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Journal articles on the topic 'Shoot regeneration'
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1
Tezuka, Takahiro, Masashi Harada, Masahumi Johkan, Satoshi Yamasaki, Hideyuki Tanaka, and Masayuki Oda. "Effects of Auxin and Cytokinin on In Vivo Adventitious Shoot Regeneration from Decapitated Tomato Plants." HortScience 46, no. 12 (December 2011): 1661–65. http://dx.doi.org/10.21273/hortsci.46.12.1661.
Full textAbstract:
Adventitious shoots can be regenerated from the cut surface of the primary shoot and lateral branches in decapitated plants in vivo. This inherent regenerative ability of plants is useful for mass propagation. In the present study, we conducted histological observations of shoot regeneration and applied auxin and cytokinin to decapitated seedlings in four tomato cultivars. The cultivars produced different numbers of adventitious shoots after decapitation; ‘Petit’ produced the largest number of adventitious shoots (78.5 ± 10.2) and ‘Momotaro’ produced the fewest (12.1 ± 3.3). Histological observation of ‘Petit’ revealed that adventitious shoots regenerated from calli formed at the cut surface of stems. Adventitious shoot formation was inhibited by the presence of lateral branches. Shoot regeneration was prevented by application of 1-naphthaleneacetic acid to ‘Petit’. Application of 6-benzyladenine promoted shoot regeneration in ‘Momotaro’. These results suggest auxin synthesized de novo from the lateral branches inhibited shoot regeneration after decapitation and endogenous cytokinin might stimulate shoot regeneration. Chemical names: 1-naphthaleneacetic acid (NAA); 6-benzyladenine (BA)
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Cao, X., and F. Hammerschlag. "307 Growth Regulator Pretreatments Significantly Enhance the Efficiency of Shoot Organogenesis from Leaf Explants of Highbush Blueberry Cultivar Bluecrop." HortScience 35, no. 3 (June 2000): 445A—445. http://dx.doi.org/10.21273/hortsci.35.3.445a.
Full textAbstract:
As part of a program to develop transgenic highbush blueberry (Vaccinium corymbosum L.) cultivars, studies were conducted to determine optimum conditions for high-efficiency shoot regeneration from leaf explants of in vitro propagated, commercially important, tissue culture-recalcitrant `Bluecrop' shoot cultures. The effects of pretreatments, growth regulators, and age of explant source on shoot organogenesis were investigated. A maximum of 98% shoot regeneration and 10 shoots regenerating per leaf explant occurred when explants of 2-week-old shoot cultures were incubated in the dark (for a total of 14 days) on pretreatment medium #1 containing 2.6 μM NAA and 5 μM TDZ for 4 days, next on pretreatment medium #2 containing 2.6 μM NAA and 7 μM zeatin riboside for 3 days, then on regeneration medium containing 1 μM TDZ for 6 weeks, and last on medium without growth regulators for 10 days. No shoot regeneration occurred if explants were incubated without exposure to pretreatments before incubation on regeneration medium. There were no significant differences in percentage of regeneration or the number of shoots regenerating per explant from leaf explants derived from either 1-, 2-, or 3-week-old shoot cultures. Shoot production per explant on 1 μM TDZ was about three times that on either 0.5 μM TDZ or 20 μM zeatin riboside, and nine times that on 5 μM TDZ.
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Chae, Soo Cheon, Haeng Hoon Kim, and Sang Un Park. "Ethylene Inhibitors Enhance Shoot Organogenesis of Gloxinia (Sinningia speciosa)." Scientific World Journal 2012 (2012): 1–4. http://dx.doi.org/10.1100/2012/859381.
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Shoot organogenesis and plant regeneration inSinningia speciosawere improved using ethylene inhibitors. The leaf explants were cultured on initial shoot regeneration media (MS media with BAP at 2 mg/L + NAA at 0.1 mg/L) supplemented with different concentrations of aminoethoxyvinylglycine (AVG), cobalt chloride (CoCl2), and silver thiosulphate (STS). The addition of AVG, CoCl2, and STS significantly improved the regeneration frequency giving higher shoots per explant and longer shoot length. The highest shoot growth was found when STS at 5 mg/L was incorporated with generation medium, performing highest regeneration frequency with highest number of shoots. This treatment (STS at 5 mg/L) produced 40% more shoots per explant compared to control followed by STS at 10 mg/L with increasing 37% more shoots compared to control. In the cases of AVG and CoCl2the highest shoot number per explant was found at 1 mg/L. Treated with AVG and CoCl2at 1 mg/L increased shoot number by 16 and 12%, respectively, compared to control. Ethylene inhibitors could be used as a possible micropropagation and plant transformation protocol inS. speciosafor plant regenerations.
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Xu, L., G. F. Liu, and M. Z. Bao. "Adventitious Shoot Regeneration from In Vitro Leaves of Formosan Sweetgum (Liquidambar formosana L.)." HortScience 42, no. 3 (June 2007): 721–23. http://dx.doi.org/10.21273/hortsci.42.3.721.
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Plantlets were regenerated from in vitro-grown leaf explants of five genotypes of Liquidambar formosana on WPM basal medium supplemented with different concentrations of TDZ and NAA. With the addition of 0.27 μm NAA, regeneration efficiency was increased by 2- to 4-fold over that with TDZ alone. Lower concentrations of TDZ (0.45–2.27 μm) were beneficial for regenerating shoot clusters. Four genotypes (P2, P6, P9, and P11) showed high regeneration rates (up to 90%), whereas genotype P13 showed a low capability for shoot regeneration on all media tested (<35%). For all five genotypes, the optimum medium for inducing adventitious shoots was WPM supplemented with 1.14 μm TDZ and 0.27 μm NAA, on which regeneration rate ranged from 72.6% to 89.5% and adventitious shoot clusters per regenerating leaf explant ranged from 2.63 to 3.11 in four genotypes (P2, P6, P9, and P11), while for P13, the regeneration rate and number of shoot clusters per regenerating explant were 23% and 1.39, respectively. Transfer of shoot clusters to WPM basal medium containing 0.54 μm NAA, 2.22 μm BA, and 1.44 μm GA3, resulted in shoot elongation. All the elongated shoots were rooted on WPM supplemented with 9.84 μm IBA, and plantlets were transplanted to soil successfully. Chemical names used: 6-benzyladenine (BA), gibberellic acid (GA3), indole-3-butyric acid (IBA), 1-naphthalene acetic acid (NAA), plant growth regulator (PGR), thidiazuron (TDZ), woody plant medium (WPM).
5
Brand, Mark H. "099 Indirect and Direct Regeneration of Kalmia latifolia." HortScience 34, no. 3 (June 1999): 458D—458. http://dx.doi.org/10.21273/hortsci.34.3.458d.
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To introduce desirable trait genes into Kalmia latifolia, efficient adventitious shoot regeneration methods are needed. Silver Dollar (S$) callus induction and growth in the dark was compared on Woody Plant (WP) medium containing 2,4-dichlorophenoxyacetic acid (2,4-D) (1, 5, 10, 20 μM) or naphthaleneacetic acid (NAA) (1, 10, 20, 40 μM) with and without 5 μM isopentenyladenine (2iP). Both 2,4-D and NAA produced >450 mg of callus from leaf explants in 8 weeks. The addition of 2iP tripled growth for 2,4-D and doubled growth for NAA. Greatest callus growth was obtained on 20-40 μM NAA or 5-20 μM 2,4-D. Shoot regeneration on callus was achieved on WP medium containing 30 μM 2iP or 1 μM thidiazuron (TDZ), but a combination of the two was best, with 68% of dark-grown calli regenerating shoots in 4 weeks. 26% more dark-grown calli regenerated shoots than light-grown calli. The type of auxin (2,4-D or NAA) used to grow the calli did not affect shoot regeneration. For direct shoot regeneration, S$ leaf explants were tested on WP medium containing 5, 15, 30, 45 and 60 μM 2iP. The addition of 1 μM indole-3-butyric acid (IBA) doubled the percentage of leaves that regenerated shoots. 2iP concentrations between 15 and 45 μM supported excellent shoot regeneration, but optimal regeneration (95% of explants, 5.1 shoots/leaf) occurred on 30 μM 2iP+1 μM IBA. Leaf explants of six cultivars were grown on optimal medium with shoot regeneration ranging from 17% to 93% of leaves and 1.8 to 8.2 shoots per leaf, depending on the cultivar.
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Orlikowska, Teresa, Agnieszka Marasek, and Danuta Kucharska. "Regeneration of Paeonia mlokosewitschii Lom. and P. tenuifolia L. in vitro from different explants." Acta Societatis Botanicorum Poloniae 67, no. 3-4 (2014): 223–27. http://dx.doi.org/10.5586/asbp.1998.026.
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The pattern of regeneration from tissues of <em>Paeonia mlokosewitschii</em> and <em>P. tenuifolia</em> cultured in vitro in the same chemical conditions depended on the initial explant. Direct shoot regeneration was obtained from the bases of petioles and petals, and leaf veins. Vegetative initial buds and regenerated in vitro shoots produced on their bases slowly growing nodular callus which was very productive in repetitive shoot regeneration. The tops of stems, flower bases, sepals, petals and ovary walls produced small callus which regenerated white and red spherical structures within 1.5 years. After that time also from those cultures arised nodular, shoot regenerating callus developed.
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George, M. W., and R. R. Tripepi. "084 Plant Preservative Mixture (PPM) can Reduce Shoot Regeneration from Leaf Explants of Selected Plants." HortScience 34, no. 3 (June 1999): 455E—455. http://dx.doi.org/10.21273/hortsci.34.3.455e.
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Plant preservative mixture (PPM) is a new broad-spectrum biocide that may be useful for plant tissue culture. The objective of this study was to determine if PPM interfered with adventitious shoot regeneration on leaf explants from several plant species. Leaf explants from Dendranthema grandiflora `Iridon', Betula pendula, Rhododendron catawbiense var. album and R.c. `America' were made from the top two apical leaves on the microshoots. In the first experiment, 0, 0.5, 1, 2, or 4 mL·L-1 PPM were added to species-appropriate regeneration media. In the second experiment, only mum leaf explants were placed on regeneration media containing 0, 0.1, 0.2, 0.3, or 0.4 mL·L-1 PPM. The percentage of explants forming shoots and the number of shoots per regenerating explant were recorded after 4, 6, and 10 weeks, for mum, birch, and rhododendron leaves, respectively. The percentages of shoot regeneration from birch and rhododendron leaf explants were unaffected by up to 4 mL·L-1 PPM, and the number of shoots formed per R.c. album explant were also unaffected by the tested concentrations of PPM. In contrast, the numbers of shoots formed on birch and `America' explants were reduced 48% and 25%, respectively, when 4 mL·L-1 PPM was used in the media. The percentages of shoot regeneration and number of shoots per explant were drastically reduced on mum explants when only 0.5 mL·L-1 PPM was used in the medium. In fact, 0.3 mL·L-1 PPM or higher reduced shoot formation by more than 5-fold. This study demonstrates that the effects of PPM on shoot regeneration from leaf explants are species specific.
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Hebert, Cary J., Darren H. Touchell, Thomas G. Ranney, and Anthony V. LeBude. "In Vitro Shoot Regeneration and Polyploid Induction of Rhododendron ‘Fragrantissimum Improved’." HortScience 45, no. 5 (May 2010): 801–4. http://dx.doi.org/10.21273/hortsci.45.5.801.
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Rhododendron L.‘Fragrantissimum Improved’ is an attractive cultivar with showy, fragrant flowers but has limited potential for breeding because it is a sterile wide hybrid. Protocols for in vitro regeneration and polyploid induction were developed for this cultivar as a means to potentially restore fertility and enhance ornamental traits. Combinations of thidiazuron (TDZ) at 0, 5, 10, 15, or 20 μM and 1-naphthaleneacetic acid (NAA) at 0, 2.5, 5, or 10 μM were used to induce shoot regeneration from leaves. Shoot regeneration was optimized (68% of leaf segments produced shoots) using 8.8 μM TDZ and 10 μM NAA. To induce polyploidy, regenerative callus was treated with 7.5, 15, 30, 60, or 90 μM of the mitotic inhibitor oryzalin for 1, 3, 5, 7, or 14 d in various combinations. Oryzalin significantly affected survival and shoot regenerative capacity. A percentage of homogenous, tetraploid shoots was recovered from treatments of 30 μM oryzalin for 1 (13%) or 3 (13%) days and 7.5 μM oryzalin for 7 (20%) or 14 (7%) days.
9
Cao, Xiaoling, Freddi A. Hammerschlag, and Larry Douglass. "A Two-step Pretreatment Significantly Enhances Shoot Organogenesis from Leaf Explants of Highbush Blueberry cv. Bluecrop." HortScience 37, no. 5 (August 2002): 819–21. http://dx.doi.org/10.21273/hortsci.37.5.819.
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As part of a program to improve highbush blueberry (Vaccinium corymbosum L.) cultivars via tissue culture and genetic engineering, studies were conducted to determine optimum conditions for organogenesis from leaf explants of the previously recalcitrant cv. Bluecrop. The effects of a pretreatment, growth regulators, and age of explant source on shoot organogenesis were investigated. A maximum of 98% explants regenerated shoots with a mean of 11 shoots per leaf explant after 62 days when explants of 2-week-old shoot cultures were incubated on the following regime: pretreatment medium #1 containing 5 μm TDZ and 2.6 μm NAA for 4 days, pretreatment medium #2 containing 7 μm zeatin riboside and 2.6 μm NAA for 3 days, regeneration medium containing 1 μm TDZ for 6 weeks, and last on medium without growth regulators for 10 days. No shoot regeneration occurred if explants were incubated without exposure to pretreatment prior to incubation on regeneration medium. There were no significant differences in percentage of regeneration or the number of shoots regenerating per explant from leaf explants derived from either 1-, 2-, or 3-week-old shoot cultures. Shoot production per explant on regeneration medium containing 1 μm TDZ was about three times that on 0.5 μm TDZ or 20 μm zeatin riboside, and nine times that on 5 μm TDZ. Chemical names used: 1-phenyl-3-(1,2,3-thiadiazol-5-yl)urea (thidiazuron, TDZ); 9-(β-D-ribofuranosyl)-6-(4-hydroxy-3-methyl-but-2-enylamino)purine (zeatin riboside).
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Stamp, James A., Sheila M. Colby, and Carole P. Meredith. "Improved Shoot Organogenesis from Leaves of Grape." Journal of the American Society for Horticultural Science 115, no. 6 (November 1990): 1038–42. http://dx.doi.org/10.21273/jashs.115.6.1038.
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Adventitious shoots developed within 3 weeks from the petiolar stub and, less often, from wounded lamina tissues when leaves excised from nodal cultures of Vitis vinifera L. cvs. French Colombard and Thompson Seedless were cultured on solid Nitsch and Nitsch medium containing BAP at 2 mg·liter-1. The youngest leaf that could be excised, from 1 to 8 mm long, was the most responsive (90% of explants producing shoots compared to 16% for leaf 6). Removal of the lamina from the petiolar stub within the first 3 weeks of culture reduced shoot production. Increase in nodal culture age, without transfer to fresh medium, had no effect on subsequent regeneration from the youngest leaves but did reduce the regeneration frequency of leaves at the next position from 43% to 20%. In regularly subculture nodal cultures, the number of transfers had no effect on subsequent regeneration. Leaves from recently established shoot tip cultures were more responsive than leaves from nodal cultures. The frequency of shoot production was higher in laterally bisected than intact leaves (70% vs. 43%) due to additional regeneration from the distal leaf half at the sites of severed veins. Shoot outgrowth was promoted by the isolation and subculture of regenerating tissue to fresh regeneration medium. Petiolar stub removal promoted de novo shoot organogenesis from the resulting lamina wound. Shoots rooted at a high frequency on Murashige and Skoog medium with 1 mg IA-A/liter and produced morphologically normal plants. Chemical names used: 6-benzylaminopurine (BAP); indole-3-acetic acid (IAA).
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Qu, Luping, James Polashock, and Nicholi Vorsa. "527 A Highly Efficient in Vitro Cranberry Regeneration System using Leaf Explants." HortScience 34, no. 3 (June 1999): 536D—536. http://dx.doi.org/10.21273/hortsci.34.3.536d.
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We have established a very efficient cranberry regeneration (shoot organogenesis) system from leaf explants using a basal medium consisting of Anderson's salts and Murashige and Skoog (MS) organics supplemented with 1-phenyl-3-(1,2,3-thiadiazol-5-yl) urea (TDZ) and N6-(-??-dimethyallylamino) purine) (2ip). Characteristics examined include combinations of varying levels of three plant growth regulators (TDZ, 2ip, and naphthaleneacetic acid (NAA), explant orientation (adaxial or abaxial side in contact with the media), and leaf position relative to the distal end of the shoot. Genotypes (`Early Black', `Pilgrim', `Stevens', `Ben Lear', and US#35) differed significantly in regeneration capacity, and there were no genotype by treatment interaction effects. Regeneration occurred on more than 95% of the explants with `Early Black' and `Pilgrim' producing as many as 100 shoot tips per explant with one particular treatment. Emerging adventitious shoots were always observed on the adaxial side of the leaves regardless of explant orientation. However, regeneration was much greater when the adaxial side was in contact with the media. Regeneration efficiency was not significantly affected by leaf position (10 leaves). Elongation of shoot tips began about 2 weeks after the regenerating explants were transferred to the basal medium without hormones and continued for several months. Elongated shoot cuttings rooted readily.
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Zhang, Yan, and Prem L. Bhalla. "In vitro shoot regeneration from commercial cultivars of Australian canola (Brassica napus L.)." Australian Journal of Agricultural Research 55, no. 7 (2004): 753. http://dx.doi.org/10.1071/ar03209.
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Canola, (Brassica napus L.) is an important crop in Australia. Large genetic variability in the Australian canola cultivars is reflected by their diverse agronomic characteristics. Further improvement using modern breeding methods will lead to the generation of better canola varieties suited for Australian conditions. Genetic engineering relies on the development of efficient methods for regeneration of viable shoots from cultured tissues, and the successful application of transformation techniques. This study reports the in vitro shoot regeneration potential from seedling explants of 7 commercial genotypes (Dunkeld, Grouse, RK7, RI25, Oscar, Rainbow, and Monty) of Australian canola. Seedling explants of these genotypes were all responsive to shoot regeneration. Total number of shoots regenerated varied significantly among the 7 genotypes. Based on the number of shoots regenerated, Rainbow was found to be the most amenable to in vitro regeneration with 55% of cotyledon explants regenerating 2.47 shoots per explants on shoot initiation medium containing 6-benzylaminopurine (3 mg/L), 1-naphthylacetic acid (0.2 mg/L), and gibberellic acid (0.01 mg/L). Normal fertile canola plants from all the 7 genotypes were regenerated. The results obtained from this study will form the basis for genetic transformation studies.
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Kapaun, James A., and Zong-Ming Cheng. "Plant Regeneration from Leaf Tissues of Siberian Elm." HortScience 32, no. 2 (April 1997): 301–3. http://dx.doi.org/10.21273/hortsci.32.2.301.
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Plants were regenerated from leaf tissue of greenhouse-grown seedlings of Siberian elm (Ulmus pumila L.). Shoot regeneration was induced on Murashige and Skoog (MS) medium containing 5 to 10 μm of BA. Up to 55% of the leaf explants formed shoots with an average of 2.4 shoots per explant. Addition of 2.5 or 5 μm of IBA failed to enhance regeneration. Thidiazuron at 0.5 or 1.0 μm also induced shoot regeneration, but the shoots failed to elongate as well as shoots regenerated from media containing BA. Incubation in darkness for 7, 14, or 21 d had little effect in promoting shoot regeneration, except that incubation for 21 d increased shoot regeneration on the medium with 5.0 μm BA. Genotypes differed in shoot regeneration potential, with regeneration frequencies ranging from 13% to 55%. Regenerated shoots were micropropagated on Driver and Kuniyuki Walnut medium. Ninety percent of microcuttings rooted directly in potting soil. This regeneration system will be valuable for genetic transformation and cell selection of Siberian elm. Chemical names used: 6-benzylaminopurine (BA); indole-3-butyric acid (IBA); N-phenyl-N′ -1,2,3-thiadiazol-5-ylurea (thidiazuron, TDZ).
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Sorvari, S., S. Ulvinen, T. Hietaranta, and H. Hiirsalmi. "Preculture Medium Promotes Direct Shoot Regeneration from Micropropagated Strawberry Leaf Disks." HortScience 28, no. 1 (January 1993): 55–57. http://dx.doi.org/10.21273/hortsci.28.1.55.
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The effect of preculturing in vitro plantlets of two strawberry (Fragaria ×ananassa Duch.) cultivars grown on micropropagation medium with and without hormones on regenerating shoots from leaf disks was examined. Preculturing stock plants on micropropagation medium with hormones (BAP at 0.5 mg·liter-1 + IBA at 0.5 mg·liter-1 GA, at 0.2 mg·liter-1) promoted shoot regeneration in the two cultivars tested. Using hormone-containing micropropagation medium for preculture, the highest mean regeneration rate of 9.9 shoots per total number of leaf disks was obtained for the Finnish cultivar Hiku on modified Murashige and Skoog (MS) regeneration medium supplemented with (in mg·liter-1) 2000 KNO3, 400 casein hydrolysate (CH), 3 BAP, and 0.1 IBA. For the Norwegian cultivar Jonsok, the highest mean regeneration rate of 12.8 shoots per total number of leaf disks was obtained on modified MS regeneration medium with (in mg·liter-1) 600 CH, 3 BAP, and 0.1 IBA. Chemical names used: 6-benzylaminopurine (BAP); 3-indolebutyric acid (IBA); gibberellic acid (GA).
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Cao, X., and F. A. Hammerschlag. "Improved Shoot Organogenesis from Leaf Explants of Highbush Blueberry." HortScience 35, no. 5 (August 2000): 945–47. http://dx.doi.org/10.21273/hortsci.35.5.945.
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As part of a program to develop transgenic highbush blueberry (Vaccinium corymbosum L.) cultivars, studies were conducted to determine optimum conditions for high efficiency shoot regeneration from leaf explants of shoots propagated in vitro. The effects on shoot organogenesis of age of explant source, length of dark treatment, the addition of either thidiazuron (TDZ) at 1 or 5 μm, or zeatin riboside at 20 μm to the regeneration medium, and a photosynthetic photon flux (PPF) of either 18 ± 5 or 55 ± 5 μmol·m–2·s–1 were investigated. A maximum of 13.0, 13.0, 12.6, and 4.6 shoots regenerating per explant for cultivars Duke, Georgiagem, Sierra, and Jersey, respectively, occurred on regeneration medium with zeatin riboside and under a PPF of 55 ± 5 μmol·m–2·s–1. `Duke' regenerated equally well on medium with either zeatin riboside or 1 μm TDZ, whereas the number of shoots per explant for `Georgiagem' and `Sierra' was significantly higher on zeatin riboside. Regeneration of `Duke', `Jersey', and `Sierra' on zeatin riboside was significantly better under a PPF of 55 ± 5 μmol·m–2·s–1 than under 18 ± 5 μmol·m–2·s–1, but the higher PPF inhibited regeneration of `Duke' on 5 μm TDZ. There were no significant differences in percentage of regeneration or the number of shoots per explant from leaf explants derived from either 1-, 2-, or 3-week-old shoot cultures, or when either 1 week or 2 weeks of darkness preceded light treatments. Chemical names used: 1-phenyl-3-(1,2,3-thiadiazol-5-yl)urea (thidiazuron, TDZ); 9-(-β-ribofuranosyl)-6-(4-hydroxy-3-methyl-but-2-enylamino)purine (zeatin riboside).
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Miranda, Jacintha, Michele N. Konschuh, E. C. Yeung, and C. C. Chinnappa. "In vitro plantlet regeneration from hypocotyl explants of Stellaria longipes (Caryophyllaceae)." Canadian Journal of Botany 77, no. 2 (July 27, 1999): 318–22. http://dx.doi.org/10.1139/b99-024.
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An in vitro regeneration protocol for Stellaria longipes Goldie was developed using young hypocotyl explants. Optimal regeneration was obtained using Murashige and Skoog (MS) basal medium supplemented with 0.5 µM N6-benzyladenine and 1 µM indole-3-butyric acid. Three different patterns of shoot regeneration were observed: (i) "direct shoot" formation within 3-5 days of inoculation, (ii) nodular structures appeared followed by shoot formation, and (iii) callus formation followed by the appearance of shoots. Histological observation revealed that cells within the central vascular cylinder of the hypocotyl were responsible for shoot organogenesis. Shoot production was not synchronous or uniform among explants. A more synchronous shoot production was obtained by excising the direct shoots or by wounding the nodular structures. Excision and wounding increased the regeneration capability of the explants. Regenerated shoots were readily rooted in MS medium lacking growth regulators and were successfully transferred to greenhouse conditions. These showed morphology consistence with greenhouse-grown plants.Key words: hypocotyl, organogenesis, regeneration, Stellaria longipes.
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Jin, Wanmei, Yuanhua Wang, and Hua Wang. "Adventitious shoot regeneration from leaves of apple rootstock ‘Pingyitiancha’ (Malus hupehensis var. pinyiensis) and genetic fidelity of regenerated plantlets using SSR markers." Canadian Journal of Plant Science 94, no. 8 (November 2014): 1345–54. http://dx.doi.org/10.4141/cjps2013-357.
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Jin, W., Wang, Y. and Wang, H. 2014. Adventitious shoot regeneration from leaves of apple rootstock ‘Pingyitiancha’ (Malus hupehensis var. pinyiensis) and genetic fidelity of regenerated plantlets using SSR markers. Can. J. Plant Sci. 94: 1345–1354. Apple is one of the major fruit tree species in China, its cultivation area and total output rank first in the world. ‘Pingyitiancha’ (Malus hupehensis var. pinyiensis) is a widely used rootstock for apple cultivation in China. Several factors affecting leaf regeneration were investigated. In this study, a successful adventitious shoot regeneration protocol for this cultivar was established. ‘Pingyitiancha’ leaves were a suitable source of explants for regeneration of adventitious shoots. The optimal adventitious shoot regeneration protocol involved subculturing seedling leaves for 30–35 d. The optimum medium was Murashige and Skoog (MS) medium containing 2.0 mg L−1 thidiazuron and 0.2 mg L−1 indole-3-butyric acid. Explants with the abaxial surface in contact with the medium kept for 14 d in the dark showed the highest regeneration percentage of adventitious shoots of explants (100%), and produced an average of 3.6 shoots per regenerating explant. Shoots regenerated from leaves were rooted on half-strength MS medium containing 0.4 mg L−1 1-naphthalene acetic acid. The rooting percentage was 94.4%. Using SSR markers, all banding profiles from regenerated plantlets were monomorphic and same to those of the mother plant. It showed that the uniformity of the in vitro regenerated plantlets was maintained.
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Debnath, Samir C. "A Two-step Procedure for Adventitious Shoot Regeneration from in vitro-derived Lingonberry Leaves: Shoot Induction with TDZ and Shoot Elongation Using Zeatin." HortScience 40, no. 1 (February 2005): 189–92. http://dx.doi.org/10.21273/hortsci.40.1.189.
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The effects of TDZ (0, 0.1, 1, 5 and 10 μm) and explant orientation on adventitious shoot regeneration of `Erntedank' lingonberry were studied. Moderate concentration (1 to 5 μm) of TDZ supported bud and shoot regeneration, but strongly inhibited shoot elongation. TDZ initiated cultures were transferred to medium containing 1-2 μm zeatin and produced usable shoots after one additional subculture. Adventitious bud and shoot regeneration was greatly influenced by explant orientation. Elongated shoots were rooted on a 2 peat: 1 perlite (v/v) medium, and the plantlets were acclimatized and eventually established in the greenhouse with 80% to 90% survival rate.
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Subban, Patharajan, Yaarit Kutsher, Dalia Evenor, Eduard Belausov, Hanita Zemach, Adi Faigenboim, Samuel Bocobza, Michael P. Timko, and Moshe Reuveni. "Shoot Regeneration Is Not a Single Cell Event." Plants 10, no. 1 (December 29, 2020): 58. http://dx.doi.org/10.3390/plants10010058.
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Shoot regeneration is a key tool of modern plant biotechnology. While many researchers use this process empirically, very little is known about the early molecular genetic factors and signaling events that lead to shoot regeneration. Using tobacco as a model system, we found that the inductive events required for shoot regeneration occur in the first 4–5 days following incubation on regeneration medium. Leaf segments placed on regeneration medium did not produce shoots if removed from the medium before four days indicating this time frame is crucial for the induction of shoot regeneration. Leaf segments placed on regeneration medium for longer than five days maintain the capacity to produce shoots when removed from the regeneration medium. Analysis of gene expression during the early days of incubation on regeneration medium revealed many changes occurring with no single expression pattern evident among major gene families previously implicated in developmental processes. For example, expression of Knotted gene family members increased during the induction period, whereas transcription factors from the Wuschel gene family were unaltered during shoot induction. Expression levels of genes involved in cell cycle regulation increased steadily on regeneration medium while expression of NAC genes varied. No obvious possible candidate genes or developmental processes could be identified as a target for the early events (first few days) in the induction of shoot regeneration. On the other hand, observations during the early stages of regeneration pointed out that regeneration does not occur from a single cell but a group of cells. We observed that while cell division starts just as leaf segments are placed on regeneration medium, only a group of cells could become shoot primordia. Still, these primordia are not identifiable during the first days.
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Al-Khayri, Jameel M., Feng H. Huang, Teddy E. Morelock, and Tahani A. Busharar. "Spinach Tissue Culture Improved with Coconut Water." HortScience 27, no. 4 (April 1992): 357–58. http://dx.doi.org/10.21273/hortsci.27.4.357.
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A preliminary study has shown that the addition of 15% (v/v) coconut water (CW) to the culture medium significantly improved callus growth, shoot-regenerative capacity, and shoot growth in leaf disk cultures of spinach (Spinacia oleracea L.). Subsequently, the influence of a range of CW concentrations, 0%, 5%, 10%, 15%, or 20% (v/v), was examined. Callus weight obtained after 5 weeks showed direct relationship to the concentration of CW. This stimulator action was observed in both cultivars tested in this study, `High Pack' and `Baker'. On CW-containing medium, shoot regeneration was expedited to 4 to 5 weeks compared with 8 to 12 weeks on a CW-free medium. Callus of `Baker' induced on a CW-free medium exhibited a significant increase in shoot regeneration frequency when transferred to a regeneration medium enriched with CW, suggesting that the addition of CW to the regeneration medium only is sufficient to achieve improved regeneration.
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WANG, X. H., Q. M. HUANG, L. WANG, and L. Z. WANG. "EFFECT OF SINGLE-WALL CARBON NANOTUBE ON SOYBEAN (GLYCINE MAX) REGENERATION FROM MATURE COTYLEDONARY NODE EXPLANTS." Nano LIFE 02, no. 04 (December 2012): 1250014. http://dx.doi.org/10.1142/s1793984412500146.
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One of the major limitations in producing transgenic soybeans using the agrobacterium-mediated cotyledonary-node method is low regeneration frequency. An improved highly efficient regeneration system of soybean was established herein. Cotyledonary node explants were placed in shoot initiation medium with single-wall carbon nanotube (SWNT) for adventitious shoots regeneration, and adventitious shoots were subcultured in shoot elongation medium with SWNT for shoot elongation and rooting. 40 mg/L SWNT supplemented in shoot initiation medium was found to be the optimal concentration with shoots regeneration frequency significantly increased by 21.5% compared with the control treatment, while for 4 mg/L and 400 mg/L, the increase was 4.6% and 6.5%, respectively. Faster elongation and rooting of adventitious shoots was observed in shoot elongation medium with 40 mg/L SWNT. Soybean plantlet formation frequency within the limited four weeks supplemented with 40 mg/L SWNT reached 48.4% while in the other three treatments: 4 mg/L, 400 mg/L and control, 0% were observed. These results indicate that supplement of SWNT in the soybean medium can efficiently promote adventitious shoots formation frequency, increase plantlet formation frequency and shorten the regeneration period.
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Iwase, Akira, Hirofumi Harashima, Momoko Ikeuchi, Bart Rymen, Mariko Ohnuma, Shinichiro Komaki, Kengo Morohashi, et al. "WIND1 Promotes Shoot Regeneration through Transcriptional Activation of ENHANCER OF SHOOT REGENERATION1 in Arabidopsis." Plant Cell 29, no. 1 (December 23, 2016): 54–69. http://dx.doi.org/10.1105/tpc.16.00623.
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Faize, Mohamed, Lydia Faize, Lorenzo Burgos, Alan T. Critchley, and Nuria Albuquerque. "Application of Ascophyllum nodosum-Based Soluble Extract on Micropropagation and Regeneration of Nicotiana benthamiana and Prunus domestica." Plants 10, no. 7 (July 2, 2021): 1354. http://dx.doi.org/10.3390/plants10071354.
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In the present study, the effect of a commercial extract of the seaweed Ascophyllum nodosum on in vitro micropropagation, shoot regeneration, and rhizoghenesis were studied in Nicotiana benthamiana and Prunus domestica. Results showed that the MS medium supplemented with various concentrations of the Ascophyllum extract (5, 10, 50, and 100 mg L−1) significantly enhanced the number of regenerated buds from N. benthamiana leaf discs to the conventional MS regenerating medium. Increases ranged from 3.5 to 6.5 times higher than the control. The effect of the Ascophyllum extract on N. benthamiana micropropagation was assessed through the measurement of some plant growth parameters. Results showed that the extract alone could not replace the micropropagation medium since shoot length, shoot diameter, root length, and leaf area were significantly reduced. However, its combination with a half-strength MS medium enhanced these parameters. Its effect was also evaluated on regeneration from plum hypocotyl slices. When added to the shoot regeneration medium without any plant growth regulators, the Ascophyllum extract alone could induce shoot regeneration. However, the percentage of bud regeneration and number of regenerated buds were lower than with the conventional shoot regeneration medium containing complete growth regulators. In contrast, the Ascophyllum extract drastically promoted rhizogenesis from plum hypocotyl slices. These results pave the way for the possible use of A. nodosum extracts in in vitro mass propagation of higher plants.
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Cao, Xiaoling, and F. A. Hammerschlag. "112 The Influence of Plant Growth Regulators and Light Levels on Shoot Morphogenesis from Leaf Explants of Highbush Blueberry." HortScience 34, no. 3 (June 1999): 460F—461. http://dx.doi.org/10.21273/hortsci.34.3.460f.
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As part of a program to develop transgenic highbush blueberry (Vaccinium corymbosum L.) cultivars, studies were conducted to determine optimum conditions for high efficiency shoot regeneration from leaf explants of in vitro-propagated shoot cultures. The effect of either thidiazuron at 1 or 5 μM, or zeatin riboside at 20 μM, and two lit levels (18 ± 5 or 55 ± 5 μmol·m-2·s-1) on shoot organogenesis were investigated. With the exception of `Bluecrop', which did not regenerate shoots, maximum shoot regeneration of 13, 12.7, 12.6 and 4.6 shoots per explant for cultivars Duke, Georgiagem, Sierra, and Jersey, respectively, occured on regeneration medium with zeatin riboside and under a light intensity of 55 μmol·m-2·s-1. Whereas `Duke' regenerated equally well on regeneration medium with either zeatin riboside or 5 μM thidiazuron, regeneration frequencies for `Georgiagem' and `Sierra' were significantly higher on zeatin riboside. A light intensity of 55 μmol·m-2·s-1 significantly increased regeneration of cultivars Duke, Jersey, and Sierra on zeatin riboside, but inhibited regeneration of Duke on 5 μM thidazuron.
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Raspor, Martin, Václav Motyka, Abdul Rasheed Kaleri, Slavica Ninković, Ljiljana Tubić, Aleksandar Cingel, and Tatjana Ćosić. "Integrating the Roles for Cytokinin and Auxin in De Novo Shoot Organogenesis: From Hormone Uptake to Signaling Outputs." International Journal of Molecular Sciences 22, no. 16 (August 9, 2021): 8554. http://dx.doi.org/10.3390/ijms22168554.
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De novo shoot organogenesis (DNSO) is a procedure commonly used for the in vitro regeneration of shoots from a variety of plant tissues. Shoot regeneration occurs on nutrient media supplemented with the plant hormones cytokinin (CK) and auxin, which play essential roles in this process, and genes involved in their signaling cascades act as master regulators of the different phases of shoot regeneration. In the last 20 years, the genetic regulation of DNSO has been characterized in detail. However, as of today, the CK and auxin signaling events associated with shoot regeneration are often interpreted as a consequence of these hormones simply being present in the regeneration media, whereas the roles for their prior uptake and transport into the cultivated plant tissues are generally overlooked. Additionally, sucrose, commonly added to the regeneration media as a carbon source, plays a signaling role and has been recently shown to interact with CK and auxin and to affect the efficiency of shoot regeneration. In this review, we provide an integrative interpretation of the roles for CK and auxin in the process of DNSO, adding emphasis on their uptake from the regeneration media and their interaction with sucrose present in the media to their complex signaling outputs that mediate shoot regeneration.
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Fu, Dong, Tan, Yin, Zhang, Zhao, Ye, and Wu. "Identification of Shoot Differentiation-Related Genes in Populus euphratica Oliv." Genes 10, no. 12 (December 11, 2019): 1034. http://dx.doi.org/10.3390/genes10121034.
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De novo shoot regeneration is one of the important manifestations of cell totipotency in organogenesis, which reflects a survival strategy organism evolved when facing natural selection. Compared with tissue regeneration, and somatic embryogenesis, de novo shoot regeneration denotes a shoot regeneration process directly from detatched or injured tissues of plant. Studies on plant shoot regeneration had identified key genes mediating shoot regeneration. However, knowledge was derived from Arabidopsis; the regeneration capacity is hugely distinct among species. To achieve a comprehensive understanding of the shoot regeneration mechanism from tree species, we select four genetic lines of Populus euphratica from a natural population to be sequenced at transcriptome level. On the basis of the large difference of differentiation capacity, between the highly differentiated (HD) and low differentiated (LD) groups, the analysis of differential expression identified 4920 differentially expressed genes (DEGs), which were revealed in five groups of expression patterns by clustering analysis. Enrichment showed crucial pathways involved in regulation of regeneration difference, including “plant hormone signal transduction”, “cell differentiation”, "cellular response to auxin stimulus", and “auxin-activated signaling pathway”. The expression of nine genes reported to be associated with shoot regeneration was validated using quantitative real-time PCR (qRT-PCR). For the specificity of regeneration mechanism with P. euphratica, large amount of DEGs involved in "plant-pathogen interaction", ubiquitin-26S proteosome mediated proteolysis pathway, stress-responsive DEGs, and senescence-associated DEGs were summarized to possibly account for the differentiation difference with distinct genotypes of P. euphratica. The result in this study helps screening of key regulators in mediating the shoot differentiation. The transcriptomic characteristic in P. euphratica further enhances our understanding of key processes affecting the regeneration capacity of de novo shoots among distinct species.
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Smith, Nicole A., and Prem L. Bhalla. "Comparison of shoot regeneration potential from seedling explants of Australian cauliflower (Brassica oleracea var. botrytis) varieties." Australian Journal of Agricultural Research 49, no. 8 (1998): 1261. http://dx.doi.org/10.1071/a98067.
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Genetic engineering of crop plants relies on the development of efficient methods for the regeneration of viable shoots from cultured tissues. The objective of the present study was to develop a protocol for efficient shoot and plant regeneration from seedling explants of commercial cauliflower (B. oleracea var. botrytis) genotypes and to compare the regeneration capacity of the most commonly used explants: cotyledon, hypocotyl, and root. A combination of growth hormones including 6-benzylaminopurine, 1-naphthylacetic acid, and gibberellic acid was used in the MS-based medium, and factors influencing regeneration of shoots were investigated. Using the protocol described here, shoots from hypocotyl, cotyledon, and root explants of all the 11 genotypes tested were able to be regenerated. Root and hypocotyl explants produced more callus than cotyledon explants and also were more responsive to shoot regeneration, as a high percentage (>71% and >80% for hypocotyl and roots, respectively) of shoot initiation from these explants was observed. In addition, root and hypocotyl explants also produced more shoots per explant than cotyledon explants. The in vitro regenerated shoots were successfully rooted and acclimatised to glasshouse conditions. This study shows that seedling explants of cauliflower are amenable to multiple shoot formation with high regeneration frequencies, and could be used for genetic transformation experiments.
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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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Matsuo, Naoki, Hiromi Mase, Miho Makino, Hiro Takahashi, and Hiroharu Banno. "Identification of ENHANCER OF SHOOT REGENERATION 1-upregulated genes during in vitro shoot regeneration." Plant Biotechnology 26, no. 4 (2009): 385–93. http://dx.doi.org/10.5511/plantbiotechnology.26.385.
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Swanberg, Andrea, and Wenhao Dai. "Plant Regeneration of Periwinkle (Catharanthus roseus) via Organogenesis." HortScience 43, no. 3 (June 2008): 832–36. http://dx.doi.org/10.21273/hortsci.43.3.832.
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Two periwinkle cultivars, Pacific Coral (P1) and Sunstorm Rose (P2), were used for development of a plant regeneration system. Leaf and internodal explants collected from in vitro plants were plated onto woody plant medium (WPM) using a factorial arrangement of 6-benzyladeine (BA) and 1-naphthalene acetic acid (NAA). Shoots were successfully regenerated. Shoot production from leaf tissues was minimal for all cultivars, whereas internodal tissues showed variable rates of regeneration depending on the hormone combination. Cultivar P1 showed the maximum regeneration rate (73.3%) when internodal explants, 4 to 6 mm in length, were placed on WPM containing 5 μm BA and 5 μm NAA. Cultivar P2 showed a regeneration rate of 56.7% with a combination of 20 μm BA and 10 μm NAA. Shoot regeneration rate increased as the internodal explant size increased for P2; however, the regeneration rate decreased when the explant size was greater than 7 mm for P1. The shoot regeneration rate decreased as the period of the dark treatment of internodal explants increased in both P1 and P2. The antibiotics carbenicillin (Carb) and cefotaxime (Cef) had little effect on shoot regeneration. There was a slightly higher rate observed for P1 when Cef was added into the medium, whereas P2 showed a decrease with the addition of Cef. Carb showed no significant effect on shoot regeneration for both cultivars. Addition of both Carb and Cef to the medium slightly inhibited shoot regeneration.
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Debnath, Samir C., and Danny L. Barney. "Shoot Regeneration and Plantlet Formation by Cascade Huckleberry, Mountain Huckleberry, and Oval-leaf Bilberry on a Zeatin-containing Nutrient Medium." HortTechnology 22, no. 1 (February 2012): 106–13. http://dx.doi.org/10.21273/horttech.22.1.106.
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A plant regeneration protocol was developed for cascade huckleberry (Vaccinium deliciosum), mountain huckleberry (V. membranaceum), and oval-leaf bilberry (V. ovalifolium) clones. The effects of zeatin concentrations (0, 4.6, 9.1, and 13.7 μM) and explant type (leaf or stem segment) on adventitious shoot regeneration were studied on a nutrient medium of low ionic concentration. Adventitious bud and shoot regeneration was greatly influenced by clone, explant type, and zeatin concentration. Zeatin at 9.1 to 13.7 μM supported the best bud and shoot regeneration. At low concentrations (2.3 to 4.6 μM), zeatin enhanced shoot elongation and produced usable shoots after one additional subculture. The three clones differed significantly with respect to multiplication rate of adventitious shoots. Oval-leaf bilberry and mountain huckleberry clones produced six to seven 5-cm-long shoots per explant and cascade huckleberry clone produced five 3-cm-long shoots per explant, when 2.3 μM zeatin was used in the medium. Increasing the concentration of zeatin in the culture medium increased shoot number per explant, but decreased shoot height, leaf number per shoot, and shoot vigor. Proliferated shoots were rooted on the same medium but without any plant growth regulators (PGRs). Rooted plantlets were transferred to a 2 peat:1 perlite (v/v) medium for acclimatization and eventually established in the greenhouse with 75% to 90% survival rate. This in vitro protocol will be useful for micropropagation, in vitro selection, and genetic manipulation of Vaccinium species.
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Ricci, Angela, Luca Capriotti, Bruno Mezzetti, Oriano Navacchi, and Silvia Sabbadini. "Adventitious Shoot Regeneration from In Vitro Leaf Explants of the Peach Rootstock Hansen 536." Plants 9, no. 6 (June 16, 2020): 755. http://dx.doi.org/10.3390/plants9060755.
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In the present study, an efficient system for the in vitro regeneration of adventitious shoots from the peach rootstock Hansen 536 leaves has been established. Twenty regeneration media containing McCown Woody Plant Medium (WPM) as a basal salt supplemented with different concentrations and combinations of plant growth regulators (PGRs) were tested. Expanded leaves along with their petiole from 3-week-old elongated in vitro shoot cultures were used as starting explants. The highest regeneration rate (up to 53%) was obtained on WPM basal medium enriched with 15.5 μM N6-benzylaminopurine (BAP). The influences on leaf regeneration of the ethylene inhibitor silver thiosulphate (STS) and of different combinations of antibiotics added to the optimized regeneration medium were also investigated. The use of 10 μM STS or carbenicillin (238 μM) combined with cefotaxime (210 μM) significantly increased the average number of regenerating shoots per leaf compared to the control. In vitro shoots were finally elongated, rooted and successfully acclimatized in the greenhouse. The results achieved in this study advances the knowledge on factors affecting leaf organogenesis in Prunus spp., and the regeneration protocol described looks promising for the optimization of new genetic transformation procedures in Hansen 536 and other peach rootstocks and cultivars.
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Vyapari, Sudeep, and Houchang Khatamian. "586 PB 123 AXILLARY SHOOT REGENERATION IN CHINKAPIN OAK." HortScience 29, no. 5 (May 1994): 515g—515. http://dx.doi.org/10.21273/hortsci.29.5.515g.
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Surface disinfested nodal and shoot-tip sections of chinkapin oak (Quercus muehlenbergii Engelm.), obtained from adult or juvenile source, when cultured on WFM supplemented with BA or kinetin (1.0 -5.0 mg l-1) produced greater number of axillary shoots per explant and shoot lengths than MS medium. Nodal and shoot-tip explants cultured in WPM containing cytokinins, BA or kinetin (0.1 - 5.0 mg l“) resulted in greater number of axillary shoots than media containing auxins, 2,4-D or NAA (1.0 - 5.0 mg l-1). In vitro grown shoot explants cultured in WFM shoot multiplication medium containing thidiazuron did not produce axillary shoots. Microshoots when cultured in WFM plus NAA or IBA (0.1 -2.0 mg l-1), or subjected to IBA (0.5 mg l-1) pulse treatment (0, 5, 10 or 15 min.) did not root.
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Barik, Durga Prasad, Umaballava Mohapatra, and Pradeep Kumar Chand. "Direct shoot regeneration from epicotyl explants of grasspea (Lathyrus sativus)." Australian Journal of Botany 54, no. 5 (2006): 505. http://dx.doi.org/10.1071/bt05152.
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A reproducible procedure is described for adventitious shoot organogenesis in epicotyl segments resulting in prolific plant regeneration of a grain legume grasspea (Lathyrus sativus L.). Among seedling explant types examined, epicotyl segments were most responsive. The highest percentage of direct shoot regeneration was elicited on Murashige–Skoog (MS) medium augmented with 4.0 mg L–1 6-benzyladenine (BA) + 2.0 mg L–1 α-naphthaleneacetic acid (NAA). Compared with four other genotypes examined, IC-120487 showed the highest shoot regeneration frequency (approximately 80%) with maximum shoot numbers (averaging eight shoots per explant) and longest average shoot length (approximately 4 cm). Rhizogenesis was induced in ~78% of the regenerated shoots in half-strength MS medium containing 0.5 mg L–1 indole-3-acetic acid (IAA). Plantlets were acclimated in vermi-compost and 75% of those transferred to soil survived and set viable seeds.
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Habas, Rebaz Rasul, Musa Turker, and Fethi Ahmet Ozdemir. "In vitro Multiple Shoot Regeneration from Petunia hybrida." Turkish Journal of Agriculture - Food Science and Technology 7, no. 10 (October 12, 2019): 1554. http://dx.doi.org/10.24925/turjaf.v7i10.1554-1560.2570.
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An efficient plant regeneration protocol was developed from in vitro germinated seeds of Petunia hybrida an ornamentally important plant in the family Solanaceae. Shoot tip and node explants of Petunia hybrida were cultured on MS basal medium supplemented with different concentrations and combinations of Benzyl amino purine (BAP), 1-Naphthaleneacetic acid (NAA), Indole-3-butyric acid (IBA) and Gibberellic acid (GA3). The highest shoot length was obtained from MS medium supplemented with 1 mg/l BAP + 1 mg/l NAA. The highest shoot number (3 shoots/explant) were obtained from MS medium supplemented with 0.6 mg/l BAP + 0.5 mg/l IBA. The isolated shoots were transferred to MS basal medium supplemented with different concentrations of GA3 ranging from 0.05, 0.2, 0.5 and 1 mg/l for shoot elongation. The highest shoot length (5.75 cm) was recorded from the MS medium supplemented with 0.2 mg/l GA3 +0.2 mg/l BAP. Rooting of regenerated shoots were achieved on MS medium supplemented with 0.1-1 mg/1 IBA and NAA. The regenerated shoots with well developed roots were successfully acclimatized and established in pots containing sterilized peat moss and grown under laboratory conditions with 70% survival rates.
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Vejsadová, H. "Growth regulator effect on in vitro regeneration of rhododendron cultivars." Horticultural Science 35, No. 2 (June 24, 2008): 90–94. http://dx.doi.org/10.17221/643-hortsci.
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In Rhododendron L. cv. Azuro, Bohumil Kavka, Catharine van Toll, Grandiflorum, Mars, Nova Zembla, Ortrud, Ovation, Prof. Scholz, Purple Splendour, Rebe and Van Werden Poelman, the effect of growth regulators on organogenesis induction of shoot-tip meristems was tested. All cultivars significantly showed the highest shoot regeneration on MS medium containing 6 mg/dm<sup>3</sup> isopentenyladenine (2iP). For most rhododendrons, the highest shoot multiplication was found on a medium with 8–10 mg/dm<sup>3</sup> 2iP in combination with 1 mg/dm<sup>3</sup> indoleacetic acid (IAA). Shoots rooted successfully in the substrate with high level of peat without growth regulators. However, the commercial preparation Racine significantly increased rooting in cv. Grandiflorum, Nova Zembla and Rebe compared with 0.03% indolebutyric acid (IBA).
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Narasimhulu, S. B., Shyam Prakash, V. L. Chopra, and V. Arunachalam. "Comparative shoot regeneration in diploid and amphidiploid Brassica species and their interspecific hybrids." Canadian Journal of Botany 70, no. 7 (July 1, 1992): 1513–14. http://dx.doi.org/10.1139/b92-189.
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Shoot regeneration response in interspecific hybrids of Brassica species were assessed in relation to the diploid pollen parents and amphidiploid female parents. Superior regeneration responses were observed in interspecific combinations of B. carinata and B. nigra (BBC), B. juncea and B. campestris (AAB), and B. napus and B. campestris (AAC). Though synthetic B. napus regenerated with a frequency less than that of the better regenerating parent (B. oleracea), higher regeneration response was observed in the hybrid between B. napus and B. campestris. Two triploid combinations of the genetic constitution ABC, one obtained by crossing synthetic B. napus with B. nigra and the other by crossing natural B. juncea with B. oleracea, showed low regeneration responses. The response improved substantially in a tetraploid of the constitution ABBC obtained by crossing B. juncea with B. carinata. Key words: Brassica, alloploids, shoot regeneration.
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Yang, Guochen, and Marihelen Kamp-Glass. "Direct Shoot Organogenesis of Medicago sativa." HortScience 31, no. 4 (August 1996): 628f—628. http://dx.doi.org/10.21273/hortsci.31.4.628f.
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An efficient and reliable protocol of in vitro shoot regeneration must be first established to have a successful genetic transformation. As a member of legume family, alfalfa is very difficult for direct shoot regeneration. There is no published information on direct shoot organogenesis, although success has been well documented on embryogenesis, which must go through callus stage. Different plant growth regulators at various concentrations were evaluated for callus initiation, development, and direct shoot regeneration. Multiple shoots were produced directly from each individual explant. This will provide an efficient means for production of transgenic alfalfa plants. Therefore, genetic transformation of Medicago germplasm will be significantly expedited.
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Xin, W., Z. Liu, Y. Song, T. Hou, and F. Xiang. "Direct shoot regeneration from Arabidopsis thaliana shoot apical meristems." Biologia plantarum 56, no. 4 (December 1, 2012): 601–6. http://dx.doi.org/10.1007/s10535-012-0127-x.
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Malek, MA, MA Bari Miah, M. AL-Amin, D. Khanam, and M. Khatun. "In vitro regeneration in pointed gourd." Bangladesh Journal of Agricultural Research 32, no. 3 (January 8, 2008): 461–71. http://dx.doi.org/10.3329/bjar.v32i3.548.
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An efficient protocol was developed for plant regeneration, multiplication and rooting under in vitro condition in pointed gourd. Highest percent of shoot regeneration was 93.86 when nodal explants were cultured on MS+2.0 mg/1 BAP. The maximum number of shoots (4.00) per explant was observed in MS + 2.0 mg/1 BAP + 0.3 mg/1 NAA from nodal segment. Among the two explants, nodal segment was found better for shoot regeneration. Female genotypes responded better than the male genotypes for shoot induction and proliferation. Lower nodal segment performed the best shoot regeneration. The best response towards root induction was achieved on half MS medium supplemented with 0.5 mg/1 NAA. The regenerated plantlets were successfully established in prepared earthen soil pot.DOI: http://dx.doi.org/10.3329/bjar.v32i3.548Bangladesh J. Agril. Res. 32(3) : 461-471, September 2007
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Kochanová, Zuzana, Naci Onus, and Ján Brindza. "Adventitious shoot regeneration from dormant buds of persimmon (Diospyros kaki Thunb.) cv. Hachiya." Journal of Agrobiology 28, no. 2 (January 1, 2011): 113–18. http://dx.doi.org/10.2478/v10146-011-0012-9.
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Adventitious shoot regeneration from dormant buds of persimmon (Diospyros kakiThunb.) cv. HachiyaThe effects of plant hormones 6-benzylaminopurine (BAP) and indole-3-butyric acid (IBA) on adventitious shoot regeneration from dormant persimmon buds were studied. The object of the study was the persimmon cultivar Hachiya, one of the most important persimmon cultivars in the Mediterranean area and Asia. Shoot regeneration was evaluated 4, 6 and 8 weeks after initiating the hormone treatment. Average shoot length was measured after 8 weeks and was evaluated by LSD test. Except for the media without hormone supplement, there was a statistically significant difference among average values of shoot length of plants, grown on the tested media. The increase in BAP had an effect on shoot regeneration that was significant and more pronounced with the addition of IBA, especially to the MS (1/2 N) medium. The highest value of shoot regeneration (98%) was obtained on medium MS (1/2 N), supplemented with 5 μmol l-1BAP and 1 μmol l-1IBA, with the highest average shoot length 23.69 mm, measured 8 weeks after the experiment initiation. The results indicate that adventitious shoots can be successfully produced in persimmon cv. Hachiya, especially with the supplement of hormone BAP, which, according to our results, plays an important role in persimmonin vitroregeneration.
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Mitić, Nevena, Mariana Stanišić, Jelena Milojević, Ljiljana Tubić, Tatjana Ćosić, Radomirka Nikolić, Slavica Ninković, and Rade Miletić. "Optimization of In Vitro Regeneration from Leaf Explants of Apple Cultivars Golden Delicious and Melrose." HortScience 47, no. 8 (August 2012): 1117–22. http://dx.doi.org/10.21273/hortsci.47.8.1117.
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An efficient in vitro shoot regeneration method from leaf explants of apple cultivars Golden Delicious and Melrose by optimization of regeneration medium, explant type and orientation, dark pre-treatment, and gelling agent is presented. Murashige and Skoog’s regeneration medium containing 22 μM thidiazuron (TDZ) and 1.5 μM indole-3-butyric acid (IBA) (M2 medium) was superior for regeneration as well as for subsequent shoot multiplication in both cultivars, providing regeneration frequency of 95% or higher in the best combination with other factors. Pre-incubation in the dark proved to be an essential factor for regeneration. The use of agar as a gelling agent provides satisfactory regeneration frequency compared with media gelled with PhytagelTM. Leaf explants of cv. Melrose with adaxial surface in contact with M2 medium and those of cv. Golden Delicious orientated contrary regenerated the highest mean number of shoots per explant. Under optimal conditions, a maximal index of shoot-forming capacity of 11.44 and 6.30 for ‘Melrose’ and ‘Golden Delicious’, respectively, was achieved. Regenerated shoots were successfully rooted and acclimated ex vitro.
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Kupriyanova, E. V., E. R. Denisova, M. A. Baier, and T. A. Ezhova. "Differences in the Manifestation of Cell Pluripotence In Vivo and In Vitro in the Mutant Arabidopsis thaliana with the Phenotype of Cell Memory Disorder." Russian Journal of Plant Physiology 68, no. 1 (January 2021): 46–55. http://dx.doi.org/10.1134/s1021443721010106.
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AbstractPlant cells cultivated in vitro are a convenient model for studying the genetic and physiological mechanisms necessary for the cells to acquire a state of pluripotency. Earlier studies on a model plantArabidopsis thaliana(L.) Heynh. have identified the key role of genes that determine the pluripotency of cells in the shoot apical meristem in de novo shoot regeneration in tissue culture. In accordance with this, cells of mutant plants with a higher level of expression of pluripotency genes were characterized by an increased potential for de novo shoot regeneration. Thetaemutant was the exception to this rule. The mutant resumed the expression of pluripotency genes and cell proliferation at the late stages of leaf development, which indicates a violation of the mechanisms for maintaining epigenetic cellular memory. At the same time, leaf cells cultured in vitro showed a lower proliferative activity compared to the wild type and were not capable of de novo regeneration of shoots. A decrease in the regenerative potential of cultured cells of thetaemutant indicates an important role of epigenetic memory in the response of cells to exogenous hormones. Impaired epigenetic memory of leaf cells of thetae mutant and differences in their proliferative and regenerative capacities in planta and in vitro make this mutant a unique model for studying the role of epigenetic modifications in the regulation of cell pluripotency.
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Chiam, Nyet-Cheng, Tomoyo Fujimura, Ryosuke Sano, Nobuhiro Akiyoshi, Ryoko Hiroyama, Yuichiro Watanabe, Hiroyasu Motose, Taku Demura, and Misato Ohtani. "Nonsense-Mediated mRNA Decay Deficiency Affects the Auxin Response and Shoot Regeneration in Arabidopsis." Plant and Cell Physiology 60, no. 9 (August 6, 2019): 2000–2014. http://dx.doi.org/10.1093/pcp/pcz154.
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Abstract Plants generally possess a strong ability to regenerate organs; for example, in tissue culture, shoots can regenerate from callus, a clump of actively proliferating, undifferentiated cells. Processing of pre-mRNA and ribosomal RNAs is important for callus formation and shoot regeneration. However, our knowledge of the roles of RNA quality control via the nonsense-mediated mRNA decay (NMD) pathway in shoot regeneration is limited. Here, we examined the shoot regeneration phenotypes of the low-beta-amylase1 (lba1)/upstream frame shift1-1 (upf1-1) and upf3-1 mutants, in which the core NMD components UPF1 and UPF3 are defective. These mutants formed callus from hypocotyl explants normally, but this callus behaved abnormally during shoot regeneration: the mutant callus generated numerous adventitious root structures instead of adventitious shoots in an auxin-dependent manner. Quantitative RT-PCR and microarray analyses showed that the upf mutations had widespread effects during culture on shoot-induction medium. In particular, the expression patterns of early auxin response genes, including those encoding AUXIN/INDOLE ACETIC ACID (AUX/IAA) family members, were significantly affected in the upf mutants. Also, the upregulation of shoot apical meristem-related transcription factor genes, such as CUP-SHAPED COTYLEDON1 (CUC1) and CUC2, was inhibited in the mutants. Taken together, these results indicate that NMD-mediated transcriptomic regulation modulates the auxin response in plants and thus plays crucial roles in the early stages of shoot regeneration.
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Meyer, Elisabeth M., Darren H. Touchell, and Thomas G. Ranney. "In Vitro Shoot Regeneration and Polyploid Induction from Leaves of Hypericum Species." HortScience 44, no. 7 (December 2009): 1957–61. http://dx.doi.org/10.21273/hortsci.44.7.1957.
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Hypericum L. H2003-004-016 is a complex hybrid among Hypericum frondosum Michx., Hypericum galioides Lam., and Hypericum kalmianum L. and exhibits valuable ornamental characteristics, including compact habit, bluish green foliage, and showy flowers. Inducing polyploidy may further enhance the ornamental traits of this hybrid and provide new opportunities for hybridizing with other naturally occurring polyploid Hypericum sp. In this study, in vitro shoot regeneration and treatment of regenerative callus with the dinitroaniline herbicide oryzalin (3,5-dinitro-N4,N4-dipropylsufanilamide) were investigated as a means of inducing allopolyploidy. First, in vitro regeneration was optimized for callus and shoot induction by culture of leaf explants on medium supplemented with benzylamino purine (BA) or meta-topolin (mT) at 5, 10, or 15 μM in combination with indoleacetic acid (IAA) at 0, 1.25, 2.5, or 5 μM. Both BA and mT treatments successfully induced regenerative callus and shoots. Multiple regression analysis estimated maximum regenerative callus (94%) and shoot induction (18 shoots per explant) in medium supplemented with 5 μM BA and 3.75 μM IAA. In the second part of the study, exposure of regenerative callus to oryzalin at 0, 7.5, 15, 30, 60, or 90 μM for durations of 3, 6, or 9 d was investigated for polyploid induction. There was no survival for any of the calli in the 60- or 90-μM oryzalin treatments, but calli subjected to the other treatments exhibited some survival and polyploid induction. Duration had no effect on callus survival or ploidy level, but oryzalin concentration was a significant factor in both. The greatest percentage (44%) of polyploids was induced with 30 μM oryzalin. Spontaneous chromosome doubling was observed in 8% of control explants receiving no oryzalin treatment.
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Yang, Woorim, Myung-Hwan Choi, Bosl Noh, and Yoo-Sun Noh. "De Novo Shoot Regeneration Controlled by HEN1 and TCP3/4 in Arabidopsis." Plant and Cell Physiology 61, no. 9 (June 24, 2020): 1600–1613. http://dx.doi.org/10.1093/pcp/pcaa083.
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Abstract Plants have the ability to regenerate whole plant body parts, including shoots and roots, in vitro from callus derived from a variety of tissues. However, the underlying mechanisms for this de novo organogenesis, which is based on the totipotency of callus cells, are poorly understood. Here, we report that a microRNA (miRNA)-mediated posttranscriptional regulation plays an important role in de novo shoot regeneration. We found that mutations in HUA ENHANCER 1 (HEN1), a gene encoding a small RNA methyltransferase, cause cytokinin-related defects in de novo shoot regeneration. A hen1 mutation caused a large reduction in the miRNA319 (miR319) level and a subsequent increase in its known target (TCP3 and TCP4) transcript levels. TCP transcription factors redundantly inhibited shoot regeneration and directly activated the expression of a negative regulator of cytokinin response ARABIDOPSIS THALIANA RESPONSE REGULATOR 16 (ARR16). A tcp4 mutation at least partly rescued the shoot-regeneration defect and derepression of ARR16 in hen1. These findings demonstrate that the miR319-TCP3/4-ARR16 axis controls de novo shoot regeneration by modulating cytokinin responses.
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Shibli, Rida A., and M. A. L. Smith. "Direct Shoot Regeneration from Vaccinium pahalae (Ohelo) and V. myrtillus (Bilberry) Leaf Explants." HortScience 31, no. 7 (December 1996): 1225–28. http://dx.doi.org/10.21273/hortsci.31.7.1225.
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Ohelo (V. pahalae Skottsb.) and bilberry (V. myrtillus L.) shoots were regenerated via direct organogenesis from whole leaves and leaf sections and also from hypocotyl explants of bilberry. Explants preincubated for 1 to 2 weeks in darkness yielded ≈75% regeneration frequencies and the highest number of regenerating shoots/explant on TDZ-supplemented media (0.9 to 2.7 μm). When 2iP or zeatin were substituted as the cytokinin source, frequencies of regeneration and shoot productivity were significantly lower. Explants held under constant illumination (no dark pretreatment) had significantly lower regeneration frequencies in all tested cytokinin-supplemented media. 2,4-D stimulated callus formation, but did not support regeneration from vegetative explants. Cells from callus and suspension cultures did not exhibit regeneration in any of the media that supported organogenesis from leaves. Regenerants were successfully micropropagated, although callus formation caused by zeatin and high 2iP levels interfered with shoot proliferation. Zeatin induced hyperhydricity in shoots from both species, but more severely in ohelo. Ex vitro rooting after treatment with 4.9 μm IBA or 5.4 μm NAA was 95% and 60% successful for bilberry and ohelo, respectively, and plants were readily acclimatized after an interval in a fog chamber. Bilberry microshoots also rooted in vitro in the absence of growth regulator treatment. Chemical names used: 1H-indole-3-butanoic acid (IBA); N-(3-methyl-2-butenyl)-1-H-purine-6-amine (2iP); 6-furfurylaminopurine (kinetin); 1-naphthaleneacetic acid (NAA); thidiazuron=1-phenyl-3-(1,2,3-thiadiazio-5-yl)urea (TDZ); 2,4-dichlorophenoxyacetic acid (2,4-D); 6-(4-hydroxy-3-methylbut-2-enylamino) purine (zeatin).
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Matsuo, Naoki, and Hiroharu Banno. "Arabidopsis ENHANCER OF SHOOT REGENERATION 2 and PINOID are involved in in vitro shoot regeneration." Plant Biotechnology 29, no. 4 (2012): 367–72. http://dx.doi.org/10.5511/plantbiotechnology.12.0514a.
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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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Pykalo, S. V., O. A. Demydov, T. V. Yurchenko, N. I. Prokopik, and O. V. Humeniuk. "The regeneration potential of promising winter common wheat lines in shoot apical meristem culture." Faktori eksperimental'noi evolucii organizmiv 25 (August 30, 2019): 298–303. http://dx.doi.org/10.7124/feeo.v25.1181.
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Aim. To investigate the regenerative ability of promising winter common wheat lines in shoot apical meristem culture. Methods. Plant tissue culture methods, statistical evaluation of data. Results. The processes of morphogenesis in culture of apical meristem of 3-days seedlings of lines of winter common wheat were investigated and it was established that the frequency of callusogenesis and shoot regeneration in the studied forms is determined by the genotype of explant. Two types of callus with morphophysiological properties were identified: morphogenic and nonmorphogenic callus. The formation of regenerated plants from wheat calli took place through both gemmorizogenesis and somatic embryogenesis. Conclusions. The line Erytrospermum 60068 was characterized the highest regeneration potential and it can be recommended for further biotechnology of wheat. Obtained technology of vigorous regenerated plant production of winter common wheat lines in shoot apical meristem culture can be used in cell selection and genetic engineering experiments. Keywords: winter common wheat, apical meristem, genotype, callus, shoot regeneration.