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Journal articles on the topic 'Shoot tips'

Author: Grafiati

Published: 4 June 2021

Last updated: 1 February 2022

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1

Ruan, Yiqin, and Mark H. Brand. "In Vitro Responses of Tissues from Rhododendron Plants With and Without Tissue Proliferation." HortScience 30, no. 4 (July 1995): 873D—873. http://dx.doi.org/10.21273/hortsci.30.4.873d.

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Rhododendron `Montego' shoot cultures initiated from plants with and without tissue proliferation (TP and NTP) served as explant sources for all studies (Note: in vitro TP shoot cultures produce primarily dwarf shoots, some long shoots, and stem tumors). Calli induced from TP leaves and tumors and NTP leaves were cultured on woody plant (WP) medium containing NAA and 2-iP. During the first 4 weeks of culture, calli from NTP leaves had higher relative growth rates than calli from TP leaves or tumors. However, calli from TP leaves and tumors grew faster than calli from NTP leaves for all subculture periods that followed. Shoot tips (5 mm) were excised from TP dwarf shoots, TP long shoots, and NTP shoots and were cultured on WP medium with or without 15 μM 2-iP. Shoot tips from TP dwarf and long shoots multiplied on medium without 2-iP, averaging 18.4 and 1.7 shoots per shoot tip in 12 weeks, respectively. Shoot tips from NTP shoots only multiplied when maintained on 2-iP-containing medium. When placed on 2-iP-containing medium, both types of TP shoot tips produced clusters of callus-like nodules that gave rise to highly tumorized, short shoots or leafy meristems.

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2

Fukai, Seiichi. "Cryopreservation of chrysanthemum shoot tips." Scientia Horticulturae 45, no. 1-2 (December 1990): 167–74. http://dx.doi.org/10.1016/0304-4238(90)90079-t.

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3

Dahniya, M. T., S. K. Hahn, and C. O. Oputa. "Effect of Shoot Removal on Shoot and Root Yields of Sweet Potato." Experimental Agriculture 21, no. 2 (April 1985): 183–86. http://dx.doi.org/10.1017/s0014479700012461.

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SUMMARYExperiments on harvesting sweet potato as a green vegetable and as a root crop are described. Whole shoots yielded 62% more than shoot tips. Similar total shoot yields were harvested whether tip removal was at two, three or four week intervals. Root yield was decreased by 31 to 48% by removing shoot tips, while removing whole shoots led to root yield decreases of 48 to 62%. Harvesting shoots at two week intervals gave 72% reduction in root yield, compared with 50% with four week intervals. There were fewer and smaller tubers as the frequency of shoot harvests increased. There were varietal differences in response to shoot removal For reasonable yields of both shoot tips and tuberous roots harvesting shoot tips at four week intervals is recommended.

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4

Kozak, Danuta, Marzena Parzymies, Alicja Świstowska, Barbara Marcinek, and Bairam Solomon Ismael. "The influence of explants type and orientation on growth and development of Mandevilla sanderi (Hemsl.) Woodson in vitro." Acta Scientiarum Polonorum Hortorum Cultus 18, no. 4 (August 7, 2019): 111–19. http://dx.doi.org/10.24326/asphc.2019.4.10.

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Mandevilla sanderi is an important commercial ornamental pot plant. Traditional vegetative propagation is limited due to the low rate, therefore there is a need to develop an alternative, more efficient method. There is an interest in development of micropropagation technology for the species, as it allows to obtain a lot of offsprings in a relatively short time. The aim of the present work was to estimate an influence of explants type and position on regeneration of Mandevilla sanderi in tissue culture. Four different types of explants (leafy shoot tips, decapitated leafy shoot tips, defoliated shoot tips, decapitated and defoliated shoot tips) were used in the experiment, which were placed on the media vertically, while defoliated shoot tips were placed horizontally or vertically upside down. The explants were cultivated on a Murashige and Skoog (MS) medium supplemented with 1 mg·dm–3 benzyladenine (BA) and 0.5 mg·dm–3 indole-3-butyric acid (IBA). It was noted that both explants orientation and positioning, influenced the multiplication rate. Defoliated shoot tips placed horizontally were characterized by higher multiplication rate (6.8) in comparison to upside down vertical positioning (3.2). It was also observed that removal of shoot apex improved axillary branching, while defoliation of shoots placed in a normal position reduced multiplication rate.

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5

Yadava, U. L., and S. K. Dhir. "In Vitro Regeneration of Trichosanthes from Shoot Tips." HortScience 30, no. 4 (July 1995): 871C—871. http://dx.doi.org/10.21273/hortsci.30.4.871c.

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The morphogenetic potential of parval or pointed gourd (Trichosanthes dioica Roxb.) shoot-tip explants was investigated to establish this species as a model tissue culture system. An effective multiple-shoot propagation method is described. Ten-millimeter shoot tips from young branches of greehouse-grown plants served as explants. They were initiated on a MS basal medium. Multiple shoots were encouraged by transferring established explants to a proliferation medium consisting of MSB + 1 mg BAP/liter, because lower concentrations of BAP (0.1 to 0.5 mg–liter–1) inhibited multiple shoot formation; however, the same concentrations promoted rooting in explants. Medium supplemented with 1 mg BAP/liter and 100 mg PVP/liter caused the best proliferation of shoot tips. Upon transferring to fresh medium of the same composition, these shoot tips elongated 24 cm with three to five nodes in 4 weeks of culturing. Shoot multiplication cultures were maintained by transferring segments of multiple-shoot clusters to medium containing 1 mg BAP/liter and 0.5 mg GA3/liter. Medium supplemented with TDZ inhibited the number of regenerating explants but enhanced the number of shoot buds. Eighty percent of these plantlets were successfully rooted on MS medium supplemented with 1 mg NAA/liter. Plantlets survived in potting soil and exhibited normal growth under mist in the greenhouse.

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6

Towill, Leigh E., and Gayle M. Volk. "(19) Cryopreservation of Arabidopsisthaliana Shoot Tips." HortScience 40, no. 4 (July 2005): 1067E—1067. http://dx.doi.org/10.21273/hortsci.40.4.1067e.

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Arabidopsisthaliana shoot tips provide a model to study processes important for cryopreservation. Cryopreservation was accomplished using both vitrification and two-step cooling methods. With vitrification methods, shoot formation after liquid nitrogen (LN) exposure was as high as 100% and 95% for shoot tips exposed to PVS2 at 0 °C and to PVS3 at 23 °C, respectively. A two-step cooling method also gave greater than 90% survival if shoot tips were cooled at 0.3 °C per minute to below –30 °C before immersing the samples into LN. The high levels of shoot formation after LN exposure in Arabidopsis thaliana shoot tips will allow the use of mutants to examine how alterations in biochemical, metabolic, and developmental processes affect survival and growth.

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7

Pennycooke, Joyce C., and Leigh E. Towill. "Cryopreservation of Sweetpotato Shoot Tips by Vitrification." HortScience 32, no. 3 (June 1997): 472B—472. http://dx.doi.org/10.21273/hortsci.32.3.472b.

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Cryopreservation offers the simplest and most economical way for the long-term conservation of germplasm and vitrification is the preferred method to accomplish this. Undefined endogenous compounds are produced during plant growth and shoot tip preculture conditions. These may influence “cryopreservability” and interact with cryoprotectants that are artificially added during the cryogenic protocol. We are beginning to examine these aspects to improve cryopreservation. Nodal segments of PI 296057 were propagated on a hormone-free modified Murashige and Skoog (MS) solid medium and were grown with 16 hr/8 hr photoperiod. Shoot tips were excised at 0, 3 or 10 hr in light after the dark period. Excised shoot tips were precultured in 0.06 M sucrose in MS for 24 hr and 0.3 M sucrose in MS for 24 hr and then treated with 0.4 M sucrose plus 2 M glycerol for 20 min or 1 hr before being dehydrated in PVS2 [30% (w/v) glycerol, 15% (w/v) ethylene glycol and 15% (w/v) dimethylsulfoxide in MS and 0.4 M sucrose[for 10, 16 or 26 min at 22°C. Shoot tips were placed on thin strips of aluminum foil, which were folded to enclose the shoot tips and then immersed in a liquid nitrogen (LN) slush. Rapid warming and dilution were achieved by transferring the foil strips from LN into 3 ml of 1.2 M sucrose at 22°C for 20 min. All cultures were incubated in darkness for 2 days then dim light for 3 days before transfer to the usual light intensity. Elimination of iron and nitrogen from MS medium in post thaw culture for 5 days increased the viability of LN-treated samples. Maximum survival after LN exposure was achieved with excision immediately after the dark photoperiod, cultured for 1 hr in 0.4 M sucrose plus 2 M glycerol and exposed for 16 min in 100% PVS2 at 22°C. Previously, Towill and Jarret (1992, Plant Cell Reports 11: 175–178) reported that surviving shoot tips developed callus and a variable percentage subsequently formed shoots. In this line all surviving shoot tips eventually formed shoots.

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8

Diettrich, B., P. Donath, A. S. Popov, R. G. Butenko, and M. Luckner. "Cryopreservation of Chamomilla recutita Shoot Tips." Biochemie und Physiologie der Pflanzen 186, no. 1 (January 1990): 63–67. http://dx.doi.org/10.1016/s0015-3796(11)80294-5.

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9

Hitmi, Adnane, Chantal Barthomeuf, and Huguette Sallanon. "Cryopreservation of Chrysanthemum cinerariaefolium Shoot Tips." Journal of Plant Physiology 156, no. 3 (March 2000): 408–12. http://dx.doi.org/10.1016/s0176-1617(00)80081-4.

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10

Hasan, M. F., and B. Sikdar. "In vitro Propagation of Polygonum hydropiper L. from Shoot Tips." Plant Tissue Culture and Biotechnology 20, no. 1 (August 30, 2010): 73–79. http://dx.doi.org/10.3329/ptcb.v20i1.5970.

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An efficient protocol for plant regeneration through multiple shoots induction from shoot tips of Polygonum hydropiper (L.) was established. The highest percentage (96.6) of multiple shoot induction and number of shoots (9.0) per culture were found on MS supplemented with 2.0 mg/l Kn. The induced shoots were excised and inoculated on to MS contains different concentrations of IBA or NAA for rooting. The highest percentage (90.0) of root induction and the highest number of roots per shoot (12.0) was found on MS having 1.0 mg/l IBA. Well rooted plantlets were acclimated properly and transplanted in the soil under natural condition, where cent per cent plantlets survived and grew successfully. Key words: Polygonum hydropiper, Shoot tips, In vitro propagation D.O.I. 10.3329/ptcb.v20i1.5970 Plant Tissue Cult. & Biotech. 20(1): 73-79, 2010 (June)

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11

Brand, Mark H. "THE EFFICACY OF BENZYLADENINE IN OVERCOMING ENDOGENOUS FLUCTUATIONS IN SHOOT TIP CULTURE INITIATION." HortScience 27, no. 6 (June 1992): 697b—697. http://dx.doi.org/10.21273/hortsci.27.6.697b.

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Although it is commonly recommended that shoot tip cultures be initiated from actively growing shoots, it has been demonstrated that shoot tips collected during the period of rapid shoot extension fail to produce shoot proliferating cultures. Shoot tips of Halesia Carolina and Malus `Golden Delicious' were collected at 2 week intervals from budbreak to summer dormancy and placed on medium containing 0, 4.5, 11.0, 22.5 and 44.5 uM benzyladenine (BA) to determine if elevated BA concentrations could overcome seasonal patterns of shoot proliferation potential (SPP). Both species reached maximum SPP 4 weeks post-budbreak (PBB), and exhibited a second window of high SPP during weeks 10 and 12. Elevated BA concentrations failed to overcome poor SPP exhibited by shoot tips harvested 6 to 8 weeks PBB. Shoot tips collected at 10 to 12 weeks PBB responded more favorably to higher exogenous BA concentrations than shoot tips collected at 2, 4, 6, or 8 weeks PBB. It appears as though seasonal fluctuations in SPP involve other endogenous factors in addition to cytokinins.

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12

Touchell, DH, KW Dixon, and B. Tan. "Cryopreservation of Shoot-Tips of Grevillea scapigera (Proteaceae): a Rare and Endangered Plant From Western Australia." Australian Journal of Botany 40, no. 3 (1992): 305. http://dx.doi.org/10.1071/bt9920305.

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Cryopreservation of leaf, petiole, stem pieces and shoot-tips was investigated as a means for long-term maintenance of germplasm of the rare and endangered species, Grevillea scapigera A.S. George. Crypreservation was only achieved using shoot-tips or axillary buds and a slow-cooling regime with the aid of an improvised freezing device. The highest survival of thawed explants (20%) was obtained with 4-week-old in vitro shoot-tips cultured for 48 h in a prefreezing liquid culture medium supplemented with 5% dimethylsulfoxide. The pretreated shoot-tips were frozen in a liquid medium containing 10% dimethylsulfoxide cooled at a rate of 0.5°C/min to -40°C and held at this temperature for 30 min before being plunged into liquid nitrogen. Shoot-tips that survived the freeze-thaw cycle produced callus followed by shoot production 22 weeks after thawing. All shoots regenerated from thawed tissues and transferred to soil appeared phenotypically identical to untreated control shoots and plants. Rapid methods for assessing post-thaw tissue viability and explant recovery using triphenyltetrazolium chloride were tried but these methods were inadequate for determining the capacity of thawed tissues to recover from freeze damage.

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13

Zhao, Lei, Min-Rui Wang, Zhen-Hua Cui, Long Chen, Gayle M. Volk, and Qiao-Chun Wang. "Combining Thermotherapy with Cryotherapy for Efficient Eradication of Apple stem grooving virus from Infected In-vitro-cultured Apple Shoots." Plant Disease 102, no. 8 (August 2018): 1574–80. http://dx.doi.org/10.1094/pdis-11-17-1753-re.

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Apple stem grooving virus (ASGV), a difficult-to-eradicate virus from apple propagative materials, causes serious damage to apple production. The use of virus-free plants has been and is an effective strategy for control of plant viral diseases. This study aimed to eradicate ASGV from virus-infected in-vitro-cultured shoots of four apple cultivars and one rootstock by combining thermotherapy with cryotherapy. In vitro stock shoots infected with ASGV were thermo-treated using an alternating temperature of 36°C (day) and 32°C (night). Shoot tips were excised from the treated stock shoots and subjected to cryotherapy. Results showed that, although thermotherapy did not influence shoot survival rates, it reduced shoot growth and proliferation of in vitro shoots. Shoot regrowth rates decreased while virus eradication frequencies increased in cryo-treated shoot tips as time durations of thermotherapy increased from 0 to 6 weeks. Shoot regrowth and frequency of virus eradication were positively and negatively correlated, respectively, with the size of shoot tips. The protocol established here yielded shoot regrowth rates and virus eradication frequencies of 33 to 76% and 30 to 100%, respectively, in the four apple cultivars and one rootstock. Thermotherapy altered virus distribution patterns, subsequently resulting in production of a larger virus-free area in the thermo-treated shoot tips. Many cells in the top layers of apical dome and some cells in the youngest leaf primordia survived in cryo-treated shoot tips; these cells were most likely free of virus infection. Thus, plants regenerated from the procedure of combining thermotherapy with cryotherapy were free of ASGV, as judged by reverse-transcription polymerase chain reaction. To the best of our knowledge, this is the widest-spectrum technique reported thus far for the production of ASGV-free plants and provides a novel biotechnology for the production of virus-free plants in Malus spp.

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14

Pence, Valerie C. "Cryopreservation of Shoot Tips of Selaginella uncinata." American Fern Journal 91, no. 1 (January 2001): 37–40. http://dx.doi.org/10.1640/0002-8444(2001)091[0037:costos]2.0.co;2.

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15

Towill, Leigh E. "CRYOPRESERVATION OF PAPAYA SHOOT TIPS BY VITRIFICATION." HortScience 25, no. 9 (September 1990): 1066b—1066. http://dx.doi.org/10.21273/hortsci.25.9.1066b.

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Papaya shoot tips, obtained either from seedlings or from in vitro plants, survived liquid nitrogen (-196°C) exposure using a vitrification procedure. Vitrification is a technically simple method but requires large concentrations of cryoprotectants. These were added in two steps, first slow addition of dimethylsulfoxide (DMSO) and PEG-8000, and subsequent fast addition of ethylene glycol (PG). The final concentration before cooling was 40% EG, 7.8% DMSO, and 10% PEG-8000. Both rapid cooling and rapid warming rates were required. Differential scanning calorimetry (DSC) was used to determine that the external solution vitrified upon cooling. It could not be demonstrated by DSC that cells within the shoot-tip vitrified, but since both DMSO and EG rapidly permeate plant cells, vitrification within the cells seems a likely explanation for retention of viability.

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16

Chartier-Hollis, J. M., W. Harris, and P. T. Lynch. "CRYOPRESERVATION OF SHOOT TIPS OF ROSA MULTIFLORA." Acta Horticulturae, no. 424 (July 1996): 367–68. http://dx.doi.org/10.17660/actahortic.1996.424.70.

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17

Li, Jing-Wei, Munetaka Hosokawa, Tomoyuki Nabeshima, Ko Motoki, Haruka Yamada, and Qiao-Chun Wang. "Cryopreservation of viroid-infected chrysanthemum shoot tips." Scientia Horticulturae 244 (January 2019): 1–9. http://dx.doi.org/10.1016/j.scienta.2018.09.004.

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18

Channuntapipat, Chockpisit, Graham Collins, Terry Bertozzi, and Margaret Sedgley. "Cryopreservation ofin vitroalmond shoot tips by vitrification." Journal of Horticultural Science and Biotechnology 75, no. 2 (January 2000): 228–32. http://dx.doi.org/10.1080/14620316.2000.11511228.

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19

Fukai, Seiichi, Masanori Goi, and Michio Tanaka. "Cryopreservation of shoot tips of Caryophyllaceae ornamentals." Euphytica 56, no. 2 (July 1991): 149–53. http://dx.doi.org/10.1007/bf00042058.

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20

Bouafia, S., N. Jelti, G. Lairy, A. Blanc, E. Bonnel, and J. Dereuddre. "Cryopreservation of potato shoot tips by encapsulationdehydration." Potato Research 39, no. 1 (March 1996): 69–78. http://dx.doi.org/10.1007/bf02358208.

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21

Dang, Giang Thi Quynh, Bac Thien Van, and Hoang Ngo Phan. "CULTURING THE SHOOT TIP FROM TUBER SHOOTS OF CALLA LILY (Zantedeschia eliottiana Engl.)." Science and Technology Development Journal 12, no. 17 (November 15, 2009): 64–70. http://dx.doi.org/10.32508/stdj.v12i17.2368.

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Tuber shoots are excised and cultured onto MS medium (Murashige & Skoog, 1962). Shoot tips are isolated from in vitro plantlets and grow in a stable way on medium supplemented with different plant growth regulations, among the medium, shoot tips arise most on MS medium supplemented with 2mg/l BA and 0.5mg/l IBA. Role of endogenous hormones, respiration rate and origin of shoot formation were analysis.

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22

Yamamoto, Shin-ichi, Kuniaki Fukui, Tariq Rafique, Nayyar Iqbal Khan, Carlos Roman Castillo Martinez, Kentaro Sekizawa, Toshikazu Matsumoto, and Takao Niino. "Cryopreservation of in vitro-grown shoot tips of strawberry by the vitrification method using aluminium cryo-plates." Plant Genetic Resources 10, no. 1 (November 28, 2011): 14–19. http://dx.doi.org/10.1017/s1479262111000906.

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Cryopreservation using an aluminium cryo-plate was successfully applied to in vitro-grown strawberry (Fragaria × ananassa Duch.) shoot tips. The shoots were cold-hardened at 5°C for 3 weeks with an 8-h photoperiod. The shoot tips (1.5–2.0 mm × 0.5–1.0 mm) were dissected from the shoot and pre-cultured at 5°C for 2 d on Murashige and Skoog medium containing 2 M glycerol and 0.3 M sucrose. The pre-cultured shoot tips were placed on the aluminium cryo-plate containing ten wells embedded in alginate gel. Osmoprotection was performed by immersing the cryo-plates in a loading solution (2 M glycerol and 0.8 M sucrose) for 30 min at 25°C. Dehydration was performed by immersing the cryo-plates in plant vitrification solution 2 for 50 min at 25°C. Then, the cryo-plate with shoot tips was transferred into an uncapped cryotube that was held on a cryo-cane and directly immersed into liquid nitrogen (LN). After storage in LN, shoot tips attached to the cryo-plate were directly immersed into 2 ml of a 1 M sucrose solution for regeneration. Using this procedure, the average regrowth level of vitrified shoot tips of 15 strawberry cultivars reached 81%. This new method has many advantages and will facilitate the cryostorage of strawberry germplasm.

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23

Chang, Yongjian, and Barbara M. Reed. "Preculture Conditions Influence Cold Hardiness and Regrowth of Pyrus cordata Shoot Tips After Cryopreservation." HortScience 36, no. 7 (December 2001): 1329–33. http://dx.doi.org/10.21273/hortsci.36.7.1329.

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Cold hardiness and cryogenic survival of micropropagated pear (Pyrus cordata Desv.) shoots were evaluated after pretreatments with ABA and sucrose. Shoot cold hardiness increased by 3 °C, and cryopreserved shoot tip growth increased by 17% after a 4-week 150 μm ABA pretreatment. Low temperature (LT) pretreatments improved the recovery of cryopreserved P. cordata shoot tips. Six to 10 weeks of LT were required for reaching high cryopreservation recovery. ABA and LT treatments produced significant synergistic effects on both cold hardiness and cryopreservation recovery. ABA shortened the LT requirement for high cryopreservation growth from 10 to 2 weeks. The optimal treatment for recovery of cryopreserved shoot tips was a 3 week culture on 50 μm ABA followed by 2 weeks of LT, while the maximum cold hardiness (-22.5 °C) was obtained with 150 μm ABA and 2-week LT. A 4 week culture on 150 μm ABA at 25 °C induced dormancy in 74% of shoot tips, but had little effect on cryopreservation growth unless combined with LT. Control and ABA-treated shoot tips, lateral buds, and leaves had similar cold hardiness (-10 to -12 °C), but LT and LT+ABA-treated shoot tips survived the lowest temperatures (-17 to -23 °C), lateral buds next (-15 to -20 °C), and finally leaves (-14 to -18 °C). An increase in the preculture-medium sucrose concentration from 2% to 7% combined with 2-week LT significantly increased cryopreserved shoot tip growth (0% to 75%) and decreased the LT50 from -7.8 to -12.4 °C. The optimal shoot pretreatment for successful recovery of cryopreserved P. cordata shoot tips was a 3 week culture on either 50 μm ABA or 5% to 7% sucrose medium followed by 2 weeks of LT, and increased shoot tip growth from zero to >70%. Chemical name used: abscisic acid (ABA).

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24

Piątczak, Ewelina, and Halina Wysokińska. "In vitro regeneration of Centaurium erythraea Rafn from shoot tips and other seedling explants." Acta Societatis Botanicorum Poloniae 72, no. 4 (2011): 283–88. http://dx.doi.org/10.5586/asbp.2003.036.

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Various explants from 30-day-old seedlings of <em>Centaurium erythraea </em>Rafn were evaluated for their morphogenetic capacity under in vitro culture conditions. Shoot formation from shoot tip explants was achieved mainly through adventitious bud differentiation. The highest number of shoots (up to 43.3 ± 2.2 from a single shoot tip) was obtained on Murashige and Skoog medium (MS) supplemented with indole-3-acetic acid (IAA) (0.57 μM) and 6-benzylaminopurine (BAP) (4.4 μM). Adventitious shoot regeneration was also achieved through organogenesis from calluses obtained from hypocotyls, cotyledons, roots and leaves on MS medium containing IAA (2.85 μM) and BAP (0.88 μM). Significant differences were noted between explant types in their effects on shoot regeneration. In the primary culture, the best response was obtained either from calluses derived from roots or leaves (44.4 ± 4.5 and 40.2 ± 6.0 shoots per callus, respectively). The number of subcultures of inoculated calluses affected both the multiplication rate (the number of shoots/explant) and shoot morphology (the frequency of shoot hyperhydricity). Shoots rooted with the frequency of 94-100% after culture on MS medium without growth regulators. Plantlets were successfully acclimatized (97%) under high relative humidity and then moved to the greenhouse.

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25

Niino, T., and A. Sakai. "CRYOPRESERVATION OF ALGINATE-COATED IN-VITRO-GROWN SHOOT TIPS OF APPLE." HortScience 27, no. 6 (June 1992): 695e—695. http://dx.doi.org/10.21273/hortsci.27.6.695e.

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In-vitro-grown shoot tips of apple (Malus domestia Borkh cv. Fuji) were successfully cryopreserved by dehydration of alginate-coated shoot tip. Cold-hardened shoot tips (at 5°C for 3 weeks) were precultured on a medium containing increasing concentrations of sucrose. The shoot tips were trapped into alginate coated beads containing 0.5M sucrose followed by preculture in a medium supplemented with 1.0M sucrose. Beads containing 1 shoot tip were dehydrated up to about 32% on sterile dry silica gel at 25°C followed by a plunge in LN. After rapid warming, approximately 80% shoot formation was achieved. This encapsulation-dehydration technique may permit strage of shoot tips at higher temperatures than that of LN.

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26

Nogueira, Gabriela Ferreira, Moacir Pasqual, and Jonny Everson Scherwinski-Pereira. "Survival of sugarcane shoot tips after cryopreservation by droplet-vitrification." Pesquisa Agropecuária Brasileira 48, no. 11 (November 2013): 1524–27. http://dx.doi.org/10.1590/s0100-204x2013001100014.

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The objective of this work was to evaluate the phytotoxicity of a plant vitrification solution (PVS2), and the survival of shoot tips of the sugarcane variety SP716949, after cryopreservation by droplet-vitrification. Shoot tips were precultured for 24 hours in MS medium containing 0.3 mol L-1 sucrose, and exposed to PVS2 for 0, 20 or 30 min. Shoot tips were then immersed in liquid nitrogen. Thawing was fast in concentrated sucrose solution (1.2 mol L-1). PVS2 is a nontoxic to shoot tips, which in turn are sensitive to liquid nitrogen. The best results occurred when shoot tips were maintained for up to 20 min in PVS2 solution, before freezing, with 20% survival.

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27

Kausar, M., S. Parvin, ME Haque, M. Khalekuzzaman, B. Sikdar, and MA Islam. "Efficient Direct Organogenesis from Shoot Tips and Nodal Segments of Ash Gourd (Benincasa Hispida L.)." Journal of Life and Earth Science 8 (August 22, 2014): 17–20. http://dx.doi.org/10.3329/jles.v8i0.20135.

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The effect of external application of phytohormone on explants viz., shoot tips and nodal segments of ash gourd (Benincasa hispida L.) was tested on the frequency of shoot and root induction. Shoot tips and nodal segments were cultured on MS medium supplemented with different concentrations and combinations of cytokinins (BAP, Kinetin), auxin (IBA, NAA) and gibberellic acid (GA3) for multiple shoot formation and root induction. The highest number (up to 90%) of multiple shoot formation was obtained from the shoot tips in MS medium fortified with 1.5 mg/l BAP + 0.2 mg/l GA3, where average number of shoots per culture was 5.55. In case of nodal segment, better response (up to 78%) for shoot multiplication was found in MS with 2.0 mg/l BAP + 0.2 mg/l GA3. The concentration of 1.0 mg/l IBA was found to be most effective for root initiation in microshoot developed from both types of explants. Rooted plantlets were acclimatatized and established in sandy soil with good survival rate DOI: http://dx.doi.org/10.3329/jles.v8i0.20135 J. Life Earth Sci., Vol. 8: 17-20, 2013

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28

Tanaka, Daisuke, Takao Niino, Yoshiko Tsuchiya, Kazuto Shirata, and Matsuo Uemura. "Cryopreservation of shoot tips of endangered Hayachine-usuyukiso ( Leontopodium hayachinense (Takeda) Hara et Kitam.) using a vitrification protocol." Plant Genetic Resources: Characterization and Utilization 6, no. 02 (May 15, 2008): 164–66. http://dx.doi.org/10.1017/s1479262108993175.

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Hayachine-usuyukiso (Leontopodium hayachinense) is an alpine plant native to Mt Hayachine. This unique chrysanthemum is listed as an endangered plant by the Department of Conservation, Iwate Prefecture, and as a threatened plant by the Ministry of the Environment, Japan. We successfully cryopreserved the shoot tips fromin vitro-grownL. hayachinenseshoots using a vitrification protocol. Cold-hardened shoot tips were excised and pre-cultured on a solidified Murashige–Skoog medium containing 0.3 M sucrose for 1 d at 5°C. The shoot tips were then treated with loading solution for 20 min at 25°C, dehydrated in plant vitrification solution 2 for 120 min at 25°C and immersed in liquid nitrogen. The survival rate of the vitrified shoot tips was 63.3% after 30 d of regrowth. This protocol appears to be a promising technique for the cryopreservation ofin vitro-grown shoots of this endangered plant.

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Marković, Zvjezdana, Philippe Chatelet, Isabelle Sylvestre, Jasminka Kontić, and Florent Engelmann. "Cryopreservation of grapevine (Vitis vinifera L.) in vitro shoot tips." Open Life Sciences 8, no. 10 (October 1, 2013): 993–1000. http://dx.doi.org/10.2478/s11535-013-0223-8.

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AbstractIn this work, we compared the efficiency of encapsulation-dehydration and droplet-vitrification techniques for cryopreserving grapevine (Vitis vinifera L.) cv. Portan shoot tips. Recovery of cryopreserved samples was achieved with both techniques; however, droplet-vitrification, which was used for the first time with grapevine shoot tips, produced higher regrowth. With encapsulationdehydration, encapsulated shoot tips were precultured in liquid medium with progressively increasing sucrose concentrations over a 2-day period (12 h in medium with 0.25, 0.5, 0.75 and 1.0 M sucrose), then dehydrated to 22.28% moisture content (fresh weight). After liquid nitrogen exposure 37.1% regrowth was achieved using 1 mm-long shoot tips and only 16.0% with 2 mm-long shoot tips. With droplet-vitrification, 50% regrowth was obtained following treatment of shoot tips with a loading solution containing 2 M glycerol + 0.4 M sucrose for 20 min, dehydration with half-strength PVS2 vitrification solution (30% (w/v) glycerol, 15% (w/v) ethylene glycol, 15% dimethylsulfoxide and 0.4 M sucrose in basal medium) at room temperature, then with full strength PVS2 solution at 0°C for 50 min before direct immersion in liquid nitrogen. No regrowth was achieved after cryopreservation when shoot tips were dehydrated with PVS3 vitrification solution (50% (w/v) glycerol and 50% (w/v) sucrose in basal medium).

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Tetsumura, Takuya, and Hisajiro Yukinaga. "Comparative Rooting of Shoot Tips of Four Japanese Persimmon Cultivars vs. Shoots Regenerated from Roots Cultured In Vitro." HortScience 35, no. 5 (August 2000): 940–44. http://dx.doi.org/10.21273/hortsci.35.5.940.

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When cultured in vitro, roots of four Japanese persimmon (Diospyros kaki L.) cultivars formed adventitious shoots on MS medium with 10 μm zeatin and 0.01 μm indole-3-acetic acid, although their organogenetic capacities varied. Histological study revealed that the origin of the adventitious shoots was the pericycle. The regenerated shoots grew well on the shoot proliferation medium (MS with 5 μm zeatin). Final rooting percentages of shoots regenerated from roots of three of the four cultivars were greater than those of shoots that originated from shoot tips and that had been subcultured >50 times. Shoots regenerated from `Jiro' roots rooted 10 days earlier, had more roots than those from shoot tips, and maintained higher rooting ability over ten subcultures. Rooted `Hiratanenashi' shoots regenerated from roots survived better after acclimatization than those from shoot tips. No obvious variants were observed either in vitro or in the field. The trees regenerated from roots flowered within 4 years. These findings suggest that partial rather than true rejuvenation was responsible for both the early flowering and the juvenile characteristics, i.e., the enhanced rooting ability, observed in the regenerated plants. Chemical name used: 6-(4-hydroxy-3-methylbut-2-enylamino) purine (zeatin).

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Zou, Li-Juan, Jin-Yao Hu, Ming-Hua Luo, and Qing-Gui Wu. "In vitro propagation of the Chinese traditional and medicinal plant Heracleum scabridum Franch." Bangladesh Journal of Botany 48, no. 3 (September 30, 2019): 575–81. http://dx.doi.org/10.3329/bjb.v48i3.47933.

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Three explants such as stem tips, leaves and petioles of Heracleum scabridum were compared for their shoot development/differentiation ability by using different plant growth regulators (PGRs). The most effective PGRs combination for callus induction and organogenesis was determined. TDZ, Kn, Zn and IAA were used to induce multiple shoots. For indirect organogenesis (from the calli), the best response (27.25 ± 4.19 shoot per stem tips) and (18.23 ± 2.12 per leaves) were obtained in Murashige and Skoog (MS) medium fortified with 0.5 mg/l IAA and 3.0 mg/l Zn with 100, 97.3% induction rate, respectively. MS medium containing 0.5 mg/l IAA and 3.0 mg/l Zn was also found to be optimal for shoot regeneration from petioles. The highest percentage of regeneration (94.6) with mean number of shoots 17.83 ± 4.87 from petioles were produced. For direct organogenesis (from axillary bud shoot clumps), 0.1 mg/l IAA and 1.5 mg/l TDZ were found to be optimal for shoot regeneration of stem tips, with mean numbers of axillary bud shoot clumps 7.12 ± 1.23 were produced. Plantlets were transferred to soil with 95% of plants acclimatized recovered successfully.

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Pan, C. P., Y. Q. Wang, J. Liu, Q. X. Deng, and L. Tao. "PLANT REGENERATION FROM SHOOT TIPS IN TRIPLOID LOQUAT." Acta Horticulturae, no. 1092 (August 2015): 97–103. http://dx.doi.org/10.17660/actahortic.2015.1092.15.

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FUKAI, Seiichi. "Freeze preservation of dianthus shoot tips by programfreezer." Environment Control in Biology 25, no. 2 (1987): 25–30. http://dx.doi.org/10.2525/ecb1963.25.25.

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Jevremović, S., A. Subotić, C. Benelli, A. De Carlo, and M. Lambardi. "CRYOPRESERVATION OF IRIS PUMILA SHOOT TIPS BY VITRIFICATION." Acta Horticulturae, no. 908 (September 2011): 355–59. http://dx.doi.org/10.17660/actahortic.2011.908.47.

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35

Damiano, C., A. Sgueglia, M. Arias, A. Frattarelli, E. Condello, and E. Caboni. "CRYOPRESERVATION OF PEACH SHOOT TIPS BY ENCAPSULATION DEHYDRATION." Acta Horticulturae, no. 918 (December 2011): 121–24. http://dx.doi.org/10.17660/actahortic.2011.918.13.

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Wang, Qiaochun, and Jari P. T. Valkonen. "Cryotherapy of shoot tips: novel pathogen eradication method." Trends in Plant Science 14, no. 3 (March 2009): 119–22. http://dx.doi.org/10.1016/j.tplants.2008.11.010.

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Monette, Paul L. "Micropropagation of kiwifruit using non-axenic shoot tips." Plant Cell, Tissue and Organ Culture 6, no. 1 (1986): 73–82. http://dx.doi.org/10.1007/bf00037760.

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Lynch, P. T., W. C. Harris, and J. M. Chartier-Hollis. "The cryopreservation of shoot tips of Rosa multiflora." Plant Growth Regulation 20, no. 1 (October 1996): 43–45. http://dx.doi.org/10.1007/bf00024056.

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Smyda-Dajmund, Paulina. "Cryopreservation of Shoot Tips and Pollen of Potato." Plant Breeding and Seed Science 76, no. 1 (December 20, 2017): 75–80. http://dx.doi.org/10.1515/plass-2017-0025.

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Abstract Cryopreservation is a frequently used method of long-term storage of potato meristems and pollen in liquid nitrogen (LN) in temperature of -196°C. This technique allows for theoretically unlimited storage of potato material. The most popular method of potato shoot tips preservation is cryopreservation by the solidification of liquids without crystallization (vitrification).The best method of pollen conservation is its direct immersion in LN. The successful regeneration after vitrification is genotype-dependent, which require optimization of protocol.

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Teixeira, Aline S., M. Elena González-Benito, and Antonio D. Molina-García. "Glassy State and Cryopreservation of Mint Shoot Tips." Biotechnology Progress 29, no. 3 (March 27, 2013): 707–17. http://dx.doi.org/10.1002/btpr.1711.

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Yu, Rongpei, Ying Cheng, Yanfei Pu, Fan Li, and Shugang Lu. "In Vitro Propagation of Resurrection Plant Selaginella pulvinata Using Frond Tips as Explants." HortScience 56, no. 3 (March 2021): 313–17. http://dx.doi.org/10.21273/hortsci15546-20.

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The resurrection plant Selaginella pulvinata (Hook. & Grev.) Maxim is used as an ornamental and medicinal plant. It is also a good candidate for exploring the desiccation tolerance of resurrection plants. However, there is not an efficient propagation method for S. pulvinata. In the present study, we evaluated the establishment of in vitro propagation of S. pulvinata using frond tips as explants. The original shoot induction, adventitious shoot proliferation and plantlet growth media, and substrate type of plantlet acclimatization were investigated. The highest induction rate of original shoots (61.77 ± 5.17%) was obtained on half-strength (1/2) MS medium supplemented with 0.1 mg·L−1 N6-benzylaminopurine (BAP). The 1/2 MS with 1.0 mg·L−1 BAP was the most effective medium for the adventitious shoot proliferation. The quarter-strength (1/4) MS containing 0.1% (w/v) active charcoal (AC) was optimum for plantlets proliferated from adventitious shoots and plantlet growth. Approximately 98 plantlets could be obtained from one single original shoot via one-time shoot proliferation cultivation and plantlet cultivation. The acclimated plants on a 5:1 (v/v) mixture of peat and perlite had the highest survival rate (92.13 ± 1.67%). The acclimated plants maintained excellent resurrection ability.

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Rehana, S., MS Alam, KS Islam, and MA Samad. "Influence of Growth Regulators on Shoot Proliferation and Plantlet Production from Shoot Tips of Banana." Progressive Agriculture 20, no. 1-2 (November 4, 2013): 9–16. http://dx.doi.org/10.3329/pa.v20i1-2.16840.

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The effect of BAP and IBA on in vitro regeneration of four banana cultivars viz. Amritsagar, Seeded banana, Sabri and Anajee was studied. The response of single shoot regeneration from shoot tips of four banana cultivars at different concentrations of BAP was found to be different. The cultivar Amritsagar produced the highest percentage (60 %) of single shoot at 4.0 mg/l BAP within 10-15 days. The cultivar Sabri and Anajee produced lower percentage of single shoots. Rates of shoot multiplication of Amritsagar, Sabri, and Anajee were 6-7 plantlets/explant, 2-4 plantlets/explant, and 2 plantlets/explant, respectively on medium containing 4.0 mg/l BAP after 30 days of culture. But in subsequent subculture, on the same medium, Amritsagar produced the highest number of plantlets (9 plantlets/explant) within the same period of time. The best root formation in multiplied shoots of Amritsagar was found on MS medium containing 2.0 mg/l IBA after 15 days of culture. All the in vitro cultured banana plantlets of Amritsagar survived when weaned to ex vitro conditions on soil.DOI: http://dx.doi.org/10.3329/pa.v20i1-2.16840 Progress. Agric. 20(1 & 2): 9 16, 2009

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Li, Bai-Quan, Chao-Hong Feng, Ling-Yun Hu, Min-Rui Wang, Long Chen, and Qiao-Chun Wang. "Shoot regeneration and cryopreservation of shoot tips of apple (Malus) by encapsulation–dehydration." In Vitro Cellular & Developmental Biology - Plant 50, no. 3 (May 23, 2014): 357–68. http://dx.doi.org/10.1007/s11627-014-9616-2.

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Makowczyńska, Joanna, and Emilia Andrzejewska-Golec. "Micropropagation of Plantago asiatica L. through culture of shoot-tips." Acta Societatis Botanicorum Poloniae 72, no. 3 (2011): 191–94. http://dx.doi.org/10.5586/asbp.2003.025.

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<p>Shoot-tip multiplication of the medicinal species - <em>Plantago asiatica</em> was carried on MS medium with IAA and BAP or kinetin. Best results in micropropagation were achieved by adding 0.1 mg/dm<sup>3</sup> IAA and 1 mg/dm<sup>3</sup> BAP. After 6 weeks shoots were transferred to MS medium for rooting. The resulting plantlets were transferred after 8 weeks into pots and after a period of adaptation into the ground (field culture).</p><p>The species <em>Plantago asiatica </em>was propagated in vitro by shoot-tip multiplication for the first time.</p>

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KWAŚNIEWSKA, Ewelina, Ewa DZIEDZIC, and Bożena PAWŁOWSKA. "Integration of Cryopreservation and Tissue Culture for Germplasm Conservation and Propagation of Rosa pomifera ‘Karpatia’." Notulae Botanicae Horti Agrobotanici Cluj-Napoca 45, no. 1 (June 10, 2017): 208–14. http://dx.doi.org/10.15835/nbha45110566.

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Cryopreservation is an useful technique for long-term conservation that requires minimal space and maintenance. Germplasm protection of Rosa is important to preserve genetic diversity, to store material for breeding and to expand new research. This study was conducted to develop a droplet vitrification cryopreservation and micropropagation of Rosa pomifera cv. ‘Karpatia’, whose large hypanthia are characterized by remarkable pro-health properties. Culture in vitro was stabilized and shoot tips collected from dormant buds served as initial explants. The multiplication of shoots was carried out on MS medium containing benzyladenine. For the droplet vitrification cryopreservation, shoot tips from in vitro cultures were used: small with exposed meristem, and large with a meristem covered with leaves, as well as shoot tips from in situ plants, which were collected in winter. Treatment time with plant vitrification solution (PVS2) was also tested (10-30 minutes). From in vitro culture, 32-41% small explants with exposed meristem survived, but they regenerated at a very low level. The best cryostorage results were obtained for shoot tips from dormant buds and a 20-minute PVS2 treatment: the survival was 84% and regeneration 72%. During the post-freezing regeneration multiplication index was 2.4 shoots per one multiplication cycle, after cryopreservation and in the control. On half MS medium without growth regulators, 97-99% of shoots rooted, and all rooted plants have adapted to ex vitro conditions and were planted into the soil. Biometric analyses during shoot multiplication, rooting and acclimatization stages did not reveal any changes compared to the non-cryopreserved samples.

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Phiman, Nancy, and Michael E. Kane. "876 PB 238 MICROPROPAGATION OF UNIOLA PANICULATA L. (SEA OATS) FROM TILLER EXPLANTS." HortScience 29, no. 5 (May 1994): 559d—559. http://dx.doi.org/10.21273/hortsci.29.5.559d.

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Beach stabilization by replanting dune species such as Uniola paniculata L. (Sea Oats), is an accepted practice to control erosion in the southeastern United States. Increased restrictions on collection of sea oat seed and plant material for propagation is of increasing concern. Development of micropropagation protocols for establishment and production of sea oats from donor plants of known phenotype would be useful for selecting and producing plants with commercially valuable characteristics. Terminal and lateral shoot tips (3 mm wide and 4 mm high) from containerized plants were surface sterilized and established on Linsmaier & Skoog mineral salts and organics supplemented with 87.6 mM sucrose, 2.2 μM benzyladenine solidified with 0.8% TC® Agar. Terminal tiller shoot tips were more responsive than lateral shoot tips. Four monthly subcultures were. required for stabilized shoot multiplication from culture lines established from terminal tiller shoot tips. Shoot organogenesis frequently occurred from the cut leaf surfaces of subcultured shoot clusters. Microcuttings were established ex vitro in plug cells containing sand or vermiculite.

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47

Ružić, Djurdjina, Tatjana Vujović, and Radosav Cerović. "Cryopreservation of cherry rootstock Gisela 5 (Prunus cerasus × Prunus canescens) shoot tips by droplet-vitrification technique." Journal of Horticultural Research 21, no. 2 (December 1, 2013): 79–85. http://dx.doi.org/10.2478/johr-2013-0025.

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ABSTRACT The droplet-vitrification technique was applied to in vitro shoot tips of cherry rootstock Gisela 5 (Prunus cerasus × Prunus canescens). Explants were precultured in the dark at 23 °C, in liquid MS medium with a progressively increasing sucrose concentration (0.3 M for 15 h, then 0.7 M for 5 h). Loading involved a 30 min incubation of explants in a solution comprising 1.9 M glycerol and 0.5 M sucrose. Explants were dehydrated at room temperature using a solution PVS A3 [Murashige and Skoog (MS) liquid medium, 22.5% (w/v) sucrose, 37.5% (w/v) glycerol, 15% (w/v) ethylene glycol and 15% (w/v) dimethylsulfoxide] for 30, 40 and 50 min and the PVS3 solution [MS liquid medium, 50% (w/v) sucrose, 50% (w/v) glycerol] for 60, 90 and 120 min. Explants were cooled by direct immersion in liquid nitrogen (LN) in 10 μl droplets of vitrification solution placed on aluminum foil strips. The foil strips were retrieved from LN and immersed in preheated (37 °C) unloading solution (0.8 M sucrose) for 30 s, and an equal volume of unloading solution at room temperature was added for further incubation for 30 min. Shoot tips were transferred onto the regrowth medium, cultivated in the dark for 7 days before being incubated under standard conditions. Three weeks after transferring the shoot tips onto the regrowth medium, the survival rate of control and cryopreserved explants of Gisela 5 dehydrated with PVS A3 was 100%, regardless of the treatment duration. After dehydration with solution PVS3, the survival varied between 70 and 100% for control explants and 78 and 95% for cryopreserved shoot tips. Gisela 5 shoot tips dehydrated for 40 min with PVS A3 vitrification solution demonstrated the best regrowth (38%). When using the PVS3 solution, survival of cryopreserved shoot tips was the highest (95%) after 60 min treatment followed by 40% regrowth. After three successive subcultures on shoot multiplication, medium shoots recovered viability, multiplication ability and morphology equal of that prior to cryopreservation.

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48

Towill, L. E. "The Effect of Glass-cracking during Cooling in Liquid Nitrogen on Viability of Mint Shoot Tips." HortScience 30, no. 4 (July 1995): 874D—874. http://dx.doi.org/10.21273/hortsci.30.4.874d.

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Cryopreservation using vitrification has been reported for several plant species. Shoot tips and vitrification solution were placed in semen straws and immersed in liquid nitrogen (LN). Cracking of the external glass occurred, but may be avoided by annealing slightly below the glass transition temperature before immersion. A varying percentage still cracked with some vitrification solutions. Rapid warming also can cause cracking. There is concern that cracking may reduce viability. Shoot tips from Mentha species were used to examine this problem. Glass cracking during either cooling or warming did not produce visible damage to shoot tips. Viability of shoot tips from tubes that cracked during cooling was not different from those that did not crack; however, shoot formation was slightly reduced. Cracking upon warming did not reduce viability nor shoot formation. Very slow warming reduced viability, but warming in either water or air (room temperature) gave higher levels of survival.

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49

Uchendu, Esther, Hemant Lata, Suman Chandra, Ikhlas A. Khan, and Mahmoud A. ElSohly. "Cryopreservation of Shoot Tips of Elite Cultivars of Cannabis sativa L. by Droplet Vitrification." Medical Cannabis and Cannabinoids 2, no. 1 (April 4, 2019): 29–34. http://dx.doi.org/10.1159/000496869.

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Cannabis sativa L. (marijuana or hemp) is recognized worldwide for its psychoactive properties as well as for fiber production. This study focused on the evaluation of 3 droplet vitrification protocols for long-term conservation of shoot tips in liquid nitrogen (LN). Shoot tips (∼0.5 mm) were excised from 3- to 4-week-old in vitro-grown shoots of 3 cultivars (MX, VI-20, and B-5: high tetrahydrocannabinol [THC], high cannabidiol [CBD], and intermediate THC∼CBD, respectively) and pretreated on 5% dimethyl sulfoxide agar plates for 48 h. The shoot tips were then vitrified in LN using 3 separate cryoprotectant (plant vitrification solutions [PVS] #2, #3, and #4) droplets on an aluminum cryoplate. There was no significant difference between the regrowth of cryopreserved shoot tips exposed to PVS2 for 15 and 20 min, but regrowth of all 3 cultivars significantly declined after 20 min of exposure. Exposure duration of 15 min was adapted for subsequent experiments. Regrowth of cryopreserved MX was significantly higher with PVS2 (63%) than with PVS3 and PVS4 (≤5%). Regrowth of cryopreserved VI-20 was highest with PVS2 (57%) and significantly higher than with PVS3 and PVS4 (≤25%). The regrowth of cryopreserved shoot tips of B-5 was significantly different between all 3 protocols with PVS2 > PVS4 > PVS3. Both PVS2 and PVS4 produced regrowth above 55%, while regrowth with PVS3 was significantly lower (31%). These results indicate that 15–20 min of exposure to PVS2 are most suitable for cryopreservation of these varieties. This is the first report on protocol development for the cryopreservation of organized tissues of C. sativa L. for germplasm conservation.

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Rogers, Suzanne M. D., Kalyani Dias, and David Byrne. "REGENERATION OF CITRUS VIA SHOOT APECIES." HortScience 25, no. 9 (September 1990): 1112a—1112. http://dx.doi.org/10.21273/hortsci.25.9.1112a.

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Viral damage is a major problem in citrus. As most citrus are asexually propagated, it is necessary to have an alternative way of regenerating virus-free plants from infected plants. Shoot apicies are the most suitable explant material for this purpose because that part of the plant is virus-free. Fifty sour orange shoot tips and 22 Swingle shoot tips, 1 mm - 1.5 mm long, were excised from in vitro germinated seedlings and cultured on semisolid Murashige and Skoog medium, without growth regulators, containing 0.2 % Gelrite. After 8-10 weeks, shoots and leaves developed in 68'% of the sour orange explants, and in 77% of the Swingle explants. Some explants produced roots, after 11-12 weeks, and could be removed from culture and established in soil medium.

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