Plant tissue culture

welcome back friends welcome to be talking about plant tissue culture and actually we have different videos regarding plant tissue culture recorded earlier but I don’t have a kind of compiled version of all those videos to talk about in general what is flank tissue culture because I have been talking about tissue culture a lot and we have separate video series for animal tissue culture and a separate series for plant tissue culture but in this video I am going to give you the basics of plant tissue culture what it means and how we do plant tissue culture and why it’s important so let us begin with it now there are certain things I want to talk about regarding it the first thing I want to talk about it what is what is plant tissue culture now the idea of plant tissue culture arise from a very simple thought that is the plant cells that we see the healthy plant cells without any kind of infection and stuff if you take those plant cells they are capable of dividing and producing a whole plant from wherever you take those plant cells from whatever region you take those plant cells from it could be from the root it could be from the chute region it could be from the bird a lateral but whatever region you take those cells they have the capability of grow and dividing and producing a mature plant sibling and that’s what very very important and in this case that thing we exploit that important property of the plant cells which is also known as totipotency totipotency means total potential of generating as a whole body of organism from a single cell of that organism that’s a beautiful property now our stem cells carry that property that’s why if we culture stem cells that can give rise to different lineage of cells you know from stem cells the lineage of liver cells the lineage of nerve cells and those different varieties of cells will be produced from stem cell varieties now in this case plant cells it’s nothing a special sense like stem cells there are all the cells that are present in the plants have the property of doing it now in our case in animal cell culture we know there are lineage of cells like myeloid progenitor cells lymphoid progenitor cells among the blood so myeloid progenitor cells will give rise to different types of cells lymphoid progenitor cell will give rise to different set of cells so they are known as pluripotent cells that means they are capable of developing multiple different types of cells but here a single cell let us say a growing plant is there I know I don’t have green color today in my hand so you talking about plants is a growing plant now this is this is the tip of that plant so if we take a cell from the tip of the plant and if you plate it in the Culture Media The Culture Media contains all the nutrients that the plant cell required to grow and divide and if you supply those things all those things together and also I played that and pasted that in the environment which supports the growth of the plant then what will we see that that cell will give rise to a mature plant like the previous one that is the idea of plant tissue culture okay so you take out a cell or tissue chunk of cells say from the plant different regions of the plant it could be from the top of this root and will be top of the chute it could be the bottom of the root it could be the leaves it could be the lateral buds whatever region you take you take the cells out place it into the medium in a proper environment they will grow now the second question about the plant tissue culture they always get is what are the conditions that you need to maintain conditions that you need to maintain and how you do that okay now those things are kind of merged because the conditions we maintain is a part of how we do the whole process now in the conditions we have two different things we have the nutrient source and the environment okay we have the source of nutrients and I can say three things source of nutrients then we need the environmental factors and then we also need the growth regulators these are the three things that we require because you know nutrients are obviously required for for any organism to survive so here we need different nutrients forces among the nutrients we have two different types of source macronutrients and micronutrients macronutrients are the nutrients that our plant require more in higher concentration for survive while in micronutrients they require very less of those nutrients to survive the most important nutrient sources are nitrogen source phosphorus source potassium source those are the sources

we call them NPK very very important except for that magnesium is also very important calcium is also very important while the micronutrients part there will be molybdenum and some small other other metallic materials and materials which are not that important but secondary help helping in the secondary permit of light production and other purposes copper and stuff now if you look at here the environmental factors the environment factor consists of the temperature the pH and the light these are the three very very important environmental factors for a plant to grow now the temperature that we always look for depends in all these things the pH pH of the soil or in this case is not a soil but it is medium the Culture Media the pH of the media the osmolarity of that media plays a vital role and also the temperature where we put those plates in where you put those plans to grow and also that the light source because a tree wants light to grow because without light they cannot produce food they cannot grow so in all these cases we need all the stuff right and these things are not generally fixed it’s fixed for a single species of plants for different types of plants those things varied okay and they varied in a different way and in a large scale for different plans to plants in some plant can grow at the temperature of 25 degree Celsius maybe some other plant will love to grow at 29 degree Celsius sometimes we try to have a more neutral pH some may have a slightly acidic pH they love ascetic base to grow so it varies for different types of plants that you choose for the tissue culture process while the well the mostly they have a range between the temperatures that natural temperature where the plant naturally inhabited okay so if you take the habitat of the plant the natural habitat of those plant where you are taking you can see the temperature of that place you can see the the kind of climate there is so you try to mimic those situations in the lab so that that plant can grow better okay the other thing is about the lighting the light source the light is very very important now remember the light is required for their photosynthesis so plant derived the light from Sun right because Sun is a huge source of light there so in this case we can place them in a room where we can add sunlight but it’s not always possible because the labs that we design not always are open access to the sunlight so in those cases what we need to do we need to apply for the idea of artificial light and actually plants can grow very well in those artificial light they don’t require exactly the sunlight to grow instead they require the photons for the photosynthesis so if you can deliver the photons to any artificial light they can also grow so this is the thing but again among this light thing plants have different durations of life you know we call them long day plants some of them we call short day plants and stuff some plants get flowering if the day time is larger than the night time some players get flowering if the night time is longer than the day time so it depends on the plant again you are dealing with what kind of time shift you are going to provide now one thing you should remember by looking at this idea is that the lab where you say the environment that you said it will be fixed for only one type of plant species let’s say you are you want to do the tissue culture of two different species we have the temperature difference of two degree Celsius and one is long day one is short day you cannot deal both of them in the same lab because you have a specific environmental fixed in that lab you know but we use artificial lamp system attached to a circuit who knows after every let’s say it’s they require eight hours of night and rest of them daytime or eight hours of data store them on light so that that artificial light is designed and attached to the circuit in such a way that they will turn on the light for only that eight hours and then turn it off so these are the things that you need to look for scenarios if you look if you go so this is a kind of conditions that we need to monitor the conditions we monitor is in the lab itself while the nutrient sources that we require that will be a part of the medium itself okay so I told you about the curve this this conditions that we require three things environment nutrient and growth regulators that is the third thing growth regulators let’s talk a little bit about the growth regulators growth regulators are nothing but plant hormones because for the plant to grow and divide and as you are doing this thing in the lab you want the plant to grow and divide very rapidly so so you want to add some extra growth regulators from outside don’t need to go on for the plants to develop actually

you are taking a cell or a chunk of cell to develop into a plan so you never know what amount of hormones they are going to produce because they are not going to produce anything you need to keep them supplying all these things so you also need to supply this plant hormones okay this plant hormones the examples of this hormones oxen Zipporah lean cytokine in ethylene abscisic acid these are the five major types of growth regulators that are found in in plant to produce naturally auxiliary lean cytokine in ethylene and abscisic acid they all have different functionalities now I am not going to talk about their functionality and structure in details in this video I have separate video on plant hormones where I talk about all the stuff you can watch that video and that link is provided in the description probably or you can search it in my channel to watch it now you just need to know simple things that aux-in gibberellins and cytokinins mainly first of all oxen and zebra lean plays a vital role or the plant to grow especially oxen it’s known as a growth hormone but it helps sell to divide and grow zebra lien also plays a vital role for the plan to be taller while cytokinin helps this Saito kinase and cell division process cell differentiation process while abscisic acid and ethylene they are kind of end point hormones where they only work for the plant to develop when the plant grows and it matures then abscisic acid actually in larger plants it helps the plants to to remove all its all its you know the leaves and stuff during the autumn and this time and ethylene also helps the fruit to be ripened right so whatever food you you know like I say if you take up let’s say five unripe banana and one ripe banana if you put it there you see all the bananas just get right by overnight so how is that because he theory is a gaseous hormone which can be released from those fruits which are ripening and it can cause other fruits to be ripened very quickly so these are the hormones that we require mainly we require oxen spirulina and cytokine in at the very earlier stage of the plant tissue culture for the cell to grow and divide very rapidly and once they start dividing rapidly they will give us mass of cells in a moment they will give a mass of cells at the very beginning so from one so it will keep on dividing it will give a chunk of cells or mass of cells and that mass of cells are known as callus okay it’s known as callus so once the callus is formed that is the mass of cells of that same type is formed now that callus can give rise to a complete plant and and large not large actually small small part version of that same plant but it’s a fully functional plant from a mass of cells so that thing we can do this is how it actually produce in some cases we do not require to develop into callus but in most of the cases they go through the stage of called callus undifferentiated mass of plant cell just like a ball of cells like tumor so these are the ways to do that so how could you do that you need to have a proper environment set up in the lab you need to have Patriots actually go for Patriots in the plant tissue culture because the plant will grow tall soup a triplet is not enough to hold them so usually go for the tube we call the plant tissue culture tubes they are the fat thicker version of the test tubes like that okay and we also add the cotton plugging things and stuff all this stuff okay there and this is the medium the medium you can use it can be liquid it can be solidified just like the medium that we use in bacterial culture you can add the solidifying edge into the same medium to make it solid so that the plant can grow but remember you the play the medium is not that solid because the plant cells the mass of cells that will grow here slowly start to originate its root it shoot and all the stuff and small tiny plant will come up like that when the plant will come up then you take that open open that up take out this plant and easily can put it into the soil but remember one thing after this whole thing is done I mean the conditions and stuff after you achieve the plant you make the plant you take the plant you cannot directly put it into the natural environment because this plant is not being introduced to the natural soil till now right so it is a better idea to make it habituated with the environment first take the plant and what we do we put it into the artificial environment though we put it in the soil but the soil is properly maintained with all the minerals and water and nutrients

everything then once the plant grows say one to two inch it’s not to ensure three three four inch tall then we take that plant out from there and then we put it into the natural environment the natural soil because the natural soil will not contain this amount of balanced nutrients that we used to provide them during the experiments or during the lab development so this is how the whole process of plant tissue culture can be achieved ultimately the idea is how what are the different types of plant tissue culture that we can now achieve it does not matter where exactly you’re taking the cells you can take the cells where the cells are dividing very very fast for example the shoot tip of region which is the meristematic tissue you know apical meristem attic tissue or you can take the root meristematic tissue from another region and the tip of the root or tip of the chute as these are the meristematic tissues they are growing and dividing very very fast there is another important thing that those meristematic tissues as they’re growing and dividing very fast they don’t catch any sort of infection most of the time so it’s very very good that you take those cells and you allow them to grow but also you can take the cells from this one this bud region as well as the plant Leafs also you can cut the leaves and you can take the cells and do that thing with the leaves also so you can do all this different regions but again it depends on the type of plants you are using now you know once the different set of tissue cultures we do we need to standardize those protocols say you you are using as completely different plant species for tissue culture you don’t know nobody have ever done ever try to do tissue culture with that plant now you need to come up with the idea like you need to check whether if you take the cell from the top of the root it’s going on top of the chute it’s growing or or from the leaf it’s growing it’s up to you which is the most convenient way to grow that plant now once we find that data you need to publish that data and now it will be standardized and anyone else in the field if trying to do the same tissue culture with the same plant they are going to follow your rule there so this is the idea of how we go for tissue culture and stuff okay so based on from where we are taking the cells we can divide them into as a shoot tip culture root tip culture or leaf culture and so on okay but most of them are carry is the same procedure you need to put them there and they will grow another thing I didn’t talk about is obviously the sterilization stages because you are dealing with cells and cells can get infected right so sterilization is obviously an integral part of all the different stages that we’re dealing with now say sterilization of the plates and sterilization of the tubes and stuff we need to do and also we need to see realization of the scalpel and stuff whatever thing you but again one thing is important that plants are much more immune than the animal cells animal cells are much more fragile they don’t they have more prone to be attacked by many different bacteria and protists wasps and all the other cells so the conscience of contamination for animal tissue culture is more compared with the plants because plants are much more resistant against all those infection compared with the animal cells okay so that in a sense is the idea of plant tissue culture now the question is why we do plant tissue culture the final thing I want to talk about is why we go for plant tissue culture now the reason behind plant tissue culture is a simple term I can tell you a single word micropropagation or clonal propagation whatever it is micropropagation and clonal propagation means you take one plant and you want to produce the same plant many number of the same type of plant the easiest way to do with that if you know all the all the machineries and you know another thing plant tissue culture labs are not that costly to set compared with the animal tissue culture which is far more costly and expensive so you can set it easily you need some space and stuff and these are the stuff you can accommodate but if you know how to grow them and if you have a formula to grow them the nutrient sources and all the stuff you can easily take the plant and you can easily grow them and and simultaneously in a lab you can place thousands and thousands of small tiny vessels where you can get all those smaller plants so just imagine you have one plants in the ground you take the cell now you are culturing it thousands and thousands of the copies of the same plant now after the process is done you will implant them in the in the soil so what you will get you get thousands and thousands of the same copy of the plans right that’s the micro propagation or clonal propagation why clonal because we are getting the same clone of plants again and again there so it’s very very fast it’s very rapid to a not not only see not also that but but for understanding

of their nutrient sources their growth properties and how they behave and stuff this is also very important also let us say you you are dealing with a plant which is very very endangered there are very few species left and found one of them you want to preserve that plan teaming to make more copies of that plant this is a very good way to do that because you cannot do the natural breeding there it is not the breeding purpose it is not the breeding here what we are doing your cloning that same cell now once you clone the cells you can do the cell breeding natural breeding or any other breeding types between those plants in the future but this is how you can do the stuff with plant tissue culture it’s interesting and it is really really very important for many different species of plants that we are dealing with okay and let’s say a plant you want to produce a specific product in huge amount but it will take time the plant to naturally grow you can go in this way it will go very very fast so that’s all about plant tissue culture the overview of the basics and if you want to know all the stage in details like should tip culture root tip culture there are my videos about those and the links are provided in the description you can watch them anytime you want so that’s for it guys if you like this video please hit the like button subscribe to my channel to get more and more useful videos like that thank you