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Bärbel Röck-Okuyucu, Meltem Bayraktar, Ismail Hakki Akgun, and Aynur Gurel

high labor costs for large-scale production ( Brandle et al., 1998 ; Gantait et al., 2015 ). A genetically homogeneous population, which produces high yields of the desired SGs, can be achieved by in vitro propagation of a selected plant ( Gantait et

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Kourosh Vahdati, Zeinab Maleki Asayesh, Sasan Aliniaeifard, and Charles Leslie

Conventional in vitro propagation can result in production of plants with low survival capacity ( Crane and Hughes, 1990 ). Variable and often insufficient CO 2 concentration, high relative humidity (RH), and accumulation of ethylene or other gases

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W. Msikita, H.T. Wilkinson, and R.M. Skirvin

`Embryonic axes-derived `Burpless Hybrid' cucumber (Cucumis sativus L.) plantlets germinated on Murashige and Skoog (MS) medium supplemented with 16 combinations of BAP and NAA and seedlings derived from whole seeds cultured on semi-solid agar were inoculated in vitro with two isolates (WFU3 and WFM13) of Pythium aphanidermatum. All axes-derived plantlets and whole seedlings inoculated with WFM13 isolates were susceptible to blight and died 2 days after inoculation. Similarly, all seedlings inoculated with WFU3 isolates were killed within 2 days after inoculation; however, the rate of development and severity of blight varied among the axes-derived plantlets. Blight on axes-derived plantlets, regenerated on MS medium supplemented with 2 mg BAP/liter and 0.2 mg NAA/liter, was significantly less than on regenerants cultured on all other amended MS media. On some media, callus developed on crowns and/or primary roots. The presence of callus influenced resistance to Pythium. In a second experiment, axes-derived cucumber regenerants from five genotypes, cultured on MS medium supplemented with 2 mg BAP/liter and 0.2 mg N&A/liter, were compared for their resistance to P. aphanidermatum isolate WFU3. Resistance was significantly greater for `Burpless Hybrid' and `Sweetslice' than for three other genotypes. Chemical names used: 6-benzylaminopurine (BAP); α -naphthaleneacetic acid (NAA).

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Freddi A. Hammerschlag, Ghazala Hashmi, Robin Huettel, Dennis Werner, and David Ritchie

134 WORKSHOP 20 Reevaluating Tissue Culture Techniques for Generating Useful Variation and for in Vitro Screening

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Yung-I Lee, Nean Lee, Edward C. Yeung, and Mei-Chu Chung

This investigation documents the key anatomical features in embryo development of Cypripedium formosanum Hayata, in association with the ability of embryos to germinate in vitro, and examines the effects of culture media and seed pretreatments on seed germination. A better understanding of zygotic embryogenesis for the Cypripedium L. species would provide insights into subsequent germination events and aid in the in vitro propagation of these endangered species. In seeds collected at 60 days after pollination (DAP), soon after fertilization, no germination was recorded. The best overall germination was found at 90 DAP (≈70%), at which time early globular to globular embryos with a single-celled suspensors can be observed. After 135 DAP, the seeds germinated poorly. At this time the inner integument shrinks and forms a tight layer, which encloses the embryo, the so-called “carapace.” Using Nile red stain, a cuticular substance was detected in the carapace, which may play a role in the impermeability of the mature seed and may help the seeds survive in the stringent environment. At maturity (after 210 DAP), the embryo proper has an average size of eight cells along its length and six cells across the width. Lipids and proteins are the main storage products within the embryo. To improve seed germination, experiments were conducted to test the suitability of various media and pretreatments of seeds. When different media were used, except for the Harvais medium at 120 DAP, there was no significant difference in seed germination at three different developmental stages tested. Soaking mature seeds in 1% NaOCl or treating them with ultrasound may slightly increase the germination percentage. For seed germination, our results indicate that the timing of seed collection outweighs the composition of medium and the seed pretreatments.

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Patrick J. Conner

of artificial pollination and pollen storage protocols. Germination tests have generally been considered to be the best in vitro indicator of pollen usefulness ( Galleta, 1983 ). The in vitro germination test assesses the viability of a pollen sample

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José R. Bautista-Aguilar, Lourdes G. Iglesias-Andreu, Jaime Martínez-Castillo, Marco A. Ramírez-Mosqueda, and Matilde M. Ortiz-García

strategies for the conservation of plant germplasm is based on the use of plant tissue culture techniques to help establish an in vitro germplasm bank ( Kendon et al., 2017 ). In vitro conservation in the medium term, allows conserving aseptically, in a small

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Guochen K. Png, Katherine S. Downes, and Beng H. Tan

cuttings to optimize propagation success. Shoot micropropagation, the most common in vitro propagation system, provides an alternative method of propagating plants that is time-consuming or difficult to propagate otherwise ( Offord et al., 2009 ). The

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Dinum Perera and Brian W. Trader

‘Prestige™ Red’ include strong stems, upright growth habit, resistance to stem breakage, and uniform shoot growth, which facilitate floral display and makes it a popular cultivar ( Paul Ecke Ranch, 2008 ). Few studies have reported in vitro organogenesis

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Arancha Arbeloa, Ma Elena Daorden, Elena García, Pilar Andreu, and Juan A. Marín

of ovule culture in breeding programs, primarily for early-maturing peaches and nectarines ( Emershad and Ramming, 1994 ; Pinto et al., 1994 ; Ramming, 1985 ; Ramming et al., 2003 ; Sinclair and Byrne, 2003 ). In addition, in vitro culturing of