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1

In vitro culture of cereal microspores

100%
Oleszczuk S., Zimny J.
Biotechnologia
|
2000
|
issue 2
142-160
EN
The development of microspore culture methods in the Poaceae family has received considerable attention in recent years. Isolated microspore culture can be induced in vitro to switch their development from gametophytic to a sporophytic patway, resulting in embryoid or callus formation. Different stresses like cold or heat shock and nitrogen starvation have been identified as the major trigger in inducing microspore embryogenesis. Microspore culture appears to be a promising toll for future production of double haploids in cereals. Isolated microspore culture has several advantages over anther culture in genetic manipulation and haploid study, such as: direct observation of microspore development, unique possibility to study plant embryogenesis, easier in vitro selection and mutation, easier transformation of single cells. Moreover, isolated microspores are the most efficient way of double haploid regeneration. Many factors such as genotypes, physiological status of donor plants, stage of microspores development, pretreatment of anthers or spikes, method of microspores isolation, culture media, nurse culture and culture conditions, have a great influence on microspore culture. These and other problems concerning in vitro culture of isolated microspore are discussed in this review.
2

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.
3

Consequences of transformation of embryogenic Triticale cell

71%
Zimny J., Czaplicki A., Menke-Milczarek I., Oleszczuk S., Sowa S.
Biotechnologia
|
2000
|
issue 2
64-72
EN
Plants carrying foreign genes have been obtained for many crops including wheat, rice, maize, barley and Triticale. The most important aspect for practical breeding is the regeneration of whole plants from a specific cell possessing the desired agronomic properties. Particle bombardment provided the necessary breakthrough for the efficient transformation of cereals. Efficient regeneration is a prerequisite for all transformation techniques. The aim of the presented work was to study the progeny of transgenic plants of the allohexaploid cereal species Triticale. By combining an efficient regeneration system with the successful particle bombardment method we were able to obtain transgenic Triticale plants. Transgene expression was sometimes unstable and generally resulted in the decline of the expression, although some lines showing stable expression were also selected. In our laboratory several generations of androgenic doublehaploid transgenic lines have been regenerated and multiplicated. The integrated transgenes were detected in Triticale lines by in situ hybridisation method. The stability of trangenes has been studied on ten generations. A regeneration system from single cell to plant combined with microprojectile bombardment appeared to be the most efficient transformation method for Triticale. Numerous chimeric genes are now available for research. Some of these genes may appear useful in the future breeding of Triticale.
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