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CHAPTER 3MATERIALS AND METHODS The presentstudies entitled “Effect of Integrated nutrient managementand Bio-regulater on on growth, yield andquality of broccoli (Brassica oleracea var L.

italica Plenck)” were carried out at the Horticulture Farm,S.K.N. College of Agriculture, Jobner, Jaipur, Rajasthan during Rabiseason 2016-17 and 2017-18. The details ofexperimental site, material used and methodologies employed have been describedas under:3.       Agro-climatic conditions of experimental site3.1       Location of experimental siteTheexperiment was laid out at Horticulture farm, S.K.

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N. College of Agriculture,Jobner, Jaipur (Rajasthan) during Rabiseason 2016-17and 2017-18. Geographically, Jobner is situated 45 km West ofJaipur on 260 5 North latitude, 750 20 Eastlongitude and at an altitude of 427 metres above mean sea level. This regionfalls under agro-climatic zone-IIIA (Semi-Arid Eastern Plain zone) of theRajasthan.3.2       Climate and weatherThe climate of Jobner region is semi-arid characterized by extremes of temperature both in summer and winter,  with low  rainfall  and  moderate relative humidity. The annual average rainfall is 400 to 500 mm, most of which is fall during July to early September; sporadic showers also received inwinters.

The maximum temperature ranges from 30 to 46ºC during May and June, while in December and January, it falls below1ºC and evaporation ranges from 2.5-6.9mm/day. The meteorological data  during   the period   of experimentation recordedat meteorological observatory of the college are presentedinTable 3.

1 and depicted in Fig. 3.1.  3.

3      Soil characteristics of experimentalfield Soil samples taken from experimental field wereanalyzed in the Department of Soil Science and Agricultural Chemistry,S.K.N. college of Agriculture,Jobner to evaluate physico-chemical characteristics of the soil duringexperimentation. The soil samples from 0-15 cm depth were collected fromdifferent locations of the experimental field before application of manures andfertilizers. A representative composite sample was prepared by processing andmixing them together and analysed for physical and chemical properties. Theresults of analysis presented in Table 3.2 showed that the soil of experimentalsite was loamy sand in texture, slightly alkaline in reaction, poor in organiccarbon with low available nitrogen, phosphorus & sulphur and medium inpotassium content.

The underground water used for irrigation is partially salinein nature. The pH and EC of water were 8.1 and 1.9 d/Sm, respectively.3.

1.4  Qualityof irrigation waterThe irrigation water falls under class C4S2,according to USDA Hand Book No. 60. The pH and ECe was found to be 8.1 and 3.2ds m-1, respectively.

Water of Jobner area is saline in nature. Thedetail of irrigation water is given in Table 3.3.3.2       Experimental design and layoutThe experiment was laid out in Split plot design(SPT) with 3 replications.

Randomization of the treatments was done with the help of random number table as advocatedby Fisher, 1950. The plan of layout of experiment is given in Fig. 3.

2. Details of treatments with Notations are given in Table 3.4.    Detailsof experiment are given below: Season and year :       Rabi season, September 2016 to        January 2017 and September               2017 to January 2018  Crop Name                         :     Sprouting Broccoli Variety :      Pusa KTS 1 Experimental Design  :      Split Plot Design Replications :      3 Treatments :      7 x5 = 35  Total number of plots :      105 Spacing  :      45 cm x 45cm Plot size :      2.25 m x 1.80 m No. of plant per plot               :      20 Gross experimental area         :      775.5 m2 Table3.

4       Details of treatments withnotation (A) Fertility levels (main plots) Symbol 1. 100% RDF (100:80:60 kg NPK/ha) F0  2. 75% RDF+ 25% FYM (5 t/ha)                                     F1  3.

50% RDF+ 50% FYM (10 t/ha)                                     F2  4. 100% FYM (20 t/ha)                                                                        F3  5. 75%RDF+ 25% VC (1.

75 t/ha)                                                                            F4  6. 50% RDF + 50% VC (3.5 t/ha)                                                                                                     F5  7. 100% VC (7 t/ha)                                                                                                                             F6  (B) Bio-regulators (sub-plots) Symbol 1. Control (water spray) B0  2.

Brassinoids (5 ppm) B1  3. Brassinoids (10 ppm) B2  4. Salicylic acid (100 ppm) B3  5. Salicylic acid (150 ppm) B4   Thequantity of mulching material as well as biofertilizers are as under used. S.No.

Mulching material/ biofertizer         Quantity/ thickness 1 Mustard straw                                5t/ha 2 Saw dust                                        10t/ha 3 Black polythene sheet 200 gauge thickness 4 Phosphate Solubilizing Bacteria(PSB) 2kg/ha 5 Azotobacter 2kg/ha  Table 3.5       Details of the treatments alongwithcombinations. Treatment Combination Treatment Combination T1 M0B0 T9 M2B0 T2 M0B1 T10 M2B1 T3 M0B2 T11 M2B2 T4 M0B3 T12 M2B3 T5 M1B0 T13 M3B0 T6 M1B1 T14 M3B1 T7 M1B2 T15 M3B2 T8 M1B3 T16 M3B3 3.3       Characteristicsof variety Pusa KTS -1            Itis a medium-tall (65-70 cm) variety. Foliage is waxy and dark green withslightly wavy margins. Heads are solid green with small beads slightly raisedat the centre.

The main head size and weight are about 6.0-15. cm and 350-450 grespectively. It matures in 90-105 days after transplanting under temperateclimate, while 5-10 days earlier in the tropical plains.

3.3     Raising of the experimental crop3.3.1   Preparation of nursery beds and rising of the seedlingTwo raised nursery beds of 3 × 1 × 0.15 m size wereprepared by mixing well rotten farm yard manure in soil at the rate of 15 kgper square metre. Seeds were treated with 0.02 per cent thiram to cheek theproblem of damping off in the seedling Seeds were sown on 5thSeptember, 2016 in shallow furrows 5-6 cm apart by dropping the seeds at 1-2 cmdepth.

A thin layer of powdered leaf mould was applied to cover the seed.Regular watering, hoeing, weeding, plant protection measures, etc. were done time to time. The seedlings were ready for transplantingwithin six weeks after sowing of seeds in the nursary beds.3.

3.2   Field preparationThe experimental field was thoroughly ploughed and cross ploughedwith the help of mould board plough and cross-harrowing was done with tractor,followed by planking and leveling to bring the field to a good tilth. Beds of1.

8 × 1.8 m size and paths and channels were also prepared according to thelayout plan of the experiment.3.3.

3   Application of treatments3.3.3.1  Bio-fertilizers:Thesprouting broccoli seedlings were treated with PSB and Azotobacter culture alone as well as in combination of both as pertreatment plan, using standard methods. Suspension of half kilogrambio-fertilizer in seven litres of water was prepared for treatment of seedlingsto transplant in 155.52 m2.

Prior to required quantity of culturewas mixed in cold jaggery water and the seedling of sprouting broccoli weredipped in solution for 10 minutes before transplanting.3.3.3.2  Mulching:Mulchingwas done prior to transplanting of over seedling. The  black polythene sheet of 200 gauge thicknesswas spread beds and mustard straw and saw dust were put into bed size 1.

8 x 1.8m and the holes of 1 x 1 inch were made on polythene sheets as per the plantand row to row distance similiarly.Mustard straw and saw dust @ 5t/ha & 10t/ha were spread of above 1 cmthickness as mulch on top soil of the beds and thereafter, transplanting of theseedling was done at appropriate distance in beds.3.3.

4 Transplanting ofseedlings Aboutsix weeks old seedlings of sprouting broccoli were transplanted in the field on21th October, 2016 and the average height of the seedlings was about8-10 cm. The distance between row to row and plant to plant was kept 45 cm x 45cm. Thus, 16 plants were accommodated in each plot. The transplanting was donein evening hours followed by light irrigation3.3.5   IrrigationsThecrop was irrigated immediately after transplanting and then at an intervalof 2-3 days upto the establishment of seedlings.After this, the crop was irrigated at regular intervals of 8to 10days.

3.3.6   Gap filling Gapfilling was done in place of unsuccessful or dead seedlings in the initialperiod of the crop to maintain the plant population in each plot uniformly.3.

3.7   Weeding and hoeingTo maintain the proper plant stand, firstweeding and hoeing was done at 20 days after transplanting and later on hoeingand weeding  at 15days interval and twowere performed.3.3.8   Plant protection Inorder to protect the crop from insect, pest and diseases standard methods ofplant protection were followed. To check the attack of aphid, butterfly,looper, prophylactic measures were taken i.

e.sprouting broccoli plant were sprayed with Malathion 50 EC at 0.05 per centafter transplanting to protect sprouting broccoli from insect and pest attack.3.3.8   Harvesting of curdsCurdswere harvested before opening of bud when the bud clusters were compact.Harvesting was done with the help of sharp sickle and observations of taggedplants were recorded. The process of harvesting was startedfrom first week of January, 2017 to last week of February, 2017.

3.4       Characters studied and observationrecordedIn order to evaluate the “Effect of Mulching and Bio-fertilizers onGrowth, Yield and Quality of Sprouting Broccoli (Brassica oleraceae var.italica L.)” five plantswere randomly selected and tagged from each plot and observations were recorded. The methodology of individual aspect isbriefly described in the following paragraphs.3.4.1  Growthparameters 3.

4.1.1  Plantheight (cm)3.2 Experimentaldetails3.

2.1 CultivarCultivar ‘Green Head’ of broccoliwas chosen for the prsesent study. The cultivar has been recommended forcultivation in different agro-climatic zones of Uttarakhand. Head of thecultivar is dark green, medium in size and rich in vitamin A.

The average yieldof cultivar ranges from 100-175 q/ha.3.2.2 TreatmentsThe presentstudy comprised of 10 treatments, which consisted of sole application oforganic sources (vermicompost, neem cake, biovita granules and farmyardmanure), biofertilizers (Azotobacter andPhosphate Solubilizing Bacteria) and their combinations as detailed in Table 3.2.

Experimentaldetails:- The details of theexperiment to be conducted in 2015 as underExperimentDesign                 –  Randomized Complete Block Design (RCBD)Treatment                               –   10Replication                             –   03Spacing                                  –   45 cm x 45 cmPlotsize                                 –   1.8m x1.35m (2.43m2)Numberof plant per plot       –   12Table 3.

2. Detail oftreatmentsTreatment code                   Treatment Detail1.     T1             –                Farmyard manure  (FYM) @20t/ha2.     T2             –                Vermicompost @5t/ha3.

     T3             –                Neem cake @ 2t/ha4.     T4             –                Biovita granules @50 kg/ha5.     T5             –                Biofertilizers (Azotobacter+PSB) each @5kg/ ha 6.     T6             –                Farmyard manure +  Biofertilizers 7.     T7             –                Vermicompost + Biofertilizers8.

     T8             –                Neem cake + Biofertilizers9.     T9             –                 Biovita granules +Biofertilizers10.  T10           –                 Control    N  Allotment of treatments in experiment fieldunder RBD Replication-1 Replication-2 Replication-3 T-5 T-2 T-9 T-2 T-3 T-10 T-3 T-7 T-6 T-4 T-5 T-7 T-10 T-1 T-8 T-7 T-9 T-1 T-6 T-10 T-3 T-8 T-6 T-4 T-9 T-8 T-2 T-1 T-4 T-5  3.2.

3 Procurementof seed            Theseed of the cultivar “Green Head” was procured from Department of VegetableScience College of Horticulture VCSGUttarakhand University Of Horticulture And Forestry, Bharsar, Pauri Garhwal,Uttarakhand,India, 246123 3.2.4Nursery raisingThe seed of thecultivar was sown in the well prepared nursery bed in lines and covered with alayer of soil and farmyard manure mixture. Further, these beds were coveredwith a thin layer of dried grass as mulch and watered with the help of rosecane. Regular watering was done in these beds to maintain moisture at fieldcapacity for the proper growth and development of seedlings. After germinationof the seedlings, dry grass was removed to expose the seedlings to sunlight forbetter growth.

Nursery was kept free from weeds. Timely plant protectionmeasures were also followed to prevent nursery from damping off and otherdiseases.3.

2.5Field PreparationThe experimentalfield was thoroughly ploughed by tractor 15 days prior to date oftransplanting. Stones, pebbles and residues of previous crop were removed.Experimental field was levelled properly and sufficient provision for drainagewas made. There after, the layout of the experiment was done, plots wereprepared and treatments were allocated according to the layout plan. 3.3 Methodof different treatment application3.

3.1Application of organic manuresThe entirecalculated dose of vermicompost, neem cake, biovita granules and farmyardmanure as per treatment combination were applied in the individual specifiedplots before transplanting of the seedlings by broadcasting method and wasthoroughly mixed up well with the soil.3.3.2Application of biofertilizersThebiofertilizers (Azotobacter and PSB)were applied through seedlings dip method. A solution of Azotobacter and PSB was prepared by dissolving 200 grams of eachbiofertilizers in 5 litres of water and seedlings were dipped in this solutionfor 30 minutes before transplanting.

After dipping, seedlings were immediatelytransplanted in the field.3.3.3Harvesting The greencompact heads and axillary sprouts were harvested at full maturity stage withthe help of sharp knife. The harvesting was carried out in stages as per thematurity of central head and secondary heads. The first harvesting of heads wasstarted from third week of November 2015.

3.4Observations recordedThe following observations wererecorded:3.4.

1Days taken to 50 per cent headingThis observationwas recorded by visiting the experimental field daily and numbers of days werecounted right from the date of transplanting of seedlings to the date, whenhead were matured in 50 per cent of the plants per plot.3.4.2Plant height at maturity (cm)Plant height wasmeasured from the ground level to the top of the longest leaf at the time ofharvesting with the help of measuring scale. The height of randomly selectedten plants was measured and the average value was expressed in centimetres.

3.4.3Number of leaves per plantAll the fullygrown leaves were counted except for the leaves attached to the heads. Thenumber of leaves were counted randomly in selected five plants and averaged toget number of leaves per plant.3.4.4 Head size(cm2)Polar andequatorial diameter of each head was measured in centimetres on five randomlyselected plants and accordingly multiplied to obtain the head size and meanvalue was worked out, which was expressed in cm2.3.

4.5Average head weight (g)Randomly fiveheads from different plants were selected from each plot, their weight wasrecorded and average value was expressed in grams.3.4.6Number of spears /plantThe number ofsecondary marketable heads were counted on five randomly selected plants fromeach plot and mean value was worked out.3.

4.7Average spears weight (g)The weight ofsecondary heads was recorded on randomly selected five plants from each plotand mean value was worked out and was expressed in grams.3.4.8Average plant yield (g)            Total weight ofterminal head and spear weight was divided by number of plants to obtainaverage plant yield.3.

4.9Yield quintal per hectare (q/ha)The yield datapertaining to central head and secondary heads was recorded on all the plants (q/ha)and accordingly yield per hectare was calculated and expressed in q/ha.3.4.10Shelf life (days) Shelf life offruits was estimated by keeping the fruits at ambient room temperatureconditions till they shrunk and become unfit for consumption.3.

4.11Total Soluble Solids (°B)The total soluble solids content infruits was determined by Erma Hand Refractometer (0 to 32°Brix). Therefractometer was calibrated with distilled water before use and then a fewdrops of fruit juice were placed on the prism and the reading was recorded.

Atemperature correction was applied when it was above or below 20°C (AOAC, 1970).The results were expressed in °Brix.3.4.12  Ascorbic acid (mg/100g)Vitamin C is biochemically known as ascorbicacid, which was estimated by titrimeter method as suggested by Ranganna (1995).

Aliquots were prepared by grinding of well mixed head sample along withmetaphosphoric acid solution and titrated against 2-6, dichlorophenolindophenol dye to pink end point. The vitamin C content was calculated by usingformula as given below: Vitamin C(mg/100g) = x 100 Dye factor                     =  3.5Cost of cultivation Cost ofcultivation was calculated on the basis of prevailing local charges fordifferent inputs like labourer, implements, seeds, fertilizers and otherchemicals, used in cultivation of crops under different treatments.

3.5.1       Grossreturns The sale rate ofbroccoli heads and / spears, leaves and plant stalk yield of Broccoli wereconverted into gross return (Rs./ha) on the basis of prevailing local marketprices of produce ( broccoli heads and /spears/ leaves).3.5.2  Net returnsThe net returnof each treatment was calculated by deducting the cost of cultivation from thegross return of individual treatment.

3.5.3  Benefit: Cost ratioBenefit– costratio was calculated as follows:Benefit: cost ratio =  3.

6  Statistical analysisThe statisticalanalysis was carried out for each observed character under the study usingMS-Excel, OPSTATE. The mean values of data were subjected to analysis ofvariance and ANOVA was set as per Gomez and Gomez (1984) for RandomizedComplete Block Design. For estimation of different statistical parameters,following procedure and formulae were adopted:                                          Analysis of variance Source of variance Degree of freedom Sum of squares Mean sum of squares Variance ratio (V.R.) Replication (r) r-1 Sr Sr/(r-1)           =   Mr Mr/Me Treatments (t) k-1 Sk Sg/(t-1)          =   Mt Mt/Me Error (e) (r-1) (k-1) Se Se/(r-1) (g-1)  =   Me   Where,r            =    Number of replicationsk           =     Number of treatmentsSr          =     Sum of squares due to replicationsSk         =     Sum of squares due to treatmentsSe         =     Sum of squares due to errorMr        =     Mean sum of squares due to replicationsMk        =     Mean sum of squares due to treatmentsMe        =    Mean sum of squares due to errorThecalculated F-value was compared with tabulated F-value.

When F-test was foundsignificant, critical difference was calculated to find out the superiority ofone entry over the others.Thestandard error and critical differences were calculated as follows:SE(m) ±           =          SE(d) ±           =          CD0.05               =          S.E.(d) x t (0.05) (r-1) (k-1) df Where,SE(m) ±           =         Standard error of meanSE(d) ±           =          Standard error of differenceCD0.05               =          Criticaldifference at 5 per cent level of significance

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