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Master's Dissertation
DOI
10.11606/D.11.2018.tde-20181127-161725
Document
Author
Full name
Adriano Reis Lucheta
E-mail
Institute/School/College
Knowledge Area
Date of Defense
Published
Piracicaba, 2004
Supervisor
Title in Portuguese
Clonagem e análise do gene codificador da enzima acetolactato sintase (ALS) em Amaranthus quitensis e Bidens pilosa
Keywords in Portuguese
AMINOÁCIDOS
CARURU
CLONAGEM
HERBICIDAS
INIBIDORES DE ENZIMAS
PICÃO PRETO
PLANTAS DANINHAS
RESISTÊNCIA GENÉTICA VEGETAL
Abstract in Portuguese
A enzima Acetolactato sintase é a enzima chave na biossíntese dos aminoácidos de cadeia ramificada valina, leucina e isoleucina. A ALS foi identificada como alvo de ação de cinco classes distintas de herbicidas: sulfoniluréias, imidazolinonas, triazolopirimidinas, pirimidil-tiobenzoatos e sulfonilamino-carbonil-triazolinonas. Estes herbicidas vêm sendo intensivamente utilizados no controle de plantas daninhas em diversos países, levando ao surgimento de biótipos resistentes. Até hoje foram observadas 83 espécies resistentes aos herbicidas inibidores da ALS no mundo. Na América do Sul, foram identificadas as espécies Bidens pilosa (picão-preto) e Amaranthus quitensis (caruru), no Brasil e na Argentina, respectivamente. Sementes destas espécies foram coletadas em lavouras de soja para ensaios biológicos e determinação da curva dose-resposta, confirmando a resistência e sensibilidade aos herbicidas inibidores da ALS. A seqüência codificadora do gene da ALS de cinco plantas, dos biótipos resistentes e sensíveis, de ambas as espécies, foi amplificada por PCR, clonada em plasmídeos e seqüenciada. Os resultados mostram que a seqüência codificadora do gene da ALS em A. quitensis é composta por 2010 nucleotídeos tanto nos biótipos resistentes como nos sensíveis. Em B. pilosa, a seqüência dos biótipos resistentes é composta por 1935 nucleotídeos e dos biótipos sensíveis é composta por 1959 nucleotídeos. Foi demonstrado que o fenótipo de resistência em ambas as espécies é determinado pela substituição do aminoácido Triptofano (W), nos biótipos sensíveis, pelo aminoácido Leucina (L), nos biótipos resistentes, na região denominada domínio B. Resultados semelhantes foram descritos em outras espécies. Plasmídeos binários contendo a seqüência codificadora completa da ALS, de biótipos resistentes e sensíveis de A. quitensis e B. pilosa, foram construídos para a transformação de discos foliares de tabaco via Agrobacterium tumefasciens
Title in English
Cloning and analysis of the gene that codifies for the acetolactate synthase enzyme of Amaranthus quitensis and Bidens pilosa
Abstract in English
Acetolactate synthase (ALS) is the key enzyme in the biosynthesis of branched chain amino acids: valine, leucine and isolucine. ALS has been identified as the site of action to five distinct classes of herbicides: sulfonylureas, imidazolinones, triazolopyrimidines, pyrimidinyl thiobenzoates and sulfonylamino-carbonyl-triazolinones. These herbicides have been used intensively to control weed plants in many countries, increasing the appearance of herbicide resistant biotypes. Until today, it has been observed eighty three ALS inhibitors herbicides resistant species in the world. In South America, resistant biotypes of Bidens pilosa (hairy beggarticks) and Amaranthus quitensis(pigweed) were identified in Brazil and Argentina, respectively. Seeds of these species were harvested from soybean fields for bioassays and dose-response curve to confirm resistance and sensitivity to herbicides inhibitors of ALS. The coding region sequence of the ALS gene of five plants from resistant and sensitive biotypes of both species were amplified by PCR, cloned into plasmids and sequenced. The data shows that the A. quitensis coding region of the ALS gene is composed by 2010 nucleotides of both resistant and sensitive biotypes. In B. pilosa, the resistant biotype is composed by 1935 nucleotides and the sensitive one is composed by 1959 nucleotides. It was demonstrated that the resistance phenotype in both species is due to a Tryptophane (W) aminoacid exchange, in the sensitive ecotype, to a Leucine (L) in the resistant one, at the domain B region. Similar results has been described for others resistant species. Binary plasmids were constructed containing the complete codified ALS sequence of A. quitensis and B. pilosa of resistant and sensitive biotypes to transform tobacco leaves by Agrobacterium tumefasciens
 
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Publishing Date
2018-11-27
 
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