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Phu-Long Pham, Ying-Xue Li, He-Rong Guo, Rui-Zhen Zeng, Li Xie, Zhi-Sheng Zhang, Jianjun Chen, Qing-Lian Su, and Qing Xia

orchid seeds produce protocorm during germination, and because in vitro regeneration induces protocorm-like bodies (PLBs) ( Morel, 1960 ; Teixeira da Silva et al., 2015 ), protocorms or PLBs can be easily treated with colchicine for manipulation of

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Jameel M. Al-Khayri, Feng H. Huang, Teddy E. Morelock, and Tahani A. Busharar

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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Xiaoming Wang, Jianjun Chen, Yongxin Li, Qiying Nie, and Junbin Li

‘Jincuilei’ regenerated through indirect shoot organogenesis and its wild-type propagated through cuttings. ( A ) ‘Jincuilei’ with abundant nonopened flowers; ( B ) the wild-type plants with open flowers and flowers to open. In vitro culture has been shown to

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Xiaoling He, Susan C. Miyasaka, Yi Zou, Maureen M.M. Fitch, and Yun J. Zhu

have the potential to enhance disease resistance of crop plants without adversely affecting other important qualities. Genetic transformation involves the insertion of transgenes into totipotent cells that are then regenerated into whole plants. A

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J. M. Al-Khayri, F. H. Huang, and T. E. Morelock

Regenerated spinach (Spinacia oleracea L.) maintained under a 10-h photoperiod (65 uE m-2 s-1) after an incubation period on a GA-containing medium were induced to flower in vitro. The plantlets were regenerated from callus initiated on MS medium with 2.0 mg L-1 kinetin and 0.5 mg L-1 2,4-D and were subsequently transferred to a medium containing 2.0 mg L-1 kinetin, 1.0 mg L-1 GA, and 0.01 mg L-1 2-4,D. While on the regeneration medium, the cultures were exposed to a long-day photoperiod. Regenerants were transferred to an IBA-containing medium for rooting, after which flowering was observed. In vitro flowering plantlets exhibited male and female flowers depending on the sex of the explant donor. Female plantlets developed seeds in the culture vessels. This method of seed production from regenerants can eliminate time-consuming steps in acclimation, transplanting to soil, and plant maintenance.

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Mohsen Hesami and Mohammad Hosein Daneshvar

improvement of genus Ficus via transgenic engineering ( Sharma et al., 2015 ). According to the study that is about shoot regeneration via indirect organogenesis through callus phase conducted by Jaiswal and Narayan (1985) , the maximum 50% regeneration

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J.M. Van Eck and E.D. Earle

Regeneration from VFNT Cherry tomato was optimized prior to transformation. Cotyledon and hypocotyl sections from 7-day-old in vitro grown VFNT Cherry tomato were cultured on medium containing MS salts, B5 vitamins and the following per liter: sucrose, 30g; zeatin, 1 mg; IAA, 0.1 mg; agar, 8.0g or GelriteR, 2.2g. Culture conditions investigated included cotyledon and upper hypocotyl explant lengths, light vs. dark germination, and agar vs. GelriteR. The conditions which resulted in the highest average number of shoots per explant were cotyledon basal explants 2 mm in length, 3.95; cotyledons from dark germination, 6.2; and hypocotyls from light germination, 8.6. An equal number of shoots regenerated on medium containing agar or GelriteR, however, shoots regenerated on medium containing agar were more vigorous. Cotyledon and hypocotyl sections were cocultivated with the Agrobacterium tumefaciens binary vector pBI121 containing the neomycin phosphotransferase II (NPTII) and B-glucuronidasc (GUS) genes. Transformants were selected by regeneration and rooting on medium containing kanamycin. Southern blot and PCR analysis indicated regeneranrs contain the NPTII and GUS genes. Mapping the chromosomal location of the NPTII gene is in progress.

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Whei-Lan Teng, Yann-Jiun Liu, and Tai-Sen Soong

An efficient method for the regeneration of shoots directly from cell suspensions of three commercial cultivars of lettuce (Lactuca sativa L. cv. Great Lakes 659-700, Salad Bowl, and Prize Head) is described. Cell suspensions were prepared by osterizing cotyledon-derived callus for 60 seconds. The effects of callus quality, light intensity, carbohydrate type and concentration, auxins, and cytokinins on cell growth and differentiation in the suspension culture were examined. Among these factors, callus quality and carbohydrates were the most critical. The optimal medium for regeneration of shoots in suspension culture was SH (Schenk and Hilderbrandt) basal medium containing 1000 mg myo -inositol/liter, 1.5% glucose, 0.44 μm BA, and 0.54 μm NAA. The pH of the medium was adjusted to 5.8. Under such condition, hundreds of shoots could be produced from 50 to 55 mg (dry wt) of cell aggregates within 2 weeks. Chemical names used: α-] naphthaleneacetic acid (NAA); indole3-acetic acid (IAA); 6-benzylaminopurine (BA).

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María Victoria González, Manuel Rey, and Roberto Rodríguez

A simple and reliable protocol for plant regeneration from petioles of micropropagated plants of kiwifruit [Actinidia deliciosa (A. Chev) Liang and Ferguson, var. deliciosa `Hayward'] is described. Morphogenic callus was initiated by culturing petioles taken from in vitro-propagated plants. From the media tested, Cheng's K(h) medium plus 0.1 μm IAA, 4.5 μm zeatin, and 2% sucrose was the best for callus induction, maintenance, and shoot bud formation and development. Bases of developed shoots were immersed in 5 mm IBA for 15 seconds; subsequent culture in half-strength K(h) basal medium achieved 82% rooting. Regenerated plantlets were successfully transplanted to soil with 97% survival. Chemical names used: indole-3-acetic acid (IAA); indole-3-butyric acid (IBA); 2-methyl-4-(1H-purin-6-ylamino)-2-buten-1-ol (zeatin).

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J.M. Van Eck and S.L. Kitto

Plant regeneration from callus cultures of mint depended on expiant source, genotype, and culture medium components. Mature embryos, seedling and flower parts, as well as chilled or desiccated immature embryos of peppermint (Mentha piperita L.) and spearmint (Mentha spicata L.) were cultured on a Murashige-Skoog medium containing various combinations of growth factors. Shoots regenerated from callus that developed either on mature peppermint embryos cultured on medium that contained BA at 0.5 mg·liter-1 and NAA at 0.5 mg·liter-1 or on immature peppermint embryos (chilled at SC for 0.6 day or nonchilled) cultured on basal medium containing BA at 1 mg·liter-1 and TIBA at 1 mg·liter-1 Shoots were proliferated, rooted, and acclimated. with 100% survival under greenhouse conditions. Chemical names used: N-(phenylmethyl) -1H-purin-6-amine (BA); 1-naphthaleneacetic acid (NAA); 2,3,5-triiodobenzoic acid (TIBA).