Citrus cybrid regeneration following cell fusion between nucellar cells and mesophyll cells

Saito, Wataru; Ohgawara, Toshifumi; Shimizu, Jun-ichi; Ishii, Shigetaka; Kobayashi, Shozo

doi:10.1016/0168-9452(93)90091-d

Cited by 47 publications

(31 citation statements)

References 15 publications

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“…Protoplastos de citros têm sido obtidos de tecido foliar (Kobayashi et al, 1991;Saito et al, 1993) e de calos e células em suspensão (Oliveira,1993), sendo estes os responsáveis pela regeneração de plantas, utilizando-se de soluções enzimáticas constituídas de celulase e pectoliase (Carneiro et al, 1998), dissolvidas em metade das concentrações dos macronutrientes do meio MT (Murashige & Tucker, 1969), sais minerais de CPW (Frearson et al, 1973) ou em ácido 2-[N-morfolino]-etanossulfônico (MES) e NaH 2 PO 4 (Grosser & Gmitter Junior, 1990b).…”

Section: Introductionunclassified

Isolamento e eficiência de plaqueamento de protoplastos de citros

2002

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As recentes ferramentas biotecnológicas para o melhoramento in vitro de citros incluem a hibridação somática por fusão de protoplastos. A otimização dos protocolos de isolamento, plaqueamento e cultura de protoplastos, fundamental para maior eficiência na produção de híbridos somáticos, foi avaliada em 11 variedades cítricas. As soluções enzimáticas testadas foram: 1. celulase Onozuka RS, 1%; macerase R-10, 1%, pectoliase Y-23, 0,2%; 2. celulase Onozuka R-10, 0,2 %; macerase R-10, 0,3%; driselase, 0,1%; 3. celulase Onozuka R-10, 1%; macerase R-10, 0,2%; driselase, 0,1%. O plaqueamento dos protoplastos foi realizado em meio de cultura EME 0,7 M, nas densidades de 2 x 10(4); 5 x 10(4); 10(5); 2 x 10(5) e 3 x 10(5) protoplastos.mL-1, no escuro, a 25 ± 1°C. A solução enzimática 1 possibilitou melhor rendimento no isolamento de protoplastos para a maioria das variedades, exceto para o limão-'Cravo', onde o melhor rendimento foi obtido na solução enzimática 3, e para as laranjas-doces 'Valência' e 'Succari', que apresentaram maiores rendimentos na solução enzimática 2. A eficiência final de plaqueamento, avaliada aos 90 dias de cultivo, foi superior nas densidades de 10(5) e 2 x 10(5) protoplastos.mL-1, para todas as variedades. O desenvolvimento de embriões somáticos foi observado em todas as variedades, exceto para tangor 'Murcote'.

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Section: Introductionunclassified

Isolamento e eficiência de plaqueamento de protoplastos de citros

2002

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“…We investigated the nuclear and cytoplasmic DNA of the regenerated plants and showed that all of the somatic hybrids possessed mitochondrial genomes of nucellar cells [4,10] . Furthermore we found regeneration of cybrids which possessed nuclear genomes of mesophyll cells and mitochondrial genomes of nucellar cells [11] . Here we describe establishment and application of Citrus somatic hybridization system, and utilization of cybrids for somatic hybridization [12].…”

Section: Introductionmentioning

confidence: 79%

“…In the fusion experiment between Sudachi (C. sudachi) and lime (C. aurantifolia) , 4 out of 12 regenerated clones were cybrid. In another experiment of sudachi and lemon 4 out of 6 clones were cybrids [11] . Thereafter, high frequencies of cybrid regeneration in the similar somatic hybridization methods were reported [23][24][25] .…”

Section: Application Of the Somatic Hybridization System To Citrus Brmentioning

confidence: 94%

Production of Somatic Hybrids and Cybrids in the Rutaceae Family and Application to Citrus Breeding.

Ohgawara

Saito

Kobayashi

1997

Plant Biotechnology

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“…Subsequent analyses based on molecular markers of these plants have indicated that they are actually alloplasmic hybrids or cybrids. Most of these plants arose from interspecific combinations (Deng et al 1993Saito et al 1993Saito et al , 1994Yamamoto and Kobayashi 1995;Moriguchi et al 1997). Microcitrus confers resistance to both biotic stresses, including burrowing nematode and Phytophythora, and abiotic stresses such as drought and flood.…”

Section: Introductionmentioning

confidence: 98%

“…Many unexpected diploid plants have been regenerated following standard symmetric fusion between diploid species (Saito et al 1993(Saito et al , 1994Grosser et al 1996;Liu et al 1999. These plants were called leaf-parent-type ones because they were morphologically identical to the leaf parents used in the fusion.…”

Section: Introductionmentioning

confidence: 99%

Molecular characterization of the nuclear and cytoplasmic genomes of intergeneric diploid plants from cell fusion between Microcitrus papuana and Rough lemon

et al. 2002

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Simple-sequence-repeat (SSR) and PCR-RFLP were employed to characterize the nuclear and cytoplasmic genomes of intergeneric diploid plants derived from symmetric fusion between Microcitrus papuana Swingle and Rough lemon (Citrus jambhiri Lush). Three out of five SSR primers distinguished the fusion parents from each other and the regenerated plants showed band profiles completely identical to the leaf parent, Rough lemon. Amplified products from the intergenic regions of cpDNA between trnD-trnT were digested with HaeIII and MspI, and those between trnH-trnK were digested with HinfI, and both the regenerated plants and Rough lemon shared the same band patterns, which were different from the embryogenic parent, M. papuana. With mtDNA, only 2 out of 12 primer pair/restriction enzyme combinations (nad4 ex 1-2/TaqI and nad4 ex 1-2/HindIII) revealed polymorphisms between the fusion parents. With the former combination the regenerated plants showed the same fragment distribution as that of the embryogenic parent, M. papuana, whereas with the latter, a novel band absent in the fusion parents was detected in all of the regenerated plants, suggesting a possible rearrangement. The present research indicates that the plants analyzed were putative cybrids containing nuclear DNA and cpDNA from Rough lemon and mtDNA from M. papuana. Presumed mechanisms leading to the regeneration of diploid hybrid plants following symmetric fusion are discussed herein.

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Citrus cybrid regeneration following cell fusion between nucellar cells and mesophyll cells

Cited by 47 publications

References 15 publications

Isolamento e eficiência de plaqueamento de protoplastos de citros

Isolamento e eficiência de plaqueamento de protoplastos de citros

Production of Somatic Hybrids and Cybrids in the Rutaceae Family and Application to Citrus Breeding.

Molecular characterization of the nuclear and cytoplasmic genomes of intergeneric diploid plants from cell fusion between Microcitrus papuana and Rough lemon

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