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研究生:吳佩眞
研究生(外文):Pei-Jhen Wu
論文名稱:長豇豆之生殖發育、花藥開裂時間及花粉活力
論文名稱(外文):The Reproductive Development, Anther Dehiscence and Pollen Viability of Yard-long Bean
指導教授:曹幸之曹幸之引用關係
口試委員:羅筱鳳張有明
口試日期:2011-01-13
學位類別:碩士
校院名稱:國立臺灣大學
系所名稱:園藝學研究所
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:2011
畢業學年度:99
語文別:中文
論文頁數:76
中文關鍵詞:三苯基氯化四氮唑雙醋黃螢光苯胺藍
外文關鍵詞:2,3,5-triphenyl tetrazolium chloridefluorescein diacetateaniline blue
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長豇豆 (Vigna unguiculata ssp. sesquipedalis (L.) Verdc) 性喜溫、耐旱又耐濕,對環境適應範圍廣,是臺灣重要夏季蔬菜之一。但栽培上常有種傳病毒病為害,以及近年來環境變遷導致產量、品質受影響。長豇豆為自花授粉作物,雜交育種有授粉效率低、結籽數少的限制。有關於長豇豆之生殖特性,包括開花、花藥開裂時間及花粉壽命與活力的報告甚少。本研究以‘三尺青皮’與‘麗人’兩蔓性品種,以及‘矮長豆’與‘農友矮生’兩矮性品種為試驗材料,分別栽種於台灣大學試驗田區及人工氣候室日/夜溫30/25 oC與25/20 oC條件。植株進入抽蔓期所需之時間以25/20 oC處理最久,‘三尺青皮’、‘麗人’分別需要22.6與21.6日 (自播種算起)。植株在25/20 oC 處理也最慢進入生殖生長;但各品種第一花苞出現之節位在三個環境下,均無明顯差異。其中‘三尺青皮’花苞出現節位最高 (8.3 – 8.7節),其他三個品種花苞出現節位平均為第3.0 - 3.7節。各品種於進入生殖生長期後分成30/25 oC、25/20 oC兩處理,分別調查其花藥開裂時間,自花開前一天7:00 pm起每整點取樣觀察,並與田間結果相比較。供試長豇豆於花開前一天7:00 - 8:00 pm開始花藥開裂,於凌晨零時全數開裂完畢。以Alexander、TTC (2, 3 ,5 - triphenyl tetrazolium chloride) 及FDA (fluorescein diacetate) 三種染色方法評估花粉活力,前兩種染色法之染色值較高,FDA檢測之染色率偏低 (< 30%)、較不易用以評估花粉活力。以TTC染色法較能反映出長豇豆花粉活力隨時間之變化;栽培於人候室之植株花粉可染率、由花開前一日晚上8:00之96.98% 隨時間遞降,至花開當日4:00 pm降至53.29%。於花開前一日4:00 pm去雄並進行品種內之授粉,在25/20 oC環境下‘三尺青皮’與‘矮長豆’分別有70% 及15% 的成功率,顯示長豇豆柱頭在花開前一日傍晚,已具可授性。此時花藥尚未開裂,人工除雄及授粉作業可於此時間一併進行。以B&K培養基為基礎配方,蔗糖濃度50% 才能維持長豇豆花粉不破裂,但花粉不能萌發。培養基添加其他物質,如:聚乙二醇 (polyethylene glycol)、小牛血清蛋白 (bovine serum albumin)、激勃素 (gibberellin acid) 或長豇豆柱頭組織、調整pH值或其他鹽類濃度與種類,改變培養基型態及培養方式都無法使長豇豆花粉管伸長超過花粉直徑,顯示長豇豆花粉屬於難離體發芽的種類。

Yard-long bean (Vigna unguiculata ssp.sesquipedalis (L.) Verdc.) thrives in warm weather with good tolerance to drought and to moisture. It has a wide range of soil adaptation and is an important summer vegetable in Taiwan. However, the crop succumbs to seed borne viral diseases and abrupt environmental changes often result in reduced pod yield and quality. Being a self-pollinated legume, yard-long bean usually has low seed set upon manual crossing. There is few report on the reproductive biology of the yard-long bean and it is essential to know the flowering and the pollen behavior of the plant in order to ensure the success in commercial production and the seed set in breeding work. Four yard-long bean cultivars, including two viny (cvs. ‘Green-pod Kaohsiung’ and ‘Milady’) and two dwarf types (cvs. ‘Bush’ and ‘K.Y. Bush’) were grown in the open field of experimental farm and the phytotron with day and night temperature regimes of 30/25 oC and 25/20 oC at National Taiwan University. It took 22.6 and 21.6 days after sowing for cvs. ‘Green-pod Kaohsiung’ and ‘Milady’, respectively to start vining under 25/20 oC. Plants grown in the field and 30/25 oC of phytotrons were earlier to vine and to develop reproductive structures than plants under 25/20 oC, however, the node position of the first flower bud remained same in three environments, i.e. node 8.3 - 8.7 for ‘Green-pod Kaohsiung’ and node 3.0 - 3.7 for the other three varieties. All plants were grown in 25/20 oC till visible inflorescence shown and then they were either moved to 30/25 oC or kept at 25/20 oC. Flower buds due to bloom or open the next day were collected at 7:00 pm and thereafter at 4-hr intervals to examine the timing of anther dehiscence. The results showed that flowers of yard-long beans completed the anther dehiscence at 0:00 am of the day of anthesis. Pollen viability was tested by three staining methods. Both Alexander and TTC (2,3,5-triphenyl tetrazolium chloride) methods gave higher staining rate than FDA (fluorescein diacetate) test which showed less than 30% of staining. TTC staining results may indicate the temporal change in pollen viability, the staining rate was 96.98% from pollens collected at 8 pm of the day before anthesis and it decreased to 53.29% at 4:00 pm of day of anthesis. The pod set rates were 70% and 15% for cvs. ‘Green-pod Kaohsiung’ and ‘Bush’, respectively from pollinated buds due to bloom the next day under temperature of 25/20 oC. To make a hybridization cross, both emasculation and pollination can be carried out for yard-long bean in the afternoon of day before anthesis. All pollens failed to germinate in vitro on B&K or other modified culture medium tested. Sucrose as high as 50% was needed to prevent pollen from burst, supplements such as polyethylene glycol, bovine serum albumin, GA and crushed yard-long bean stigma were added with no success. Other modification included pH and formulations, no pollen germinated in excess of its diameter. Yard-long bean pollen is difficult species to germinate in vitro.

口試委員會審定書...............................i
致謝..................................................ii
中文摘要............................................iii
英文摘要...........................................iv
目錄..................................................v
圖表目錄...........................................vi
第一章、前言.....................................1
第二章、前人研究..............................3
  一、豇豆植物特性與溫度影響.......3
  二、花藥開裂與花粉釋出..............6
  三、花粉活力..............................9
第三章、材料與方法.........................20
  一、試驗材料............................20
  二、栽培環境與植株生長............20
  三、花藥開裂與花粉萌發............21
  四、花粉活力檢測......................22
第四章、結果...................................30
  一、環境對長豇豆生長之影響......30
  二、長豇豆花藥開裂時間............31
  三、花粉活力檢測......................32
第五章、討論...................................59
結論................................................65
參考文獻.........................................66
附錄................................................74


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