Please use this identifier to cite or link to this item: http://hdl.handle.net/11455/29199
標題: 無子葡萄選育技術之研究
Studies on Techniques for Breeding Seedless Grapes
作者: 許怡萱
Hsu, Yi-Hsuan
關鍵字: seedlessness;無子;triploid;embryo rescue;endosperm;marker assisted selection;三倍體;胚拯救;胚乳;標誌輔助選拔
出版社: 園藝學系所
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摘要: 
本研究的目的之一為測試二倍體`貝利A´與四倍體`巨峰´葡萄雜交後胚珠進行離體培養之適當取胚時間及培養條件以獲得三倍體植株。結果顯示,雜交授粉後之最佳取胚時間為花後49-56天,可獲得較高之胚珠萌芽率。將胚珠先常溫26℃及光周期16/8 (明/暗)小時培養一段時間後,經低溫(6 ± 1℃)暗處理2個月,再移回常溫26℃及光周期16/8 (明/暗)小時培養可促進胚珠萌芽率。
本研究另測試`貝利A´及`巨峰´葡萄胚乳培養之最適合誘導癒傷組織之胚乳發育階段及後續繼代之培養條件。結果發現,取花後2週內胚乳誘導癒傷組織率較高;MSMO基本培養基添加500ppm酪蛋白水解物誘導癒傷組織產生率較高;胚乳留胚誘導癒傷組織較佳。癒傷組織繼代培養於TDZ、BA或是TDZ + NAA及BA + NAA培養基中皆無法誘導不定芽產生。`貝利A´胚乳癒傷組織培養在NOA及BA培養基中癒傷組織生長效果較差,且無法分化成體胚。`巨峰´胚乳癒傷組織以常溫26℃及光周期16/8 (明/暗)小時培養於10 µM NOA + 10 µM BA培養基中約5個月後有多細胞團產生,移至含2,4-D及BA不同濃度組合之培養基暗培養3個月後,可以在顆粒狀癒傷組織裡找到較多的多細胞團。
本研究另一主要目的為尋找與`巨峰´無子變異株之無子性狀連結之分子標誌,以做為未來相關四倍體無子葡萄育種時之選拔工具。利用RAPD分析比較`巨峰´與`巨峰´無子變異株,經篩選後發現M02、W14及AA16三支引子均於`巨峰´葡萄產生`巨峰´無子變異株沒有之特有條帶,將來可用來區分此二品種。

The ovules harvest time and culture conditions for in vitro culture of embryo from crosses between diploid `Muscat Bailey A´ and tetraploid `Khoyo´ grapes were examined. The results indicated that the optimal ovules harvest time were 49-56 days after blooming which had higher percentage of ovule germination. Ovules cultured at 26℃ for a period of time, followed by incubated at 6 ± 1℃ in the dark for two months, and then transferred back to 26℃ with photoperiod of 16/8 (light/dark) hours promoted ovule germination rates.
This study also tested the effect of developmental stage of endosperm and subculture conditions for callus initiation from endosperms of `Muscat Bailey A´ and `Khoyo´ grapes. Higher callus induction rates were obtained from endosperms of 2 weeks after blooming from both `Muscat Bailey A´ and `Khoyo´ grapes. Endosperms cultured with embryo on MSMO medium supplemented with 500ppm casein hydrolysate promoted callus induction. The endosperm callus subcultured on medium cataining TDZ or BA alone, or in combination with NAA could not induce adventitious shoots. The endosperm callus of `Muscat Bailey A´ grew poorly on the subculture medium supplemented with NOA and BA and could not induce somatic embryogenesis. However, the endosperm callus of `Khoyo´ grapes produced multicellular mass on the subculture medium containing 10 µM NOA and 10 µM BA after five months of incubation at 26℃ with photoperiod of 16/8 (light/dark) hours. More multicellular masses were found in granular endosperm callus which were incubated in the dark for three months on media supplemented with different concentration of 2,4-D and BA.
Another important objective of this study was to find molecular markers linked to the seedlessness character of `Khoyo´ mutant which might be used as a selection tool in breeding of tetraploid seedless grapes in the future. Using RAPD analysis, polymorphic bands between `Khoyo´ and `Khoyo´ mutant were found by primers M02, W14 and AA16. Those bands were present in the `Kyoho´ and absent in the `Kyoho´ mutant which could be used to distinguish the two cultivars.
URI: http://hdl.handle.net/11455/29199
其他識別: U0005-2308201010415300
Appears in Collections:園藝學系

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