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1

Isolated microspores as a source doubled haploids of barley

100%
Oleszczuk S., Zimny J.
Biotechnologia
|
2001
|
issue 1
139-142
EN
In vitro techniques for doubled haploids (DH) production allow for obtaining homozygous lines in a single generation. This is connected with shorter breeding cycle of the new variety. DH lines have a potential for being used in the selection of recombinants, stabilising of transformed lines and molecular mapping. DH lines are produced from isolated microspores through haploid embryogenesis. Microspore culture has several advantages over anther culture: it reduces the time of cultures, enables monitoring of the earliest phase of embryogenesis, allows for direct development embryos, facilitates the in vitro selection and mutation, allows for avoiding regeneration from somatic anther tissues. Moreover, microspore culture appears to be a promising tool in genetic manipulations (transformation, mutagenesis) and it can be used as a source of protoplasts and suspensions. Here we report on how to induce microspore embryogenesis, resulting in plant formation. The switch of microspore development from gametophytic to sporophytic pathway has been stimulated by various stress factors like cold and heat shock, starvation. Stress treatment not only stops pollen development but also re-programmes the microspore towards embryo formation. The effects of various parameters including pretreatment, carbohydrates and nurse culture have been investigated. After optimising the culture conditions we were able to regenerate high number of fertile plants.
2

Obtaining carrot (Daucus carota L.) plants in isolated microspore cultures

88%
Gorecka K., Kowalska U., Krzyzanowska D., Kiszczak W.
Journal of Applied Genetics
|
2010
|
vol. 51
|
issue 2
141-147
EN
Microspores were cultured on the modified B5 liquid medium containing 2.4D (0.1 mg L?1), NAA (0.1 mg L?1), L-glutamine (500 mg L?1), L-serine (100 mg L?1), and sucrose (100 g L?1). The developmental stages of microspores and divisions were observed. Initially, the formation of binuclear and multicellular structures was noticed. Plants regenerated in the cultures in which the tetrad stage of microsporogenesis had predominated. Embryoids were still forming 24 weeks after the cultures were set up. Six weeks after the transfer of androgenetic embryos onto the B5 regeneration medium, they were converted into complete plants. Out of 90 androgenetic plants planted in a growth chamber, 42 plants adapted to the new conditions. All of those plants proved to be diploids in cytometric analysis.
3

Microspore culture of winter oilseed rape (Brassica napus L.) in conjunction with other in vitro technologies

75%
Cegielska-Taras T., Szala L., Bartkowiak-Broda I.
|
EN
Microspore culture in conjuction with other technologies such as selection, mutagenesis and transformation has been used for the production of novel genotypes of Brassica napus L. for crop improvement. The example of in vitro selection of microspore - derived embryos includes: a) ploidy level, b) seed oil composition (for example: high level of erucic acid), c) genotypes with restorer gene for CMS-ogura system (by means of isozyme marker PGI-2 ), d) herbicide resistant forms. Efficiency of microspore mutagnesis has been tested by the treatment of freshly isolated microspores with UV and MNU. Direct delivery of foreign gene to the microspores (microprojectile bombardment) combined with the use of Agrobacterium tumefaciens to microspore derived embryos seems to be a promising way of oilseed rape transformation.
4

Transformation of microspore-derived embryos of winter oilseed rape (Brassica napus L.) by using Agrobacterium tumefaciens

75%
Cegielska-Taras T., Pniewski T., Szala L.
Journal of Applied Genetics
|
2008
|
vol. 49
|
issue 4
343-347
EN
Haploid microspore-derived embryos (MDEs) constitute a unique material for the introduction of new traits into winter oilseed rape (Brassica napus). MDEs have been transformed by using Agrobacterium tumefaciens strains EHA105 and LBA4404, both carrying the binary vector pKGIB containing the uidA gene encoding -glucuronidase (GUS) and the bar gene as a marker of resistance to phosphinotricin. Transformed embryos expressed GUS and regenerated plants that were resistant to herbicide Basta, as confirmed by a leaf-painting test. Progeny plants of the transformant T-39 were all transgenic, as they inherited T-DNA from their doubled haploid parental plant. Southern-blot analysis confirmed the integration and transmission of T-DNA into T1 plants. Transformation of MDEs facilitates the obtaining of winter oilseed rape homozygous for the introduced genes.
5

Microspore embryogensis ? the stress induced change of gametophytic into sporophytic development

75%
Cegielska-Taras T.
Biotechnologia
|
2004
|
issue 2
12-16
EN
Microspores cultured in vitro can be reprogrammed to divide and produce a bipolar embryo. The reaction to stress treatment is a signal for inducing the sporophytic pathway, preventing the development of fertile pollen grain ? the gametophytic pathway. The ultimate goal is to convert each microspore from a heterozygous F1 plant to a doubled haploid plant so that a population of doubled haploids fully represents the genetic variability of the preceding meiosis.
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