Dissertations / Theses on the topic 'Papaya seeds'

Trabalho Complementar apresentado à Universidade Fernando Pessoa como parte dos requisitos para obtenção do grau de licenciada em Ciências da Nutrição
O interesse pelo reaproveitamento dos resíduos alimentares obtidos das frutas e vegetais tem vindo a aumentar nas últimas décadas, não só na defesa do conceito de sustentabilidade como também pelas reconhecidas propriedades biológicas associadas ao aporte nutricional e composição de fitoquímicos descritas nestes sub-produtos. As sementes da papaia são um exemplo. De facto, da fruta, apenas se consome a polpa, sendo a casca e as sementes consideradas sub-produtos que poderão trazer um valor acrescido às indústrias alimentar, farmacêutica e cosmética. Por estes motivos, este trabalho experimental teve como principal objetivo efetuar uma quantificação do teor de fenólicos totais e de flavonoides totais, das sementes da Carica papaya Linn, bem como, avaliar a atividade antioxidante das mesmas. Os resultados obtidos revelaram que o extrato aquoso foi o que permitiu obter maior teor de flavonoides (63,24 mg EC/100g). Em contrapartida, o extrato etanólico foi o que obteve maior teor de fenólicos (981,84 mg EAG/100g). A atividade antioxidante foi avaliada através de dois métodos espectrofotométricos: DPPH• e FRAP. Por ser um antioxidante natural, o uso do ácido ascórbico como controlo permitiu medir a atividade antioxidante dos compostos bioativos presentes nos dois extratos, no sentido de os comparar com um antioxidante natural. Apesar da atividade antioxidante do ácido ascórbico ter sido superior aos dois extratos, pelos dois métodos espectrofotométricos, estes poderão ser utilizados como antioxidantes naturais, sendo que é através do extrato etanólico que podemos tirar maior partido da atividade antioxidante dos compostos bioativos das sementes da Carica papaya L. (416,87 μg/ml pelo DPPH• e 283,52 μg/ml pelo FRAP). Com os resultados obtidos podemos ainda concluir que o teor de compostos bioativos é influenciado por diversos fatores, nomeadamente, o tipo de solvente extrator utilizado. Apesar das sementes da papaia poderem ser utilizados como antioxidantes naturais, tanto na indústria alimentar, farmacêutica e cosmética, torna-se necessário aprofundar as investigações no sentido de avaliar a toxicidade das mesmas.
The interest in the reuse of food residues obtained from fruits and vegetables has been increasing in recent decades, not only in defense of sustainability but also for the recognized biological properties associated with the nutritional contribution and composition of phytochemicals described in these by-products. Papaya seeds are one of these examples. In fact, only the pulp is consumed in the fruit, the bark and seeds are considered as by-products that can bring added value to the food, pharmaceutical and cosmetic industries. For these reasons, the main objective of this experimental work is to quantify the total phenolic and total flavonoid content in Carica papaya seeds, as well as to evaluate the antioxidant activity of the seeds. The results obtained show that the aqueous extract allows to obtain a higher flavonoid content (63.24 mg CE/100g). On the other hand, the ethanolic extract obtained the highest phenolic content (981.84 mg GAE /100g). The antioxidant activity was evaluated through two spectrophotometric methods: DPPH• and FRAP. As a natural antioxidant, the use of ascorbic acid as a control allowed to measure the antioxidant activity of the bioactive compounds present in both extracts, when comparing them with a natural antioxidant. Although the antioxidant activity of ascorbic acid was superior in the two extracts, by the two spectrophotometric methods, these can be used as natural antioxidants, and it is through the ethanolic extract that we can take advantage of the antioxidant activity of the bioactive compounds of the seeds of Carica papaya Linn (416.87 μg/ml by DPPH• and 283.52 μg/ml by FRAP). With the obtained results we can conclude that the content of bioactive compounds is influenced by several factors, namely, the type of solvent extractor used. Although papaya seeds can be used as natural antioxidants, both in the food, pharmaceutical and cosmetic industries, it is necessary to deepen the investigations in order to evaluate their toxicity.
N/A

Made available in DSpace on 2015-05-14T13:21:47Z (GMT). No. of bitstreams: 1 arquivototal.pdf: 1669083 bytes, checksum: cbab7afab21f043f3d50f78626823f3f (MD5) Previous issue date: 2010-07-23
Coordenação de Aperfeiçoamento de Pessoal de Nível Superior - CAPES
The aim of this research was to study the chemical and physical properties of the papaya seeds of three different cultivars produced in Brazil: Sunrise Solo (Hawaii), Golden and Formosa. The chemical composition of the seeds and the extracted oils, physical chemical properties of oils and obtained biodiesel were analyzed to evaluate the industrial use of this disposable residue as renewable energy source. Golden and Solo Sunrise cultivars presented lower fruit weight (391.92 and 590.32 g) compared to Formosa cultivar (1318.16 g) but a higher seed yield of 2.54 and 1.43% respectively against 0.61% of Formosa. The chemical composition of the seeds shown high protein content (23.58 to 25.46 g/100 g), fibers (16.07 to 19.19 g/100 g) and minerals (5.12 to 6.48 g/100 g) among the three cultivars with emphasis on Fe and P (104.18 mg/g and 634.12 mg/g) for the cultivars Golden Sunrise Solo cultivars. The oil content was ± 28%. The major component were oleic acid (69.78%, 70.13% and 72.04%) and palmitic acid (18.95%, 18.89% and 18.20%) respectively for Golden, Sunrise Solo and Formosa cultivars. The acidity index (1.0, 0.98 and 1.03), iodine index (71.30, 70.26 and 70.45) and saponification index (190.2, 190.14 and 189.98) were found form the three papaya variety. Methyl transesterification of the Golden oil was carried out by 0.5% basic catalysis, 1:8 molar ratio, 45°C temperature and the product was identified by TLC, GC-MS, IR and 1H and 13C NMR techniques. The yield of the methyl esters (92.07%), was gravimetrically determined according to the stoichiometry of the reaction. The kinetics of acid catalyzed methyl transesterification using the same oil was evaluated by TLC and the yield of esterification was determined by reducing the acidity index of the reaction medium. The reaction medium used and results found were: 1:9 molar ratio and 80°C temperature the acidity index was reduced from 28 to 3.31 and 3.12 mg KOH/g sample, yielding of 88.18% and 88.86% respectively, at 1.0 and 1.4% catalyst after 8 hours of reaction. In a second stage of the reaction after 2 hours, the acidity index were 1.8 mg KOH / g (1.0% of catalyst) and 1.6 g KOH / g (1.4% of catalyst) with esterification maximum respectively 93.57 and 94.28%. The biodiesel obtained by basic catalysis showed 0.35 mg KOH / g acidity value, 0.88 g/cm3 density, 6.0 mm2 / s kinematic viscosity, 1°C cloud point, -1°C flow point and 62.83 cetane number. The power generation consumption, in L / h, were 0.5837 (B100), 0.5806 (B50), 0.5814 (B20), 0.6122 (B10), 0.6225 (B5) compared to 0.6981 (diesel), indicating its high calorific power. Thermal stability studies showed that the initial temperature of mass loss in TGA (155 ° C) match with the temperature variation of enthalpy by DSC curve and the initial oxidative event shown in non-isothermal curve PDSC indicating superior thermo-oxidative stability comparing to Soybean biodiesel where similar thermal events begin at a temperature of 100 º C. The oxidative stability (25 hours) was evaluated by Rancimat test. This result was confirmed by PDSC, where the variation of the baseline occurred at 54 minutes after reaching the temperature of the isotherm, at 110°C, and any other thermal event was observed during the entire period of the experiment (10 hours), indicating no oxidation reactions. The peroxide index was evaluated 58.4 meq / kg by using an oven (65 ºC) for a period of 25 days, showing a high stability if compared with the sunflower oil biodiesel, which according to the literature and in the same procedures presented a peroxide index of 300 meq / kg in seven days. The rheological study showed a Newtonian behavior and an absolute dynamic viscosity of 5.3 ± 9.1.10-5 m.Pa.s.
Este trabalho descreve as propriedades físicas das sementes de três cultivares de mamão produzidos no Brasil: Sunrise Solo (Havaí), Golden e Formosa, a composição química das sementes e dos seus respectivos óleos, as propriedades físico-químicas do óleo e do biodiesel dele obtido. Tudo isso, tem o objetivo de avaliar o aproveitamento industrial integral desse resíduo totalmente descartável , o qual é uma possível fonte renovável de energia. Os cultivares Golden e Sunrise Solo apresentaram menor peso de fruto (391,92 e 590,32 g) que o Formosa (1318,16 g); possuem, porém, um rendimento de sementes superior de respectivamente 2,54 e 1,43% contra 0,61 % da cultivar Formosa. A composição química das sementes em g/100 g apresentou um teor elevado de proteínas (23,58 a 25,46); fibras (16,07 a 19,19) e minerais (5,12 a 6,48) entre os três cultivares com destaque para os minerais Fe e P, cujos maiores teores foram, respectivamente, os de 104,18 mg/g e 634,12 mg/g para as cultivares Golden e Sunrise solo. O teor de óleo dos três cultivares foi o de ± 28% sendo majoritários os ácido graxos oléico com percentuais de 69,78; 70,13 e 72,04 e palmítico com percentuais de 18,95; 18,89 e 18,20, respectivamente para as cultivares Golden, Sunrise solo e Formosa. Na mesma ordem os índices de acidez : 1,0, 0.98 e 1,03, iodo: 71,30; 70,26 e 70,45 e saponificação: 190,2; 190,14 e 189,98. A transesterificação metílica do óleo do cultivar Golden ocorreu por catálise básica, na razão molar 1:8, temperatura de 45 °C e concentração de 0,5% de catalisador, sendo a conversão identificada pelas técnicas de: CCD, CG-MS, IR e RMN 1H e C13. A taxa percentual de conversão em ésteres metílicos (92,07) foi determinada por gravimetria, de acordo com estequiometria da reação. A cinética da reação de transesterificação metílica por catálise ácida no óleo de alta acidez do cultivar Golden foi avaliada por CCD e o percentual de esterificação da reação determinado pela redução do índice de acidez do meio reacional. Na razão molar de 1:9, temperatura de 80 °C, a acidez baixou de 28 para 3,31 e 3,12 mg de KOH/g de amostra com um percentual de esterificação de 88,18 e 88,86, respectivamente, nas concentrações de 1,0 e 1,4% de catalisador após 8 horas de reação. Numa segunda fase do processo reacional, após 2 horas, a acidez caiu para 1,8 mg de KOH/g (1,0% de catalisador) e 1,6 g de KOH/g (1,4% de catalisador), com esterificação máxima, respectivamente, de 93,57% e 94,28%. O biodiesel obtido por catálise básica apresentou índice de acidez (0,35 mg de KOH/g), densidade (0,88g/cm3), viscosidade cinemática (6,0 mm2/s), ponto de névoa (1 ºC), ponto de fluidez ( 1 ºC) e número de cetano (62,83). O consumo em gerador de energia em L/h mostrou os valores: 0,5837 (B100), 0,5806 (B50), 0,5814 (B20), 0,6122 (B10), 0,6225 (B5) contra 0,6981 (diesel). Os estudos de estabilidade térmica mostraram que a temperatura inicial de perda de massa na curva TG (155°C) coincidiu com o início da temperatura de variação da entalpia pela curva DSC e com o evento oxidativo inicial demonstrado na curva PDSC não isotérmica, demonstrando estabilidade termoxidativa superior ao biodiesel de soja, onde os eventos térmicos similares iniciam na temperatura de 100 ºC. No teste Rancimat, o biodiesel apresentou estabilidade oxidativa elevada (25 horas). Este fato foi confirmado pela técnica PDSC, durante a qual a mudança da linha de base ocorreu aos 54 minutos, atingida a temperatura da isoterma (110 ºC), não sendo registrado nenhum evento térmico (indicativo de reações de oxidação) por 10 horas. No método da estufa após um período de 25 dias a 65 ºC, o índice de peróxido máximo do biodiesel foi de 58,4 meq/Kg, uma estabilidade muito superior ao biodiesel de girassol, que segunda a literatura e nas mesmas condições analíticas apresentou um índice de peróxido de 300 meq/Kg em 7 dias. O estudo reológico demonstrou um comportamento newtoniano com uma viscosidade dinâmica absoluta de 5,3 ± 9,1. 10-5 m.Pa.s.

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior
The present study aimed to develop natural extracts from papaya seed (Carica papaya l.) and marcela (Achyrocline satureioides) to be used in chicken sausage, evaluating their antioxidant and anti-microbial activities. Firstly, the hydroethanolic extracts were elaborated and the phenolic compounds characterization and in vitro antioxidant activity were accomplished. Then, the papaya seeds and marcela extracts, in 0,5%, 1%, 1,5% and 0,5 and 0,75% respectively were used in the chicken sausage. The analyses accomplished were: centesimal composition (humidity, protein, ash and fat), pH, objective color, lipid oxidation, microbiological and sensory analyses. The results obtained in the extracts in vitro analyses showed that the marcela extract presents greater amount of total phenolic and higher antioxidant activity compared to the papaya seeds extract. The products centesimal composition complied with the Brazilian law. There was no interference of the extracts addition on the pH of the chicken sausage in relation to the control ones. Regarding color, during the storage period the T2, T3, T4 and T5 treatments presented higher values for h* in relation to control, indicating a color tendency from red to yellow. At the 42 days of storage of the refrigerated chicken sausage, the samples T3 (1,5% ESM) and T5 (0,75%EM) presented the lowest TBARS values, being respectively 0,485 and 0,448 mg malonaldehyde Kg-1 of sample, representing a lipid inhibition higher than control. The values obtained to Clostridium, Staphylococcus positive coagulase; Salmonella and coliforms at 45° were lower than the ones established by law. However the mesophylls aerobic and the psychotropic microorganisms were lower to 10-6 UFC. g-1 up to 21 days of storage, at 4 °C. The mean values of the scores attributed to color, smell, texture and global appearance did not present significant difference (p<0, 05) among the treatments. The acceptability index presented values higher than 70% for all attributes which is considered good. Then, it is possible to conclude that the addition of papaya seeds (1,5%) and marcela (0,75%) extracts can be used in the chicken sausage manufacture since they have presented antioxidant capacity and may lengthen this meat product s shelf life.
O presente estudo teve por objetivo desenvolver extratos naturais de sementes de mamão papaya (Carica papaya L.) e de marcela (Achyrocline satureioides) para aplicação em linguiça de frango, avaliando suas atividades antimicrobianas e antioxidantes. Primeiramente elaborou-se os extratos hidro-etanólicos e realizou-se a caracterização da composição de fenólicos e da atividade antioxidante in vitro. Após, os extratos de sementes de mamão e de marcela, nas concentrações de 0,5%, 1% e 1,5%, e 0,5% e 0,75%, respectivamente, foram aplicados em linguiça de frango. As análises realizadas nos embutidos foram: composição centesimal (umidade, proteínas, cinzas, gordura), pH, cor objetiva, oxidação lipídica, análises microbiológicas e sensorial. Os resultados obtidos nas análises in vitro dos extratos mostraram que o extrato de marcela apresenta maior quantidade de fenólicos totais e maior atividade antioxidante quando comparada ao extrato de sementes de mamão. A composição centesimal dos produtos estava de acordo com o exigido pela legislação brasileira. Não houve interferência da adição dos extratos sobre pH das linguiças de frango em relação ao controle. Em relação à cor, durante o período de armazenamento, os tratamentos T2, T3, T4 e T5 apresentaram valores superiores para o h* em relação ao controle, indicando uma tendência de cor vermelha para amarelo. Aos 42 dias de armazenamento das linguiças de frango refrigeradas, as amostras T3 (1,5%ESM) e T5 (0,75%EM) apresentaram os menores valores de TBARS, sendo respectivamente 0,485 e 0,448 mg malonaldeído.Kg-1 de amostra, representando uma inibição lipídica superior ao controle. Os valores obtidos para Clostridium, Staphylococcus coagulase positiva; Samonella e coliformes a 45 ºC encontravam-se com valores inferiores aos limites estabelecidos pela legislação. Já a contagem de microrganismos aeróbios mesófilos e psicrotróficos foram inferiores a 10-6 UFC. g-1 até os 21 dias de armazenamento, a 4 ºC. Os valores médios das notas atribuídas para os atributos cor, odor, sabor, textura e aparência global não apresentaram diferença significativa (p<0,05) entre os tratamentos. O índice de aceitabilidade apresentou valores superiores a 70% para todos atributos avaliados, sendo considerado com boa repercussão. Portanto, conclui-se que a adição dos extratos de sementes de mamão (1,5%) e de marcela (0,75%), podem ser utilizadas na elaboração de linguiça de frango, pois apresentaram capacidades antioxidantes, podendo aumentar a vida de prateleira desse produto cárneo.

Experiments were carried out with the objectives of developing propagation protocols for Uapaca kirkiana and Pappea capensis tree species of southern Africa, and evaluating the graft compatibility within U. Kirkiana tree clones, provenances and species. Reverse phase high performance liquid chromatography (RP-HPLC), Folin-Ciocalteau reagent, fluorescence microscopy and callus fusion methodologies were used to diagnose graft compatibility. Results indicated that U. Kirkiana culture asepsis was achieved with 0.1% w/v mercuric chloride HgCl2) and using pre-conditioned grafted trees. Sodium hypochlorite (NaOCl) improved P. Capensis seed asepsis and germination, and discarding floating seeds improved germination. Murashige and Skoog (MS) medium with 2.0 mg l-1 benzylaminopurine (BAP) and 0.3 mg l-1 casein hydrolysate (CH) was superior in shoot multiplication and 0.5 mg l-1 indole-3-butyric acid (IBA) for rooting of P. Capensis microshoots. For somatic embryogenesis, three quarter strength MS medium with 0.05 mg l-1 thidiazuron (TDZ) and 0.3 mg l-1 CH, or 0.2 mg l-1 BAP with 0.3 mg l-1 CH, were effective in germination of P. Capensis somatic embryos. For U. Kirkiana lateral shoot explants, shoot multiplication was superior on three quarter strength MS medium with 0.1 mg l-1 BAP and 0.3 mg l-1 CH. Rooting of micro-cuttings (36%) was achieved on ½ MS with 2.5 mg l-1 IBA. RP-HPLC, fluorescence microscopy and callus fusion studies showed that phenolic compounds play a major role in U. Kirkiana graft incompatibility. Less graft compatible combinations showed an increase in phenol deposits above the union and graft incompatibility was more pronounced above the union than below the union. Proliferation of parenchymatous tissues was better below the union than above the union. Fluorescence microscopy showed presence of flavonoids and polymers above the union of less graft compatible combinations. The chromatograms showed that ferulic acid was abundant and responsible for wood discolouration. The chromatograms also isolated ρara-coumaric acids which were predominant above the union of the less compatible combinations. Therefore, ρara-coumaric acids, flavonoids and polymers were implicated in graft incompatibility of U. kirkiana trees. Copyright
Thesis (PhD)--University of Pretoria, 2008.
Plant Production and Soil Science
unrestricted

碩士
國立臺灣大學
園藝學研究所
97
Papaya (Carica papaya L.), an important economical fruit crop in Taiwan, can be propagated by seeds, grafting, cutting and tissue culture, but mainly from seeds. Therefore, the germinability of seeds are vital to the industry. The characteristics of papaya seed germination are slow and asynchronous. Numerous researchers had proposed different kinds of methods to improve papaya seed germination, but the efficiency is significant different and contradictory depends upon cultivars and environment. The problem of poor and asynchronous germination of papaya seeds still exists. Thus, it is important for papaya nursery industry to look for effective methods to increase germination. Fresh papaya seeds harvested from‘Tainung no. 2’fruits at yellow-ripen stage are dormant after removing the sacrotesta and washing. Scarification and H2SO4 treatment have no effect on improving germination. The germination percentage can be achieved to 65% after removing the testa and tegmen, suggested that testa is the main physical constraint to radicle protrusion, instead of limiting the water and air permeability of seed. Fresh papaya seeds were after-ripened at 20℃ for 4 weeks and the water content of seeds decreased to 5.31~12.24%, but couldn’t germinate as well. Papaya seeds can improve germination by treated GA3 and GA4+7 solutions, and the most effective treatment is GA4+7 100 mg/L, implies that the main bioactive GAs for papaya seed germination may be GA4 and/or GA7. Seeds that were after-ripened for 14 days was soaked in 100 mg/L GA4+7 for 24 hours and incubated in Paclobutrazol and Ancymidol solutions. The inhibitory effects of these two inhibitors enhance as the concentration increases, demonstrates indirectly that the ability of papaya seeds to synthesize GAs continuously is quite important. Papaya seeds can’t germinate without adding GAs, implies that the embryo of papaya seed is weak. In addition to the weak embryo and the mechanical constraint of testa, papaya seeds are belonged to physiological dormancy. Although GAs treatment can improve the germination and synchronicity, it’s improper to be put in use due to the 30% abnormal seedlings occurrence. Papaya seed dormancy can be alleviated by hot water immersion treatment. 34, 36, 38, 40℃ hot water immersion treatment can enhance the germination of after-ripened seeds imbibed for 1 day. The best condition is 36℃ treated for 5 hours, and the germination percentage is 58.67%. The efficiency will be better as the imbibition duration before hot water immersion treatment increases to more than 5 days, the germination can be increased to 74%, but there’s no significant difference when the imbibition duration is more than 5 days. Compare the effects of different heat shock treatment, the positive improvement of hot water immersion (36℃, 5h) is better than hot air treatment (36℃, 5h), and after-ripened seeds are more sensitive to heat response than fresh seeds. Seedlings after heat shock treatment are healthy, have no side effects, unlike GAs treatment which will produce abnormal seedlings, and the method can be regarded as a dormancy breaking technology in papaya seedling propagation. In order to figure out the mechanism of heat shock treatment to improve papaya seed germination, we incubated seeds in Paclobutrazol and Ancymidol solutions after 36℃ hot water-immersion treated for 5 hours. 10 μM Ancymidol completely reverse the promoting effect of hot water immersion treatment. Counteracting effects of 10~250μM Paclobutrazol enhance as the concentration increases, implies that hot water immersion treatment should improve papaya seed germination via GAs de novo synthesis. According to the results of endogenous GA1, GA3, GA4, GA7, GA20 and ABA analysis, the GA4 content of hot water immersion treated seeds increases 1.14 times and enhances continuously as incubation time increases, but the heat treatment has no significant effect on endogenous GA3 content, implies that the bioactive form of papaya seed germination is GA4. The ABA contents of papaya seeds is not detected, suggested that ABA may not be the main factor influencing papaya seed dormancy and germination. This is conformed that 10~100 μM Fluridone treatment has no effect on improving germination. In conclusion, we proposed a hypothesis of mechanism how heat shock treatment can improve papaya seed germination. Seeds can restore physiological metabolism during the imbibition period. After heat stimulated, seeds were directly or indirectly induced increasing the rate of endogenous GAs synthesis or inhibiting the rate of GAs breakdown, and the GA4 content increases. Through GA signal transduction to start the degradation and mobilization of seed reserves and weaken the physical constraint of the cell wall of endosperm cap and testa, and then seeds can germinate successfully.

碩士
國立臺灣大學
園藝學研究所
94
A few seed germination internal factors such as influence of testa and tegmen, moisture content after imbibitions, maturation of fruits and seed size and weight and external environment such as influences of alternating temperature and light were investigate to sought for problem of papaya germination. The testa and tegmen presented markedly delay in germination as a physical resistance to radicle emergence. Tegmen might play a role in seed protection and germination because removal tegmen might cause abnormal seed. Moisture content of dried intact seeds and seedcoat(including testa and tegmen) increase linearly after imbibitions, and reach a plateau in short time. While moisture content of endosperm include embryo were lower but shown water absorption 3 phase. So, the moisture content of endosperm include embryo was the critical point to papaya germination and endosperm play a role in blocking water absorption. Fruit maturation was significant effect in papaya seed germination and germination rate. Seed size of papaya might not significantly effect on germination, while the heavier seed weight were showed better germination. Papaya germinated at alternating temperature 35/25℃ and 35/20℃(12h/12h) were higher than constant 35℃in 12 h photoperiod. Compared the germination of papaya at constant temperature 35℃, 30℃ and 25℃ and alternating temperatures of 35/30℃, 35/25℃, 35/20℃, 30/25℃ and 30/20℃ in darkness, alternating temperature regimes provided higher germination than constant temperatures. Germination percentage at constant temperatures 35℃ and 25℃ was able to increase progressively after transferring to alternating temperature. Continuous 24h light treatment inhibited papaya germination. Germination percentage was not significant difference between darkness or photoperiod 8h and 12h at 30℃ constant temperature. A factorial experiment of temperature regimes (outdoor temperature, fluctuating temperature, alternating temperature and constant temperature); light treatments (with 12 hours light or darkness) and GA3 treatments (with or without pretreated GA3 400mg/L 3 hours). The interaction between temperature, light and gibberellins treatment did not show significant effect on papaya germination, but the temperature had the significant effect on papaya germination.

博士
臺灣大學
農業化學研究所
98
Lectins are carbohydrate recognizing proteins which prevanlently exist in most, if not all, living organisms. For a long time, plant lectins were commonly perceived as ‘antinutrients’, mainly due to their adverse effects of causing non-pathogenic food-borne poisoning when not properly cooked before consumption. However, recent studies exhibited that several lectins have been found to possess anticancer properties and showed promising potential as anticancer agents.This study provides a new strategy to detect the specific carbohydrate binding capability of lectins. A sugar-polymer based enzyme-linked adsorbent assay was established by applying different monosaccharide-polyacrylamide conjugates, including D-mannose, N-acetyl-D-glucosamine, N-acetyl-D-galactosamine and L-fucose, as capturing agents for screening lectins in biological samples. Four model lectins, concanavalin A (Con A), wheat germ agglutinin (WGA), soybean agglutinin (SBA) and Ulex europaeus agglutinin I (UEA I), were employed as the comparative lectins against each corresponding sugar. The results suggested that this assay is more sensitive than the conventional hemagglutinating methods. On the screening of specific carbohydrate binding capability from 29 plant materials, a specific N-acetyl-D-galactosamine (GalNAc) binding capability was observed in the crude extracts of papaya (Carica papaya) seeds. The GalNAc specific binding protein was subsequently isolated and designated as CPL (C. papaya lectin). Purification of the lectin involved ammonium sulfate fractionation and DEAE anion exchange and repeated gel filtration chromatography. Inhibition of CPL causing hemagglutination on human erythrocytes showed that the lectin shows specificity to GalNAc and lactose. Surface plasmon resonance further revealed that the lectin possesses high specificity toward GalNAc with a dissociation constant of 5.5 × 10^-9 M. The lectin is composed of 38- and 40- kDa subunits with molecular mass of ~804 kDa estimated by size-exclusion high-performance liquid chromatography. Incubation of CPL with Jurkat T cells showed significant induction of IL-2 cytokine, which suggests that CPL has potent immunomodulatory effects on immune cells.

碩士
臺灣大學
農業化學研究所
98
Based on our previous study, a novel N-acetylgalactosamine binding protein in papaya seed (Carica papaya lecin, CPL) was discovered by GalNAc-polyacrylamide based enzyme-linked adsorbent assay (Food Chem. 2009, 113: 1218-1225). Papaya seeds with different maturity were collected, and the extracts of seeds in green papaya fruits shows relatively high activity. Further purification of CPL was conducted as following: freeze-dried papaya seeds were ground into powder, extracted with 20 folds (w/v) of 50 mM phosphate buffered saline (pH 7.4) at 4°C overnight, centrifuged and collected the supernatant, added ammonium sulfate to 70% saturation. Precipitations were resuspended and dialyzed against PBS, then fractionated by a 50 kDa MWCO ultrafiltration. The retentate was further purified by HiTrap CM FF ion exchange chromatography and Superdex 200 GL gel filtration chromatography. SDS-PAGE and HPLC gel filtration indicated that CPL is a polymer with a molecular mass of 804 ± 30 kDa and composed of two different subunits of 38 and 40 kDa associated by non-covalent bonds. It was heat stable until up to 70oC for 30 min and showed optimum sugar-binding activity from pH 6.0 to 8.0. Also, CPL did not require Ca2+, Mg2+, Mn2+, Zn2+for its activity. Of various sugars tested, the lectin was best inhibited by GalNAc. CPL agglutinated all trypsinized human RBC types, with a slight preference for the A blood group which immunodeterminant is GalNAc. Based on the specificity, we performed an alternative ripid, simple purification method via GalNAc-Sepharose 6B affinity chromatography column, and gave the lectin with a 6000-fold purification as compared to the crude extract.

碩士
國立臺灣大學
農藝學系
86
Due to their dormancy character and low germination rate, germination test of papaya seeds is not only time-consuming, but may under-estimate their true viability. In addition, papaya seed is not able to tolerate both dry and low temperature conditions, hence making preservation of the germplasm difficult. The object of this thesis is to improve the electrical conductivity and TTC test methods that are alternative ways to determine the viability of papaya seed rapidly, and to investigate in more details the storage behavior of papaya seeds under freeze temperature and low seed moisture content. The moisture content of papaya seeds, in between 3.70% to 11.64%, did not interfere with the results of both the electrical conductivity and TTC test methods. As high as 70% of the dead papaya seeds that were obtained by accelerated aging stained positively by TTC test. These data were explained by the action of the hard inner seed coat. A new TTC test procedure was developed. It was shown that 18-22 hrs soaking of papaya seeds at 25℃, followed by another 14-22 hrs imbibition of the embryos, distinguished the viable from the accelerated-age seeds. The new procedure stained the viable embryo into red or orange color, while the dead embryo pink or white. Those imbibition schemes of longer or shorter seed soaking time under- or over- estimated the germination percentage of the samples respectively. The new procedure was examined with 54 samples of various sources. It was shown that the germination percentages were always higher than 85% for those samples with viability higher than 90% as estimated by the new TTC test method. During imbibition, the electrolyte-leaching rate of papaya seed rapidly decreased immediately after soaking. However, only those of aged seed revealed a second phase, which was 24th - 48th hr of imbibition, of high leaching rate. This second phase was founded to be more stable at 25℃, and could be revealed distinctly by sample size of 50 seeds and soaking media of 50 ml de-ionized water. The correlation between germination percentage and the amount of leachate conductivity of the second phase as tested by the above conditions can be improved further, if those samples which containing empty seeds were eliminated. Papaya seeds were sensitive to low temperature and they were better kept under temperature over 0℃ with moderate moisture contents. The seed of ''Tainung no.2'', with moisture content 3.7%-11.64%, lost viability after storing at -25℃ for 6 months, while unexpectedly, 70% of the seeds of ''Sunrise'', with moisture content 3.98%-9.23%, were still viable after storing at -25℃ for 7.5 months. The new TTC test, but not the electrical conductivity test, proved reliable in estimating the viability of those papaya seeds that was freeze-stored.

碩士
靜宜大學
食品營養研究所
98
A chitinase was purified from lyophilized powder of mature papaya seeds through successive steps of buffer extraction, ammonium sulfate precipitation, Sephacryl S-100 HR gel filtration, Polybuffer exchanger PBE 94 chromatofocusing and Sephacryl S-100 HR gel filtration. By these steps, the purity of the enzyme was increased 152 fold and the recovery of enzyme activity was 17.5％. The purified chitinase catalyzed the hydrolysis of both chitin and chitosan. The optimum pH for CM-chitin hydrolysis was 4.5, the optimum temperature was 50℃, the Km was 1.88 mg/mL and the Vmax was 1.08 μmol GlcNAc/min/mg, whereas the optimum pH for AE-chitosan hydrolysis was 6, the optimum temperature was 50℃, the Km was 1.20 mg/mL and Vmax was 0.130 μmol GlcN/min/mg. The purified chitinase was thermally stable after holding at 30-50℃ for 60 min. However, at temperatures higher than 50℃, the enzyme activity decreased significantly. The initial heat inactivation of the enzyme followed first-order reaction kinetics. The half-life values of thermal inactivation for the enzyme varied from 51.5 to 3.4 min at 60-70℃. The isoelectric point of the enzyme was pH 4.6, as estimated by isoelectric focusing electrophoresis and zymogram staining. The molecular mass of the enzyme was 31.6 kDa, as estimated by gel filtration. This value was close to that estimated by SDS-PAGE (33.1 kDa), indicating the purified chitinase was a monomeric enzyme. The purified chitinase was deemed to be a class III chitinase because it was recognized by a polyclonal antibody raised against 30 kDa chitinase from jelly fig achene, as examined by Western immunoblotting. The purified enzyme was not fully homogeneous, there was some contamination with trace amounts of a minor isoform of chitinase (28.3 kDa), as analyzed by SDS-PAGE and chitinase activity staining. The minor isoform was not recognized by anti- (jelly fig chitinase) antibody. Chemical modification agents, including diethyl pyrocarbonate (2.5 mM), Woodward’s reagent K (50 mM), N-bromosuccinimide (5 mM) and N-acetylimidazole (2.5 mM) significantly inhibited the activity of the purified chitinase, implying that imidazole groups from histidine, carboxyl groups from aspartic acid and glutamic acid, indole groups from tryptophan and phenolic groups from tyrosine were located at or near the active site of the enzyme. The purified chitinase catalyzed the hydrolysis of chitin, chitosan and their derivatives. However, the enzyme had greater activity toward chitin than toward chitosan. The major product of ethylene glycol chitin hydrolysis catalyzed by the enzyme was chitin oligomer with chain length of 4.

碩士
國立臺灣大學
農藝學系
82
Papaya is dicotyledonal plant with endosperm in their seeds. After removing the sarcotesta, many single-ridges and one double- ridge could be found on surface. The weight of seedcoat is 47% of the seed and is composed of the testa and tegmen. The outer layer of endosperm is aleurone layer. Embryo is axile spatulate type. The width of seed was 4mm and the thickness of seedcoat was 1mm. Seedcoat could be thought as mechanical boundary for germination. The stages of germination could be classified as imbibition, seedcoat cracking, radicle emergence, elongation of secondary roots and hypocotyl, and the stage that cotyledons separated from aleurone layer. Changes of moisture content during germination could be decided by mositure content of embryo and endosperm. The initial moisture cntent of seed was 6.2 %, and that of embryo and endosperm was 3.9%, 25.8% at 24h after incubation, 43.7% at seed- coat cracking stage (3-6 days after incubation), 58.8% at radicle emergence (5-8 days after incubation). Rolled paper was suggested to be the best substratum. Optimum temperatures for germination test were found to be 30 degree C, 30/25 degree C and 27 degree C. Under these conditions, the first and the final count of germination were 8th and 17th day after incubation, respectively. Percentage germination was not affected by light treatment. A normal seedling could be defined by follow characteristics : one primary root and four secondary roots, no spiralle, constrict and necroses observed on the hypocotyl and cotyledons, the seedcoat and aleurone layer had left cotyledons during germination. The treatment of KNO3 (1M) could promote the germination rate of papaya seed, but was less effective than that of GA3(500 ppm). Leaching was not effective in promoting germination, but it could promote the germination rate of the seeds of low dormancy.

碩士
國立臺灣大學
園藝學研究所
99
Papaya (Carica papaya L.) is one of the important economical fruit crops in Taiwan, and it can be propagated by seeds, grafting, cutting and tissue culture. However, papaya is propagated mainly by seeds. Nevertheless, freshly harvested papaya seeds germinate poorly and unevenly. Even though numerous researchers had reported many kinds of treatments to improve the germination, results were different due to cultivars and environmental factors. Gibberellins and heat treatments have been reported to promote the germination of papaya seeds. However, the mechanism of how heat treatment breaks the dormancy of papaya seeds is not clear. Therefore, it is an important topic to understand how heat treatment induces the germination of papaya seeds. In this research, I use the method of proteomic analysis to investigate the possible mechanism of heat treatment on the dormancy breakage of papaya seeds. Unfortunately, there are no related protocols have been reported for protein preparation from papaya seed for proteomics analysis. Therefore, in this study I established the optimized protocol of protein extraction for sample preparation of two-dimensional electrophoresis. The protein extraction mixture for 5 day-imbibed papaya seeds was a Tris-HCl base buffer. The extracted solution was then precipitated by 10% TCA/Acetone (TCA-A) or extracted by phenol followed by 0.1M ammonium acetate-methanol precipitation (P-AA-M). The results showed that the method of P-AA-M could increase at least 5 bands than TCA-A in SDS-PAGE analysis. Two of the bands were assessed via LC-MS/MS and the proteins identified included “unknown proteins”, with the highest ratio (34%), “energy metabolic proteins”, the minor category (20%), and “cell cycle proteins”( 14%). P-AA-M could reduce the problems of streaking and smearing in 2D gel analysis, enhance the efficiency of protein separation, and therefore, increase the number of protein spots identified. Application of 36℃ hot water immersion for 5 hours to 5 day-imbibed papaya seeds would induce the expression of proteins related to calcium signal transduction (CBL and CDPK), small heat shock protein (17.5 sHSP), energy metabolism (G3PDH and ATP synthase epsilon chain), and transcription (GR-RBP). It is assumed that after the heat treatment, the papaya seeds would be released from dormancy by a series of physiological events including signal transduction and energy metabolism, increase of the embryo growth potential, overcome the physical constraint of testa, and then germination. According to the results, the possible mechanism of heat treatment on dormancy breakage of papaya seeds would be through GA, but not brassinoids, biosynthesis, since brassinzole, a brassinoids synthesis inhibitor, was unable to inhibit the germination of the seeds treated with the hot heat immersion Even though heat treatment could induce the germination of papaya seeds, the effects would be various depending on degree of the dormancy. Hot water immersion treatment with 34, 36, or 38℃ could enhance the germination of after-ripened seeds. The longer the after-ripening time applied, the better the efficacy of dormancy releasing. Moreover, the germination capability of papaya seeds acquired from heat treatment could be preserved up to 80% if the seeds were prepared by fast drying method (36℃ forced air for 6 hours), but could not be preserved by slow drying method.

碩士
中興大學
園藝學系所
99
When waterlogged, plants suffer two injuries: accumulation of toxic metabolites and lack of energy caused by the shortage of oxygen. Papaya (Carica papaya L.) is severely waterlogging intolerant and was used to study two methods of sorting seeds for waterlogging tolerance. The first method exposed papaya seeds to ethanol, a toxic metabolic that is the product of anaerobic respiration. The second was to establish an accelerated method of sorting by exposing papaya seeds and leaves to hypoxic (lack of oxygen) stress which was simulated by nitrogen gas. Results showed that the ethanol treatment of ‘TN2’ papaya seeds reduced their germination percentage and extended their germination time in direct correlation to the amount of ethanol and duration of the treatment. The most significant results were seen at 1.0 mL/ 4L ethanol for 3 days in which 66% of the seeds did not germinate and few that did required 15 days to germinate, a process that only takes 10 days under normal conditions. This suggested that the physiological function of seed was obstructed by ethanol. Twelve cultivar seeds were then sorted using this treatment (1.0 mL/4L ethanol for 3 days). The mean germination time of the seeds was increased significantly but the changes in germination were non-significant when compared to the control group. This may indicate that germination of the seeds, which were pretreated with ethanol, might also have been affected by other factors, but this requires further study to confirm. After the ethanol-germination tests, the resulting seedlings underwent waterlogging experiments in which the seedlings were immersed in water for 2 days. The appearance of the seedlings and chlorophyll fluorescence of the leaves did not change significantly. Nitrogen gas created an oxygen deficient (oxygen concentration was below 5%) condition to which ‘TN2’ papaya seeds were exposed at 0, 5, 10, 15, and 20 days after being sown. The germination decreased significantly while the mean germination time increased, especially in the seeds that were exposed to nitrogen on the 10th day after being sown. This is the point in the germination process when the seeds are the most sensitive to oxygen deficiency. Therefore, this treatment can be used as an indicator for sorting. In the nitrogen gas treatment of leaves, the membrane stability index and chlorophyll fluorescence of the leaves of adult papaya plants decreased significantly after 36 hours, more so for mature leaves than for new and older leaves. Changes in the membrane stability index and chlorophyll fluorescence showed a positive correlation; therefore, chlorophyll fluorescence test were used to quickly sort 17 cultivars of papaya for possible waterlogging tolerance. ‘PPI x ML-F2’ was found to maintain the highest chlorophyll fluorescence and ‘Exotica’ showed the lowest chlorophyll fluorescence.

碩士
環球科技大學
生物技術研究所
102
Papaya (Carica papaya) is a species in genus Carica of the plant family, is now ranked third of the total tropical fruit production. The purpose of this study was to investigate the feasibility of papaya seeds as the alternatives ingredient for soy sauce. In this study, we using the same condition of the experiment design which compared 3 different ingredients of soy sauce: soybean (SB), papaya seed (PS), and soybean with papaya seed (SBPS). The preparation to make the soy sauce was carried out by two stages: koji-fermentation and moromi-fermentation. The koji was prepared by mixing Aspergillus oryzae with the steamed ingredient (SB, PS or SBPS) and incubated for seven days. Moromi which consists of the koji and the salt brine was fermented for 90 days at 37°C. After the fermentation, the moromi was extruded, filtered, pasteurized to produce the final product soy sauce. The soybeans crude protein is about 39% which papaya seeds crude protein is only about 26% (2/3 of soy beans crude protein). The weight of each raw material was calculated from the crude protein. The crude protein of each sample should be equal to 39%. Then these raw materials been inoculated with Aspergillus oryzae and fermented for 7 days. Koji fermentation for 7 days, and total nitrogen of SB, SBPS and PS is 6.1%, 4.9% and 4% at the end of fermentation process. After 7 days fermentation, the protease enzyme activity (at pH 6.0) in SB koji reached 944 U/g dry koji which the highest of the others, while SBPS and PS koji reached only about 322.6 and 579 U/g dry koji. Higher protease activity can affect the hydrolysis level of protein and can promote the produce of protein decomposition products. After 90 days of moromi fermentation, the soy sauce of SB became darker and the other samples have a lighter color than SB. Only SB soy sauce can be classified as Grade A of soy sauce (total nitrogen content higher than 1.4 g/100 mL and amino nitrogen content higher than 0.56 g/100 mL). SBPS and PS soy sauce only contains about 0.26 g/100 mL and 0.07 g/100 mL for the total nitrogen, 0.07 g/100 mL and 0.04 g/100 mL for the amino nitrogen content.

碩士
國立臺灣大學
園藝暨景觀學系
102
Seeds are the main propagules for papaya (Carica papaya L.), which exhibit physiological dormancy. Although the germination percentage was enhanced and the germination time was reduced via heat shock treatment, the underlying mechanism is still unrevealed. The germination ratio of 2 week-dry papaya seeds extraced from fully ripe ‘Tainung no. 2’ papaya was only 0-3% after imbibition in water for 16 days in dark at 26℃; however, the ratio increased to 44±4%% and 67±5%, respectively, after immersion in water with 34℃ for 3 h and 36℃ for 5 h. The content of GA4, a bioactive GA, in the seeds treated with 36℃ water immersion for 5 h was 103.8 ng&;#8226;g-1 DW, which was higher when compared with 45.0 ng&;#8226;g-1 DW of control. However, the stimulating effect of germination was not offset in papaya seeds transferred to 10-2 ~ 10-3 M prohexadione, a GA biosynthetic inhibitor, between 24 h before and during the heat shock procedure. By ultilizing RT-PCR strategy, two cDNA clones of GA 20-oxidase (GA20ox) and GA 2-oxidase (GA2ox), two key enzymes of GA metabolic pathway in planta, were obtained from the tissures of papaya seeds. The full-length GA20ox cDNA was 1705 bp, deriving 385 amino acids, with binding sites for gibberellin substrate and Fe2+ as well as four conserved amino acids and was named CpGA20ox. The GA2ox cDNA clone had 1362 bp, deriving 335 amino acids, with three conservative domains and five conserved amino acids, including a gibberellin substrate, and was named CpGA2ox, a Classes I GA 2-oxi gene,. CpGA20ox was not up-regulated during the 5 h of 36℃ heat shock treatment, but the gene expression was induced to 307 and 2.4 times at the first and the fourth day at 26℃, respectively, after the heat treatment. On the other hand, CpGA2ox expression of the heat-shocked papaya seeds was suppressed to 0.38 fold on the fourth day after heat shock. Since the key enzymes in GA biosynthetic pathway were multi-gene family, the data obtained in this research were unable to clearly depicted the mechanism of the germination enhancement in papaya seeds via heat shock treatment.

碩士
國立高雄海洋科技大學
水產食品科學研究所
101
Papaya (Carican papaya) seeds are the wastes that come from being eaten fresh or processed papaya. Proximate compositon in dry weight showed that papaya seed contains 33.1% of lipid and 26.2% of protein. If the wastes can be further developed into useful products, it can increase the added-value of agricultural and enchance economic efficiency. Response surface methodology, (RSM) was emplooyed to evaluate the optimized parameter for maximum protein extraction from papaya seeds.The result was as follows: NaCl solution concentration, 1.5 M; solute:NaCl solution ratio=1:40 (w/v); extraction time, 2 h. The recovery of 94.8% of papaya seed protein was obtained and isoelectric point was determined as being pH 4.8. The functional properties of commercial soy protein isolate (SPI) and papaya protein were determined for comparison. The papaya seed protein had a poorer solubility and water-holding capacity at the pH 7.0 and 25C than SPI, but oil holding capacities, emulsifying activity, foaming capacity and foaming stability of papaya seed protein were higher than those of SPI. Furthermore, emulsifying properties of PSP as a function of concentration (0.5%、1.0%、1.5%及2.0%) and pH (3.0、4.0、5.0及6.0) were studied and their emulsifying capacity, emulsion viscosity, droplet distribution and emulsion type were compared. The results demonstrated that PSP had the higher emulsifying capacity at pH 3.0 than that at pH 5.0. In the range of PSP concentration of 0.5%2%, the emulsifying capacity was highest with 1.0% PSP. Referring to viscosity and stability of emulsion, they exhibited to be excellent at pH 3.0, regardless of concentration. While at pH 5.0,the emulsion had very poor viscosity and bad stability. Droplet distribution of emulsion is better with more stable emulsion, giving an emulsion with droplet size of less than 100 μm. Replacement of PSP for egg yolks was prepared for French salad dressing and was compared with conventional one (adding 0.6% egg yolks) on the sensory evaluations, viscosity, stability and microbial growths. The result showed that among the all groups, the added with 1.0% PSP had poor performance.While added with 1.0% PSP group had slightly high acceptability on mouthfeel, flavor and sour than traditional salad dressing. However, the color acceptance for added with 1.0% PSP group than was much worse than that for traditional one.

碩士
國立中興大學
園藝學系所
97
The fresh and dry seed weight of papaya were increased following the maturity of fruit. The weight of sacrotesta, testa and tegmen was over 1/3 of total seed weight. Using sugar density for grading floating and sinking seeds showed that both kind of fresh seeds had low percentage in germination of 7.4-9.2%. After 5 hours of incubating at 37℃ hot water, could increase percentage significantly. Under 1.05 sugar density, the percent germination of floating and sinking seeds were 22.3 and 41.9%, respectively. The germination percentage of different maturity fruit were increased afterripening. Seed moisture content of 8 and 12% had no significant difference in germination percentage but it was significantly improved by increasing seed storage time. Seeds dried by 35℃ with 3 months storage had 45% germination which is significant higher than 35% of seed dried by 26℃. The optimum temperation in seed germination is 30℃, but using alternate germination temperature of 25/30℃ and 25/35℃ had higher germination of 88.8 and 96.6%, respectively. Seeds incubating at 37℃ and using GA3, KNO3 could improve seed germination percentage. Seeds treated the solution of GA3 and KNO3 had higher germination percentage than that treated GA3 and KNO3, respectively. Solid and liquid priming could shorten the mean days to germination. Measuring EC in leakage and TTC method could be used for quick viability test.

博士
國立中興大學
園藝學系所
101
Studies on effects of natural type S-ABA on seed germination and seedling growth of papaya (Carica papaya L. cv. Tainung No. 2) were evaluated for clarify function of S-ABA. Seed germination was investigated by soaking seeds into distilled water and various concentrations of S-ABA. Percentage of seed germination in filter paper and growth medium experiments were increased by S-ABA treatments. These treatments significantly increased germination percentages (88 and 91 %, respectively) compared to control (70 and 76 %, respectively). Enhancement of germination velocity index (GVI) and seedling vigor index (VI) were obtained from treatment of S-ABA 0.01 and 0.1 ppm. While, high concentration of S-ABA (10 ppm) reduced seed germination, and resulted in abnormal germinants. However, S-ABA 1 ppm had no effect on seed germination. Young seedling stage, seedlings were treated by 2 methods of S-ABA application, foliar spray and medium supply. Results showed that S-ABA treatments increased seedling growth, especially number of leaves and roots, and fresh weight of shoot in both two application methods. Fresh weight of roots was increased by S-ABA only in foliar spray method while medium supply of S-ABA resulted in roots were small and fine. The investigation of anatomical characteristics in leaf, stem and root was supported this study. Number of palisade cell in leaf of treated seedlings was significantly increased compared to control ones. In addition, length of these cells in treated seedlings was increased but width of cell was reduced, led to leaves were dark green compared to untreated seedlings. S-ABA treatments could improve number of cell in stem and root and also increase vascular system in stem and root. Moreover, increasing in layer number of pith in stem was found from S-ABA treatments as well. Further, arrangement of cell in all three parts was more compact compared to untreated seedlings. Effects of S-ABA on nutrient and total sugar content were evaluated. S-ABA could significantly improve N and K content in all parts of seedlings, P and total soluble sugar content was increased trendily in some part of seedling. For dry weight of nutrients in whole plant, S-ABA could improve N, P, K and total soluble sugar in both two treatments of foliar spray and medium supply. Further, in 6 weeks after treatment, medium supply of S-ABA became to be more positive effect than foliar spray plants. It was found in this study that S-ABA belongs to growth promoter in tropical fruit tree, papaya. The concentration in 0.1 ppm was more effect on seedling growth and nutrient accumulation. The foliar spray of S-ABA had positive effect on leaf growth, thereafter indirectly improved stem and root growth. Moreover, the introducing of medium supply method directly enhanced root growth.

碩士
國立臺灣大學
農業化學研究所
100
Lectins are defined as sugar-binding and cell-agglutinating proteins of non-immune origin. They are ubiquitous in nature, being found in all kinds of organisms, from viruses to humans. Recently, our group isolated an N-Acetyl-D-galactosamine (GalNAc)-specific lectin from the seeds of papaya (Carica papaya), named CPL. In this study, an effective affinity chromatographic method for CPL isolation and purification was established, based on the specificity of CPL. By using two steps affinity chromatography through lactose-Sepharose 6B and GalNAc-Sepharose 6B resins, with sequential elution by lactose and GalNAc solution. The hemagglutination activity of GalNAc-elution fractions (LpGLac-GalNAc) was 1461 fold higher than original papaya seed crude extracts. Through the gel filtration chromatography, the results showed that the purified CPL was appeared in LpGLac-GalNAc; also, this purification protocol obtained higher purity and higher protein yield of CPL than previous research (Wang et al., 2011). At the same time, we discovered a lactose desorbed protein, designed as LpGLac, in the process of affinity purification. The results of SDS-PAGE, hemagglutination assay and ELISA assay indicated that LpGLac was recognized by CPL and might compete with lactose for carbohydrate binding domain on CPL, which resulted in desorbing from affinity column by lactose solution. In addition, we investigated the immunomodulatory effects of purified protein on murine macrophages RAW 264.7 cell line in vitro. Incubation of LpGLac and LpGLac-GalNAc with RAW cells showed significant induction of pro-inflammatory cytokines, IL-6 and TNF-α, as well as nitric oxide, which suggests that both of purified proteins have potent immunomodulatory effects on immune cells.

博士
國立中興大學
園藝學系所
100
Smoke-water is a chemical extract used to stimulate the germination of seed, seedling growth and inhibit disease spreading of many plant species under cultivation. The present study was initiated to understanding the effect of smoke-water on germination, growth, defense to pathogenic fungi and increased postharvest quality of papaya (Carica papaya) cv. ‘Tainung No. 2’. Smoke-water was characterized analysis. This solution was acidic, substantial amounts of plant nutrients, high level of NH4+ which was important nitrogen sources for plant growth and has a high content of antifungal activity especially phenolic compounds. The compounds of smoke-water were investigated by gas chromatography-mass spectrometry (GC-MS), and 30 compounds were identified, which were mainly alcohols, lactones, aldehydes, acid, ketones, alkaloid and hydroxybenzenes. Among the identified compounds, have 3 types of compounds which considerable in this study, theirs were 2(5H)-Furanone that may stimulate the germination of papaya seeds, phenolic compounds that may inhibited the growth of several fungi and 1H-Imidazole,1-methyl-4-nitroso-5-phenyl- that ability to inhibit and antifungal pathogens. In the germination experiments, low concentrations of smoke-water (0.1% or 0.2%, v/v) not only promoted the maximum rate of germination but also shortened the germination time. Analysis of longitudinal sections of seeds treated with smoke-water concentrations of 0.1% or 0.2% v/v suggested that smoke-water could overcome water impermeability barriers, since it stimulated the seed coat to rupture and allowed the radical to elongate and emerge faster. In the growth experiments, smoke-water promoted multiple growth attributes, such as chlorophyll content and seedling vigor index, at all concentrations in papaya seedling production. Smoke-water was tested in vitro and in vivo for efficacy against damping-off caused by a soil borne fungi and investigate the effect of smoke-water on the morphology of Pythium sp. in scanning electron microscopy observation (SEM). Smoke-water inhibited the mycelia growth and oospore production of Pythium sp. Furthermore, after 3% smoke-water spray application, the morphology of Pythium sp. revealed loss of structural integrity, abnormal degradation, deformation, autolysis, cytoplasmic leakage and further hyphal slimming. In pot experiments, the percentage of papaya plants showing symptoms of damping-off was reduced by all concentration of smoke-water treatments. Fruit rots diseases are the major cause for the postharvest loss of papaya fruit. The control of anthracnose and Phytophthora fruit rots disease, caused by Colletotrichum gloeosporiodes and Phytophthora sp., respectively, by using smoke-water or chitosan tested in the vitro and in vivo. In vitro studies, using water agar amended with smoke-water or chitosan. The results showed that smoke-water or chitosan inhibited fungal growth of C. gloeosporioides or Phytophthora sp. In vivo studies, papaya fruits were inoculated with C. gloeosporioides or Phytophthora sp. and treated with 10% smoke-water, 20% smoke-water, 0.5% chitosan, a combination of 0.5% chitosan and 10% smoke-water, or distilled water (control), The results suggested that the use of 10% smoke-water, 0.5% chitosan or a combination of 0.5% chitosan and 10% smoke-water reduce the lesion diameter and disease incidence of anthracnose and Phytophthora fruit rots disease on papaya fruits. However, 0.5% chitosan coating was more effective among treatments. Postharvest quality, papaya fruits were treated with smoke-water, chitosan, and a combination of chitosan and smoke then were stored at the ambient temperature of 25 ◦C for 9 days. Papaya fruits coated with smoke-water increased the rates of respiration and ethylene production, and caused the papaya to ripen faster than the other treatments. This suggests that smoke-water show promising results for improving seed germination, seedling growth and controlling some plant disease. In addition, smoke-water can possibly economize the use of commercial chemical fertilizers, fungicide, making it a feasible technology for organic farming and non-chemical pesticide management model.

Twenty four Jersey calves were randomly blocked according to sex and birth date, to determine the effects of supplementing Carica papaya seed (Linn) meal (CPSM) on health and growth performance of calves. In the first study, calves were fed CPSM for only 2 days to determine faecal pathogen population. In the second study; calves were randomly assigned at birth to three treatments and fed until weaning at 42 days. Each treatment had six calves. Treatments were; 1) a control group, which did not receive additive; 2) a group that was supplemented 5 g/d of Carica papaya seed meal (CPSM) and 3) the last group that was supplemented 5 g/d of a commercial product containing Lactobacillus acidophilus (Lact). After receiving milk with colostrum from their dams for 3 consecutive days after birth, calves had commercial starter pellet and fresh water ad lib. Treatments were added to whole milk from day 4 and fed to calves before being allowed to suckle from the dams for 30 min three times a day (08:00; 12:00 and 17:00). Calf starter dry matter intake (DMI) was evaluated daily while body weight (BW) and body structural growth were measured weekly. Faecal samples were collected directly from the rectum on day 7 and 10, before and after receiving CPSM respectively. The DMI; DMI/BW; BW and BWG of calves did not differ among treatments, and averaged 305.4 g/d; 7.7 g/d per BW; 39.4 kg and 32.1 kg, respectively. The initial BW, 22.2 ± 1.49 kg did not differ among groups, but the weaning BW was higher (p<0.05) for CPSM fed calves compared to control diet fed calves. It did not differ between control and Lact calves. Calves in Lact and CPSM treatments had similar average daily gain (ADG) and heart girth (HG), which were higher (p<0.05) than calves fed the control treatment. Calves fed CPSM had higher (p<0.05) hip width (HW) and shoulder height (SH) than control calves. There were effects of time (p<0.001) for starter DMI/BW, ADG and all structural body parameters, and effects (p<0.05) of interaction between time and treatments for only starter DMI/BW, ADG and HG. The CPSM treatment reduced faecal coliforms and E. coli (p<0.05) by 93.6% and 96.1%, respectively; and tended to reduce Enterobacteriaceae (p=0.056) by 96.4%. The present study revealed that feeding CPSM to calves during the pre-weaning period increased growth performance by improving average daily gain, feed efficiency and enhancing health status due to low faecal pathogen count
Agriculture, Animal Health and Human Ecology
M. Sc. (Agriculture)