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研究生:陳柏吟
研究生(外文):Bo-Yin Chen
論文名稱:香石竹之種間雜交及石竹多倍體化
論文名稱(外文):The Interspecific Hybridization of Carnation and the Polyploidization of Dianthus.
指導教授:陳彥銘陳彥銘引用關係
口試委員:潘怡君李嘉雯黃建誌
口試日期:2018-07-16
學位類別:碩士
校院名稱:國立中興大學
系所名稱:園藝學系所
學門:農業科學學門
學類:園藝學類
論文種類:學術論文
論文出版年:2019
畢業學年度:107
語文別:中文
論文頁數:133
中文關鍵詞:香石竹種間雜交多倍體誘變恢復稔性
外文關鍵詞:carnationinterspecific hybridizationpolyploid mutantrestore fertility.
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本論文利用石竹屬物種進行花粉活力測定、胚珠數量調查及使用收集之香石竹品種進行品種特性調查,如到花日數、單花壽命、瓶插壽命及吸水量之評估,以作為後續育種親本之選擇。試驗使用香石竹商業品種種子系-盆花ʻLillipot’、種子系-切花ʻJabo’選拔之單株、營養系Standard type ʻOptimistic’及Spray type ʻFestival’切花與原生石竹屬物種Dianthus palinensis、石竹屬商業品種D. japonicus ʻPurple Plum’與D. barbatus ʻLord Breanthus’、石竹種間雜交種D. hybrida ʻLS’作為親本進行正反雜交試驗與雜交後代性狀之調查。並利用流式細胞儀與RAPD 技術驗證雜交後代。此外利用秋水仙素進行石竹種間雜交種ʻ巴陵紫雲’多倍體誘變,獲得恢復稔性之單株,以進行後續育種工作。
香石竹種子系品種花粉活力較營養系品種花粉活力高,石竹屬物種花粉活力亦具有較香石竹高的花粉活力。種子系品種到花日數短於營養系品種。種子系品種單花壽命介於5.4-7.6天,營養系品種瓶插壽命介於7.0-14.3天。
香石竹與石竹屬物種進行雜交,以ʻFestival’與石竹屬物種較佳的雜交親和性。而香石竹ʻFestival’與石竹屬物種之雜交親和性排序,依序為D. palinensis、D. barbatus ʻLord Breanthus’、D. japonicus ʻPurple Plum’及D. hybrida ʻLS’。此外,香石竹ʻFestival’與石竹屬物種進行雜交,分別於D. japonicus ʻPurple Plum’為父本,D. palinensis與D. hybrida ʻLS’作為母本時,無法獲得雜交後代,顯示有單向不親和現象之發生。
種間雜交後代植物性狀多數特徵具超越雙親或呈現中間型性狀。如香石竹ʻFestival’作為母本與D. palinensis進行雜交,其雜交後代皆具有花香味。ʻFestival’作為母本與D. japonicus ʻPurple Plum’進行雜交,其雜交後之葉蠟層與父本一樣具有革質葉片。香石竹ʻFestival’與D. barbatus ʻLord Breanthus’之種間雜交組合,不論正交或反交的雜交後代,花瓣皆為重瓣。ʻFestival’作為母本與D. hybrida ʻLS’進行雜交,其雜交後代與雙親一樣具有花香味。雜交後代利用流式細胞儀檢測其相對DNA含量,皆位於雙親之間。利用RAPD 驗證雜交後代,皆具雙親之特異性條帶。此外,亦成功選育具早生、重瓣、單花壽命長之單株。
獲得之種間雜交後代D. hybrida ʻLO×P-1’及D. hybrida ʻLO×LS-1’與其親本進行回交試驗,及與商業品種之香石竹ʻOptimistic’、ʻFestival’及美國石竹D. barbatus ʻLord Breanthus’進行再雜交,成功選育花瓣數增加、單花壽命延長、新特殊型態,具商業價值潛力之後代。
石竹種間雜交種ʻ巴陵紫雲’以不同濃度秋水仙素處理,共獲得54株多倍體誘變株。此外經秋水仙素處理後,混倍體誘變株其花藥可正常釋放花粉,成功獲得恢復稔性之誘變植株,花粉活力可達8.53%。
The present research, the Dianthus species is employed to determine pollen germination, investigate the ovule quantity and use collected cultivars of carnation to look into characteristics, such as evaluations of days to flowering, longevity of flower, vase life, and water absorption as an assessment for subsequent breeding parents. Utilize seed line of commercial cultivars of potted carnation ʻLillipot’ and single plant selected from seed line of commercial cultivars of cut carnation ʻJabo’; clone of standard type ʻOptimistic’ and spray type ʻFestival’ of carnation, as well as native species Dianthus palinensis, commercial cultivars of D. japonicus ʻPurple Plum’, D. barbatus ʻLord Breanthus’ and D. hybrida ʻLS’ in Dianthus as parents to conduct reciprocal crosses and investigate the characters of hybrids in the test. In addition to using flow cytometer and RAPD technology to verify hybrids. Moreover, inducing polyploidization of the Dianthus interspecific hybrid ʻPalin Purple Cloud’, and the fertility mutants recovered for subsequent breeding.

The carnation seed varieties pollen viability is higher than that of the clone strains, while the Dianthus species pollen viability is higher than carnation. The days to flowering of seed varieties are shorter than those of clonal varieties. The flower life span of the seed varieties is between 5.4 and 7.6 days, while the vase life of the clonal varieties is between 7.0-14.3 days.

When carnation and the Dianthus species interbreed, ʻFestival’ and the Dianthus species display better cross-compatibility. The cross-compatibility of D. caryophyllus ʻFestivalʻ and the Dianthus species are sequenced as follows: D. palinensis, D. barbatus ʻLord Breanthus’, D. japonicus ʻPurple Plum’ and D. hybrida ʻLS’, In addition, when D. caryophyllus ʻFestival’ and the Dianthus species interbreed where D. japonicus ʻPurple Plum’ acts as a male parent and D. palinensis and D. hybrida ʻLS’ as a female parent, a filial generation cannot be obtained, which shows the phenomenon of one-way incompatibility.

The inter-species hybridization generation plants are characterized by the traits beyond the parents or intermediate characters. For example, when D. caryophyllus ʻFestival’ acting as a female parent cross-breeds with D. palinensis, their hybrids have floral fragrances. When ʻFestival’ acting as a female parent is crossed with D. japonicus ʻPurple Plum’, the leaf wax layer after hybridization has the same leathery leaves as the male parent. For the interspecies cross combination between D. caryophyllus ʻFestival’ and D. barbatus ʻLord Breanthus’, the petals are double, regardless of orthogonal or reciprocal hybrid offspring. When ʻFestival’ acting as a female parent cross-breeds with D. hybrida ʻLS’, their hybrid offspring have the same floral scent as their parents. A flow cytometry is adopted to detect the relative DNA content of hybrid offspring, which is between the male and female parents. RAPD is used to validate the filial generation, finding it has parents’ specific bands. Furthermore, a single plant with an early variety, double flowers, and long flower natural life is successfully bred.

The acquired inter-species hybridization generations ʻLO×P-1’ and ʻLO×LS-1’ undergo crossing back experiments with their parents, and then are re-crossed with commercial varieties D. caryophyllus ʻOptimistic’, ʻFestival’ and D. barbatus ʻLord Breanthus’ before successfully breeding the descendants with increased petal counts, prolonged flower life-span, new special types, and commercial value and potential.

The interspecific hybrid Dianthus ʻPalin Purple Cloud’ is treated with varying concentrations of colchicine, with a total of 54 mutagenic polyploid plants acquired. Moreover, following colchicine treatment, the anther of the mutagenic mixoploid strains can release pollen normally and obtain successfully the mutagenic plant that restores fertility, with pollen viability reaching 8.53%.
目次
摘要……………………………………………………………………………...…i
Abstract………………………………………………………….……………....…ii
目次…………………………………………..………………….…..…………... iv
表目次……………………………………………………………….…………….v
圖目次……………………………………………………………….…………..viii
前言(Introduction)……………………………...………………………………1
前人研究(Literature review)………………………...…………………………2
一、石竹科分類及石竹屬分布………………………………...........………2
二、石竹屬重要物種……………………………………………………..…...3
三、石竹屬育種方式…………………………………………………..……...5
四、香石竹栽培與育種史…………………………………………..………...7
五、香石竹育種目標…….……………………………………..….………...10
材料與方法(Materials and methods)……………………………….………….14
一、試驗材料繁殖及管理………………………………………………..….14
二、試驗方法…………………………………………………………..…….14
(一) 石竹屬物種特性調查…………………………...………………..14
(二) 香石竹品種特性調查…………………………...………………....15
(三) 種間雜交…………………………...………………....…………..16
(四) 多倍體誘變…………………………...………………....………..16
(五) 植物性狀調查…………………………...…………….…...……..17
(六) 相對DNA含量測定…………………………...……….………..17
(七) RAPD分析…………………………...………………...………...17
(八) 統計分析…………………………...………………...…..……….18
結果(Result)………….………………………………………………………...19
一、花粉活力測定....................……………………………………..…….....19
二、香石竹品種特性調查……………………………………………..…….20
三、石竹屬物種與香石竹商業品種雜交試驗………………………..…….22
四、雜交後代植物性狀調查………………………………………….……..25
五、種間雜交後代DNA含量檢測與RAPD技術分析……….………..….36
六、種間雜交種ʻ巴陵紫雲’之誘變處理……………………...……….….37
討論(Discussion)………………………………………………………………39
一、花粉活力檢測……………………………………..…………………….39
二、石竹屬種間雜交、回交與三交…………………………………..…….41
三、石竹屬植物雜交後代性狀表現……………….……...……….………..46
四、多倍體誘變…………………………………..……………..……………..49
參考文獻(Reference)………………………………………………………….125
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