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ThesisItem Open Access RESPONSE OF COTTON GENOTYPES TO RELATION TO INDUCTION OF RESISTANCE AGAINST COTTON LEAF CURL DISEASE(PAU Ludhiana, 2013) Ritu Raj; P S, SekhonThe present study was conducted to check the response of different American cotton cultivars namely RS 921, LH 2076, PIL 8, Ankur 3028 BGII and a desi cotton variety LD 694 to Jasmonic acid (JA) and Salicylic acid (SA) in induction of resistance against cotton leaf curl disease (CLCuD). Different cotton cultivars were grown in pots under screen cages. At four to six leaf stage potted plants of different cotton cultivars were treated with different concentration of JA and SA i.e. 50 μM, 100 μM, 150 μM, 200 μM respectively and water sprayed plants of corresponding genotypes served as control. Quantification of proteins was done using spectrophotometric method from the leaf samples collected at 24, 48, 72, 96 hrs and a week after treatment with JA and SA. Application of JA and SA resulted in the induction of proteins 24 hrs after treatment. After a week interval at 150 μM concentration of JA maximum protein induction of 19.9 mg/g fr. wt., 16.7 mg/g fr. wt., 19.6 mg/g fr. wt., 15.0 mg/g fr. wt. and 14.4 mg/g fr. wt. was recorded in RS 921, LH 2076, PIL 8, Ankur 3028 BGII and LD 694 respectively whereas, in case of SA 200 μM concentration showed maximum protein induction i.e 10.6 mg/g fr. wt., 9.3 mg/g fr. wt., 9.9 mg/g fr. wt., 9.0 mg/g fr. wt., 8.1 mg/g fr. wt. in RS 921, LH 2076, PIL 8, Ankur 3028 BGII and LD 694 respectively. In the induction of proteins JA was found to be more effective than SA. Electrophoretic study of cotton cultivars treated with 150 μM of JA and 200 μM of SA revealed the induction of Pathogenesis-related (PR) proteins ranging from 15-45 kDa along with some other proteins as well. JA and SA applied at different concentrations affected the CLCuD incidence and severity when inoculated with viruliferous whiteflies (Bemesia tabaci) in screen cages. Lower disease incidence as well as disease index was observed with JA when applied @ 150 μM and SA @ 200 μM. Disease incidence was 37%, 30%, 30% and disease index was 48%, 40%, 40% in RS 921, LH 2076, Ankur 3028 BGII at 150 μM concentration of JA whereas, at 200 μM SA disease incidence was 48%, 36%, 34% and disease index was 57%, 50%, 50% respectively in above mentioned cultivars and in their respective control values for disease incidence and disease severity were quite high. So, JA at 150 μM and SA at 200 μM concentration was found to be most effective in lowering the disease. Latent carry over detection of symptomless plants treated with 150 μM of JA and 200 μM of SA through PCR amplification using DNA specific primers confirmed the presence of virus in all the tested cotton cultivars except LD 694 which signified that PR proteins does not eliminate virus. JA and SA application resulted in imparting tolerance with the induction of PR proteins but does not lead to complete resistance against the disease.ThesisItem Open Access RESPONSE OF COTTON GENOTYPES TO JASMONIC ACID IN RELATION TO INDUCTION OF RESISTANCE AGAINST COTTON LEAF CURL DISEASE(PAU Ludhiana, 2013) Raj, RituThe present study was conducted to check the response of different American cotton cultivars namely RS 921, LH 2076, PIL 8, Ankur 3028 BGII and a desi cotton variety LD 694 to Jasmonic acid (JA) and Salicylic acid (SA) in induction of resistance against cotton leaf curl disease (CLCuD). Different cotton cultivars were grown in pots under screen cages. At four to six leaf stage potted plants of different cotton cultivars were treated with different concentration of JA and SA i.e. 50 μM, 100 μM, 150 μM, 200 μM respectively and water sprayed plants of corresponding genotypes served as control. Quantification of proteins was done using spectrophotometric method from the leaf samples collected at 24, 48, 72, 96 hrs and a week after treatment with JA and SA. Application of JA and SA resulted in the induction of proteins 24 hrs after treatment. After a week interval at 150 μM concentration of JA maximum protein induction of 19.9 mg/g fr. wt., 16.7 mg/g fr. wt., 19.6 mg/g fr. wt., 15.0 mg/g fr. wt. and 14.4 mg/g fr. wt. was recorded in RS 921, LH 2076, PIL 8, Ankur 3028 BGII and LD 694 respectively whereas, in case of SA 200 μM concentration showed maximum protein induction i.e 10.6 mg/g fr. wt., 9.3 mg/g fr. wt., 9.9 mg/g fr. wt., 9.0 mg/g fr. wt., 8.1 mg/g fr. wt. in RS 921, LH 2076, PIL 8, Ankur 3028 BGII and LD 694 respectively. In the induction of proteins JA was found to be more effective than SA. Electrophoretic study of cotton cultivars treated with 150 μM of JA and 200 μM of SA revealed the induction of Pathogenesis-related (PR) proteins ranging from 15-45 kDa along with some other proteins as well. JA and SA applied at different concentrations affected the CLCuD incidence and severity when inoculated with viruliferous whiteflies (Bemesia tabaci) in screen cages. Lower disease incidence as well as disease index was observed with JA when applied @ 150 μM and SA @ 200 μM. Disease incidence was 37%, 30%, 30% and disease index was 48%, 40%, 40% in RS 921, LH 2076, Ankur 3028 BGII at 150 μM concentration of JA whereas, at 200 μM SA disease incidence was 48%, 36%, 34% and disease index was 57%, 50%, 50% respectively in above mentioned cultivars and in their respective control values for disease incidence and disease severity were quite high. So, JA at 150 μM and SA at 200 μM concentration was found to be most effective in lowering the disease. Latent carry over detection of symptomless plants treated with 150 μM of JA and 200 μM of SA through PCR amplification using DNA specific primers confirmed the presence of virus in all the tested cotton cultivars except LD 694 which signified that PR proteins does not eliminate virus. JA and SA application resulted in imparting tolerance with the induction of PR proteins but does not lead to complete resistance against the disease. Key words: Cotton, Cotton leaf curl disease (CLCuD), Jasmonic acid (JA), Salicylic acidThesisItem Restricted EVALUATION OF HOST PLANT RESISTANCE AND CHARACTERIZATION OF VIRUS(ES) CAUSING LEAF CURL DISEASE OF CHILLI (Capsicum annuum L.) IN PUNJAB(2013) Itoo, Asif AhmadChilli is known to be affected by forty five viruses, among them whitefly transmitted begomovirus of family Geminiviridae is considered to be the most destructive virus in terms of yield loss, causing leaf curl disease of chilli. In the present study, survey conducted in major chilli growing areas of Punjab showed that chilli leaf curl disease (ChiLCD) is prevalent in all the four districts surveyed with incidence ranging from 11 to 64%; maximum severity recorded in the months July-August in the districts Patiala and Sangrur. A total of 155 genotypes were screened under natural conditions and the identified resistant stocks were subjected to artificial inoculation. Symptom variability and mixed infections of begomovirus along with beta DNA satellite molecule was found in samples collected from field. Five begomoviruses viz., tomato leaf curl Jodyebpur virus (ToLCJV), tomato leaf curl Palampur (ToLCPV), tomato leaf curl New Delhi (ToLCNDV), tomato leaf curl Karnatka (ToLCKV) and papaya leaf crumple virus (PaLCrV) were found to be associated with ChiLCD in Punjab. Typical begomovirus symptoms were observed in artificially inoculated chilli lines after five weeks of inoculation. Three lines Surian 2010, Perennial and Japanese Loungi were promising against tomato leaf curl Joydebpur virus causing ChiLCD. Symptomatic and asymptomatic weeds showed presence of begomovirus when subjected to rolling circle amplification (RCA) + polymerase chain reaction (PCR), except Calatropis and Parthenium. Weeds are potential reservoir of tomato leaf curl Joydebpur virus (Abutilon and Ageratum), papaya leaf crumple virus (Cannabis), chilli leaf curl India virus (Ipomea and Amranthus) in Punjab.ThesisItem Open Access EPIDEMIOLOGY AND MANAGEMENT OF CURVULARIA LEAF SPOT OF GLADIOLUS CAUSED BY Curvularia trifolii f. sp. gladioli PARMELEE AND LUTTRELL(Punjab Agricultural University, Ludhiana, 2013) HANUMAN SINGHThe fungus C. trifolii f. sp. gladioli did not survive in pots containing sterilized soil during the off-season. Colonies of fungus C. trifolii f. sp. gladioli could not be observed from the artificially inoculated soil by dilution plate method. Among fifty varieties evaluated in the field, none was found to be immune. However, nine varieties namely SJN, Yellow Stone, Plum Tart, Jacksonville Gold, Pink Elegence, Delhi Pink, Shagun, Arka Kesar and Punjab Flame showed very good level of genetic resistance (disease reaction: ≤ 1.0) against the disease. The varieties, namely Jester, Chandani, Punjab Dawn, Gunjan, Shobha, Solist, Jessica, Aldebaran, Purple Flora, Overture and Alexander the Great were susceptible to the disease (disease reaction: >3.0-4.0). Out of the nine fungicides evaluated in pots, all the fungicides except for Bavistin (0.1%) reduced the severity of the disease significantly over the control. In field, the minimum disease severity was recorded in Antracol, Score and Tilt (1.33), followed by Dithane M-45, Kavach and Topas (1.67). Moderate disease severity was found in Cuman-L, Blitox and Bavistin sprayed plant (2.33). The yield data showed that the highest number of corms/plot was recorded in Score (58.00); followed by Tilt (57.67), Kavach (56.33), Antacol (55.33), Dithane M-45 (54.33), Cuman-L (54.00), Bavistin (53.00), Topas (52.67) and Blitox (50.00).ThesisItem Open Access EPIDEMIOLOGY AND MANAGEMENT OF BLAST DISEASE OF RICE(Punjab Agricultural University, Ludhiana, 2013) Singh, Sukhjinder; Mohan, ChanderDisease surveys revealed that disease incidence and severity of rice blast on nursery plants of Pusa basmati 1121 cultivar varied from 15.0 to 40.0 per cent and 10.0 to 35.0 per cent respectively at different locations of Punjab during 2011 crop season. The basmati rice fields which had high disease severity at seedling stage in the villages of Deep Singh Wala (22.0 %), Sarala Kalan and Avan Khalsa (15.0 %) were also visited during the adult plant stage and disease severity in all the above fields varied only from traces to 4.0 per cent. Disease survey at adult plant stage revealed that the highest disease incidence and severity was recorded from village Dhurial (Jalandhar) and Murthli (Gurdaspur) on cultivar Pusa basmati 1121 during 2011 and 2012 crop season. Maximum colony growth and sporulation of Pyricularia grisea was observed on Oat meal agar medium. A temperature of 250C was found to be highly suitable for colony growth and sporulation of the pathogen. Minimum colony growth and sporulation was observed at 15°C. Maximum colony growth and sporulation was observed at pH 6.0 followed by pH 7.0. Among different substrates tested maximum sporulation was observed on makra grass followed by carrot cubes. A minimum of 12 hours of leaf wetness was required for infection and symptoms appeared within 7 days of incubation period. Thereafter, the incubation period decreased while disease severity increased significantly with increase in leaf wetness from 12 to 48 hours. A positive correlation between leaf wetness duration and disease severity was observed. Maximum increase in lesion length was recorded at 250C followed by 200C. Increase in lesion length was minimum at 100C. Under in vitro conditions, tebuconazole proved most effective followed by azoxystrobin + difenconazole, propiconazole and difenconazole. Under pot house conditions tebuconazole was most effective in reducing the disease followed by propiconazole. Under field conditions, propiconazole followed by azoxystrobin + difenconazole and tricyclazole were most effective in reducing the disease and increasing the paddy yield.ThesisItem Restricted EPIDEMIOLOGY AND MANAGEMENT OF BLAST DISEASE OF RICE(Punjab Agricultural University, Ludhiana, 2013) Sukhjinder SinghDisease surveys revealed that disease incidence and severity of rice blast on nursery plants of Pusa basmati 1121 cultivar varied from 15.0 to 40.0 per cent and 10.0 to 35.0 per cent respectively at different locations of Punjab during 2011 crop season. The basmati rice fields which had high disease severity at seedling stage in the villages of Deep Singh Wala (22.0 %), Sarala Kalan and Avan Khalsa (15.0 %) were also visited during the adult plant stage and disease severity in all the above fields varied only from traces to 4.0 per cent. Disease survey at adult plant stage revealed that the highest disease incidence and severity was recorded from village Dhurial (Jalandhar) and Murthli (Gurdaspur) on cultivar Pusa basmati 1121 during 2011 and 2012 crop season. Maximum colony growth and sporulation of Pyricularia grisea was observed on Oat meal agar medium. A temperature of 250C was found to be highly suitable for colony growth and sporulation of the pathogen. Minimum colony growth and sporulation was observed at 15°C. Maximum colony growth and sporulation was observed at pH 6.0 followed by pH 7.0. Among different substrates tested maximum sporulation was observed on makra grass followed by carrot cubes. A minimum of 12 hours of leaf wetness was required for infection and symptoms appeared within 7 days of incubation period. Thereafter, the incubation period decreased while disease severity increased significantly with increase in leaf wetness from 12 to 48 hours. A positive correlation between leaf wetness duration and disease severity was observed. Maximum increase in lesion length was recorded at 250C followed by 200C. Increase in lesion length was minimum at 100C. Under in vitro conditions, tebuconazole proved most effective followed by azoxystrobin + difenconazole, propiconazole and difenconazole. Under pot house conditions tebuconazole was most effective in reducing the disease followed by propiconazole. Under field conditions, propiconazole followed by azoxystrobin + difenconazole and tricyclazole were most effective in reducing the disease and increasing the paddy yield.ThesisItem Open Access EPIDEMIOLOGY AND MANAGEMENT OF ALTERNARIA BLIGHT ON SEED CROP OF CARROT (Daucus carota)(Punjab Agricultural University, Ludhiana, 2013) Gurpreet SinghSurveys conducted during 2011 and 2012 revealed that Alternaria blight of carrot was observed at all the locations on all the cultivars. The disease incidence and severity was the highest on Local varieties as compared to PAU recommended varieties. The highest disease severity was observed on local variety grown at village Bohan in district Hoshiarpur, while the lowest disease severity on Selection-21 grown at PAU Ludhiana. Alternaria radicina and Alternaria dauci were found to be associated with the disease. Alternaria radicina has highest frequency than Alternaria dauci on carrot plants. Maximum colony growth and sporulation of Alternaria radicina was observed on carrot leaf agar followed by unmbel extract agar medium. Lowest colony growth and sporulation was observed on Czapek‟s Dox agar medium. Maximum colony growth and sporulation was observed at 25°C temperature followed by 30°C, whereas no colony growth was observed at 35°C. Most suitable range of temperature was found to be 25 to 30°C. Maximum colony growth of Alternaria radicina was observed at pH 6, whereas maximum sporulation was observed at pH 7. The disease severity increase with the increase in the age of plants and a positive correlation between plant age and disease severity was observed. Similarly disease severity increased with the increase in the leaf wetness period. A positive correlation was found between leaf wetness period and disease severity. A minimum 8 hr of leaf wetness was required to cause infection on carrot plants. All three non systemic fungicides proved less effective than systemic fungicides. Among systemic fungicides ED50 values for difenconazole and propiconazole was less than 1μg/ml, while for hexaconazole and azoxystrobin was 1.0 and 5.9 μg/ml respectively. Among non systemic fungicides ED50 value was 17 for chlorothalonil, whereas it was 44 μg/ml for mancozeb. Under field conditions all systemic fungicides proved highly effective in controlling the disease as compared to non systemic fungicides. Azoxystrobin + Difenconazole @ 0.1 per cent proved most effective in checking the disease than other tested fungicides.ThesisItem Open Access ROLE OF GUTTATION FLUID IN THE SPREAD OF BACTERIAL BLIGHT OF RICE(Punjab Agricultural University, Ludhiana, 2013) Raj kiranThe role of guttation fluid on bacterial blight development and spread was investigated at different crop growth stages and nitrogen doses applied. Maximum amount (4.0ml/plant) of guttation fluid was collected from the plants inoculated at tillering stage. Similarly highest population density of Xanthomonas oryzae pv. oryzae (5.28 log cfu/ml) was also recovered in guttation fluid collected from X. oryzae pv. oryzae inoculated plants at tillering stage as compared to seedling and booting stage of crop. High bacterial load in guttation fluid were also reflected in high bacterial blight severity (12.2 %) and spread (up to 60cm from inoculated plant) at tillering stage. However, the disease failed to develop when the plants were inoculated at booting stage. Morning rain simulation washed off the guttation fluid from the leaf surface which resulted in significantly lesser disease severity and spread on rain simulated plots as compared to non rain simulated plots. When rain was simulated at tillering stage in bacterial blight susceptible cv. Pusa 1121, there was increase in population densities of X. oryzae pv. oryzae in guttation fluid collected from no rain simulated (6.28 log cfu/ml) plants as compared to rain simulated (5.84 log cfu/ml) plants after 7 days of inoculation. Application of high nitrogen doses significantly increased the disease severity and spread. Increased amount of guttation fluid per plant was observed in plots supplied with enhanced nitrogen doses. This also resulted in increase in total bacterial load per plant. Highest mean log cfu of X. oryzae pv. oryzae was obtained from treatment where nitrogen was applied @ 72 kg urea/acre (6.02 log cfu/ml) while minimum (4.72 log cfu/ml) was recorded from the plots when no nitrogen was applied. Electron microscopic studies also revealed intense colonization by X. oryzae pv. oryzae in rice leaves supplied with high nitrogen dose. Disease spread was also observed to maximum in plots supplied with 72 kg urea/acre (up to 80 cm from inoculated plant). A Bio-PCR based laboratory protocol was developed and tested for the detection of latent presence of X. oryzae pv. oryzae in the guttation fluid collected from asymptomatic plants.ThesisItem Open Access GENETIC ANALYSIS FOR STRIPE RUST (Puccinia striiformis f. sp. tritici) RESISTANCE IN WHEAT(Punjab Agricultural University, 2013) Avi KumarFifty four (54) advanced lines of wheat were characterized for resistance against two pathotypes of stripe rust pathogen namely, 46S119 and 78S84 at seedling and adult plant stage. Six lines viz ; PBW 679, BWL 0813, BWL 1638, BWL 1639, BWL 1653 and BW 9129 were found to be resistant against both the pathotypes at seedling and adult plant stages. Nine lines viz ; PBW 550, PBW 682, BWL 0136, BWL 0827, BWL 0883, BWL 0892, BWL 0893, BWL 1622 and BW 9065 showed seedling resistance but were susceptible at adult plant stage against 46S119 pathotype, while against 78S84 pathotype seventeen lines namely, PBW 669, PBW 670, PBW 678, PBW 681, PBW 685, PBW 687, BWL 0883, BWL 0926, BWL 0927, BWL 0928, BWL 1619, BWL 1620, BWL 1628, BWL 1641, BWL 1642, HD 2733 and BW 9065 showed seedling resistance but at adult plant stage, these were found to be susceptible. Three lines namely, PBW 680, PBW 683 and BWL 0831 were susceptible at seedling stage but showed high adult plant resistance. Genetic basis of resistance in cultivars PBW 621 (DPW 621-50) and HD 2967 revealed that PBW 621 possesses one gene for resistance and HD 2967 contains two genes for resistance against 78S84 pathotype of stripe rust pathogen of wheat, which is depicted from F3 segregating population of the crosses (PBW 621 x WL 711, HD 2967 x WL 711 and PBW 621 x HD 2967).