臺灣博碩士論文加值系統

本研究以‘台農17號’與‘台農20號’鳳梨為材料，以不同濃度NAA與BA組合添加於培養基中，誘導不同培植體生成癒傷組織，以及體胚誘導與器官發生，並測試EMS誘變與羥脯胺酸篩選之適合處理條件，另外，檢測EMS誘變品系對低溫逆境之耐受性及脯胺酸含量。
‘台農17號’鳳梨以葉片白色基部進行癒傷組織誘導，以1ppmNAA+8ppmBA較佳。 ‘台農20號’以葉片白色基部進行誘導，則以在2ppm NAA+8ppm BA之處理具有最高誘導率。
EMS誘變條件與羥脯胺酸篩選條件之試驗結果顯示，‘台農17號’鳳梨組培瓶苗以0.4% EMS 24小時為誘變條件，處理後35天存活率為62.5%，而羥脯胺酸18mM培養28天之存活率為47.5%。‘台農20號’鳳梨組培瓶苗則以0.4% EMS 24小時為誘變條件，於處理後35天有62.5%的存活率，羥脯胺酸18mM培養28天存活率只有為27.5%。因此，‘台農17號與‘台農20號’ 鳳梨對EMS之耐性相近，但‘台農20號’對羥脯胺酸之耐性則低於‘台農17號’鳳梨。
‘台農17號’鳳梨EMS誘變品系TN17M1-36-2及‘台農20號’EMS誘變品系TN20M1-4與TN20M1-6之第二子代(P2)，均較未誘變系顯著含有較高脯胺酸含量，而在經由低溫(5ºC，14天)處理後，前述誘變系脯胺酸含量顯著上升，且也有較高之存活率，結果顯示，EMS誘變可產生較耐逆境之後代，且羥脯胺酸可用於鳳梨誘變植株耐逆境後代之篩選。

Using ‘Tainung No.17’ and ‘Tainung No.20’ pineapple (Ananas comosus (L.) Merrill.) as plant material, different combinations of NAA and BA were tested for callus induction, somatic embryo induction and organogenesis. The optimal treatment conditions for EMS mutagenesis and hydroxyproline selection were tested. In addition, cold tolerance and proline contents of EMS induced mutants were also examined.
In callus induction and proliferation, white basal part of ‘Tainung No.17’ pineapple leaves cultured in 1ppm NAA+ 8ppm BA had a higher induction rate. The highest induction rate occurred in the treatment of 2ppm NAA + 8ppm BA for ‘Tainung No.20’ pineapple.
The results of EMS mutagenesis treatment and hydroxyproline selection condition for ‘Tainung No.17’ tissue culture plantlets indicated that the survival rate was 62.5% 35days after 0.4% EMS 24 hours treatment and 47.5% after 28days of 18mM hydroxyproline treatment. For ‘Tainung No.20’ pineapple, the survival rate was 62.5% in the treatment of 0.4% EMS 24 hours, but only 27.5%sfter 28days of 18mM hydroxyproline treatment. As a result, the tolerance for ENS was similar for ‘Tainung No.17’ and ‘Tainung No.20’ pineapples, but the tolerance for hydroxyl- proline was lower for ‘Tainung No.20’ than for ‘ Tainung No.17’ pineapple.
The second generation progenies (P2) of ‘Tainung No.17’ pineapple EMS mutant TN17M1-36-2 and ‘Tainung No.20’ pineapple EMS mutants TN20M1-4 and TN20M1-6 had higher proline contents than the control plantlets. The proline contents of the above mutants were significantly increased after cold treatment (5ºC, 14 days) and had higher survival rates than the control. The results showed that EMS treatment could generate stress tolerant progenies and hydroxyproline could be used for stress tolerant progeny selection.

目次
摘要 i
Abstract ii
目次 iii
表目次 v
圖目次 vi
一、前言 1
二、前人研究 2
(一)鳳梨組織培養 2
1.果冠休眠腋芽增殖苗 2
2.癒傷組織植體再生 2
3.體胚的培養與增殖 3
(二)EMS(ethyl methanesulfonate)誘變 3
1.烷類誘變劑誘變機制 3
2.EMS誘變機制 3
3.EMS在作物育種上之應用 4
4.化學性誘變育種在鳳梨上之應用 5
(三)誘變後代之選拔 5
1.逆境型環境選拔 5
2.植物耐逆境性狀選拔 6
(四)Hydroxyproline 在作物育種上之應用 6
1.Hydroxyproline型態與形成機制 6
2.脯胺酸於植物逆境生理之意義 7
3.羥脯胺酸對植物生理之意義 7
4.羥脯胺酸與脯胺酸在作物育種上之應用 7
三、材料與方法 9
試驗一、‘台農17號’與‘台農20號’鳳梨不同部位之癒傷組織誘導 9
試驗二、‘台農17號’與‘台農20號’鳳梨癒傷組織之增殖 10
試驗三、EMS誘變‘台農17號’及‘台農20號’鳳梨組織培養小苗之最適條件 10
(一)EMS 誘變‘台農17號’鳳梨組織培養小苗之最適條件 10
(二)EMS 誘變‘台農20號’鳳梨組織培養小苗之最適條件 10
試驗四、EMS誘變‘台農20號’癒傷組織之增殖 11
試驗五、羥脯胺酸篩選‘台農17號’及‘台農20號’鳳梨組培小苗之最適條件 11
試驗六、低溫處理‘台農17號’及‘台農20號’鳳梨經EMS誘變及羥脯胺篩選後之組培小苗 12
(一)低溫處理‘台農17號’及’台農20號’組培小苗 12
(二)‘台農17號’鳳梨組培小苗 12
(三)‘台農20號’鳳梨組培小苗 13
四、結果 14
試驗一、‘台農17號’及‘台農20號’鳳梨不同部位之癒傷組織誘導 14
試驗二、‘台農17號’及’台農20號’鳳梨癒傷組織之增殖 22
試驗三、EMS誘變‘台農17號’及‘台農20號’鳳梨組織培養小苗之最適條件 32
試驗四、EMS誘變‘台農20號’鳳梨癒傷組織之增殖 35
試驗五、羥脯胺酸篩選‘台農17號’與‘台農20號’鳳梨組織培養小苗之最適條件 37
試驗六、低溫處理‘台農17號’與‘台農20號’ 鳳梨經EMS誘變及羥脯胺酸篩選後之組培小苗 40
五、討論 47
(一)鳳梨癒傷組織培養系統 47
(二)‘台農17號’與‘台農20號’鳳梨組織培養苗以EMS誘變之比較 49
(三)以EMS誘變‘台農20號’鳳梨癒傷組織對其增殖之影響 49
(四)羥脯胺酸與低溫逆境對鳳梨誘變品系篩選之探討 49
(五)結語 50
六、參考文獻 51

 
表目次
表1、‘台農17號’鳳梨葉片白色基部培養於不同生長調節劑培養基之誘導結果 16
表2、‘台農17號’鳳梨短縮莖培養於不同生長調節劑培養基之誘導結果 17
表3、‘台農20號’鳳梨葉片白色基部培養於不同生長調節劑培養基之誘導結果 18
表4、‘台農20號’鳳梨短縮莖培養於不同生長調節劑培養基之誘導結果 19
表5、‘台農17號’鳳梨癒傷組織於不同生長調節劑培養基培養4週後之結果 24
表6、‘台農17號’鳳梨癒傷組織於不同生長調節劑培養基培養8週後之結果 25
表7、‘台農20號’鳳梨癒傷組織於不同生長調節劑培養基培養4週後之結果 26
表8、‘台農20號’鳳梨癒傷組織於不同生長調節劑培養基培養8週後之結果 27
表9、EMS處理對‘台農17號’鳳梨組培苗存活率之影響 33
表10、EMS處理對‘台農20號’鳳梨組培苗存活率之影響 34
表11、EMS處理對‘台農20號’鳳梨癒傷組織培植體生長之影響 36
表12、不同濃度羥脯胺酸對‘台農17號’鳳梨組培苗存活率之影響 38
表13、不同濃度羥脯胺酸對‘台農20號’鳳梨組培苗存活率之影響 39
表14、低溫處理對‘台農17號’與‘台農20號’鳳梨組培苗存活率之影響 41
表15、低溫處理對‘台農17號’與‘台農20號’鳳梨組培苗脯胺酸含量之影響 42
表16、低溫處理對‘台農17號’鳳梨組培苗存活率之影響 43
表17、低溫處理對‘台農17號’鳳梨組培苗脯胺酸含量之影響 44
表18、低溫處理對‘台農20號’鳳梨組培苗之存活率 45
表19、低溫處理對‘台農20號’鳳梨組培苗脯胺酸含量之影響 46


圖目次
圖 1、‘台農17號’鳳梨組培苗葉片白色基部與短縮莖，經不同條件培養基誘導之培植體型態 20
圖 2、‘台農20號’鳳梨組培苗葉片白色基部與短縮莖，經不同條件培養基誘導之培植體型態 21
圖 3、‘台農17號’鳳梨癒傷組織培養4週後之結果 28
圖 4、‘台農17號’鳳梨癒傷組織培養8週後之結果 29
圖 5、‘台農20號’鳳梨癒傷組織培養4週後之結果 30
圖 6、‘台農20號’鳳梨癒傷組織培養8週後之結果 31

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