Obtaining autotetraploids in vitro at a high frequency in Citrus sinensis

Zhang, June; Zhang, Min; Deng, Xiuxin

doi:10.1007/s11240-007-9240-5

Cited by 37 publications

(26 citation statements)

References 9 publications

(8 reference statements)

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“…Cells to be treated with colchicine were harvested 7 days after the third suspension subculture. It has been found that a majority of citrus cells are in S-phase of mitotic cycle on days 6 and 7 from the beginning of their subculture (Zhang et al 2007;our observations). This phase corresponds to the stage when DNA synthesis and replication occurs and is ideal for application of colchicine since this antimitotic agent can only act on actively dividing cells (Zhang et al 2007).…”

Section: Resultsmentioning

confidence: 96%

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In vitro production of autotetraploid Ponkan mandarin (Citrus reticulata Blanco) using cell suspension cultures

et al. 2009

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An efficient protocol for colchicine mediated production of in vitro autotetetraploids from Ponkan mandarin using cell suspension cultures is described. Cells were treated with 1 g l -1 colchicine for 4 or 8 days before transfer into solid EME medium supplemented with 5% maltose. Colchicine treated cells were placed in medium with or without an overlay of 1:2 medium-mixture of liquid 0.6 M BH3 medium and 0.15 M EME ? maltose liquid media. It was observed that modifying the immediate cell environment by addition of the liquid overlay played a positive role in cell differentiation and subsequent plant regeneration. Ploidy levels were determined with a flow cytometer and confirmed by chromosome staining using the enzymatic maceration method. A large number of non-chimeric autotetraploids were generated using this method. Such plants have great value in a breeding program for the development of seedless triploid citrus, as very few available tetraploid breeding parents are easy to peel.

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Section: Resultsmentioning

confidence: 96%

“…However, the frequency was low. Recently, Zhang et al (2007) described a protocol to obtain autotetraploid citrus plants using callus and generated a number of sweet orange plants.…”

Section: Introductionmentioning

confidence: 99%

In vitro production of autotetraploid Ponkan mandarin (Citrus reticulata Blanco) using cell suspension cultures

et al. 2009

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“…Since the 1930s, it has been used to increase plant ploidy levels (Blakeslee and Avery 1937). An advantage of colchicine is that it does not reduce its polyploidizing capacities after being autoclaved (Zhang et al 2007). However, in many plant species, colchicine causes side effects such as sterility, abnormal growth, chromosome losses or rearrangements and gene mutation (Luckett 1989).…”

Section: Antimitotic Agents Used In In Vitro Chromosome Doublingmentioning

confidence: 99%

“…It contributes to the effect and toxicity of the treatment. In many studies, antimitotic agents are dissolved in DMSO (Allum et al 2007;Awoleye et al 1994;Greplova et al 2009;Hamill et al 1992;Kermani et al 2003;Khosravi et al 2008;Stanys et al 2006;Subrahmanyam and Kasha 1975;Takamura et al 2002;Väinölä 2000;Yang et al 2006;Zhang et al 2007Zhang et al , 2010a. DMSO increases cell permeability and allows increased absorption of chemicals (Hamill et al 1992).…”

Section: Concentration and Exposure Timementioning

confidence: 99%

Mitotic chromosome doubling of plant tissues in vitro

Dhooghe¹,

Laere²,

Eeckhaut³

et al. 2010

Plant Cell Tiss Organ Cult

351

258

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In vitro chromosome doubling can be induced by several antimitotic agents. The most commonly used are colchicine, oryzalin and trifluralin. The process of induced chromosome doubling in vitro consists of a typical succession of sub-processes, including an induction phase and a confirmation protocol to measure the rate of success. The induction step depends on a large number of variables: media, antimitotic agents, explant types, exposure times and concentrations. Flow cytometry is the pre-eminent method for evaluation of the induced polyploidization. However, alternative confirmation methods, such as chromosome counts and morphological observations, are also used. Since polyploidization has many consequences for plant growth and development, chromosome doubling has been intensively studied over the years and has found its way to several applications in plant breeding. This review gives an overview of the common methods of chromosome doubling in vitro, the history of the technique, and progress made over the years. The applications of chromosome doubling in a broader context are also discussed.

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“…Marie & Brown (1993) propuseram coefi cientes de variação de 1 a 2%, para resultados de Paralelamente aos estudos realizados para a defi nição de um método seguro para a propagação in vitro de genótipos elite de híbridos entre capim-elefante e milheto, a Embrapa Gado de Leite tem trabalhado para a obtenção de materiais hexaploides por meio da duplicação cromossômica de híbridos triploides (Abreu et al, 2006;Barbosa et al, 2007). Como as estratégias adotadas para duplicação cromossômica geralmente utilizam protocolos de exposição de explantes in vitro a agentes antimitóticos (Gu et al, 2005;Yang et al, 2006;Zhang et al, 2007), a obtenção de protocolos de embriogênese somática, para híbridos triploides de capim-elefante e milheto, além de representar uma possibilidade de multiplicação de genótipos elite, representa uma alternativa para indução in vitro de duplicação cromossômica, que visa à obtenção de híbridos hexaploides férteis.…”

Section: Resultsunclassified

Embriogênese somática em híbridos de Pennisetum sp. e avaliação de estabilidade genômica por citometria

Campos

Calderano

Pereira

et al. 2009

Pesq. agropec. bras.

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Resumo -Os objetivos deste trabalho foram estabelecer um protocolo efi ciente de embriogênese somática, em híbridos triploides entre capim-elefante (Pennisetum purpureum Schumach.) e milheto (P. glaucum (L.) R. Br.), e avaliar por citometria de fl uxo a estabilidade genômica das plantas obtidas in vitro. A embriogênese somática e a regeneração das plantas foram estabelecidas a partir de embriões zigóticos maduros de híbridos entre capim-elefante e milheto. Foram testados quatro tratamentos com 2,4-ácido diclorofenoxiacético (2,4-D), nas concentrações 0, 1, 2 e 3 mg L -1 , para indução de calos embriogênicos, e dois tratamentos com inositol a 1 e 2 g L -1 , para regeneração das plantas. Os tratamentos foram dispostos em delineamento inteiramente ao acaso. A combinação ótima de hormônios foi de 2 mg L -1 de 2,4-D, para indução de calos embriogênicos, e de 1 g L -1 de inositol, para conversão de embriões e regeneração de plantas. A análise de quantidade de DNA, por citometria de fl uxo das plantas regeneradas, indicou a não ocorrência de alterações em ploidia durante a embriogênese somática e a regeneração das plantas. A quantidade de DNA nuclear e a ploidia das plantas regeneradas foram estáveis e homogêneas em comparação às das plantas controle. Não ocorreu instabilidade cariotípica no sistema de regeneração usado para híbridos de Pennisetum.Termos para indexação: Pennisetum glaucum, Pennisetum purpureum, cultura de tecidos. Somatic embryogenesis in hybrids of Pennisetum sp. and genomic stability evaluation by cytometryAbstract -The objectives of this study were to establish an effi cient protocol for somatic embryogenesis in triploid hybrids between napiergrass (Pennisetum purpureum Schumach.) and pearl millet (P. glaucum (L.) R. Br.), and to assess the genomic stability by fl ow cytometry of the plants obtained in vitro. Somatic embryogenesis and plant regeneration were successfully established from mature zygotic embryos of napiergrass and pearl millet hybrids. Four treatments with 2,4-dichlorophenoxyacetic acid (2,4-D) at 0, 1, 2 e 3 mg L -1 were tested for embryogenic calli induction and two treatments with inositol at 1 e 2 g L -1 were tested for plant regeneration. The treatments were arranged in a completely randomized design. The optimum hormone combinations were 2 mg L -1 of 2,4-D for embryogenic callus induction, and 1 g L -1 of inositol for embryos conversion and plant regeneration. The analysis of DNA content by fl ow cytometry of the regenerated plantlets indicated that no ploidy changes had been induced during somatic embryogenesis and plant regeneration. The nuclear DNA content and ploidy levels of the regenerated plants were stable and homogeneous in comparison to those of the control plants. There was no occurrence of karyological instability in the regeneration system utilized for Pennisetum hybrid.

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Obtaining autotetraploids in vitro at a high frequency in Citrus sinensis

Cited by 37 publications

References 9 publications

In vitro production of autotetraploid Ponkan mandarin (Citrus reticulata Blanco) using cell suspension cultures

In vitro production of autotetraploid Ponkan mandarin (Citrus reticulata Blanco) using cell suspension cultures

Mitotic chromosome doubling of plant tissues in vitro

Embriogênese somática em híbridos de Pennisetum sp. e avaliação de estabilidade genômica por citometria

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