Browsing by Author "Sindhu, S.S."
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ThesisItem Open Access Amelioration of salt stress in chickpea (Cicer arietinum L.) by inoculation of ACC deaminase-containing mesorhizobia and rhizobacteria(CCSHAU, 2013) Chaudhary, Deepika; Sindhu, S.S.Chickpea is a major legume crop grown in the semi-arid tropics of Asia and Africa. Salinity mainly affects plant growth by decreasing the availability of water to the roots due to osmotic effect of external salts. Salinity also influences other physiological processes such as seed germination, photosynthesis, respiration and metabolite accumulation. The use of plant growth-promoting rhizobacterial as inoculants have been reported to facilitate plant growth in saline lands. Fifty isolates of Mesorhizobium were obtained from the nodules of chickpea plant samples and fifty five isolates of rhizobacteria were obtained from the chickpea rhizosphere soil collected from different part of Haryana. Among the fifty Mesorhizobium isolates, only four isolates i.e., MHD2, MSD41, MHD12 and MHD14 showed growth up to 4% NaCl salt concentration whereas 28 isolates among the 55 rhizobacterial isolates showed growth up to 4% salt with different colony size. Two Mesorhizobium isolates i.e., MHD1 and MHD12 and six rhizobacterial isolates i.e., RSD17, RSD19, RSD23, RBD12, RHD2 and RHD18 showed significant growth on ACC supplemented plates as compared to ammonium sulphate incorporated medium plates. Inoculation of selected Mesorhizobium or rhizobacterial isolates on sterilized chickpea seeds in water agar plates showed that isolate KR48 showed maximum growth of seedling roots at 5 days whereas MBD26 showed maximum growth of shoot at 10 days. Isolates RHD18, RSD3, RBD12 and RSD23 showed significant stimulation of shoot growth even at 8 dS/m salt concentration as compared to uninoculated control. Coinoculation studies with ACC+ as well as ACC- isolates of Mesorhizobium and rhizobacterial isolates were made in chickpea cultivar HC-1 under chillum jar conditions containing sloger’s broth with salt (EC, 4dS/m) and without salt. Coinoculation of ACC+ Mesorhizobium isolate MBD26 with rhizobacterial isolate RHD18 produced maximum 59 nodules/plant and 50.6% increase in plant dry weight and in the presence of salt slight decrease in nodulation (49 nodules/plant) and gain in plant dry weight (39.3%) was observed in comparison to single inoculation of MBD26 at 50 days of plant growth. At 80 days of plant growth, coinoculation of both ACC- MBD20 and RBD19 formed 41 nodules/plant and only 7.8% increase in shoot dry weight was observed as compared to the single inoculation. Coinoculation of Mesorhizobium isolate MBD26 with rhizobacterial isolate RHD18 caused increase in nodule number (78 nodules/plant), nodule weight (413 mg/plant) and shoot dry weight (61.7% increase). Whereas in the presence of salt, same treatment formed 53 nodules per plant and caused only 53.2% increase in plant biomass in comparison to single Mesorhizobium inoculated plants at 80 days of plant growth.ThesisItem Open Access Aminolevulinic acid production by rhizobacteria: its role in salt tolerance and weed control in mustard [Brassica juncea (L.)](CCSHAU, 2016) Phour, Manisha; Sindhu, S.S.In the present study, one hundred sixty two bacterial isolates were obtained from mustard rhizosphere soil. These bacterial isolates along with two reference strains were screened for production of amino-levulinic acid (ALA). Significant ALA production (> 15 µg/ml) was observed in bacterial isolates HMM21, HMM22, HMM80, HMM86, HMM92, HMM97, HMM115, JMM11, JMM15 and JMM35. Other eighty four isolates produced ALA ranging from 10.1 to 15.0 µg/ml. Ninety four bacterial isolates were selected on the basis of variation in ALA production ability and tested for salt tolerance at 1, 2, 4, 6 and 8% NaCl concentration. At 1% NaCl conc., ninety three bacterial isolates showed larger colony size ranging from 10.1-20 mm. At 4% NaCl concentration, only fifty six rhizobacterial isolates showed large colony size whereas only seventeen bacterial isolates showed large colony size at 6% NaCl concentration. Eleven bacterial isolates HMM6, HMM13, HMM34, HMM39, HMM44, HMM50, HMM65, HMM88, HMM92, JMM15 and JMM42 showed larger colony size (ranging from 10.1-20 mm) even at 8% NaCl concentration. Forty five rhizobacterial isolates were selected to study their inoculation effect on mustard seedlings on 0.8% water agar plates having different salt concentrations i.e., 0, 4, 6 and 8 dS/m. Eight rhizobacterial isolates HMM39, HMM44, HMM55, HMM57, HMM65, HMM73, HMM92 and JMM15 showed significant growth stimulation effect on seedling growth of mustard in comparison to uninoculated control at both 5 and 10 days of growth at different salt concentrations. These bacterial isolates also exhibited stimulation of root and shoot at 16 and 20 dS/m salt concentration except isolate HMM92, at both 5 and 10 days. Eleven isolates i.e., HMM21, HMM57, HMM76, HMM83, HMM92, HMM109, HMM116, JMM4, JMM24, JMM35 and CPS67 showed maximum retardation effect on seedling growth of Lathyrus aphaca. Based on their effect on growth of mustard and weed seedlings, eighteen rhizobacterial isolates were studied for other plant growth promoting activities. ACC utilization was observed only in eight bacterial isolates i.e., HMM8, HMM39, HMM57, HMM73, HMM92, JMM15, JMM19 and JMM24. HCN production was observed in two bacterial isolates JMM19 and JMM24. The IAA production was observed in all the isolates and it varied from 0.27 to 12.24 µg/ml. Thirteen bacterial isolates were found to possess phosphorus solubilizing activity. Three bacterial isolates i.e., HMM22, JMM15 and JMM19 were found to possess potassium-solubilizing index (K -SI) more than 5.0. Four bacterial isolates i.e., HMM92, JMM11, JMM19 and JMM24 were found to inhibit growth of two fungi Alternaria brassicae and Fusarium oxysporum. Eleven rhizobacterial isolates/strains were tested for their effect on growth of mustard and weed under pot house conditions. Rhizobacterial isolates/strain i.e., HMM76, HMM92, JMM24, JMM35 and SYB101 were found to stimulate growth of mustard along with inhibition of the growth of Lathyrus aphaca. At 75 days after sowing, inoculation of two bacterial isolates HMM92 and JMM24 showed 54 to 191% increase in RDW and SDW of mustard whereas they caused 36 to 92% decrease in RDW and SDW of Lathyrus aphaca. Rhizobacterial isolates i.e., HMM39, HMM57 and JMM15 stimulated growth of mustard 0, 8 and 12 dS/m salinity under pot house conditions. At 80 days after sowing, inoculation of three bacterial isolates HMM39, HMM57 and JMM15 caused 139 to 291% increase in RDW and SDW of mustard even at 12 dS/m. Based on the comparative analysis of various morphological and biochemical characteristics, the bacteria were found to belong to the genera of Aeromonas, Bacillus, Bacteroides, Brochothrix, Chrysemonas, Clostridium, Fusobacterium, Flavimonas, Hafnia, Mitsuokella, Meniscus, Photobacterium, Proteus, Pseudomonas and Serratia. Bacterial isolates JMM15, JMM24, HMM57 and HMM92 were identified as Pseudomonas azotoformans, Bacillus flexus, P. argentinensis and P. entomophila, respectively by the 16S rRNA sequence analysis. These bacterial isolate could be further exploited for growth improvement of mustard under field conditions.ThesisItem Open Access Bioherbicidal potential of antagonistic rhizosphere bacteria in management of wild oat (Avena fatua L.) and their inoculation effect on growth of wheat (Triticum aestivum L.)(CCSHAU, 2018) Dahiya, Anupma; Sindhu, S.S.In the present study, seventy five rhizobacterial isolates were obtained on the basis of morphological characteristics from wheat rhizosphere soil. These rhizobacterial isolates were screened for antagonistic interaction along with 13 reference strains against Rhizoctonia solani and Neovossia indica under in vitro conditions. Twenty three isolates showed antagonistic activity against R. solani and 33 isolates showed antagonistic activity against N. indica. Isolates BWA36, RWA42, RWA48, RWA53, HCA3, HCA61 and RCA3 showed significant growth inhibition against R. solani, whereas isolates BWA6, BWA19, BWA23, RWA48, RWA53, HCA61 and RCA3 showed the inhibition against N. indica. Thirty four selected antagonistic bacteria were studied for their effect on root and shoot growth of Avena fatua seedlings on 0.8% water agar plates. Eight rhizobacterial isolates BWA6, BWA19, BWA29, RWA48, RWA55, RWA63, RWA71 and HCA61 showed root growth inhibition at both 5th and 10th day of seed germination. Bacterial isolates i.e. BWA6, BWA19, BWA29, BWA38, RWA48, HCA61 and JMM24 caused shoot growth inhibition at both 5th and 10th day of seed germination of A.fatua. Culture filtrate of selected cultures were sprayed on 1-week and 2-weeks old Avena fatua and wheat seedlings. Isolates BWA25, BWA29, RCA3 and SYB101 caused yellowing of leaves, whereas two rhizobacterial isolates BWA18 and RWA52 caused appearance of disease spot on weed. IAA production in selected rhizobacterial isolates and it varied from 3.49 to 53.80 μg/ml. Significant ALA production (> 7 μg/ml) was observed in bacterial isolates BWA25, HCA61, RCA3, HCA3 and SYB101. ACC utilization was observed in 85.29% bacterial isolates and five isolates i.e., BWA20, BWA23, BWA29, BWA38 and RCA3 showed significant growth on ACC supplemented plates. Four bacterial isolates BWA25, BWA52, RWA53 and RCA3 showed significant HCN production. Ten bacterial isolates i.e., BWA2, BWA18, BWA25, BWA29, RWA48, RWA52, RWA69, SYB101, HCA61 and RCA3 were selected on the basis of different beneficial properties to study their inoculation effect on growth of wheat and weed under pot house conditions. Rhizobacterial isolates BWA18, RWA52, RWA69 and SYB101 stimulated growth of wheat and rhizobacterial isolates i.e., BWA18, BWA29 and RWA52 inhibited the growth of A. fatua in comparison to RDF amended uninoculated soil treatment. At 25 days of observation, bacterial isolate BWA18, RWA69, SYB101 showed significant increase in root dry weight (RDW) and shoot dry weight (SDW) of wheat, whereas its inoculation decreased RDW and SDW of A. fatua. At 50 days of observation, inoculation of bacterial isolates BWA18 and RWA48 increased RDW and SDW of wheat and weed both. Isolates RWA69 and SYB101 showed significant increase in RDW and SDW of wheat, whereas its inoculation decreased RDW and SDW of A. fatua. At 75 days of observation, inoculation with bacterial isolates RWA52, RWA69 and SYB101 caused significant increase in RDW and SDW of wheat, whereas its inoculation decreased RDW and SDW of A. fatua as compared to RDF amended uninoculated soil. Rhizobacterial isolates BWA18, RWA52 and RWA69 were identified as Acinetobacter variabilis, Bacillus siamensis and Bacillus endophyticus, respectively by the 16S rRNA sequence analysis. These bacterial isolates could be further exploited as bioherbicide for wild oat and growth improvement of wheat under field conditions.ThesisItem Open Access Biological control of subterranean termites (Isoptera: Termitidae) with soil bacteria(CCSHAU, 2014) Dua, Seema; Sindhu, S.S.Selected ten bacterial strains having termite killing ability along with two control strains was studied for antagonistic interactions with local soil termites. Live bacterial cells, heat killed cells and culture supernatants were tested for termiticidal activity. Different bacterial strains showed more than 80% killing of termites at 5 days of incubation. Four bacterial strains KBM79, KPM35, PPM147 and PBM195 caused 100% killing at 10 days of observation. The cell free culture filtrate studies of these antagonistic cultures showed that antagonistic substance is extracellular. Termite mortality percentage increased when culture supernatants were used as compared to whole cell broth of bacterial strains. The killing frequency of the culture supernatants obtained from different bacterial strains varied from 78.0 to 96.0% at 2nd day of observation. Termicidal activity was lost on treatment of whole cell broth and culture supernatant of bacterial strains by incubation at high temperature (1000C for 10 minutes). Bacterial strains KBM79 and KPM35 possessed proteolytic, lipolytic and chitinolytic enzyme activities and caused 100% killing of termites at 10 days. Partial purified proteins caused 100% killing of termites within half an hour. The SDS–PAGE analysis of selected bacterial strains showed four common protein/polypeptides having molecular weight 129 KDa, 37.1 KDa, 28.7 KDa and 26.3 KDa in all the ten bacterial strains. These proteins/polypeptides were missing in control bacterial strains WPS73 and KPM31. Plasmid-cured strain KPM35 lost the protein band having molecular weight 129 KDa. Termiticidal activity was also decreased in plasmid-cured strain KPM35 indicating that some of the termiticidal genes were located on the plasmid. Moreover, crude preparation and partial purified protein of bacterial strains KBM79 and KPM35 when treated with proteinase K lost the antagonistic activity indicating that extracellular protein could be involved in termite mortality. Combination of different chemical compounds with efficient bacterial strains showed high termite mortality than individual insecticides. Application of bacterial strain KPM35 and dexamethasone (50 ug/ml) caused 100% mortality even at 12 hours of incubation. Dexamethasone alone was found more lethal than boric acid and sodium citrate, and caused 90% mortality at 48 hours of incubation. Bacterial strains KBM79 and KPM35 were identified as Bacillus subtilis and Pseudomonas synxantha. These bacterial strains could be further tested for termiticidal activity under field conditions.ThesisItem Open Access Characterization of antagonistic rhizobacteria having plant growth promoting effects on mustard (Brassica juncea L.)(CCSHAU, 2015) Sharma, Ruchi; Sindhu, S.S.Indian mustard (Brassica juncea L.) is an important oilseed crop grown across the Northern Indian plains. The major constraints in growing mustard are diseases, aphid pests, weeds and abiotic stresses which are responsible for reduction in the growth, yield and oil production. Alternaria blight disease caused by Alternaria brassicae is the important disease of Indian mustard, which causes 10-70% yield losses. Fungicides mancozeb and iprodione are applied to control this disease. However, increasing awareness about environment pollution has emphasized the need for use of biocontrol agents for substituting pesticides. In the present study, seventy-eight bacterial isolates were obtained from the mustard rhizosphere at 50 and 75 days of plant growth and 305 isolates were procured from the Department of Microbiology, CCS H.A.U., Hisar. Out of 383 isolates tested, 20.88% isolates inhibited the growth of fungi Alternaria brassicae. Highest antagonistic activity (halo zone to growth ratio) was shown by isolate HMR25 (5.03) followed by HMR33 (4.68) and HMR70 (4.12). On the basis of variation in antagonistic activity, 46 isolates were further tested for production of indole-3-acetic acid (IAA), δ- aminolevulinic acid (ALA) and potassium solubilization activity. Twelve isolates showed 5-15 μg/ml IAA production and maximum IAA production was observed in isolate HMR57 (19.95 μg/ml) followed by HMR39 (17.99 μg/ml) and HMR2 (16.45 μg/ml). Maximum ALA production was observed in isolate WSA56 (15.62 μg/ml) followed by HMM21 (14.74 μg/ml) and HMM49 (14.45 μg/ml). Highest potassium solubilizing index (KSI) was observed in isolate HMR56 (3.50) followed by HMR73 (3.46) and HMR29 (3.25). Five antagonistic isolates HMR70, JMM16, HMR48, WHA64 and HMR25 were further selected for growth promotion and disease control on mustard crop under pot house conditions. At 25 days after sowing (DAS), isolate HMR70 showed 77.5% increase in shoot dry weight followed by JMM16 (47.5%) whereas 100% disease control was observed with HMR25, HMR48 and HMR70. At 50 DAS, JMM16 showed 161.1% increase in shoot dry weight followed by HMR70 (136.1%) whereas 100, 80.0 and 80.0% disease control was observed with isolates HMR25, HMR48 and WHA64, respectively. At 75 DAS, isolate HMR70 showed 114.6% increase in shoot dry weight followed by HMR48 (77.5%) and JMM16 (55.0%) whereas 100, 83.33 and 83.33% disease control was observed in HMR25, HMR48 and HMR70. Single inoculation of isolates HMR70 and JMM16 showed significant increase in shoot dry weight as compared to control. Coinoculation of rhizobacterial isolates HMR70, HMR48 and HMR25 with A. brassicae caused 83.33, 83.33 and 100.0% disease control, respectively at 75 days of plant growth. Rhizobacterial isolates HMR70, JMM16 and HMR25 showed more stimulation of mustard growth and also suppressed the disease. These three rhizobacterial isolates could further be assessed for their plant growth promotion and disease control potential under field conditions for their subsequent use as efficient biofertilizer and biocontrol agents.ThesisItem Open Access Evaluation of herbicidal potential of rhizosphere bacteria against bathu (Chenopodium album) and piazi (Asphodelus tenuifolius) weeds(CCSHAU, 2015) khandelwal, Aakanksha; Sindhu, S.S.Wheat (Triticum aestivum L.) is the most important cereal crop for the majority of world’s populations. Weeds cause significant losses each year in the agriculture crops. Instead of using herbicide, biological control is a promising approach for the effective weed control. Two hundred fifty rhizosphere bacteria were isolated from the rhizosphere of wheat and mustard. Among 250 isolates tested, ninety six rhizobacterial isolates showed significant stimulation or retardation effect on seed germination of weed Chenopodium album and Asphodelus tenuifolius on 0.8% water agar plates. Forty five isolates showed root growth inhibition on 5th day of seed germination in C. album. Nine rhizobacterial isolates caused shoot growth inhibition on 5th day and seven bacterial isolates caused shoot growth inhibition at 10th day of seed germination of C. album. In Asphodelus tenuifolius, thirty four isolates showed root growth inhibition on 5th days and twenty seven rhizobacterial isolates showed root growth inhibition at 10th day of seed germination. Eight rhizobacterial isolates caused shoot growth inhibition on 5th day and twenty four rhizobacterial isolates caused shoot growth retardetion at 10th day of seed germination. Screening of Ninety six rhizobacterial isolates for production of indole acetic acid showed that ninety six per scent isolates produced IAA ranging from 0.21 to 29.91μg/ml. Rhizobacterial isolates WSA38, MSA57, WSA68, WSA56, MSA42, MSA39, WHA98 and MSA11 showed >11.0 μg/ml production of δ-aminolevulinic acid and eighty per cent isolates produced ALA. Sixty three per cent of rhizobacterial isolates showed growth on ACC supplemented plates. The ability to produce siderophore was found only in 30% of the isolstes and twenty nine bacterial isolates showed HCN production. Eight bacterial isolates i.e., MSA39, MSA56, MHA75, MHA93, WSA56, WHA82, WHA87 and WHA100 were tested for their effect on growth of wheat and weed under pot house conditions. Inoculation of bacterial isolate WHA87 showed 94-182% increase in root dry weight (RDW) and 30-340% increase in shoot dry weight (SDW) of wheat, whereas its inoculation caused 21-81% decrease in RDW and 33- 43% decrease in SDW of Chenopodium album at 30, 60 and 90 days of plant growth. Similarly, inoculation with bacterial isolate MSA39 caused 20.5-66% increase in RDW and 12-23.5% increase in SDW of wheat growth and its inoculation caused 14-89% decrease in RDW and 6-16% decrease in SDW of C. album. In case of Asphodelus tenuifolius, inoculation of bacterial isolate MSA56 showed 94-368% increase in RDW and 38-412% increase in SDW of wheat, whereas its inoculation caused 40- 85.7% decrease in RDW and 53-54.3% decrease in SDW of A. tenuifolius. Similarly, inoculation with bacterial isolate MHA75 caused 77-132% increase in RDW and 21-231% increase in SDW of wheat growth and its inoculation caused 25-75% decrease in RDW and 33-44% decrease in SDW of A. tenuifolius at 30, 60 and 90 days of plant growth. Thus, rhizobacterial isolates i.e., WHA87, MSA39, MHA75 and MSA56 were found to stimulate growth of wheat, whereas isolates i.e., MSA39 and WHA87 inhibited the growth of Chenopodium album and isolates MHA75, MHA93 and MSA56 inhibited the growth of Asphodelus tenuifolius.ThesisItem Open Access Fusarium wilt management and growth stimulation of chickpea (Cicer arietinum L.) by inoculation with antagonistic Bacillus species(CCSHAU, 2017) Sehrawat, Anju; Sindhu, S.S.In the present study, one hundred ninety three Bacillus isolates were obtained from chickpea rhizosphere soil and screened for their antagonistic interactions against Fusarium oxysporum under in vitro conditions. Eighty eight isolates showed antagonistic activity against F. oxysporum and 24 isolates were selected. Bacillus isolates HCA3, HCA26, HCA53, HCA61, RCA3 and RCA9 showed maximum inhibition of F. oxysporum growth. Nine Bacillus isolates also inhibited the growth of Aspergillus niger, Neovossia indica and Rhizoctonia solani. Siderophore production was observed in four isolates HCA1, HCA3, HCA53 and RCA11. ACC utilization was observed in all selected isolates except RCA9. Three Bacillus isolates RCA3, RCA7 and RCA11 showed significant HCN production. Significant ALA production (> 12 μg/ml) was observed in seven Bacillus isolates. The IAA production varied from 0.13 to 29.60 μg/ml in different isolates. Thirteen Bacillus isolates showed phosphorus solubilizing activity and nine isolates caused potassium solubilization. Three Bacillus isolates i.e. RCA4, RCA9 and RCA19 retarded the growth of chickpea seedlings on 0.8% soil extract agar plates, whereas other isolates showed growth stimulation effect on seedling growth in comparison to uninoculated control at both 5th and 10th day of growth. Out of 24 Mesorhizobium isolates obtained from chickpea nodules, eight isolates showed antagonistic effect against F. oxysporum. Bacillus and Mesorhizobium isolates were tested for their salt tolerance capacity at 1, 2, 4, 6 and 8% NaCl concentrations. Six isolates showed growth of 10.1 mm to 15.0 mm colony size at 6% NaCl concentration and five Bacillus isolates i.e. HCA11, HCA81, HCA53, RCA11, RCA31 showed larger colony size ranging from 5.1-10.0 mm even at 8% NaCl concentration. Only two Mesorhizobium isolates MCA5 and MCA22 were found salt-tolerant upto 8% of salt concentration. Maximum increase in SDW (45.5%) was observed by inoculation of isolate MCA20 followed by 44.3% increase in SDW by inoculation of isolate MCA23 at 40 days of chickpea growth under chillum jar conditions, whereas isolates MCA23 and MCA4 resulted in 166.2 and 161% increase in RDW, respectively. Likewise, 112.6% increase in SDW was observed on inoculation of MCA14 isolate followed by 118.3 and 117.2% increase in SDW on inoculation of MCA4 and MCA10 isolates, respectively at 80th day of observation. The inoculation effect of three antagonistic i.e., HCA61, RCA3 and RCA7, and two non-antagonistic Bacillus isolates HCA76 and SYB101 was studied with Mesorhizobium strains MBD26 and KR48 on chickpea growth. At 90 days after showing (DAS), coinoculation of MBD26+RCA3 and KR48+RCA7 showed significant increase in root and shoot dry weight of chickpea under chillum jar conditions. Bacillus isolates i.e. HCA61, RCA3 and RCA7 coinoculated with Mesorhizobium strains showed 100% disease control of F. oxysporum at different stages of chickpea growth under pot house conditions. At 60 DAS, coinoculation of F. oxysporum+KR48+SYB101 showed significant increase (155.4%) in SDW followed by coinoculation of F. oxysporum+KR48+RCA7 (124%) and F. oxysporum+MBD26+RCA7 (121.1%), whereas at 75 DAS, 173.3% increase in SDW followed by 130.9 and 128.3% increase was observed in these treatments under pot house conditions. At 90 DAS, coinoculation of F. oxysporum+KR48+SYB101 caused 149 and 111% increase in SDW and RDW of chickpea followed by F. oxysporum+KR48+RCA7 (129.5, 96.9%) and F. oxysporum+KR48+RCA3 (107.7, 77.2%), respectively. Bacillus isolates RCA3 and RCA7 could further be assessed for their plant growth promoting and disease control potential under field conditions for their subsequent application as biofertilizer and biocontrol agents.ThesisItem Open Access Germination and transplanting studies in aonla (Emblica officinalis gaertn.)(CCSHAU, 2006) Rashmi Kumari; Sindhu, S.S.The present investigation was carried out at the experimental orchard of Department of Horticulture CCS Haryana Agricultural University, Hisar. In the experiment, the effect of sowing time (3rd week of March and 3rd week of April) and chemicals (GA3 – 250, 500, 750 ppm; Thiourea 0.5, 1.0, 1.5% and KNO3 – 0.5, 1.0, 1.5%) on seed germination in aonla and the transplanting of budded aonla plants with earth ball and without ball during 1st week of July, August, September, January, February and March were studied During the present investigation 500 ppm GA3 was found to be the most effective treatment resulting in significant increase in seed germination (75.50%), survival percentage (83.37%), seedling height (72.94 cm), seedling girth (0.63 cm), number of branches per plant (3.13 cm), percent buddable seedling (80.44%) and early germination (8.0 days) followed by 1.0% thiourea. The seeds soaked in water also gave better performance as compared to untreated seeds. The 3rd week of March was found to be the best time of sowing of seeds as compared to 3rd week of April sowing. The transplanting of budded aonla plants with earth ball was found to be the best treatment as compared to plants without earth ball in all the months. The maximum survival percentage was obtained in plants with earth ball transplanted during 1st week of February (80.00%) followed by 1st week of July (73.30%), whereas, plants without earth ball gave maximum survival percentage when transplanted was done during 1st week of February (73.30%).ThesisItem Open Access Growth stimulation of pigeon pea (Cajanus cajan L.) by using antagonistic rhizospheric bacteria(CCSHAU, 2019) Sharma, Ruchi; Sindhu, S.S.Pigeon pea (Cajanus cajan L.) is one of the important grain legume crop in the developing countries of the tropical and subtropical regions of the world. It is an excellent source of protein (20-22%), supplementing energy rich cereal diets in a mainly vegetarian population. One of the major factors adversely affecting pigeon pea productivity is poor nodulation in fields due to prevalence of poor nodulating native rhizobia. Moreover, the crop productivity of pigeon pea is also adversely affected by biotic stresses such as Fusarium wilt (FW), Alternaria blight and Rhizoctonia root rot disease. In the present study, 83 rhizobacterial isolates and 24 rhizobial isolates were isolated from different soil samples. Screening of the isolates for antagonistic activity against three Alternaria tenuissima, Fusarium udum and Rhizoctonia bataticola showed that only 21% of the rhizobacterial isolates inhibited the growth of all the three pathogenic fungi. Four isolates i.e., HPR6, HPR22, HPR71 and HPR78 showed more than 4.0 halo zone to growth ratio against Alternaria tenuissima wheareas three isolates i.e., HPR37, HPR41 and HPR42 showed more than 4.0 halo zone to growth ratio against Fusarium udum and 3 isolates i.e., HPR5, HPR71 and HPR78 showed more than 4.0 halo zone to growth ratio against Rhizoctonia bataticola. Out of the 24 rhizobial isolates tested only 2 isolates i.e., RPR17 and RPR24 showed inhibition activity against all the three pathogenic fungi. Bradyrhizobial isolate RPR16 showed maximum halo-zone to growth ratio i.e., 4.78 against Fusarium udum. Five rizobacterial isolates i.e., HPR49, HPR77, HPR80, HPR83 and HPR17 showed significant ACC utilization activity. Eight isolates i.e. HPR54, HPR58, HPR59, HPR65, HPR67, HPR71 and HPR83 produced IAA ranging from 12.3-26.31 μg/ml. Rhizobacterial isolates showed ALA production in the range of 0.1-23.45 μg/ml. Potassium solubilization studies showed that 28 isolates showed significant activity in the range of 4.1-6.0 and 42 rhizobacterial isolates showed KSI in the range of 2.1-4.0. The ability of siderophore production was found in only 41% of the rhizobacterial isolates. Significant cyanide production was observed in isolate HPR40 i.e., 76.54 μl/ml. Among the 83 rhizobaterial isolates, only six isolates i.e., HPR1, HPR36, HPR39, HPR15, HPR48 and HPR69 showed growth up to 8% NaCl salt concentration, whereas 26 isolates showed growth up to 4% salt with different colony size. Maximum increase in SDW (101.85%) was observed by co-inoculation of isolates RPR17+HPR78 followed by 97.68% increase in SDW by inoculation of isolate RPR17+HPR78 at 45 days of pigeon pea growth under chillum jar conditions. Coinoculation of isolates RPR17 + HPR78 and RPR17 + HPR17 resulted in 86.95 and 83.18% increase in RDW, respectively. Likewise, 108.59% increase in SDW was observed on inoculation of RPR17 + HPR78 isolates followed by 118.3 and 94.05% increase in SDW on inoculation of RPR17+HPR17 isolates, respectively at 60th day of observation. The inoculation effect of four antagonistic i.e., HPR17, HPR40, HPR49 and HPR78, and two bradyrhizobial isolates i.e., RPR19 and MRH46 was studied under pot house conditions on pigeon pea growth. At 45 DAS, the coinoculation treatment of RPR19 + HPR40 alongwith F. udum showed maximum nodulation i.e., 26 nodules/plant and 79.59 and 98% increase in SDW and RDW, respectively. At 60 DAS, the coinoculation treatment of RPR19 + HPR17 showed maximum nodulation i.e., 31 nodules/plant and 37.10 and 21.70% increase in SDW and RDW, respectively. At 75 DAS, coinoculation of RPR19 + HPR40 showed maximum nodulation i.e., 52 nodules /plant and showed significant increase in root and shoot dry weight of pigeon pea under pot house conditions. Rhizobacterial isolates HPR78 showed 100% disease control of F. udum at different stages of pigeon pea growth under pot house conditions. Coinoculation treatments i.e., RPR19 + HPR49 showed 100% disease control of Fusarium wilt at different stages of pigeon pea growth under pot house conditions. Rhizobacterial isolates HPR40 and HPR78 were identified as Pseudomonas aeruginosa strain SSR207 and Paenibacillus polymyxa strain SSR2_207. These plant growth promoting rhizobacterial isolates can be used for the enhancement of crop productivity under salinity stress in field conditions.ThesisItem Open Access Indole acetic acid production by Pseudomonas species and its effect on plant growth of green gram and black gram(CCSHAU, 2004) Om Prakash; Sindhu, S.S.In the present studies, IAA producing Pseudomonas strains CPS59 and MPS90 were mutagenized with transposon Tn5 using E. coli strain S17-1. The frequency of mutants with low amount of IAA production varied from 35.14 to 46.75% from Pseudomonas strains CPS59 and MPS90, respectively and only 3.43 to 3.75% mutants produced higher levels of IAA in comparison to respective parent strains in LB medium supplemented with tryptophan. Inoculation of mutants derived from Pseudomonas strain CPS59 showed stunting effect on root development of green gram seedlings at both 5 and 10 days of observation whereas mutants CPS59-138, CPS59-162 and CPS59-231 showed stimulatory effect on root growth at 5 days in black gram. Majority of the mutants caused stimulation of shoot growth at both 5 and 10 days of observation in comparison to uninoculated treatment. MPS90-derived mutants i.e. MPS90-39, MPS90-157, MPS90-102 and MPS90-106 showed slight stimulation of root growth whereas most of the mutants showed stunting effect on shoot growth at 10 days in black gram. Coinoculation studies of Pseudomonas mutants with Bradyrhizobium sp. strain S24 resulted in increased nodule number, nodule fresh weight and shoot dry weight in green gram and black gram under chillum jar conditions. Shoot dry weight gains in green gram after coinoculation with CPS59-derived mutants varied from 107-149 per cent and from 110-137 per cent with MPS90-derived mutants in comparison to Bradyrhizobium-inoculated plants at 60 days of growth. Similarly, shoot dry weight gains in black gram varied from 102-178 per cent and 105-198 after inoculation with mutants derived from CPS59 and MPS90, respectively. The stimulation effect on shoot dry weight in comparison to uninoculated control treatment varied from 280-390 per cent in green gram and 179-357 per cent in black gram. Four mutants CPS59- 138, CPS59-321, MPS90-133 and MPS90-51 showed more enhancements in nodule formation by Bradyrhizobium strain S24 in green gram whereas mutants CPS59-162, CPS59-64, MPS90- 102 and MPS90-280 caused more stimulation for nodule formation in black gram at 60 days of plant growth. Significant enhancements in shoot dry weights of green gram were observed by coinoculation with Pseudomonas mutants CPS59-138, CPS59-231, CPS59-321, MPS90-280, MPS90-14 and MPS90-145. Similarly, coinoculation of Bradyrhizobium strain with Pseudomonas mutants CPS59-162, CPS59-321, CPS59-214, MPS90-102, MPS90-280 and MPS90-51 showed significant gains in shoot biomass of black gram at 60 days of plant growth. The better performance of IAA over producing mutants in relation to nodulation and plant biomass indicated that IAA production by Pseudomonas strains is a beneficial trait for selection of rhizobacterial strains having plant growth promotion ability.ThesisItem Open Access Isolation and identification of bacteria having pathogenic interactions with termites (Isoptera)(CCSHAU, 2007) Yuvraj singh; Sindhu, S.S.In the present study, the population of bacteria in termite nest soil varied from 1.2 x 106 to 90.0 x 106 colony forming units/g soil on nutrient agar medium. On soil extract agar medium, the bacterial population varied from 28.3 x 106 to 103.2 x 106 cfu/g soil and it varied between 2.6 x 106 to 256.0 x 106 cfu/g soil on King’s B medium. Based on colony morphology, gum production, colony characteristics and pigment production, bacterial isolates NNY 1-NNY 84 were selected from nutrient agar medium, isolates NSY 1-NSY 40 from soil extract agar medium and isolates NKY 1-NKY 96 were selected from King’s B medium. From the gut of termites, bacterial isolates GNY 1-GNY 20 were selected from nutrient agar medium plates. Isolates GSY 1-GSY 10 were obtained from soil extract agar medium and isolates GKY 1-GKY 20 were obtained from King’s B medium. A total of 270 bacterial isolates, i.e. 220 isolates from termite nest soil and 50 isolates from termite gut, were screened for the potential to produce different enzymes, i.e. lipase, protease and chitinase on specific media. In the preliminary screening, only 83 isolates were found to express one or more of the enzyme activities. Seven bacterial isolates without any enzyme activity were also selected. For lipolytic activity, the clearance zone to colony growth ratio varied from 1.24 to 4.20 in different bacterial isolates. Bacterial isolates NNY 73 and NSY 19 showed very little lipase activity and eight isolates did not show any lipase activity. Three isolates NNY 19, NNY 39 and NSY 20 showed 3.50, 4.20 and 4.00 enzyme production efficiency, respectively. Proteolytic enzyme production in different cultures varied from 1.24 to 2.29. The bacterial isolates NNY 24, NNY 50 and NNY 62 showed highest ratio of clearance zone to colony size i.e. 2.17, 2.14 and 2.29, respectively. Fourteen bacterial isolates did not show any proteolytic activity. Chitinolytic activity varied from 1.24 to 2.67 in different bacterial isolates. Isolates NNY 18, NNY 52, NNY 73 and GKY 10 showed very little chitinolytic activity. Four isolates, NNY 23, NNY 43, NNY 34 and NKY 91 showed 2.67, 2.50, 2.40 and 2.50 enzyme production efficiency, respectively. The bacterial isolates NNY 58, NKY 48 and NKY 69 expressed only chitinolytic activity. Isolate NKY 66 expressed only proteolytic activity whereas isolates NKY 17, NKY 62 and NKY 79 expressed only lipolytic activity. Twelve bacterial isolates were found to express all three enzyme activities. During studies of pathogenic interactions, only 47 bacterial isolates were found to kill the termites at 5 day of observation. The killing frequency of different bacterial isolates varied from 5.7 to 100 per cent at 5 day. Bacterial isolates NNY 23, NSY 19 and NKY 83 caused 100 per cent killing of the termites whereas 14 other isolates caused more than 82 per cent killing at 5 day. At 7 days of observation, 100 per cent killing of the termites was observed with four more bacterial isolates NNY 19, NNY 43, NSY 3 and NKY 91. Bacterial isolate NNY 23 possessed all the three enzyme activities and caused 100 per cent killing of the termites at 5 day of observation. Another bacterial isolate NKY 64 which did not show any of the enzyme activity but it caused 57.8 per cent killing of termites at 5 day. These results indicated that besides the production of three enzymes, some other metabolites (toxin or siderophore) could also be contributing to the killing of termites. Reisolation of bacteria from dead termites showed that seven bacterial isolates viz. NNY 52, NNY 62, NSY 3, NSY 10, NKY 25, NKY 27 and NKY 31 killed even the gut bacteria of the termites and only colonies of inoculated pathogenic bacteria appeared on nutrient agar, soil extract agar and KB medium plates. In the study of other termites inoculated with other 16 bacterial isolates, more than two types of colonies were observed on medium plates indicating that gut bacteria were not killed after ingestion/infection of antagonistic bacteria. Twenty-one bacterial isolates having more than 80 per cent killing of termites at 7 day of observation were identified by standard procedures up to genus level as described in Bergey’s Manual of Determinative Bacteriology. Gram-positive staining was observed only in bacterial isolates NNY 10, NNY 34, NNY 50, NNY 52, NSY 2, NSY 3, NSY 19, NKY 83, NKY 91 and GNY 17. Based on the comparative analysis of various morphological (colony morphology, pigment production and sporulation) and biochemical characteristics (such as indole production, methyl red test, Voges-Proskauer reaction, citrate utilization, oxidase, catalase and hydrolysis of starch and cellulose), the antagonistic bacteria were found to belong to the genera of Arthrobacter, Bacillus, Chromobacterium, Enterobacter, Micrococcus, Neisseria, Pseudomonas and Serratia.ThesisItem Open Access Isolation of potassium solubilizing bacteria and their inoculation effect on growth of wheat (Triticum aestivum L. em. thell.)(CCSHAU, 2011) Parmar, Priyanka; Sindhu, S.S.In the present study, seventy bacterial isolates obtained from wheat rhizosphere and 67 reference strains from the department were tested for potassium solubilization ability on Aleksandrov medium supplemented with mica as potassium source. Twenty isolates/strains solubilized potassium from mica and the amount of K released by the isolates/strains ranged from 15 mg/L to 48 mg/L. Maximum K solubilization occurred with glucose as carbon source, at 25°C temperature of incubation, 7.0 pH of the medium and with KCl as potassium source followed by K2SO4. Isolate HWP47 caused solubilization of potassium in mica only by acid production, isolates HWP28 and HWP69 by production of CPS and EPS, HWP38 by production of acid and CPS whereas isolates HWP15, HWP17, HWP53, HWP 57, HWP61 and HWP63 caused solubilization by production of acid, CPS and EPS. Inoculation of K solubilizing isolate HWP47 in wheat (Triticum aestivum L.) variety WH711 caused 51.46% increase in RDW in soil and 60.19% increase when rock material was added in pots at 60 days after sowing. Similarly, 44.28% increase in SDW was found in HWP47 inoculated plants. At 90 days after sowing, the inoculation of rhizobacterial isolate HWP15 caused 15.29% and 27.19% increase in RDW and SDW, respectively. The plant dry weight gain was further enhanced to 20.59% and 71.92% in RDW and SDW, respectively with amendment of rock material. Inoculation with isolate HWP47 showed 22.35% increase in RDW and 73.68% increase in SDW on addition of rock material. Isolates HWP15 and HWP47 also caused significant K uptake in the shoot tissues. Thus, potassium solubilizing bacteria HWP15 and HWP47 could be further exploited for plant growth improvement under field conditions.ThesisItem Open Access Mulching and weed control studies in guava (Psidium guajava Linn.)(CCSHAU, 2014) Deepak, Singh; Sindhu, S.S.The experiment entitled “Mulching and Weed Control Studies in Guava” was conducted at Research Farm of the Department of Horticulture, CCS Haryana Agricultural University, Hisar during 2011-13 in rainy and winter season to find out the suitable mulching and herbicides on moisture status, weed control, nutrient content in soil and plants as well as improvement in growth, yield and quality of guava. The cultivar used for the investigation was Hisar Safeda. The treatments comprising six levels of mulches and nine levels of herbicides were laid out in a randomized block design with four replications keeping a net plot size of 6.0 x 6.0 m separately. The data recorded on various parameters, which were influenced by different levels of mulching and herbicides. In mulching experiment, the minimum value for soil pH and ECe, while, maximum values for soil moisture per cent, NPK and growth parameters viz. plant girth, spread, height, shoot length, and leaf water potential were recorded with black polythene of 100μ mulch which was at par with paddy straw mulch. However, maximum leaf NPK, TSS, ascorbic acid and minimum acidity were recorded in paddy straw mulch, while, maximum fruit weight, length, breadth and yield were recorded under black polythene of 100μ mulch which was statistically similar with paddy straw mulch. Minimum weed counts, fresh and dry weight of weeds at all monthly intervals were observed with black polythene of 100μ mulch during both the years in rainy and winter season. The two year results suggest that the paddy straw mulch has shown the best economical mulch for higher yield with quality fruit production at commercial scale. In weed control experiment, soil pH, ECe, leaf NPK, water potential and fruit acidity were remain unchanged during the time frame. However, manual weeding at monthly interval treatment found best for all other parameters viz., soil NPK, moisture, growth, yield, quality and weed parameters while, in herbicidal treatments, atrazine 2 kg + glyphosate 1% recorded best for the same. Manual weeding at monthly interval treatments gave the best results but it was very costly and herbicidal combination of atrazine 2 kg + glyphosate 1% resulted almost similar to manual weeding at monthly interval treatment. Atrazine 2 kg + glyphosate 1% found best economical combination for commercial production of guava with quality fruits. Paddy straw mulch and in herbicidial treatments, atrazine 2 kg + glyhosate 1% found best treatments for guava orchard in respect to all above characters under semi arid condition of Hisar (Haryana).ThesisItem Open Access Mulching and weed control studies in Kinnow(CCSHAU, 2014) Bhanukar, Manoj; Sindhu, S.S.The experiment entitled “Mulching and Weed Control Studies in Kinnow” was conducted at Research Farm of the Department of Horticulture, CCS Haryana Agricultural University, Hisar during the year 2013 to find out the suitable mulching and herbicides on moisture status, weed control and nutrient content in plants as well as improvement in growth, yield and quality of kinnow. The treatments comprising six levels of mulches and nine levels of herbicides were laid out in a randomized block design with three replications keeping a net plot size of 6.0 x 6.0 m separately. The data were recorded on various parameters, which were influenced significantly by different levels of mulching and herbicides. In mulching experiment, the minimum value for soil pH and ECe, while, maximum values for soil moisture per cent, NPK and growth parameters viz. plant girth, spread, height and leaf water potential were recorded with black polythene of 200μ mulch. However, maximum TSS, ascorbic acid and minimum acidity were recorded in paddy straw mulch and maximum juice content was recorded with black polythene 200μ mulch, while maximum fruit weight, length, breadth, number of fruits and yield were recorded in black polythene 200μ mulch. Similarly, lowest number, fresh and dry weight of weeds at all monthly interval were observed with Black polythene 200μ and 100μ mulch followed by paddy straw mulch. The results suggest that the Black polythene 200μ mulch has shown the best economical mulch for higher yield with quality fruit production at commercial scale. In weed control experiment, soil pH, ECe, leaf water potential and fruit acidity were remaining unchanged during the time frame. However, manual weeding at monthly interval treatment found best for all other parameters viz. soil NPK, moisture, growth, yield, quality and weed parameters while, in herbicidal treatments, atrazine 2 kg + glyphosate 1% recorded best for the same. Manual weeding at monthly interval treatments gave the best results but it was very costly and herbicidal combination of atrazine 2 kg + glyphosate 1% resulted almost similar to manual weeding at monthly interval treatment. Atrazine 2 kg + Glyphosate 1% found best economical combination for commercial production of kinnow with quality fruits under semi arid condition of Hisar (Haryana).ThesisItem Open Access Siderophore production by pseudomonas species and its role in disease control of green gram (Vigna radiata) caused by rhizoctonia solani(CCSHAU, 2006) Sahu, Govind Kumar; Sindhu, S.S.In the present study, sixty-five bacterial isolates were obtained from the rhizosphere soil of chickpea and green gram plants by plating serial dilutions on King’s B media. Fifty-eight isolates were identified as Pseudomonas based on colony morphology, pigment characteristics and oxidase test. These fifty-eight Pseudomonas isolates alongwith 22 reference strains were tested for siderophore production using universal chromo-azurol S assay method on MM9 medium and iron-deficient succinate medium. Thirty-seven Pseudomonas strains/isolates were found to produce siderophore on MM9 medium and the diameter of halo zone varied with different Pseudomonas isolates. Production of siderophores was found more on MM9 medium as compared to succinate medium. Bacterial colony growth (G) and halo zone (H) size were measured on MM9 medium and H/G ratios varied from 1.44 to 10.24 in different Pseudomonas isolates. Seventeen Pseudomonas isolates were used for optimization of cultural conditions for enhanced production of siderophores. Replacement of glucose with sucrose, sodium gluconate or sodium malate supported siderophore production whereas arabinose addition repressed siderophore production in most of the Pseudomonas isolates. Increased concentration of ammonium chloride (0.2%) also decreased siderophore production. Addition of Fe-EDTA salt at low 5 M concentration showed less suppressive effect whereas at 10 M concentration more inhibitory effect on siderophore production was observed. Production of siderophore was found more at pH 7.0 in comparison to cultures grown in media with pH 8.0 and pH 6.0. Siderophore producing Pseudomonas isolates inhibited the growth of phytopathogenic fungi Fusarium oxysporum, Rhizoctonia solani and Pythium aphanidermatum. Large inhibition zones of fungal growth were observed on PDA medium in comparison to NA medium plates. Coinoculation of Pseudomonas cultures caused reduction in root rot disease symptoms under pot house conditions and disease control varied from 33.4 to 100 per cent with different Pseudomonas cultures. Coinoculation of Pseudomonas isolates MP20, MPS54, CP56 and CPS67 with Bradyrhizobium strain SMR15 also enhanced the nodule number, nodule fresh weight and plant dry weight as compared to Bradyrhizobium-inoculated or uninoculated control plants at 30, 45 and 60 days of plant growth. Coinoculation of Pseudomonas strain CP56 with Bradyrhizobium strain and R. solani showed maximum increase (275.8%) in plant dry weight at 60 days in comparison to control plants and completely suppressed the root rot disease under pot house conditions. The better performance of siderophore-producing Pseudomonas strains in relation to symbiotic performance and disease control indicated that Pseudomonas strains could be exploited for disease suppression and plant growth promotion under field conditions.ThesisItem Open Access Studies On Processing Of Aonla Beverages And Its Keeping Quality(Chaudhary Charan Singh Haryana Agricultural University; Hisar, 2010) Lather, Rajesh; Sindhu, S.S.