http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/84 2026-10-03T11:19:33Z 2026-10-03T11:19:33Z Tsung-Han Hou 侯宗翰 http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/58633 2026-07-22T11:23:59Z 2013-01-01T00:00:00Z

標題: 龍葵葉片95%酒精萃取物之抗氧化能力及抗醣化作用分析; Antioxidation and antiglycation of 95% ethanolic extracts prepared from the leaves of Solanum nigrum 作者: Tsung-Han Hou; 侯宗翰 摘要: 最終醣化蛋白(advanced glycation end-products, AGEs)的形成，導致發炎反應的發生與胰島素抗性(insulin resistance)的產生。有證據顯示抗氧化劑可以抑制活性氧(reactive oxygen species, ROS)的形成，經由抑制糖基化(glycation)減少醣化終產物的生成。龍葵(Solanum nigrum)可以作為改善血糖的藥用食物，然而其活性成分與作用機轉仍然不清楚。本研究結果顯示，龍葵95%乙醇萃取物的抗氧化活性高於龍葵50%乙醇萃取物和龍葵水萃取物。此外，龍葵95%乙醇萃取物具有抑制糖基化的活性，可以抑制果糖胺(fructosamine)與α-二羰基化合物(α-dicarbonylcompiund)的生成。在每毫克龍葵95%乙醇萃取物中，solasonine與solamargine的量分別為0.484毫克和0.183毫克。基於上述，龍葵具備當作萃取抗糖基化和抗糖尿病活性成分的潛力。; Solanum nigrum are recently not only considered to be the brand-new source of functional ingredients that exerts antiglycation and anti-diabetes activities but also to be kind of medicinal food for improving blood glucose. However, there are only few identities of the active compounds and how those above-mentioned compounds counteract diabetes are still unknown. To induces insulin resistance occurs in an inflammatory response that requires the hyperglycemia results in the formation of advanced glycation end-products (AGEs). Evidence indicates that antioxidants can suppress the formation of reactive oxygen species, decrease levels of AGEs by inhibiting glycation. In this research demonstrates that 95% ethanolic extracts of Solanum nigrum exerted significant antioxidative activity compared with 50% ethanolic extracts and aqueous extracts. Moreover, 95% ethanolic extracts of S. nigrum produced antiglycative activity, which contributed to the inhibition of fructosamine and generation of α-dicarbonyl compounds. The results indicate that 95% ethanolic extracts of S. nigrum are with great bioactivities on antiglycation and anti-diabetes.

2013-01-01T00:00:00Z Chun-Yi Hsu 徐駿逸 http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/4635 2026-07-22T11:34:31Z 2015-01-01T00:00:00Z

標題: 鼠尾草屬植物之花器生物學、生殖系統與種間雜交; The Floral Biology, Breeding System, and Interspecific Hybridization of Salvia spp. 作者: Chun-Yi Hsu; 徐駿逸 摘要: 鼠尾草屬(Salvia L.)約有900多種物種，含括一、二或多年生草本或灌木，約有140個栽培種，但僅有少數鼠尾草有商業生產或應用於景觀，可藉由種間雜交育成鼠尾草新品種。本研究以紅花鼠尾草(S. coccinea Juss. ex Murr.)、本屬少見黃花且為臺灣特有之黃花鼠尾草[S. nipponica Miq. var. formosana (Hayata) Kudo]、長蕊鼠尾草(S. patens Cav.)及一串紅(S. splendens Sell. ex Roem. & Schult)為材料，觀察花器構造、傳粉機制與花粉離體萌發。進行自交及雜交，觀察花粉管生長、小堅果及其內部發育，以瞭解本屬物種間之雜交能力。另調查種間雜交後代之性狀，供後續種間雜交選育參考。 　　紅花鼠尾草‘Snow Nymph’花朵之可育藥室距柱頭近，雄蕊無槓桿運動功能。黃花鼠尾草為典型槓桿狀雄蕊，可育藥室位於上唇瓣內，不育之藥隔下臂阻擋傳粉者取食花蜜。一串紅‘Vista Red’具可育藥室距柱頭近及遠之花朵，雄蕊無槓桿運動功能。紅花鼠尾草‘Snow Nymph’及一串紅‘Vista Red’於無傳粉者環境下可自花結實，而黃花鼠尾草則須傳粉者幫忙傳粉，此三種鼠尾草均無自交不親合性。 　　於含0.1%酪蛋白水解物之修改過Brewbaker和Kwack培養基中，添加蔗糖及聚乙二醇-3350 (polyethylene glycol-3350, PEG)可改善本屬植物花粉萌發及花粉管生長情形。紅花鼠尾草‘Snow Nymph’花粉適合添加5%-10%蔗糖+30% PEG；黃花鼠尾草花粉適合添加10%蔗糖+20% PEG；而一串紅‘Vista Red’花粉適合添加0%蔗糖+30% PEG或10%蔗糖+20% PEG，‘Vista White’花粉適合添加0%-15%蔗糖+30% PEG或10%蔗糖+20% PEG。本屬植物花粉適合之培養基滲透潛勢為-2.45 MPa至 -1.42 MPa。蔗糖有利花粉萌發但並非萌發之必要成分。 　　以含10%蔗糖+20% PEG之培養基培養花粉，紅花鼠尾草‘Snow Nymph’及一串紅‘Vista White’於開花後1天之花粉有最佳萌發率，黃花鼠尾草於開花當天有最佳萌發率，而一串紅‘Vista Red’於開花後1天至開花後2天有最佳萌發率。紅花鼠尾草‘Snow Nymph’、黃花鼠尾草及一串紅‘Vista Red’於花朵開放當天柱頭即可接受花粉，無雄蕊先熟現象。 　　觀察本屬種間雜交花粉管生長情形，以紅花鼠尾草‘Snow Nymph’、黃花鼠尾草或一串紅‘Vista Red’為種子親(♀)，與紅花鼠尾草‘Snow Nymph’ (♂)、黃花鼠尾草(♂)、長蕊鼠尾草‘Blue Angel’及一串紅‘Vista Red’ (♂)雜交，除紅花鼠尾草‘Snow Nymph’之花粉管無法穿入黃花鼠尾草之柱頭，及長蕊鼠尾草‘Blue Angel’之花粉管於授粉後2天才見於黃花鼠尾草之花柱末端外，其餘組合皆可於授粉後1天在花柱末端觀察到有花粉管螢光。種間雜交組合中，僅紅花鼠尾草‘Coral Nymph’ (♀)或‘Snow Nymph’ (♀)與一串紅‘Vista Red’ (♂)或‘Vista White’ (♂)雜交可得成熟種子；以黃花鼠尾草(♀)與長蕊鼠尾草‘Blue Angel’ (♂)或與一串紅‘Vista Red’ (♂)雜交之小堅果未膨大；多數雜交小堅果均僅部分膨大，本屬種間雜交多有受精後障礙。 　　黃花鼠尾草之雌配子體受特化珠被細胞包圍，其自交後胚及胚乳發育較雌配子體無特化珠被細胞包圍之一串紅及紅花鼠尾草慢。若種間雜交胚及胚乳敗育，則小堅果於授粉後8-16天基部褐化及/或皺縮。雜交組合中僅有以紅花鼠尾草‘Coral Nymph’或‘Snow Nymph’為種子親(♀)，與一串紅‘Vista Red’ (♂)雜交可得近成熟雜交胚，與黃花鼠尾草(♂)雜交之雜交胚可發育達球胚至心臟胚早期。以一串紅‘Vista Red’為種子親(♀)，與紅花鼠尾草‘Coral Nymph’ (♂)或‘Snow Nymph’ (♂)雜交僅得發育達魚雷期之雜交胚，與黃花鼠尾草(♂)雜交者，於授粉後4天胚及胚乳已敗育。 　　紅花鼠尾草(♀)與一串紅(♂)雜交之小苗具生長弱勢現象，本研究中僅有紅花鼠尾草‘Snow Nymph’ × 一串紅‘Vista Red’及紅花鼠尾草‘Snow Nymph’ × 一串紅‘Vista White’生長至開花階段。二種間雜交組合後代具新植株外觀及花序形態，花朵性狀多介於兩親本之間，且每輪小花數較一串紅多。雜交後代之萼片帶有灰橘、紅或紫色，花冠為橘紅色，此外，紅花鼠尾草‘Snow Nymph’ × 一串紅‘Vista White’之花瓣具白色斑塊。雜交組合後代之花粉具多形性且稔性低，經自交或與親本回交均無法結實。; Salvia L. is a large genus comprised of 900 species of annual, biennial, or perennial herbs, or shrubs. Although about 140 species are in cultivation, only a few salvias are commonly found in commerce and gardens. Interspecific hybridization could contribute more Salvia cultivars. Salvia coccinea Juss. ex Murr., yellow-flowered Taiwan-endemic S. nipponica Miq. var. formosana (Hayata) Kudo, S. patens Cav., and S. splendens Sell. ex Roem. & Schult were collected and studied. The objectives of the study were to determine the floral structure, pollination system, in vitro pollen germination, and hybridization crossability. Pollen tube and nutlet growth were observed after self- or interspecific pollination. Interspecific hybrids of S. coccinea × S. splendens were obtained, and progeny traits were investigated. 　　Observation revealed that S. coccinea ‘Snow Nymph’ had homostylic flowers and immovable stamens. Salvia nipponica var. formosana had typical lever-like stamens, with fertile theca concealed in the upper lip and the sterile lower connective arm restricted access to nectar. Salvia splendens ‘Vista Red’ had pin and homostylic flowers and immovable stamens. Salvia coccinea ‘Snow Nymph’ and S. splendens ‘Vista Red’ could set seeds without pollinators, while S. nipponica var. formosana must require pollinators for pollination. All these three salvias were self-compatible. 　　Supplement of sucrose and polyethylene glycol-3350 (PEG) in the modified Brewbaker and Kwack medium with 0.1% casein hydrolysate could improve pollen germination and tube growth of Salvia. Highest pollen germination was recorded in medium containing 5%-10% sucrose + 30% PEG for S. coccinea ‘Snow Nymph’, 10% sucrose + 20% PEG for S. nipponica var. formosana, 0% or 10% sucrose + 20% PEG for S. splendens ‘Vista Red’. However, pollen of ‘Vista White’ germinated well with media containing 0%-15% sucrose + 30% PEG or 10% sucrose + 20% PEG. The optimum osmotic potential for pollen germination ranged from -2.45 to -1.42 MPa. In these three salvias investigated, sucrose benefited but was not necessary for pollen germination. 　　Effect of flower age on pollen germination rate was measured with medium containing 10% sucrose + 20% PEG. In both S. coccinea ‘Snow Nymph’ and S. splendens ‘Vista White’, pollen had the highest germination rate at 1 day after anthesis. In S. nipponica var. formosana, pollen had the highest germination rate at anthesis. Highest germination rate of S. splendens ‘Vista Red’ was obtained at 1 and 2 d after anthesis. Stigma had became receptive at anthesis in S. coccinea ‘Snow Nymph’, S. nipponica var. formosana, and S. splendens ‘Vista Red’, indicating these plants were not protandrous. 　　Among all cross combination tested in this study, pollen tubes were observed in the style base at 1 d after pollination when S. coccinea ‘Snow Nymph’, S. nipponica var. formosana, or S. splendens ‘Vista Red’ were crossed with S. coccinea ‘Snow Nymph’, S. nipponica var. formosana, S. patens ‘Blue Angel’, or S. splendens ‘Vista Red, respectively. Pollen tubes of S. coccinea ‘Snow Nymph’ were arrested on stigma of S. nipponica var. formosana. Pollen tubes of S. patens ‘Blue Angel’ were not observed in style base of S. nipponica var. formosana until 2 d after pollination. Mature seeds were obtained only when S. coccinea ‘Coral Nymph’ or ‘Snow Nymph’ as female parents crossed with S. splendens ‘Vista Red’ or ‘Vista White’. Nutlets did not enlarge when S. nipponica var. formosana was crossed with S. patens ‘Blue Angel’ or S. splendens ‘Vista Red’. In most cross combinations, hybrid nutlets showed only partial development indicating that post-fertilization barriers existed. 　　The megagametophyte of S. nipponica var. formosana was, but those of S. splendens and S. coccinea were not, surrounded by a differentiated integumental cell layer. Selfed-embryo and endosperm of S. nipponica var. formosana developed slower than S. splendens and S. coccinea. Most hybrid embryo and endosperm aborted and resulted in basal shrinking and/or browning nutlet at 8-16 d after pollination. Nearly-mature embryo was obtained in S. coccinea ‘Coral Nymph’ or ‘Snow Nymph’ as female parent crossed with S. splendens ‘Vista Red’. Embryo reached globular to early-heart stage in S. coccinea ‘Coral Nymph’ or ‘Snow Nymph’ as female parent crossed with S. nipponica var. formosana. Embryo most reached torpedo stage in S. splendens ‘Vista Red’ crossed with S. coccinea ‘Coral Nymph’ or ‘Snow Nymph’. In contrast, embryo and endosperm aborted 4 d after pollination when S. splendens ‘Vista Red’ crossed with S. nipponica var. formosana. 　　Hybrid weakness was observed in S. coccinea × S. splendens progenies. In this study, only progenies of S. coccinea ‘Snow Nymph’ × S. splendens ‘Vista Red’ and S. coccinea ‘Snow Nymph’ × S. splendens ‘Vista White’ grew and flowered. Both hybrid progenies had new plant and inflorescence appearance. Most flower characteristic traits of both hybrid progenies were between two parents. The selected progenies had more nodal florets than S. splendens. Both hybrid progenies had green with gray-orange, red or purple calyx and orange-red corolla. Corolla had white blotch in progenies of S. coccinea ‘Snow Nymph’ × S. splendens ‘Vista White’. Both hybrid progenies had pollen polymorphism with low fertility and could not set seed after self-pollination or backcross with both parents.

2015-01-01T00:00:00Z Rong-Show Shen 沈榮壽 http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/32098 2026-07-22T09:23:18Z 2006-01-01T00:00:00Z

標題: 黛粉葉腋芽培養與體胚發生再生系統之建立; Establishment of regeneration system via axillary bud culture and somatic embryogenesis in Dieffenbachia spp. 作者: Rong-Show Shen; 沈榮壽 摘要: 為增加黛粉葉微體繁殖培植體供給來源與解決內生污染，選用兩個優良品系之成熟莖段，預處理BA進行潛伏芽體誘發，顯示潛伏芽體之誘發隨著BA濃度(0-750 mg/L)增加而遞增；誘發側芽增加數，複莖性品系(Dieffenbachia Chiada-84 ＃1012)比單莖性品系(Dieffenbachia Chiada-84 ＃0804)有較多芽體數，處理濃度以750 mg/L BA 效果最佳。而抗生素預處理對黛粉葉腋芽培植體離體培養污染之影響，未經抗生素處理者幾近全數污染。然以100 mg/L Rifampicin + 100 mg/L Gentamycin 或 100 mg/L Rifampicin + 100 mg/L Gentamycin + 150 mg/L Streptomycin組合濃度處理者，具有顯著降低污染率的效果分別為60%及50%；克服內生菌污染而建立初代無菌培養之腋芽培植體，芽體再生率僅達70%-75%，惟平均每一腋芽培植體芽體增殖數不多為1.2-1.4個，可知經抗生素處裡形態發生潛能降低，再生芽體之生長也顯現抑制現象。 而在黛粉葉無病原培植體準備、無菌培養建立與芽體增殖系統之建構顯示，莖頂分生組織培養於MS基礎培養基含1 mg/L NAA及1 mg/L TDZ培養15週後，最小之莖頂（0.1 x 0.25mm）培植體具有20％的存活率，較大之莖頂生長點（0.90 × 1.20mm）培植體，則顯現高繁殖潛力獲得12.3個再生芽體；另以4種大小黛粉葉腋芽分生組織為培植體來源，剝除3或4片芽鞘數的存活率可達30-35％，同時也顯現剝掉1片芽鞘的生長點培植體平均每一個反應培植體可達11.2個再生芽體；此等結果顯現利用腋芽生長點作為無病原培植體來源可降低黛粉葉內部感染，建立健康分生苗再生體系。此外，以人工乾旱逆境6週枝梢腋芽，進行精細解剖切取腋芽分生組織培養。顯示剝除2至4片芽鞘之生長點培植體，其存活成功率可達80-92.5％，有效降低內生病原菌感染之應用價值。另於培養方式與芽體增殖試驗可知，次級母莖培植體在TDZ濃度為1、5或10mg/L之液體培養者，顯著具有較高的芽體產量，平均芽體形成數可達5-6倍以上的繁殖潛力；而2節短莖培植於液體培養者，其芽體形成繁殖率僅達2倍左右，但芽體生長呈現較為整齊一致；提供黛粉葉微體繁殖種苗量化增殖方式之重要參考。 此外，利用短暫浸漬系統建立快速有效率的黛粉葉微枝梢大量生產體系。顯示以微體繁殖再生枝梢之2節短莖或母莖作為培植體，在MS培養基含1mg/L NAA與0.1-5.0mg/L TDZ或0.1-5.0 mg/L BA組合，在短暫浸漬系統培養容器中裝入330mL液體培養基可誘導產生大量叢生枝梢。尤其在培養基含有5mg/L TDZ之處理組，平均一個2節短莖或母莖培植體分別可得24.5及59.2個枝梢再生，此一產量為傳統固體培養微枝梢產量的3.3-8倍。利用短暫浸漬系統在微枝梢的生產上產生快速且大量的經濟效益，有助於黛粉葉微體繁殖微枝梢量產體系的建立。叢生枝梢經馴化出瓶，以500 mg/kg NAA粉劑預處理，扦插於穴盤噴霧加濕環境，枝梢長度大於3.0㎝之微插穗扦插後30天存活率達80-100％；而叢生枝梢團塊分割成1/4分割或1/2分割處理者，亦得83.3-100％存活率，並可於30天發根期長成緊密充實之株型，可利用於黛粉葉優質採穗母株建立。 另外為了了解生長調節在黛粉葉雄花序培植體體胚發生與植株再生的效果。雄花序培植體培養於修改之1/2 MS含2％蔗糖、1％葡萄糖與0.18％水晶洋菜的固體培養基，顯示在0.5 mg/L Kinetin組合2,4-D為4.0 mg/L或5.1 mg/L等觀察到較高胚狀體發生率為26.7％；根據胚狀體形成率分析黛粉葉體胚發生潛能，顯示2,4-D濃度的增加，與黛粉葉體胚發生頻率呈現顯著相關（R2=0.94）。再依前項結果修改生長素為2,4-D、 Dicamba及Picloram等生長素參試，可知Dicamba、Picloram在濃度為2mg/L培養基，培養第8週能夠獲得強胚性癒傷組織發生，其中2mg/L Dicamba達到指數 IV的胚性癒傷組織指數為72.1％為最高，在12週培養可見30％的成熟體胚形成，平均每個雄花序培植體可得22.2體胚，顯著高於其它處理別。再經修改生長調節劑以2,4-D 4mg/L組合TDZ 0.5-1.0mg/L之處理，進一步誘致高頻率球狀體胚發生率達100％，平均產生32.1-38.2個成熟體胚，少數成熟體胚偶發性轉化獲得再生小植株。 因此，為了更有效提升黛粉葉體胚發生量化增殖與胚苗轉化。雄花序培植體於半量MS基本鹽類，添加20 g/L蔗糖、10 g/L葡萄糖及1.8 g/L Gelrite等為基礎培養基，顯示在2 mg/L Dicamba及0.5 mg/L TDZ組合，具有最高初級體胚發生頻率(100％)，以及最多盾狀體胚形成數達75.2個，對黛粉葉雄花序培植體高效率體胚發生的促進，具顯著的效果。再以前一試驗均質初級體胚，作為次級培植體來源，繼代培養於相同基本培養基添加2 mg/L Dicamba及TDZ（0.5、1mg/L），培養13週後顯現最高效率(100％)的次級體胚發生頻率，次級球狀體胚形成數則達34.3及33.0個，顯示此等培養基組成，有效的促進次級體胚發生和體胚量化增殖。此外，在1/2 MS 添加150 mL/L椰子水之培養基組成，成熟次級體胚(子葉鞘胚)之胚苗轉化試驗顯示，大小約為4-5 mm之鞘葉胚，培養在0.5%葡萄糖及1.0 mg/L TDZ濃度之胚苗轉化率最高達73.3％；平均每一培植體胚苗轉化獲得之再生植株可達7.8個，且可發育成為具形態雙極性之獨立個體。胚苗轉化再生植株，經過馴化出瓶種植之穴盤苗可在溫室中表現正常生育。; In this research the influence of pathogen-free explant preparation on aseptic culture and shoot proliferation in Dieffenbachia ‘Starshine’ was study. Using different sizes of apical meristem as initial pathogen-free explants and cultured on solidified Murashige and Skoog (MS) medium with 1mg/L NAA and 1mg/L TDZ for 15 weeks. The 20% of survival rate were obtained from the minimum size (0.1x0.25mm) of shoot tip explant, and 12.3 shoots were regenerated from the larger size (0.9x1.2mm) of shoot tip explant during primary aseptic culture. Moreover, tests used 4 sizes of axillary bud meristem as initial pathogen-free explant for primary aseptic culture. Result demonstrates that removed 3 or 4 coleoptiles of axillary bud explant, gave the survival rate of 30-35%. But, on average, 11.2 shoots regenerated from excided 1 coleoptile of axillary bud explant. These results indicated using apical meristem as pathogen-free explant gave optimize information for primary aseptic culture in Dieffenbachia ‘Starshine. Furthermore, pretreatment of stock plants with 6-week drought stress and further dissected axillary bud under aseptic condition as pathogen-free explants for primary aseptic culture. Results show the explant preparation procedure gave the high survival rate was 80-92.5% from removed 2-4 coleoptiles of axillary bud explant. This result suggests using axillary bud explant dissection and treating with drought stress could escape contamination from the interior contaminants. At the same procedure, more than 90% of survival rate was observed in axillary bud explants for successfully established of primary aseptic culture between tested cultivars（D. cv. Rudolph Roehrs and D. cv. Jupiter）. In addition, the influence of culture methods and TDZ concentration on shoot multiplication were investigated. For shoot multiplication, concentration of 1, 5, and 10mg/L TDZ in the liquid culture were found that shoot yields have a significant effect achieved 5-6 folds. While, 2-nodal explant was cultured in liquid medium only obtained 2 folds of multiplication rate, but compared to solid culture, the regenerated shoots were more rapid growth and uniformity during subculture. This result suggests that liquid culture can significantly enhance axillary shoot proliferation from stock stem explants and 2-nodal explants for mass multiplication in Dieffenbachia seedlings. A rapid and efficient procedure is described for mass multiplication of microshoots using temporary immersion system (TIS) in dieffenbachia (Dieffenbachia ‘Starshine’). Multiple shoot proliferation was induced from 2-nodal and stock-stem explants on Murashige and Skoog (MS) media supplemented with 1 mg/L NAA and 0.1-5.0 mg/L thidiazuron (TDZ) or BA. Especially, the 2-nodal or stock-stem explants cultured under TIS on 5mg/L TDZ enriched medium had the highest shoot yield which was 24.5 and 59.2, respectively, the multiplication rate increased by 3.3-8 folds, over that obtained from conventional solid support systems. Therefore, the scheme for the rapid microshoot propagation of dieffenbachia using temporary immersion system had commercial efficiency. In addition, elongated multiple shoots after acclimatization, taken from the temporary immersion system, pretreated with 500mg/dm3 NAA then were cut and planted in plug with soilless mix under mist propagation condition to prevent desiccation. Survival was investigated after 30 days, microshoots taller than 3.0㎝ had the highest survival rate, which achieved 80-100%. Moreover, the shoot cluster was segmented into 1/4 or 1/2 section as propagules; the survival rate was 83.3-100%. In this production scheme, shoot cluster could be established a system of superior stock-plant for use in dieffenbachia cuttage. A method for the somatic embryogenesis and subsequent plant regeneration for Dieffenbachia ‘Tiki’ hybrids was described. Male inflorescence explants isolated from spadix of flowering plants cultured in vitro, formed embryogenic calli on surfaces of inflorescence axis within eight weeks of culture on full-strength Murashiage and Skoog (MS) medium with 4-6 mg/L 2,4-D. Somatic embryogenesis and embryoid were induced using a modified half-strength MS combination of 2 % sucrose with 1 % glucose, 0.18 % Gelrite for the basal medium, supplemented with 4.0 to 5.1 mg/L 2,4-dichlorophenoxyacetic acid (2,4-D) and 0.5 mg/L Kinetin. Male inflorescence explants transfer to modified half-strength MS basal medium with 0.5mg/L Kinetin and 4mg/L 2,4-D（or 2-4mg/L Dicamba）resulted in somatic embryogenesis at frequencies of 45-72.1% with an average of 13.3-22.2 somatic embryos per responding explant. Furthermore, at the same culture condition, male inflorescence explants were cultured onto half-strength MS basal medium with 4mg/L 2,4-D and 0.5-1.0mg/L Thidiazuron（TDZ） for embryogenesis. TDZ was found to increase the somatic embryogenesis frequencies to 100% with an average of 32.1-38.2 somatic embryos per responding explant. At the same condition, a few somatic embryos developed into complete plantlets. In this study, we are the first to report somatic embryogenesis in Dieffenbachia ‘Tiki’ and the conversion of somatic embryos to greenhouse-established plant. A regeneration system for improving the efficiency of somatic embryogenesis and emblings recovery from male inflorescences and somatic embryos of Dieffenbachia ‘Tiki’ were described. The highest percentage (100%) of primary somatic embryogenesis were achieved, and the highest yield of mature primary embryos were 75.5 on the half strength modified Murashige and Skoog（MS）medium with 2mg/L Dicamba and 0.5mg/L TDZ from male inflorescence explants. The results indicated that combination of Dicamba (2mg/L) and TDZ (0.5mg/L) significantly promoted high-efficiency multiplication of mature primary embryos in Dieffenbachia ‘Tiki’. Furthermore, the mature embryos of primary somatic embryogenesis were used as initial secondary explants for the induction of repetitive somatic embryogenesis. On the same culture medium supplemented with 2mg/L Dicamba and 0.5-1.0mg/L TDZ, the highest frequency (100%) of secondary embryo formation was obtained after 8-week subculture, and the highest number of mature secondary embryos per explant achieved 33.0-34.3 after 13-week subculture. Consequently, the same medium compositions were suitable for efficient repetitive somatic embryogenesis and multiplication of secondary embryos. In addition, emblings conversion of mature secondary embryos was investigated. The green secondary embryos of 4-5mm in size and the age of 8-10 weeks were used as explants, and cultured on half strength modified MS supplemented with 150mL/L coconut milk for somatic embryo conversion. The highest converted frequency of somatic embryo were 73.3%, and the number of converted emblings per explant reached 7.8 at 0.5% glucose and 1.0mg/L TDZ combination. Plantlets conversion from embryo was successfully acclimatized to greenhouse conditions. This technique could have significant industry application in dieffenbachia micropropagation, based on it has high efficiency of somatic embryo formation and high level of plant recovery.

2006-01-01T00:00:00Z 盧意婷 Yi-Ting Lu http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/104829 2026-09-02T16:27:18Z 2026-01-01T00:00:00Z

標題: 黑糖芭比、紅寶石與黑珍珠蓮霧採收後糖類及糖代謝與轉運基因表現之變化; Changes in sugar composition and expression of sugar metabolic and transporter genes in postharvest wax apples cv. Sugar Barbie, Ruby, and Pink 作者: 盧意婷; Yi-Ting Lu 摘要: 蓮霧(Syzygium samarangense)為臺灣重要熱帶果樹，低溫貯藏可有效延長採後壽命並抑制病害發生，但亦容易誘發寒害(chilling injury, CI)，降低果實外觀品質及商品價值。糖類除作為呼吸作用之碳源外，亦參與滲透調節、逆境訊號傳遞及碳源分配，因此糖代謝與糖轉運之調控被認為與植物低溫耐受性密切相關。本研究以黑糖芭比(Sugar Barbie, SB)、紅寶石(Ruby, RB)及黑珍珠(Pink, PK)三種不同耐寒性之蓮霧果實為材料，分別於20 ℃及5 ℃貯藏，分析貯藏期間及回溫後之生理品質、可溶性糖組成，並依據轉錄體分析結果篩選16個受低溫影響之糖代謝與糖轉運相關基因進行表現分析，以探討不同品種之低溫反應及耐寒性差異。結果顯示，5 ℃貯藏可有效抑制病害發生並降低失水率，但隨貯藏期間延長及回溫後寒害症狀明顯加劇，其中紅寶石寒害指數最高，黑糖芭比具有較佳耐寒性，回溫後寒害指數分別為SB 2.6 ± 0.97、RB 3.4 ± 0.84、PK 2.4 ± 0.97。三品種皆以果糖及葡萄糖為主要可溶性糖，其中PK鮮果之蔗糖比例最高，SB、RB及PK分別為3.18 ± 0.39%、3.42 ± 0.42％及16.47 ± 4.49%；低溫貯藏期間，各品種糖含量及還原糖含量整體變化幅度有限，而高蔗糖品種PK之蔗糖比例則隨貯藏由16.47 ± 4.49%下降至5.76 ± 0.49%。qPCR結果顯示，低溫下三品種多數糖代謝相關基因呈現相似之表現變化，蔗糖合成相關基因SsSUS2及SsSPS1於低溫期間整體呈下降趨勢，而蔗糖分解、糖解作用及糖訊號相關基因SsCWINV1、SsNINVc、SsHXK3、SsFRK7、SsPFK3及SsTPS9則受到低溫誘導增加2.38至43.62倍。糖轉運相關基因則呈現不同的反應模式，其中SsSTP13及SsUTR4受到低溫誘導，且SsSTP13於RB果肉最高增加76倍；SsSWEET15、SsERDL6及SspGlcT4整體呈下降趨勢。於所分析基因中，SsPKc及SsTST2呈現最明顯之品種差異，RB果肉於低溫貯藏第3天時SsPKc及SsTST2表現量增加至5.24及2.78倍，而SB之兩基因於貯藏期間皆未受到低溫誘導。啟動子分析顯示SB之SsPKc具有較多Dof，且SsTST2具有RB所缺乏之bZIP；RB則具有SB所缺乏之WRKY、ERF及NAC等逆境相關轉錄因子結合位點。綜合生理、糖類組成、基因表現及啟動子分析結果，不同品種蓮霧之耐寒性差異與糖類大量累積、糖代謝、糖轉運及其低溫轉錄調控方式有關，其中SsPKc及SsTST2可能為品種間低溫反應差異之關鍵基因。未來可進一步結合代謝分析及功能驗證，以釐清糖類區室間分配及糖解作用末端調控之分子機制。; Wax apple (Syzygium samarangense) is an important tropical fruit crop in Taiwan. Although low-temperature storage effectively prolongs postharvest shelf life and suppresses disease development, it also induces chilling injury (CI), resulting in deterioration of fruit appearance and market quality. In addition to serving as respiratory substrates, soluble sugars participate in osmotic regulation, stress signaling, and carbon partitioning, and the regulation of sugar metabolism and sugar transport is therefore considered closely associated with chilling tolerance. In this study, three wax apple cultivars with different chilling tolerances, Sugar Barbie (SB), Ruby (RB), and Pink (PK), were stored at 20 °C and 5 °C. Physiological quality and soluble sugar composition during storage and after rewarming were analyzed, and 16 cold-responsive genes related to sugar metabolism and sugar transport were selected based on transcriptome analysis and their expression patterns were analyzed to investigate cultivar-specific cold responses and differences in chilling tolerance. The results showed that storage at 5 ℃ effectively reduced disease incidence and weight loss, but chilling injury became more severe with prolonged storage and after rewarming. RB exhibited the highest chilling sensitivity, whereas SB showed superior chilling tolerance, with chilling injury indices after rewarming of 2.6 ± 0.97, 3.4 ± 0.84, and 2.4 ± 0.97 for SB, RB, and PK, respectively. Fructose and glucose were the predominant soluble sugars in all three cultivars, and the sucrose proportions in fresh fruit were 3.18 ± 0.39%, 3.42 ± 0.42％, and 16.47 ± 4.49% in SB, RB, and PK, respectively, with PK showing the highest proportion. During cold storage, soluble sugar and reducing sugar contents showed only limited overall changes, whereas the sucrose proportion in the high-sucrose cultivar PK declined from 16.47 ± 4.49% to 5.76 ± 0.49%. qPCR analysis showed that most sugar metabolism-related genes exhibited similar responses among the three cultivars. The sucrose biosynthesis-related genes SsSUS2 and SsSPS1 were generally downregulated during cold storage, whereas SsCWINV1, SsNINVc, SsHXK3, SsFRK7, SsPFK3, and SsTPS9 showed 2.38- to 43.62-fold induction at 5 ℃. In contrast to the generally similar responses of sugar metabolism-related genes, sugar transporter-related genes exhibited more variable expression patterns. SsSTP13 and SsUTR4 were generally upregulated during cold storage, and SsSTP13 expression increased by up to 76-fold in RB pulp, while SsSWEET15, SsERDL6, and SspGlcT4 were generally downregulated. Among the genes analyzed, SsPKc and SsTST2 showed the most pronounced cultivar differences. In RB pulp, the expression of SsPKc and SsTST2 reached 5.24- and 2.78-fold, respectively, on day 3 at 5 ℃, whereas neither gene was induced in SB during storage. Promoter analysis further revealed cultivar-specific differences in predicted transcription factor (TF) binding sites. The SsPKc promoter of SB contained more Dof sites, and the SsTST2 promoter of SB contained bZIP sites that were absent from RB; by contrast, RB contained stress-related WRKY, ERF, and NAC binding sites that were absent from SB. Collectively, the physiological, sugar composition, gene expression, and promoter analyses suggest that differences in chilling tolerance among wax apple cultivars are associated with substantial sugar accumulation, sugar metabolism, sugar transport, and cold-responsive transcriptional regulation. SsPKc and SsTST2 may represent key genes underlying cultivar-specific chilling responses. Future studies integrating metabolic analyses and functional validation may help clarify the molecular mechanisms governing subcellular sugar partitioning and regulation of the terminal step of glycolysis.

2026-01-01T00:00:00Z