Citrus somatic hybridization with potential for improved blight and CTV resistance

Mendes, Beatriz Madalena Januzzi; Filho, Francisco de Assis Alves Mourão; Mello-Farias, Paulo; Benedito, Vagner A.

doi:10.1007/s11627-001-0086-y

Cited by 42 publications

(24 citation statements)

References 25 publications

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“…Genetic manipulation of citrus in Brazil has focused on the production of somatic hybrids through protoplast fusion (Glória et al, 2000a(Glória et al, , 2000bMendes et al, 2001).…”

Section: Introductionmentioning

confidence: 99%

Agrobacterium-mediated genetic transformation of 'Hamlin' sweet orange

Mendes

Boscariol

Filho³

et al. 2002

Pesq. agropec. bras.

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The development and optimization of efficient transformation protocols is essential in new citrus breeding programs, not only for rootstock, but also for scion improvement. Transgenic Hamlin sweet orange (Citrus sinensis (L.) Osbeck) plants were obtained by Agrobacterium tumefaciens-mediated transformation of epicotyl segments collected from seedlings germinated in vitro. Factors influencing genetic transformation efficiency were evaluated including seedling incubation conditions, time of inoculation with Agrobacterium and co-culture conditions. Epicotyl segments were adequate explants for transformation, regenerating plants by direct organogenesis. Higher percentage of transformation was obtained with explants collected from seedlings germinated in darkness, transferred to 16 hours photoperiod for 2-3 weeks, and inoculated with Agrobacterium for 15-45 min. The best co-culture condition was the incubation of the explants in darkness, for three days in culture medium supplemented with 100 µM of acetosyringone. Genetic transformation was confirmed by performing ß-glucoronidase (GUS) assays and, subsequently, by PCR amplification for the nptII and GUS genes.Index terms: Citrus sinensis, epicotyls, seedlings, transgenics, breeding methods. Transformação genética de laranja Hamlin via AgrobacteriumResumo O desenvolvimento e otimização de protocolos eficientes de transformação genética é essencial nos programas atuais de melhoramento de citros, tanto para porta-enxertos, como para copas de valor comercial. Plantas transgênicas de laranja Hamlin (Citrus sinensis (L.) Osbeck) foram obtidas pela transformação genética de segmentos de epicótilo, coletados de plântulas germinadas in vitro, com Agrobacterium tumefaciens. Foram avaliados fatores que influenciam a eficiência da transformação genética, como: condições de incubação das plântulas utilizadas para coleta de explantes, tempo de inoculação com Agrobacterium e condições de co-cultivo. A regeneração de plantas a partir de segmentos de epicótilo ocorreu em alta freqüência, por organogênese direta. A maior porcentagem de plantas transgênicas foi obtida utilizando-se explantes coletados de plântulas germinadas no escuro e posteriormente transferidas, por 2-3 semanas, para condições de 16 horas de fotoperíodo, e infectados com Agrobacterium por um período de 15-45 minutos. As melhores condições de co-cultivo foram a incubação dos explantes no escuro, por três dias, em meio de cultura suplementado com 100 µM de acetoseringona. A transformação genética foi confirmada pelo teste histoquímico para ß-glucoronidase (GUS) e, posteriormente, pela amplificação de DNA, por PCR, para detecção dos genes nptII e GUS.Termos para indexação: Citrus sinensis, epicótilo, plântulas, transgênicos, métodos de melhoramento.

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“…Genetic manipulation of citrus in Brazil has focused on the production of somatic hybrids through protoplast fusion (Glória et al, 2000a(Glória et al, , 2000bMendes et al, 2001).…”

Section: Introductionmentioning

confidence: 99%

Agrobacterium-mediated genetic transformation of 'Hamlin' sweet orange

Mendes

Boscariol

Filho³

et al. 2002

Pesq. agropec. bras.

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“…(Calixto et al, 2003) foram obtidos de folhas jovens de plantas cultivadas em casa de vegetação. Procedimentos relacionados ao isolamento, purificação, fusão e cultivo de protoplastos foram realizados de acordo com protocolos anteriores (Mendes et al, 2001;Costa et al, 2003).…”

Section: Methodsunclassified

Regeneração de plantas após fusão de protoplastos de tangelo 'Page' e toranja 'Lau Tau'

et al. 2007

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RESUMO -Buscou-se a hibridação somática entre tangelo 'Page' e toranja 'Lau Tau' visando à produção de porta-enxerto semelhante à laranja-azeda, por esta espécie ser considerada um provável híbrido entre C. reticulata e C. grandis. Após isolamento, fusão e cultivo de protoplastos, obtiveram-se brotações que foram enxertadas in vitro, em laranja 'Hamlin'. Dezessete plantas foram aclimatizadas em casa de vegetação. A análise de citometria de fluxo confirmou a constituição diplóide dessas plantas. Marcadores moleculares RAPD das plantas regeneradas apresentaram padrão de bandas similar ao de tangelo 'Page'. Entretanto, todas as plantas apresentaram conformação fenotípica diferente dos genitores. Termos para indexação: citros, cultura de tecidos, hibridação somática, porta-enxerto, melhoramento genético. PLANT REGENERATION AFTER PROTOPLAST FUSION OF 'PAGE' TANGELO AND 'LAU TAU' PUMMELOABSTRACT -This work aimed to produce somatic hybrid between 'Page' tangelo and 'Lau Tau' pummelo in an attempt to regenerate a similar rootstock to sour orange, because this species is considered a probable hybrid between C. reticulata and C. grandis. After protoplast isolation, fusion and culture regenerated shoots were in vitro grafted on 'Hamlin' sweet orange. Seventeen plants were acclimatized in a greenhouse. Citometric flow analyses revealed that all plants are diploid. RAPD molecular markers of regenerated plants had the same pattern as compared to 'Page' tangelo. However, all plants had phenotypic traits different from both genitors.

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“…In Brazil, citrus protoplast fusion approaches have been developed since 1995, leading to the production of somatic hybrids for rootstock improvement aiming at the control of important diseases, such as blight and citrus tristeza virus (CTV) (Mendes-da-Glória et al, 2000;Mendes et al, 2001;Latado et al, 2002;Costa et al, 2003).…”

Section: Hibridação Somática Entre Citrus Sinensis (L) Osbeck E C Gmentioning

confidence: 99%

Somatic hybridization between Citrus sinensis (L.) Osbeck and C. grandis (L.) Osbeck

Calixto

Filho

Mendes

et al. 2004

Pesq. agropec. bras.

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This work had as objective to produce citrus somatic hybrids between sweet oranges and pummelos. After chemical fusion of sweet orange embryogenic protoplasts with pummelo mesophyll-derived protoplasts, plants were regenerated by somatic embryogenesis and acclimatized in a greenhouse. The hybrids of 'Hamlin' sweet orange + 'Indian Red' pummelo and 'Hamlin' sweet orange + 'Singapura' pummelo were confirmed by leaf morphology, chromosome counting and molecular analysis. These hybrids have potential to be used directly as rootstocks aiming blight, citrus tristeza virus, and Phytophthora-induced disease tolerance, as well as for rootstocks improvement programs.

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Citrus somatic hybridization with potential for improved blight and CTV resistance

Cited by 42 publications

References 25 publications

Agrobacterium-mediated genetic transformation of 'Hamlin' sweet orange

Agrobacterium-mediated genetic transformation of 'Hamlin' sweet orange

Regeneração de plantas após fusão de protoplastos de tangelo 'Page' e toranja 'Lau Tau'

Somatic hybridization between Citrus sinensis (L.) Osbeck and C. grandis (L.) Osbeck

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