Introduction

Methylotrophs a potent bioinoculant to improve crop production. These organism oxidize environmental methane as a sole source of carbon (Quayle, 1961) for their growth either aerobically (Hallem etal, 2004) or anaerobic environment. Green and Bousefield stated that these organisms are predominantly present in paddy field soil and phyllosphere because of high methane emission in paddy field. (New Scientist magazine, 1989). The methane source in the paddy field supports the growth of methylotrophs (Charles etal, 2005). Methylotrophs can be easily cultured in laboratory using some specialized media. Methylotrophs are agriculturally important organisms by producing growth hormone vitamins and pigments (Corpe, 1985; Mahadevan, 1985; Holland and Polacco, 1994; Rohmer, 1999; Koeing, 2002). Various invitro culturing of plants have been introduced so for to identify Agrobacterial characteristics of plant seeds using bioinoculant (Smith, 1992).  New approach in invitro growth studies has been introduced as culture cotton method which are not been used previously.

Materials and Methods

Healthy Rice, maize and Groundnut seeds were selected and surface sterilized. Sterile Pertiplates were taken and substratum for seeds (Cotton) was carefully kept in plates. Pure culture of Methylobacterium was added in the medium and the seeds were sown separately in each petriplate. Plates were kept undisturbed for few days to check Agrobacterial characters of the seed.

Result and Discussion     

Plants should be subjected to analysis of growth characteristics in invitro before going hardening. Various methods can be followed for Agrobacterial character analysis in plants in invitro. Current invitro methods like Rhizobium inoculation in pot culture and foliar spray have certain disadvantages. A novel and simple approach was introduced to analyse thee growth characteristics like seed germination, shoot and root elongation, leaf area meter in invitro condition. By getting the rough analysis once can come to a conclusion about the field growth of the particular plant. Log phase broth culture was taken for as inoculants for its high metabolic activity in that stage. Pure culture of Methylobacterium was taken, it was poured in sterile cotton as a substratum for growing seeds. To avoid other group of microorganism contamination, which imparts in plant growth.  Medium was added during the growth period of seed if the medium is exhausted. This is because the depleted medium should not stop the growth of the organism. Growth was monitored for various seeds and it was tabulated. ( Table-1and Plate-1).

Table 1: Agrobacterial characters of plants using culture cotton inoculation

Plate 1: Growth of seeds using culture cotton method.   Click here to View figure

References

1. Charles M. Maliti, Dominick V. Basile, and William A. Corpe 2005 Effects of Methylobacterium spp. strains on rice (Oryza sativa) L.callus induction plantlet generation and seedlings growth invitro The Journal of the Torrey Botanical Society.  132(2):355-367.
2. Corpe,W.A.and S.Rheem.1989.Ecology of the Methylotrophic bacteria on living leaf surfaces FEMS Microbiology letters.62(4): 243-249.
3. Hallam ,S.J., Putnam, N., Preston, C.M., Detter ,J.C., Rokhsar, D., Richardson, P.M., DeLong. E.F .2004 Reverse methanogenesis: testing the hypothesis with environmental genomics. Science. 305:1457-1462.
4. Holland, M.A., and J.C.Polacco.1994.PPFMs and other convert contaminants: Is there more to plant physiology than just plant. Ann.Rev.Plant Physiol.Plant Mol.Biol. 45: 197-207.
5. Koenig,R.L., R.O.Morris and J.C.Polacco.2002.tRNA is the source of low level trans zeatin production in Methylobacteium spp. J.Bacteriol 184:1832-1842.
6. Mahadevan ,J.1985. Ecology of Methylotrophic bacteria. Current Microbiology.11: 6-9.
7. Quayle, J. R. 1961. Metabolism of C, compounds in autotrophic and heterotrophic microorganisms. Annu. Rev. Microbiol. 15:119-152.
8. Rohmer,M. 1999.The discovery of a mevalonate- independent pathway for isoprenoid biosynthesis in algae, and higher plants.Nat.Prod.Rep.16:565-574.
9. Smith, R. S. 1992. Legume inoculant formulation and application. Can J. Microbiol. 25:739-745.