• Title/Summary/Keyword: IAA (indole acetic acid)

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Efficient and Reliable in vitro Regeneration System for Rubus Species as the Basis of Genetic Engineering

  • Kalai Katalin;Meszaros Annamaria;Denes Ferenc;Zatyko Jozsef;Balazs Ervin
    • Journal of Plant Biotechnology
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    • v.7 no.4
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    • pp.241-246
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    • 2005
  • Factors affecting regeneration of different Rubus varieties (blackberry, raspberry and their hybrid) were examined and a reliable regeneration system was established. Media for stock plant maintenance were tested; different explants and media were investigated to find the best circumstances for the regeneration. The effect of the commonly used antibiotics was studied to determine the most suitable one for selection of the transformants. We found that both MS and LS media supplemented by $20\;gL^{-1}$ sucrose are suitable for the stock plant maintenance. The optimal hormone content for the stock plants is $0.125\;mgL^{-1}$ 6-benzylaminopurine (BAP) with $0.01\;mgL^{-1}$ indole-3- butyric acid (IBA). The highest regeneration rate was observed on medium containing MS salts with B5 vitamins complemented with glucose, sucrose, maltose, $10\;gL^{-1}$ each, supplemented with benzylaminopurine riboside (BAR) ($2\;mgL^{-1}$) and indole-3-acetic acid (IAA) ($0.1\;mgL^{-1}$). The regenerated shoots appeared directly from the cut edges, without callus phase. Hygromycin and geneticin proved to be good selection agents for the Rubus explants, but due to their severe effect on the tissues we propose to use marker-free constructions for the transformation.

An Improved Method of Organogenesis from Cotyledon Callus of Acacia sinuata (Lour.) Merr. using Thidiazuron

  • Shahzad Anwar;Ahmad Naseem;Anis Mohammad
    • Journal of Plant Biotechnology
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    • v.8 no.1
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    • pp.15-19
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    • 2006
  • An efficient protocol for in vitro multiple shoot bud induction and plant regeneration from mature green cotyledon derived callus tissues of Acacia sinuata has been developed. Callus formation occurs at all the concentrations of thidiazuron (TDZ) in Murashige and Skoog's (MS) medium, but 0.6 ${\mu}M$ proved to be the best with maximum callus formation frequency. Supplementation of TDZ in combination with indole-acetic acid (IAA) in MS media accelerates shoot bud organogenesis in differentiating callus tissues with 60-70% conversion of shoot buds into shoot Most efficient shoot organogenesis was recorded when TDZ induced calli were subcultured at different concentrations of 6-benzyla-denine (BA). Optimum shoot bud induction and plant regeneration from callus was achieved when 0.6 ${\mu}M$ (TDZ) induced calli were subcultured at 3.0 ${\mu}M$ (BA) where $16.6{\pm}0.74$ shoots/unit callus on obtained. Rooting in in vitro differentiated shoots was achieved when transferred to medium containing different concentration of indole-3-butyric acid (IBA) in full & half strength MS medium. The well rooted plantlets were hardened and transferred to net house with 90% survival rate.

Effects of Cold Pretreatment and Medium Composition on Anther Culture Initiation in Strawberry

  • Na, Hae-Young;Kim, Dae-Young;Chun, Chang-Hoo
    • Horticultural Science & Technology
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    • v.29 no.5
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    • pp.488-493
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    • 2011
  • Callus culture initiation of strawberry (Fragaria${\times}$ananassa Duch.) was investigated at different Murashige and Skoog (MS) medium strengths, types and concentrations of plant growth regulators, and incorporating a cold pretreatment period to determine the optimal nutritional and environmental conditions. No high quality callus was induced on MS media without auxin regardless of medium strength. When 6-benzylaminopurine (BA) was combined with indole acetic acid (IAA), naphthalene acetic acid (NAA), and 2,4-dichlorophenoxyacetic acid (2,4-D), high quality callus were highly induced compared to medium supplemented with auxin alone. When $0.5mg{\cdot}L^{-1}$ BA was combined with IAA, NAA, and 2,4-D, high quality callus induction was more effective than the medium supplemented with the other BA concentrations. The best combination of auxin and cytokinin for high quality callus induction was $1.0mg{\cdot}L^{-1}$ NAA and $0.5mg{\cdot}L^{-1}$ BA. Although the differences in callus induction were not significant, high quality callus induction at half strength MS medium was more effective than at full strength medium. When $30g{\cdot}L^{-1}$ sucrose was added to the half strength MS medium, the rate of high quality callus induction increased. The optimum cold pretreatment temperature and period for high quality callus induction were $4^{\circ}C$ and 72 h, respectively. Regeneration rate of high quality callus increased in MS medium supplemented with thidiazuron.

Isolation of Photosynthetic Bacterium, Rhodopseudomonas palustris JK-1 and Researches on IAA and Carotenoid Production (광합성세균 Rhodopseudomonas palustis 분리 및 IAA와 Carotenoid 생성에 관한 연구)

  • Kim, Yu-Kyoung;Cho, Young-Yun;Kang, Ho-Jun;Kim, Jung-Sun;Yang, Sung-Nyun;Jwa, Chang-sook
    • Korean Journal of Organic Agriculture
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    • v.25 no.4
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    • pp.843-859
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    • 2017
  • The JK-1 isolate which was the best producer of indole-3-acetic acid and carotenoid among the 388 strains isolated from 28 wetlands in Jeju, was identified to be Rhodopseudomonas palustirs belongs to a typical group of non sulfur purple bacteria based on 16S sRNA sequencing. This study investigated the effect of different cultural conditions of pH, temperature, agitation, light and aeration on growth, IAA and carotenoid production of photosynthetic bacterium JK-1 for optimization of IAA and carotenoid production. It was found that growth, IAA, carotenoid, and bacteriochlorophyll production with light (3,000~3,500 Lux) and agitation (100 rpm) showed better results than those with dark/static or dark/agitation (100 rpm) in anaerobic conditions. The optimal pH, temperature and agitation speed for cell growth were 7, $30^{\circ}C$, 150 rpm, for IAA production were 9, $30^{\circ}C$, 150rpm and for carotenoid production were 6, $25^{\circ}C$, 50 rpm, cultured for 72 h under anaerobic light, respectively. The growth and IAA production were high in aerobic culture compared with anaerocic culture, whereas carotenoid and bacteriochlorophyll content were decreased extremely in aerobic condition (0.5~1 vvm). Subsequently, the optimal culture conditions for JK-1 were selected with pH 7, $30^{\circ}C$ and 100 rpm under anaerobic light and the effect on plant growth was tested by pot assay. Inoculation of JK-1 with 3% (v/v) level caused increase in shoot and root dry weigh that varied from 20%~58% to 40%~28% in young radish in camparison to uninoculated treatment at 50 days of growth. The study suggests that the JK-1 isolate may serve as efficient biofertilizer inoculants to promote plant growth.

Clonal Propagation in Commiphora Wightii (Arnott.) Bhandari

  • Mishra, Dhruv Kumar;Kumar, Devendra
    • Journal of Forest and Environmental Science
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    • v.30 no.2
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    • pp.218-225
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    • 2014
  • Studies were carried out to standardize and develop a suitable macro-propagation technology for large scale production of superior clonal stock through stem cuttings in Commiphora wightii Arnott (Bhandari), a data deficient medicinal plant of arid region. For the purpose, three experiments were conducted. The first experiment was tried to elucidate the impact of various cutting diameters (0.50-0.75 cm, 0.75-1.00 cm, 1.00-1.50 cm, and >1.50 cm) in combination with varying growing conditions (sunlight, shade house and mist chamber) on shoot sprouting and rooting without using exogenous plant growth regulators. Cutting diameter (size 0.75-1.00 cm) in mist chamber has shown maximum sprouting (90.00%) and rooting (73.33%), primary root (6.67) and secondary root (16.67) followed by 1.00-1.51 cm in mist chamber. Minimum sprouting (40.00%), rooting (33.33%), number of shoot (1.33), primary root (1.00) and number of secondary root (1.00) was recorded in cutting diameter (size >1.50 cm) in sunlight. Second experiment was performed to find out optimum growth regulator concentration of rooting hormone (100, 200, 500 and 1000 ppm) of Indole-3-acetic acid (IAA) and Indole-3-butyric Acid (IBA) on adventitious root formation on cuttings diameter (size 0.25-0.50 cm) in comparison to control. Maximum rooting percentage (93.33%) was recorded in 200 ppm followed by 500 ppm (86.66%) of IBA as compared to control, which showed only 60 per cent sprouting. Third experiment was performed with newly formed juvenile micro-cuttings treated with varying concentrations of IAA and IBA. The juvenile cuttings (size 6-10 cm, basal dia <0.25 cm) were selected as micro-cuttings. The cuttings treated with IBA (500 ppm) showed 64.30% rooting as compared to other treatments. Results of above experiments indicate that cuttings (size 0.75-1.00 cm dia) may be developed in mist chamber for better performance. While using heavier cuttings, no growth promoting hormones is required however; growth regulator 200 ppm concentration of IBA rooting hormone was observed optimum for promoting macro-propagation in stem cuttings of lower diameter class (0.25-0.50 cm).

Plant growth promotion effect of Klebsiella michiganensis Jopap-1 isolated from button mushroom bed (양송이배지로부터 분리한 Klebsiella michiganensis Jopap-1의 식물생장촉진효과)

  • Kim, Ye-Seul;Yoon, Min-Ho
    • Journal of Mushroom
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    • v.16 no.3
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    • pp.218-224
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    • 2018
  • An auxin-producing bacterium, Klebsiella michiganensis Jopap-1, was isolated from a button mushroom bed in Buyeo-Gun, Chungcheongnam-Do. The strain Jopap-1 was classified as a novel strain of K. michiganensis based on a chemotaxonomic and phylogenetic analysis. The isolated K. michiganensis Jopap-1 was confirmed to produce indole-3-acetic acid (IAA), which is one of auxin hormones by TLC and HPLC analyses. The maximum concentration of IAA ($96.05mg\;L^{-1}$) was detected in the culture broth incubated in R2A medium containing 0.1% L-tryptophan for 48 h at $35^{\circ}C$ by HPLC quantity analysis. A negative relationship between IAA production and pH variation was estimated to show that the increase of IAA caused acidic pH in the culture. The effect of the supplement on L-tryptophan (precursor of IAA) production was observed to be highest at 0.1% concentration, but was significantly lowered above a concentration of 0.2%. To investigate the growth-promoting effects on the crops, the culture broth of E. michiganensis Jopap-1 was infected to water cultures and seed pots of mung bean and lettuce. Consequently, the adventitious root induction and root growth of mung bean and lettuce were observed to be 2.1 and 1.8 times higher than those of the control.

Interaction of Brassinolide with Other Known Plant Growth Regulators (Brassinolide와 기존 식물생장조절제(植物生長調節劑)와의 상호작용(相互作用))

  • Choi, C.D.;Takematsu, T.;Takeuchi, Y.;Kim, K.U.
    • Korean Journal of Weed Science
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    • v.7 no.1
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    • pp.78-83
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    • 1987
  • This study was attempted to evaluate the combining effect of HBR (homobrassinolide) with the known growth regulators such as GA (gibberellic acid), BA(6-benzyl aminopurine), IAA (indole-3-acetic acid), B-9 (N-dimethylamino succinamic acid) and CCC (2-chloroethyl-trimethylammonium chloride) on the growth of radish hypocotyl. A single application of HBR increased hypocotyl growth as its rates increased from 0.1 to 1.0 ppm, showing a maximum increase at 1.0 ppm. GA and BA had no direct effects on hyopcotyl growth, but IAA showed some effect as its concentration increased. However, the mixed application of HBR with GA, BA and IAA increased the length of radish hypocotyl as the concentration of HBR became higher. The mixture of HBR with GA and BA showed antagonistic reaction on radish hypocotyl growth, but synergistic effect was shown in the higher rate mixture of HBR with IAA in the range of HBR at 0.03 to 0.30 ppm with IAA at 3.0 to 10.0 ppm, but antagonistic or additive response at the mixture of low rates. An increased growth of hypocotyl by HBR was ified by CCC, showing the strong antagonistic reaction, but B-9 was not able to ify HBR's effect on hypocotyl growth.

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Physiological Responses of Rice Seedlings to Butachlor (Butachlor에 대한 벼 유묘의 생리적 반응)

  • Tsai, Wen-Fu
    • Korean Journal of Weed Science
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    • v.15 no.4
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    • pp.247-253
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    • 1995
  • The herbicide butachlor [N-(butoxymethyl)-2-chloro-N-(2,6-di-methylphenyl) acetamide] is widely used by farmers as a tool for weed management of transplanted rice(Oryza sativa L.) in Taiwan. The herbicide did not stop germination of rice and weed seeds, but strongly inhibited the subsequent growth of young shoots and roots. The inhibition was also strong on established seedlings. However, they could recover to normal growth after the herbicide effect disappeared. Butachlor greatly decreased the endogenous indole-3-acetic acid (IAA) but increased the endogenous abscisic acid (ABA) contents of rice seedlings. Addition of lAA into growth medium (Hoagland's solution) partly relieved growth inhibition. Pretreatment of both gibberellic acid ($GA_3$) and IAA 24 hours before butachlor treatment almost completely alleviated the butachlor-interfere with GA and/or IAA metabolism or their action resulting in the growth inhibition of rice. Butachlor was readily absorbed by rice roots. During 24 hours of uptake experiment, 32% of the applied herbicide was absorbed. Pretreatment of the herbicide for 2 days did ncx affect the absorption. Of the absorbed herbicide, 80% remained in roots, only 20% transported into shoots, and more than 50% was metabolized to water soluble substances. Thin-layer chromatographic (TLC) analysis indicated that the Rf value of the most abundant metabolite was butachlor-glutathione conjugate. Rice, barnyardgrass (Echinochloa crus-galli (L.) Beauv.), and monochoria (Monochoria vaginalis Presl) seedlings contained relatively high level of non-protein thiols, while the glutathione S-transferase (GST) activity was found highest in rice, barnyardgrass the next, monochoria the lowest. The difference in GST activity among these species might be related to their sensitivity to butachlor.

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Characterization of a Heavy Metal-Resistant and Plant Growth-Promoting Rhizobacterium, Methylobacterium sp. SY-NiR1 (중금속 내성 및 식물 생장 향상 근권세균 Methylobacterium sp. SY-NiR1의 분리 및 특성)

  • Koo, So-Yeon;Cho, Kyung-Suk
    • Microbiology and Biotechnology Letters
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    • v.35 no.1
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    • pp.58-65
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    • 2007
  • The role of soil microorganisms, specifically rhizobacteria, in the development of rhizoremediation techniques is important to speed up the process and to increase the rate of mobilization or absorption of heavy metals to the plant. In this study, Methylobacterium sp. SY-NiR1 was isolated from the rhizosphere soils of plants in oil and heavy metal-contaminated soil. Based on its pink pigmented colony, rod-shape cells, and belonging in $\alpha-Proteobacteria$, Methylobacterium sp. SY-NiR1 is considered a pink-pigmented facultative methylotroph. SY-NiR1 had the ability to produce indole acetic acid which is one of phytohormones. This bacterium showed resistance against multiple heavy metals such as Cd, Cr, Cu, Pb, Ni, Zn, and the order of its resistance based on $EC_{50}$ was Zn > Ni > Cu > Pb > Cd > Cr. Therefore, Methylobacterium sp. SY-NiR1 can stimulate seed germination and plant growth in soil contaminated with heavy metals.

Nodulation Experiment by Cross-Inoculation of Nitrogen-Fixing Bacteria Isolated from Root Nodules of Several Leguminous Plants

  • Ahyeon Cho;Alpana Joshi;Hor-Gil Hur;Ji-Hoon Lee
    • Journal of Microbiology and Biotechnology
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    • v.34 no.3
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    • pp.570-579
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    • 2024
  • Root-nodule nitrogen-fixing bacteria are known for being specific to particular legumes. This study isolated the endophytic root-nodule bacteria from the nodules of legumes and examined them to determine whether they could be used to promote the formation of nodules in other legumes. Forty-six isolates were collected from five leguminous plants and screened for housekeeping (16S rRNA), nitrogen fixation (nifH), and nodulation (nodC) genes. Based on the 16S rRNA gene sequencing and phylogenetic analysis, the bacterial isolates WC15, WC16, WC24, and GM5 were identified as Rhizobium, Sphingomonas, Methylobacterium, and Bradyrhizobium, respectively. The four isolates were found to have the nifH gene, and the study confirmed that one isolate (GM5) had both the nifH and nodC genes. The Salkowski method was used to measure the isolated bacteria for their capacity to produce phytohormone indole acetic acid (IAA). Additional experiments were performed to examine the effect of the isolated bacteria on root morphology and nodulation. Among the four tested isolates, both WC24 and GM5 induced nodulation in Glycine max. The gene expression studies revealed that GM5 had a higher expression of the nifH gene. The existence and expression of the nitrogen-fixing genes implied that the tested strain had the ability to fix the atmospheric nitrogen. These findings demonstrated that a nitrogen-fixing bacterium, Methylobacterium (WC24), isolated from a Trifolium repens, induced the formation of root nodules in non-host leguminous plants (Glycine max). This suggested the potential application of these rhizobia as biofertilizer. Further studies are required to verify the N2-fixing efficiency of the isolates.