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Dr. Y. S. Parmar University of Horticulture & Forestry, Solan
Dr. Yashwant Singh Parmar University of Horticulture and Forestry, Solan, was established on 1st December, 1985 with the objective to promote education, research and extension education in the fields of Horticulture, Forestry and allied disciplines. Late Dr. Yashwant Singh Parmar, the first Chief Minister and the architect of Himachal Pradesh perceived the importance of Horticulture and Forestry to develop and improve the State economy which led to the establishment of this University. Its history lies in erstwhile Himachal Agricultural College, Solan, established in 1962 and affiliated to the Panjab University. It became one of the campuses of Agriculture Complex of Himachal Pradesh University on its formation in 1970. Consequent upon the establishment of Himachal Pradesh Krishi Vishvavidyalaya in 1978, this campus became its Horticulture Complex and finally in 1985, assumed the status of a State University, being the only University in the country engaged exclusively in teaching, research and extension in Horticulture and Forestry.
The University is located at Nauni in Solan District of Himachal Pradesh, 13 km from Solan on Solan-Rajgarh Road, at an elevation of 1300 metres above mean sea level. Solan town is situated on national highway (NH-22) and is well connected by train and bus services.
The University has four constituent colleges, out of which, two are located at the main campus Nauni, one for horticulture and the other for forestry, having 9 and 7 departments, respectively. The third College i.e., College of Horticulture & Forestry is located at Neri in Hamirpur District on Nadaun-Hamirpur state highway, about 6 Km from Hamirpur town and is well connected with bus service. The college offers three Undergraduate Degree Programmes i.e. BSc (Hons.) Horticulture, BSc (Hons.) Forestry and B. Tech. Biotechnology and MSc degree programme in a few subjects. The fourth college i.e. College of Horticulture and Forestry, Thunag (Mandi) is located at Thunag District Mandi. This college offer BSc (Hons.) Horticulture and BSc (Hons.) Forestry degree programme. In addition, there are five Regional Research Stations, 12 Satellite Stations and five Krishi Vigyan Kendras (KVKs) situated in different zones of the State.
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ThesisItem Open Access STUDIES ON ANNEXIN GENE FAMILY FROM SOLANACEOUS VEGETABLES(UHF,NAUNI, 2019-12) SHARMA, ANKITA; NATH, AMARJIT K.ABSTRACT Heat stress leads to adverse effect on Potato cultivation affecting the tuberization and the bulking process. Being a signaling molecule, it is hypothesized that annexins might play important role in tuberization under heat stress condition. Ten annexins from potato and 13 annexins from tomato were identified using bioinformatics tool. Annexin gene proteins from potato and tomato were found to have an average molecular weight of 36kDa. Both potato and tomato annexins showed differential location of the annexin genes on 12 chromosomes. Expression profiling of Potato annexin genes under heat stress conditions revealed strong up-regulation of annexins in heat tolerant potato cultivar Kufri Surya as compared to susceptible one i.e Kufri Chandramukhi indicating their role as key signaling molecule under heat stress conditions. The annexin gene StAnn1 showed a uniform pattern of elevated gene expression in both leaves and stolon under heat stress throughout the developmental stage of potato cultivar KS as compared to potato cultivar KCM. Significant increase in peroxidase, anthocyanin and proline content was observed in heat tolerant cultivar Kufri Surya on different intervals of exposure to heat stress. The content of MDA was significantly less in Kufri Surya as compared to Kufri Chandramukhi indicating much less membrane damage in the tolerant potato cultivar KS. The annexin gene StAnn1 was isolated and cloned from Kufri Surya and over expressed in heat susceptible cultivar Kufri Chandramukhi for exploiting its potential for providing heat stress tolerance in other susceptible potato cultivars. Agrobacterium mediated genetic transformation protocol was standardized for potato cultivar Kufri Chandramukhi where selective MS medium with composition containing IAA 0.042mg/L, GA33.0 mg/L, Zeatin 3.0 mg/L, NAA 0.008 mg/L, Kan 50 mg/L, Carb 250 mg/L, Cefo 100 mg/L showed maximum percentage of callus formation i.e 76 % and shoot regeneration. The best rooting was observed on selective root regeneration MS medium containing 50mg/L Kan and 0.3mg/L IAA. On exposure to in vitro salt stress, transgenic KCM plants expressing StAnn1 annexin gene, phenotypically showed better tolerance as compared to the control plants depicting its potential in providing tolerance to other abiotic stress also. A decline in number of roots, nodes/plant and plant height was observed in control shoots as compared to putative transgenic shoots.ThesisItem Open Access IN VITRO MUTAGENESIS IN GINGER (Zingiber officinale Rosc.) FOR THE SELECTION OF PLANTS RESISTANT TO Fusarium oxysporum f.sp. zingiber(UHF,NAUNI, 2019-12) SHARMA, VISHAL; THAKUR, MANISHAABSTRACT In the present investigation, an attempt has been made to develop Fusarium yellows resistant plants of ginger (Zingiber officinale Rosc.) var. using in vitro mutagenesis and shoot selection technique. In vitro propagation was done using buds as explants. Maximum in vitro bud establishment was achieved in spring season (62.85%). 0.2% HgCl2 for 5 minutes was found to be the best concentration of surface sterilant with 42.59 per cent survival. 72.85% buds established on MS medium supplemented with 1 mg/l BA and 0.1 mg/l NAA. MS medium fortified with 0.5 mg/l BA and 0.1 mg/l NAA was found to be best multiplication medium resulting in 5.2 average number of shoots per explant with average shoot length of 4.5 cm. Rooting could be achieved on the same medium after second subculture. With the increase in number of subcultures, shoot multiplication rate, average shoot length, per cent rooting, thickness of roots and average number of roots per shoot showed an increase. In vitro cultures were subjected to gamma irradiation (10-100 Gy) for in vitro mutagenesis. LD50 was calculated after every four weeks and it was calculated to be 15 Gy after 16 weeks of irradiation. The non irradiated and irradiated plantlets were hardened with 100 per cent survival rate in potting mixture comprising of soil: cocopeat: FYM (2:1:1). On biochemical evaluation of rhizomes, it was observed that essential oil content was found to be same in all the samples (2%) whereas, highest oleoresin content was found in tissue culture propagated rhizomes (3.16%) and lowest crude fibre content (5.3%) was obtained in rhizomes collected from gamma irradiated plants. HPLC studies on gingerol content showed its highest value in rhizomes produced by gamma irradiated plants. No difference in chromosome number was observed in tissue culture propagated and gamma irradiated shoots. Gamma irradiated shoots were cultured on selective medium containing different concentrations (0-20%) of fungal culture filtrate (FCF) for in vitro selection. 17.5% FCF was found to be the highest concentration on which 5% shoots survived after 1st selection cycle which reduced to 1.15% after 3rd alternate, discontinuous selection cycle. Selected shoots from 15 and 17.5% FCF were multiplied on multiplication medium. Rooted plantlets were successfully hardened with 100% survival. 46.4 and 52% plants selected at 15 and 17.5% FCF were found to be highly resistant on in vivo evaluation. SCoT and SSR analysis distinguished between FCF selected, tissue culture propagated and gamma irradiated plants. In SSR analysis, two unique bands were obtained in plantlets selected on 17.5% FCF. On sequencing, the band obtained with GIN6 showed 98% homology with disease resistance protein-like gene CC-NBS-LRR from clone ZwP627 and band obtained with primer GIN9 showed 97% homology with same gene from clone ZoP620 of Z. officinale.After in vivo bioassay, SSR analysis of selected highly resistant plants again confirmed the unique bands with both the primers and the unique DNAsequences obtained have been submitted to NCBI under accession number MN497252 and MN497253 and will be published in GenBank after complete processing. Therefore, in the present study we have successfully developed Fusarium yellows resistant plants of ginger (Zingiber officinale Rosc.) var. Himgiri using in vitro mutagenesis and selection technique.ThesisItem Open Access STUDIES ON IN VITRO PROPAGATION OF VIRUS INDEXED STRAWBERRY (Fragaria x ananassa Duch.) ITS CONSERVATION AND GENETIC FIDELITY(UHF,NAUNI, 2019-06) THAKUR, KANIKA; GUPTA, RAKESHABSTRACT The present investigation on "Studies on in vitro propagation of virus indexed strawberry (Fragaria x ananassa Duch.) its conservation and genetic fidelity" was carried out at the Department of Biotechnology, Dr Yashwant Singh Parmar University of Horticulture and Forestry, Nauni, Solan (H.P.). The survey done for TRSV in strawberry was confirmed using DAS-ELISA which revealed that no virus was reported from all the three locations (Dhaulakuan, Shimla, Solan). Since the plant material was free from virus an efficient system for micropropagation of strawberry cvs., Selva, Sweet Charlie and Brighton using runner tips was developed. The runner tips was sterilized using 0.2 per cent carbendazim for 10 min + 0.1 per cent HgCl2 for 3 minutes, which were established on MS medium consisting of 0.5 mg/l BA and 0.2 mg/l IBA. The highest multiplication of the established runner tips was obtained when MS medium supplemented with 0.5mg/l BA + 0.20 mg/l IBA + 1.0 mg/l GA3 was used. The maximum rooting of the shoots was obtained on half strength MS medium supplemented with 0.5 mg/l IBA and their maximum hardening was obtained in potting mixture consisting of cocopeat. The various conservation techniques used revealed that the survival of the runner tips at 250C was the maximum but the low temperature condition of 40C was found suitable. The encapsulation of the runner tips was done using 0.5 mg/l BA with maximum regeneration after 4 weeks of storage. Mannitol when used at the rate of 2.5 per cent resulted in 74.19 per cent survival of the encapsulated runner tips after 1 month of storage. Out of the 23 ISSR primers used 17 were informative and revealed similarity between the mother and the micropropagated plants determining the genetic uniformity of the in vitro raised plants.ThesisItem Open Access DEVELOPMENT OF SCAR MARKERS FOR RESISTANCE TO FUNGAL DISEASES FROM CRAB APPLE BIOTYPES(UHF,NAUNI, 2019-05) VIKRANT; MODGIL, MANJUABSTRACT To facilitate the use of marker assisted selection in apple breeding programme, six PCR based specific sequence characterized amplified region (SCAR) markers were developed from crab apple. Seven biotypes of indigenous crab apple biotypes maintained at two field gene banks of Himachal Pradesh state of India were used to identify RAPD markers for resistance to apple scab and woolly aphis. RAPD molecular analysis was evaluated among seven biotypes using 119 primers. Among these, 94 primers generated polymorphism. Four primers OPB-12, OPB-14, OPD-06 and OPA-18 amplified five unique bands of 500bp, 1kbp, 1kbp, 1kbp and 1kbp in M. baccata Shillong, M. baccata Khrot, M. baccata Kinnaur (Dhack), M. baccata Kashmir and M. baccata J&K respectively. These markers were gel purified and cloned into easy cloning TA vectors. Plasmid of the confirmed positive clones after restriction digestion and colony PCR was subjected to sequencing and homology search. Sequences showing homology with the apple scab and woolly aphid resistant genes were used to design pairs of six SCAR primers, which specifically amplified these RAPD fragments in crab apple biotypes as well as M. floribunda and apple rootstock MM106 of known resistance. No amplification was observed in commercially grown varieties of apple, therefore, it is verified that all six SCAR markers were able to distinguish between the resistant and susceptible apple genotypes. Molecular marker assessment with already developed nine SCAR markers in fourteen individual plants of crab apple verified their presence/absence and in accordance with the bioassay results.ThesisItem Open Access IN VITRO SELECTION AND REGENERATION OF SALIX SP. AGAINST CANKER DISEASE(UHF,NAUNI, 2019-05) RANA, SABINA; THAKUR, SANJEEVABSTRACT In the present investigations,in vitro regeneration of commercially importantSalix sp. was carried out and an attempt was made for in vitro selection of this species against Cytospora along with molecular studies. Indirect organogenesis form leaf and inter nodal segment was achieved. Treatment of 12.5% NaOCl for 10 minutes to both explants along with 0.2% bavistin was found to be the best as it gave maximum number of uncontaminated cultures and per cent survival. Callus was induced from leaf and inter nodal explant on MS medium supplemented with 0.5 mg/l NAA with 1.5 mg/l BA and 0.5 mg/l NAA with 0.5 mg/l BA respectively. Highest percentage of callus was obtained from leaf (96.67%) explants followed by inter nodal segment (93.34%). Percent shoot induction from leaf and inter nodal segment derived calli on 0.5 mg/l BA, 0.50 mg/l Kinetin with 0.25 mg/l GA3 and 0.5 mg/l BA with 0.25 mg/l GA3 was 83.33% and 75.00% respectively. Leaf explants was found to be most responsive explants for indirect regeneration. Highest multiplication rate (1:4) was obtained on MS medium fortified with 1.50 mg/l BA with 0.25 mg/l GA3. Shoot number was elevated with the increase in subculturing passages upto third passage thereafter there had been a decline in it. Activated charcoal brings about elongation of shoots. The regenerated shoots were rooted in half strength MS medium containing 500 mg/l activated charcoal. During hardening it was observed that plantlets showed highest survival of 30% in a potting mixture containing sand, soil and cocopeat. To carry out in vitro selection for resistance development, the isolation of canker causing pathogen was done. On the basis of morphological features and BLASTn analysis of ITS region of pathogen, it was identified as Cytospora chrysosperma. The optimum concentration of fungal culture filtrate for selection against canker disease was 30.00 per cent resulting in 12.96 per cent survival of calli. However no shoots were regenerated after selection of calli. For molecular characterization dendrograms were generated to assess the variations among mother plant, control calli and 30 per cent FCF treated calli, using RAPD and ISSR markers which separated them in two clusters where mother plant and control plant always fall in one cluster showing 70 and 79 percent similarity with each other while the selected variants clustered randomly suggesting genetic variation with mother plant, control plant and within the variants.ThesisItem Open Access DEVELOPMENT OF COLLAR ROT RESISTANT SOMACLONAL VARIANTS IN APPLE(UHF,NAUNI, 2019-05) PATIDAR, SHIRISH; MODGIL, MANJUDEVELOPMENT OF COLLAR ROT RESISTANT SOMACLONAL VARIANTS IN APPLE ABSTRACT Apple rootstock MM106 is a commercially productive and semi dwarf tree which is resistant to woolly apple aphid but highly susceptible to collar/crown rot. In the present investigation, an attempt has been made to develop collar rot resistant somaclones using in vitro shoot selection technique. Leaf raised regenerants of MM106 recently selected in vitro on 70-74% fungal culture filtrate (FCF) supplemented MS medium, were exposed to similar concentration of FCF for third continuous selection cycle. It was observed that approx 8187% tolerant regenerants survived on 72% and 70%, while 74% concentration was found inhibitory to all the regenerants. 100% survival was obtained in discontinuous cycle. Multiplication ability was reduced initially but in successive subcultures, FCF tolerant regenerants showed similar multiplication rates as found in axillary bud raised control shoots. Rooting frequency and root system in tolerant regenerants were found better than control shoots. Maximum hardening success of 42.37% was observed in tolerant regenerants. Sixteen FCF tolerant regenerants were screened for resistance in the pathogen inoculated soil. Symptoms like lesion, girdling of stem below the soil level, colour change in roots to reddish brown appeared on control plants, while no such type of symptoms in somaclones. RAPD analysis clearly distinguished the control plants and somaclones. Two unique bands generated by one primer was sequenced which showed leaf rust resistant gene. In conclusion, collar rot resistant plants of apple rootstock MM106 have been developed using in vitro selection of somaclonal variants which may be recommended as cost effective, non- transgenic and promising approach.ThesisItem Open Access STUDIES ON PRODUCTION AND CHARACTERIZATION OF MICROBIAL CHITINASE(UHF,NAUNI, 2019-03) NIRJA; GUPTA, RAKESHABSTRACT The present studies involved isolation, production and characterization of extracellular chitinase from the rhizospheric soil samples of apple orchards of district Shimla and Kinnaur. In total, eighty five bacterial and nine fungal isolates were screened for extracellular chitinase production. Among the thirty bacterial and three fungal isolates producing extracellular chitinase, bacterial isolate SK1.2 and the fungal isolate SNF1.1 were found to be the best. These two isolates were characterized morphologically and molecularly by using 16S and 18S rRNA gene sequencing technique. They were identified asBacillus cereus SK1.2|MF280165| and Alternaria brassicicola SNF1.1|MK024281|. The culture conditions for extracellular chitinase production were standarized for both the bacterial as well as fungal isolates with respect to temperature, pH, incubation time, carbon sources, nitrogen sources and substrate concentration using One Variable at a Time (OVAT) approach. Maximum chitinase production was observed at temperature 30˚C, pH 6.0 and 1% substrate concentration for both the isolates, however incubation time of 72 h for bacterial isolate and 120 h for fungal isolate was found to be optimum. Fructose and dextrose were found to be the best carbon sources for bacterial as well as fungal isolate respectively whereas, ammonium chloride and yeast extract were found to be the best nitrogen sources in culture broth. Two different substrates were tried (Rice bran and termite wings) for production of chitinase under Solid state fermentation (SSF) and maximum production was found with rice bran. The enzyme from bacterial and fungal isolates was partially purified to 3.10 and 3.15 folds respectively using ammonium sulfate precipitation and gel filtration chromatography on Sephadex G-100 column. The partially purified enzymes showed single band on Native-PAGE and single band with molecular weight of 43.7 and 44 kDa on SDS-PAGE for bacterial and fungal isolate respectively. Enzymes were characterized and had a pH optima of 7.0 for bacterial chitinase and 6.6 for fungal chitinase and optimum temperature of 40oC and 35oC for bacterial and fungal chitinase respectively. Out of different metal ions (Cu2+, Mg2+, Ca2+, Fe2+, Mn2+ and Zn2+ ) tested for enzyme activity, Mn2+ and Mg2+ were found to increase enzyme activity. Bacillus cereus SK1.2 chitinase enzyme showed maximum activity at 0.5% of colloidal chitin, after which it falls slightly. The Vmax value was determined by MM equation and confirmed by Lineweaver Burk plot. The value of Vmax and Km was 66.67 μmol/min and 0.13 mg/ml respectively. Bacterial chitinase showed stronger inhibitory activity towards potent phytopathogen in comparsion of control i.e., 66.7% in case of Fusarium oxysporum, 64.6% in case of Rhizoctonia solani and 63% against Colletotrichum gloeosporioides. The fungi Fusarium oxysporum showed significant alterations in its cell wall structure after treatment with purified chitinase as compared to control when observed under TEM and STEM. Similarly, the partially purified chitinase enzyme from Alternaria brassicicola strain SNF1.1 showed strong inhibitory activity against Escherichia coli and Staphylococcus aureus. So, chitinase from these organisms could be effectively employed as potential biocontrol agents against harmful pathogensThesisItem Open Access TARGETING VITAL GENES OF PHYTOPATHOGENIC FUNGUS MARSSONINA CORONARIA USING RNAi APPROACH FOR HOST INDUCED GENE SILENCING IN APPLE(UHF,NAUNI, 2018) CHAUHAN, ARJUN; MODGIL, MANJUABSTRACT Marssonina coronaria (Ellis & J.J. Davis) is economically very important apple pathogen fungus causing severe premature defoliation in apple. All the available commercial cultivars of apple are susceptible to this pathogen. The apple orchard survey revealed that the disease was widespread in three districts of HP. Despite this, nothing much has been studied about the molecular biology of this fungus and its genome remains unannotated. A highly virulent strain of M. coronaria was isolated and characterized. Among different media tested, maximum colony diameter was produced on PPDA whereas, maximum fungal dry weight and conidia were produced in PPDB grown cultures at 25°C. Treatment of conidia with EGTA and anti-calmodulin drug Trifluoperazine dihydrochloride in PPDB resulted in the inhibition of spore germination as compared to the control. Eight RNA extraction methods were evaluated to yield high-quality and melanin-free RNA. Age of fungus culture on solid and liquid medium affected the total RNA yield, and melanin contamination. TRIzol™ Reagent+RNA Clean & Concentrator™-5 and Ambion-PureLink® RNA Mini Kit were found to be the best methods which were applied for RT-PCR, subsequent PCR amplification, and isolation of calmodulin, β-tubulin, HSP-90 co-chaperone Cdc37, SNF2 family domain, C6 zinc finger domain and transcription activator gene sequences from M. coronaria. Semi-qRT-PCR of β-tubulin gene revealed its suitability as reference housekeeping gene for gene expression studies and explored for molecular detection of M. coronaria from infected apple tissues. Two essential genes calmodulin and HSP-90 were opted for dsRNA synthesis using pL4440 vector and development of hairpin-RNAi constructs using pSilent-1 and pCAMBIA-1300 vectors. Treatment of conidia suspension with 10 μL of dsRNA (750 ng/ul) targeting CALM and HSP-90 gene resulted in 11 and 34 colonies, respectively which were significantly less than that of the water control (74). These treated conidia as well as the topical application of dsRNA resulted in reduced symptom appearance on leaf surface of apple varieties. ATMT was standardized with mycelia plugs and fragmented mycelia using pCAMBIA-1304 with parameters viz., Agrobacterium concentration (OD600 - 0.5), AS (250μM), coculture time (48 h), and temperature (24°C) and membrane (Whatman filter paper). pCAMBIA-1304 transformants exhibited strong GUS staining, GFP fluorescence and hygromycin resistance even after five successive culturing, and were PCR positive for the hpt and gus genes. Standardized ATMT was employed to generate silenced mutants with CALM and HSP-90 hairpin constructs and confirmed by PCR for hpt gene. Also, qRT-PCR analysis showed drastic reduction in the transcript level of these genes in mutants as compared to control fungi. The mutants also showed significant reduction in growth, sporulation, mycelia dry weight and pigmentation. In vitro detached apple leaf infection revealed that the spores of the silenced mutants of HSP-90 target gene showed low virulence with delayed symptom appearance whereas, the CALM mutants exhibited loss of virulence. To conclude, we have elucidated the role of calmodulin and HSP-90 co-chaperone Cdc37 genes of M. coronaria by utilizing the RNAi approach. Further, this study will widen the molecular biological knowledge of this fungus and host pathogen interaction as well as HIGS to control apple leaf blotch.ThesisItem Open Access BIOPROSPECTING OF COPPER NANOPARTICLES SYNTHESIZING INDIGENOUS BACTERIA AND POTENTIAL APPLICATIONS(UHF, NAUNI, 2019-01) KASHYAP, PRIYANKA; SHIRKOT, POONAMABSTRACT Nanotechnology is gaining tremendous incentive in the present century due to its capability of modulating metals into their nanosize, which changes the chemical, physical and optical properties of metals.The synthesis of nanostructured materials, especially metallic nanoparticles has gained utmost interest over the few decades owing to their unique properties that make them applicable in different fields of science and technology.Therefore, isolation and identification of bacteria with ability of copper nanoparticles synthesis from natural sources is very important in terms of discovering new industrial products. Keeping in view, copper rich sites located Chambaghat village of district Solan, Sataun village of Sirmour, Seond and Larji village from Kullu district of Himachal Pradesh and Khetri Nagar from Rajasthan were selected as a source for new coppernanoparticles synthesizing bacteria. Therefore, aim of present study was isolation and characterization of copper nanoparticles synthesizing bacteria from these sites for biosynthesis of copper nanoparticles followed by applications of these copper nanoparticles. A total of 154 putative copper nanoparticles synthesizing bacterial isolates were obtained from 22 samples collected from different selected siteswhich were characterized morphologically. Thirty bacterial isolates were selected on the basis of their ability to show maximum copper nanoparticles synthesizing activity and one bacterial isolate exhibiting maximum copper nanoparticles synthesizing activity viz., SCS1.1was selected for further molecular characterization using 16S rrna gene technology. In silico analysis of 16S rrna gene sequence led to identification of this isolate as Stenotrophomonas maltophilia strain SCS1.1. On the basis of maximum copper nanoparticles synthesizing activity Stenotrophomonas maltophilia strain SCS1.1 was selected for in vitro biosynthesis of copper nanoparticles. Maximum copper nanoparticles synthesis was achieved at 30°C, pH 8.0 after 48hrs of incubation with 4.0 mM copper sulphate, 5.0% peptone, 3.0 % beef extract and 4.0% inoculum size. In vitro biosynthesis of copper nanoparticles was carried out using optimum conditions which were characterized using UV-visible spectroscopy, FTIR, XRD, TEM, SEM and DLS. In the present study, ability of copper nanoparticles and bacterial culture suspension of Stenotrophomonas maltophilia strain SCS1.1. was further assessed for degradation of fifteen textile dyes producing significant results, these were also found to inhibit various fungal and bacterial pathogens under invitro and in vivo conditions. It has been observed that 25 ppm of Chlorpyrifos, Profenofos and Imidacloprid were degraded at pH 8 using both copper nanoparticles and Stenotrophomonas maltophilia strain SCS1.1 culture suspension. Effect of different concentrations of copper nanoparticles on plant growth and development of twelve crops were studied and 100 ppm were found best for Solanum lycopersicum and Spinacia oleracea wh