補血草屬植物之低春化需求選育與指標、開花度積值及親緣關係

Breeding of Low Vernalization-requiring Cultivars/Lines, Selection Indices, Thermal Time of Flowering, and Phylogenetic Relationship in Limonium

張元聰(Yuan-Tsung Chang)

指導教授 ： 楊雯如

共同指導教授 ： 葉德銘(Der-Ming Yeh)

國立臺灣大學/生物資源暨農學院/園藝暨景觀學系/博士(2014年)

https://doi.org/10.6342/NTU.2014.01086

摘要

本論文旨在選育低春化需求之補血草屬(Limonium)品種/系，使其更適合在臺灣平地栽培，採用耐熱性指標比較育成之早花品種和商業品種之差異，以度積值模式建立開花之溫度條件，並探討臺灣不同產地原生種之歧異度。
採用星辰花[L. sinuatum (L.) Mill.]‘超群’、‘福神’及‘達摩’等混色系列，以標定早花植株收取種子，將早花植株混合選種，並持續選拔四至五代後，其花期均較原始族群提早70-80天左右，顯示早花選種的育種策略有效。所選得之‘臺南1號’及商業品種‘百萬紫’以組織培養繁殖，進行開花比較試驗，‘臺南1號’經35/30℃處理4週後，定植於塑膠布溫室之到花天數為82.7天，‘百萬紫’為102.7天，顯示‘臺南1號’之開花需冷性較低，因而早花。選育之‘臺南1號’是針對亞熱帶氣候條件下選育的品種，花萼紫色，可在臺灣平地10月定植，1月自然開花，能在價格最好的冬季採收切花供應市場需求。
對栽培於35/30及20/15℃下的‘臺南1號’及‘百萬紫’植株調查葉片細胞膜熱相對傷害值(relative injury, RI)、丙二醛(malondialdehyde, MDA)濃度及葉綠素螢光值(Fv/Fm、Fo)變化，結果顯示兩品種於35/30℃處理之RI值較20/15℃處理者低。在35/30℃處理6-12天內，‘臺南1號’之RI值持續下降，MDA濃度持續上升至約64-86 nmol•g-1，12天後之RI值及MDA濃度皆維持穩定不再變動。而‘百萬紫’在35/30℃處理6-15天內，MDA濃度持續上升至約154 nmol•g-1，之後RI值才穩定降低。兩品種Fv/Fm及Fo值之表現趨勢類似，品種間以‘百萬紫’ Fo值明顯較高，在35/30℃處理之Fv/Fm值下降及Fo值上升幅度皆以‘百萬紫’較‘臺南1號’為高。以選育之早花品系和商業品種為材料，調查定植前處理35/30℃當日及第18天時之上述耐熱性生理指標，以第18天與處理當日之測量值比值與定植至開花之日數進行廻歸分析，較晚開花的品種/系有較高之RI值比值、MDA濃度比值及Fo值比值，與較低之Fv/Fm值比值。顯示在高溫下測量葉片RI值、丙二醛濃度及葉綠素螢光值，可檢測星辰花品種之耐熱性與開花時間。
建立溫度對兩星辰花品種花芽創始及發育之基礎溫度(base temperature, Tb)、最適溫度(optimum temperature, Topt)及最大溫度(maximum temperature, Tmax)，並以此三基點建立度積值(thermal time, ℃d)。取具10-15片葉之植株置於3-21℃分別處理14-28天，再移至24℃生長箱環境後，調查花下葉片數及到花芽可見天數。以花下葉片數之倒數與處理溫度進行迴歸分析，結果顯示‘臺南1號’之Tb為-3.0℃，Topt為12.3℃，Tmax為22.1℃，而‘百萬紫’之Tb為-5.2℃，Topt為11.3℃，Tmax為24.7℃。以到花芽可見天數之倒數與處理溫度進行迴歸分析，結果顯示‘臺南1號’之Tb為-1.0℃，Topt為12.0℃，Tmax為27.2℃，而‘百萬紫’之Tb為-0.9℃，Topt為10.9℃，Tmax為23.4℃。‘臺南1號’之花芽創始或到可見花芽之速率約在230℃d時達到飽和春化反應，而‘百萬紫’在給予之春化處理範圍內尚未飽和，顯示‘臺南1號’春化所需度積值較‘百萬紫’少。將花芽已創始之星辰花植株置於15-36℃下，並調查其花芽發育階段。取創始至盛花(第一花穗花朵全數開放)天數之倒數，計算其花芽發育速率，結果顯示‘臺南1號’花芽發育Tb為10.7℃，而‘百萬紫’為10.1℃，而‘臺南1號’從花芽創始至盛花之度積值為373.8℃d，‘百萬紫’則為407.2℃d，顯示‘臺南1號’花芽發育速度較‘百萬紫’稍快。所建立模式未來可應用在實際栽培生產時，控制環境溫度以調節春化種苗生產或切花出貨時間。
為選育早花水晶花(L. hybrids)品種/系以適合亞熱帶地區栽培，將其與臺灣原生且能週年開花之石蓯蓉[L. sinense (Girard) Kuntze]雜交，選拔開花品質優良之早花後代和商業品種進行性狀與開花之比較試驗。將具10-15片葉之‘臺南2號’及‘黃鑽’組培苗於定植前在15/10℃處理4週後定植於溫室，結果以‘臺南2號’較‘黃鑽’早10天開花，花莖亦較長。而定植前35/30℃處理會使‘臺南2號’到開花時間較15/10℃處理者晚約10天，然‘黃鑽’經高溫處理者至定植150天時仍未開花。選育品種‘臺南2號’花萼黃色且到開花時間早，適合在亞熱帶氣候條件下生產切花，臺灣平地可在10月種植而於冬季價格最好的時候開花，供應市場需求。
收集13個不同原生地石蓯蓉族群及4個不同原生地烏芙蓉[L. wrightii (Hance) Kuntze]族群之種子，於同一環境下播種栽培後調查分析。以ISSR分子標誌分析結果顯示，共有16條引子、220個條帶具有多形性，臺灣地區原生石蓯蓉可分為四群，分別為馬祖及金門一群，屏東龍坑獨自一群，臺北及基隆嶼為第三群，第四群為本島西岸及鄰近外島族群。外表性狀差異和地理位置之相關性較差，顯示以ISSR分析之結果較符合地理位置之遠近。原生於屏東佳樂水及南仁港之烏芙蓉雖花色或外形有些微差異，但外表性狀及ISSR分析結果顯示為同一族群，而原生於蘭嶼者花色及形態和原生本島者具有差異，原生於綠島者外形較接近蘭嶼族群，但具有多種花色。

關鍵字

早花選種 ； 春化作用 ； 丙二醛 ； 光系統II最大光子利用效率 ； 溫度三基點 ； 度積值 ； 簡單序列間重複 ； 分子分類法

並列摘要

The objectives of this study were to select Limonium cultivars/lines with low vernalization requirement, which are suitable for lowland production in Taiwan. Heat-tolerance-related physiological indices of early-flowering selected Limonium cultivars/lines were compared with commercial cultivars, and cardinal temperatures and thermal times model of flower initiation and development were established. Genetic diversity of Taiwan-native Limonium species from different habitats was also analyzed.
Plants of L. sinuatum ‘Excellent Mix’, ‘Turbo Mix’, and ‘Fortress Mix’ were used for selection. The early-flowering individual plants were labelled for mass selection. After selection for 4 to 5 generations, the selected plants flowered 70 to 80 days earlier than the parent populations. The selected ‘Tainan No.1’ and the commercial cultivar Million Purple were mass-propagated by tissue culture. After treated at pre-planting temperature of 35/30℃ for 4 weeks, the average time from planting to flowering of ‘Tainan No.1’ and ‘Million Purple’ were 82.7 and 102.7 days, respectively. Results showed that ‘Tainan No.1’ had lower low temperature requirement and subsequently flowered earlier. ‘Tainan No.1’ has purple calyx and is selected for cut flower production in sub-tropical climates. In the lowland area of Taiwan, ‘Tainan No. 1’ can be planted in October and flowers naturally starting from December with peaked flowering in January and February when the price is high.
Plants of ‘Tainan No.1’ and ‘Million Purple’ were grown under day/night temperature of 35/30℃, and measured for leaf relative injury (RI), malondialdehyde (MDA) concentration, and chlorophyll fluorescence. For both cultivars, leaf RI values were lower in plants at 35/30℃ than at 20/15℃. Leaf RI decreased and MDA concentration increased to 64-86 nmol•g-1 in ‘Tainan No.1’ at 35/30℃ during 6 to 12 days, and both leaf RI and MDA remained unchanged thereafter. In contrast, leaf MDA increased from 59 to 154 nmol•g-1 in ‘Million Purple’ when treated at 35/30℃ from 6 to 15 days, and leaf RI decreased gradually afterwards. Changes of Fv/Fm and Fo value were similar in both cultivars. ‘Million Purple’ had higher Fo values than ‘Tainan No.1’. ‘Million Purple’ exhibited more decreased Fv/Fm and increased Fo than ‘Tainan No.1’. Late flowering cultivars/lines were those with higher ratios (Day 18 vs Day 0) of leaf RI value, MDA concentration, and Fo value, and lower ratio of Fv/Fm value.
The base (Tb), optimum (Topt), and maximum (Tmax) temperatures were determined and the thermal times required for flower initiation and development were calculated in Limonium cultivars ‘Tainan No.1’ and ‘Million Purple’. Plants with 10 to 15 leaves were treated under 3 to 21℃ for 14 to 28 days, and these plants were then placed to 24℃growth room conditions for further growth. Leaf number below the flower and days to visible flower bud were recorded. Rate of progress to flower initiation was calculated as the reciprocal of leaf number below the flower. The estimated Tb, Topt, and Tmax were -3.0, 12.3, and 22.1℃ in ‘Tainan No.1’, and -5.2, 11.3, and 24.7℃ in ‘Million Purple’. Rate of progress to flower bud visibility was calculated as reciprocal of days to visible flower bud. The estimated Tb, Topt, and Tmax were -1.0, 12.0, and 27.2℃ in ‘Tainan No.1’, and -0.9, 10.9, and 23.4℃ in ‘Million Purple’. ‘Tainan No.1’ had saturated vernalization response with vernalizing thermal time of ca. 230℃d, whereas ‘Million Purple’ did not reach saturated response under the conditions constructed. Post-initiation development was studied in plants placed under 15 to 36℃, and floral developmental stage was recorded. The estimated Tb for flower development were 10.7 and 10.1℃ in ‘Tainan No.1’ and ‘Million Purple’, respectively. The thermal times required from flower initiation to anthesis were estimated as 373.8℃d in ‘Tainan No.1’, and 407.2℃d in ‘Million Purple’.
High vernalization-requiring ‘Yellow Diamond’ was crossed with L. sinense (Girard) Kuntze, a Taiwan-native species, with year-round flowering characteristics. Tissue cultured plants of selected early-flowering ‘Tainan No.2’ and ‘Yellow Diamond’ were used. Plants with 10-15 leaves were transferred to growth chambers at pre-planting day/night temperatures of 15/10 and 35/30℃ for 4 weeks and then planted in a plastic house. Plants of ‘Tainan No.2’ flowered 10 days earlier and had longer inflorescences than ‘Yellow Diamond’ for the 15/10℃ pre-planting treatment. Flowering of ‘Tainan No.2’ at 35/30℃ was delayed by 10 days compared with those at 15/10℃. In contrast, ‘Yellow Diamond’ plants did not flower after 150 days of cultivation after the 35/30℃ treatment. ‘Tainan No.2’ has yellow calyx and is selected for cut flower production in subtropical climates. In the lowland area of Taiwan, ‘Tainan No.2’ can be planted in October and flowers naturally in winter when the price is high.
Seeds of L. sinense from 13 habitats and L. wrightii (Hance) Kuntze from 4 habitats were collected and grown in Tainan under the same conditions for investigation. Results showed that 16 ISSR primers and 220 induced bands were polymorphic. Limonium sinense accessions could be divided into four groups. Group 1 included those from Jinmen and Mazu, and those from Longkeng, Bingdong forms a single group. Group 3 included those from Taipei and Jilong, and Group 4 included those from western seashore and near islands of Taiwan. Relationship between morphological traits and originate habitats was poor. Results showed that ISSR marker could be used for distinguishing Taiwan-native L. sinense plants from different habitat. Limonium wrightii derived from Nanrengang and Jialeshui, Bingdong were not different based on morphological traits and ISSR marker, despite of slight difference in flower color and other morphologies. Plants derived from Lanyu island, Taidong were different with those from Taiwan island in flower color and other morphologies. Plants derived from Ludao, Taidong were similar to those from Lanyu, but had more flower color variation.

並列關鍵字

early-flowering selection ； vernalization ； malondialdehyde ； maximum quantum efficiency of photosystem II ； cardinal temperatures ； thermal time ； inter-sequence simple repeat ； molecular taxonomy

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