DIFFERENCE BETWEEN NATIVE AND MUTANT RHIZOBIUM ON GROWTH OF VIGNA MUNGO L.

DIFFERENCE BETWEEN NATIVE AND MUTANT RHIZOBIUM ON GROWTH OF VIGNA MUNGO L.

Shijila Rani AS ¹, P. Parthiban ^*1 and V. Mahesh ²

Department of Microbiology ¹, Marudupandiyar College, Vallam, Thanjavur, Tamilnadu, India.

MR Govt. Arts and Science College ², Mannargudi, Tamilnadu, India.

ABSTRACT: The present study, carry out the use of plant growth promoting rhizobacteria, were used to improve the yield in crop. The Rhizobium strains isolated from legumes plant Mimosa pudica L and this strain was mutated by using UV radiation. The plant growth hormones were detected by native as well as mutant Rhizobium strains by paper chromatography techniques. The same strains use in conducted to green house; the compared to two strains of Rhizobium, mutated strain are high growth and yield when compared to normal stain.

Mimosa pudica L, Rhizobium strain, mutated agents, Vigna mungo L

INTRODUCTION: Rhizobia encompass a range of bacterial genera, including Rhizobium, Bradyrhizobium, Sinorhizobium, Mesorhizobium, Allorhizobium and Azorhizobium, which are able to establish a symbiosis with leguminous plants. They elicit the formation of specialized organs, called nodules, to reduce atmospheric nitrogen and make it available to the plant ¹.

In legume-Rhizobium symbioses, variabilities in symbiotic effectiveness which are either due to variations in nitrogen fixing potential of Rhizobium strains ^{2, 3, 4} or due to host genotypic compatibility of DNA based detection technology that plastids and mitochondria of the eukaryotic cell were derived from a consortium of primitive microbes ^{5, 6, 7} are often observed.

The common capacity of such symbiosis to reduce dinitrogen to ammonia and to incorporate this product into the nitrogen metabolic stream of the host plant gives the genus a place of outstanding importance in natural ecosystems and agricultural production.

In genetics, a mutagen is a physical or chemical agent that changes the genetic material, usually DNA, of an organism and thus increases the frequency of mutations above the natural background level. As many mutations can cause cancer, mutagens are therefore also likely to be carcinogens. For example of chemical mutagens are ethidium bromide, Acridine dye, etc and physical mutagens are X-rays, alpha rays, gamma rays, beta rays, cosmic rays, they distort or break DNA duplex and disturb the replication. Ultraviolet rays are non-ionizing radiations and produce thymine dimers. In this paper, they are report the symbiotic behaviour of these mutants in relation to the parent strain.

MATERIALS AND METHODS:

Sample collection: Root nodules were collected from young and healthy seedling plants of Mimosa pudica L, plant from Oruthanadu, Thanjavur (dist), Tamil nadu, India.

Isolation of Rhizobium sp from root nodules: To select, healthy and reddish-pink root nodules were tenderly washed with distilled water and surface sterilized by keeping in 0.1 percent HgCl₂ for 4-5 min and wash with sterile distilled water following by 95 percent ethyl alcohol, repeat washing with sterile distilled water ⁸. The nodules were crushed in a small drop of sterile distilled water to obtain milky suspension. Then, serial dilutions were made and aliquots dilutions were spread on yeast extract mannitol agar (YEMA) medium plates were incubated at 28±1°C for 3-5 days ⁹.

Identification and biochemical test of Rhizobium sp: The isolated colonies are conformed by microscopic observation, biochemical test, ¹⁰ and cultural test were performed. Isolates were identified as per Bergey’s manual of Systematic bacteriology.

Identification of IAA: Biosynthesized auxins were partially characterized by paper chromatography method ¹¹. In which three, grams of culture filtrate were extracted with 50 ml of peroxide-free ether for 2 hours at 5ºC. Extracts of the samples were simultaneously loaded and developed using isopropanol: ammonia:water (10:1:1 v/v/v) as a running solvent. The appearance of the strips under ultraviolet light and spraying with modified Salkowski reagent is indicated.

Effect of native and mutant Rhizobium on growth of Vigna mungo L: The pot trails were accomplished by Department of Microbiology Marudupandiyar College, Vallam, Thanjavur ¹². Each one had conducted triplicates. The treatments follow as:

T1 Control (with out Rhizobium), T2 Rhizobium (R Mutant seed inoculation), T3 Rhizobium (R Native seed inoculation), T4 Rhizobium (R Mutant seed inoculation + foliar spray), T5 Rhizobium (R Native seed inoculation + foliar spray).

After germination, plants were treated with 48 hrs old culture, which were foliar sprayed to the Vigna mungo plants every week for 4 times, starting from 10^th day of germination. The plants were harvested every 15^th day of intervals, after harvesting to analyse agronomical characters such as morphometric and number of root nodules analyse in standard procedure.

RESULT AND DISCUSSION: The isolates of rhizobia from root nodules of legume plants were characterized by opaque and milky white in appearance in YEMA medium. The rhizobial strains were isolated from the nodules of Mung bean ¹³, Pisum sativum ¹⁴ and Cassia alata ¹⁵. The rhizobial strains were identified in microscopic and biochemical test. The result were compared with Bergey’s manual of Determinative Bacteriology 9^th edition and conformed as Rhizobium. Similarly, Singh et al. ¹⁶ also characterized Rhizobium strains on the basis of biochemical tests. The native strain of Rhizobium was induced by UV mutagen treated with different time intervals, (1 to 6 min). Out of six minutes treatment, four minutes were observed in maximum number of colonies (3.0±2.12) compared to other treatments recorded in Table 1. So, further study we have chosen in four minutes mutated isolates.

TABLE 1: NATIVE RHIZOBIUM SPECIES ARE SPONTANEOUS MUTATION BY DIFFERENT TIME

 These Rhizobial strains were use in separation of Auxin and Gibberellin compounds by paper chromatography method by Table 2. Some workers reported by different legume nodulating rhizobial strains preferred different vitamins sources for IAA production reported ^{17, 18}.

 TABLE 2: SEPARATION OF PLANT GROWTH HORMONE BY PAPER CHROMATOGRAPHY TECHNIQUES

The pot trial was conducted to study the response of Vigna mungo to inoculation with Rhizobium and foliar application. The result revealed that the black gram responded well to the inoculation of rhizobium. The plants inoculated with mutant Rhizobium possessed significantly greater plant height, dry weight and nodules formation was monitor, the UV mutant Rhizobium species were high responsible for the plant growth (Table 3, 4 and 5). The current observations of the pot experiment of pea was close to Fischer et al. ¹⁹, who find out the ability of Rhizobium on wheat by development of shoot/ root fresh and dry weights. All the Rhizobium strains improved the root and shoot dry biomass by 100% and 70% respectively. The current conclusion goes to the results ^{20, 21}, who obtained 70% increase in pea root/shoot dry biomass by PGPRs inoculation as contrast to control (uninoculated).

TABLE 3: 15^TH DAY MORPHOMETRIC ANALYSIS OF V.MUNGO PLANT

T1 Control (with out Rhizobium), T2 Rhizobium (R Mutant seed inoculation), T3 Rhizobium (R Native seed inoculation), T4 Rhizobium (R Mutant seed inoculation + foliar spray), T5 Rhizobium (R Native seed inoculation + foliar spray) T6 Combined effect of Native and Rhizobium.

 TABLE 4: 30^TH DAY MORPHOMETRIC ANALYSIS OF V.MUNGO PLANT

T1 Control (with out Rhizobium), T2 Rhizobium (R Mutant seed inoculation), T3 Rhizobium (R Native seed inoculation), T4 Rhizobium (R Mutant seed inoculation + foliar spray), T5 Rhizobium (R Native seed inoculation + foliar spray), T6 Combined effect of Native and Rhizobium.

 TABLE 5: 45^TH DAY MORPHOMETRIC ANALYSIS OF V.MUNGO PLANT

T1 Control (with out Rhizobium), T2 Rhizobium (R Mutant seed inoculation), T3 Rhizobium (R Native seed inoculation), T4 Rhizobium (R Mutant seed inoculation + foliar spray), T5 Rhizobium (R Native seed inoculation + foliar spray) T6 Combined effect of Native and Rhizobium.

CONCLUSION: In the current investigation, the UV mutant Rhizobium sp were treated by V.mungo L., the foliar spray treatment seen highly responsible growth in plant height, leaf area and no. of root nodules, while only treated UV mutant Rhiozobium sp. when compared to control plant.

From this study conclude by some mutation occur in DNA basepair (AGTC), so get high results.

 ACKNOWLEDGEMENT: The authors wish to thankful of Mardupandiyar College, Vallam, Thanjavur, for extending the facilities to carry out the laboratory studies.

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    How to cite this article:

    Rani ASS, Parthiban P and Mahesh V: Difference between native and mutant rhizobium on growth of Vigna mungo L. Int J Pharm Sci Res 2016; 7(9): 3848-51.doi: 10.13040/IJPSR.0975-8232.7(9).3848-51.

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