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Master's Dissertation
DOI
10.11606/D.11.2018.tde-20181127-161058
Document
Author
Full name
Sandremir de Carvalho
Institute/School/College
Knowledge Area
Date of Defense
Published
Piracicaba, 1989
Supervisor
Title in Portuguese
Androgênese em arroz (Oryza sativa L.): indução de calos e regeneração de plantas
Keywords in Portuguese
ANDROGÊNESE
ARROZ DE SEQUEIRO
CULTURA DE ANTERA
Abstract in Portuguese
Com o objetivo de estudar uma metodologia para a cultura de antera de arroz de sequeiro, com variedades brasileiras Dourado Precoce e IAC-165, o presente trabalho foi realizado no Laboratório da Seção Radiogenética do CENA (Centro de Energia Nuclear na Agricultura), USP, Piracicaba, São Paulo. As panículas foram coletadas quando as anteras apresentavam micrósporo no estádio uninucleado e pré-tratadas a 7°C por 7 dias. As anteras foram inoculadas no meio MS modificado e incubadas no escuro a 27°C. Para as duas variedades estudadas, foram analisados os seguintes parâmetros: (1) freqüência de calos produzidos e (2) capacidade de organogênese. Para o estudo da indução de calos, foram pesquisados os seguintes fatores: ferro, sacarose, vitaminas (tiamina, m-inositol e biotina), aminoácidos (glicina e metionina), reguladores de crescimento (2,4-D, NAA e BAP), agar e carvão (fatores químicos); temperatura e tempo de pré-tratamento a frio, temperatura de incubação e irradiação com raios gamas nas panículas e das anteras junto com o meio de cultura (fatores físicos); estádio do micrósporo (fator fisiológico). Para a regeneração de plantas foi utilizado o meio MS e os seguintes fatores foram avaliados: reguladores de crescimento (NAA e cinetina), extrato de levedura, agar, carvão ativado e idade do calo. Os resultados obtidos permitem concluir que: (1) a frequencia da indução de calos aumenta no meio líquido com 56,6 mg de ferro, 4% de sacarose, 4 mg/l de glicina para ambas as variedades e 0,8 mg/l de tiamina para Dourado Precoce e 0,4 mg/l para IAC-165; (2) o meio básico MS modificado foi superior ao meio N6 ; (3) no meio MS modificado, as melhores combinações de 2,4-D e NAA foram 2 mg/l e 2. mg/l para Dourado Precoce e 1 mg/l e 3 mg/l, respectivamente, para IAC-165; (4) pré-tratamento a 10°C por 10 dias das panículas com micrósporo no estádio uninucleado deu o melhor resultado na indução de calos; (5) incubação das anteras no escuro a 30° C aumentou a frequência na indução de calos; e (6) o efeito da irradiação com raios-gamas foi favorável quando anteras da variedade Dourado Precoce foram irradiadas junto com o meio de cultura. Com relação a regeneração de plantas, os melhores resultados foram observados no meio R6 e quando foram inoculados calos com 7 dias de idade
Title in English
Androgenesis in rice (Oryza sativa L.): induction of callus and plants regeneration
Abstract in English
In order to study a methodoloy for anther culture of two Brazilian varieties of upland rice, Dourado Precoce and IAC-165, the present work was carried out in the Laboratory of Radiation Genetics Section of CENA (Centro de Energia Nuclear na Agricultura), USP, Piracicaba, São Paulo, Brazil. The panicles were collected when anther were at uninucleated stage of microspore and pre-treated at 7°C for 7 days. The anthers then were inoculated on the modified MS medium and incubated in the dark at 27°C. For the two varieties studied, the following parameters were analyzed: (1) frequency of callus production and (2) organogenesis capacity. To study the induction of callus, the following factors were considered: iron, sucrose, vitamins, (thiamine, m-inositol and biotine), aminoacids (glycine and methionine), growth regulators (2,4-D, NAA and BAP), agar and activated charcoal (chemical factors); temperature and time of cold pre-treatment, incubation temperature and irradiation with gamma-rays of panicles and anthers together with medium of culture (physical factors); microspore stage (physiological factor). For plant regeneration, the MS medium was used and the following factors were evaluated: growth regulators (NAA and kinetin), yeast extract, agar, activated charcoal and callus age. According to the results obtained, we can conclude that: (1) frequency of callus induction increases on liquid medium with 56,6 mg/l of iron, 4% of sucrose, 4 mg/l of glycine for the both varieties and 0, 8 mg/l of thiamine for Dourado Precoce and 0,4 mg/l for IAC-165; (2) the basic MS medium modified was superior to the N6 medium; (3) as for the MS medium modified, better combinations of 2,4-D and NAA were 2 mg/l and 2 mg/l for Dourado Precoce and 1 mg/l and 3 mg/l, respectively, for IAC-165; (4) pre-treatment at 10°C for 10 days of panicles with microspore of uninucleate stage gave the best results in the induction of callus; (5) anther incubation in the dark at 30°C increased the frequency of callus induction; and (6) irradiation effect with gama-rays was favorable when anthers of the variety Dourado Precoce were irradiated together with the culture medium. Regarding regeneration of plants, the best results observed on the R6 medium and when callus as with 7 days of age inoculated.
 
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2018-11-27
 
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