臺灣博碩士論文加值系統

彩色甜椒為高價之新興蔬菜作物，唯普遍不具耐熱性，故不利於台灣周年經濟栽培。本研究之目的，擬藉由彩色甜椒耐熱品系之選拔與耐熱性之評估，以提供耐熱性彩色甜椒育種之利用。初步，本研究以自國內外收集的57個彩色甜椒品系為供試材料，經93年夏作及秋作初步篩選24個生長勢強、高結果率、果色與果型特殊者及種子採種量高之品系。再經94年夏作及秋作進行第二次篩選，依其園藝性狀篩選出果實具有經濟利用價值，並可能具有耐熱潛力之8個彩色甜椒品系，包括4個“短筒鐘型”品系(bell pepper)與4個“非短筒鐘型”品系 (non-bell pepper)。4個“短筒鐘型”品系為‘KC104’、‘C00947’、‘C01336’與‘RB102’，平均夏作之果實重量為54公克至68公克，平均單株結果數為4至7個果實；其中夏作果重與單株產量表現最高的品系為‘KC104’，夏、秋果減重率最少的品系為‘C00947’(19.7%)，為較穩定之品系。篩選出的4個 “非短筒鐘型”品系為‘C02410’、‘C00611A’、‘C02080’與‘C03338B’，均為“匈牙利”甜椒(paprika)，其夏季果實重量為30公克至38公克，單株結果數為9至14個果實；其中‘C02410’之夏、秋果實減重率為17.39%，是較穩定之品系。
以分子標誌系統化的進行作物遺傳歧異性分析，有助於作物育種效率之提升。本試驗中利用 random amplified polymorphic DNA (RAPD)、inter simple sequence repeat (ISSR) 與simple sequence repeat (SSR) 三種分子標誌進行36個番椒 (Capsicum annuum ) 品系之遺傳歧異性分析。試驗結果顯示，自26個RAPD引子反應獲得302個核酸片段產物，其中233個核酸片段具多型性 (polymorphism 77.2%)；自10個ISSR 引子反應獲得160個核酸片段，其中142個核酸片段具多型性(polymorphism 88.8%)；自12對具多型性之SSR引子組的反應產物為 61個alleles，平均 polymorphic index content (PIC) 為 0.569。合併 RAPD與ISSR 分子標誌進行UPGMA群集分析，結果顯示不同群集可對應出不同番椒果形差異；而合併 RAPD、ISSR 與SSR等三種分子標誌之UPGMA分析結果顯示，不同群集仍對應不同果形外，針對夏季產量較高之品系亦可區別。以上三種分子標誌再以二維主成份分析 (2D principle component analysis, 2D-PCA) 分析結果與 UPGMA 分析符合，且夏季產量較高之品系，亦可獲得標定。綜合試驗結果顯示，合併多種分子標誌之分析可提供較高之多型性，有利於相近品種間之遺傳分析，可作為品種選育之參考。
為評估彩色甜椒在高溫逆境下之光合作用表現，以12個彩色甜椒品系包含：田間篩選推測可能具耐熱潛力的6個“短筒鐘型”彩色甜椒品系、3個“匈牙利”甜椒品系；加上夏作結果較差的2個“短筒鐘型”彩色甜椒品系與1個商業品種，藉葉綠素螢光分析進行光合效率與耐熱性之評估。試驗結果顯示，於溫室適溫(27/22℃)下，利用葉綠素螢光分析的最小螢光值(Fo)參數與PSⅡ光化學反應最大螢光值(Fv/Fm)參數，可評估出光合作用活性表現較高之品系為‘C02080’與‘RB102’。而在花芽分化前期至始花期(42~62DAS)經高溫逆境(33/22℃)，利用葉綠素螢光分析的最小螢光值(Fo)參數、PSⅡ光化學反應最大螢光值(Fv/Fm)參數以及最大螢光值(Fm)三參數，綜合評估出高溫下合作用活性表現較高之品系為‘C02080’與‘C00611A’；而 33/27℃高溫逆境下，利用葉綠素螢光分析的最大螢光值(Fm)參數，評估出高溫下合作用活性表現較高之品系為‘C01184’。綜合適溫(27/22℃)與高溫逆境(33/22℃)下的葉綠素螢光評估結果，以‘C02080’光合效率表現最好，且可以呼應其高溫下之結果表現。
高溫環境下，常導致彩色甜椒的落花與落果現象之發生。為評估高溫對彩色甜椒授粉之影響，試驗中以12個彩色甜椒品系(包含田間篩選可能具耐熱潛力的6個 “短筒鐘型”品系與3個 “匈牙利”紅椒品系)為材料，探討高溫(33℃) 對花粉發芽活力之影響，以及生殖生長期高溫 (33/27℃)，對花粉授粉能力與雌蕊受精能力之影響，以瞭解高溫對彩色甜椒之生殖生理及結實的關係。試驗結果顯示，高溫(33℃)下彩色甜椒的花粉發芽率明顯下降，除‘Chocolate Miniature Bell’、‘C02080’、‘C05464A’與‘麗妃星’，其餘花粉發芽率皆低於5％。以螢光顯微鏡觀察高溫對花粉管in vivo生長之影響發現，高溫(33℃/27℃)下分化的花粉授粉力明顯降低，但高溫分化的雌蕊柱頭及花柱之受精能力較不受影響。進一步比較不同品系的彩色甜椒在高溫下之結果情形與果實內種子發育之數量發現，其結果與高溫下之花粉發芽率之情形，以及高溫授粉後之花粉管生長的螢光顯微鏡觀察結果一致。綜合評估高溫下花粉發芽活力與授粉結果，以‘Chocolate Miniature Bell’、‘C02080’與‘C00947’等3品系，應屬耐熱性較佳之品系，其中又以‘C02080’為耐熱性表現最好。
綜合上述研究結果顯示，以“匈牙利”彩色甜椒品系之耐熱性表現，較優於“短筒鐘型”彩色甜椒，試驗中針對高溫下的光合作用效率、授粉與結果表現，均以“匈牙利” ‘C02080’之耐熱性表現最好；但“短筒鐘型”彩色甜椒果形則較受市場喜好，故栽培品種(系)較多，且具較高之多樣性，研究中以RAPD (random amplified polymorphism DNA)、ISSR (inter-simple sequence repeat)與SSR (simple sequence repeat)分子標誌分析結果發現， “短筒鐘型”品系間的遺傳歧異性較高，果重差異較大(14~153公克)，比較高溫下的花粉活力或光合作用效率表現，則以中型果‘C00947’(64公克)表現較佳，僅次於‘C02080’品系。研究結果顯示‘C02080’與‘C00947’為較具潛力之彩色甜椒品系。In vitro高溫下之花粉發芽表現，或適溫下花芽分化至始花期間之光合效率，可用以輔助彩色甜椒耐熱性之評估。

Since colored sweet pepper has high economic value, it without the habit of heat-tolerance can not be year-round cultured in Taiwan. The purposes of this study were to select and evaluate the heat-tolerance of color sweet pepper, providing for the heat-tolerance breeding program. First, a total of 57 colored sweet pepper genotypes collected from homeland and abroad had been used as materials in this study. Among them, 24 genotypes were selected based on vigor growing, high fruiting rate, better fruit color and fruit type, and high seed amount by field evaluating in summer and autumn trials 2004. Eight genotypes had then been selected on account of better economic values, good horticultural properties and high heat-tolerance potentiality by field evaluating in summer and autumn trials 2005. They included four genotypes of bell pepper and four genotypes of non-bell pepper. Data obtained from summer trials showed that the averaged fruit weight of selected four bell pepper genotypes (‘KC104’, ‘C00947’, ‘C01336’ and ‘RB102’) was from 54 to 68 g, and the averaged fruit number per plant was from 4 to 7 fruits. Among them, ‘KC104’ had the highest fruit yield in summer trial, particularly. ‘C00947’ with the medium-sized fruit and having a least fruit weight difference of 19.7% performed as more stable genotype than others in the summer and autumn trials. Data obtained from summer trials also showed that the fruit number per plant of selected four non-bell pepper, i.e. paprika pepper, (‘C02410’, ‘C00611A’, ‘C02080’ and ‘C03338B’) was from 9 to 14 fruits per plant and the fruit weight was from 30 to 38 g. The fruit weight difference (%) between summer and autumn trials was from 17.39% to 62.00%. ‘C02410’ with a weight difference of 17.39% performed more stable in the summer and autumn trials.
Assessment of the genetic diversity by molecular markers is useful for their systematic and high efficiency for crop breeding program. There were 36 genotypes of Capsicum, including 34 genotypes of sweet pepper and 2 genotypes of chili pepper which were conducted by random amplified polymorphic DNA (RAPD), inter simple sequence repeat (ISSR) and simple sequence repeat (SSR) to estimate pair wise genetic similarity. The result showed 302 RAPD fragments were generated from 12 primers, and 233 with polymorphism (77.2%). For ISSR analysis, 160 fragments were obtained from 10 primers, and 142 with polymorphism (88.8%). For SSR assay, 61 alleles were generated and the polymorphic index content (PIC) 0.569. Furthermore, a combination of RAPD and ISSR markers assessed the cluster analysis of UPGMA, showed the relationships among groups following the fruit shapes. While combined all of the RAPD, ISSR and SSR markers also reveled the groups generally cluster together with similar fruit sharps even the high yield genotypes in summer would be distinguished too. Combining these three marker systems and assessed principle component analysis (PCA) that resulted as well as the UPGMA analysis, besides the higher yield genotypes in summer could be distinguished clearly. These results suggest that a combination of different kinds of markers can generate sufficient polymorphism to assess genetic variation even within closely related species.
For evaluating the photosynthesis efficiency of color sweet pepper under heat stress, using 12 genotypes of colored sweet pepper selected in the field, including 6 genotypes of bell-pepper and 3 of paprika pepper that might have heat-tolerance potentiality; two additional genotypes of bell-pepper performed a little worse in summer test and one commercial variety, were examined to evaluate the efficiency of photosynthesis and heat–tolerance of colored sweet pepper utilizing the chlorophyll fluorescence assessment. Under environment temperature of 27/22℃, the experimental results showed ‘C02080 ’and ‘RB102’ had higher performance of photosynthesis efficiency among 12 genotypes of colored sweet pepper which evaluated by chlorophyll fluorescence parameters of Fo (minimal of fluorescence) and Fv/Fm (Maximal quantum efficiency of PSⅡ photochemistry). For treatment of high-temperature circumstance (D/N: 33/22℃) from flower differentiation stage to anthesis (42~62 DAS), fluorescence parameters of Fm (maximal of fluorescence), Fo and Fv/Fm could be used as good parameters to evaluate the photosynthesis efficiency among the tested genotypes, and the results showed ‘C02080’ and ‘C00611A’ with the better performance. While under treatment of 33/22℃ and evaluated by Fm parameter, the best performance was ‘C01184’. The parameters of chlorophyll fluorescence assessment under temperature of 27/22℃ and 33/22℃ may respond to the behavior of the colored sweet pepper when growing under high-temperature circumstance. Screened genotype of ‘C02080’ with vigor growing and having heat-tolerance potentiality in the field also appeared with the best photosynthesis efficiency by the chlorophyll fluorescence analysis.
High temperature would cause flowers and fruits drop in sweet pepper. In order to assess the heat tolerance behavior of different genotypes of the colored sweet peppers, 12 genotypes, including 6 bell-pepper genotypes and 3 of paprika pepper that have been screened in the field and might have heat-tolerance potentiality, had been investigated under high temperature (33 degrees Centigrade) with an aim to evaluate the influence of pollen vigor in vitro and under another high temperatures (33/27oC) within the reproduction growth period to evaluate the performances of pollination ability of pollen and fertilization ability of pistil. Furthermore, the result of the experiments indicated that high temperature (33oC) obviously reduced the pollen germination rates of tested colored sweet peppers. Except for ‘Chocolate Miniature Bell’, ‘C02080’, ‘C05464A’, and ‘Beauty Star’, the pollen germination percentages of other tested genotypes were all lower than 5%. In addition, the influence of the high temperatures (33/27 ℃) on the growth of pollen tubes was observed by the fluorescent microscope. High temperatures obviously reduced the pollination ability and pollen tube growth, but the fertilizing ability of pistil was not influenced. The tendency of the fruiting habits and the fruit quantity of tested genotypes under high temperatures were the same as the performances of pollen germination rates and the growing behavior of pollen tubes under the same condition. The assessment results of pollen vigor and pollinating under high temperatures revealed ‘Chocolate Miniature Bell’, ‘C02080’, and‘C00947’ should belong to the genotype with better heat-tolerance; however, ‘C02080 ’was the best among them.
After generalized assessment of study results described above, the heat tolerance performance of paprika type of colored sweet pepper was better than the bell colored sweet pepper. Experimental results showed that ‘C02080’ (paprika type) performed best in the photosynthesis efficiency, pollinating behaving, and pollination results under the high temperature circumstances; however, the bell colored sweet pepper with good at fruit shape and having large diversified cultivars was relatively popular by the market. Through assessment data of RAPD (random amplified polymorphism DNA), ISSR (inter-simple sequence repeat), and SSR (simple sequence repeat) approved that genetic diversity of the bell colored sweet pepper was indeed high. For example, the fruit weigh differed relatively large between 14g to153 g. Finally, compared with the pollen vigor or photosynthesis efficiency under high temperature circumstances, ‘C00947’ with the medium-sized fruit (64 g) performed better than others genotype, but second to ‘C02080’. All of results revealed that ‘C02080’ and ‘C00947’ both were the most superior colored sweet pepper genotype with heat tolerance abilities. These also revealed that the performances of pollen germination under high temperature in vitro, and photosynthesis efficiency from earlier flowers differentiation stage to beginning of anthesis could be used as useful tools for assessment of heat tolerance evaluation for colored sweet pepper.

中文摘要••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••i
英文摘要•••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••v
緒言••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••viii
第一章、前人研究••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••1-1
一、甜椒(Capsicum annuum. L)之馴化栽培與沿革••••••••••••••••••••••••••••••••••••1-1
二、核酸分子標誌與作物之遺傳研究•••••••••••••••••••••••••••••••••••••••••••••••••••••••••1-3
三、高溫逆境與植物光合反應之研究•••••••••••••••••••••••••••••••••••••••••••••••••••••••••1-5
四、溫度對甜椒開花、授粉與結果生理之影響•••••••••••••••••••••••••••••••••••••••••••1-8
第二章、彩色甜椒之遺傳表現型與耐熱性差異之評估••••••••••••••••••••••••••••••••••••••••••••••2-1
中文摘要••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••2-1
英文摘要••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••2-2
前言••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••2-3
材料與方法••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••2-4
結果••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••2-5
討論••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••2-29
附錄••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••2-31
第三章、分子標誌應用於彩色甜椒遺傳差異性之研究••••••••••••••••••••••••••••••••••••••••••••••3-1
中文摘要••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••3-1
英文摘要••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••3-2
前言••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••3-3
材料與方法••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••3-4
結果••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••3-6
討論••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••3-31
第四章、利用葉綠素螢光反應評估彩色甜椒之耐熱性••••••••••••••••••••••••••••••••••••••••••••••4-1
中文摘要••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••4-1
英文摘要••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••4-2
前言••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••4-3
材料與方法••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••4-4
結果••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••4-5
討論••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••4-29
第五章、高溫對彩色甜椒花粉活力與授粉之影響••••••••••••••••••••••••••••••••••••••••••••••••••••••5-1
中文摘要••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••5-1
英文摘要••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••5-2
前言••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••5-3
材料與方法••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••5-4
結果••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••5-7
討論••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••5-22
參考文獻••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••6-1

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