臺灣博碩士論文加值系統

茄子為世界重要蔬菜作物之一，且有益於人體健康，有許多關於透過組織培養誘導茄子植株再生的研究，研究指出運用不同植物賀爾蒙以及不同的茄子培植體(葉片、根段、子葉、胚軸或節間)可誘導植株再生，雖然如此，在不同光照強度下，以植物生長調節劑誘導茄子雜交種之植株再生的研究有限，因此在本實驗以不同的種類物生長調節劑，生長素Auxin (naphthaleneacetic acid, NAA)、 細胞分裂素Cytokinin (6-benzyladenine, BA)以及 Thidiazuron (TDZ)在不同光強度下，以誘導茄子雜交種葉片培植體之植株再生，以NAA或BA誘導其葉片再生之效果不顯著， 而TDZ對芽體再生之效果較NAA及BA佳，若以0.1 mg/L TDZ在正常光照強度(35μmol/m2/s)處理葉片培植體21天，平均每培植體有4.94 ± 1.84個芽體，另外一方面，以0.1 mg/L及0.25mg/L TDZ 處理葉片培植體(5 mm2)有較多的芽體數，每培植體平均分別有11.19 ± 2.54及12.30 ± 3.30個芽體，而葉片培植體(5 mm2)在低光條件(13 mol/m2/s)下處理0.1 mg/L TDZ有最多的芽體數，平均每個培植體有15.41±0.71個芽體數。本實驗以組織切片方法觀察芽體發育的形態，結果顯示其芽體為器官再生之不定芽，植株再生方面，先前在低光條件下以0.1 mg/L TDZ處理之葉片培植體繼代
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到不含植物生長調節劑之MS培養基中，培養42天有明顯生長及發育的情形，而在高光及黑暗之環境條件下則否，另外雜交種上位葉培植體在低光條件下及TDZ 處理後，繼代於MS培養基中不定芽有最高的存活率，本實驗找出較適當光照條件以及TDZ濃度以誘導茄子雜交種葉片培植體之不定芽的發生，未來可以TDZ誘導其他茄子雜交種之植株再生。

Eggplant (Solanum melongena L.) is a vegetable and medicinal plant good for human health; furthermore, eggplant is an important crop all over the world. A number of studies report the regeneration of large number eggplants through tissue culture; for example, researchers would add hormone in the tissue culture media to induce the plant regeneration. Many reports were also indicated that using different explants of eggplant such as the leaves, roots, cotyledons, hypocotyls, and nodes, to induce plant regeneration. However, the studies which induce regeneration of eggplant interspecific hybridization varieties with various light intensity condition were limited. Consequently, in our experiment will use leaf explants of eggplant treating with artificial hormones, such as Auxin (naphthaleneacetic acid, NAA), Cytokinin (6-benzyladenine, BA) and Thidiazuron (TDZ) and with different light intensity conditions to induce plant regeneration. In the study, the effect of leaf explants treated with NAA and BA to induce shoot regeneration eggplant interspecific hybridization varieties were not significantly. On the other hands,
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TDZ have higher efficiency shoot regeneration rates than NAA and BA. The in vitro shoot were obtained in MS medium supplemented with 0.1 mg/L TDZ, having an average shoot number of 4.94 ± 1.84 per leaf explants (not fix size) in normal light intensity (35μmol/m2/s) for 21 days; on the other hand, the leaf explants (5 mm2) cultured in MS medium supplemented with 0.1 mg/L or 0.25 mg/L TDZ which in vitro shoot number was more leaf explants than that none fixed size, having average 11.19 ± 2.54 and 12.30 ± 3.30 per explants, respectively. The best in vitro shoot induction was achieved on MS medium supplemented with 0.1 mg/L TDZ, with an average of 15.41±0.71 per leaf explants (0.5mm2) in low light intensity (13 mol/m2/s). In our experiment, histology analyzing indicated that the in vitro shoot was through the organogenesis adventitious shoot. Finally, the adventitious bud inducing from the leaf explants treated with 0.1 mg/L TDZ in low light intensity would be growing and development after subculture to MS medium with hormone free in the normal light intensity condition for 42days. Furthermore, the upper leaf explants have highest survival rates after subculture to the MS medium for 42 days. On the other hand, the adventitious shoot inducing from the leaf explants all of position wouldn’t be growing and development after subculture to the MS medium without any plant growth regulator for 42 days; moreover, that have lowest survival rate. We were fined the optimal light intensity and concentration of artificial hormones, TDZ, to induce leaf explants of eggplant regeneration. The future we can try to use TDZ to induce regeneration of other eggplant interspecific hybridization varieties in the future.

目錄
摘 要 .............................................................................................................. I
Abstract ......................................................................................................... III
誌謝 ............................................................................................................... V
目錄 ............................................................................................................. VII
圖表目錄 ...................................................................................................... IX
1 前言 ......................................................................................................... 1
2 前人研究 ................................................................................................. 2
2.1 茄子簡介 ................................................................................................. 2
2.2 組織培養之利用 ..................................................................................... 2
2.3 型態發生 ................................................................................................. 3
2.4 誘導因子 ................................................................................................. 4
2.4.1 基因型 ....................................................................................... 5
2.4.2 培植體 ....................................................................................... 5
2.4.3 植物生長調節劑 ........................................................................ 6
2.4.3.1 TDZ誘導植株再生 ....................................................... 6
2.4.4 其他影響因子 ............................................................................ 8
2.4.4.1 培植體部位及方向 ....................................................... 8
2.4.4.2 光照 ............................................................................... 8
2.4.4.3 營養元素及碳源 ........................................................... 9
2.4.4.4 多胺與體胚 ................................................................... 9
2.5 茄子的型態發生 ................................................................................... 10
3 材料方法 ............................................................................................... 13
3.1 植物材料 ............................................................................................... 13
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3.2 NAA、BA處理對茄子雜交種‘W10 × T (D)’型態發生之影響 ....... 13
3.3 TDZ誘導茄子雜交種‘W10 × T (D)’不定芽再生 .............................. 13
3.4 TDZ處理14天之葉片培植體繼代於BA培養基 ............................ 13
3.5 葉片培植體對芽體發生之影響 ........................................................... 14
3.6 葉片培植體以不同光度及TDZ處理對芽體發生之影響 ................. 14
3.7 石蠟切片 ............................................................................................... 14
3.8 不定芽及不定根之生長 ....................................................................... 15
3.9 數據分析 ............................................................................................... 15
4 結果討論 ............................................................................................... 16
4.1 NAA、BA處理對茄子雜交種‘W10 × T (D)’型態發生之影響 ....... 16
4.2 TDZ誘導茄子雜交種‘W10 × T (D)’不定芽再生 .............................. 17
4.3 TDZ處理14天之葉片培植體繼代於BA培養基 ............................ 18
4.4 葉片培植體對芽體發生之影響 ........................................................... 19
4.5 葉片培植體以不同光度及TDZ處理對芽體發生之影響 ................. 21
4.6 組織切片 ............................................................................................... 23
4.7 不定芽及不定根之生長 ....................................................................... 24
5 結論 ....................................................................................................... 26
6 參考文獻 ............................................................................................... 65
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圖表目錄
圖 1. 不含植物生長調節劑之MS培養基處理茄子種間雜種‘W10 × T (D)’根段、節間及葉片培植體第0天。 ................................................. 27
圖 2. 不同濃度 NAA處理茄子種間雜種‘W10 × T (D)’根段、節間及葉片培植體35天，較低濃度 NAA 處理下有不定根發生。 ................ 28
圖 3. 不同濃度 BA處理茄子種間雜種‘W10 × T (D)’根段及節間培植體35天，並沒有芽體之器官發生。 ..................................................... 30
圖 4. 不同濃度BA處理茄子種間雜種‘W10 × T (D)’葉片培植體35天，切口稍有緊實的癒傷組織。 .............................................................. 31
圖 5. 以不同植物生長調節劑處理茄子種間雜種‘W10 × T (D)’根、節間及葉片培植體40天，NAA處理其培植體有褐化情形，BA處理有些許不定芽發生。 ..................................................................................... 32
圖 6. 0.5 mg/L TDZ處理茄子種間雜種‘W10 × T (D)’的節間及葉片培植體。 ............................................................................................................ 33
圖 7. 不同濃度TDZ處理茄子種間雜種‘W10 × T (D)’葉片培植體。 .... 34
圖 8. 以0.1 mg/L TDZ處理茄子種間雜種‘W10 × T (D)’葉片培植體，有芽體發生。 ......................................................................................... 35
圖 9. 不同TDZ濃度處理茄子種間雜種‘W10 × T (D)’葉片培植體第21及42天，對照組葉片有不定根的發生，不同TDZ濃度處理下葉片有芽體再生。 ......................................................................................... 36
圖 10. 以0.1 mg/L TDZ處理茄子種間雜種‘W10 × T (D)’葉片培植體14天候，繼代至不同濃度BA培養基處理30天，隨著BA之處理濃度葉片培植體有不同程度之型態發生。 .............................................. 37
圖 11. 不同濃度TDZ處理茄子種間雜種‘W10 × T (D)’葉片培植體。 .. 39
圖 12. 不同濃度TDZ處理茄子種間雜種‘W10 × T (D)’葉片培植體第28天，隨TDZ濃度不同其芽體發生有不同之情況。 ......................... 40
圖 13. 在不同光照度下以0.1 mg/L TDZ處理茄子種間雜種‘W10 × T (D)’上位葉片培植體(size = 0.5 mm2)。 ................................................... 41
圖 14. 在不同光照度下以固定濃度0.1 mg/L TDZ處理茄子種間雜種‘W10 × T (D)’中位葉片培植體(size = 0.5 mm2)。 ............................ 42
圖 15. 在不同光照度下以固定濃度0.1 mg/L TDZ處理茄子種間雜種
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‘W10 × T (D)’下位葉片培植體(size = 0.5 mm2)。 ............................ 43
圖 16. 在不同光照度下以固定濃度0.1 mg/L TDZ處理茄子種間雜種‘W10 × T (D)’上位葉片培植體(size = 0.25 mm2)。 .......................... 44
圖 17. 在不同光照度下以固定濃度0.1 mg/L TDZ處理茄子種間雜種‘W10 × T (D)’中位葉片培植體(size = 0.25 mm2)。 .......................... 45
圖 18. 在不同光照度下以固定濃度0.1 mg/L TDZ處理茄子種間雜種‘W10 × T (D)’下位葉片培植體(size = 0.25 mm2)。 .......................... 46
圖 19. 不同光照度下以0.1 mg/L TDZ處理茄子種間雜種‘W10 × T (D)’大葉片培植體，隨處理時間芽體生長之情形。 .................................. 47
圖 20. 在不同光照度下以0.1 mg/L TDZ處理茄子種間雜種‘W10 × T (D)’葉片培植體，其芽體再生情形。 ...................................................... 49
圖 21.以0.1 mg/L TDZ 處理茄子種間雜種‘W10 × T (D)’葉片培植體(大)14天。 ......................................................................................... 51
圖 22. 以TDZ處理茄子種間雜種‘W10 × T (D)’葉片培植體(小) 21天。 ............................................................................................................ 52
圖 23. 在正常光照條件下，以0.1 mg/L TDZ處理茄子種間雜種‘W10 × T (D)’大葉片培植體之組織切片圖。 ................................................... 54
圖 24. 在不同光照環境下以0.1mg/L TDZ處理茄子種間雜種‘W10 × T (D)’大葉片培植體35天候繼代至不含植物生長調節劑MS培養基及一般光照環境中培養42天，不定芽生長之情形。 ......................... 55
圖 25. 在不同光照環境下以0.1mg/L TDZ處理茄子種間雜種‘W10 × T (D)’大葉片培植體35天候繼代至不含植物生長調節劑MS培養基及一般光照環境中培養60天，不定根生長之情形。 ......................... 56
表 1. 以不同濃度TDZ處理茄子種間雜種‘W10 × T (D)’葉片培植體之變方分析。 ............................................................................................. 57
表 2. 以不同濃度TDZ處理茄子種間雜種‘W10 × T (D)’葉片培植體第21天及42天芽體發生率及芽數之結果。 ............................................ 58
表 3. 以0.1 mg/L TDZ處理茄子種間雜種‘W10 × T (D)’葉片培植體14天後繼代至不同濃度之BA培養基中觀察其芽數變化。 ................... 59
表 4. 以不同濃度TDZ處理茄子種間雜種‘W10 × T (D)’葉片培植體第14、21天及28天培植體之芽體形成率。 ....................................... 60
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表 5. 以不同濃度TDZ處理茄子種間雜種‘W10 × T (D)’葉片培植體第14、21天及28天芽體發生數之結果。 ................................................... 61
表 6. 以0.1 mg/L TDZ處理茄子種間雜種‘W10 × T (D)’葉片培植體第35天，不同因子對芽數影響之變方分析。 .......................................... 62
表 7. 以0.1 mg/L TDZ處理茄子種間雜種‘W10 × T (D)’葉片培植體，不同影響因子下芽數及癒傷組織之影響情形。 .................................. 63
表 8. 茄子種間雜種‘W10 × T (D)’大葉片培植體在不同光照環境下處理後繼代至一般光照之MS培養基中培養42天。 ............................. 64

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