crops were categorised in to three groups as slow, medium and fast growing. Majority of rice isolates showed medium colony growth rate, while, most of the potato isolates showed slower growth rate. Potato isolates had colony texture either appressed or raised, whereas rice isolates had either raised or fluffy growth. Colony colour in all isolates was varied from pale yellow to very pale brown, while whitish color was appeared in rice isolate R16. Sclerotia were found aggregated at the centre of the colony in potato isolates, whereas it was scattered on colony surface in rice isolates. Colour of sclerotia in all rice isolates was dark brown while it was brown in Kurukshetra (R4) and black in Dapoli isolate (R18), however, it was varied from light brown to dark brown in potato isolates. Moderate to profuse sclerotial intensity was observed in rice isolates whereas it was scanty to moderate in potato isolates. Among the rice isolates, mean maximum lesion length was recorded in isolate R14 (Ludhiana isolate) which indicated strong pathogenic nature towards all four cultivars while lowest mean lesion length was observed in isolate R12 (Gurdaspur isolate) which indicated the least virulent nature.. Among the potato isolates, isolate P5 was found highly pathogenic to all the four varieties while the isolate P10 was found least pathogenic. On the basis of mean lesion length in rice and per cent disease severity in potato, all isolates from both hosts were categorized into three virulent groups viz., VG-I (less virulent), VG-II (moderately virulent) and VG-III (more virulent). Majority of rice isolates were moderately virulent while most of the potato isolates were less virulent. The diversity analyses using ISSR primers, exhibited genetic relatedness among the eighteen rice isolates ranged from 61 to 82 per cent with an average similarity of 70 per cent. The ISSR analyses differentiated R. solani isolates into two major clusters at similarity coefficient of 0.61. The maximum genetic similarity (82%) among all rice isolates was observed between isolate R4 and R5 while minimum genetic similarity (66& 66.5%) with all other isolates was observed by isolates R11 and R2, respectively. Similarly, the genetic relatedness among eleven potato isolate ranged from 63 to 86 per cent with average similarity of 70 per cent. On the dendrogram, two main clusters viz., group A and group B, were produced at 0.63 similarity coefficient. The highest genetic similarity (86%) was observed between isolate P1 and P2, whereas minimum (69.5 %) with all other isolates was shown by isolate P9. The present investigation on cross-infectivity of rice and potato isolates indicated that none of the potato isolates were able to infect all four rice cultivars. On the other hand, rice isolates showed varied degree of virulence on three potato cultivar. In anastomosis interaction studies, majority of isolates showed compatibility with isolates with those belonged to same host with few exceptions. There was no compatible reaction occurred between rice and potato isolates indicated that both the isolates belong to different and distinct groups within same species.
Open Access
Integrated management of root rot of sesame (sesamum indicum L.) caused by macrophomina phaseolina (Tassi) goid
(CCSHAU, 2012) Bagaria, Pravin Kumar; Sharma, S. K.
Root-rot/charcoal-rot caused by Macrophomina phaseolina (Tassi) Goid. is one of the most important soil-borne disease in south-western parts of Haryana. Efforts were made to manage the disease with application of various methods. Root-rot incidence (seedling and maturity stage) and intensity (at maturity) was observed maximum in sandy soil and decreased with increase in clay content. The minimum disease incidence (pre-emergence, post-emergence damping-off, total seedling mortality) and intensity of charcoal rot was observed when optimum irrigation was applied at daily and after every 3rd day as compared when the pots were irrigated after every 7th and every 10th day. Carbendazim (50, 100, 200, 500 ppm), propiconazole (100, 200, 500 ppm) and hexaconazole (200, 500 ppm) exhibited complete (100%) mycelial growth inhibition of M. phaseolina in vitro conditions. The evaluation of 11 plant extracts at 5, 10, 20 and 40 per cent concentrations revealed that aqueous extract of garlic clove was proved to be most effective inhibitor of M. phaseolina in vitro. Onion bulb extract ranked as the next most effective in inhibiting mycelial growth of pathogen followed by extracts of bougainvillea, clerodendron and datura leaves. All antagonists viz., Trichoderma harzianum, T. viride and Pseudomonas fluorescens significantly reduced mycelial growth of M. phaseolina in vitro. However, T. viride and T. harzianum were proved the best with 70.27 and 67.5 per cent mycelial growth inhibition of M. phaseolina, respectively. Neem leaves (50 g/kg soil) was the most effective in reducing root-rot incidence/intensity under screenhouse conditions followed by clerodendron and bougainvillea leaves amendments. Neem seed powder and neem cake each @ 20 g/kg soil applied as organic amendment showed significant reduction in root-rot incidence/intensity as compared to control followed by jamun seed powder and castor cake. Dry seed dressing (1 g/kg seed) and soil application (5 g/kg soil) of bioagent, Trichoderma viride at various intervals produce lesser pre-emergence, post-emergence damping-off and charcoal rot incidence/intensity at maturity compared to control. Seed treatment as well as spray @ 100 ppm with either Isonicotinic acid (INA) or Salicylic acid (SA) was also found effective in reducing disease incidence/intensity under screenhouse conditions. Seed treatment with carbendazim (0.25%) showed the lowest disease incidence and intensity as compared to other treatments in the screenhouse and field conditions. In artificially inoculated conditions also (inoculated at pith region), seed treatment with carbendazim (0.25%) and soil application of neem cake (0.5 kg/m2) were the most effective. IDM (Integrated Disease Management) modules developed revealed that, soil application of neem cake (0.5 kg/m2) + Trichoderma viride (0.2 kg/m2) followed by seed treatment with carbendazim (0.25%) showed the lowest charcoal-rot incidence and intensity under field conditions.
Open Access
Studies on variability in sclerotinia sclerotiorum (Lib.) de bary causing stem rot in Indian mustard
(CCSHAU, 2013) Pawan Kumar; Rathi, A.S.
The stem rot disease caused by Sclerotinia sclerotiorum (Lib.) de Bary is the most serious fungal disease and causes maximum damage in Indian mustard (Brassica juncea). The present investigation on cultural, morphological, pathogenic and genetic variability was carried out with fourteen isolates of S. sclerotiorum collected from mustard growing areas of the country. All the isolates exhibited variations in their colony colour, type of growth and mycelial growth. HSR, FTH, SRS and DBW isolates showed dirty white colony colour, while rest of the isolates showed whitish colony colour. The variation in type of growth was also observed as BWL, MHR and BHR isolates showed fluffy and regular type of growth; ALW and JLN isolates showed fluffy but irregular growth, while rest of the isolates showed sparse and regular type of mycelial growth. All isolates showed significant differences in mycelial growth up to 72 hours of incubation, while non- gnificant differences were observed after 96 hours of incubation among the isolates. Significant variations were observed in number of sclerotia formation, size and pattern of sclerotia formation among the isolates after 10 days of incubation. Sufficient variability was also observed for number of apothecia per sclerotium. Isolates were also found to vary in their per cent disease intensity and significant host × pathogen interaction on different rapeseed-mustard genotypes under screen house and field conditions. HSR isolate was the most pathogenic with mean disease intensity of 53.1 and 60.8% under field and screen house conditions, respectively, while ALW was the least pathogenic isolate with disease intensity of less than 30% under both the conditions. Some isolates showed moderately resistant reaction on one genotype while, susceptible to highly susceptible reaction on rest of the genotypes. Some of the isolates could not be clearly differentiated pathogenically; however, they showed variable reactions on these genotypes. The variation was observed for stem breaking of different genotypes and formation of varying numbers of sclerotia after inoculation of S. sclerotiorum isolates under field conditions. The ITS (Internal transcribed spacer) analysis differentiated the fourteen isolates into two major clusters at similarity coefficient of 0.38. Cluster A was constituted by only one isolate ALW and Cluster B was a large group containing 13 isolates. Cluster B was further divided into two groups C and D which occupied nine and four isolates, respectively.

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