PROJECT PPP

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SELECTION OF SALT TOLERANT RHIZOBIUM STRAIN Submitted By: TANMOY BANDYOPADHAYAY Ramakrishna Mission Ashrama, Narendrapur, Kolkata, West Bengal ,700103 Ramakrishna Mission Vivekananda University ABBT, 2 nd SEMESTER Supervisor: Dr. S.C. POI Professor, Bihan Chandra Krishi Vishavidyalaya (B.C.K.V )

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INTRODUCTION It is known that legumes enrich the soil by contributing nitrogen through symbiotic nitrogen fixation by Rhizobium through centuries. However, scientific demonstration of value of legumes in contributing nitrogen nutrition of plants was only done in 19 th Century. This was established by the facts that nodules on legume roots are responsible for fixing atmospheric nitrogen through bacterium Rhizobium Salinity of the soil is known to affect both plant growth as well as the occurrence , growth & survival of RHIZOBIUM. However , this micro symbiosis is often affected by lower PH & salinity, keeping the above in view , the present study were undertaken to find out the feasibility of cultivating.

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one pulse crop , Greengram ( Vigna radiata ) were considered during the presence study and there Bradyrhizobium strain were selected for inoculation and study salt tolerant purposes. The plant inoculation experiment were conducted in pot condition. The inoculated seed were pelleted with Gypsum (CaSO 4 ,2H 2 O) before sowing. Besides there were control pot which received neither of the above treatment. The experiment were in R and D . The population of rhizobia in the rhizosphere were studied by pellating method. The experiment was conducted in the pod at Ramakrishna Mission Ashrama ,NARENDRAPUR .. CONTINUE…

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OBJECTIVE Selection of salt tolerant Rhizobium strain specific for greengram for enhancing nodulation in saline zone. To study the efficiency of the salt tolerant Rhizobium strain in nodulation in greengram in pot. To observe the efficacy of pelleting in increasing no of Rhizobium and in germination of seed.

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LITERATURE REVIEW Dadarwal & sen in 1974 reported that The competitive ability of a strain of Rhizobium from mung ( Vigna radiate ) is dependent on the genotype of the host. Bharadwaj (1972) reported that Rhizobium strains from ( Sesbania cannabina ) growing in a saline alkaline environment are more tolerant of these conditions than strains for a normal soil. Bharadwaj (1974) reported that In a highly saline- soil of PH 10.3 , nodule scoring has been delayed until 60 days. Poi etal. (1983) , they reported that appropriate methodology has to be worked out so that the farmers may peiiet the seeds with inoculum in order to grow fodder legume in high acedic soils of North Bengal.

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CONTINUE… Jordan & Aiien in 1974 they reported that species differentiation of Rhizobium is made on the basis of its ablity to nodulate a group related legumes but not others. Kabi & Bhaduri ( 1978) , they reported that he number of nodules also varies in respect of ploidy level of the host. Wilson (1931) reported that Rhizobia are sensitive to salinity with NaCl being less harmful sodium suiphate , potassium sulphate or potassium chloride.

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MATERIALS S1 - strain of Bradyrhizobium of Greengram,selected from BASANTI. S2 - strain selected from GOSABA . and S3 - strain selected from KAIKHALI saline zone.

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The seed of saline zone cultivar of green gram were collected from Sundarban Area through selection (Basis of yield & nodulation). CONTINUE…

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METHODOLOGY YEMA broth preparation Autoclaveing Innoculation of RHIZOBIUM in Broth

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Shaking for 96 hr s CONTINUE… Seed Treatment

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CONTINUE… Some Seeds are pelleting witw Gypsum Different Replication

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CONTINUE… Total 32 pods are taken, among those pods are 8 pods are marked for control and among them 4 pods marked for pelleted practice. Another 24 pods are used for saline trial replication. 24 pods are divided into 3 trial in saline soil for another replication, markly 2%, 1.5% , 0.5%. Among those each (8 pods for each replication) replication again 4 pods marked for pelleting experiment. Irrigation done after 2-3 days interval.

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FINDING AND DATA ANALYSIS : component Amount calcium hydrogen phosphate 0.215 mg , MnSO 4 0.1 gm FeCl 2 0.033 gm CaCl 2 0.053 gm KNO 3 0.06 gm Manitol 10 gm Thamine HCl 100 µgm , Biotine 250 µgm Distille Water 1 lit Determination of Toxic Level Of SODIUM & CHLORIDE IONS in Broth Culture: Burgersense medium :

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( CaCl 2 was added to give in cl + or cl - concentration 0 to 1% ). In addition to the .004% of chloride ion ( cl -) already present in the basal medium flask ( 250 ml) each containing 60 ml of medium were inoculating each with 2 drops of a 3 day culture of Rhizobia grown in basal medium & incubated at 28 0 on a rotary shaker.

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STAIN SALT EXPRESSED AS % Cl- Viable cell / ml after ( week) 0 1 4 12 S1 0.14 8 9 5 5 0.26 8 6 2 1 0.59 8 4 - - 0.88 8 4 - - S2 0.14 14 16 12 10 0.26 15 10 5 2 0.59 14 10 - - 0.88 14 10 - - S3 0.14 10 12 9 8 0.26 10 10 8 4 0.59 10 8 6 2 0.88 10 8 5 2 Table -1. THE EFFECT OF SALT CONCONTRATION & THE SURVIVAL OF BRADY-RHIZOBIUM IN GREENGRAM .

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TREATMENT NO OF RHIZOBIUM IN SEED AT THE TIME OF SOWING CROP NO OF RHIZOBIUM AFTER SET OF FLOWERING INOCULATED CONTROL 48×10 9 15×10 3 GYPSUM PELLETED INOCULATED SEED 48×10 9 73×10 3 Tab-2.POPULATION STATUS OF RHIZOBIUM IN DIFFERENT TREATMENT UNDER SALINE SOIL. It was already reported by Bharadwaj(1972) that Gypsum show reduce salinity and mark Stimulatory effect on the growth and multiplication of the indigenous Rhizobium strain in highly saline soil .

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TREATMENT SURVIVAL RATE OF HOST % NODULE NO Dr y Wt OF PLANT (gm) INOCULATED CONTROL 40 2 2.36 GYPSUM PELLETING 70 15 5.76 C-D at 5% 10 1.5 1.85 Tab-3,. EFFECT OF SEED PELLETING WITH GYPSUM TREATMENT ON THE SURVIVAL , SYMBIOTIC PERFORMANCE OF BLACKGRAM IN SALINE SOIL. In the present study it was found that application of Gypsum as Pelleting agent of inoculated seed improve the survival rate of broth micro and macro symbion and also increased the rate of nodulation and yield of green gram in the saline soil . The application of gypsum was effective as Pelleing agent.

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Nod No/plant Nod Fresh Wt/plant(gm) Plant Fresh wt/plant (gm) Dry plant wt /plant(gm) Pod No/plant Seed wt /plant((gm) Control 12 0.25 15.0 1.56 10 6 Control(P) 16 0.28 18.9 1.95 12 8 2% 0 0 2.3 0.25 2 1 2%(P) 4 0.05 4.5 0.46 4 2 1.5% 5 0.09 5.9 0.62 4 2 1.5%(P) 10 0.16 6.75 0.71 7 6 0.5% 10 0.21 11.5 1.22 15 10 0.5%(P) 18 0.32 25 2.61 18 12 CD at 5% 2.0 0.01 1.25 0.033 0.15 0.025 Tab-4. Table showing the effect of pelleting at different saline soil.

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Fig – shoot Growth after 45 days in control method Fig - shoot Growth after 45 days in control pelleting method Fig - shoot Growth after 45 days in 0. 5% saline soil ( where pelleting done).

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Conclusion : 1. work confirm that present investigation of inoculated seed Pelleting with Gypsum actually increase the nodulation, N 2 - fixation and yield of Greengram in saline soil. 2. But even then the yield remain still inferior to that obtain from the normal soil. It was also found that survival rate of the host is very poor in saline soil. 3. Hence salt tolerant host line should have to be raised for further improvement of the crop in saline soil.

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Implication : The result indicated that the yield of Greengram can be improved in saline soil by applying appropriate salt tolerance strain and the inoculated seed pelleted with Gypsum . 2. So , there is a scope to improve the pulse production in the state by applying the pelleting technology. Hence salt tolerant host line should have to be raised for further improvement of the crop in saline soil.

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Recommendation : The host strains of the present study to be evaluated with other strains having better efficiency in the saline zone. Role of other pelleting ( such as use of rice boiling water ) should be evaluated. N 2 fixing ability of the strain should be evaluated.

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ACKNOWLEDGEMENT

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THANK YOU