OCR A2 F215 CLONING IN PLANTS AND ANIMALS Specification: a) Outline the differences between reproductive and non-reproductive cloning b) Describe the production of natural clones in plants using the example of vegetative propagation in elm trees c) Describe the production of artificial clones of plants from tissue culture d) Discuss the advantages and disadvantages of plant cloning in agriculture e) Describe how artificial clones of animals can be produced f) Discuss the advantages and disadvantages of cloning animals DEFINITION OF A CLONE A clone is a group of genetically identical genes, cells or whole organisms Genetically identical means that individuals in the clone have identical DNA with identical chromosome numbers Cloning is a natural process for growth and asexual reproduction Natural cloning of cells involves : DNA replication in cells, mitosis in eukaryotic cells (or binary fission in prokaryotic cells) followed by cell division Cloning can also be achieved artificially To produce identical DNA copies during cloning, it is assumed that no mutation occurs during the process REPRODUCTIVE AND NON-REPRODUCTIVE CLONING Reproductive cloning is the cloning of new organisms with the same genotype of the existing organism Non-reproductive cloning is the production of genetically identical cells, tissues and organs, so that damaged tissues and organs can be replaced. Some examples are the regeneration of cardiac muscle tissue after a heart attack and the repair of the spinal cord in those paralysed by a broken neck/back 1 NATURAL CLONING IN PLANTS Natural cloning in plants is called asexual reproduction or vegetative propagation. This occurs in a number of different ways. Some examples are given below. Examples (1) and (2) are for interest only. Example (3) is on the specification 1. Growing potato plants from potato tubers Potato tubers are the swollen underground stems that we eat as the vegetable. Each tuber is able to produce shoots and roots and will grow into a new potato plant if planted in soil 2 2. Growing strawberry plants from strawberry runners After producing strawberry fruits, the strawberry plant sends out horizontal stems along the ground, called runners. New plants grow at the nodes, along the stem. Initially, the new plant obtains its nutrients from the parent plant. When the new plant has leaves and can produce its own organic nutrients, the runner dies back 3. Vegetative propagation of elm trees by root suckers The English Elm (Ulmus procera) is one of our native trees that reproduces asexually by vegetative propagation English Elms developed from a variety of elm brought to Britain from Rome in the first century A.D. Although English Elms produce flowers and fruits, the fruits contain no seeds and therefore new Elm trees cannot be produced by sexual reproduction Instead, Elm trees reproduce by developing structures known as suckers from their roots. These are shoots that grow from meristem tissue below ground level, usually from roots Each root sucker can grow into a new tree All English Elm trees are clones of each other English Elms are genetically isolated from other Elm varieties because they cannot interbreed with them. English Elms can only reproduce asexually 3 Dutch Elm Disease and Devastation of the English Elm Population in Britain In 1967, a new virulent strain of an Elm disease fungus arrived in Great Britain on imported timber Beetles that lived under the bark of elm trees spread the fungus The saws used to cut down dead branches were not sterilised after use. When the saws were used to prune healthy trees, these trees were infected Approximately 25 million elm trees, most of the English Elm population, died within a few years of the arrival of this fungus The dying Elm trees responded by producing root suckers that grew into new trees Unfortunately, the new trees were clones of the original and being genetically identical to the parent tree, also lacked resistant genes to the disease. There was no variation on which natural selection could act. The new Elm trees did not survive and died from Dutch Elm disease also 4 Why Dutch Elm Disease led to such a Rapid Loss of English Elm Trees All trees were genetically identical Therefore, all trees were susceptible to this disease; none were resistant Beetles are the vector of the disease and can fly from infected tree to healthy tree to spread the disease The beetles only live on the target tree, that is the English Elm Since the trees only reproduce asexually from root suckers, new trees are very close together in clonal patches Tree surgery (cutting out infected branches) was used as a control measure. However, these attempts at control contributed to the spread of the disease. As more trees became infected then more tree surgery was necessary and more saws/surgery equipment were contaminated Dead leaves left on the ground also carry the fungus and are a source of infection for young trees 5 ADVANTAGES OF ASEXUAL REPRODUCTION It is a rapid method of reproduction since only one parent is required Large numbers of offspring can be produced to rapidly colonise an area All offspring are genetically identical to the parent and can therefore thrive in their existing environment DISADVANTAGES OF ASEXUAL REPRODUCTION Offspring have no genetic variation and will have the same less favourable features as the parent If the environmental conditions change (as they did for the elm trees) the cloned organisms may not be able to evolve and adapt to survive ARTIFICIAL CLONING IN PLANTS Farmers and plant growers have been propagating plants artificially for many years. Note that methods (1) and (2) below are for interest only. Method (3) is on the specification. (1)Taking Cuttings Stem, leaf or root cuttings are taken to clone grape vines and other crop plants. This method is used on a large scale in some plant nurseries 6 (2) Grafting A shoot section of a woody plant with desirable features (eg a fruit tree or rosebush) is joined to an already growing root stock. The grafted plant grows into a fruit tree/rosebush that is genetically identical to the parent plant that provided the shoot section with desirable features. The rootstock is different genetically (3) Artificial Propagation in plants using Tissue Culture (on the specification) Definition of Tissue Culture: The production of plants from a small amount of plant tissue that is initially grown in or on a sterile nutrient medium Features of the Use of Tissue Culture This technique can be used to produce very large numbers of genetically identical plants from a small amount of plant tissue from just one, or very few parent plants with desired features Tissue culture is used to produce plants that do not naturally reproduce asexually or are difficult to grow from seeds It is used for the production of more expensive and rare plants such as orchids It can be carried out at any time of year, in any country as it is started in a laboratory. Plants are only transferred into soil in glasshouses when they are well developed 7 Procedure for Tissue Culture 1) Small separate groups of meristematic cells are removed from a plant. These groups of meristematic cells are called explants 2) The explants are dipped into a bleach solution to sterilise the surface of the cells 3) The explants are placed in a sterile aerated nutrient growth medium solution containing: glucose (energy source) amino acids vitamins inorganic ions such as K+, Mg2+ , PO4- NO3plant growth substances such as auxin and cytokinin to stimulate cell growth and division 4) The undifferentiated cells in the explants divide repeatedly by mitosis to produce separate masses of undifferentiated/totipotent cells each called a callus 5) Each callus can be subdivided into smaller groups of callus cells that can be transferred onto sterile agar jelly containing a mixture of plant growth substances that cause the development of shoots and then roots. This technique is called sub-culturing 6) Inducing root or shoot growth involves changing the plant hormone ratio. Root growth requires a high auxin to cytokinin ratio (100 auxin to 1 cytokinin). Shoot growth requires 4 auxin to 1 cytokinin 7) All the procedures described require aseptic conditions to prevent growth of bacteria, fungi and algae in the culture media 8 8) When the plants are large enough, they can be transferred to a less controlled environment and grown in soil in a greenhouse to grow further ADVANTAGES AND DISADVANTAGES OF PLANT CLONING IN AGRICULTURE Advantages A quick method to produce a large number of offspring plants Disease- free stock can be produced All plants have the same features/phenotype eg grow at the same rate/grow to the same height Can be used to reproduce infertile plants Can reproduce plants that are difficult to grow from seed 9 Can produce plants from calluses that have been genetically modified Production is not determined by the seasons. Tissue culture techniques can be carried out in any season and anywhere in the world Small plantlets can be transported easily (including air freighted) and grown in a small space The techniques can be used to culture rare species to save them from extinction Disadvantages Tissue culture is expensive and labour intensive Whole batches can fail if there is microbial contamination There is no or very little genetic variation Therefore, all offspring are susceptible to the same diseases, pests and changing environmental conditions such as none of the plants are drought resistant ARTIFICIAL CLONING IN ANIMALS Reproductive Cloning – the production of whole new animals by cloning techniques The techniques of reproductive cloning are commonly used in the selective breeding of animals, such as dairy cattle and sheep Two Methods of Cloning Animals 1) Splitting Early Embryos (16-32 undifferentiated cells) A female with the desired features is treated with hormones so that she super-ovulates (produces many egg cells at the same time) The eggs are collected and fertilised in vitro (in a petri dish) by sperm, from a desirable male Cells from the embryos produced are separated into several groups. Each group of cells can produce a new genetically identical individual Each embryo can be implanted into a surrogate mother. These surrogate mothers are less valuable cows in terms of desirable features. These cows will have been treated with reproductive hormones to ensure that their uteruses are ready to receive an embryo 10 The embryos develop normally and are born in the usual way. The female with the desirable features, from which the embryos were developed, is not put at risk during pregnancy and is available for more super-ovulation All the calves are clones of each other, but not clones of their surrogate mothers or of either parent that provided the gametes. Why not? 2) Somatic Nuclear Transfer – using enucleated eggs This method was used to produce Dolly the sheep, the first mammal to be produced in this way by cloning A nucleus from a body cell (somatic cell) of one adult mammal, is inserted into an enucleated egg cell (an egg cell with its nucleus previously removed) of another adult female of the same species, using electric shock stimulation. This is called somatic nuclear transfer The diploid cell produced is grown on in vitro to produce an early embryo 11 This early embryo consisting of undifferentiated cells, can be divided into several smaller groups of embryonic cells Each small embryo can be placed into a surrogate mother’s uterus to complete development. In this way, several offspring can be produced using several surrogate mothers. The offspring are clones of each other and of the parent that donated the original somatic cell Label the haploid and diploid nuclei in this diagram. Indicate where electric shock treatment is used. Add diagrams to show how several cloned offspring could be produced from one embryo. Indicate where meiosis and mitosis occur. 12 Table of Advantages and Disadvantages of Cloning Animals by Reproductive Cloning (compared with traditional breeding techniques) ADVANTAGES The number of animals with the required characteristics is increased (if a question indicates details of these characteristics, be specific in your answer and refer to them) DISADVANTAGES Animals are genetically identical – and unable to adapt to changing conditions in their environment. These animals are therefore more susceptible to disease The reproductive rate of a desired animal is increased – many offspring produced in a short time Cloning success rate is very poor The technique can be used to breed endangered animals, without requiring a fertile female of the species. The technique is labour intensive and needs more technology – more expensive The technique can be used to produce transgenic animals (genetically modified ) Cloned animals may have a shorter life span. Dolly was not particularly healthy and died at a relatively young age. The body cell used for the nuclear transfer technique is from an older sheep. Its DNA therefore is chronologically older than the newly born offspring sheep. DNA does degrade as it ages and cells have a limited capacity for cell division (the Hayflick limit) Animals are being denied their natural instincts and behaviour – ethical objections. (note that in a recent examination question of this topic – reference to these ethical objections was not on the mark scheme) Examples include transgenic cows that produce milk with human factor IX and transgenic goats that produce milk with a spider silk protein used to produce bullet – proof vests Avoids the risks of mating males and females (physical damage and transfer of disease) 13 Non- Reproductive Cloning in Animals – the production of genetically identical animal cells and tissues Uses of Non-Reproductive Animal Cloning Laboratory testing of cells Cells from humans and other animals are cultured in vitro to test the effects of new drugs to investigate the involvement of genes or external factors in the development of diseases such as cancer or diabetes In gene therapy Research in this area remains experimental. It involves incorporating a dominant allele into cells of a patient that lack the allele and suffer a disease, as a consequence. The genetically modified cells are cloned in tissue culture to produce a mass of cells that are transferred into the patient’s body. This is covered in the F215 pack on Genomes and Gene Technologies Use of stem cells to treat diseases Stem cells are undifferentiated cells that are able to undergo mitosis to form cells that can become differentiated and specialised. It is hoped that stem cells can be used to treat: Insulin-dependent diabetes, by replacing β cells in the Islets of Langerhans Parkinson’s disease by replacing dopamine-producing cells in the brain. The problem is obtaining stem cells, since there are very few true stem cells in adults (most of their stem cells cannot differentiate into any cell type). A useful source is very young embryos where most or all of the cells are undifferentiated and able to form any cell type in the body. These embryonic stem cells are said to be totipotent (able to form any cell type in the body) or pluripotent (able to form most cell types in the body) 14 Table of Advantages and Disadvantages of Non-Reproductive Cloning ADVANTAGES DISADVANTAGES Tissues/organs will be genetically identical to the individual and will not be rejected Ethical objections to using human embryonic cells in this way. The embryo is a potential human and has rights Avoids the reliance of waiting for an organ transplant No understanding of how cloned tissues/organs will behave over time Cloned cells are totipotent (undifferentiated) and can be used to produce any cell type Religious objections to cloning techniques 15