Journal articles: 'Direct shoot regeneration'

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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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Nehra, Narender S., Cecil Stushnoff, and K. K. Kartha. "Direct Shoot Regeneration from Strawberry Leaf Disks." Journal of the American Society for Horticultural Science 114, no. 6 (November 1989): 1014–18. http://dx.doi.org/10.21273/jashs.114.6.1014.

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Abstract An efficient and reproducible method of direct shoot regeneration from leaf disks in strawberry (Fragaria × ananassa cv. Redcoat) has been developed. The influence of hormone concentration, light intensity, orientation of leaf disk, and age of explant source on shoot regeneration was examined. Regeneration of leaf disks reached 94%, with an average of 13 shoots per leaf disk, within 8 weeks when MS salts and B-5 vitamins medium supplemented with 10 μm each of BA and IAA were used. The adventitious shoot meristems initially arose from epidermal or sub-epidermal cells at the periphery of the leaf disks and later from surface cells of the newly regenerated meristems. Shoot regeneration did not depend on light, but low light intensity (12.5 μmol·s−1 m−2) greatly enhanced regeneration. The leaf disks obtained from 30-day-old greenhouse plants and from young runner plants produced shoots at higher frequency than those obtained from 1-year-old plants. Regeneration frequency was higher when the adaxial surface of leaf disks was kept in contact with the medium surface. Shoot regeneration also occurred in nine other genotypes at varying frequencies, but with an intervening short callus phase, except in ‘Veestar’. The technique has potential application for rapid propagation and genetic manipulation of strawberries.

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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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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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Phan, Thi Thu Hien. "Research on the one-step regeneration of sugarcane in KK3 variety (Saccharum officinarum L.) in vitro." Ministry of Science and Technology, Vietnam 64, no. 5 (May 25, 2022): 70–74. http://dx.doi.org/10.31276/vjst.64(5).70-74.

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The research has successfully created the formula for direct regeneration medium of sugarcane variety KK3. In detail, the most suitable medium for direct regeneration from young leaf rolls of sugarcane variety KK3 was RE2 (MS + 5 mg/l α-NAA + 1 mg/l Kinetin), the regeneration rate was 25.27%, the number of shoots formed/1.0 g of young leaf rolls reached 11.56 shoots. The study showed that the position of the young leaf rolls cuttings suitable for direct regeneration was the cuttings located from 2-6 cm away from the growth apex. The shoot regeneration rate reached 25.40%, the regenerating samples were 25.40%, good yield, and a high rate of multiple shoot formation. When using plant cuttings with a thickness of 1 cm, the highest regeneration rate was 25.58%, and the ability to form multiple shoots was high. Pre-cultivation time had a positive effect on the ability to regenerate shoots directly from young leaf coils of sugarcane variety KK3. When conducting pre-cultivation in four days will help increase the regeneration ability of the sugarcane variety, reaching 47.49%. MS medium + 5 mg/l α-NAA + 1 mg/l Kinetin supplemented with 30% coconut water showed the highest regeneration ability, reaching 52.95%, number of shoots reaching 52.33 shoots/1 g of young leaf rolls.

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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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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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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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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 Paeonia mlokosewitschii and P. tenuifolia 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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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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Abeuova, Laura S., Balnur R. Kali, Aizhan O. Rakhimzhanova, Sara S. Bekkuzhina, and Shuga A. Manabayeva. "High frequency direct shoot regeneration from Kazakh commercial potato cultivars." PeerJ 8 (July 13, 2020): e9447. http://dx.doi.org/10.7717/peerj.9447.

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Potato (Solanum tuberosum L.) is the third most economically important crop in the world and has a high nutritional value. In this study, the in vitro culture response of four widely grown in Kazakhstan potato cultivars, Astanalyk, Monument Kunaev, Tokhtar, and Aksor, was investigated using stem and leaf explants. Published protocols were evaluated and optimized to develop a more efficient protocol for the regeneration of plants from local potato cultivars in tissue culture, which is a prerequisite to facilitate potato genome modification. The explants were cultured on solid Murashige and Skoog medium supplemented with different concentrations and combinations of zeatin, 6-benzylaminopurine (BAP), gibberellic acid (GA3), 1-naphthaleneacetic acid (NAA) and indole-3-acetic acid (IAA). The maximum regeneration was induced from the stem internodal explants. A significant effect of the explant source on direct regeneration was confirmed with statistical analysis. The number of shoots obtained from the internode was 10.0 from cv. Aksor followed by cvs. Tokhtar and Astanalyk. The medium DRM-VIII with 1 mg/l zeatin, 0.1 mg/l IAA and 7.0 mg/l GA3 was considered the best for direct shoot regeneration and multiple shoot formation from all cultivars. To conclude, we outline a protocol for direct plant regeneration from four potato cultivars. Our findings suggest commercial cultivars Astanalyk and Aksor are good candidates for developing the genome-edited plants through direct shoot regeneration.

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Jung, Woo-Suk, Ill-Min Chung, Seung-Hyun Kim, Hee-Yeon Chi, Chang Yeon Yu, and Bimal Kumar Ghimire. "Direct Shoot Organogenesis from Lycium chinense Miller Leaf Explants and Assessment of Genetic Stability Using ISSR Markers." Agronomy 11, no. 3 (March 8, 2021): 503. http://dx.doi.org/10.3390/agronomy11030503.

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An efficient in vitro direct shoot regeneration system has been described for Lycium chinense Miller using leaf explants. Influence of various parameters such as growth regulator concentration, explant type, effect of basal salt type, Murashige and Skoog (1962) medium (MS), Schenk and Hildebrandt (1972) medium (SH), Gamborg et al. (1968) medium (B5), and carbon sources (sucrose, maltose, and fructose) on the regenerating shoots has been studied. Micromorphological studies and genetic fidelity of regenerated shoots were assessed and compared with those of the donor plants. Among the different concentrations of plant growth regulator (PGRs) tested, MS supplemented with lower concentration of 6-benzylaminopurine (BAP) (0.5 mgL−1) and thidiazuron (TDZ) (0.5 mgL−1) increased the frequency of shoot. Comparatively, indole-3-butyric acid (IBA) was more effective in the regeneration and growth of the root system. A higher number of root formation (6.67 ± 1.25) was observed when the rooting medium comprised half-strength MS salts supplemented with 3% sucrose. The surviving plantlets were gradually transferred to the greenhouse and natural soil. More than 90% of the plantlets survived and matured within 85 days. Similarity in the band patterns produced by inter simple sequence repeat (ISSR) primers confirmed the genetic stability and uniformity between the regenerated and donor plants. The present optimized direct shoot regeneration system may be useful for mass propagation and improving the genetic traits in L. chinense.

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Mandal, A. K. A., and S. Dutta Gupta. "Direct shoot organogenesis and plant regeneration in safflower." In Vitro Cellular & Developmental Biology - Plant 37, no. 1 (January 2001): 50–54. http://dx.doi.org/10.1007/s11627-001-0010-5.

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Aasim, Muhammad, Nurdan Sahin-Demirbag, Mahmood Khawar, Hayrettin Kendir, and Sebahattin Özcan. "Direct axillary shoot regeneration from the mature seed explant of the hairy vetch (Vicia villosa Roth)." Archives of Biological Sciences 63, no. 3 (2011): 757–62. http://dx.doi.org/10.2298/abs1103757a.

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The hairy vetch (Vicia villosa Roth) is a climbing, prostrate or trailing legume grown as forage.It fixes atmospheric nitrogen, reduces soil erosion and provides an instant mulch. Multiple axillary shoot regeneration from a mature seed explant (zygotic embryo with two cotyledons) was obtained on MS medium containing 0.05 - 1.6 mg/l TDZ with or without 0.10 mg/l IBA. The frequency (%) of shoot regeneration ranged from 45.83-75.00% with a maximum number of 28.6 shoots per explant on MS medium containing 0.20 mg/l TDZ-0.10 mg/l IBA. The mean shoot length decreased proportionately with each increase in TDZ concentration irrespective of the IBA concentration in the culture medium. However, comparing the two types of regeneration media, longer shoots were recorded in the presence of IBA in the culture medium. Regenerated shoots were pulse treated with 50 mg/l IBA for 5, 10 and 20 min for rooting.

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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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Babaei, Nahid, Nur Ashikin Psyquay Abdullah, Ghizan Saleh, and Thohirah Lee Abdullah. "An EfficientIn VitroPlantlet Regeneration from Shoot Tip Cultures ofCurculigo latifolia, a Medicinal Plant." Scientific World Journal 2014 (2014): 1–9. http://dx.doi.org/10.1155/2014/275028.

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A procedure was developed forin vitropropagation ofCurculigo latifoliathrough shoot tip culture. Direct regeneration and indirect scalp induction ofCurculigo latifoliawere obtained from shoot tip grown on MS medium supplemented with different concentrations and combinations of thidiazuron and indole-3-butyric acid. Maximum response for direct regeneration in terms of percentage of explants producing shoot, shoot number, and shoot length was obtained on MS medium supplemented with combination of thidiazuron (0.5 mg L−1) and indole-3-butyric acid (0.25 mg L−1) after both 10 and 14 weeks of cultures. Indole-3-butyric acid in combination with thidiazuron exhibited a synergistic effect on shoot regeneration. The shoot tips were able to induce maximum scalp from basal end of explants on the medium with 2 mg L−1thidiazuron. Cultures showed that shoot number, shoot length, and scalp size increased significantly after 14 weeks of culture. Transferring of the shoots onto the MS medium devoid of growth regulators resulted in the highest percentage of root induction and longer roots, while medium supplemented with 0.25 mg L−1IBA produced more numbers of roots.

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Stamenova-Berberova, Hristina H., and Paul E. Read. "260 Micropropagation of Baptisia bracteata Mnhl. via Direct Regeneration." HortScience 34, no. 3 (June 1999): 487A—487. http://dx.doi.org/10.21273/hortsci.34.3.487a.

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Native plants are often ignored in horticulture because they may lack major ornamental traits and many of them are difficult to propagate. Creamy indigo (Baptisia bracteata Mnhl.) is a North American legume with considerable potential as a container-grown or ornamental plant for managed landscapes. Nodal explants from aseptically germinated seedlings were evaluated for axilary shoot and leaf development. The explants were cultured on Murashige and Skoog medium (MS) containing adenine sulfate at 80 mg•L-1, 30% sucrose, and different levels of N-6-benzyladenine (BA) (0.5,1.0,2.0 mg•L-1) supplemented with indole-3-acetic acid (IAA) (0.05, 0.1 or 0.5 mg•L-1) or with IAA omitted. Shoot regeneration occurred within 2 to 3 weeks. The best medium for shoot regeneration was MS supplemented with BA at 1.0 mg and IAA at 0.1 mg•L-1. Shoots were transferred onto rooting medium consisting of Ω MS supplemented with 1.0 mg alpha-naphthaleneacetic acid (NAA) and 1.0 mg indole-3-butyric acid (IBA)/L and 20% sucrose. Rooting took place within 3 to 5 weeks. Plantlets were then planted in soil mix, placed under a polyethylene tent for 2 weeks, and transferred into the greenhouse for further growth.

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Yousefiara, Mahdieh, Maryam Jafarkhani Kermani, Abdolreza Bagheri, Ali Akbar Habashi, and Hamid Abdollahi. "Induction of Direct Adventitious Shoot Regeneration in Pear (Pyrus communis L.)." Plant Tissue Culture and Biotechnology 24, no. 1 (June 22, 2014): 87–92. http://dx.doi.org/10.3329/ptcb.v24i1.19215.

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Efficient direct shoot regeneration of two pear (Pyrus communis L.), namely 'bartlett' and 'dargazi' was successfully developed for the use in future genetic engineering research. The basal MS and NN media supplemented with different concentrations of TDZ (0, 2.5, 5, 7.5 ?M) or BAP (0, 4, 8, 16 ?M) in combination with NAA (1 ?M) were compared. The result showed that direct adventitious shoot regeneration in pear was highly dependent on genotype, explant type and culture media. 'dargazi' had higher rate of regeneration compared to 'bartlett' and in both cultivars the highest per cent of regeneration was observed in lower sections of the leaves. Although the highest per cent of regeneration in 'bartlett' (38) was attained in the NN medium containing 2.5 µM TDZ and 1 µM NAA, but the differences in shoot regeneration between media containing 5 or 7.5 µM TDZ and 1 µM NAA were not significant. The highest per cent of regeneration in 'dargazi' (56) was obtained in NN medium containing 7.5 µm TDZ and 1 µm NAA. Plant Tissue Cult. & Biotech. 24(1): 87-92, 2014 (June) D. O. I. http://dx.doi.org/10.3329/ptcb.v24i1.19215

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Shen, Xiuli, Vladimir Orbović, Manjul Dutt, William S. Castle, and Frederick G. Gmitter. "Direct Shoot Organogenesis in Murraya paniculata (L.) Jack: A Prerequisite for Genetic Transformation." HortScience 48, no. 7 (July 2013): 938–41. http://dx.doi.org/10.21273/hortsci.48.7.938.

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An efficient in vitro regeneration system through direct shoot organogenesis was established for Murraya paniculata (L.) Jack (Orange Jessamine). Epicotyls, leaves, roots, and cotyledons from in vitro-germinated seedlings and several plant growth regulators (PGRs) were evaluated for their effects on plant regeneration. Longitudinally cut epicotyl segments were observed to be the optimal explants followed by uncut epicotyls (not longitudinally cut). Roots, leaves, and cotyledons were not suitable as explants as a result of little or no shoot induction. Adventitious shoot induction was enhanced by the addition of 6-benzyladenine (BA). The highest percentage of shoot induction (87%) and the greatest number of shoots per explant (12.7) occurred on Murashige and Skoog (MS) medium supplemented with 15 μM BA from longitudinally cut epicotyls followed by 5.2 shoots per explant from uncut epicotyls. Optimal concentration of gibberellic acid (GA3) for shoot elongation was observed to be 15 μM. Eighty-five percent of the regenerated shoots produced roots with an average of three roots per shoot on MS medium supplemented with 5 μM indole-3-butyric acid (IBA). Our protocol for direct shoot organogenesis can potentially lead to the development of a robust method for production of transgenic plants of M. paniculata through Agrobacterium-mediated genetic transformation.

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Colby, Sheila M., Adrian M. Juncosa, and Carole P. Meredith. "Cellular Differences in Agrobacterium Susceptibility and Regenerative Capacity Restrict the Development of Transgenic Grapevines." Journal of the American Society for Horticultural Science 116, no. 2 (March 1991): 356–61. http://dx.doi.org/10.21273/jashs.116.2.356.

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Although grape is easily infected with Agrobacterium and plants can be regenerated routinely, it has proven recalcitrant to the recovery of transgenic plants. Anatomical and histochemical analyses of cocultivated regenerating leaf explants were used to investigate the compatibility of direct shoot organogenesis with Agrobacterium- mediated transformation. Leaves of Vitis vinifera L. cvs. French Colombard and Thompson Seedless were cocultivated with Agrobacterium tumefaciens containing a binary vector carrying kanamycin resistance (APH(3′)II) and (β- glucuronidase (GUS) genes. Explants were cultured on shoot-inducing medium containing levels of kanamycin inhibitory to the formation of untransformed shoots and assayed for GUS expression after 2 or 4 weeks. Cells expressing GUS were most frequently observed either at the cut surface, in vascular bundles, or in inner cortical cells of the petiole, but none of these regions produce adventitious shoots. GUS expression was also frequently found on leaf laminae, where it marked the center of a zone of cross-protected cells, but unwounded lamina cells never participated in shoot regeneration. Cells expressing GUS were found less frequently in the epidermal and subepidermal locations where exogenous, multicellular promeristem initiation occurred. These observations suggest that the direct shoot regeneration system used here could produce chimerally transformed plants, but is unsuitable for the routine production of uniformly transformed plants.

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Mora-Vásquez, Soledad, Silverio García-Lara, Edgardo J. Escalante-Vázquez, and Guy A. Cardineau. "IMPROVEMENT OF DIRECT REGENERATION OF MEXICAN SOYBEAN FROM COTYLEDONARY NODES." Agrociencia 54, no. 3 (December 23, 2020): 387–99. http://dx.doi.org/10.47163/agrociencia.v54i3.1914.

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Plant tissue culture provides an alternative approach to improve the quality of soybean (Glycine max (L.) Merrill) cultivars. This study was undertaken to analyze the susceptibility of Mexican soybean for direct shoot regeneration and to determine the critical factors that affect in vitro performance. Our hypothesis was that Mexican soybean is suitable for in vitro regeneration using a cotyledonary node as explant. The effects of the seed disinfection procedure, soaking pretreatment before germination, soybean variety, as well as the culture medium composition of the shoot induction medium, were evaluated by two split-plot statistical designs. According to the statistical analysis, the seed disinfection procedure, the soaking pretreatment before germination, and the soybean genotype were the factors that brought about a significant effect (p£0.01), while the hormones composition of the shoot induction medium did not have a significant effect. The best response for multiple shoot formation was observed using a chlorine gas seed disinfection method, 3% hydrogen peroxide soaking pretreatment, and Huasteca-100, Nainari and Suaqui soybean genotypes. A robust protocol was developed, and under these selected conditions, it is possible to obtain more than 10 shoots per explant. Well-developed plantlets were obtained after 60 d of in vitro culture.

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Beigi Harchegani, Maryam, Hossein Nazarian, Mahmoud Otroshy, Mohammad Ali Ebrahimi, and Ali Motamedi. "In vitro Regeneration of Alstroemeria cv. ‘Balance’ Based On Direct Organogenesis." Biosciences, Biotechnology Research Asia 14, no. 2 (June 25, 2017): 557–66. http://dx.doi.org/10.13005/bbra/2479.

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ABSTRACT: The present study aimed at improving the efficiency of the in vitro regeneration of Alstroemeria cv. ‘Balance’ protocol through direct organogenesis technique using three different origins of explants (nodal stem, rhizome apical bud, rhizome segments) as two separate factorial experiments with completely randomized design with six replications which were implemented in three stages. Firstly, Direct regeneration, including two factorial experiments to induce regeneration in organogenesis media by utilizing the four NAA concentrations (0, 0.5, 1, 2 mg/l) combined with five BAP concentrations (0, 0.5, 1, 1.5, 2 mg/l) in the first experiment, and TDZ concentrations (0, 0.5, 10 mg/l) in combination with three IBA concentrations (0, 1, 2 mg/l) in the second experiment. Secondly, shoot regeneration and elongation of stems in regenerated buds in the MS basal medium and finally, rooting of the regenerated shoots in the root induction mediums by NAA and IBA. Results of regeneration experiment revealed that, in the culture medium containing 10 mg/l TDZ in combination with 2 mg/l IBA and rhizome apical bud explants, the maximum rates of regeneration percentage, the highest number and length of the shoots were produced. Resulted shoots produced the greatest number of roots and rhizomes in the culture medium comprising 1 mg/l NAA in combination with 2 mg/l IBA as auxins. From the two tested explants, rhizome apical bud, was known as the best explants for shoot regeneration obtained plantlets successfully adapted to environmental conditions and were transferred to the greenhouse. Results of current study indicated that Alstroemeria can be produced through direct organogenesis.

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Meliza, Claude, and Houchang Khatamian. "Direct Regeneration of Pennisetum setaceum `Rubrum'." HortScience 33, no. 3 (June 1998): 462c—462. http://dx.doi.org/10.21273/hortsci.33.3.462c.

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Pennisetum setaceum `Rubrum' (Crimson Fountain Grass) is an attractive ornamental grass. It is adaptable to a wide range of soil types and is drought-tolerant. Nodal explants were taken from containerized plants either grown in greenhouse or outside. The explants were surface-sterilized in 95% ethyl alcohol for 10 min followed by 10 min in 10% Clorox bleach and rinsed three times each for 5 min with sterile double-distilled water. The explants were then cultured in glass tubes of 25 × 150-mm filled with half- and full-strength MS medium supplemented with 1 or 3 mg/L BA and 0.5 mg/L NAA. Shoot regeneration occurred within 1 to 2 weeks. The best medium for shoot regeneration was 1/2MS supplemented with 1 mg/L BA plus 0.5 mg/L NAA. Microshoots were transferred into rooting medium consisting of 1/2MS supplemented with 0.25 NAA. Rooting took place within 5 to 6 weeks. Plantlets were then planted in soilless medium, placed under mist for 1 week, and transferred into the greenhouse for further growth.

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Hesami, Mohsen, and Mohammad Hosein Daneshvar. "In Vitro Adventitious Shoot Regeneration through Direct and Indirect Organogenesis from Seedling-derived Hypocotyl Segments of Ficus religiosa L.: An Important Medicinal Plant." HortScience 53, no. 1 (January 2018): 55–61. http://dx.doi.org/10.21273/hortsci12637-17.

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Ficus religiosa is an important industrial, medicinal, and ornamental plant, so in vitro regeneration is of high paramount in this valuable germplasm. Two efficient protocols were developed for indirect and direct shoot organogenesis through hypocotyl explants. In the first experiment, different concentrations of 2,4-dichlorophenoxyacetic acid (2,4-D) and indole butyric acid (IBA) (0.5, 1.0, and 1.5 mg·L−1) in combination with 6-benzyl amino purine (BAP) (ratio 10:1, respectively) were used for callus formation. Two types of callus were obtained from different concentrations of plant growth regulators (PGRs). Also, 2,4-D produced yellow-brownish and friable callus (Type I), whereas the green and compact callus (Type II) was achieved in IBA. The highest callus fresh weight (2.43 g) was observed in Murashige and Skoog (MS) medium containing 0.5 mg·L−1 2,4-D plus 0.05 mg·L−1 BAP. In the later experiments, various concentrations of thidiazuron (TDZ), 6-furfuryl amino purine (KN), and BAP (0.25, 0.5, 1.0, and 1.5 mg·L−1) in combination with IBA (ratio 10:1, respectively) were applied for shoot regeneration (direct and indirect organogenesis). In shoot regeneration from callus, the highest regeneration frequency (86.66%) and shoot number per callus (4.13) were achieved in MS medium supplemented with 1.5 mg·L−1 BAP plus 0.15 mg·L−1 IBA from type I calli. However, no regeneration was observed in type II calli. In direct shoot regeneration, the highest regeneration frequency (96.66%) and shoot number (6.26) were obtained in the medium mentioned previously. In root induction experiment, different concentrations of naphthalene acetic acid (NAA) and IBA alone or in combination were applied, and MS medium containing 2.0 mg·L−1 IBA along with 0.1 mg·L−1 NAA was the best hormonal balance for root induction. The rooted plantlets’ survival rate was more than 95% in the acclimatization stage. These results demonstrated that the direct regeneration method provides more shoot regeneration frequency and take a less time for shoot organogenesis than the indirect regeneration method. Based on our knowledge, this study is the first report of direct and indirect shoot organogenesis of F. religiosa via hypocotyl from in vitro–grown seedling.

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Yang, Shuyu, Runze Liu, Wenlong Li, Yanan Jing, Solme Pak, and Chenghao Li. "Rapid and Efficient Regeneration of Populus ussuriensis Kom. from Root Explants through Direct De Novo Shoot Organogenesis." Forests 13, no. 5 (May 20, 2022): 806. http://dx.doi.org/10.3390/f13050806.

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Populus ussuriensis is an important tree species with high economic and ecologic values. However, traditional sexual propagation is time-consuming and inefficient, challenging afforestation and wood production using P. ussuriensis, and requires a rapid and efficient regeneration system. The present study established a rapid, efficient, and stable shoot regeneration method from root explants in P. ussuriensis using several plant growth regulators. Most shoot buds (15.2 per explant) were induced at high efficiency under WPM medium supplemented with 221.98 μM 6-BA, 147.61 μM IBA, and 4.54 μM TDZ within two weeks. The shoot buds were further multiplicated and elongated under WPM medium supplemented with 221.98 μM 6-BA, 147.61 μM IBA, and 57.74 μM GA3 for four weeks. The average number and efficiency of elongation of multiplication and elongation for induced shoot buds were 75.2 and 78%, respectively. All the shoots were rooted within a week and none of them showed abnormality in rooting. The time spent for the entire regeneration of this direct shoot organogenesis was seven weeks, much shorter than conventional indirect organogenesis with the callus induction phase, and no abnormal growth was observed. This novel regeneration system will not only promote the massive propagation, but also accelerate the genetic engineering studies for trait improvement of P. ussuriensis species.

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Goswami, Barna, Salim Khan, Tanjina Akthar Banu, Shahina Akter, Mousona Islam, and Ahashan Habib. "In Vitro Mass Propagation of Withania Somnifera (L.) Dunal An Important Medicinal Plant of Bangladesh." Bangladesh Journal of Botany 51, no. 2 (June 28, 2022): 191–97. http://dx.doi.org/10.3329/bjb.v51i2.60414.

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An efficient in vitro regeneration system was developed for Withania somnifera (L.) Dunal through direct and indirect organogenesis from nodal and leaf segment explants. Highest direct shoot regeneration was recorded when nodal explants were cultured on MS with 2.0 mg/l BAP and 0.5 mg/l NAA where the average number of shoots per explants was 10.5. For indirect regeneration remarkable results on shoot initiation were observed when node and leaf segment callus were cultured on MS +1.0 mg/l BAP +0.5 mg/l NAA. It was observed that node callus showed best response for multiple shoot/explants (28.9) on MS +1.0 mg/l BAP +0.5 mg/l NAA. But the mean of shoots from the callus of leaf segment explants was found to be 15.1 on the same media. The best response towards root induction was observed on MS with 2.5 mg/l IBA. The well rooted plants were successfully acclimatized and transferred to soil. Bangladesh J. Bot. 51(2): 191-197, 2022 (June)

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Wang, N., M. J. Liu, Z. H. Zhao, and Z. Y. Qin. "DIRECT SHOOT REGENERATION FROM LEAF EXPLANTS OF SOUR JUJUBE." Acta Horticulturae, no. 840 (August 2009): 287–92. http://dx.doi.org/10.17660/actahortic.2009.840.38.

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Yang, L., C. J. Xu, G. B. Hu, and K. S. Chen. "Direct shoot organogenesis and plant regeneration in Fortunella crassifolia." Biologia plantarum 50, no. 4 (December 1, 2006): 729–32. http://dx.doi.org/10.1007/s10535-006-0117-y.

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Saritha, K. V., and C. V. Naidu. "Direct shoot regeneration from leaf explants of Spilanthes acmella." Biologia plantarum 52, no. 2 (June 1, 2008): 334–38. http://dx.doi.org/10.1007/s10535-008-0068-6.

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Chen, F. C., C. Y. Wang, and J. Y. Fang. "Micropropagation of self-heading Philodendron via direct shoot regeneration." Scientia Horticulturae 141 (June 2012): 23–29. http://dx.doi.org/10.1016/j.scienta.2012.04.011.

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Biswas, Purnendu, Md Harun-Or Rashid, Abul Kashem Chowdhury, and Md Amzad Hossain. "Direct plant regeneration through micropropagation using selected explants of sugarcane." International Journal of Advanced Geosciences 8, no. 2 (November 10, 2020): 244. http://dx.doi.org/10.14419/ijag.v8i2.31100.

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The experiment was conducted at the Laboratory of Biotechnology Division, Bangladesh Sugarcrop Research Institute (BSRI), Ishurdi, Pabna during the period from January 2009 to December 2009 to regenerate plants through micropropagation technique using selected explants of sugarcane. In this experiment shoot tip, leaf segment and leaf roll section explants of three sugarcane varieties (Isd 16, Isd 35 and Isd 36) were cultured on shoot inducing MS media with four combinations of NAA and Kn (NAA2.5Kn0.5, NAA5.0Kn0.5, NAA7.5Kn0.5 and NAA10.0Kn0.5 mg/l) to regenerate plants. The proliferated shoots were multiplied on liquid MS media with five combinations of BAP and Kn (BAP0.25Kn0.25, BAP0.5Kn0.25, BAP0.5Kn0.5, BAP1.0Kn0.5 and BAP1.0Kn1.0 mg/l) and further transferred to root inducing MS media fortified with six different concentrations of NAA (1.0, 2.0, 3.0, 4.0, 5.0 and 6.0 mg/l) for adventitious root formation to raise full-fledged plantlets. The leaf roll section explant of variety Isd 35 cultured on MS medium containing hormonal combination of 7.5 mg/l NAA + 0.5 mg/l Kn produced the highest percentage (83.33%) of shoot regeneration. The number of shoots per explant was also found highest (10.23) in the same explant of same variety with the same hormonal combination. The highest multiplication rate (4.64) was obtained from the liquid MS medium containing hormonal combination of 1.0 mg/l BAP + 0.5 mg/l Kn in the variety Isd 35. The MS medium containing 5.0 mg/l NAA showed better performance for adventitious root induction to raise full-fledged plantlets in all the varieties within nine days of inoculation.

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Thul, Sanjog T., and Arun K. Kukreja. "An Efficient Protocol for High-frequency Direct Multiple Shoot Regeneration from Internodes of Peppermint (Mentha x piperita)." Natural Product Communications 5, no. 12 (December 2010): 1934578X1000501. http://dx.doi.org/10.1177/1934578x1000501223.

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A simple, repeatable and efficient protocol for direct multiple shoot regeneration from internodal explants has been defined in peppermint ( Mentha x piperita var. Indus). In vitro regenerated shoots of peppermint were excised into 4 to 8 mm long internodes and cultured on Murashige and Skoog's medium supplemented with different cytokinins. In the hormonal assay, 3.0 mg L-l zeatin or 6-isopentenyl adenine independently supplemented to half strength MS medium exhibited multiple shoot regeneration, while thiaduzorn (0.1-3.0 mg L−1) showed no morphogenetic effect. A maximum of 85% in vitro cultured explants showed multiple shoot formation with an average of 7 shoots per explant on MS medium supplemented with zeatin. Multiple shoots were initiated within three weeks of cultivation. Internodes with regenerated multiple shoots were transferred to half - strength MS medium without supplementing with any plant growth hormone for shoot elongation and rhizogenesis. Rooted plants acclimatized and grew to maturity under glasshouse conditions. The plantlets developed were phenotypically identical to the parent plant and exhibited 96 % survival.

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Devi, Urmila, and Harinder Singh Rattanpal. "In vitro Plant Regeneration in Cleopatra (Citrus reshni Hort. ex Tan.) by Direct Organogenesis." Plant Tissue Culture and Biotechnology 28, no. 2 (December 5, 2018): 251–60. http://dx.doi.org/10.3329/ptcb.v28i2.39683.

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Maximum shoot regeneration (87.85%) and number of shoots per explant in the epicotyl segments having longitudinal cut in Citrus reshni var. Cleopatra were obtained on MT medium supplemented with BAP (2 mg/l) + NAA (0.2 mg/l). Maximum shoot length (5.10 cm), average leaf number per shoot (12), leaf length (2.12 cm) and width (1.5 cm) in Cleopatra were obtained on MT medium supplemented with BAP (2 mg/l) + NAA (0.2 mg/l). The maximum in vitro rooting (78.88 %) in the excised shoots occurred in liquid medium on MS supplemented with BAP (0.5 mg/l) + NAA (0.5 mg/l). Plant Tissue Cult. & Biotech. 28(2): 251-260, 2018 (December)

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Uysal, Hüseyin. "In vitro propagation of black cumin (Nıgella satıva L.) plants." Genetika 53, no. 1 (2021): 295–303. http://dx.doi.org/10.2298/gensr2101295u.

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This study was carried out to determine in vitro development using Black cumin leaf and stem explants. ?ameli black cumin variety was used as plant material. Five different nutrient mediums (1. LS2.5, 2. MS, 3. MS + 0.5 mg.l-1 IAA, 4. MS + 0.5 mg.l-1 BAP, 5. MS + 0.5 mg.l-1 IAA + 0.5 mg.l-1 BAP) containing 30 g sugar were used in this study. As a result of the research, 100% callus formation was detected in the stem explants cultured in the number 1 and number 5 mediums. These were followed by stem explants cultured in medium 4 with a success rate of 96%. Of this rate, 66% was shoot formation, and 30% was callus formation. Direct shoot regeneration was performed only on stem explants cultured in mediums 4 and 3, with a 66% success rate in medium four and a 36% success rate in medium 3. The highest plant regenerations from calluses were gained from stem explants (273.3%) in medium 4, followed by calluses gained from leaf explants (262.5%) in the same medium. These were followed by cultures in medium 3, with calluses derived from stem explants (255%) and leaf explants (150%). No plant regeneration was determined from calluses gained in the medium 1. Thus it is evident that high auxin content and auxin-cytokinin balanced mediums encouraged callus formation in the black cumin plants. The addition of only IAA or BAP to the medium promoted shoot formation in the stem explants, but direct shoot regeneration was not thereby achieved from the leaf explants. These results show that, for in vitro clonal propagation studies done on black cumin plants, a high auxin containing medium is preferable if the aim is callus formation. If the aim is direct shoot regeneration, BAP or other cytokinin-containing medium is preferred.

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Nadolska-Orczyk, Anna, Lidia Miłkowska, and Wacław Orczyk. "Two ways of plant regeneration from immature cotyledons of pea." Acta Societatis Botanicorum Poloniae 63, no. 2 (2014): 153–57. http://dx.doi.org/10.5586/asbp.1994.020.

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Two different systems of plant regeneration via organogenesis and embryogenesis from immature cotyledons of pea (Pisum sativum L.) were developed. The first system was direct multiple shoot regeneration from the proximal to the embryo axis, injured part of cotyledon. The ability of six Polish cultivars to shoot formation was very high. The percent of regenerating cotyledons was from 73 to 86 and mean number of shoots from 3.5 to 9.9 after seven weeks of culture. This multiplification could be prolonged for next several months. The second system was somatic embryogenesis, initiating from the same part of cotyledon simultaneously with slowly proliferating callus. Only three out of six cultivars formed embryoids. The differences of ability to embryo formation ranged between 43% of responding explants for Heiga cultivar to only 6% for Cud Ameryki. The mean number of embryoids was from 4.2 to 2.3.

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Balilashaki, Khosro, Maryam Vahedi, and Roghayeh Karimi. "In vitro Direct Regeneration from Node and Leaf Explants of Phalaenopsis cv. Surabaya." Plant Tissue Culture and Biotechnology 25, no. 2 (January 4, 2016): 193–205. http://dx.doi.org/10.3329/ptcb.v25i2.26254.

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An efficient and reproducible procedure for the direct regeneration of phalaenopsis cv. Surabaya using of nodal explants and leaf segments derived from in vitro flower stalk was conducted. Three experiments were carried out for shoot development and subsequent plant regeneration: Direct shoot regeneration from nodal explants of Phalaenopsis cv. Surabaya flower stalks on MS added with different combination of NAA and BAP, direct regeneration of protocormlike bodies (PLBs) from leaf explants in a MS with different concentrations of the TDZ, acclimatization of regenerated plantlets in different mixture of components and nutrients. The results showed that 5 mg/l BAP and 2 mg/l NAA were most effective concentration for shoot regeneration, regenerated shoots were cultured on half strength of MS containing activated charcoal, IAA and NAA at various concentrations, highest number of root (6.7) was obtained in higher concentration of NAA (2 mg/l). TDZ induced a higher frequency of embryogenesis from leaf explants than BAP, the highest number of embryos per explant was 22.45 at 3 mg/l TDZ. Altogether, BAP at higher concentration (10 mg/l) with 1 mg/l NAA had the highest enhancement on the amount of direct embryogenesis. In our investigation 87.20% plantlets via nodal explants survived acclimatization process in medium containing cocopeat and coal (1 : 1). The survival rate of regenerated plantlets via nodal explants (82.07%) was more than of regenerated plantlets via leaf explants (70.47). This protocol provides the basis for further investigation on micropropagation and breeding programs in Phalaenopsis cv. Surabaya.Plant Tissue Cult. & Biotech. 25(2): 193-205, 2015 (December)

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Dhital, Shambhu P., Hak T. Lim, and Hira K. Manandhar. "Direct and Efficient Plant Regeneration from Different Explants Sources of Potato Cultivars as Influenced by Plant Growth Regulators." Nepal Journal of Science and Technology 12 (July 21, 2012): 1–6. http://dx.doi.org/10.3126/njst.v12i0.6471.

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Response of widely grown potato cv. Superior and newly developed cvs. Gui valley and Bora valley to plant growth regulators (PGRs) for direct plant regeneration from internode, leaf blade and petiole explants were investigated. The explants were cultured on a MS solid medium supplemented with different concentrations and combinations of 6-benzylaminopurine (BAP), 1-naphthaleneacetic acid (NAA), zeatin, indole-3-acetic acid (IAA) and gibberellic acid (GA3). Potato cv. Superior, regenerated direct shoot without callus and root formation on MS solid medium supplemented with BAP or zeatin, proliferous roots were produced on NAA or IAA supplemented medium and only some calli were produced on GA3 supplemented medium. The regeneration response varied with different concentrations of PGRs, singly and also in combinations. In the case of combined application of PGRs, the highest shoot regeneration (75.3%) and number of shoot per explant (11.5) and number of roots per explant (7.0) were obtained from the MS solid medium supplemented with zeatin (2 mg l-1), NAA (0.1 mg l-1) and GA3 0.1 mg l-1). Among the three types of explants evaluated, internodes produced the highest number of shoots and roots for both potato cvs. Gui valley and Bora valley, and petiole produced the least number of shoots and roots. The regenerated shoots were rooted in PGRs-free MS solid medium and successfully established under glasshouse condition. Leaf, flower, and tuber morphology were identical to in vitro control and mother plants in the same conditions. This optimized regeneration system can be used for rapid shoot proliferation and also for gene transformation.DOI: http://dx.doi.org/10.3126/njst.v12i0.6471 Nepal Journal of Science and Technology 12 (2011) 1-6

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Heidarpoor, Mansoor, Mansoor Kalantar, Mahmoud Khosroshaheli, and Eslam Majidi Hervan. "Investigation on the direct shoot regeneration in date palm CV.Pyarum and possible genetics changes in induced shoots." Nexo Revista Científica 33, no. 02 (December 31, 2020): 423–30. http://dx.doi.org/10.5377/nexo.v33i02.10781.

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The palm (Phoenix Dactylifera) is one of important trees, and is economically important in south of Iran. Date palm is propagated by the offshoots, number of which is limited. Therefore, adult Date palms produce shoot tips and axillary shoot meristems through the use of a tissue culture. This study was conducted to perform in -vitro tissue culture direct shoot regeneration and determine the best combination of plant regulators and other conditions. To achieve organogenesis and multiplication directly from shoot tips and axillary shoot meristems of Date palm (Phoenix Dactylifera L. Var Pyarum) was used without callus formation. Direct regeneration of vegetative buds minimizes the risk of somaclonal variation among plant regenerates. Results revealed that MS medium supplemented with 4mg/l of kinetin and 3 mg/l of IAA or 2mg/l of BA and 4 mg/l of NAA was the best formation from shoot tip after 16-20 weeks. Subculture per month was evaluated at following conditions: temperature for growth of 27±1°C during the lighted period and 22±1°C during the dark period.

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Arrillaga, I., and S. A. Merkle. "Regenerating Plants from in Vitro Culture of Black Locust Cotyledon and Leaf Explants." HortScience 28, no. 9 (September 1993): 942–45. http://dx.doi.org/10.21273/hortsci.28.9.942.

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A protocol to achieve efficient plant regeneration from juvenile black locust (Robinia pseudoacacia L.) explants is described. Direct adventitious shoots were induced from cotyledon explants on woody plant medium containing 22.2 μm BA and 0.4 μm 2,4-D. Shoots developed and new shoots were induced when the explants were transferred to medium without growth regulators. The effect of dark incubation on shoot regeneration from cotyledons indicated that 15 days of darkness resulted in a high regeneration frequency (91.7%). Adventitious shoot formation also was induced from sections of in vitro-derived leaves cultured in darkness on Murashige and Skoog medium supplemented with 4.4 μm BA and 24.6 μm IBA. A shoot regeneration frequency of 89% was obtained when explants were subcultured on a medium containing 4.4 μm BA and 0.5 μm IAA. Shoots were rooted on Schenk and Hildebrandt medium with or without IBA. Plantlets were acclimatized and grown in the greenhouse. Chemical names used: N -(phenylmethyl)-1H -purin-6-amine (BA); 2,4-dichlorophenoxyacetic acid (2,4-D); indole-3-acetic acid (IAA); indole-3-butyric acid (IBA).

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Beniwal, V., Sarina, S. Narkhede, J. Laura, and Puneet Beniwal. "In Vitro Studies on Direct Regeneration of Nothapodytes Foetida using Shoot Tip and Nodal Explants." Journal of Non-Timber Forest Products 19, no. 3 (September 1, 2012): 191–94. http://dx.doi.org/10.54207/bsmps2000-2012-349n16.

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The effect of different concentrations of auxins and cytokinins either alone or in combination with additives was assessed to be useful on the direct regeneration of shoot tip and nodal explants of N.foetida. BA and IBA proved comparatively better than others. The highest direct regeneration percent (95.0, 92.0 in shoot tip and nodal explants respectively) was observed on the MS medium supplemented with BA (3.0mgL-1) + IBA (1.0mgL-1) and additives (adenine sulphate (50mgL-1) + glutamine (100mgL-1) + L-arginine (25mgL-1) + citric acid (0.0025%) + ascorbic acid (0.005%). Shoot tips as explants performed better by giving higher regeneration percentage and better growth of propagules in comparison to nodal explants in the establishment media.

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Saha, P., M. Afrin, AKM Mohiuddin, and AM Shohael. "In vitro Regeneration of Grass Pea (Lathyrus sativus L.)." Jahangirnagar University Journal of Biological Sciences 4, no. 2 (May 18, 2016): 1–8. http://dx.doi.org/10.3329/jujbs.v4i2.27790.

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In vitro regeneration protocol for grass pea (Lathyrus sativus L.) was optimized using different concentrations and combinations of growth regulators. Direct shoot regeneration obtained through shoot organogenesis from different explants of grass pea cultured on MS medium supplemented with Gamborg B5 vitamin containing 6-benzylaminopurine (BAP), Thidiazuron (TDZ) and ?-naphthalene acetic acid (NAA). Highest percentage of shoots were obtained at 4.0 mg/l of BAP on nodal explants. Stunted multiple shoots were developed from nodal explants while 1.5 mg/l TDZ was used. About 56% of direct shoots were also obtained, while the combination of BAP (4.0 mg/l) and NAA (0.5 mg/l) were used. Regenerated plantlets were rooted most effectively (40%) in rooting medium containing half strength of MS basal medium containing 1.0 mg/l NAA. Well rooted plantlets were further successfully acclimatized to ambient humidity level and grown in controlled environment until hardening.Jahangirnagar University J. Biol. Sci. 4(2): 1-8, 2015 (December)

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Li, Qiansheng, Min Deng, Jie Zhang, Wei Zhao, Yigang Song, Quanjian Li, and Qingjun Huang. "Shoot Organogenesis and Plant Regeneration from Leaf Explants ofLysionotus serratusD. Don." Scientific World Journal 2013 (2013): 1–7. http://dx.doi.org/10.1155/2013/280384.

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The gesneriaceous perennial plant,Lysionotus serratus, has been used in traditional Chinese medicine. It also has a great development potential as an ornamental plant with its attractive foliage and beautiful flowers. An efficient propagation and regeneration system via direct shoot organogenesis from leaf explant was established in this study. High active cytokinin (6-benzyladenine (BA) or thidiazuron (TDZ)) was effective for direct organogenesis of initial induction. Murashige and Skoog (MS) growth media containing 0.5 mg L−1BA alone or with combination of 0.1 mg L−1 α-Naphthaleneacetic acid (NAA) were the most effective for shoot proliferation. High BA concentration (1.0 mg L−1) in the media caused high percentage of vitrified shoots though they introduced high shoot proliferation rate. Histological observation indicated that adventitious shoot regeneration on the medium containing 0.5 mg L−1BA alone occurred directly from leaf epidermal cells without callus formation. Regenerated shoots rooted well on medium containing half-strength MS medium with 0.5 mg L−1indole-3-butyric acid (IBA) and indole-3-acetic acid (IAA), and the plantlets successfully acclimatized and grew vigorously in the greenhouse with a 94.2% and 92.1% survival rate.

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Nada, Shadia, Siva Chennareddy, Stephen Goldman, Sairam Rudrabhatla, Shobha Devi Potlakayala, Puthyaparambil Josekutty, and Karelia Deepkamal. "Direct Shoot Bud Differentiation and Plantlet Regeneration from Leaf and Petiole Explants of Begonia tuberhybrida." HortScience 46, no. 5 (May 2011): 759–64. http://dx.doi.org/10.21273/hortsci.46.5.759.

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We report here an efficient and high-frequency protocol for direct shoot bud differentiation from mature leaves and petioles of greenhouse-grown Begonia tuberhybrida plants. Shoot buds were induced directly on the adaxial surface of leaf tissues from not only at the cut ends, but also across the entire surface of both leaf and petiole segments. The highest frequency of shoot bud formation was 90%, and the maximum number of shoots (132) per leaf explant was achieved with modified Murashige and Skoog (MS) (Murashige and Skoog, 1962) media supplemented with 1.0 mg·L−1 α-naphthalene acetic acid (NAA) and 2.0 mg·L−1 thidiazuron (TDZ). In petioles, the highest frequency of shoot buds was 82%. A maximum number of 33 shoots per explant was achieved with 0.5 mg·L−1 NAA and 2.0 mg·L−1 TDZ. The number of shoots produced in both explants was drastically reduced in the treatment with benzyl-aminopurine (BAP) alone or in combination with NAA and/or TDZ. The regenerated shoots were rooted on MS medium supplemented with 0.5 mg·L−1 NAA. All the elongated shoots developed into complete, rooted plantlets within 3 months. All the plantlets were successfully transferred to soil in pots in the greenhouse and they produced morphologically normal flowers. Chemical names used: α-naphthalene acetic acid (NAA), N-phenyl-N′-1,2,3-thiadiazol-5ylurea (TDZ; thidiazuron), 6-benzyl aminopurine (BAP)

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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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Kumar, Suresh, and Amaresh Chandra. "Direct Plant Regeneration via Multiple Shoot Induction in Stylosanthes seabrana." CYTOLOGIA 74, no. 4 (2009): 391–99. http://dx.doi.org/10.1508/cytologia.74.391.

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Pati, Pratap Kumar, Madhu Sharma, Anil Sood, and P. S. Ahuja. "Direct shoot regeneration from leaf explants of Rosa damascena Mill." In Vitro Cellular & Developmental Biology - Plant 40, no. 2 (March 2004): 192–95. http://dx.doi.org/10.1079/ivp2003503.

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Sivanesan, I., and Byoung Ryong Jeong. "Direct shoot regeneration from nodal explants of Sida cordifolia Linn." In Vitro Cellular & Developmental Biology - Plant 43, no. 5 (October 9, 2007): 436–41. http://dx.doi.org/10.1007/s11627-007-9090-1.

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Ghimire, B. K., E. S. Seong, E. J. Goh, N. Y. Kim, W. H. Kang, E. H. Kim, C. Y. Yu, and I. M. Chung. "High-frequency direct shoot regeneration from Drymaria cordata Willd. leaves." Plant Cell, Tissue and Organ Culture (PCTOC) 100, no. 2 (November 3, 2009): 209–17. http://dx.doi.org/10.1007/s11240-009-9627-6.

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Haddadi, Fatemeh, Maheran Abd Aziz, Hossein Kamaladini, and Seyed Ali Ravanfar. "Thidiazuron- and Zeatin-induced High-frequency Shoot Regeneration from Leaf and Shoot-tip Explants of Strawberry." HortTechnology 23, no. 3 (June 2013): 276–81. http://dx.doi.org/10.21273/horttech.23.3.276.

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Strawberry (Fragaria ×ananassa cv. Camarosa) was evaluated to determine a high-frequency shoot regeneration response for leaf and shoot-tip explants. For direct organogenesis from strawberry leaves, combinations of moderate concentrations of thidiazuron [TDZ (0, 2, and 4 μm)] and 6-benzylaminopurine [BAP (0, 4, and 9 μm)] added into medium containing Murashige and Skoog (MS) basal salts were compared. The most shoots regenerated per leaf explant were observed with 4-μm TDZ. Regeneration from shoot tips was evaluated with 0-, 2-, 4-, 8-, and 16-μm zeatin, kinetin, or 6-α,α-dimethylallylamino purine (2ip) tested individually. Optimum shoot proliferation was achieved from shoot-tip explants on medium containing 4-μm zeatin. Rooting was best without cytokinins in the medium; however, adequate rooting was obtained on the 4-μm zeatin treatment as well.

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Gill, R., and Praveen K. Saxena. "Direct somatic embryogenesis and regeneration of plants from seedling explants of peanut (Arachis hypogaea): promotive role of thidiazuron." Canadian Journal of Botany 70, no. 6 (June 1, 1992): 1186–92. http://dx.doi.org/10.1139/b92-147.

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An efficient procedure has been developed for inducing direct somatic embryogenesis, organogenesis, and regeneration of plants from tissue cultures of peanut (Arachis hypogaea L.). Thin transverse sections of the cotyledons and juvenile leaves were cultured on Murashige and Skoog medium supplemented with N6-benzylaminopurine (BAP) or a substituted phenylurea, thidiazuron (TDZ). Somatic embryos or shoot buds differentiated from cut surfaces of the cotyledons and midrib region of the leaves. The application of BAP induced differentiation of shoot buds whereas the treatment with TDZ resulted in the production of somatic embryos. Somatic embryos developed into plants after subculturing on a basal meduim. Agar-solidified medium was found to be superior to the liquid medium for the development of embryos and shoot buds. The procedure of TDZ-induced somatic embryogenesis and plant regeneration was successfully applied to three genotypes of peanut. A distinct feature of this study is the induction of the morphogenic competence in cultures of seedling expiants of peanut that so far have remained recalcitrant to somatic embryogenesis in vitro. Key words: peanut, Arachis hypogaea, shoot regeneration, somatic embryogenesis, thidiazuron, plant regeneration.

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Journal articles on the topic 'Direct shoot regeneration'

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