摘要
夏蓳(Torenia fournieri Linden)為臺灣夏季重要花壇植物，株型直立不耐低溫，前人利用夏蓳‘Clown Rose’與臺灣原生毛葉蝴蝶草(T. benthamiana Hance)之種間雜交後代(Cb)具有耐低溫及植株生長匍匐性之特點，並經秋水仙素處理獲得Cb5-M612M1。本研究透過選拔Cb5-M612之自交後代(S1)，分析其花朵性狀並建立不同培植體來源搭配疊氮化鈉之微體繁殖技術，期能大量繁殖並開發新品系。
以流式細胞儀檢測Cb及Cb5-M612 之葉片細胞DNA含量，結果顯示Cb5-M612為近四倍體。Cb5-M612經自交後觀察其後代花朵性狀，Cb5-M612S1及Cb5-M612S2側花瓣邊緣鋸齒狀較全緣為顯性，兩族群後代側花瓣全緣與鋸齒後代數量比符合1 : 15 (χ2 = 1.65 <χ20.05,2 = 5.99)，Cb5-M612S1及Cb5-M612S2之唇瓣皆有黃點性狀。Cb5-M612S1花冠長寬出現分離情形，人為選拔大花單株Cb5-M612-S34，其小花梗長度、苞片長度及花冠面長度與夏蓳‘Clown Rose’無顯著差異。
以日/夜溫30/25與15/13 oC處理Cb5-M612S1與夏蓳‘Clown Rose’，並比較植株存活率、葉綠素計讀值、葉片相對傷害值與葉綠素螢光值。結果顯示低溫15/13 oC處理下，Cb5-M612S1皆能維持正常生長，然夏蓳‘Clown Rose’於15/13 oC之存活率僅50%。Cb5-M612S1於低溫15/13 oC仍保持較高之葉綠素計讀值。於30/25 oC及15/13 oC處理下，Cb5-M612S1之葉片細胞膜相對傷害值均較夏蓳‘Clown Rose’低。Cb5-M612S1於30/25與15/13 oC處理之Fv/Fm值無明顯差異，而夏蓳‘Clown Rose’於 15/13 oC處理前三天Fv/Fm值顯著下降，且其三天後之Fv/Fm值較30/25o處理者低。由生長表現、葉綠素濃度、葉片相對傷害值及葉綠素螢光值調查項目顯示，Cb5-M612S1比夏蓳‘Clown Rose’有較佳之低溫(15/13 oC)耐受性。
取Cb5-M612S1不同大小花蕾培植體，置於添加不同濃度NAA與BA之1/2MS培養基，結果顯示花萼稜未發育之花蕾培植體所誘導的芽體數較花萼稜完全發育之花蕾培植體多，以花稜尚未發育階段花蕾培植體培養在0.1 mg‧L-1 NAA 組合2 mg‧L-1 BA平均可獲得較多不定芽數目。
取葉片培植體誘導15天發育至心形期體胚，進行分切繼代至含有不同濃度BA及NAA之1/2MS培養基中，培養四個月後調查體胚發育情形，結果顯示各處理都有癒傷組織形成。每個體胚培植體於不添加任何生長調節劑之培養基中平均可再生6.5個體胚苗且均能發根。於0.1mM NAA組合BA培養基誘導下，體胚苗數目隨BA濃度提高而下降。
取培養四週，長0.5-1.0 cm之葉片誘導之除根體胚苗，沾取不同濃度之IBA與NAA發根劑，結果顯示體胚苗於出瓶四週後之發根率以不沾發根劑之對照組最佳，為93%，平均根數3.9條，主根長達3.6 cm。發根率、平均根數、地上部乾重於IBA與NAA發根劑處理間無顯著差異，地下部乾重以NAA處理者較低。
探討不同疊氮化鈉施用方式及處理部位對Cb5-M612之誘變影響。結果顯示於相同濃度疊氮化鈉處理下，以疊氮化鈉培養基處理者，隨培養週數由1週增加至4週，體胚培植體存活率逐漸降低，以疊氮化鈉溶液浸泡處理者，隨浸泡時間由30 min增加至90 min會造成體胚苗數目逐漸下降。比較花蕾培植體與體胚培植體浸泡疊氮化鈉溶液，結果顯示花蕾培植體之存活率隨處理濃度(1、2及4 mM)及浸泡時間(30、60、90 min)增加而顯著降低，顯示花蕾培植體相對於體胚培植體，藥劑傷害程度較大。經疊氮化鈉藥劑處理獲得內輪花序增生(Paracorolla)、雄蕊瓣化、側花瓣分裂之三種重瓣花類型及一株淡花色之變異株。

Abstract
Wishbone flower (Torenia fournieri Linden) is one of the most important summer bedding plants in Taiwan. Torenai fournieri ‘Clown Rose’ is erect and cold-intolerant. Tetraploid Torenia hybrids from crossing T. fournieri ‘Clown Rose’ with T. benthamiana (Cb) are trailing and low-temperature tolerant, as shown by a previous study. Further colchicine treatments resulted in Cb5-M612M1. The objectives of this study were 1) to investigate flower petal traits of selected self-crossed tetraploid Torenia hybrid progeneies (Cb5-M612S1), 2) to establish micropropagation technique using various explant sources and sodium azide treatments, 3) to select new cultivars/ lines.
Flow cytometric analysis revealed that Cb5-M612M1 was approximately tetraploid. The Cb5-M612M1 was then self-crossed to obtain Cb5-M612S1 and Cb5-M612S2 for the measurements of flower traits. Results showed that serrate edge of lateral petal was dominant to whole edge in S1 and S2 populations and population ratio was approximately 15 : 1 (χ2 = 1.65 <χ20.05,2 = 5.99). All plants had papillae petals with yellow dot in S1 and S2 populations. Corolla size of S1 progeneies showed segregation. The selected large-flowered progeny (Cb5-M612S-34) had similar length of stalks, bracts, and corollas as compared to T. fournieri ‘Clown Rose’ .
Cb5-M612S1 and T. fournieri ‘Clown Rose’ were grown under day/night temperatures of 30/25 and 15/13 oC for 21 days. Cb5-M612S1 at 15/13 oC all survived and maintained high SPAD-502 value, while 50% of T. fournieri ‘Clown Rose’ at 15/13 oC died. Leaf relative injury value in Cb5-M612S1 was lower than T. fournieri ‘Clown Rose’ both under 30/25 and 15/13 oC. Fv/Fm value of Cb5-M612S1 did not differ between temperatures, while T. fournieri ‘Clown Rose’ had significantly decreased Fv/Fm value at first three days at 15/13 oC and thereafter remained lower Fv/Fm values than plants at 30/25 oC. In conclusion, Cb5-M612S1 showed better low temperature (15/13 oC) tolerance than T. fournieri ‘Clown Rose’ according to growth performance, leaf chlorophyll concentration, leaf relative injury, and leaf Fv/Fm value.
Different sizes of flower bud explant in Cb5-M612S1 were cultured on 1/2 MS medium containing various NAA and BA concentrations. Results showed that young flower buds with folded calyx developed more adventitious shoots than those mature flower buds. Young flower buds with folded calyx cultured on 1/2 MS supplemented with 0.1 mg‧L-1 NAA+ 2 mg‧L-1 BA had higher shoot regeneration.
Heart-shaped somatic embryos induced from the leaf explant of Cb5-M612M1 were used for somatic emblings multiplication. Each somatic embryo explant produced an avearge of 6.5 somatic emblings and 100% rooting when cultured without plant regulators. Number of somatic emblings decreased when BA concentration increased from 1 mg‧L-1 to 4 mg‧L-1 in medium supplemented with 0.1 mg‧L-1 NAA.
After 4 weeks in vitro culture, somatic emblings with 0.5-1.0 cm length were used for ex vitro rooting experiments. Results showed that somatic emblings without rooting hormone had highest rooting percentage. There were no significant differences in rooting percentage, root number, and shoot dry weight between IAA and NAA treatments. NAA treatments resulted in lower root dry weight.
Sodium azide (SA) treatments and explant source were investigated for the mutation in Cb5-M612M1. Results showed that somatic embryo explant survival percentage decreased when SA medium culture duration increased from 1 to 4 weeks, and somatic emblings number per explant decreased with increasing SA solution soaking duration from 30 to 90 min. Flower bud explants were more sensitive to sodium azide solution soaking treatment than somatic embryo explant. Three double-flowered mutations (paracorolla, petaloid of stamen, and dissection of lateral petal), and one pale flower color mutation were obtained after sodium azide treatments.

目錄
摘要 i
Abstract iii
表目錄 ix
圖目錄 x
前言(Introduction) 1
前人研究(Literature Review) 4
一、倒地蜈蚣屬重要種源 4
二、夏蓳育種史與品種選育方向 5
三、耐寒性狀導入及檢測 6
四、植物色素生合成系統 8
五、夏蓳花色素組成與花色調控 9
六、雜交後代之花朵性狀表現 10
七、重瓣花分類及其調控 11
八、影響重瓣花的外在因素 14
九、多倍體化育種途徑及應用 14
十、微體繁殖再生系統 15
十一、花卉誘變育種 19
材料與方法 (Material and Methods) 22
試驗一、Cb5-M612稔性篩選及其DNA含量分析 22
試驗二、Cb5-M612S1與其來源親本花朵性狀比較及大花單株選拔 23
試驗三、Cb5-M612S1及Cb5-M612S2花朵性狀調查 23
試驗四、日/夜溫對Cb5-M612S1與夏蓳‘Clown Rose’ 葉綠素螢光值、葉綠素計讀值、相對生長速率與葉片相對傷害值之影響 24
試驗五、NAA與BA對Cb5-M612不同大小之花蕾培植體誘導再生不定芽之影響 25
試驗六、NAA與BA對Cb5-M612葉片衍生體胚誘導再生之影響 26
試驗七、IBA與NAA濃度對Cb5-M612體胚苗瓶外發根之影響 26
試驗八、疊氮化鈉培養基濃度與處理時間對Cb5-M612體胚存活率、再生體胚苗數目與誘變率之影響 27
試驗九、疊氮化鈉浸泡濃度與時間對Cb5-M612體胚培植體存活率、再生體胚苗數目與誘變率之影響 28
試驗十、疊氮化鈉浸泡濃度與時間對Cb5-M612花蕾再生不定芽數目、芽體再生率與誘變率之影響 28
結果(Results) 30
試驗一、Cb5-M612稔性篩選及其DNA含量分析 30
試驗二、Cb5-M612S1與其來源親本花朵性狀比較及大花單株選拔 30
試驗三、Cb5-M612S1及Cb5-M612S2花朵性狀調查 30
試驗四、日/夜溫對Cb5-M612S1與夏蓳‘Clown Rose’ 葉綠素螢光值、葉綠素計讀值、相對生長速率與葉片相對傷害值之影響 32
試驗五、NAA與BA對Cb5-M612不同大小之花蕾培植體誘導再生不定芽之影響 32
試驗六、NAA與BA對Cb5-M612葉片衍生體胚誘導再生之影響 33
試驗七、IBA與NAA濃度對Cb5-M612體胚苗瓶外發根之影響 33
試驗八、疊氮化鈉培養基濃度與處理時間對Cb5-M612體胚存活率、再生體胚苗數目與誘變率之影響 34
試驗九、疊氮化鈉浸泡濃度與時間對Cb5-M612體胚培植體存活率、再生體胚苗數目與誘變率之影響 34
試驗十、疊氮化鈉浸泡濃度與時間對Cb5-M612花蕾再生不定芽存活率、芽體再生數目與誘變率之影響 35
討論(Discussion) 69
參考文獻(References) 77
附錄 (Appendix) 93

參考文獻(References)
呂廷森、立澤文見. 2012. 夏菫花色與花青素色素組成之比較研究. 臺灣園藝 57:346.
朱德民. 1993. 植物與環境逆境. 植物與溫度逆境. 國立編譯館. p.87-144.
江純雅、張惠媜、黃倉海、黃敏展、趙玉真、馮莉真、蕭伊芸、朱建鏞. 2006. 利用組織培養之誘變育種. p.75-91. 花卉育種研討會專刊.中興大學園藝學系出版.
朱建鏞、江純雅. 2004. 菊花組織培養之誘變育種. 植物種苗6:13-19.
沈再木、黃光亮、張平順. 2001. 疊氮化鈉對夜來香品系誘變效應之初報. 中國園藝 47:129-135.
郭乃瑋. 2011. 文心蘭體胚發生及疊氮化鈉誘變之研究. 國立中興大學農藝學系碩士論文. 臺中.
林添成. 1992. 夏蓳的體胚發生與器官形成. 國立臺灣大學園藝學系碩士論文. 臺北.
徐碧徽. 2004. 短期貯藏、母本遮陰與氮鉀肥濃度對菊花插穗發根與生長之影響. 國立臺灣大學園藝學系碩士論文. 臺北.
陳治官、黃真生. 1984. 疊氮化鈉對水稻台農67號之誘變效果. 中華農業研究 33:345-353.
陳錦木. 2011. 臺灣花壇植物產業現況與展望. 綠色城市與花卉產業國際研討會論文集. 國立臺灣大學園藝學系. 臺北.
高典林. 2009. 現代作物育種學. 藝軒圖書出版社, 台北.
張惠媜、朱建鏞. 2005. 非洲菊利用花蕾培養處理疊氮化鈉進行誘變育種. 植物種苗7:23-33.
楊喬雯. 2013. 美鐵芋器官發生與體胚發生再生系統之建立. 國立嘉義大學園藝學系碩士論文. 嘉義.
趙印泉、劉青林. 2009. 重瓣花的形成機理及遺傳特性研究進展. 西北植物學報 4:832-841.
黃柄龍. 2011. 疊氮化鈉與γ射線對蜻蜓屬及擎天屬觀賞鳳梨器官分化過程誘變效應之初探. 高雄區農業改良場研究彙報 21:18-27.
黃筱&;#23169;. 2008. 粗肋草花序培養、不定芽再生繁殖及瓶內馴化. 國立臺灣大學園藝學系碩士論文. 臺北.
謝順景、謝順鑫. 1981. 誘變育種學. 國立編譯館, 臺北.
蘇姬泰. 2009. 利用疊氮化鈉培養基誘變長壽花. 國立中興大學園藝學系碩士論文. 臺中.
魏子耀. 2010. 夏蓳之花朵性狀遺傳與遠緣雜交障礙. 國立臺灣大學園藝學系碩士論文. 臺北.
郭孟樺. 2012. 夏蓳與毛葉蝴蝶草種間雜交胚之拯救與多倍體化. 國立臺灣大學園藝暨景觀學系碩士論文. 臺北.
Abul-Soad, A.A. 2011. Micropropagation of data palm using inflorescence explants, p.91-117. In: S.M. Jain, J.M. Al-Khayri, and D.V. Jonson (eds.). Data palm biotechnology. Springer Netherlands. New York.
Ahn, Y.J., L. Vang, T. A. Mckeon, G.Q. Chen. 2007. High-frequency plant regeneration through adventitious shoot formation in castor (Ricinus communis L.). In Vitro Cell. Dev. Biol. Plant 43:9-15.
Allum, J.F., D.H. Bringloe, and A.V. Roberts. 2007. Chromosome doubling in a Rosa rugosa Thunb. hybrid by exposure of in vitro nodes to oryzalin: The effects of node length, oryzalin concentration and exposure time. Plant Cell Rpt. 26: 1977-1984.
Al-Qurainy, F., and S. Khan. 2009. Mutagenic effects of sodium azide and its application in crop improvement. World Appl. Sci. J. 6:1589-1601.
Aida, R., K. Yoshida, T. Kondo, S. Kishimoto, and M. Shibata. 2000a. Copigmentation gives bluer flowers on transgenic torenia plants with the antisense dihydroflavnonol-4-reductase gene. Plant Sci. 160:49-56.
Aida, R., S. Kishimoto, Y. Tanaka, and M. Shibata. 2000b. Modification of flower color im torenia (Torenia fournieri Lind.) by genetic transformation. Plant Sci. 153:33-42.
Aida, R., M. Komano, M. Saito, K. Nakase, and K. Murai. 2008. Chrysanthemum flower shape modification by suppression of Chrysanthemum-AGAMOUS gene. Plant Biotechol. 25:55-59.
Almeida, J., M. Rocheta, and L., Galego. 1997. Genetic control of flower shape in Antirrhinum majus. Dev. 124:1387-1392.
Anderson, J.A., M.G. Kenna, and C.M. Taliaferro. 1988. Cold hardiness of ‘Midiron’ and ‘Tifgreen’ bermudagrass. HortScience 23:748-450.
Andrews, J.R., M.J. Fryer, and N.R. Baker. 1995. Characterization of chilling effects on photosynthetic performance of maize crops during early season growth using chlorophyll fluorescence. J. Expt. Bot. 46:1195-1203.
Anis, M., A. Varshney, and I. Siddique. 2010. In vitro clonal propgagation of Balanites aegyptiaca L. Agrofor. Syst. 78:151-158.
Aro, E.-M., T. Hundal, I. Carlberg, and B. Andersson. 1990. In vitro studies on light-induced inhibition of photosystem II and D1-protein degradation at low emperatures. Biochim. Biophys. Acta. 1019:269–275.
Australia Government. 2006. The biology of Torenia spp. (Torenia). Office of the Gene Technology Regulator, Department of Health and Ageing. 31 May 2009. < http://www.ogtr.gov.au/ >.
Beck, M.J. and N.D. Camper. 1991. Shoot regernation from petunia leaf discs as a function of explant size, configuration and benzyladenine exposure. Plant Cell Tiss. Org. Cult. 26:101-106.
Belesky, D.P., H.D. Perry, W.R. Windham, E.L. Mathias, and J.M. Fedders. 1991. Productivity and quality of bermudagrass in a cool temperature environment. Agron. J. 83:810-813.
Benkirane, H., K. Sabounji, A. Chlyah, and H. Chlayh. 2000. Somatic embryogenesis and plant regeneration from fragment of immature inflorescence and coleoptiles of durum wheat. Plant Cell Tiss. Org. Cult. 61:107-113.
Benmahioul, B., N. Dorion, M. Kaid-Harche, F. Daguin. 2012. Micropropagation and ex vitro rooting of pistachio (Pistacia vera L.). Plant Cell Tiss. Org. Cult. 108:353-358.
Bhate, R.H. 2001. Chemically induced floral morphological mutations in two cultivars of Ipomoea purpurea (L.) Roth. Sci. Hortic. 88:133-145.
Bhatia, N.P., P. Bhatia, and N. Ashwath. 2002. Ex vitro rooting of micropropagated shoots of Stackhousia tryonii. Biol. Plant. 45:441-444.
Birth, G.S. and G.R. McVey. 1968. Measuring the color of growthing turf with a reflectance spectrophotometer. Agron. J. 60:640-642.
Blom, T. 1980. Rose research in Veinland, Canada. Bull. Roses Inc. June. p. 45-46.
Bowman, J.L., D.R. Smyth, and E.M. Meyerowitz. 1989. Genes directing flower development in Arabidopsis Plant Cell 1:37-52.
Bradley, D., R. Carpenter, H. Sommer, N. Hartley, and E. Coen. 1993. Complementary floral homeotic phenotypes result from opposite orientations of a transposon at the plena locus of Antirrhinum. Cell 72:85-95.
Bridgen, M.P., M.Z. Hadi, and M.S. Barreto. 1992. A laboratory exercise to demonstrate direct and indirect shoot organogenesis from leaves of Torenia fournieri. HortTechnology 4:320-322.
Broertje, C. and A.M. van Harten. 1988. Application of mutation breeding methods in the improvement of vegetatively propagated crops. Elsevier, New York.
Campos, P.S., V. Quartin, J. C. Ramalho, and M.A. Nunes. 2003. Electrolyte leakage and lipid degradation account for cold sensitivity in leaves of Coffea sp. Plants. J. Plant Physiol. 160:283-292.
Caswell, K.L., N.L. Leung, and R.N. Chibbar. 2000. An efficient method for in vitro regeneration form immature inflorescence explants of Canadian wheat cultivars. Plant Cell Tiss. Org. Cult. 60:69-73.
Cheng, X.Y. and M.W. Gao. 1988. Biological and genetic effects of combined treatments of sodium azide, gamma rays and ems in barley. Environ. Expt. Bot. 28:281-288.
Cheng, X., S. Chen, F. Chen, W. Fang, Y. Deng, and L. She. 2010. Interspecific hybrids between Dendranthema morifolium (Ramat.) Kitamura and D. nankingense (Nakai) Tzvel. achieved using ovary rescue and their cold tolerance characteristics. Euphytica 172:101-108.
Chu, C. and T.M. Lee. 1989. The relationship between ethylene biosynthesis and chilling tolerance in seedlings of rice (Orzya sativa L.). Bot. Bull. Acad. Sin. 30:263-273.
Chugh, S., S. Guha, and I.U. Rao. 2009. Micropropagation of orchids: A review on the potential of different explants. Scientia Hort. 122:507-520.
Coen, E.S. and E. M. Meyerowitz. 1991. The war of the whorls: Genetic interactions controlling flower development. Nature 353:31-37.
Davies, M.E. and M.M. Dale. 1979. Factors affecting in vitro shoot regeneration on leaf discs of Solanum laciniatum. J. Plant Physiol. 92:51-60.
Debergh, P.C., and R.H. Zimmerman. 1991. Acclimatization of micropropagated plants to the greenhouse and filed, p.71-93. In: Preece, J.E., and E.G. Sutter (eds.). Microprogagation. Gent Univ. State, Agriculture U.S.D.
De Klerk, G.J., W.V.D. Krieken, and J.C. De Jong. 1999. Review: The formation of adventitious roots: New concepts, new possibilities. In Vitro Cell Dev. Biol. 35:189-199.
Delph, L.F., J.L. Gehring, F.M. Frey, A.M. Arntz, and M. Levri. 2004. Genetic consraints on floral evolution in a sexually dimorphic plant revealed by artificial selection. Evolution 58:1936-1946.
Deng, Y., S. Chen, F. Chen, X. Cheng, and F. Zhang. 2011. The embryo rescue derived intergeneric hybrid between chrysanthemum and Ajania przewalskii shows enhanced cold tolerance. Plant Cell Rpt. 30: 2177-2816.
Dhooghe, E., K.V. Laere, T. Eeckhaut, L.Leus, J.V. Huylenbroeck. 2011. Mitotic chromosome doubling of plant tissues in vitro. Plant Cell Tiss. Org. Cult. 104: 359-373.
Donnison, I.S. and D. Francis. 1994. Experimental control of floral reversion in isolated shoot apices of the long-day plant Silene coeli-rosa. Physiol. Plant. 92:329-335.
Dwyer, L.M., M. Tollenaar, and L. Houwing. 1991. A nondestructive method to monitor leaf greenness in corn. Can. J. Plant Sci. 71:505-509.
Endo, T. 1962. Inheritance of anthocyanin concentrations in flowers of Torenia fournieri. Jpn. J. Genet. 37:284-290.
Galbraith, D.W., K.R. Harkins, J.M. Maddox, N.M. Ayres, D.P. Sharma, and E. Firoozadaby. 1983. Rapid flow cytometric analysis of the cell cycle in intact plant tissues. Science 220: 1049-1051.
Ganesan, M., P. Bhanumathi, and N. Jayabalan. 2005. Mutagenic effect of sodium azide on somatic embryo regeneration and root growth of cotton (Gossypium hirsutum L. cv. SVPR2). J. Agric. Technol. p:365-379.
Garrod, J.F. and G.P. Harris. 1974. Studies on glasshouse carnation: Effects of temperature and growth substances on petal number. Ann. Bot. 38:1025-1033.
Gaspar, I. and G. Butanaru. 1985. Triticale O noua. Ed. Academiei Romane, Bucresti.
Gesch, R.W. and J.L. Heilman. 1999. Response of photosynthesis and phosphorylation of the light-harvesting complex of photosystem II to chilling temperature in ecologically divergent cultivars of rice. Environ. Expt. Bot. 41:257-266.
Galen, C. 1996. Rates of floral evolution: Adaptation to bumblebee pollination in an alpine wildflower, Polemmonium viscosum. Evolution 50:120-125.
Guohua, M. and G. Wu, and E. Bunn. 2007. Somatic embryogenesis and adventitious shoot formation in Burma reed (Neyraudia arundinacea Henr.). In Vitro Cell. Dev. Biol. 43:16-20.
Han, D.S. and Y. Niimi. 2008. Fertility restoring of interspecific hybrids between Lilium nobilissimum and L. regale by chromosome doubling. Acta. Hort. 766:421-426.
Hammett, K.R.W., B.G. Murray, K.R. Markham, and I.C. Hallett. 1994. Interspecific hybridization between Lathyrus odoratus and L. belinensis. Intl. J. Plant Sci. 155:763-771.
Hess, D. 1971. Chemogenetical investigations on the synthesis of flower colour substances in Torenia species (Scrophulariaceae). Biochem. Physiol. Pflanzen. 162:S386-S389.
Higashiyama, T., R. Inatsugi, S. Sakamoto, N. Sasaki, T. Mori, H. Kuroiwa, T. Nakada, H. Nozaki, T. Kuroiwa, and A. Nakano. 2006. Species preferentiality of the pollen tube attractant derived from the synergid cell of Torenia fournieri. Plant Physiol. 142:481-491.
Hsieh, T.H. and K.C. Yang. 2002. Revision of Torenia L. (Scrophulariaceae) in Taiwan. Taiwania 47:281-289.
Hyun, S.K. 1976. Interspcific hybridization in pines with the special reference to Pinus rigida × teada. Silvae Genet. 25:188-191.
Ikawa, K. 2005. U.S. Patent PP16055 P3: Torenia plant named ‘Sunrenilapa’. United State Patent &; Trademark office.
Ishizaka, H. and J. Uematsu. 1995. Amphidiploids between Cyclamen persicum Mill. and C. purpurascens Mill. induced by treating ovules with colchicines in vitro and sesquidiploids between the amphidiploid and parental species induced by conventional crosses. Euphytica 86:211-218.
Iwaki, K. 2005a. U.S. Patent PP15560: Torenia plant named ‘Sunrenilahopas’. United State Patent &; Trademark Office.
Iwaki, K. 2005b. U.S. Patent PP15684: Torenia plant named ‘Sunrenilamu’. United State Patent &; Trademark Office.
Iwaki, K. 2005c. U.S. Patent PP16055: Torenia plant named ‘Sunrenilapa’. United State Patent &; Trademark Office.
Johnson, G.V. 1974. Simple procedure for quantitative analysis of turfgrass color. Agron. J. 66:457-459.
Kantia, A., S.L. Kothari. 2002. High efficiency adventitious shoot bud formation and plant regeneration from leaf explants of Dianthus chinensis L. Scientia Hort. 96:205-212.
Katsumoto, Y., M. Fukuchi-Mizutani, Y. Fukui, F. Brugliera, T.A. Holton, M. Karan, N. Nakamura, K. Yonekura-Sakakibara, J. Togami, A. Pigeaire, G.Q. Tao, N. S. Nehra, C.Y. Lu, B.K. Dyson, S. Tsuda, T. Ashikari, T. Kusumi, J.G. Mason, and Y. Tanaka. 2007. Engineering of the rose flavonoid biosynthetic pathway successfully generated blue-hued flowers accumulationg delphinidin. Plant Cell Physiol. 48: 1589-1600.
Karnok, K.J. and J.B. Beard. 1983. Effect of gibberellic acid on the CO2 exchange rates of bermudgrass and augustinegrass when exposed to chilling temeratures. Crop Sci. 23:514-516.
Kermani, M.J., V. Sarasan, A.V. Roberts, K. Tokoya, J. Wentworth, and V.K. Sieber. 2003. Oryzalin-induced chromosome doubling in Rosa and its effect on plant, morphology and pollen viability. Theor. Appl. Genet. 107:1195-1200.
Ketchersid, M.L. and M.G. Merkle. 1976. Dissipation and phytotoxicity of sodium azide in soil. Weed Sci. 24:312-315.
Khalatkar, A.S. and S.G. Kashikar. 1980. Sodium azide mutagenicity in Petunia hybrida. Mutation Res. 79:81-85.
Khawaja, H.I.T. 1988. A new interspecific Lathyrus hybrid to introduce the yellow flower character into sweet pea. Euphytica 37:69-75.
Kikuchi, S., H. Tanaka, T. Shiba, M. Mii, and H. Tsujimoto. 2006. Genome size, karyotype, meiosis and a novel extra chromosome in Torenia fournieri, T. baillonii and their hybrid. Chromosome Res. 14:665-672.
Kikuchi, S., H. Kino, H. Tanaka, and H. Tsujimoto. 2007a. Pollen tube growth in cross combinations between Torenia fournieri and fourteen related species. Breed. Sci. 57:117-122.
Kim M.S., N.B. Klopfenstein, and B.M. Cregg. 1988. In vitro and ex vitro rooting of micropropagated shoots using three green ash (Fraxinus pennsylvanica) clones. New Forests 16:43-57.
Kiruki, S., L.A. Onek, and M. Limo. 2006. Azide-based mutagenesis suppresses Striga hermonthica seed germination and parasitism on maize varieties. African J. Biotechnol. 5:866-870.
Kleinhof, A., C. Sander, R.A. Nilan, and C.P. Konzak. 1974. Polyploid and induced mutations in plant breeding. I.A.E.A., Vienna, Austria.
Kleinhofs, A., A.L. Hodgdon, W.M. Owais, and R.A. Nilan. 1984. Effectiveness and Safety of sodium azide mutagenesis. Appl. Sci. 15:53-58.
Kobayashi, N., S. Yamashita, K. Ohta, and T. Hosoki. 2008. Morphological characteristics and their inheritance in colchicines-induced Salvia polyploids. J. Jpn. Soc. Hort. Sci. 77:186-191.
Koroch, A., H.R. Juliani, J. Kapteyn, and J.E. Simon. 2002. In vitro regeneration of Echinacea purpurea from leaf explants. Plant Cell Tiss. Org. Cult. 69:79-83.
Koyama, T., M. Ohme-Takagi, and F. Sato. 2011. Generation of serrated and wavy petals by inhibition of the acivity of TCP transcription factors in Arabidopsis thaliana. Plant Signal. Behav. 6:697-699.
Lendvai, G. and D. Levin. 2003. Rapid response to artificial selection on flower size in Phlox. Heredity 90:336-342.
Lehtila, K. and K.H. Brann. 2007. Correlated effects of selection for flower size in Raphanus raphanistrum. Can. J. Bot. 85:160-166.
Lim, C.C. and R. Arora. 1998. Comparing gompertz and Richards functions to estimate freezing injury in Rhododendron using electrolyte leakage. J. Amer. Soc. Hort. Sci. 123:246-251.
Lim, C.S., S.M. Kang, and J.L. Cho. 2007. Bell peper (Capsicum annuum L.) fruits are susceptible to chilling injury at the breaker stage of ripeness. HortScience 42:1659-1664.
Lord, E.M. and K.J. Eckard. 1987. Shoot development in Citrus sinensis L.(Washington Navel orange). II. Alteration of developmental fate of flowering shoot after GA3 treatment. Bot. Gaz. 148:17-22.
Lu, C. and M.P. Bridgen. 1996. Effects of genotype, culture medium and embryo developmental stage on the in vitro responses from ovule cultures of interspecific hybrids of Alstroemeria. Plant Sci. 116:205-212.
Lu, C. and M.P. Bridgen. 1997. Chromosome doubling and fertility study of Alstroemeria aurea × A. caryophyllaea. Euphytica 94:75-81.
Mallikarjuna. N. 1999. Ovule and embryo culture to obtain hybrids from interspecific incompatible pollinations in chickpea. Euphytica 110:1-6.
Markhart, S., J.M. Kinet, and J. Bouharmont. 1996. Nacl-induced senescence in leaves of rice (Oryza sativa L.) cultivars differing in salinity resistance. Ann. Bot. 78:389-398.
Maluszynski, M., K. Nichterlein, L. van Zanten and B. S. Ahloowalia. 2000. Officially released mutant varieties- the FAO/IAEA Database. Mut. Breed. Rev. 12:1-84.
Mauro, S., P. Dainese, R. Lannoye, and R. Bassi. 1997. Cold-resistant and cold-seneitive maize lines differ in the phosphlorylation of the photosystem II subunit, CP29. Plant Physiol. 115:171-180.
Mensah, J.K., P.A. Akomeah, and E.O. Ekpekurede. 2005. Gama irradiation induced variation of yield parameters in cowpea (Vigna unguiculata L. Walp.). Global J. Pure Appl. Sci. 11:327-330.
Miyazaki, K. 1999. U.S. Patent PP11131: Torenia plant named ‘Sunrenidibu’. United State Patent &; Trademark Office.
Miyazaki, K. 2001. U.S. Patent PP12105: Torenia plant named ‘Sunrenilabu’. United State Patent &; Trademark Office.
Miyazaki, K. 2002. Torenia plant named ‘Sunrenihopas’. Plant Variety Protection, Ministry Agric. For. Fish.
Miyazaki, K. 2003. U.S. Patent PP14302: Torenia plant named ‘Sunrenirirepa’. United State Patent &; Trademark Office.
Miyazaki, K., K. Suzuki, K. Iwaki, T. Kusumi, T. Abe, S. Yoshida, and H. Fukui. 2006. Flower pigment mutations induced by heavy ion beam irradiation in an interspecific hybrid of Torenia. Plant Biotechnol. 23:163-167.
Molina-Cano, J.L., J.P. Simian, A. Spena, A.M. Perez-Vendrell, S. Dorsch, D. Rubiales, J.S. Swanston, and A. Jahoor. 2003. Mildew-resistant mutants induced in North American two-and six-rowed malting barely cultivars. Theor. Appl. Genet. 107:1278-1287.
Murashige, T. and F. Skoog. 1962. A revised medium for rapid growth and bio-assays with tobacco tissue culture. Physiol. Plant. 15:473-497.
Nagase, Y. 1997. U.S. Patent PP10119: Torenia plant named ‘Sunrenibu’. United State Patent &; Trademark Office.
Nakamura, N., M. Fukuchi-Mizutani, K. Miyazaki, K. Suzuki, and Y. Tanaka. 2006. RNAi suppression of the anthocyanidin synthase gene in Torenia hybrid yields white flowers with higher frequency and better stability than antisense and sense suppression. Plant Biotechnol. 23:13-17.
Nakamura, N., M. Fukuchi-Mizutani, Y. Fukui, K. Ishiguro, K. Suzuki, H. Suzuki, K. Okazaki, D. Shibata, and Y. Tanaka. 2010. Generation of pink flower varieties from blue Torenia hybrid by redirecting the flavonoid biosynthetic pathway from delphinidin to pelargonidin. Plant Biotechnol. 27:375-383.
Narumi, T., R. Aida, T. Koyama, Hi. Yamaguchi, K. Sasaki, M. Shikata, M. Nakayama, M. Ohme-Takagi, and N. Ohtsubo. 2011. Arabidopsis chimeric TCP3 repressor produces novel floral traits in Torenia fournieri and Chrysanthemum morifolium. Plant Biotechnol. 28:131-140.
Niki, T., M. Hirai, A. Kanno, and T. Nishijima. 2012. Role of floral homeotic genes in the morphology of forchlorfenuron-induced paracorollas in Torenia fournieri Lind. J. Jpn. Soc. Hort. Sci. 81:204-212.
Niki, T., T. Mahesumu, T. Niki, and T. Nishijima. 2013. Localized high expression of type-A response regular and cytokinin oxidase/dehydrogenase genes in relation to forchlorfenuron-induced changes in flower morphology in Torenia fournieri Lind. J. Jpn. Soc. Hort. Sci. 82:69-77.
Niki, T., R. Aida, T. Niki, and T. Nishijima. 2013. Effect of localized promotion of cytokinin biosynthesis on flower morphology in flower buds of Torenia fournieri Lind. J. Jpn. Soc. Hort. Sci. 82:328-336.
Nilan, R.A., E.G. Sideris, A. Kleninhofs, and C.F. Konzak. 1973. Azide-A potent mutagen. Mutat. Res. 17:142-144.
Nirmal, K.S. and C.B. Sehgal. 1999. Micropropgagation of ‘Holly Basil’ (Ocimum sanctum Linn.) from young inflorescence of mature plants. Plant Growth Regulat. 29:161-166.
Nishijima, T. and K. Shima. 2006. Change in flower morphology of Torenia fournieri Lind. induced by forchlorfenuuron application. Scienta Hort. 109:254-261.
Nitasaka, E. 2003. Insertion of an En/Spm-related transposable element into a floral homeotic gene DUPLICATED causes a double flower phenotype in the Japanses moring glory. Plant J. 36:522-531.
Offord, C.A. and L.C. Campbell. 1992. Micropropagation of Telopea speciosissima R. Br. (Proteaceae). 2: Rhizogenesis and acclimatization to ex vitro conditions. Plant Cell Tiss. Org. Cult. 29:223-230.
Papadopoulou, K., R.E. Melto, M. Leggett, M.J. Daniels, and A.E. Osbourn. 1996. Compromised diseased resistance in saponin deficient plant. Proc.Nat. Acad. Sci. USA. 96:12923-12928.
Park, Y.G. and S.H. Son. 1988. Regeneration of plantlets from cell suspension culture derived callus of white poplar (Populus alba L.). Plant Cell Rpt. 7:567-570.
Pierik, R.L.M. 1987. In vitro culture of higher plants. Martinus Nijhoff Publisher, Dordrecht.
Pintos, B., J.P. Martin, M.L. Centeno, N. Villalobos, H. Guerra, L. Martin. 2002. Endogenous cytokinin levels in embryogenic and non-embryogenic calli of Medicago arborea L. Plant Sci. 163:955-960.
Pospi&;#353;ilova, J., I. Ticha, P. Kadlecek, D. Haisel, and &;#352;. Plzakova. 1999. Acclimatization of micropropagated plants to ex vitro conditions. Biol. Plant. 42:481-497.
Qu, R. and A. Chaudhury. 2001. Improved young inflorescence culture and regeneration of ‘Tifway’ bermudgrass (Cynodon transvaalensis ×dactylon). Intl. Turf. Soc. Res. J. 9:198-201.
Rajib, R. and J. Tah. 2011. Assessment of chemical mutagenic effects in mutation breeding programme for M1 generation of carnation Dianthus caryophyllus. Res. Plant Biol. 1:23-32.
Raman, K., and R.I. Greyson. 1978. Differential effects of kinetin on acidic ethyl acetate-soluble and highly water-soluble gibberellin fractions in ‘Single’ and ‘Double’ Nigella damascene L. Expt. Bot. 29:141-145.
Ramsey, J. and D.W. Schemske. 1988. Pathways, mechanisms, and rates of polyploidy formation in flowering plants. Annu. Rev. Ecol. System. 29:467-501.
Rao, G.U., M. Lakshmikumraran, and K.R. Shivanna. 1996. Production of hybrids, amphiplids and backcross progenies between a cold-tolerant wild species, Erucastrum abyssinicum and crop brassicas. Theor. Appl. Genet. 92:786-790.
Rapacz, M., D. Gasior, Z. Zwierzykowski, A. Lesniewska-Bocianowska, M.W. Humphreys, and A.P. Gay. 2004. Changes in cold tolerance and the mechanisms of acclimation of photosystem II to cold hardening generated by anther culture of Festuca pratensis × Lolium multiflorum cultivars. New Phytol. 162:105-114.
Reynolds, J. and J. Tampion. 1983. Double flowers: A scientific study. Scientific and Academic Editions, New York.
Roy, S. and A.K. Biswas. 2005. Isolation of a white flowered mutant through seed culture in Spathoglottis plicata Blume. Cytologia 70:1-6.
Salvi, N.D., L. George, and S. Eapen. 2000. Direct regeneration of shoots from immature inflorescence cultures of turmeric. Plant Cell Tiss. Org. Cult. 62:235-238.
Sarma, N.P., A. Patnaik, and P.J. Jachuck. 1979. Concentration and soaking time on induced chlorophyll mutation frequency. Environ. Expt. Bot. 19:117-21.
Skoog, F. and C.O. Miller. 1957. Chemical regulation of growth and organ formation in plant tissue cultures in vitro. Symp. Soc. Expt. Biol. 11:118-131.
Somleva, M.M., V. Kapchina, V. Alexieva, and E. Golovinsky. 1995. Anticytokinin effects on in vitro response of embryogenic and nonembryogenic genotypes of Dactylis glomerata L. Plant Growth Regul. 16:109-112.
Souza, V.C. and A.M. Giulietti. 2009. Scrophulariaceae sensu lato native from Brazil. Pesquisas Bot. 60:7-288.
Suzuki, K., H.M. Xui, Y. Tanaka, Y. Fukui, M. Fukuchi-Mizutani, Y. Murakami, Y. Katsumoto, S. Tsuda, and T. Kusumi. 2000. Flower color modification of Torenia hybrid by cosupression of anthocyanin biosynthesis genes. Mol. Breeding 6:239-246.
Suzuki, K., K. Miyazaki, Y. Tanaka, and K. Iwaki. 2002a. Torenia plant named ‘Sunreniho’. Plant Variety Protection, Ministry Agric. For. Fish. Japan.
Suzuki, K., K. Miyazaki, Y. Tanaka, and K. Iwaki. 2002b. Torenia plant named ‘Sunrenibuho’. Plant Variety Protection, Ministry Agric. For. Fish. Japan.
Tanaka, Y. and A. Ohmiya. 2008. Seeing is believing: engineering anthocyanin and carotenoid biosynthenic pathways. Curr. Opin. Biothechnol. 19:190-197.
Tandon, S.L. and K. Bhutani. 1965. Morphological and cytological studies of colchicines-induced tetraploids in Torenia fournieri Lind. Genetica 36:439-445.
Tamura, Y. and K. Miyazaki. 1999. U.S. Patent PP10843: Torenia plant named ‘Sunrenilabu’. United State Patent &; Trademark Office.
Thakur, S. and P.S. Ganapathy. 1978. Morphogenesis of organ differtiation in Bacopa monnieri stem cultures. Indian J. Exp. Biol. 2:514-516.
Tomosone, S. and D. Gertnere. 2003. In vitro shoot regeneration from flower and leaf explants in Rhododendron. Biol. Plant. 46:463-465.
Tomar, U.K. and S.C. Gupta. 1988. In vitro plant regeneration of leguminous tress (Albizia spp.). Plant Cell Rpt. 7:385-388.
Tooke, F. and N.H. Battey. 2000. A leaf-derived signal is a quantitative determination of floral form in Impatiens. Plant Cell 12:1837-1847.
Topoonyanout, N., R. Ampawan, and P.C. Debergh. 2000. Reversion of Limonium hybrid ‘Misty Blue’ inflorescence development and its applicability in micropropagation. Scientia Hort. 83:283-299.
Trejgell, A., M. Bednarek, and A. Tretyn. 2009. Micropropgation of Carlina Acaulis L. Acta. Biol. Cracov. Bot. 51:97-103.
Tychonievich, J. and R.M. Warner. 2011. Interspecific crossability of selected Salvia species and potential use for crop improvement. J. Amer. Soc. Hort. Sci. 136:41-47.
Weijer, J. 1959. The interaction of gibberellic acid and indoleacetic in Impatiens. Science 129:896-897.
Van Harten, A.M. 1998. In vitro techniques for mutation breeding. p. 163-203. In: A.M. van Harten (eds.). Mutation Breeding: Theory and Practical Applications. Cambridge University Press, United Kingdom.
Venglat, S.P. and V.K. Sawhney. 1996. Benzylaminopurine induces phenocopies of floral meristem and organ identity mutants in wild-type Arabidopsis plants. Planta 198:480-487.
Verdeil, J.L., Huet, F. Grosdemange, and J. Buffard-Morel. 1994. Plant regeneration form cultured immature inflorscences of coconut (Cocos nucifera L.): Evidence for somatic embryogenesis. Plant Cell Rpt. 13:218-221.
Verheul, M.J., P.R. van Hassel, and P. Stamp. 1995. Comparsion of maize inbred lines differing in low temperature tolerance: Effect of acclimation at suboptimal temperature on chloroplast functioning. Ann. Bot. 76:7-14.
Williams, E.G. and G. Maheswaram. 1986. Somatic embryogenesis: Factors influcencing coordinated behavior of cells as an embryogenic group. Ann. Bot. 57:443-462.
Worley, A. C. and S. C. Barrett. 2000. Evolution of floral display in Eichhornia paniculata (Pontederiaceae): Direct and correlated responses to selection on flower size and number . Evolution 54:1533-1545.
Wyka, T.P., M. Hamerska, and M. Wroblewska. 2006. Organogenesis of vegetative shoots from in vitro cultured flower buds of Mammillaria albicoma (Cactaceae). Plant Cell Tiss. Org. Cult. 87:27-32.
Yabuya, T. 1985. Amphidiploids between Iris laevigata Fisch, and I. ensata Thunb. induced through in vitro culture of embryos treated with colchicines. Jpn. J. Breed. 35:136-144.
Yamazaki, T. 1985. A revision of the genera Limnophila and Torenia from Indochina. J. Fac. Sci. Imp. Univ. Tokyo Sec. III 13:575-624.
Ziv, M. and H. Lilien-Kipnis. 2000. Bud regeneration from inflorescence explants for rapid propagation of geophytes in vitro. Plant Cell Rpt. 19:845-850.