春石斛為具商業價值的觀賞植物，其育種因雜交不親和性而面臨雜交障礙。為探討春石斛之雜交授粉後發育情形及雜交障礙類型，本篇研究以Den. Comet King ‘Akatsuki’為主要材料，與其他13種春石斛進行雜交試驗。實驗說明春石斛授粉後雌配子體才開始發育生長，授粉後1週花粉已萌芽，授粉後2週花粉管已進入子房，授粉後3週花粉管隨著子房膨大更加深入，授粉後12週胎座上有數以千計的小突起，授粉後20週已生成眾多具胚種子，授粉後6個月種子成熟。本研究之春石斛雜交障礙可分為授粉後1-2週落花及生成無胚種子兩種，前者為受精前親本間雜交不親和或花粉不正常造成，後者則為雜種胚敗育死亡或未受精。深入探討產生雜交障礙的2種春石斛，Den. Yellow Song ‘Candy’花粉46.0%為正常的四分體，推測雜交障礙為親本間不親和所致；Den. Love Memory ‘Fizz’花粉僅14.8%為正常的四分體，配子體減數分裂不正常，推測其為異源三倍體。關於雜交障礙之克服，本實驗使用切除花柱授粉、注入花柱分泌物及酒精處理等3種方法，其中注入花柱分泌物之效果最佳，花粉管生長速度快，且授粉後12週觀察種子量有增加趨勢。

Nobile-type Dendrobium is an attractive ornamental plant of great business value. One crucial barrier, however, underlies its breeding: the cross-incompatibilities among Dendrobium cultivars. To investigate the development of cross-pollinated Dendrobium, as well as to identify the crossing barriers, interspecific crosses between Den. Comet King ‘Akatsuki’ and 13 other Dendrobium cultivars were conducted. The results showed that female gametophyte began to develop after pollination. One week after pollination, the pollen germinated; in the second week, the pollen entered the ovule. In three weeks, the ovule enlarged and the pollen tubes grew deeper. After 12 weeks, thousands of protrusions were observed on surface of the placenta. By the end of the twentieth week, seeds with functional embryos were formed. Six months after pollination, the seeds were fully matured. Based on the observations, crossing barriers can be coded into two major types: flowers dropping 1-2 weeks after pollination and seeds forming without embryo. The former is caused by either cross-incompatibility or abnormal pollen grain; the latter is caused by either embryo abortion or unfertilized ovule. In the current study, two of the 13 target Dendrobium flowers confronted conspicuous crossing barriers: the Den. Yellow Song ‘Candy’ and the Den. Love Memory ‘Fizz.’ Sporad type observations manifested that Den. Yellow Song ‘Candy’ yielded 46.0% of normal tetrads. It was considered that the crossing barrier is a result of cross-incompatibility. As for Den. Love Memory ‘Fizz,’ merely 14.8% of the pollen were found normal tetrads, which is a result of the gametophyte undergoing aberrant meiosis; on reasonable inference is that Den. Love Memory ‘Fizz’ is an allotriploid. In reducing the above-mentioned crossing barriers, three treatments were employed: cut-column pollination, stigmatic exudate injection and ethanol treatment. Among all, stigmatic exudate injection exerted the most significant effect. After the treatment, pollen tubes grewing faster, and the amount of seeds was increased, as was observed by the end of the twelfth week.

山本洋蘭園。2021年6月16日，取自: https://yamamotodendrobiums. blogspot.com/
尤彤蒂（2013）。春石斛種子貯藏、發芽及微體繁殖與仙履蘭芽體增殖之研究。國立屏東科技大學熱帶農業暨國際合作系博士論文，屏東縣。2021年6月15日，取自「全國博碩士論文資訊網」：https://hdl.handle.net/11296/v4qeqx
金石文、李鎮宇、陳福旗（2020）。台灣春石斛育種的策略。臺灣蘭花之育種（頁106-112）。
王雁、李振堅、彭紅名（2007）。石斛蘭─資源．生產．應用。中國林業出版社。
邱金春、王三太（2011）。遠緣雜交育種障礙及克服策略。農業試驗所技術服務，22，19-22。
金石文、呂廷森、陳福旗（2012）。春石斛的品種趨勢及新品種開發。2011年花卉研究團隊成果發表會專刊，97-106。
胡適宜（1990）。被子植物胚胎學。曉園出版社，307頁。
張焜標、許秋容、張雯婷（2010）。珊瑚樹小孢子的發育. 林業研究季刊， 32，51-61。
許圳塗（1993）。果樹異交授粉及自交不親和性之特性。果樹育種研習會專刊，37，19-32。
許嘉錦（2020）。符合商業需求之春石斛蘭育種及成果。臺中區農業改良場特刊，142，96。
蔡奇助、莊畫婷（2009）。遠緣雜交與分子遺傳鑑定技術在蝴蝶蘭育種之應用潛力。農業生技產業季刊，17，37-45。
魏芳明（2011）。春石斛蘭研究現況與展望。農業試驗所特刊，154，63-70。
魏芳明（2013）。蘭花雜交育種障礙之探討。台中區農業改良場101年專題討論專集特刊，116，307-311。
Allen, A., and S. Hiscock. (2008). Evolution and phylogeny of self-incompatibility systems in angiosperms. In self-incompatibility in flowering plants. Springer, Berlin, Heidelberg. pp.73-101
Ascher, P. D., and L. W. Drewlow. (1971). Unilateral interspecific incompatibility in Lilium. Yearbook of the North American Lily Society 24: 70-74.
Bolaños-Villegas, P., S. W. Chin, and F. C. Chen. (2008). Meiotic chromosome behavior and capsule setting in Doritaenopsis hybrids. J. Am. Soc. Hortic. Sci. 133: 107-116.
Chase, M. W., K. M. Cameron, J. V. Freudenstein, A. M. Pridgeon, G. Salazar, C. Van den Berg, and A. Schuiteman. (2015). An updated classification of Orchidaceae. Bot. J. Linn. Soc. 177: 151-174.
Chuang, H. T., K. L. Huang, and S. T. Hsu. (2018). Cut-column pollination method to overcome pollination barrier in Phalaenopsis. In Orchid Propagation: From Laboratories to Greenhouses—Methods and Protocols. Humana Press, New York. pp. 241-256.
De Nettancourt, D. (1977). Incompatibility in angiosperms. Springer, Berlin, Heidelberg. 230 pp.
Gahan, P. B. (1984). Plant histochemistry and cytochemistry. Academic Press, London. 301 pp.
Hancock, C. N., K. Kondo, B. Beecher, and B. McClure. (2003). The S-locus and unilateral incompatibility. Philos. Trans. R. Soc. Lond., B, Biol. Sci. 358: 1133-1140.
Hiscock, S. J., and H. G. Dickinson. (1993). Unilateral incompatibility within the Brassicaceae: further evidence for the involvement of the self-incompatibility (S)-locus. Theor. Appl. Genet. 86: 744-753.
Hiscock, S. J., and A. M. Allen. (2008). Diverse cell signalling pathways regulate pollen-stigma interactions: the search for consensus. New Phytol. 179: 286-317.
Iwano, M., and S. Takayama. (2012). Self/non-self discrimination in angiosperm self-incompatibility. Curr. Opin. Plant Biol. 15: 78-83.
Kamemoto, H., T. D. Amore, and A. R. Kuehnle. (1999). Breeding Dendrobium Orchids in Hawaii. University of Hawaii Press, Honolulu. 166 pp.
Kuligowska, K., H. Lutken, and R. Muller. (2016). Towards development of new ornamental plants: status and progress in wide hybridization. Planta 244: 1-17.
Murashige, T., and F. Skoog. (1962). A revised medium for rapid growth and bioassays with tobacco tissue cultures. Physiol. Plant. 15: 473-497.
National Agricultural Statistics Service. (2019). Census of Horticultural Specialties. Retrieved June 16, 2021, from: https://www.nass.usda.gov/ Publications/AgCensus/2017/Online_Resources/Census_of_Horticulture_Specialties/HORTIC.pdf
Ng, T. B., J. Y. Liu, J. H. Wong, X. J. Ye, S. C. W. Sze, Y. Tong, and K. Y. Zhang. (2012). Review of research on Dendrobium, a prized folk medicine. Appl. Microbiol. Biotechnol. 93: 1795-1803.
Niu, S. C., J. Huang, Q. Xu, P. X. Li, H. J. Yang, Y. Q. Zhang, G. Q. Zhang, L. J. Chen, Y. X. Niu, Y. B. Luo, and Z. J. Liu. (2018). Morphological type identification of self-incompatibility in Dendrobium and its phylogenetic evolution pattern. Int. J. Mol. Sci. 19: 2595.
O'Neill, S. D. (1997). Pollination regulation of flower development. Annu. Rev. Plant Physiol. Plant Mol. Biol. 48: 547-574.
Ockendon, D. (1978). Effect of hexane and humidity on self-incompatibility in Brassica oleracea. Theor. Appl. Genet. 52: 113-117.
OrchidRoots. Retrieved June 16, 2021, from: https://www.orchidroots.com/
Pinheiro, F., D. Cafasso, S. Cozzolino, and G. Scopece. (2015). Transitions between self-compatibility and self-incompatibility and the evolution of reproductive isolation in the large and diverse tropical genus Dendrobium (Orchidaceae). Ann. Bot. 116: 457-467.
Rieseberg, L. H., and S. E. Carney. (1998). Plant hybridization. New Phytol. 140: 599-624.
Singh, R. J. (2017). Plant cytogenetics. CRC press. Boca Raton, London, New York. 528 pp.
Suwabe, K., G. Suzuki, and M. Watanabe. (2010). Achievement of genetics in plant reproduction research: the past decade for the coming decade. Genes Genet. Syst. 85: 297-310.
Takayama, S., and A. Isogai. (2005). Self-incompatibility in plants. Annu. Rev. Plant Biol. 56: 467-489.
Teixeira da Silva, J. A., E. Tsavkelova, T. B. Ng, S. Parthibhan, J. Dobránszki, J. C. Cardoso, M. V. Rao, and S. J. Zeng. (2015). Asymbiotic in vitro seed propagation of Dendrobium. Plant Cell Rep. 34: 1685-1706.
Teixeira da Silva, J. A., S. J. Zeng, R. F. Galdiano, Jr., J. Dobránszki, J. C. Cardoso, and W. A. Vendrame. (2014). In vitro conservation of Dendrobium germplasm. Plant Cell Rep. 33: 1413-1423.
Tsai, C. C. (2012). A new hybrid genus Amenopsis (orchidaceae) derived from the cross between Amesiella and Phalaenopsis. International Symposium on Orchids and Ornamental Plants 1025: 57-60.
Twyford, A. D., and R. A. Ennos. (2012). Next-generation hybridization and introgression. Heredity 108: 179-189.
Udomdee, W., P. J. Wen, C. Y. Lee, S. W. Chin, and F. C. Chen. (2013). Effect of sucrose concentration and seed maturity on in vitro germination of Dendrobium nobile hybrids. Plant Growth Regul. 72: 249-255.
Wang, J., D. Li, F. Shang, and X. Kang. (2017). High temperature-induced production of unreduced pollen and its cytological effects in Populus. Sci. Rep. 7: 1-12.
Whitecross, M., and R. Willing. (1975). Hybridization of incompatible poplars following solvent treatment of stigmas. Experientia 31: 651-653.
Willing, R., and L. Pryor. (1976). Interspecific hybridisation in poplar. Theor. Appl. Genet. 47: 141-151.
Wood, H. P. (2006). The Dendrobiums. ARG Gantner Verlag, Ruggell. 847 pp.
Zheng, S. G., Y. D. Hu, R. X. Zhao, S. Yan, X. Q. Zhang, T. M. Zhao, and Z. Chun. (2018). Genome-wide researches and applications on Dendrobium. Planta 248: 769-784.