Abstract
Somatic embryogenesis is the best demonstration of totipotency in higher plants in which somatic cell produce whole plant like zygotic embryo. It is also demonstrated that immature, weak, hybrid or sometimes inviable embryos can be saved through in vitro culture to prevents its degradation. It may help to cross the reproductive barriers when interspecific hybrids developed. Brasssica is an economically valuable oil yielding and vegetable crop and India is the largest producer of oil seed rape in the world. Various factors affect the embryo rescue in Brassica like growth stage of the embryos, types and composition of the rescue medium etc. The embryo regeneration potential can improve through the modification of culture conditions in both zygotic as well as somatic embryo. Except the embryo culture other parts like ovule, ovary culture can also be done to developed interspecific hybrids. This chapter is focused on the embryo rescue techniques in the genus Brassica and summarizes possible ways of improving the technique used.
Keywords
- Brassica
- embryo rescue
- hybrids
- embryo culture
- somatic embryos
1. Introduction
The embryogenesis in higher plants is controlled by many genes and related embryos specific proteins. Loss of function mutants were used to identified the genes involved in There are about 220
2. Zygotic embryos, genetic embryos and somatic embryos
In higher plants, formation and development of embryo are the two distinct phenomena that can takes place inside the ovule. An ovule has different parts like chalaza, nucellus, micropyle, integuments, and most important embryo sac. The embryo sac develops after the reduction division of the megaspore mother cell. Only one haploid megaspore cell give rise to the birth of embryo sac that’s contains many cells like synergid cells, antipodal cells and one egg cell. Instead of all these cells there is a central nucleus (So called because it is not surrounded by distinct cell wall). The egg cell is polar and contain a distinct nucleus on cytoplasm-rich chalazal pole while there are vacuoles at micropylar end [14]. Egg cell when fertilized called the zygote and gives rise to the embryo. The two important process alternate in each generation, the meiosis (reduction division resulting two haploid cells called the gametophyte) and the fertilization (fusion of the two-haploid nucleus called the sporophyte). The three process are distinct with one another. Formation of zygote after fertilization, formation of the embryo and development of the embryo. Failure of any process disturbed the embryogenesis and cease the formation and development of embryo. After the formation of complete embryo successfully the mature embryo enters in desiccated and metabolically quiescent state [15] or it undergo the period of dormancy that’s complete the process. The dormancy is the process of adaptation to withstand unfavorable conditions. The zygotic embryo in the angiosperm after maturation develops into seeds and it is composed of several tissues, including the embryo, the endosperm, and the testa. The mature embryo has the bipolar axis, on which root and shoot meristem are present and gives rise to the root and shoot during plant development. In contrast to the zygotic embryo, the genetic embryo is formed without fertilization through the process termed as apomixis. Apomixis refers to the formation of embryo in the ovule from the somatic cells. These somatic cells when diploid i.e. nucellus or integument directly gives rise to the embryo and termed as sporophytic apomixis. However, when embryo sac originates either from megaspore mother cells by mitosis or incomplete meiosis in diplospory is termed as gametophytic apomixis.
Totipotency is the most spectacular demonstration of potencies in the cells of higher plants. The somatic embryogenesis is the generation of bipolar structure from any somatic cell that have distinct root and shoot pole. It is very rare phenomenon present in nature and restricted to some plants like
3. Embryo culture
In
4. Embryo rescue in Brassica
Embryo rescue is an
The survival rate of the embryos can be increased when rescued because wide hybridization crosses fail to complete normal sexual reproduction cycle. In
5. Factors affecting embryo rescue
Various factors affect the embryo rescue in
5.1 Nurse tissue
The early stage embryo culture during rescue is very difficult to culture on artificial medium. The nurse tissue provides a natural condition and nutrition so that the chances of embryo abortion reduces. The best nurse tissue for the embryo culture is endosperm and if it is some days old it efficiently increases the chances of survival of the young embryo. The somatic embryos can be used as nurse tissue for culturing the hybrid zygotic embryos as done in
5.2 Culture medium
There are various nutrients formulations used to culture the embryos during rescue. Murashige and Skoog formulation [21] and Gamborg’s B-5 [20] are most frequently used in
5.3 Silver nitrate
The silver nitrate is another very important ethylene antagonistic component that’s play important role in
5.4 Temperature and light
Embryo culture during embryo rescue influenced with temperature and light and their requirements are also varies in different plants [46]. The low temperature regime is best for the embryo rescue but in some cases at high temperature regime (26.4 °C/10.4 °C embryo rescue through ovule culture method gives better results than low temperature regime period (19.4 °C/4.3 °C) (1.75%) in
6. Cytology of cultured embryos
Several biochemical and molecular changes occurs at cellular level as the embryo formed, grow in length and approaches to the maturity. The developmental studies of the somatic embryos showed that the lipid, protein and polysaccharides produced during at varying degrees when embryos were cultured on ABA containing medium. At first one or two weeks polysaccharides were produced and after that polysaccharides lipid and protein accumulated [49]. The analysis of the cultured embryos cells of oil yielding crop like
7. Genes involved in embryogenesis
When a somatic cell enters into embryogenic state it starts to modify the gene expression level. There are number of genes involved in somatic embryogenesis, showed increase expression. These can be categorized in various groups [55]. These are hormone responsive genes, house-keeping genes, genes expressed during maturation, genes coding for extracellular proteins, homeotic genes, HSPs,(Heat-Shock Proteins) germins, zygotic mutants and genes for signal transductions, except these there are the genes of transcription factors. The various transcription factors regulate several genes that showed expression in the somatic embryo induction [15]. In
(AtRaptor1) is responsible for the early development of embryo [62]. The genes also essential for the post-embryonic plant growth because the AtRaptor1A, AtRaptor1B double mutants are defective in meristem driven growth during post embryonic stage [63].
In near future, the techniques of somatic embryos induction, and embryo rescue in
8. Conclusion
In this chapter, the somatic embryos and zygotic embryos with embryo rescue techniques are discussed. The hybrid embryo rescue technique in
Acknowledgments
The author acknowledges Prof. M. Z. Abdin lab, Jamia Hamdard, New Delhi for carried out research work on somatic embryogenesis in
Acronyms and abbreviations
Late Embryogenesis Abundance BABY BOOM gene Leafy cotyledon Abscicsic acid insensetive3 Agamous-like15 WUSCHEL Somatic embryogenesis receptor-like kinase
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