Cryopreservation of plants
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Cryopreservation is the process of freezing and storing biological material at ultra-low temperatures (such as -196⁰C) in liquid nitrogen (LN2) for unlimited periods. Thanks to the application of this biotechnology, all biological... more
Cryopreservation is the process of freezing and storing biological material at ultra-low temperatures
(such as -196⁰C) in liquid nitrogen (LN2) for unlimited periods. Thanks to the application
of this biotechnology, all biological activities including the biochemical reactions
leading to cell death and DNA degradation discontinue. From this point of view, cryopreservation
biotechnology focuses on preservation of cells that have many applications in the
fields of human and veterinary medicine, agriculture and aquaculture. In addition, this form
of biotechnology also has many applications in biomedical research, specifically in the areas
of immunology, virology, neurobiology, toxicology and the pharmaceutical industry.
Cryopreservation Biotechnology in Biomedical and Biological Sciences describes principles and applications
of cryopreservation biotechnology in different research areas and includes seven
chapters that have been written by experts in their research fields. The book chapters are
divided into four sections.
Section I, "Mechanism of Cryopreservation" contains one chapter entitled "Cryoprotectants
and their usage in cryopreservation process", which describes the importance and mechanism
of the cryopreservation process and also its comparison to vitrification. In addition, the
functions and physical and chemical properties of cryoprotectants are discussed.
Section II, "Application of Cryopreservation in Human Medicine Researches" is divided into
four chapters. The first chapter, "Clinical outcomes of assisted reproductive techniques using
cryopreserved gametes and embryos in human medicine", provides valuable information
regarding fertility treatments in human reproductive medicine, focusing on
cryopreservation of spermatozoa, oocytes and embryos. The second chapter, "Cryopreservation
of platelets: advances and current practices" focuses on the long-term preservation of
platelets, which is necessary for the coagulation of blood and stopping bleeding. The third
chapter, "Cryopreservation of preantral follicles", provides valuable information regarding
the structure and development of ovarian follicles. Beside this, advantages and necessities of
using oocytes in preantral follicles for the aim of cryopreservation are explained in humans
and mammals. The fourth chapter, "Vitrification: Fundamental principles and its application
for cryopreservation of human reproductive cells", is related to mechanisms of cryopreservation
in terms of application of this technique in human medicine with emphasis on
reproductive cells. In this way, mechanisms underlying the problem of the intra- and extracellular
ice formation, the role of cryoprotectants, stages of the warming and cooling process
and use of this biotechnology in the field of IVF industry are explained.
Section III, "Application of Cryopreservation in Veterinary Medicine Research", contains one
chapter entitled "Biological signals of sperm membrane resistance to cryoinjury in boars",
discusses biological mechanisms before the cryopreservation/vitrification process causing
cryoinjuries in sperm cells in boars.
Section IV, "Application of Cryopreservation in Agricultural Research", contains one chapter
entitled "Cryopreservation protocols for grapevine shoot tips", which presents valuable information
regarding cryopreservation of shoot tips of grapevines, which is an economically
important agricultural product.
This book covers different applications of cryopreservation biotechnology. I hope that this
book will be helpful for researchers studying cryobiology and related issues. I would like to
thank all the authors for their distinguished contributions, IntechOpen Publishing Company,
and its Author Service Manager Ms. Dolores Kuzelj for her help in publishing this book.
(such as -196⁰C) in liquid nitrogen (LN2) for unlimited periods. Thanks to the application
of this biotechnology, all biological activities including the biochemical reactions
leading to cell death and DNA degradation discontinue. From this point of view, cryopreservation
biotechnology focuses on preservation of cells that have many applications in the
fields of human and veterinary medicine, agriculture and aquaculture. In addition, this form
of biotechnology also has many applications in biomedical research, specifically in the areas
of immunology, virology, neurobiology, toxicology and the pharmaceutical industry.
Cryopreservation Biotechnology in Biomedical and Biological Sciences describes principles and applications
of cryopreservation biotechnology in different research areas and includes seven
chapters that have been written by experts in their research fields. The book chapters are
divided into four sections.
Section I, "Mechanism of Cryopreservation" contains one chapter entitled "Cryoprotectants
and their usage in cryopreservation process", which describes the importance and mechanism
of the cryopreservation process and also its comparison to vitrification. In addition, the
functions and physical and chemical properties of cryoprotectants are discussed.
Section II, "Application of Cryopreservation in Human Medicine Researches" is divided into
four chapters. The first chapter, "Clinical outcomes of assisted reproductive techniques using
cryopreserved gametes and embryos in human medicine", provides valuable information
regarding fertility treatments in human reproductive medicine, focusing on
cryopreservation of spermatozoa, oocytes and embryos. The second chapter, "Cryopreservation
of platelets: advances and current practices" focuses on the long-term preservation of
platelets, which is necessary for the coagulation of blood and stopping bleeding. The third
chapter, "Cryopreservation of preantral follicles", provides valuable information regarding
the structure and development of ovarian follicles. Beside this, advantages and necessities of
using oocytes in preantral follicles for the aim of cryopreservation are explained in humans
and mammals. The fourth chapter, "Vitrification: Fundamental principles and its application
for cryopreservation of human reproductive cells", is related to mechanisms of cryopreservation
in terms of application of this technique in human medicine with emphasis on
reproductive cells. In this way, mechanisms underlying the problem of the intra- and extracellular
ice formation, the role of cryoprotectants, stages of the warming and cooling process
and use of this biotechnology in the field of IVF industry are explained.
Section III, "Application of Cryopreservation in Veterinary Medicine Research", contains one
chapter entitled "Biological signals of sperm membrane resistance to cryoinjury in boars",
discusses biological mechanisms before the cryopreservation/vitrification process causing
cryoinjuries in sperm cells in boars.
Section IV, "Application of Cryopreservation in Agricultural Research", contains one chapter
entitled "Cryopreservation protocols for grapevine shoot tips", which presents valuable information
regarding cryopreservation of shoot tips of grapevines, which is an economically
important agricultural product.
This book covers different applications of cryopreservation biotechnology. I hope that this
book will be helpful for researchers studying cryobiology and related issues. I would like to
thank all the authors for their distinguished contributions, IntechOpen Publishing Company,
and its Author Service Manager Ms. Dolores Kuzelj for her help in publishing this book.
This study investigated optimisation of media and primary-protocorm development stages to enhance secondary-protocorm production as a novel means for propagation of terrestrial orchids, including taxa of conservation concern. Seeds of... more
This study investigated optimisation of media and primary-protocorm development stages to enhance secondary-protocorm production as a novel means for propagation of terrestrial orchids, including taxa of conservation concern. Seeds of Caladenia latifolia were germinated asymbiotically on ½-strength Murashige and Skoog (MS) medium fortified with 5% (v/v) coconut water. Resulting protocorms at 3, 5 and 7 weeks of growth were subcultured to protocorm-proliferation media treatments consisting of ½-strength MS basal-salts medium with 6-benzylaminopurine (BA) and a-naphthaleneacetic acid (NAA) singly or in combination. Conversion of seeds to primary protocorms was high (87–92%). The highest percentage of secondary-protocorm proliferation was 40.1%, using 5-week-old protocorms (early Stage 4 of protocorm development) as explants and cultured on ½-strength MS with a combination of 5 mM NAA + 2 mM BA. Half-strength MS media containing a single plant-growth regulator (BA or NAA) were substantially less effective (<10% protocorm proliferation). The present study has provided a novel approach to sequential protocorm production that will be of value particularly for threatened orchids with limited seed availability. Protocorm proliferation in vitro enables a renewable supply of protocorms with which to conduct propagation, cryostorage and pilot restoration programs.
Despite many reports on regeneration of Vitis after cryopreservation, there is no cryopreserved collection of its germplasm. Some Vitis genotypes are reported to be recalcitrant to cryopreservation. Droplet vitrification, considered to be... more
Despite many reports on regeneration of Vitis after cryopreservation, there is no cryopreserved collection of its germplasm. Some Vitis genotypes are reported to be recalcitrant to cryopreservation. Droplet vitrification, considered to be an emerging generic method of cryopreservation, has been applied only to a limited extent in Vitis. In the present study, we first tested the toxicity of plant vitrification solution in both axillary and apical buds in six diverse Vitis accessions. Droplet vitrification was then applied using 50 % predicted survival time of apical and axillary buds in vitrification solution after pre-treatment of donor plantlets with salicylic acid, a substance known to have a protective role in abiotic stress responses. Results showed that axillary buds are more tolerant of vitrification solution than apical buds and required longer treatment time. Pre-treatment of donor plantlets with 0.1 mM salicylic acid resulted in a significantly higher protection to cryopreserved buds, but serial dehydration in sucrose alone had little effect. Pre-treatment with salicylic acid enabled successful cryopreservation of previously recalcitrant rootstock 41B, albeit at a low regeneration rate. For other genotypes, cryopreservation of 6–11 explants will be sufficient to regenerate at least one plant at 95 % probability. This is the first report of successful cryopreservation of a set of diverse Vitis genotypes by droplet vitrification, and we show that pre-treatment of donor plantlets with salicylic acid is critical for the success. This research will contribute to conservation of Vitis germplasm in a cost effective way avoiding the risks associated with field-based collections.
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