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Kimberly A. Pickens, James M. Affolter, Hazel Y. Wetzstein, and Jan H.D. Wolf

Tillandsia eizii is an epiphytic bromeliad that due to over-collection, habitat destruction, and physiological constraints has declined to near threatened status. This species exhibits high mortality in the wild, and seed are characterized by low percentages of germination. As a means to conserve this species, in vitro culture protocols were developed to enhance seed germination and seedling growth. A sterilization protocol using 70% ethanol for 2 minutes followed by 2.6% NaOCl for 40 minutes disinfested seed and promoted seedling growth. Sucrose incorporated into the culture medium had no effect on germination or growth, while NAA inhibited growth, but not germination. Cultures maintained under a 16-hour photoperiod at 22 °C exhibited greater growth than those grown at 30 °C. Seed that germinated in the dark remained etiolated and failed to develop even after transfer to light conditions. Plants grown in vitro were successfully acclimatized and transferred to the greenhouse. Over 86% survival and rapid growth were obtained with either an all-pine-bark medium, or a mixture of 2 redwood bark: 2 fir bark: 2 potting mix: 1 perlite. This demonstrated that in vitro culture of seed may be used to rapidly produce large numbers of T. eizii, and thus can be used for the conservation and reintroduction of this species.

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Natalia R. Dolce, Luis A. Mroginski, and Hebe Y. Rey

Sansberro et al. (2000) who have found that low-temperature treatment increased the germination percentage of I. paraguariensis isolated embryos and cut pyrenes cultured in vitro. In many species, exposition of seeds to low temperatures decreases the

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Song-jun Zeng, Zhi-lin Chen, Kun-lin Wu, Jian-xia Zhang, Cheng-ke Bai, Jaime A. Teixeira da Silva, and Jun Duan

. Asymbiotic seed germination and in vitro seedling development of Nothodoritis zhejiangensis . ( A ) Flowering specimen of N. zhejiangens growing on the tree branches of Cornus officinalis . ( B ) Seed germination and protocorm development in vitro. ( C

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A. Raymond Miller, Joseph C. Scheereus, Patricia S. Erb, and Craig K. Chandler

1 To whom reprint requests should be addressed. Journal Article no. 249-90. Salaries and research support provided in part by state and federal funds appropriated to the Ohio Agricultural Research and Development Center and a Seed Grant to A.R. M

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M.A.R. Mian, R.M. Skirvin, M.A. Norton, and A.G. Otterbacher

., Baton Rouge, LA 70803. 4 Principal Research Specialist in Horticulture. Funds for this research were provided in part by the Illinois Agricultural Experiment Station. Special thanks to Gene Galletta and Adam Dale for their encouragement in this

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Ernie DeMarie, Molly Weimer, and K.W. Mudge

Green pods of the C. reginae orchid were collected from a bog near Ithaca, NY. Pods were surface sterilized, and seeds were plated on agar media. The 8 germination treatments were arrange in a complete 3 way factorial consisting of 2 basal media (fish emulsion, FE vs. yeast extract, YE), 2 media pHs (4.8, 6.8), and 2 temperature regimes (constant 24 C vs. 6 wks. at 5 C prior to transfer to 24 C). Sequential stages of development included embryo enlargement and rupture of the testa (l), root elongation (2), leaf primordium development (3) and finally rhizoid development with or without protocorm greening (4). After 4 months post sowing, germination (stage 1 or beyond) was 56% and 0% on FE at pH 4.8 and 6.8 respectively, and 45% and 78% on YE at pH 4.8 and 6.8 respectively. Protocorm development from unchilled seeds after 4 months was greatest on YE at the lower pH, with 14% reaching stage 3 or 4, as contrasted to only 5% reaching stage 2 (none beyond), for the other germinated treatments. Chilled seeds had higher germination for all treatments but no development beyond stage 1 at 4 months post sowing.

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James E. Henrich, Dennis P. Stimart, and Peter D. Ascher


Seeds of 29 terrestrial orchid species representing 15 genera were surface sterilized by immersion in 0.5% sodium hypochlorite containing a wetting agent, washed, sown on a completely defined, semisolid embryo culture medium containing macro- and microelements, sucrose, amino acids, and vitamins, and incubated in the dark at 25°C. Six months after sowing, 16 species from 9 genera germinated and continued development while 13 species from 10 genera failed to germinate. Species of Cypripedium, Goodyera, Platanthera and Spiranthes differed in response in that one or more of each germinated and one or more did not. Seedling development was similar for most germinating species and progressed to the formation of a shoot or shoot initial in all but one. Apparently the mycorrhizal association thought to be required for terrestrial orchid seed germination and early seedling development can be replaced with aseptic culture on a completely defined medium for many terrestrial orchids.

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Stephen Patrick Greer and Timothy A. Rinehart

. paniculata and there are no published reports as to the relative effectiveness of this mutagen in these species. In addition, nothing is known about the effects of EMS on the viability, dormancy, or germination of Hydrangea seeds. We performed a series of

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Takahiro Tezuka, Hisa Yokoyama, Hideyuki Tanaka, Shuji Shiozaki, and Masayuki Oda

. latifolia and I. rotunda were sown on wet filter paper, germination was not observed in either the 2006–07 or 2007 seasons. In contrast, when cut seeds or embryos were cultured in vitro, quarter-seeds without endocarp (sown on 20 July 2007) and embryos of

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Xiuli Shen and Myeong-Je Cho

factors governing dormancy and germination in sugar pine. Ultimately, the objective of this study was to develop a reliable and rapid in vitro germination protocol in sugar pine. To achieve this goal, we sought to 1) examine seed morphology and structure