In this study, we compared the levels of methylotrophic bacterial community diversity in the leaf samples of 19 rice cultivars collected from three regions of Korea. Nineteen pink pigmented isolates showing characteristic growth on methanol were obtained. Physiological and biochemical characters of each isolate were examined according to methods described in Bergey's Manual of Systematic Bacteriology. When phylotypes were defined by performing numerical analysis of 37 characteristics, four distinct clusters were formed. The two reference strains, Methylobacterium extorquens AM1 and Methylobacterium fujisawaense KACC10744 were found to group under cluster IV and cluster III respectively. Cluster I diverged on the basis of nitrate reduction and four isolates showed tolerance upto 0.5 M NaCl concentrations. Two strains in cluster I and III were found to possess methane utilizing properties. Most of the isolates in all the four clusters utilized monosaccharides, disaccharide and polyols as carbon source. When the isolates were subjected for indole-3-acetic acid (IAA) analysis in the presence of L-tryptophan, only 8 isolates exhibited IAA production. In addition, the nitrogen source in the medium was found to influence the IAA production. Addition of $(NH_4)_2SO_4$ in the medium led to a 2 to 30 fold increase in the indole synthesis. However, $KNO_3$, $NH_4NO_3$ and $NH_4Cl$ substitution did not significantly stimulate the synthesis of IAA in the growth medium. Result of gnotobiotic root elongation assay significantly increased roots and shoots lengths, and number of lateral roots, which is mediated by IAA production in the culture medium. The rice seedlings primary roots from seeds treated with methylotrophic isolates were on average 27 to 56% longer than the roots from seeds treated with the uninoculated seeds. In addition, application of different high concentrations of authentic IAA ($400g\;mL^{-1}$) to roots of rice seedlings inhibited root growth. However, the IAA concentration from 10 to $200g\;mL^{-1}$, IAA promoted root growth of rice seedlings. These results suggest that bacterial IAA plays a major role in the development of the host plant root system.
In order to interpret the effect of IAA on the phosphate metabolism and biosynthesis of organic compounds, Saccharomyces uvarum were cultured in the media treated with various concnetration of IAA $(10^{-3}M,\;10^{-5}M,\;10^{-7}M)$. Sampling at the beginning and intervals of culture, yeast cells fractionated were traced the contents of inorganic phosphate and organic compounds of various fractions. 1. Growth of Saccharomyces uvarum were enhanced by IAA $(10^{-3}M,\;10^{-5}M)$ and phosphate contents in DNA and RNA fractions treated with IAA were accelerated 2.3 times and 2 times in comparison with those of control. 2. Amounts of poly-P"A" and poly-p"B" were increased but poly-P"C" decreased during the culture. Therefore, it is considered that poly-P"C" play on most important role as a phosphate pool. 3. It is suggested that because phosphate contents in DNA, protein and lipid fractions increased, inorganic phosphates required phosphates required RNA were transferred from phosphates in cytoplasm, because these increased slowly during the culture. 4. Alkali-labile protein were accelerated by IAA and alkali stable protein only were inhibiction were enhanced by IAA while, ethanol : ether soluble fraction was induced by $10^{-7}M$ IAA in comparison with those control.X> IAA in comparison with those control.
Journal of the Korean Society of Food Science and Nutrition
/
v.17
no.2
/
pp.115-124
/
1988
This study was carried out to realize the effect of gibberllic acid$(GA_3)$, 1-naphthaleneacetic acid(NAA) and indole-3-acetic acid(IAA) on the biosynthesis of vitamin C. The relation between carbohydrate metabolism and vitamin C production in soybean sprouts was also investigated. Growth, vitamin C content, protein, galactonolactone dehydrogenase(GLD), ribulose diphosphate carboxylase(RuDpCO) and RNA level in the plastid and cytoplasm were determined. The effects of protein and respiratory inhibitors on the growth and vitamin C production were also examined. The most favourable growth of soybean sprouts was observed at the level of NAA $10^{-8}M,\;IAA10^{-6}M\;and\;GA_3\;10^{-5}M$ in the single treatment, respectively, and also favourable at levels of $GA_3\;10^{-5}M+NAA\;10^{-9}M\;and\;GA_3\;10^{-5}M+IAA\;10^{-9}M$ in the case of mixed treatment. The excellent growth was observed at the level IAA $10^{-6}M$ among all the single and mixed treatments. When the soybean sprouts were treated with NAA $10^{-8}M,\;IAA\;10^{-6}M\;GA_3\;10^{-5}M,\;GA_3\;10^{-8}M+IAA\;10^{-6}M,\;and\;GA_3\;10^{-5}M+IAA\;10^{-9}M$, the maximum growth rate was observed at the level of IAA $10^{-6}M$ and the conten of vitamin C was 24.26mg% which was 1.6 times higher than that of the control. RuDpCO was inhibited by the chloramphenicol at the concentration that did not inhibit the growth but the activities of NADP-GDH, GLD and vitamin C content were not affected. These results showed that the biosynthesis of viamin C had nothing to do with the activity of chloroplastic RNA but with cytoplasm. The highest vitamin C content was found at the the level of IAA $10^{-6}M$, where the GLD activity increased up 1.8 times of the control. The concentration of IAA $10^{-6}M$ promoted the biosynthesis of RNa and protein both in chloroplast and cytoplasm, especially in the cytoplasm. Thus it suggeted that IAA affected vitamin C biosynthesis by regulating RNA level in the cytoplasm. 2,4-Dinitrophenol as an uncoupler of oxidative phosphorylation did not inhibit the vitamin C biosynthesis, however, all of the respiratory inhibitors severely inhibited the growth and vitamin C biosynthesis.
Peach (Prunus persica L.) is a model species for stone fruit studies within the Rosaceae family. Auxin plays an important role in the development of peach fruit. To reveal the distribution of auxin in the tissues of peach fruit, immunohistochemical localization of IAA was carried out in the seed, mesocarp, and endocarp in developing peach fruit using an anti-indole-3-acetic acid (anti-IAA) monoclonal antibody. A strong IAA signal was observed throughout the outer and inner integument during peach fruit development, and the distribution was zonal. The IAA signal was mainly focused in mucilage layers in the outer integument. The outer integument may function to produce or store IAA in the seed; a strong IAA signal was detected in the cells around the vascular tissue, whereas a weak IAA signal was located in the vascular tissues. In the mesocarp, the cells around the vascular bundle tissue gave rise to an IAA signal that increased in the late phase of fruit growth, which coincided with a significant increase in fruit growth. The distribution of IAA, however, was changed when fruit was treated with auxin transport inhibitors NPA (1-N-naphthylphthalamic acid) or TIBA (2, 3, 5-triiodobenzoic acid); in mesocarp tissues, an IAA signal was detected mainly in vessels of the treated fruit. During the critical period of endocarp lignification, the vessel lignification process was negatively correlated with IAA signal. The present results confirmed that the distribution of IAA was different in various tissues of peach fruit according to the developmental stage. This research provides cytological data for further study of the regulatory mechanism of auxin in peach fruit.
Bacteria from the Methylobacterium genus, called pink-pigmented facultative methylotrophic bacteria (PPFMs), are common inhabitants of plants, potentially dominating the phyllosphere population, and are also encountered in the rhizosphere, seeds, and other parts of plants, being versatile in nature. The consistent success of the Methylobacterium plant association relies on methylotrophy, the ability to utilize the one-carbon compound methanol emitted by plants. However, the efficiency of Methylobacterium in plant growth promotion could be better exploited and thus has attracted increasing interest in recent years. Accordingly, the present study investigated the inoculation effects of Methylobacterium sp. strains CBMB20 and CBMB 110 on seed imbibition to tomato and red pepper on the growth and accumulation of phytohormone levels under gnotobiotic conditions. Seeds treated with the Methylobacterium strains showed a significant increase in root length when compared with either the uninoculated control or Methylobacterium extorquens $miaA^-$ knockout mutanttreated seeds. Extracts of the plant samples were used for indole-3-acetic acid (IAA), trans-zeatin riboside (t-ZR), and dihydrozeatin riboside (DHZR) assays by immunoanalysis. The treatment with Methylobacterium sp. CBMB20 or CBMB 110 produced significant increases in the accumulation of IAA and the cytokinins t-ZR and DHZR in the red pepper extracts, whereas no IAA was detected in the tomato extracts, although the cytokinin concentrations were significantly increased. Therefore, this study proved that the versatility of Methylobacterium as a plant-growth promoting bacteria could be better exploited.
Kang, Sang-Mo;Adhikari, Arjun;Lee, Ko-Eun;Khan, Muhammad Aaqil;Khan, Abdul Latif;Shahzad, Raheem;Dhungana, Sanjeev Kumar;Lee, In-Jung
Journal of Microbiology and Biotechnology
/
v.30
no.5
/
pp.717-725
/
2020
The use of plant growth-promoting rhizobacteria is economically viable and environmentally safe for mitigating various plant stresses. Abiotic stresses such as flood and drought are a serious threat to modern agriculture. In the present study, the indole-3-acetic acid-producing rhizobacterium R. sphaeroides KE149 was selected, and its effects on the growth of adzuki bean plants under flood stress (FS) and drought stress (DS) were investigated. IAA quantification of bacterial pure culture revealed that KE149 produced a significant amount of IAA. Moreover, KE149 inoculation notably decreased stress-responsive endogenous abscisic acid and jasmonic acid and increased salicylic acid in plants under DS and FS. KE149 inoculation also increased proline under DS and methionine under FS. In addition, KE149 inoculation significantly increased the levels of calcium (Ca), magnesium (Mg), and potassium (K) while lowering the sodium (Na) content in the plant shoot under stress. KE149-treated plants had markedly greater root length, shoot length, stem diameter, biomass, and higher chlorophyll content under both normal and stressed conditions. These results suggest that KE149 could be an efficient biofertilizer for mitigating water stress.
The effect of sodium sulphate on shoot induction and multiple shoot formation from nodal explants of Vitex negundo L. was tested on Murashige and Skoog's (MS) medium fortified with different auxins, cytokinins and sucrose. Highest percentage $(97.78\%)$ of explants for shoot induction and multiple shoot (20.68/explant) production were observed in the combination treatment of $N^6-Benzyl$ adenine (BA) $(17.80\;{\mu}M/L)$, ${\alpha}-Naphthalene$ acetic acid (NAA) $(2.15\;{\mu}M/L)$ and $5\%$ sucrose supplemented with 100 mg/L sodium sulphate. In vitro raised shoots were rooted on the half-strength MS medium fortified with different concentrations of NAA, Indole-3-acetic acid (IAA), and Indole-3-butyric acid (IBA) alone and in combinations. Among the treatments, $4.90\;{\mu}M/L$ of IBA was found most effective $(95.56\%)$ in inducing roots. The rooted plantlets were shifted to glasshouse for acclimatization and later transferred to the field with cent percent survival. Furthermore, in vitro flowering was observed in the shoots cultured on MS medium supplemented with BA $(8.90\;{mu}M/L)$ and NAA $(1.61\;{\mu}M/L)$.
The effects of IAA $(10^{-3}M)\;an;GA_3\;(2{\times}10^{-8}\;M)$ on the biosynthesis of phospholipids and their fatty acid compositions in chloroplast isolated from Chlorella ellipsidea were analyzed. Growth ratio and the levels of total lipids and phospholipids in whole cell system and chloroplast treated with phytohormones were higher than those of control, and when treated with GAl. phosphatidylcholine content was predominantly higher than that of control. It was showed that in whole cell system the fatty acid utilized for biosynthesis of phopholipids was 27.43% for palmitic acid in control, while that was 20.25% for linolenic acid in treatment with 1M. In treatment with $GA_3$, 23.17% for linolenic acid was used in phospholipid formation. The major fatty acid in the chloroplst system was analyzed to be 35.67% for palmitic acid in control and in treatment with 1M, 24.91% for linolenic acid was used in phospholipid formation, while in treatment with $GA_3$, major fatty acid of phospholipids was 22.80% for linoleic acid.c acid.
Philip Robinson, J.;Jyoti, Prasad Kakati;Sebastinraj, J.;Suriya, K
Journal of Plant Biotechnology
/
v.44
no.3
/
pp.343-348
/
2017
Cymbidium aloifolium (L). Sw. is an exquisite epiphytic orchid of the Kolli Hills (Eastern Ghats) of Tamil Nadu in Southern India. It is fast disappearing from its natural habitats due to deforestation and low germination rate in natural habitat. In the present study, an attempt was made to germinate the seeds from un-dehisced capsule of Cymbidium aloifolium (L). Sw under in vitro condition. The seed germination and protocorm development were recorded in three different well known media namely Knudson C (KC), Half strength Murashige & Skoog (1/2 MS) and Vacin & Went (VW) media. The highest seed germination of 90% was observed KC basal media after $30^{th}$ days whereas germination percentages were 40% and 30% on 1/2 MS and VW media respectively. The well-developed protocorm were transferred to KC media supplemented with 6-Benzyl Amino Purine (BAP) and Naphthalene acetic acid (NAA) where BAP (1.0 mg/l) and NAA (1.0 mg/l) together were found to be optimum for the highest shoot formation. About 90% of the shoots found to be well rooted after transfer to the KC medium differently supplemented with 1.5 mg/l Indole-3-acetic acid (IAA) and 1.0 mg/l Indole-3-butyric acid (IBA). Though rooting also took place in the two basic media but the duration was longer when compared with the hormone-supplemented media. The rooted plantlets were hardened and kept under greenhouse conditions which can be relocated in natural habitats.
Branched broomrape (Orobanche ramosa) is a holo-parasitic flowering plant that attaches to the root of its host, green plant, by means of a specialized structure known as haustorium. Following successful contact and penetration on susceptible plant root, complex tissue of Orobanche cells is formed which is known as the tubercle. Newly formed tubercles contain high activity ofindole-3-acetic acid (IAA). Triiodobenzoic acid (TIBA), as an inhibitor of IAA polar transport, was utilized to investigate the supply and requirement of auxin to the developing O. ramosa on tomato plant. There was no significant reduction in the incidence of O. ramosa per pot of different TIBA treatments. However, infection severity in terms of the number of O. ramosa shoots that emerged per plant and number of attachments per plant root system were significantly reduced by 60 % and 45 % on TIBA treated plants, respectively. Histo-logical studies revealed conspicuous delay in the initiation of xylem vessel differentiation inside tubercles of the TIBA treated tomato plants. Also, differentiated vessels showed thinner secondary wall deposition, and improper alignment within bundles inside those tubercles. They were wider and shorter in diameter in comparison to those of untreated plants. These findings were attri-buted to the short supply of IAA required for normal development, and to the xylem vessel differentiation of O. ramosa tubercles on infected tomato. Hence, this parasitic flowering plant seems to depend upon its host in its requirements for IAA, in a source to sink relation-ship.
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