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1

Arif, Mahmoud B., and Houchang Khatamian. "IN VITRO EMBRYOGENESIS DERIVED FROM LEAF CALLUS OF `TIFFANY' ROSE." HortScience 25, no. 9 (September 1990): 1085G—1086. http://dx.doi.org/10.21273/hortsci.25.9.1085.

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Friable callus from leaf disks of Rosa hybrida `Tiffany' was initiated within two weeks under dark conditions and 25°C on Murashige and Skoog (MS) medium supplemented with 4 mg.liter-1 2,4-D. Callus was then transferred into MS medium containing 3 mg.liter-1 2,4-D. Within four weeks, rhizogenesis occurred on the callus surface. The rhizogenic calllus was subculture on MS medium plus 3 mg.liter-1 2,4-D every 4-6 weeks. Within six months from initial culture, somatic embryos were developed on the aging callus in darkness. Transfer of the aging callus with somatic embryos into 1/2 MS medium containing 1 mg.liter-1 kinetin and maintaining it under 46 μE m-2s-1 light for 16 hrs. resulted in greening of the somatic embryos.
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2

Arif, Mahmoud B., and Houchang Khatamian. "IN VITRO EMBRYOGENESIS DERIVED FROM LEAF CALLUS OF `TIFFANY' ROSE." HortScience 25, no. 9 (September 1990): 1085g—1086. http://dx.doi.org/10.21273/hortsci.25.9.1085g.

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Friable callus from leaf disks of Rosa hybrida `Tiffany' was initiated within two weeks under dark conditions and 25°C on Murashige and Skoog (MS) medium supplemented with 4 mg.liter-1 2,4-D. Callus was then transferred into MS medium containing 3 mg.liter-1 2,4-D. Within four weeks, rhizogenesis occurred on the callus surface.The rhizogenic calllus was subculture on MS medium plus 3 mg.liter-1 2,4-D every 4-6 weeks. Within six months from initial culture, somatic embryos were developed on the aging callus in darkness.Transfer of the aging callus with somatic embryos into 1/2 MS medium containing 1 mg.liter-1 kinetin and maintaining it under 46 μE m-2s-1 light for 16 hrs. resulted in greening of the somatic embryos.
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3

Dainty, A. L., K. H. Goulding, P. K. Robinson, I. Simpkins, and M. D. Trevan. "Callus conclusion." Trends in Biotechnology 3, no. 9 (September 1985): 219. http://dx.doi.org/10.1016/0167-7799(85)90010-1.

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4

Gannon, Francis H., and Lester D. R. Thompson. "Traumatic Fracture Callus." Ear, Nose & Throat Journal 86, no. 4 (April 2007): 200. http://dx.doi.org/10.1177/014556130708600407.

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5

Oni, O. A. "The bony callus." Injury 28, no. 9-10 (November 1997): 629–31. http://dx.doi.org/10.1016/s0020-1383(97)00126-5.

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6

Toker, G., M. Memişoğlu, M. C. Toker, and E. Yeşilada. "Callus formation and cucurbitacin B accumulation in Ecballium elaterium callus cultures." Fitoterapia 74, no. 7-8 (December 2003): 618–23. http://dx.doi.org/10.1016/s0367-326x(03)00165-5.

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7

Dovzhenko, Alexander, and Hans-Ulrich Koop. "Sugarbeet ( Beta vulgaris L.): shoot regeneration from callus and callus protoplasts." Planta 217, no. 3 (July 1, 2003): 374–81. http://dx.doi.org/10.1007/s00425-003-1006-7.

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8

Famelaer, I., E. Ennik, W. Eikelboom, J. M. Van Tuyl, and J. Creemers-Molenaar. "The initiation of callus and regeneration from callus culture ofTulipa gesneriana." Plant Cell, Tissue and Organ Culture 47, no. 1 (February 1996): 51–58. http://dx.doi.org/10.1007/bf02318965.

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9

Qin, Yan Jun, Wu Que Gong, Ting Liu, Jing Hui Yang, Yan Jun Liu, and Wei Zi Huang. "Difference of Morphological Structure and Characteristics of Physiology and Biochemistry between Two Types of Alfalfa Callus." Applied Mechanics and Materials 700 (December 2014): 306–9. http://dx.doi.org/10.4028/www.scientific.net/amm.700.306.

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In order to know the key factors of callus development on single cell clone, characteristics of physiology and biochemistry and morphological structure on two types of Alfalfa callus was studied. The results showed that the cell membrane permeability and malondialdehyde content was same on two types of callus; but, their soluble sugar content and peroxidase activity in cells was larger different, the soluble sugar content: soft callus > huge callus, and soft callus was 2.09 times of huge callus; peroxidase activity: huge callus > soft callus, and huge callus was 1.35 times of soft callus. Huge callus cells were larger, loosely arranged, cell vacuolization and cytoplasm was thinner than soft callus. However, soft callus cells were smaller, loosely arranged, cell vacuoles was smaller and cytoplasm was thicker than hug callus. Huge cells had the same membrane metabolic with soft cells, but soft cells had higher sugar accumulation than huge cells, and soft cells metabolism are vigorous, while huge cells are more aging.
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10

Hu, Yan Yan, Jing Hui Yang, Chao Zhang, Ting Liu, Bin Li, and Ying Wang. "Difference of Cell Morphology on Different Callus Types of Alfalfa." Applied Mechanics and Materials 707 (December 2014): 137–43. http://dx.doi.org/10.4028/www.scientific.net/amm.707.137.

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In order to induce embryo from callus and set up the integral somatic embryo induction system from monoclonal cell, the differences of cell morphology and structure from different types of alfalfa callus were observed and compared by quick section and microscopic examination. The results show that giant callus cells were larger, elongated and yellow-white; loose callus cells were smaller, spherical, soft and yellow-green; hard-type callus cells were round, hard and dark green. The cell volume of giant callus was 4.5 times more than loose callus cells and 9 times more than hard-type callus cells. The biggest change of vacuole number and form were giant callus cells, which had 48%cells of 5-8 big vacuoles. Loose callus cells had 89%cells of 2-4 small vacuoles and hard-type callus cells had 97% cells with one large central vacuole. Loose callus cells had more chloroplast, which were 4.65 times more than giant callus cells. The chloroplast of hard-type callus cells was gathered into groups, which had 3-5 chloroplasts in it. The most nucleuses of giant callus cells and loose callus cells were located in the central of cell and 96.8% nucleus of hard-type callus cells were located on the edge. In hard-type callus cells there were different number of rings, thread and textured ducts. In short, there were lower cell differentiation and clearer vacuolization in giant callus, and high degree of differentiation and tissue aging in hard-type callus. The loose callus was undifferentiated, was lower on vacuolization and apparent on characteristics of embryonic callus, so that it was more suitable for induction of somatic embryos.
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11

VALLES, M., and Ph BOXUS. "REGENERATION FROM ROSA CALLUS." Acta Horticulturae, no. 212 (September 1987): 691–96. http://dx.doi.org/10.17660/actahortic.1987.212.118.

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12

IKUTA, Akira. "Callus Constituents and Chemotaxonomy." Plant tissue culture letters 4, no. 1 (1987): 1–7. http://dx.doi.org/10.5511/plantbiotechnology1984.4.1.

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13

Benson, J. Christopher, and Alan S. Banks. "First Metatarsal Callus Distraction." Journal of the American Podiatric Medical Association 98, no. 1 (January 1, 2008): 51–60. http://dx.doi.org/10.7547/0980051.

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We describe the results in seven patients who underwent surgery to lengthen a short first metatarsal via callus distraction. The increased length achieved ranged from 13 to 48 mm, with an average of 20.2 mm. The technique was successful in restoring length and improving symptoms, although several complications were encountered. (J Am Podiatr Med Assoc 98(1): 51–60, 2008)
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14

Gonzalez Benito, E., and P. G. Alderson. "REGENERATION FROM ALSTROEMERIA CALLUS." Acta Horticulturae, no. 280 (July 1990): 135–38. http://dx.doi.org/10.17660/actahortic.1990.280.21.

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15

Ellem, Bradon, and Russell Lansbury. "Tribute to Ron Callus." Journal of Industrial Relations 49, no. 2 (April 2007): 147–49. http://dx.doi.org/10.1177/0022185607074906.

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16

Palma, L. de, A. Tulli, G. Maccauro, S. P. Sabetta, and M. del Torto. "Fracture Callus in Osteopetrosis." Clinical Orthopaedics and Related Research &NA;, no. 308 (November 1994): 85???89. http://dx.doi.org/10.1097/00003086-199411000-00014.

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17

Jaramillo, J., and W. L. Summers. "Tomato Anther Callus Production: Solidifing Agent and Concentration Influence Induction of Callus." Journal of the American Society for Horticultural Science 115, no. 6 (November 1990): 1047–50. http://dx.doi.org/10.21273/jashs.115.6.1047.

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Anthers from. three tomato cultivars, `L-680A', `Ailsa Craig', and `Licato', were plated on DBM1 medium solidified with one of four solidifying agents, Bacto-agar, Gelrite, Noble agar, or Phytagar, to evaluate their ability to promote initiation and growth of tomato anther callus. The optimum concentration of each solidifying agent was compared with a liquid control. Optimum levels of the various solidifying agents were (in g·liter-1) Phytagar, 5; Gelrite, 3; Noble agar, 6 and Bacto-agar, 8. Both the number and diameter of calluses were affected by type of solidifying agent and anther genotype. Significant interactions were also found between tomato cultivars and solidifying agent. Noble agar produced good results with `L-680A' and `Ailsa Craig', but not with `Licato'. Bacto-agar reduced the number and size of callus by 38% when compared with the liquid treatment and by 42% when compared with the best agar treatment (Noble agar).
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18

ABE, Toshinori, and Yuzo FUTSUHARA. "Diallel analysis of callus growth and plant regeneration in rice seed-callus." Japanese Journal of Genetics 66, no. 2 (1991): 129–40. http://dx.doi.org/10.1266/jjg.66.129.

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19

Prem, Deepak, Subhadra Singh, Padam Prakash Gupta, Jaivir Singh, and Sish Pal Singh Kadyan. "Callus induction and de novo regeneration from callus in Guar (Cyamopsis tetragonoloba)." Plant Cell, Tissue and Organ Culture 80, no. 2 (February 2005): 209–14. http://dx.doi.org/10.1007/s11240-004-0738-9.

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20

Khairan, Khairan, Betty Mauliya Bustam, Yunita, Riska Meilinda, and Raudhatul Muna. "Pengaruh Komposisi 2,4-D dan BAP Terhadap Pembentukan Kalus Eksplan Pucuk Nilam (Pogostemon Cablin Benth.) secara In Vitro dengan Pemotongan Horizontal dan Vertikal." Biotropic : The Journal of Tropical Biology 5, no. 1 (February 27, 2021): 9–20. http://dx.doi.org/10.29080/biotropic.2021.5.1.9-20.

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This study aims to determine the effect of 2,4-Diclorophenoxy Acetic Acid (2,4-D) and Benzyl Amino Purin (BAP) on the formation of callus of patchouli (Pogostemon cablin Benth.) shoot explants by horizontal and vertical cutting methods. The parameters that observed in this study were the percentage growth of callus, time appearance of callus, weight of callus and the morphology of callus. The results showed that horizontal cutting method was able to induce callus growth with the percentages growth of callus were 18,75%, with the time appearance of callus was at 16 days at P1; P10; P12; P13 dan P14. The highest weight of callus obtained was 0.19 grams at P8. The results also showed that the callus yielded had a yellow and cream color, with a compact and crumb textures. Meanwhile, the vertical cutting method was able to induce callus formation with the percentage growth of callus were 12,5%. The fastest time of callus appearance was obtained in P6 and P8, which was 12 day after planting with the highest weight of callus obtained was 0.05 grams at P12. The results also showed that vertical cutting method had brown and dark-brown of callus with a compact and crumb textures.
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21

Chée, Raymond P., Daniel I. Leskovar, and Daniel J. Cantliffe. "Optimizing Embryogenic Callus and Embryo Growth of a Synthetic Seed System for Sweetpotato by Varying Media Nutrient Concentrations." Journal of the American Society for Horticultural Science 117, no. 4 (July 1992): 663–67. http://dx.doi.org/10.21273/jashs.117.4.663.

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Embryogenic callus growth of sweetpotato [Ipomoea batatas (L.) Lam.] was selectively enhanced by subculture on basal callus proliferation medium modified to contain 15 mm NH4NO3. Embryogenic callus production was doubled on basal callus proliferation medium modified to contain 60 mm K+, while nonembryogenic callus production was reduced 40%. Additions of up to 40 mm NaCl to basal callus proliferation medium did not affect callus proliferation. The development of embryos from calli subculture to embryo production basal medium was unaffected by the KCl or NaCl treatments of the callus proliferation phase. However, embryo production was increased by subculturing callus from callus proliferation medium containing 20 mm NH4+ to embryo production medium containing 10 mm NH4+ Our results demonstrate that changes in mineral nutrition, in addition to growth regulator differences between callus proliferation and embryo production media, are important factors in sweetpotato somatic embryogenesis.
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22

Fransz, P. F., and J. H. N. Schel. "Cytodifferentiation during the development of friable embryogenic callus of maize (Zea mays)." Canadian Journal of Botany 69, no. 1 (January 1, 1991): 26–33. http://dx.doi.org/10.1139/b91-005.

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Immature embryos of Zea mays L. were cultured on N6 medium to obtain embryogenie callus. Cultured tissue fragments with various developmental stages of friable callus were sampled and prepared for light microscopy and transmission electron microscopy. Pieces of compact callus were also prepared for light microscopy to compare the structural organization of both callus types. Friable callus develops from a thin layer of abaxial scutellum cells, including the epidermis. During further development the callus cells dissociate, owing to the breakdown of the middle lamellae, while older vacuolated cells degenerate. This results into long cell aggregates separated by large intercellular spaces, giving the callus its friable appearance. The microscopical sections showed a striking difference between friable and compact callus. Vascular elements were not found in the friable callus. On the contrary, vascular bundles were prominent in compact callus. Friable callus is therefore correlated with a less differentiated state than compact callus. The embryogenic potential of friable callus is situated in embryogenic units. These are aggregates of small isodiametric cells containing a central nucleus, an electron-dense cytoplasm, and many organelles. Proliferation was only observed in these cells, which are therefore presumed to generate new embryogenic units, somatic embryos, and vacuolated callus cells. The results further indicate that discrete masses of embryogenic cells, possibly early embryoids, have a unicellular origin. Key words: in vitro culture, callus, somatic embryogenesis, ultrastructure, Zea mays.
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23

WIJAYA, BARAELA EZRA, RINDANG DWIYANI, and NI NYOMAN ARI MAYADEWI. "Induksi dan Multiplikasi Kalus Eucalyptus sp. pada Berbagai Media Callus Induction Medium (CIM) Secara In Vitro." Agrotrop : Journal on Agriculture Science 13, no. 1 (January 17, 2023): 76. http://dx.doi.org/10.24843/ajoas.2023.v13.i01.p07.

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Callus Induction and Multiplication of Eucalyptus sp. on Various Media Callus Induction Medium (CIM) In Vitro. Callus induction in vitro can be done by planting specific part of plant such as leaf, root, stem, etc in aseptic condition by using medium that contains 2,4-D, a plat regulator for promoting callus formation. One of the objectives of callus production in vitro is to form secondary metabolites, because eucalyptus is known as a producer of essential oils. This research is an initial study for in vitro production of secondary metabolites for further research. The aim of this study was to determine the most suitable callus induction medium for callus formation and callus multiplication for Eucalyptus grown in vitro. This research was designed as a Completely Randomized Design (CRD), five treatments, repeated five times. Woody Plant Medium (WPM) was used as the basic media. Several Callus induction Medium (CIM) were used as treatments, i.e. CIM1 = 1 ppm 2,4-D; CIM2 = 1,5 ppm 2,4-D; CIM3 = 2 ppm 2,4-D; CIM4 = 1 ppm NAA+1 ppm BAP; and CIM5 = 2 ppm NAA+ 2 ppm BAP. Variables observed were the time of curly leaf formation, time of the emerging callus, colour and texture of the callus, callus’s weight, callus surface area, the weight of callus after multiplication. The result showed that CIM4 and CIM5 had the fastest emerging callus (7 days after planting), CIM1 and CIM2 formed green and crumb texture of callus, CIM1 produced the highest of callus’s weight, and CIM2 resulted in the highest of callus’s surface area. In conclusion, the most suitable CIM for callus induction was CIM1, while for callus multiplication was CM2.
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24

Hidayat, Rahmad, Abdullah Abdullah, and Aminah Aminah. "PEMANTAPAN MASA INKUBASI KALUS KEDELAI SECARA IN VITRO DALAM MEDIA MS (Murashige dan Skoog) B5 (Gamborg)." AGrotekMAS Jurnal Indonesia: Jurnal Ilmu Peranian 2, no. 2 (May 18, 2022): 60–70. http://dx.doi.org/10.33096/agrotekmas.v2i2.192.

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Combination of Soybean Callus Incubation Period in vitro in MS (Murashige and Skoog) and B5 (Gamborg) Media (Supervised by Abdullah and Aminah). This study aimed to find a suitable medium for stabilizing the incubation period of soybean callus in vitro for the purposes of further selection. This research was conducted at the Laboratory of Bio-Science and Plant Reproduction Biotechnology, Agronomy Department, Agriculture Faculty, Hasanuddin University, Makassar, South Sulawesi. This research was done from June 2020 to October 2020. This research was conducted using a Completely Randomized Design (CRD) with two-factor treatment. The first factor was the type of medium (M1) with 2 levels of treatment. The second factor was the callus incubation period (W) with 5 levels of treatment. The data were analyzed using analysis of variance and 5% LSD. The parameters observed were callus wet weight, callus color and callus structure. The results showed that the MS subculture medium in stabilizing the incubation period of soybean callus tended to be better to callus color and callus structure. The highest wet weight of callus was in the M1W2 treatment with an average of 1.40 callus color scoring values, namely, 5.8 in the white color category and the callus structure with a scoring value of 4, namely callus which had a crumb texture (easily separated). Callus weight tends to be better on B5 medium. The incubation period for soybean callus tended to be better within 14 days, after subculturing on all types of medium (MS and B5) and subsequently decreased by increasing incubation time. The type of incubation medium did not interact with the callus incubation period.
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25

Tan, Yanping, Wen Hu, Xin Xu, Jie Zhou, Chuntai Wang, Xuequn Liu, and Gang Cheng. "Polyamine Plays a Role in Subculture Growth of in Vitro Callus of Indica Rice." Acta Biologica Cracoviensia s. Botanica 59, no. 1 (June 1, 2017): 105–12. http://dx.doi.org/10.1515/abcsb-2017-0001.

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AbstractIn vitroembryogenic callus is a critical factor for genetic transformation of rice, especially for indica varieties. In this study, we investigated the relationship between polyamines, including putrescine (Put), spermidine (Spd) and spermine (Spm), and callus browning, and we studied the effect of exogenous Put on callus regeneration and on the content of endogenous polyamines. In addition, the expression levels of arginine decarboxylase gene (Adc1) and S-adenosylmethionine decarboxylase gene (Samdc) in embryogenic callus were studied by quantitative Real-time PCR analysis. The results showed that the contents of endogenous Put and Spd in the browning callus were significantly lower than those in normal callus. Exogenous Put could effectively improve the growing state of callus of indica rice and enhance the development of embryogenic callus. The content of endogenous polyamines in embryogenic callus, especially Spd and Spm, was increased after addition of exogenous Put. Additionally, exogenous Put also had an obvious impact on the expression levels ofAdc1but partial effect on the expression levels ofSamdcgene. This study could increase the knowledge of both embryogenic callus induction and polyamine catabolism in callus in indica rice.
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26

Agha, Hasan M., Khairul Adzfa Radzun, Norrizah Jafaar Sidik, and Ali H. Jawad. "Callus Induction of Fenugreek Trigonella Foenum-Graecum via Auxin Combined with Cytokinins Hormones, and Assessment of Toxicity via Brine Shrimp Assay." Journal of Asian Scientific Research 12, no. 1 (March 25, 2022): 12–27. http://dx.doi.org/10.55493/5003.v12i1.4449.

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Trigonella foenum-graecum (TFG) is a significant leguminous plant with diverse pharmacological effects. However, the resistant character of this plant accounts for significant difficulties in vitro multiplication, justifying the necessity to try new techniques for in vitro propagation of this plant. Hence, this study reports the effects of BAP, NAA, and 2,4-D on in vitro callus formation of seeds. The toxicity properties of seeds, plant, and callus aqueous extracts of TFG were measured by brine shrimp assay (BSA). Callus index, frequency of callus, callus weight, and morphology of callus were recorded after 30 days of culture. No callus formation was observed in the absence of plant growth regulators. The maximum callus formation observed in the MS media containing 1.0 mg/l 2,4-D, the highest mean of the callus index (52±9.5) with 100% frequency and callus yield (0.52±0.08 g) in 30 days of culture. The highest mean of callus index (37±0 4.05) for combination hormones with 100% callusing and yield (0.37±0.02 g) in 30 days of culture by 1.0 mg/l BAA with 0.5 mg/l NAA. Seeds extract of TFG showed the highest toxicity (954.99 µg/ml), aqueous plant extract (1237.98 µg/ml), and aqueous callus extract (1801 µg/ml) from BSA. Comparing individual hormones, the highest amount of callus in TFG can be yielded 2,4-D hormone alone, and a combination of BAP and NAA can yield 100% callus.
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27

Habibah, Noor Aini, Ananda Lutfiah, Alin Liana, Woro Anindito Sri Tunjung, Meti Indrowati, and Furzani Pa’ee. "Callogenesis of Dayak Onion (Eleutherine palmifolia) Bulb in response of Picloram, 2,4-D, and Kinetin." Biosaintifika: Journal of Biology & Biology Education 15, no. 2 (August 15, 2023): 270–80. http://dx.doi.org/10.15294/biosaintifika.v15i2.46501.

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Dayak onion contains bioactive compounds that are traditionally used for medicine. The production of bioactive compounds can be increased through callus culture. This study aims to analyze the effect of the combination of plant growth regulator on the growth of Dayak bulb callus. The design of this study used a completely randomized factorial design with 2 factors. The first factor is the concentration of auxin. While the second factor is the concentration of kinetin. The parameters observed include the percentage of callus, the time of callus formation, fresh weight, dry weight, and callus morphology. In this study, a callus growth curve was also made to determine the best harvest time. The results showed that highest callus fresh weight was obtained in the P3K1 treatment. The percentage of callus formation in the picloram + kinetin treatment was higher than in the 2,4-D + kinetin treatment. The time of callus formation occurred in the combination of picloram + kinetin faster than the 2.4-D + kinetin treatment. The callus color is diverse, while the texture of the entire callus is compact. The best callus induction medium for Dayak onion bulbs is to use picloram 2-4 ppm + kinetin0.025-0.5 ppm. The results of this study provide optimal growth regulatory information for the induction of dayak onion callus that has never been reported before. This information can be the basis for the development of methods of production of bioactive compounds from dayak onions through callus culture.
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28

Mackay, Wayne A., Timothy J. Ng, and Freddi A. Hammerschlag. "Cucumis melo L. Callus Response to Toxins Produced by Myrothecium roridum Tode ex. Fries." Journal of the American Society for Horticultural Science 119, no. 2 (March 1994): 356–60. http://dx.doi.org/10.21273/jashs.119.2.356.

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Studies examining exposure methods and callus type were conducted to develop an in vitro selection system using roridin E as a selection agent. Vacuum infiltration of callus with the toxin solution was the only successful selection method at the concentrations tested. Primary callus (callus originating directly from the explant) was not sensitive to roridin A or E at the concentrations used. Secondary callus (callus produced from primary callus) exhibited a differential response to roridins A and E similar to that of detached-leaf assays. Electrolyte leakage studies of callus were not conclusive in establishing the membrane as the site of toxin action or useful for screening tolerance in vitro. A small percentage of callus from tolerant and susceptible cultivars survived repeated exposure to roridin E at 50 μg·ml-1.
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29

Muralitharan, MS, SF Chandler, and RFMV Steveninck. "Physiological Adaptation to High Ion Concentrations or Water Deficit by Callus Cultures of Highbush Blueberry, Vaccinium corymbosum." Functional Plant Biology 20, no. 2 (1993): 159. http://dx.doi.org/10.1071/pp9930159.

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Callus cultures of highbush blueberry were selected for 10 passages on medium supplemented with 50 rnol m-3 NaCl, 50 rnol m-3 KCl, 25 mol m-3 Na2SO4, 25 mol m-3 K2SO4 or 100 mol m-3 mannitol. On all salts, growth of selected callus was greater (200-250%) than that of non-selected callus, and selected callus grew optimally on the type of salt on which it was selected. Conventional (whole plant analysis) and electron probe X-ray microanalysis showed that selected callus accumulated more ions (approximately 1.5-3.0-fold) than non-selected callus on all salts, and there was a positive correlation between vacuolar ion concentration and fresh weight. Growth of NaCl-selected callus but not non-selected callus was greatly enhanced (2.25-fold) in the presence of 100 mol m-3 mannitol, while growth of a mannitol-selected callus line was also enhanced. In callus grown on NaCl or mannitol, slight increases in levels of glycinebetaine, choline and proline were measured. Sucrose, glucose, fructose, sorbitol and malate concentrations significantly increased in callus grown on NaCl or mannitol, and selected callus produced 4-fold more sugars than non-selected callus. The total increases in concentrations of all measured sugars were 210 μmol gFW-1 in NaCl-selected callus grown on 50 mol m-3 NaCl, and 296 μmol gFW-1 in mannitol selected callus grown on 100 rnol m-3 mannitol. The results of this study indicate that adaptation of blueberry callus cultures for optimal growth on salt-containing media is probably due to adaptation to water stress, not tolerance to specific ions. Osmotic adjustment, achieved by ion uptake and production of sugars, appears to be the physiological mechanism of adaptation.
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30

Yang, Yali, Peiting He, Jiayi Zhang, Lijian Jin, Baoying Chen, Xiaoxing Lao, Shengyuan Zhang, Lubin Zhang, Ming Zhai, and Ying Liu. "Induction of callus in Cassia mimosoides." E3S Web of Conferences 189 (2020): 02017. http://dx.doi.org/10.1051/e3sconf/202018902017.

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In this study, the young leaves of Cassia mimosoides were used as explants. On the basis of 1/2 MS +2.0 mg/L 6-BA +0.5 mg/L NAA medium, we observed the effect of adding different factors on the callus induction of C. mimosoides. The results shown that after 20 days of callus induction, it was found that the addition of 150 mg/L vitamin C (VC) did not affect the induction efficiency, but the growth of callus became slower; and when the concentration was higher than 150 mg/L, the formation of callus would be inhibited. The induction of callus would be restrained while adding different concentrations of Cu2+ into mediums. Moreover, glutamine (Gln) had little effect on the induction of callus. Furthermore, the addition of hydrolyzed casein (CH) would not affect the formation of callus, but the high concentration of CH could affect the growth status of callus. The induction efficiency of callus was severely inhibited by exogenous addition of silver nitrate (AgNO3), and the growth status of callus was poor, and the phenomenons of early rooting had happened at the same time. The results of this study provided a theoretical basis for the subsequent optimization of bud proliferation and rooting in C. mimosoides.
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Schneider, K. E., D. Speranzini, and A. R. Biggs. "Ontogeny of shoot regenerants on excised immature peach embryos." Canadian Journal of Plant Science 72, no. 2 (April 1, 1992): 497–506. http://dx.doi.org/10.4141/cjps92-063.

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Excised immature peach embryos when cultured on an auxin-containing medium produced both a friable callus and a nodular callus. Upon further subculturing on an auxin-reduced medium only the nodular callus differentiated into highly regenerative nodular callus. This callus comprised dense meristematic cells which arose near or at the surface of the nodular tissue. Cells containing starch granules often surrounded these meristemoid areas. Shoots could be induced to form on shoot-inducing medium from the meristemoid areas. Vascular connections developed between the parental material and the shoot. The friable callus never appeared to differentiate into a nodular callus on either friable callus or nodular callus medium, nor were plantlets derived from this tissue on shoot-inducing medium. No stages of somatic embryogenesis were ever apparent. The evidence supports the view that the regenerative shoots were derived from indirect organogenesis on nodular callus derived from the embryonic tissue.Key words: Organogenesis, Prunus persica (L.) Batsch, shoot formation
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MATSUO, Masaki, Toshitaka UCHINO, and Hidemitsu MASUDA. "Effect of Electric Current Flow to Callus on the Multiplication of Asparagus Callus." Shokubutsu Kojo Gakkaishi 4, no. 1 (1992): 10–14. http://dx.doi.org/10.2525/jshita.4.10.

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Adkins, SW. "Cereal Callus Cultures: Control of Headspace Gases Can Optimise the Conditions for Callus Proliferation." Australian Journal of Botany 40, no. 6 (1992): 737. http://dx.doi.org/10.1071/bt9920737.

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The protective conditions under which callus cultures are grown to prevent microbial contamination and to reduce tissue desiccation cause the accumulation of volatiles in the vessel headspace and reduce the availability of oxygen for respiration. To demonstrate the importance of the gaseous atmosphere to culture growth a study was undertaken on non-morphogenic rice and wheat callus incubated under a number of environmental conditions. Changes in the gaseous atmosphere above rice (Oryza sativa L.) callus during routine culture in a petri dish suppressed growth and promoted necrosis. Incubating callus under a continuous flow of gas mixtures of known composition suggested that the inhibition of growth was caused by the accumulation of high levels of ethylene and to the rapid depletion of oxygen. In order to evaluate the importance of ethylene accumulation aminoethoxyvinyl glycine (AVG), I-aminocyclopropane-I-carboxylic acid (ACC) and silver nitrate (AgNO3) were added to the nutrient medium and ethylene was measured during callus culture. Ethylene restricted callus growth particularly under high (35°C) compared with moderate (25°C) incubation temperatures and under illuminated compared with dark incubation. Under illuminated incubation at 25°C, AVG ( 5 μM ) and AgNO3 (50 μM) improved rice callus growth by 69 and 54% respectively while ACC (100 μM) decreased growth by 15%. Furthermore, rice callus growth was better in large compared with small culture vessels since ethylene accumulation was reduced. In contrast, wheat (Triticum aestivum L.) callus grew well in the petri dish system and released very little ethylene into the culture vessel headspace. Growth was better under illuminated than darkened conditions and under moderate (25°C) compared with high (35°C) incubation temperatures. Furthermore, wheat callus growth was only marginally better in large compared with small culture vessels. Ethylene was not a restrictive factor of wheat callus growth since only low levels were detected in all conditions of incubation. Better control of ethylene and increased oxygen availability could be a way of increasing cell and tissue production for genetic engineering studies of otherwise recalcitrant species such as rice, and may be a way of improving manipulation of wheat.
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Xiang, Bei Bei, Ye Rong Zhu, Wen Juan Wang, Yan Ling Bai, and Yong Wang. "Improvement of Vindoline Production in a Catharanthus roseus callus Line." Advanced Materials Research 345 (September 2011): 375–80. http://dx.doi.org/10.4028/www.scientific.net/amr.345.375.

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The influence of polyploidity on the accumulation of vindoline in Catharanthus roseus callus cultures was investigated. The callus line (T1) was induced from a tetraploid leaf. Total alkaloids in the callus were extracted and analyzed by LC-MS for qualitification and HPLC for quantification. Results showed that T1 callus cultures could accumulate vindoline to a high level. The highest accumulation of vindoline found in the callus was 0.11 mg g−1 DW. Our results demonstrate that polyploidity could influence the chlorophyll content or chloroplast development and improve vindoline biosynthesis in callus cultures. The T1 callus cultures also accumulated a substance which was not present in the diploid callus. The substance was preliminarily identified by LC-MS as deacetylvindoline, the direct precursor of vindoline biosynthesis. T1 callus could be used for genetic manipulation of the biosynthesis of vinblastine and vincristine, the two important antitumor drugs, and therefore, have potential commercial value.
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Royani, J. I., H. Khairiyah, F. R. Mira, Karyanti, and N. Watanabe. "The effect of callus induction media for somatic embryo formation in Hevea brasiliensis Muell. Arg." IOP Conference Series: Earth and Environmental Science 1160, no. 1 (April 1, 2023): 012007. http://dx.doi.org/10.1088/1755-1315/1160/1/012007.

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Abstract The method for micro-propagation of rubber (Hevea brasiliensis Muell. Arg) by somatic embryo has been conducted. The aim of the research will describe the effect of media used for callus induction on the somatic embryo formation of rubber plants. Embryonic callus had been induction in 3 media, i.e.; MH1, MH2, and MH3. After callus induction, all callus were induced in media for embryo somatic expression. Maturation of cotyledons was performed using liquid media with filter paper as a bridge to absorb the media. The results showed that the percentage of callus was induced differently for each media. MH3 media was the best media for callus induction with 62.5% of callus forms, but for embryonic callus, MH1 was the best media. Not all of the calluses could be expressed to become somatic embryos. Most calluses derived from MH1 media were the best callus for somatic embryos formed.
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Anjani, Dianita Dewi, and Ratnawati. "The Effect of BAP and NAA Combination on Callus Induction of Aglaonema Siam Aurora Leaf Explants in Vitro." Indonesian Journal of Bioscience (IJOBI) 1, no. 2 (October 31, 2023): 85–92. http://dx.doi.org/10.21831/ijobi.v1i2.213.

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This research aims to determine the effect of the Naphthalene Acetic Acid and Benzyl Amino Purine combination on callus growth of Aglaonema Siam Aurora leaf explants and to determine the combination level at the most optimal concentration on the callus growth of Aglaonema Siam Aurora leaf explants. Observation variables included callus emergence time, explants percentage of formed callus, callus length, and explant life percentage. The explants that where used were the first young leaves explants from shoots that were opened, with 2x2 cut size. Results show that the addition of a NAA and BAP combination had an effect on . Combination of 1.2 ppm BAP + 1 ppm NAA treatment is the most optimum treatment for callus growth with a callus percentage of 87.5%, the callus start time at week 3, the average callus length is 0.77 mm and the percentage number of survived explants is 100%
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Yang, Lin, Jinge Sun, Congyu Yan, Junyi Wu, Yaya Wang, Qiuting Ren, Shen Wang, Xu Ma, Ling Zhao, and Jinsheng Sun. "Regeneration of duckweed (Lemna turonifera) involves genetic molecular regulation and cyclohexane release." PLOS ONE 17, no. 1 (January 6, 2022): e0254265. http://dx.doi.org/10.1371/journal.pone.0254265.

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Plant regeneration is important for vegetative propagation, detoxification and the obtain of transgenic plant. We found that duckweed regeneration could be enhanced by regenerating callus. However, very little is known about the molecular mechanism and the release of volatile organic compounds (VOCs). To gain a global view of genes differently expression profiles in callus and regenerating callus, genetic transcript regulation has been studied. Auxin related genes have been significantly down-regulated in regenerating callus. Cytokinin signal pathway genes have been up-regulated in regenerating callus. This result suggests the modify of auxin and cytokinin balance determines the regenerating callus. Volatile organic compounds release has been analysised by gas chromatography/ mass spectrum during the stage of plant regeneration, and 11 kinds of unique volatile organic compounds in the regenerating callus were increased. Cyclohexane treatment enhanced duckweed regeneration by initiating root. Moreover, Auxin signal pathway genes were down-regulated in callus treated by cyclohexane. All together, these results indicated that cyclohexane released by regenerating callus promoted duckweed regeneration. Our results provide novel mechanistic insights into how regenerating callus promotes regeneration.
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Huda, Moch Faizul, and Serafinah Indriyani. "Effect of Benzyl Adenine Concentration on Callus Induction of Geranium Plants (Pelargonium graveolens L'Her) from Petiole and Leaf Explants." Journal of Experimental Life Sciences 10, no. 1 (June 30, 2020): 20–23. http://dx.doi.org/10.21776/ub.jels.2019.010.01.04.

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Geranium plant (Pelargonium graveolens L'Her) is one of the geranium oil producing plants that has many benefits. Callus culture is a technique that can be used to plant multiplication and increase production of secondary metabolites. This study aims to determine the effect of the concentration of Benzyl Adenine on the formation of geranium callus from petiole and leaf explants. Callus induction was carried out by culturing petiole and leaf explants on MS medium + 0.1 mg.L-1 NAA + Benzyl Adenine (0; 0.5; 1; 1.5 and 2 mg.L-1). Callus morphological parameters, percentage of callus formation, and time of first callus formation were observed. The formation of geranium callus influenced by the explant type and the concentration of Benzyl Adenine. In the 2nd week, the geranium callus was initiated, light green colored with a compact callus texture. At 4th week, the percentage of callus formation containing NAA 0.1 mg.L-1 of petiole and leaf explants was 20% and 8%, whereas the percentage of callus formation on medium containing 0.1 mg.L-1 NAA combined with 0.5-2 mg.L-1 Benzyl Adenine of petiole and leaf explants was 52-80% and 24-52%. The best percentage of callus formation was found on the culture medium containing 1 mg.L-1 Benzyl Adenine, equaled 80% of petiole explants, and 52% of leaf explants.
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Revutskaya, A. Z., A. V. Holubenko, N. V. Nuzhyna, H. O. Rudik, and N. Yu Taran. "Introduction to in vitro culture and callus initiation in Salvia hispanica L. (chia)." Visnik ukrains'kogo tovaristva genetikiv i selekcioneriv 17, no. 1 (July 31, 2019): 33–37. http://dx.doi.org/10.7124/visnyk.utgis.17.1.1198.

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Aim. Preparation of aseptic seedlings Salvia hispanica L., callus initiation in vitro and establishment of primary explants suitable for the callus production. Methods. Seeds are sprouted on our own modification of conventional methods. The non-hormonal Murashige-Skoog agarized nutrient medium was used as basic medium for the experiments. Parts of one-month seedlings (roots, hypocotyl, cotyledon leaves) were used as explants for the use of the colza. We added growth regulators (BAP, 2,4-D) in different concentration combinations into the nutrient medium for callus initiation. Statistical processing was performed in Microsoft Office Excel. Results. Aseptic S. hispanica seedlings have been obtained. The callus growth was initiated on all types of explants, the dependence of the callus intensity on the type of explants and the growth regulators content in the nutrient medium was established. Morphogenic callus and root-regenerants have been obtained. Conclusions. Hypocotyl was the most suitable primary explant for callus growth. Seedlings, leaves and roots showed low morphogenetic capacity. The nutrient medium with an elevated 2,4-D content was the most effective for initiation of callus genesis and proliferation of non-morphogenous callus. A high concentration of 2,4-D in the medium improves S. hispanica callus growth but suppresses its morphogenic ability.Keywords: Salvia hispanica (Chia), in vitro culture, callus.
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Galibina, N. A., M. A. Ershova, R. V. Ignatenko, K. M. Nikerova, I. N. Sofronova, and M. N. Borodina. "Cytogenetic and Biochemical Characteristics of Callus <i>Pinus sylvestris</i> L." Физиология растений 70, no. 1 (January 1, 2023): 100–112. http://dx.doi.org/10.31857/s0015330322100244.

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A comprehensive assessment was carried out of the changes occurring during the darkening of the callus obtained from vegetative buds of 40-year-old Scots pine treesPinus sylvestrisL. Based on biometric assessment of callus (intensity of callus formation, proportion of light callus, callus mass) from 32 analyzed trees (16 genotypes represented by two clones), two genotypes with high callus-forming ability were singled out. Analysis of mitosis showed that, although the proportion of aberrant cells in the callus does not exceed the rate of spontaneous mutation forP. sylvestris, the range of violations at the stage of meta-, ana-, and telophase in the callus culture was wider compared to that in the seed progeny of the same pine trees. Darkening of the callus was accompanied by a decrease in sucrose metabolism in the cell (decrease in cytoplasmic, vacuolar invertase and sucrose synthase) and a significant decrease in peroxidase activity. At the same time, the activity of apoplast invertase was maintained at a constant level. The activity of superoxide dismutase, catalase, polyphenol oxidase, and phenylalanine ammonia lyase, on the contrary, was higher in dark callus. The possible use of the studied enzymes as biochemical markers of the transition to darkening callus pine crops is discussed.
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Setiaji, Arkan, RR Rifka Annisa, Rumiyati Rumiyati, and Endang Semiarti. "Induction and Growth Kinetics Callus of Tomato (Solanum lycopersicum)." Biosaintifika: Journal of Biology & Biology Education 12, no. 1 (April 23, 2020): 35–41. http://dx.doi.org/10.15294/biosaintifika.v12i1.21704.

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Plant callus extracts are potential to be developed as ingredient in skincare products. Tomato callus is supposed to contain protein-derivatives and or other components such as secondary metabolites that play a role in skin regeneration. Therefore the production of calli is important to be studied for callus sustainable supply. This research aims to obtain optimum medium for callus induction and to analyze tomato callus development anatomically. In vitro culture response was assessed in tomato plant (Solanum lycopersicum L. ‘Permata’) for optimum callus induction. Seeds were grown on ¼ MS medium for 10-15 days. Hypocotyl was excised and cultured on MS medium + 2 mg/l 2,4-D for 15 days as the explants for callus induction. Callus was transferred to MS medium with 8 variations of PGRs including the combination of BAP + NAA, and 2,4-D. Both fresh and dry weight was measured every 5 days over 60 days to establish the growth kinetics and growth efficiency of callus. Anatomic characters of calli were examined through paraffin-embedded method. The result showed of MS medium supplemented with 2.0 mg/l NAA and 0.2 mg/l BAP is optimum for tomato callus induction, based on highest number of the absolute growth rate on fresh weight (73.77% per day), dry weight (3.84% per day), and callus initiation time (5.56 days) achieved by the medium. Cells in the ground tissue of tomato hypocotyl are competent to be dedifferentiated into a callus. This research results were expected to find out suitable methods for tomato callus production in preparation for skincare uses.
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Özgen, Murat, Melahat A. Birsin, and Berk Benlioglu. "Biotechnological characterization of a diverse set of wheat progenitors (Aegilops sp. and Triticum sp.) using callus culture parameters." Plant Genetic Resources 15, no. 1 (August 14, 2015): 45–50. http://dx.doi.org/10.1017/s1479262115000350.

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It is known that genetic diversity is the most important factor in classical and modern plant breeding. The considerable increase in the number of transgenic crops reveals the value of new plant genetic resources. In this study, a set of 12 wheat progenitors were screened for tissue culture parameters such as callus induction, callus weight, regeneration capacity of callus and callus efficiency using mature embryos. Embryos were excised from imbibed seeds of the progenitors. The excised embryos were placed scutellum upwards in dishes containing 2 mg/l 2,4-dichlorophenoxyacetic acid (2,4-D) for callus induction. The developed calli and regenerated plants were maintained on 2,4-D free MS medium. When mature embryos of 12 wheat progenitors (Aegilops sp. and Triticum sp.) were compared, significant differences were detected in callus induction frequency, weight of callus, regeneration capacity and culture efficiency. A significant genotypic effect was observed on the culture responses. Of the 12 wheat progenitors tested, Aegilops umbellulata had the highest regeneration capacity of callus. Aegilops biuncialis created the most regenerable calli because of the highest callus induction and culture efficiency. In the experiment, callus induction was significantly correlated with callus weight (r= 0.820) and regeneration capacity (r= 0.955). Weight of callus was significantly correlated with regeneration capacity (r= 0.740), while there was no significant correlation between callus induction frequency and culture efficiency (r= 0.350). Our results showed that, generally, mature embryos of some Aegilops and Triticum species have a high regeneration capacity, and therefore, can be used as an effective explant source for the successful application of biotechnology in crop improvement.
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HARDIYANTO, ARIF, SOLICHATUN SOLICHATUN, and WIDYA MUDYANTINI. "The effect of varied NAA concentration agains growth and flavonoid content of Gynura procumbens (Lour) Merr. callus." Biofarmasi Journal of Natural Product Biochemistry 2, no. 2 (August 2, 2004): 69–74. http://dx.doi.org/10.13057/biofar/f020205.

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The aim of this research is to study the influence of naphthalene acetic acid (NAA) concentration variation in the callus growth and flavonoid content of daun dewa [Gynura procumbens (Lour) Merr.]. The research framework relied on the benefit of daun dewa as anti cancer drug. This matter enabled by its compound cytotoxic from compound flavonoid contained. Culture in vitro technique serves the purpose of medium to produce the secondary metabolism. Addition of NAA as treatment for the induction of callus will promote cell enlargement and the growth of callus so that callus which later will be analysed by its flavonoid content. This research was conducted with two phase. First phase represented the phase of antecedent attempt in the form of callus induction to get the good callus. Addition of NAA and kinetin conducted to promote the cell proliferation. The parameter of this phase is texture, colour, and moment of callus appearance. Second phase is the form of NAA concentration variation treatment phase. At this research, it conducted with the random device complete with one factor that is 5 concentration NAA is 0 mg/L; 0,5 mg/L; 1,0 mg/L; 1,5 mg/L and 2,0 mg/L. The parameter of this phase is wet weight of the callus, dry weight of the callus, texture of the callus, colour of the callus, and flavonoid content. The result of this research indicated that the addition NAA does not influence on callus growth and the flavonoid content of callus daun dewa.
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IRMAWATI, IRMAWATI, SOLICHATUN SOLICHATUN, and ENDANG ANGGARWULAN. "The growth and reserpine content of callus culture of Rauvolfia verticillata on the variation of sucrose concentration in MS medium." Biofarmasi Journal of Natural Product Biochemistry 5, no. 1 (February 17, 2007): 38–46. http://dx.doi.org/10.13057/biofar/f050105.

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Irmawati, Solichatun, Anggarwulan E. 2007. The growth and reserpine content of callus culture of Rauvolfia verticillata on the variation of sucrose concentration in MS medium. Biofarmasi 5: 38-46. The aim of this research was to study the effect of the variation of sucrose concentration on the growth and reserpine content on callus culture of Rauvolfia verticillata (Lour.) Baillon. The research was conducted with callus culture method consisted of two stages. The first stage was callus initiation to induce callus from leaf explant of R. verticillata, and the second stage was the reserpine production on treatment medium. This research used a Completely Randomized Design (CRD) by one factor, i.e. the variation of sucrose concentration. The sucrose concentration consisted of five levels, i.e. 0 g/L, 10 g/L, 20 g/L, 30 g/L and 40 g/L, each concentration in five replicates. The collected data consisted of qualitative and quantitative data. The qualitative data, callus morphology, was presented descriptively. The quantitative data, included fresh weight callus, dry weight callus and reserpine content, were analyzed by using ANOVA and followed by DMRT at 5% significance level. The result of research showed that the variation of sucrose concentration influenced fresh weight callus, dry weight callus and reserpine content. The increasing of sucrose concentration tended to raise callus growth, which could be seen from the fresh and dry weight callus. The highest fresh weight callus was found in medium with sucrose concentration of 20 g/L, while the highest dry weight callus was found in medium with sucrose concentration of 40 g/L. The increasing of sucrose concentration until 30 g/L raised reserpine content, but the sucrose concentration over 30 g/L decreased the reserpine content.
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TRIMULYONO, GUNTUR, SOLICHATUN SOLICHATUN, and SOERYA DEWI MARLIANA. "Callus growth and essential oil of nilam (Pogostemon cablin (Blanco) Bth.) on the treatment with -naphtalen acetic acid and kinetin." Biofarmasi Journal of Natural Product Biochemistry 2, no. 1 (February 2, 2004): 9–14. http://dx.doi.org/10.13057/biofar/f020102.

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The objectives of the research were to study the effect of adding of α - naphtalen acetic acid (NAA) and kinetin on callus growth and essensial oil production from nilam (Pogostemon cablin (Blanco) Bth.) callus culture. The outline of the research was the callus growth and secondary metabolite production from plant’s body could be triggered by the occurrence of plant growth regulation. The addition NAA and kinetin in culture’s medium would influence cell proliferation and synthesis of protein, so that both can induce callus growth and secondary metabolism production from the cell that have been cultured. According to the research objectives, the research was done by using in vitro callus culture method to obtain callus from explant P. cablin leaf and to induce essensial oil production. In vitro culture process consist of two stages. First stage was the callus initiation medium induced the callus from explant. The experiment was done by medium Murashige-Skoog (MS) with 2,4-D 0,5 mg/l and kinetin 0.5 mg/l; and the second stage was the medium treatment induced callus growth and essensial oil production. The research used factorial completely randomized design with two factors (NAA concentration: 0mg/l, 0,5 mg/l, 1,0 mg/l, 2,0 mg/l; and kinetin concentration: 0 mg/l, 0,5 mg/l, 1,0 mg/l, 2,0 mg/l) with 3 replicates. The collected data were qualitative data (callus morphology include texture and colour callus) and quantitative data (callus growth rate, callus dry weight and essensial oil content). Data were analyzed by Anova and followed by DMRT 5% confidence level and correlation regretion. The result of the research indicated the treatment with addition plant growth regulation (NAA and kinetin) on Murashige-Skoog’s medium had significant effect on callus growth rate but it didn’t have significant effect on callus dry weight and the increase of produced essential oil.
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Arieswari, Ni Nyoman Nila, Ida Ayu Astarini, Junita Hardini, Austin Ryan Garrido, Debora Margareth, Jennifer Crismonika, and Sebastian S. Cocioba. "Callus Induction In Leucaena (Leucaena leucocephala (Lam.) de Wit) As An Effort To Provide Target Transformation Through Agrobacterium tumefaciens." International Journal of Biosciences and Biotechnology 10, no. 1 (October 1, 2022): 13. http://dx.doi.org/10.24843/ijbb.2022.v10.i01.p02.

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Leucaena is a plant that produces biomass productivity in the form of hardwood for fuel with low humidity and high calorific value. However, Leucaena is also classified as an invasive plant which can cause the urgency of native plant species and ecosystems in Indonesia. Therefore, the formation of sterile Leucaena needs to be done, one of which is through genetic transformation using Agrobacterium tumefaciens. Callus is used as a target for transformants in the genetic transformation process, so it is necessary to use appropriate media and PGR. This study aimed to determine the type of media and the concentration of 2,4-D on callus induction. This research is an experimental study with a completely randomized design (CRD) method with two factors. The first factor is the type of media (MS and WPM) and the second factor is the concentration of 2,4-D (0; 0.25; 0.50; 0.75; 1.00; 1.25 and 1.50 mgL-1 ). Each treatment was repeated three times so that 42 experimental units were obtained. Parameters observed were callus initiation, callus fresh weight (gram), callus texture and color. Quantitative data is analyzed by analysis of variance (ANOVA). The results showed that the use of media had a significant effect (P<0.05) on callus fresh weight. The use of 2,4- D concentration had a significant effect (P>0.05) on callus texture. The use of WPM media resulted in the fastest callus emergence time (6.67±0.57), the best callus texture (crumb callus type 2) and the best callus color (green). Meanwhile, the highest fresh weight (2,48±0.83) was in the use of MS media. The fastest callus emergence time occurred in the control (without the addition of 2,4-D) (7.33±0.57 and 6.67±0.57), the highest average fresh callus weight (2,48±0.83 and 2.35±0.32) occurred in the treatment with the addition of 1.00 mgL-1 2,4-D with a crumb callus texture of type 2 and callus green color only appeared in the treatment with a concentration of 0.25 mgL-1.
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Rathod, Zankhana, Santoshkumar Singh, and O. P. Saxena. "BIOCHEMICAL PROFILE OF EMBRYONIC AND NON-EMBRYONIC CALLUS OF CLERODENDRUM PHLOMIDIS L." International Association of Biologicals and Computational Digest 1, no. 2 (October 6, 2022): 1–5. http://dx.doi.org/10.56588/iabcd.v1i2.33.

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Clerodendrum phlomidis L. (Arani) is a medicinal plant used in the treatment of Diabetes mellitus. Enzymatic studies helps to differentiate between embryonic and non-embryonic callus at very early stage. In vitro experiments were conducted using leaf, stem, node, internode, apical bud as an explant. Callus cultured on MS basal medium supplemented with various combinations of 2, 4-D and Kinetin (1 : 1, 2 : 2, 3 : 3, 4 : 4, 5 : 5 mg/l 2,4-D : Kinetin). Large variations were observed in the callus cultures owing to hormonal variations. Higher hormonal concentration 5 mg/l 2, 4-D + 5 mg/l kinetin resulted in somatic embryoids while lower concentrations 1:1, 2:2, 3:3 mg/l, 2,4-D : kinetin resulted in non-embryonic cells. Enzymatic activities of peroxidase, IAA- oxidase, invertase, protease,  and  amylase, polyphenol oxidase (PPO), catalase and enzyme protein were estimated from cytoplasmic as well as wall-bound fractions of 2, 4, 6and 8 week old callus using standard methods. In two week old culture peroxidase and polyphenol oxidase (both wall bound and cytoplasmic) activity increases in non-embryonic callus and decreases in embryonic callus while IAA oxidase and Protease (both wall bound and cytoplasmic) activity decreases in non-embryonic callus and increases in embryonic callus. Total amylase and α-amylase activity of wall bound enzymes increases in non-embryonic callus and decreases in embryonic callus while cytoplasmic activity decreases in non-embryonic callus and increases in embryonic callus. On the basis of the above study enzymes could serve as a biochemical marker to determine the embryonic and non-embryonic callus at very early stage.
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Chahredine, Sadek, and Nadia Ykhlef. "Improvement of Callogenesis Ability by Selecting a Better Hormonal Balance in Potato (Solanum tuberosum L)." International Journal for Innovation Education and Research 4, no. 9 (September 30, 2016): 50–60. http://dx.doi.org/10.31686/ijier.vol4.iss9.589.

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Response in tissue culture is highly genotype dependent, significant genotypic diferences in callus initiation response were observed among both potato genotype investigated. The Spunta variety devleops the best calogenesis in all media compared to Kondor variety depending on hormones concentration,there is a range of variations in days needed for callus initiation,percentage of explant that developed callus, its texture, color and the degre of its formation.Our resutls chow that the callus depends on explant type.Sprout explant respond best to callus formation. the amount of callus ranges from 60% to 90 % for Spunta .Callus color after eight weekwas light green or green yelow for both varieties.The higest amount of callus 100% was obtained with the combination (NAAxBAP)(0.5/1, medium M2) and (2/2 ,medium M3) with Kondor bud explant .In media M1(1/0.5 mg/l),M2 (0.5/1 mg/l) and M3 (2/2 mg/l) with sprout explant of Spunta the amount of 80 %was noted,callus in media M1 and M2 produced microtubers,shoots and roots.
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49

Ayuningrum, Kiki, Iman Budisantoso, and Kamsinah Kamsinah. "Respon Pemberian Hormon 2,4-D dan BAP terhadap Pertumbuhan Subkultur Kalus Kedelai (Glycine max (L.) Merrill) secara In Vitro." Biosfera 32, no. 1 (January 10, 2015): 59. http://dx.doi.org/10.20884/1.mib.2015.32.1.296.

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The purpose of this study was to evaluate the response of administration of a combination of 2,4-D and BAP on the growth of soybean callus subculture and determine the combination of 2,4-D and BAP most good for the growth of soybean callus subculture. The study design used completely randomized design (CRD) with a pattern factorials. The factor one e.i 2,4-D consists of 4 levels, namely: 0, 5, 10, and 15 ppm. A factor of 2 e.i BAP consists of 4 levels, namely: 0, 2, 4, dan 6 ppm. Every combination treatment repeated three times. Parameters measured include the percentage is growing callus, type of callus, dry weight and wet weight of soybean callus. The results showed that administration of the hormone 2,4-D and BAP can spur the growth of soybean callus subculture, the combination of BAP 2 ppm and 10 ppm of 2,4-D is the best combination for a percentage of callus and growing callus types, whereas the wet weight and the weight dried callus is not driven by a combination of the hormone 2,4-D and BAP
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50

Webb, D. T., F. Webster, B. S. Flinn, D. R. Roberts, and D. D. Ellis. "Factors influencing the induction of embryogenic and caulogenic callus from embryos of Piceaglauca and P. engelmanii." Canadian Journal of Forest Research 19, no. 10 (October 1, 1989): 1303–8. http://dx.doi.org/10.1139/x89-200.

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Abstract:
Zygotic embryos of Piceaglauca (Moench) Voss from five half-sib seed families and P. engelmanii Parry from one half-sib family, collected on July 13 and 27 and August 24, were cultured in the presence of 2,4-dichlorophenoxyacetic acid, N6-benzyladenine, and sucrose ranging from 0.5 to 4% for the induction of embryogenic callus and the production of stable embryogenic callus lines. Embryogenic callus was induced from all three collections with all seedlots. The July 13 collection was two to four times more embryogenic than the later collections. Embryogenic callus was induced at all sucrose levels, but 4% sucrose was clearly inferior, whereas 1% was best overall. Factors that favored the induction of embryogenic callus also favored the production of stable embryogenic callus lines. There was a 40% decline in callus line establishment compared with embryogenic callus induction and some seed lots failed to yield embryogenic lines from the two later collections. The formation of caulogenic callus was promoted at 3 and 4% sucrose. Embryos from the August 24 collection were more caulogenic than those from the earlier collections.
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