CUBE Chaatshala:- The Causerie will still be on!

22nd June 2020 Summaries

Summary of CUBE Chatshaala discussion by @Lydia (22nd June 2020):
WhatsApp Image 2020-06-23 at 3.07.59 PM
(PC: @drishtantmkawale )
(Outline of the summary: credits: @KiranyadavR)

We started with summarizing our discussion that we had in the previous webinar.
Identification of Drosophilidae family characteristics was discussed along with the goof ups. We are now looking for labelled photos of D.melanogaster fruit fly wing from authentic references.

Then optical zooming and digital zooming was then introduced by @⁨Aashutosh Sir⁩.
How to get a resolved photo of a wing of fruit fly at home.
White background with good lightning (from another phone or torch) .

How microscope work??
Can we simulate it??
How image is formed in a Microscope??
@⁨Yash CUBE⁩ & @⁨Arunan Sir⁩ together had built up a conversation on this for all of us.
Yash will give a summary about the principle or the working of a microscope.
We can use transparent (transparent file cover) sheet to put object(fruit fly wing) on transparent sheet and we can use another source of light underneath the transparent sheet to get contrasting effect as discussed.

Taste organs and smelling organs:
Smelling organ in fruit fly: Antenna
Taste organ: Proboscis
Are the chemicals different for taste and smell?

2 other organs that we discussed about:
Arista plumose??
Maxillary palp??

Then we discussed about a situation where there is a good smell with a bad taste.
Banana kept with quinine mixed.
It gives out a very good smell by which the flies are attracted to it…but when it tastes the banana…it’s a bitter taste that it gets… suddenly it moves away from it and repels.
What will happen the next time when this fly gets the smell of banana?
Some of us said that the fly will have a memory of the last time and the fly will repel it even after getting a good smell of banana.

How does this happen?
How is the memory formed?
When the first time the fly tastes the banana with quinine it repels as a reflex action…a sudden action…but simultaneously that message is being passed on to the brain which is stored there as a memory…
Where is this message stored?
How are these messages stored?
And when the next time the fly is being exposed to the same sweet smell… how is it repelling back?
What is being inhibited that was earlier helping the fly to move towards the banana?

This was then taken up by @⁨Manasi CUBE.
Manasi explained the neuronal connection that is happening.
The diagram here was drawn by Manasi during the webinar…it was explained very well…
A good taste sensory neuron is connected to the neuron in the brain.
Manasi can summarize and give some points of that discussion.
Later it was connected to the learning and memory behaviour of snails.
Eric kandel’s work.
This was explained by Anjani CUBE and Manasi CUBE

By Rechel Tirkey from CUBE Ranchi
On 22/06/2020 , In CUBE webinar , discussion on drosophila classification and identifying features of drosophila was going on with the help of pictures shared by @Aswathy CUBE Thriprayar, Nattika then the discussion shifted into the topic microscope , taking pictures, magnification , how to take pictures with good contrast and light ? What is the role of light while taking pictures ?
We discussed principle of microscope and how mirror helps to reflect the light towards object, how objective lens and eyepiece magnify the image of the object .
When we take pictures from our phone camara , we face many problems like as @Drishtant said , most of the time when we try to focus by nearing the camera towards object it get blurry . @Aashutosh Sir @Yash CUBE told about digital zooming and optical zooming , so for good picture we need optical zooming instead of digital zooming.
We also continued the last day’s discussion how flies attract towards smell and taste . @Manasi CUBE Snail and @Anjani CUBE Snail said about how snail attract towards taste and smell , The connection between good smell and bad taste . By giving them good smell and taste to the snail and by observing in behaviour we are trying to understand about neuroplasticity and neurotransmitters .
I wanted to ask that in snail learning taste and smell and learning sense though tentacles is same or not , is this habituation is same as learning smell and taste of food ? I was getting network problem and often getting disconnected .

By Yash Sheregare from CUBE Mumbai:
Discussion on Microscopy Imaging
This idea may not be accurate but please raise questions so that it gets more clear.
Yesterday during discussions we tried to discuss how an image is formed of a specimen using a compound microscope? :microscope:
For our discussion we considered the speciment to be the wing of a fruit fly.
In a microscope our object (specimen) the wing is kept on a transparent glass side
How is an image of the wing formed that we see all the features of the wing using a microscope?
How are images formed using microscope?

  1. 2 breaks in costal vein
  2. incomplete subcostal vein
  3. small anal cell
    Above :arrow_up: Are the features we want to see in the wing.
    We tried to explain that light source in case of the microscope is the mirror placed at the bottom side of the microscope stage.
    The light falls from the bottom side that is reflected from the mirror and the light falls on the object and passes through transparent places and is bent at opaque places.
    This light :arrow_up: travels further from the specimen to the objective lens where the objective lens is adjusted in such a way by moving it upside or down.
    This magnifies and focuses the image and further light travels through the eye piece and falls on to our eyes :eyes:
    And hence we are able to see a magnified image with a good contrast and focus.
    Hence the importance is of the source of light that comes from bottom and travels the lens where there Adjustment of the lens to get a clear focused and magnified image!

By @saswathy679 from CUBE SN College, Nattika
:point_right:How to Get a Clear image of Fruit Flies Wing in our Phone Camera !?

Aashutosh sir is suggests How to get a good focusing image in our Phone camera.He suggested a very Nice idea. First he introduced about the Optical zooming and Digital zooming.what is the difference between Optical Zoom and Digital Zoom.According to Aashutosh Sir the optical zoom is achieved by using your camera’s lens. The digital zoom, on the other hand, is achieved by cropping and enlarging the image once it has been captured by the digital camera’s sensor.we will have more control over what part of the image is cropped and enlarged, and this is the preferred method of achieving digital zoom of our images. Digital zoom often results in lower quality pictures when it is enabled on your camera and we should disable it whenever possible. Most digital cameras allow you to do this easily.
In some reference says On a digital camera, optical zoom is a true zoom feature.This will enable you to get a closer view of the subject before taking your picture.Optical zoom allows for better photo quality than digital zoom.

Take picture in Optical zoom, We get a Very Good Result in Our Phone Camera

By Drishtant M Kawale from CUBE Mumbai:
I will like to add some points to this:
After summarizing briefly on the problems faced by the fruitfly group on the identification of the fruitflies, we came on discussing that how will we be able to make the best use of our mobile camera?

The things which need to be kept in mind while taking a photograph of the wing includes good lighting which can be provided with another mobile phone’s torchlight, learning to use our phone camera is also an important thing.

Few days back in the Webinar, Rahul CUBE TIFR had added that if we want to take our :selfie: selfie, we try to snap it in the best way possible (by taking it multiple times) so that we get satisfied, similarly, here too we should try to take the best picture possible.

Then, Aashutosh Sir (not intended to😜) explained to us the difference between optical zoom and digital zoom of which our phone camera possesses the digital zoom.

Digital zoom means the zoom which we can perform through our phone camera. In this, the lens of our camera doesn’t come closer to the object or specimen, else, it gets adjusted internally.

Optical zoom is something which we see in professional camera where the camera lens can be moved closer and farther to the specimen or object.
Optical zooming is the one which provides us better clarity of the picture where the pixels picture do not stretch much.
Now, as we have a digital zoom facility, we can simulate the optical zoom by bringing the phone camera closer to the specimen physically!
And then try to zoom and then take a photograph.
Then we discussed the principle of microscopy and formation of the image and how can we adjust our surrounding in our to simulate the effect like microscope.

Later, we connected the neurology of the fruitfly with the one of Snails.
As olfactory studies are done in both and share the same basic principle too!

While discussing the memory formation in Snails, Manasi added that memory formation accounts to the formation of new axon terminals beside the existing ones so that more signal can be conveyed.
I didn’t understand it much, we shall look forward to discuss the same today (maybe I misunderstood it).
Manasi also made a statement that habituation (what habituation is it?) is a result of exhaustion of neurotransmitters!
Is it?
The question arising in my mind is, will the neurotransmitter be ever exhausted?
Won’t the cell re-synthesise the neurotransmitter?
For example: Acetylcholine, a lipid based neurotransmitter is synthesised inside the neuron and is then released.
Won’t this process continue to happen?


CUBE Chatshaala Summary on 23rd June 2020 by Drishtant M Kawale from CUBE Mumbai:

Fruitfly Identification
Continuing from yesterday’s discussion on the identification and problems faced while identifying the fruitfly wing, @Aswathy CUBE Thriprayar, Nattika presented a photograph of the wing of a fruit fly from a mixed culture.
We concluded that the resolution of the picture was not clear. By resolution, I mean that the details aren’t getting revealed.
Wing of a fruitfly having characteristics like the incomplete sub-costal vein, two breaks in the costal-vein and presence of anal cell will make sure that the fly is of the Drosophilidae family.
But in @Aswathy CUBE Thriprayar, Nattika’s case, the two breaks are visible but we are not able to locate the anal cell as well as the incomplete sub-costal vein.
Here is where the photography skills will pitch in!

We then discussed the arrangement of the mobile camera so as to get a clear picture of what we want.
@Dibya Sankar, a Physics Research Scholar from TIFR, Colaba suggested that we should try to put a drop of water on our mobile camera lens after which (to my understanding), it shall act as a macro lens which will show us a magnified image.
Dr. Subhojit Sen asked to be careful while putting the drop of water, as it might enter our phone through the camera lens.

As we have been discussing this from a month, it’s time that we shall focus on looking what actually we want. What do we need from this wing?
Drawing a quick conclusion shall help.

Developmental Biology of Fruit fly
As the main point of attraction here is olfaction, we came on the olfaction of the larva.
The questions that raised were:
How does a larva smell?
Is it not peculiar that a crawling creature; larva, undergoes such a modification that within a span of days, it gives rise to an adult fruit fly which instead of crawling, flies!
The wings of the fly emerge from the Imaginal discs which are there in the larvae.

Why are they called Imaginal dics?
Do we have an answer for this?
How does the wing develop from these imaginal discs?

Neurological Relation between Fruit flies and Snails
After @Manasi CUBE Snail and @Anjani CUBE Snail joined us, we discussed the learning behaviour of Snails.
How does habituation take place?
Talking about learning behaviour, we shed some light on the Assay conducted in which Coriander is fed to the Snails and a bitter-tasting compound is adding.

Why is this done?

What is the association between good smell and good taste?
Although smelling and tasting are two different phenomena, does it lead to habituation?
What is the association between good smell and bad taste?
Does the snail form a memory of the bad taste of the compound?

Later, big terms like sensitisation and resensitisation were introduced.
Need to understand them too!

Referring to these, Komal Singh from Bhind, MP added about the types of neurotransmitters - excitatotory and inhibitory which are involved in these processes or in the formation of memory and which decide whether the neuron will pass on the message or not.

Nobel-Prize Winner Eric Kandel’s work on the seaslug Aplysia cannot be left behind if we are discussing the neurology in Snails.
Following this, with the help of a graphic photo, we saw and discussed, what are electrical and chemical synapse?

In the end, @Rechel Tirkey told about the status Moina culture in her home lab and her collaborators’ lab too, their further plans with Moina to approach their research question as well as objective.

I have just highlighted the points, we shall have a discussion on them!


Summary-CUBE Chaatshala 2nd July 2020

Yesterday’s discussion involved both the branches of Biology.

We started with Botany and then discussed Zoology till the end and in between them, we found out a common topic which relates both of them!

The sleep-wake cycle or circadian rhythm studies!
Aashutosh Mule Sir was eager to know the sleep-wake cycle studies were done in the Phyllanthus plant by P. Chitralekha at her place. She has designed a special set-up for studying the same!
The objective is to study the effect of continuous light on the sleep-wake cycle of the plant given that the plant will be exposed to continuous light (white) throughout the day.

How will that help?
We know that as it is a sleep-wake cycle, it is going to repeat itself at some interval of time. So here, the cycle is repeating every 24 hours.
And we come to know that the plant is awake or sleeping by its leaves.

If the leaves of the Phyllanthus plant are closed or turned inside, it is sleeping. And if the leaves are wide open, the plant is awake.
And this opening and closing takes place each day at a particular time in the morning and at evening respectively which may/may not (I don’t have a clear idea of this) be dependent on the sunlight.

Interesting right?

We can believe that us humans or even fruit flies for an instance have a Brain which must be regulating the sleep along with the hormones.

But what about plants?

Do they have a nervous system too?
If not, then how is the sleep regulated?
This is what comes to my mind when I think about the sleep-wake cycle of the plant.

The experiment comprises of control plant to be kept in natural light and the test plant to be kept in the artificial light provided through bulb.

P. Chitralekha had done this experiment last year too and had recorded observations (which she is going to tell us😄) and is planning to do the same this year too around July 19th.
The thing which grabbed her interest is the observations which were made by her. Those were unexpected observations!
So, to confirm it again, she is going to do this.

As P. Chitralekha is known to this plant and has worked on it earlier too, she has some experience. But what will be interesting is that CUBist should also start such kind of experiment and contribute as well as share their data. Then only we will understand how it happens. Otherwise, the knowledge of this will remain confined.

One such CUBist from Goa is Nidhi, who cleared her Class XII examination this year and has been actively participating in the plant science discussions. Nidhi too is working on Phyllanthus plant since one year (and more I guess, don’t know) and has done a similar kind of experiment as mentioned above with the aim to check the effect of different wavelengths of light on the Circadian Rhythm of Phyllanthus plant.

The main objective of both the experiments is to check the effect of light exposure on the sleep-wake cycle, but the only variable here is, P. Chitralekha is using just white led light as the artificial light source whereas Nidhi is using four different coloured lights which are Red, Yellow, Green and Blue. The bulb here used is of 100Watts (the power rating) and covered with cellophane sheet.

Nidhi had kept them under light from 5.30pm to 8.30pm and the control was (same as earlier) kept in natural light.

And then the time of opening and closure of the leaves was noted down.

Please add to this by shedding some more light (:stuck_out_tongue_winking_eye:) on this too and correct me.

As I mentioned earlier, we connected both the Biological Sciences -Botany and Zoology through one common topic in them; the Sleep-Wake Cycle.

A few days back, taking some inspiration from the plant studies, @Saida had hinted that she too is interested in studying the same in Fruit flies!

But before that, another unusual thing happened. Saida , Yash Shergare, Aswathy Suresh, Kshipra have been attempted to study and gather data on the sleep-wake cycle of fruit flies. This is being done by keeping tomato slices as a bait/attractant for the fruit flies and the bait is being observed at the interval two hours and the number of flies on the bait is noted.

In the day time, the peak number of fruit flies is around 35 flies and in the night time, there are comparatively very less number of flies.
Now from this, we can gather a conclusion (by simply relating what happens with us humans) that the flies are active during the day (active implies they are showing movement around the bait, they are mating, courtship is taking place, eggs are layed, etc) and inactive during the night (we humans too, do the same). By inactive I mean to say that there are not much flies, even if they are there, they are stationary on the bait, do not move or mate, etc.

But Saida once found out that in 2am/4am (I don’t remember) in the morning, there were around 5 flies on the tomato bait out of which 3 flies were active! (now we know what do we mean by active)

This is interesting because this is unusual! Instead of sleeping, they are active and moving on the bait whereas the rest 2 flies were inactive. Going deep (referring to the 2017 Nobel-Prize work in Physiology or Medicine), in the fruit fly (and maybe in humans too), at night the accumulation of the per-tim (period and timeless) protein complex induces sleep in the fly and in the morning, when there is sunlight, the per-tim complex is broken down and the fly wakes up from the sleep. This is what happens normally.

But here we saw that the fly (instead of being inactive, sleeping) was moving around the bait!
Why that?
Can these flies be termed as mutants?
Which gene must have been mutated? Period (per), Timeless (Tim), Clock (Clk), Cycle (cyc), Cryptochrome (cyc)?

Can we find out that are these flies actually mutants or is it a chance event?
How can we do that?
@Lydia @Arunan

What shall be the first step by which we can study this?
How would’ve scientists like Seymour Benzer and Ronald Konopka studied the flies? The so-called mutant flies I must say?

Kanishka suggested that to find out whether the flies are mutants or no, the scientists must have done some sophisticated procedures!! (I am not going to name them)

But is so much of sophistication really required?

How can we identify that the fly is mutant or not in a simple, logical way? A way which may or may not be true but at least makes sense!

Aswathy Suresh and Saida are going to start with this.
Please let us know how are you starting!
Put the Plan-of-work and Design!

Later, @Rechel_tirkey added the names of fruit flies found in Jharkhand-Santhali Tribe call it Rhon and the Oraon Tribe calls it Tingli and the place where the Oraon tribes are found is Urukh. Rhon and Tingli is the general name for any fly in these areas.
This was the conclusion.

Please comment and add in your point of view on yesterday’s discussion!


This is a very important question. Elsewhere @Manpreetheersskp, @Arunan and I were discussing, the role of brain and nervous system. For some sea squirts, which do not need to move, they eat up their brain. Now that plants do not need to move, it makes sense to me that they do not require a specialised organ called brain!

@drishtantmkawale, Why do you think plants should have a brain?

But, do plants stop sensing!?
@Manpreetheersskp do sea squirts stop sensing? :thinking::thinking:



And all these activities with Phyllanthus, Tingli/Rhon (thanks @Rechel_tirkey) that you are talking about involving genes and the nervous system appear to have very interesting links! Both seem to play a role in response to changes in the environment (nature)!

Is it just me who feels the same?



YES,plants can sense but they have only few senses :exploding_head:.You should visit the following link for more information @karnamdpdurga

And in case of sea squirt . The sea squirt eats, or to more exact reabsorbs, notochord, tail, sense organs and nervous system, since these are no longer needed, while it feeds by wafting water into its mouth cavity, and filtering out suspended particles.


I think this is the same mechanism that many microbes do. Our CUBist friends can vouch for this. All bacteria, amoeba, paramecia, etc ingest in this particular way! But they respond to the environmental changes, which means they have mechanisms to sense. And none of them have brain as an organ!

So, to all CUBists as well here, what then do you think is the role of nervous system and the brain? Can you please share links to the studies that you’re doing with earthworms and snails for reference here, please?



Screenshot (65)
The whiteboard from CUBE Chatshaala-21 July 2020
We had a good, long discussion on the COVID-19 which has wreaked havoc as a pandemic.

Some of the questions which we tried to answer, build up a logical proposition on;

  • Started with how does the COVID19 enter our lungs?
    This was answered later but before that, the common mode of transmission from person to person, the need to wear a mask, maintain social-distancing and other common practices were discussed.

  • What is the first thing the virus does when it enters the body?

  • How do the symptoms start developing?

After this, we came on to the binding of this virus on the ACE-2 (Angiotensin-converting enzyme 2) receptor.
ACE-2 receptor is meant for the binding of Angiotensin-converting enzyme.
Here, the COVID19 binds to this receptor.

  • How can a virus bind to the ACE-2 receptor which is meant for something else?
    The virus has a protein coat which consists of spike proteins (like the one denoted in the above image)
  • Is the virus smart :smirk: enough to change the structure of its protein to bind to the receptor accordingly?
    It can be thought of as a chance event that the protein structure of the virus and the shape of the receptor is such that both are compatible with each other!
    (A chance event has lead to a pandemic!!)

Here is where the drama starts.

The SARS-CoV had occurred in 2003, this too is a coronavirus of the same ancestry of COVID19. But, back then it had affected less than 10K people and now, its novel strain has affected more than 1Crore people worldwide.
(MERS-CoV had another outbreak in 2012)

  • Why this? The same ancestry, similar symptoms but lethality…!?

  • What makes/is making all the three strains of this virus different from each other?

We made a proposition/hypothesis (which seemed logical, may/may not be true) that the SARS-CoV had a circular protein coat back then (as shown in the diagram above) and this COVID19 or SARS-CoV2 has spike proteins.

  • How is this helping it in affecting the host effectively?
    Evolution? Mutation? How?

These keywords made us discuss the things on the left-hand side in the above photograph.

I hope this shall trigger a discussion. Let’s not leave it.

"Nothing in Biology Makes Sense Except in the Light of Evolution"
~ Theodosius Dobzhansky

Interesting, Eh?


16/9/20in today’s discussion we talked about Phyllanthus plant that opens up its leaves during the sunrise time and closes its leaf around the sunset time showing a sleep-wake pattern like humans are active during the day and sleep during the night where the opening of the leaf is indicating awakeness and closing of the leaf is showing sleep of the plant and hence the word sleep-wake cycle and this cycle is repeated every day so we started comparing the sleep-wake pattern of a plant from different parts of the country because the sunrise and sunset time would be varied in various parts of the country so is it that this particular time that the leaf is closing or opening or is it due to sunrise or sunset then we talked about the internal clock is there any internal clock within the plant that is making the plant open or closed the leaf during a definite time or light is playing any role This internal clock could be independent of the sunlight
can we build any hypothesis and can we design an experiment based on our hypothesis finding out is light playing a role or is it such that the plant is opening up for 12 hours and closing for 12 hours

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we also discussed fruit flies. Whereby can we identify the fly, clearly by looking at some of the features All of us know it is an animal it belongs to kingdom Animalia. It belongs to Phylum Arthropoda [segmented body, bilateral symmetry-the left and right sides of an arthropod are the same – it will have the same number and arrangement of legs, eyes, etc. on the right side of its body as on the left.] (simple example butterfly, lobster, bee, cockroach). It belongs to class Insecta [Insects have three main body segments: the head, thorax and abdomen. They have six legs, one or two pairs of wings] (simple examples cockroach, butterfly, fruit fly, house fly, mosquito). It belongs to the order Diptera [ having two wings] (example s housefly and mosquito). classification could be done by clicking a photograph of the fly or viewing it, of course, We should not only look at the features of the fruit fly but also compare it with other organisms to say whether they belong to this family /order so this can be done at home next comes the family level. One of the short term objectives is to find Drosophila melanogaster from the native so for identifying the fly belonging to Drosophilidae family the wing patterns is to seen so the fly needs to be still the wing is to be magnified so usually when there was no lockdown we had access to the Labs /equipment like microscope were used to magnify the wings of the fly these equipment are not available at home, The idea was to develop our phone camera as a substitute for microscope can we get a good photograph and identify the fly.

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Aswathy and Siddhi have already tried some method for magnifying the wing of the fly Sidhhi tried using the torchlight with a transparent sheet on top of the torch and then placing the wing On It and click photography Aswathy has tried several methods that include the torchlight with 2 sheets of white paper and a transparent sheet with the wing only magnification is not enough to get a good picture resolution is important to get a clear image so that we can zoom the picture and see the pattern on the wing to identify the family of the fly

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In today’s discussion, we talked about photography and how it helps us identify any organism? Abhijeet from Mumbai had sent a photograph of a plant which closes its leaf during night time and opens during the daytime. Is it Phyllanthus plant how do we identify it? surely by looking at characteristic features of this Phyllanthus plant is which has a simple leaf whereas in the photograph that was shared by Abhijeet shows compound leaf particular Palmately compound leaf.WhatsApp Image 2020-09-17 at 1.16.50 AM

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what is palmately or pinnately compound leaf? for palmately compound leaf the leaflets form and radiate from a single point of attachment called the distal end another way to describe the palmate form is that the whole leaf structure is Palm where the leaves are projecting the leaf where are pinnate forms the leaves are not attached to the single point, for example, curry leaves the leaves are certainly not attached on a single point where is in Papaya leaf the leaves are attached to a single point So curry leaves is pinnately compound leaf whereas Papaya leaf is a palmately compound leafWhatsApp Image 2020-09-17 at 1.22.12 AM (1) WhatsApp Image 2020-09-17 at 1.22.12 AM

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Firstly Phyllanthus plant came under the family Euphorbiaceae but now it has a separate family called Phyllanthaceae that could be several reasons for it as to why the difference is been made. Why it has given a different family all together? one of the reason is that Euphorbiaceae plants are grown on land and one of the Phyllanthus species is aquatic.WhatsApp Image 2020-09-17 at 1.26.37 AM

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Nidhi had come across the similar plant of which Abhijit shared a photograph. She wanted to identify whether this plant is Phyllanthus or not and because Phyllanthus belongs to the Euphorbiaceae family. The characteristics of Euphorbiaceae family was the leaf should be alternate, opposite, whorled, simple or rarely palmately compound, stipulate whereas this plant had pinnately compound leaf that’s how she identified that this plant does not belong to Euphorbiaceae family and hence it is not Phyllanthus. How do we identify this particular plant? now some key features could be seen through the flower which was not seen in this picture hence the identification on the family level is a bit delayed.

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talking about Cardamine plants, Nazish wants to find out if we can germinate and grow Cardamine seed in kanamycin medium. she wants to design. question - does Cardamine grow in kanamycin? what will be the rate of germination of Cardamine seed in kanamycin? the expectation is to get the same rate of germination as in the normal tap water medium that is 100% in 10 days all the Cardamine seeds will germinate but not grow for this she is referring a paper on floral dip method in arabidopsis plant. as weighing machine is not available at home that is why we are making a stock solution of 1 mg per ml

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two cups are used for each set
10 seeds in each cup
each setup has one replicate
there are a total of 4 setups
30ml of setup solution +10 seed (submerged) in each cup [x2 per set up] {4 set up x 2= 8 test cups}
control- 30 ml of water + 10seed per cup [x2]

the stock solution
1000mg kanamycin powder in 1000ml of water

first set up solution - 2.5 per cent kanamycin
2.5ml of stock solution +97.5 ml of water=100ml

the second set up solution- 5 per cent kanamycin
5ml of stock solution + 95ml of water=100ml

third set up solution- 7.5 per cent kanamycin
7.5ml of stock solution +92.5ml of water=100ml

fourth set up solution -10 per cent kanamycin
10ml of stock solution +90ml of water=100ml

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