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Doctoral Thesis
DOI
https://doi.org/10.11606/T.11.2019.tde-12082019-120622
Document
Author
Full name
Marina de Lima Nogueira
E-mail
Institute/School/College
Knowledge Area
Date of Defense
Published
Piracicaba, 2019
Supervisor
Committee
Azevedo, Ricardo Antunes de (President)
Borges, Karina Lima Reis
Labate, Carlos Alberto
Souza, Lucas Anjos de
Title in English
Investigating the tomato tolerance to cadmium (Cd) via high throughput proteomics
Keywords in English
Gell free
Heavy metal
Nutrient accumulation
Oxidative stress
Abstract in English
The problem of cadmium (Cd) toxicity aggravates every year. However, the amount of plant species presenting some degree of tolerance to this metal is low. Therefore, it is of utmost importance to understand the Cd tolerance mechanisms. Two tomato cultivars exhibiting distinct levels of tolerance to Cd were investigated by exposing them to 35 μm de CdCl2 for eight days. The samples (roots, stems, and leaves) were collected at 0, 2, 4, 6 and 8 days. The root length and surface area were obtained using a WinRhizo PRO 2009c image analyzer. The ICP OES method was used for the quantification of metals in the different tissues. The translocation index (TrI), metal uptake and Cd distribution nutrient absorbed (NA), and nutrient efficiency (NUE) were calculated. The plant material used for stress indicators and proteomics analysis were collected in liquid nitrogen. Lipid peroxidation and H2O2 contents were obtained by spectrometry. For the proteomics data the gel-free method was used. The stress indicators showed the variation in both cultivars, evidencing the stress caused by the Cd. Chlorosis and necrotic spots were observed in plants exposed to Cd less significant way in the first days of exposure and more evident in the less tolerant cultivar (TTO) in the last days. The growth reduction can be observed in both cultivars, more evident in TTO. Regarding the Cd distribution, the two cultivars showed similar behavior, with the highest percentage of Cd in the roots. The Cd absorption could interfere with both cultivars in the accumulation absorption and efficiency in the use of most nutrients assessed. In general, the Cd interference was more evident at TTO than at YOSH. Highest percentages of TrI were also observed in TTO cultivar. Identification and evaluation of the differentially accumulated protein (DAP) in the two cultivars roots it is possible to observe that in the presence of Cd the cultivar TTO generates a more significant alteration in the amplitude of the accumulation and the number of proteins. These results suggest that the cultivar, which presents a higher degree of tolerance, has some form to make the Cd unavailable or a mechanism that makes the Cd transportation to the leaves more difficult.
Title in Portuguese
Investigando a tolerância de tomate ao (Cd) via proteômica de alto rendimento
Keywords in Portuguese
Acúmulo de nutriente
Estresse oxidativo
Gell free
Metais pesados
Abstract in Portuguese
O problema da toxicidade do cádmio (Cd) agrava-se a cada ano. No entanto, a quantidade de espécies de plantas que apresentam algum grau de tolerância a este metal é baixa. Portanto, é de suma importância entender os mecanismos de tolerância do Cd. Com esse intuito duas cultivares de tomate (TTO e YOSH) exibindo níveis distintos de tolerância ao Cd foram investigadas, expondo-as a 35 μm de CdCl2 por oito dias. As amostras (raízes, caules e folhas) foram coletadas aos 0, 2, 4, 6 e 8 dias. O comprimento da raiz e área de superfície foram obtidos usando um analisador de imagem WinRhizo PRO 2009c. O método ICP OES foi utilizado para a quantificação de metais nos diferentes tecidos. O índice de translocação (Trl), a absorção de metal e a distribuição de Cd dos nutrientes absorvidos (NA) e eficiência de nutrientes (NUE) foram calculados. O material vegetal usado para indicadores de estresse e análise proteômica foi coletado em nitrogênio líquido. A peroxidação lipídica e o teor de H2O2 foram obtidos por espectrometria. Para os dados proteômicos, foi utilizado o método livre de gel. Os indicadores de estresse mostraram a variação nas duas cultivares, evidenciando o estresse causado pelo Cd. A clorose e manchas necróticas foram observadas em plantas expostas ao Cd de maneira menos significativa nos primeiros dias de exposição e mais evidentes no cultivar menos tolerante (TTO) nos últimos dias. A redução do crescimento pode ser observada em ambas as cultivares, mais evidente no TTO. Quanto à distribuição de Cd, as duas cultivares apresentaram comportamento semelhante, com o maior percentual de Cd nas raízes. A absorção de Cd pode interferir nas duas cultivares na absorção, acúmulo e eficiência no uso da maioria dos nutrientes avaliados. Em geral, a interferência do Cd foi mais evidente no TTO do que no YOSH. As maiores porcentagens de TrI também foram observadas na cultivar TTO. Identificação e avaliação da proteína diferencialmente acumulada (DAP) nas raízes das duas cultivares é possível observar que na presença de Cd a cultivar TTO gera uma alteração mais significativa na amplitude do acúmulo e no número de proteínas. Estes resultados sugerem que a cultivar, que apresenta um maior grau de tolerância, tem alguma forma de tornar o Cd indisponível ou um mecanismo que dificulte o transporte de Cd para as folhas.
 
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Publishing Date
2019-08-14
 
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