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Authors
Abstract(s)
A grande maioria das plantas existentes não tolera a presença de elevadas concentrações de
sal no seu meio de crescimento, independente das condições edafoclimáticas, sendo essas
designadas “plantas galicófitas”. Por outro lado, existem plantas que conseguem realizar todo
o seu ciclo de crescimento e reprodução em ambientes com concentrações salinas elevadas,
sendo as mesmas designadas por “plantas halófitas”. Ao longo da costa portuguesa, as plantas
halófitas são frequentemente encontradas em locais com boa drenagem de água, como por
exemplo, salinas, dunas e margens de bacias marítimas.
A Salicornia spp. é uma planta halófita também conhecida por espargo do mar, mede entre 25
a 35 cm e apresenta uma morfologia alongada. Esta planta tem vindo a ser descrita como
benéfica para a saúde, uma vez que contem vitaminas A e C, assim como, cálcio, ferro, iodo,
magnésio, ácido p-cumárico, ácido ferúlico e sódio. À Salicornia estão associados/descritos
efeitos antioxidantes e anti-inflamatórios, sendo que esta planta também está relacionada com
a prevenção de complicações cardiovasculares, como por exemplo a hipertensão.
Tendo em conta os benefícios do consumo da Salicornia spp., tem sido crescente o interesse
comercial e científico no estudo dos compostos e na multiplicação desta planta. Neste sentido,
e apesar de já se encontrarem descritos alguns resultados relativos à multiplicação in vitro da
Salicornia spp., no desenvolvimento desta tese de mestrado, otimizaram-se os processos de
multiplicação pelo método convencional e, pela primeira vez, foram realizados ensaios em
Biorreatores de Imersão Temporária. Os resultados oBITdos culminaram numa metodologia
simples e eficiente de multiplicação in vitro, assim como de pré- aclimatização ex vitro da
Salicornia spp. De seguida identificou-se e quantificou-se o ácido ferúlico, comparando o
composto produzido pelas plantas oriundas da multiplicação in vitro com as plantas
provenientes da fase de pré-aclimatização ex vitro. Para tal, foram realizados diferentes ensaios
em diferentes fases do processo de multiplicação in vitro de plantas: Desinfeção e
Estabelecimento, sendo o tratamento mais eficiente o ET4 (HgCl2 a 10% durante 3 minutos);
Micropropagação pela técnica convencional, analisando-se diferentes parâmetros nos quais
se destaca o tratamento MCT17 (DMS com 8.87 µM BAP + 5.37 µM NAA + 513 µM NaCl) e a
Micropropagação pela técnica de biorreatores de imersão temporária, alcançando-se o
melhor desempenho no tratamento com MBT2 (DMS, 5 minutos de Imersão a cada 6 horas e 1
minuto de ventilação a cada 6 horas); Pré- aclimatização ex vitro, todos os ensaios oBITveram
resultados positivos, não existindo diferenças significativas entre os mesmos.
Por último, quantificou-se o ácido trans-ferúlico existente em amostras provenientes da
multiplicação in vitro e das amostras pré-aclimatizadas obtendo-se respetivamente 3,39±1,26
µg/mL e 4,17±3,48 µg/mL de ácido trans-ferúlico.
The large majority of the existent plants does not tolerate large amounts of salt in their growth medium, regardless the soil and climate conditions, being this kind of plants called of “galiphocite plants”. However, there are plants that can perform all their growth and reproduction cycle in environments with high salt levels (being called halophytic plants). It is important to refer that across the Portuguese coast is very common to find this kind of plants near places with good drainage of water (like e.g. saline, dunes, banks of sea basins). The Salicornia spp. is a halophytic also known as asparagus from the sea, measuring between 25 to 35 cm and presenting an elongated morphology. Moreover, it has been described as beneficial to health since present vitamins A and C as well as calcium, iron, iodine, magnesium, p-cumaric acid, trans ferulic acid and sodium. To Salicornia are linked antioxidants and antiinflammatory effects also, this plant is related with the prevention of cardiovascular diseases. Regarding the benefits of the Salicornia spp. consumption there are commercial and scientific interest in study the compounds and the multiplication process of this plant. Although now there are already described results from the in vitro multiplication of Salicornia spp., there are yet the need to optimize and complete this information. Concerning this, in this work there was optimized the in vitro multiplication conventional method. Moreover, for the first time, these plants were grown using Temporary Immersion Bioreactors which enabled in the production of a full methodology of in vitro multiplication and ex vitro pre-acclimatization of the Salicornia spp. After, the trans ferulic acid was identified, quantified and the amount of this acid produced by plants from in vitro multiplication and by plants from nature was compared. To accomplish the above mentioned, there were performed several experiments in different phases of the in vitro multiplication process: desinfection and establishment, being the most efficient the treatment ET4 regimen (10% HgCl2 for 3 minutes); Micropropagation using a conventional methodology, when different parameters a analyzed and compared the most relevant treatment was MCT17. Micropropagation using the temporary immersion bioreactors technique, the best performance was assigned to MBT2 treatment (DMS, 5 minutes every 6 hours in duration and 1 minute every 6 hours of ventilation); Pre-acclimatization ex vitro, all treatments were successful and there was no difference between treatments. Finally, the trans-ferulic acid existing in the several in vitro and pre-acclimatized samples were quantified, respectively 3.39 ± 1.26 µg/mL and 4.17 ± 3.48 µg/mL of trans-ferulic acid.
The large majority of the existent plants does not tolerate large amounts of salt in their growth medium, regardless the soil and climate conditions, being this kind of plants called of “galiphocite plants”. However, there are plants that can perform all their growth and reproduction cycle in environments with high salt levels (being called halophytic plants). It is important to refer that across the Portuguese coast is very common to find this kind of plants near places with good drainage of water (like e.g. saline, dunes, banks of sea basins). The Salicornia spp. is a halophytic also known as asparagus from the sea, measuring between 25 to 35 cm and presenting an elongated morphology. Moreover, it has been described as beneficial to health since present vitamins A and C as well as calcium, iron, iodine, magnesium, p-cumaric acid, trans ferulic acid and sodium. To Salicornia are linked antioxidants and antiinflammatory effects also, this plant is related with the prevention of cardiovascular diseases. Regarding the benefits of the Salicornia spp. consumption there are commercial and scientific interest in study the compounds and the multiplication process of this plant. Although now there are already described results from the in vitro multiplication of Salicornia spp., there are yet the need to optimize and complete this information. Concerning this, in this work there was optimized the in vitro multiplication conventional method. Moreover, for the first time, these plants were grown using Temporary Immersion Bioreactors which enabled in the production of a full methodology of in vitro multiplication and ex vitro pre-acclimatization of the Salicornia spp. After, the trans ferulic acid was identified, quantified and the amount of this acid produced by plants from in vitro multiplication and by plants from nature was compared. To accomplish the above mentioned, there were performed several experiments in different phases of the in vitro multiplication process: desinfection and establishment, being the most efficient the treatment ET4 regimen (10% HgCl2 for 3 minutes); Micropropagation using a conventional methodology, when different parameters a analyzed and compared the most relevant treatment was MCT17. Micropropagation using the temporary immersion bioreactors technique, the best performance was assigned to MBT2 treatment (DMS, 5 minutes every 6 hours in duration and 1 minute every 6 hours of ventilation); Pre-acclimatization ex vitro, all treatments were successful and there was no difference between treatments. Finally, the trans-ferulic acid existing in the several in vitro and pre-acclimatized samples were quantified, respectively 3.39 ± 1.26 µg/mL and 4.17 ± 3.48 µg/mL of trans-ferulic acid.
Description
Keywords
Ácido Trans-Ferúlico Biorreatores de Imersão Temporária Multiplicação In Vitro Salicornia Spp.