• Title/Summary/Keyword: IBA(indole butyric 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.

Lateral Bud Suppression and Runner Plants Growth of 'Maehyang' Strawberry as Affected by Application Method and Concentration of IBA (IBA 처리방법과 농도에 따른 딸기 '매향'의 측아 발생 억제와 자묘 생육)

  • Hwang, Hee Sung;Jeong, Hyeon Woo;Lee, Hye Ri;Hwang, Seung Jae
    • Journal of Bio-Environment Control
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    • v.29 no.1
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    • pp.80-88
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    • 2020
  • This study was conducted to examine the effect by application method and concentration of the indole-3-butyric acid (IBA), which is auxin-based plant growth regulator, on the growth and runner plants production of strawberry in the greenhouse. The seedlings of strawberry were transplanted in the pot (150 ×135 × 90 mm) filled with coir medium on April 12, 2019. The IBA was applied with a foliar spray or drench as 50, 100, 150, and 200 mg·L-1 (50 mL per plant), respectively. The treatment was started on April 29, 2019. The foliar spray and drench treatment of IBA were repeated at 2-week intervals for 9 weeks from the start date of treatment. At 9 weeks after treatment, the petiole length of mother plants was the shortest in the control. The number of runner plants showed a tendency to decreased in the foliar spray. The number of lateral buds showed a tendency to decreased in the IBA treatment, and the least in the foliar with 100 mg·L-1. There was not significantly difference in the fresh and dry weights of the first and second runner plants. However, in the third runner plants, the fresh and dry weights were the greatest in the drench with 100 mg·L-1. Therefore, when considering the growth of third runner plants and lateral bud suppression, the drench with the 100 mg·L-1 could be better application method and concentration of IBA treatment for growth of the third runner plants and runner plants production of strawberry, and the results can be used as a basic research of plant growth regulator application to save the labor force and enhance the seedling quality in strawberry seedling stage.

In Vitro Propagation Through Nodal Explants in Helicteres isora L., a Medicinally Important Plant

  • Shriram, Varsha;Kumar, Vinay;Shitole, M.G.
    • Journal of Plant Biotechnology
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    • v.34 no.3
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    • pp.189-195
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    • 2007
  • Helicteres isora is medicinally important plant effective against asthma, diabetes, hypolipidemia, HIV, besides a good source of diosgenin. Seed dormancy and low rate of natural fruit production make this plant a perfect candidate for developing an in vitro method useful for its clonal propagation and further biotechnological developments. This is the first report on in vitro production of this plant. Nodal explants obtained from aseptically germinated seedlings were cultured on MS medium (Murashige and Skoog 1962) fortified with indole-3-acetic acid (IAA) ($0.57-22.83\;{\mu}M$), indole-3-butyric acid (IBA) ($0.41-16.58\;{\mu}M$), 6-benzylaminopurine (BA) ($0.44-17.75\;{\mu}M$) and kinetin (Kin) ($0.46-13.94\;{\mu}M$) either singly or in combinations of IAA + BA, IAA + Kin and BA + Kin. Combinations of cytokinins (BA and Kin) were most suitable for multiple shoot induction and $13.94\;{\mu}M\;Kin\;+\;13.31\;{\mu}M\;BA$ was optimum (79% frequency) associated with high number of microshoots (7.1 shoots per explant) after 20 days of culture. Maximum shoot elongation and proliferation (10 shoots per explant with 4.8 cm average height) was achieved on MS media containing $2.32\;{\mu}M\;Kin\;+\;2.22\;{\mu}M\;BA\;+\;2.85\;{\mu}M\;IAA$. High rooting frequency (70%) was achieved on MS medium (1/2 basal strength) fortified with $4.14\;{\mu}M$ IBA, while activated charcoal showed inhibitory effects on rooting. Hardening was done with 76% survival rate and these plants were growing without any visual defects and morphologically mimicking the naturally growing plants.

Growth, secondary metabolite production and antioxidant enzyme response of Morinda citrifolia adventitious root as affected by auxin and cytokinin

  • Baque, Md. Abdullahil;Hahn, Eun-Joo;Paek, Kee-Yoeup
    • Plant Biotechnology Reports
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    • v.4 no.2
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    • pp.109-116
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    • 2010
  • Morinda citrifolia adventitious roots were cultured in shake flasks using Murashige and Skoog medium with different types and concentrations of auxin and cytokinin. Root (fresh weight and dry weight) accumulation was enhanced at 5 $mg\;l^{-1}$ indole butyric acid (IBA) and at 7 and 9 $mg\;l^{-1}$ naphthalene acetic acid (NAA). On the other hand, 9 $mg\;l^{-1}$ NAA decreased the anthraquinone, phenolic and flavonoid contents more severely than 9 $mg\;l^{-1}$ IBA. When adventitious roots were treated with kinetin (0.1, 0.3 and 0.5 $mg\;l^{-1}$) and thidiazuron (TDZ; 0.1, 0.3 and 0.5 $mg\;l^{-1}$) in combination with 5 $mg\;l^{-1}$ IBA, fresh weight and dry weight decreased but secondary metabolite content increased. The secondary metabolite content (including 1,1-diphenyl-2-picrylhydrazyl activity) increased more in TDZ-treated than in kinetin-treated roots. Antioxidative enzymes such as catalase (CAT) and guaiacol peroxidase (G-POD), which play important roles in plant defense, also increased. A strong decrease in ascorbate peroxidase activity resulted in a high accumulation of hydrogen peroxide. This indicates that adventitious roots can grow under stress conditions with induced CAT and G-POD activities and higher accumulations of secondary metabolites. These results suggest that 5 $mg\;l^{-1}$ IBA supplementation is useful for growth and secondary metabolite production in adventitious roots of M. citrifolia.

High-frequency regeneration of plants in vitro from seedling-derived apical bud explants of Tilia mandshurica Rupr. & Maxim

  • Kim, Tae-Dong;Kim, Nam-Ho;Park, Eung-Jun;Lee, Na-Nyum
    • Journal of Plant Biotechnology
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    • v.48 no.1
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    • pp.54-61
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    • 2021
  • This work describe an efficient method for the shoot induction and plant regeneration of seedling-derived apical bud explants of Tilia mandshurica Rupr. & Maxim. The highest rate of shoot induction (82.2%) was obtained when apical bud explants from juvenile seedlings (5 months old) were cultured on Murashige and Skoog (MS) medium containing 1.0 mg/L 6-benzylaminopurine (BAP). However, apical bud explants obtained from mature trees (12 years old) did not produce any shoots, even with BAP supplementation. Among the three cytokinins tested for shoot multiplication (BAP, zeatin, and kinetin), BAP was the most effective; the highest number of shoots per explant (2.1) was observed on MS medium supplemented with 1.0 mg/L BAP. In contrast, the longest average shoot length (3.0 cm) was observed after growth on MS medium with 2.0 mg/L zeatin. No multiplication occurred when apical bud explants were cultured with kinetin-supplemented media. During rooting of in vitro-elongated shoots, the highest rooting rate (100%) was observed in half-strength MS medium supplemented with 0.5 ~ 1.0 mg/L indole-3-butyric acid (IBA) or 3.0 mg/L 1-naphthaleneacetic acid (NAA). During the acclimatization process, plantlets that were rooted on the IBA (0.5 mg/L)-supplemented medium had the highest survival rate (100%) and maximum root length (18.5 cm). These findings suggest that a low concentration (0.5 mg/L) of IBA is appropriate for the rooting and acclimatization of T. mandshurica. Plants were successfully transferred to the greenhouse with a 100% survival rate. This protocol will be useful for the large-scale propagation of Tilia species.

Determination of Optimal Collecting Date and Exogenous Auxin Dipping Treatments in Cutting Transplants of 'Seolhyang' Strawberry (Fragaria × ananassa Duch.) ('설향' 딸기 삽목묘의 최적 삽수 채취시기와 오옥신 처리 구명)

  • Kim, Eun Ji;Uhm, Mi Jeong;Jung, Hyun Soo;Kim, Jong Yeob;Lee, Jun Gu
    • Journal of Bio-Environment Control
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    • v.29 no.3
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    • pp.252-258
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    • 2020
  • The aim of this study was to investigate the effects of timing of collecting date and concentration of IBA and NAA, in order to enhance initial activity and seedling quality of domestic strawberry. Strawberry cuttings were separately taken twice, in June 7 and in July 5, and IBA and NAA were treated with the concentrations of 0.025, 0.05 and 0.1% at cutting date, respectively. The seedlings were evaluated for the percentage of survival during 18 days at 6 times after tunnel cultivation. The NAA treatment was inappropriate for strawberry cutting due to the high rate of seedling mortality, regardless of the collecting date. The vitality of the seedlings was highest at IBA 0.1% in June collecting and at IBA 0.05% in July collecting. The seedlings from June collecting had a higher quantum yield at IBA 0.1% and the seedlings from July collecting at IBA 0.05%. Therefore, IBA could be more effectively applied than NAA to promote the vitality and quality with the appropriate concentration of 0.1% at June collecting and 0.05% at July collecting, respectively.

Cutting Propagation of Dendropanax morbifera $L_{EV}$. (황칠나무 삽목번식에 관한 연구)

  • Choi, Seong-Kyu
    • Korean Journal of Medicinal Crop Science
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    • v.6 no.4
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    • pp.251-257
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    • 1998
  • This experiment was carried out to establish cutting propagation method of dendropanax morbifera $L_{EV}\$. at Wando in Chonnam, native area. The hardwood cutting and the greenwood cutting were able to be used as propagation method, but callus formation and rooting ratio in the greenwood cutting were higher than in hardwood cutting. The optimum cutting time was February to middle of March in hardwood cutting and July to August in greenwood cutting. The earthen-ball cutting method was better than normal cutting method in callus formation and rooting ratio. The rooting in different bed soils was the best at sand-loam soil. The application of IBA 100ppm promoted rooting.

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Factors Affecting on Regeneration in 'WHANGKEUMBAE' Pear (Pyrus pyrifolia)

  • Lee, Gung-Pyo
    • Journal of agriculture & life science
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    • v.43 no.6
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    • pp.59-65
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    • 2009
  • Combinations of plant growth regulators, darkness treatments, and the order of expanding leaves for explants were evaluated for optimizing in vitro shoot regeneration rate of 'Whangkeumbae' pear. In a MS medium, supplemented with $8.88{\mu}M$ 6-benzylaminopurine (BA) and $0.49{\mu}M$ indole-3-butyric acid (IBA), green foci were observed on the surface of the callus 8 days after culture initiation. Some adventitious buds were later induced from those green foci, resulting in the appearance of normal shoots. In a medium containing $22.20{\mu}M\;BA$, the surface of the callus became compact and greenish, and many adventitious buds were formed over the entire area of the callus surface. When comparing BA concentration via histological observation, the section which had been treated with $22.20{\mu}M\;BA$ exhibited closer cell aggregation than those with $8.88{\mu}M\;BA$. The darkness treatment enhanced the formation of adventitious shoots for up to 3 weeks. The youngest two expanding leaves, proximal to the shoot apex, were proved to be the most regenerative, and yielded the highest shoot number per regenerating leaf. A fourth strength MS medium, which was supplemented with $0.54{\mu}M\;NAA$, yielded good quality plantlets, with regard to root number and root length.

In vitro Root Induction from Shoot Explants of Pear (Pyrus spp.)

  • Jae-Young Song;Jinjoo Bae;Woohyung Lee;Jung-Ro Lee;Mun-Sup Yoon
    • Korean Journal of Plant Resources
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    • v.35 no.6
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    • pp.770-777
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    • 2022
  • The main objective of this study was to identify the most appropriate condition for root formation of in vitro micropropagated pear (Pyrus spp.) plants. In vitro propagation was induced on Murashige and Skoog (MS) medium with 2.0 mg/L of N6-benzyladenine (BA) and 0.2 mg/L of Indole-3-butyric acid (IBA) medium. The short pre-treatment of explants with a high concentration (1 mg/L) of NAA and IBA (R0 medium) in dark for three days, followed by transfer to five different media (R1 to R5) resulted in good rooting responses in the pear 'Oharabani (P. pyrifolia × P. communis)' genotype. For the rooting experiments, the highest rooting percentage (83.3 ± 8.3%), average root length (3.6 ± 1.9 mm), total root number (31 ± 4.0), and average root number per plant (2.6 ± 2.1) were obtained on half strength (1/2) of MS medium supplemented with 30 g/L sucrose without hormones and activated charcoal (AC) (R1 medium). The highest rooting percentage was obtained at 83.3% from explants on R1 and R3 media. The rooting procedure described in this study resulted in good root formation and significantly shorting the root induction time to within 14 days of culture. Further studies are underway to test the suitability of the protocol developed in this study for other pear genotypes.