Tobacco Mosaic Virus (TMV) Testing @ Home In Minutes

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oscar169

oscar169

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Tobacco mosaic virus (TMV) is a positive-sense single stranded RNA virus that infects plants, especially tobacco and other members of the family Solanaceae. The infection causes characteristic patterns, such as "mosaic"-like mottling and discoloration on the leaves (hence the name). TMV was the first virus to ever be discovered. Although it was known from the late 19th century that an infectious disease was damaging tobacco crops, it was not until 1930 that the infectious agent was determined to be a virus.
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History[edit]
In 1886, Adolf Mayer first described the tobacco mosaic disease that could be transferred between plants, similar to bacterial infections.[1][2] In 1892, Dmitri Ivanovsky gave the first concrete evidence for the existence of a non-bacterial infectious agent, showing that infected sap remained infectious even after filtering through finest Chamberland filter candles.[2][3] Later, in 1903, Ivanovsky published a paper describing abnormal crystal intracellular inclusions in the host cells of the affected tobacco plants and argued the connection between these inclusions and the infectious agent.[4] However, Ivanovsky remained rather convinced, despite repeated failures to produce evidence, that the causal agent was an unculturable bacterium, too small to be retained on the employed Chamberland filters and to be detected in the light microscope. In 1898, Martinus Beijerinck independently replicated Ivanovsky's filtration experiments and then showed that the infectious agent was able to reproduce and multiply in the host cells of the tobacco plant.[2][5] Beijerinck coined the term of "virus" to indicate that the causal agent of tobacco mosaic disease was of non-bacterial nature. Tobacco mosaic virus was the first virus to be crystallized. It was achieved by Wendell Meredith Stanley in 1935 who also showed that TMV remains active even after crystallization.[2] For his work, he was awarded 1/3 of the Nobel Prize in Chemistry in 1946,[6] even though it was later shown some of his conclusions (in particular, that the crystals were pure protein, and assembled by autocatalysis) were incorrect.[7] The first electron microscopical images of TMV were made in 1939 by Gustav Kausche, Edgar Pfankuch and Helmut Ruska – the brother of Nobel Prize winner Ernst Ruska.[8] In 1955, Heinz Fraenkel-Conrat and Robley Williams showed that purified TMV RNA and its capsid (coat) protein assemble by themselves to functional viruses, indicating that this is the most stable structure (the one with the lowest free energy). The crystallographer Rosalind Franklin worked for Stanley for about a month at Berkeley, and later designed and built a model of TMV for the 1958 World's Fair at Brussels. In 1958, she speculated that the virus was hollow, not solid, and hypothesized that the RNA of TMV is single-stranded.[9] This conjecture was proven to be correct after her death and is now known to be the + strand.[10] The investigations of tobacco mosaic disease and subsequent discovery of its viral nature were instrumental in the establishment of the general concepts of virology.[2]
 
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Structure[edit]


Schematic model of TMV: 1. nucleic acid (RNA), 2. capsomer (protomer), 3. capsid
Tobacco mosaic virus coat protein

A monomeric unit of the tobacco mosaic virus coat protein.[11]
Identifiers
SymbolCP
Entrez1494073
UniProtP03579
Tobacco mosaic virus has a rod-like appearance. Its capsid is made from 2130 molecules of coat protein (see image to the left) and one molecule of genomic single strand RNA 6400 bases long. The coat protein self-assembles into the rod like helical structure (16.3 proteins per helix turn) around the RNA which forms a hairpin loop structure (see the electron micrograph above). The protein monomer consists of 158 amino acids which are assembled into four main alpha-helices, which are joined by a prominent loop proximal to the axis of the virion. Virions are ~300 nm in length and ~18 nm in diameter.[12] Negatively stained electron microphotographs show a distinct inner channel of ~4 nm. The RNA is located at a radius of ~6 nm and is protected from the action of cellular enzymes by the coat protein. There are three RNA nucleotides per protein monomer.[13] X-ray fiber diffraction structure of the intact virus was studied based on an electron density map at 3.6 Ă… resolution.[11]
Physicochemical properties[edit]
TMV is a thermostable virus. On a dried leaf, it can withstand up to 120 degrees Fahrenheit (50 °C) for 30 minutes.
TMV has an index of refraction of about 1.57.[14]
Disease cycle[edit]
TMV does not have a distinct over-wintering structure. Rather, it will over-winter in infected tobacco stalks and leaves in the soil, on the surface of contaminated seed (TMV can even survive in contaminated tobacco products for many years). With the direct contact with host plants through its vectors (normally insects such as aphids and leaf hoppers), TMV will go through the infection process and then the replication process.
Infection[edit]
After its multiplication, it enters the neighboring cells through plasmodesmata. For its smooth entry, TMV produces a 30 kDa movement protein called P30 which enlarge the plasmodesmata. TMV most likely moves from cell-to-cell as a complex of the RNA, P30, and replicase proteins.
It can also spread through phloem for longer distance movement within the plant. Moreover, TMV can be transmitted from one plant to another by direct contact. Although TMV does not have defined transmission vectors, the virus can be easily transmitted from the infected hosts to the healthy plants, by human handling.
Replication[edit]
Following entry into its host via mechanical inoculation, TMV uncoats itself to release its viral [+]RNA strand. As uncoating occurs, the MetHel:Pol gene is translated to make the capping enzyme MetHel and the RNA Polymerase. Then the viral genome will further replicate to produce multiple mRNAs via a [-]RNA intermediate primed by the tRNAHIS at the [+]RNA 3' end. The resulting mRNAs encode several proteins, including the coat protein and an RNA-dependent RNA polymerase (RdRp), as well as the movement protein. Thus TMV can replicate its own genome. After the coat protein and RNA genome of TMV have been synthesized, they spontaneously assemble into complete TMV virions in a highly organized process. The protomers come together to form disks or 'lockwashers' composed of two layers of protomers arranged in a helix. The helical capsid grows by the addition of protomers to the end of the rod. As the rod lengthens, the RNA passes through a channel in its center and forms a loop at the growing end. In this way the RNA can easily fit as a spiral into the interior of the helical capsid.[15]
 
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Host and symptoms[edit]


Tobacco mosaic virus symptoms on tobacco


Tobacco mosaic virus symptoms on orchid
Like other plant pathogenic viruses, TMV has a very wide host range and has different effects depending on the host being infected. The tobacco mosaic virus has been known to cause a production loss for flue cured tobacco of up to two percent in North Carolina.[16] It is known to infect members of nine plant families, and at least 125 individual species, including tobacco, tomato, pepper (all members of the useful Solanaceae), cucumbers, and a number of ornamental flowers.[17] There are many different strains. The first symptom of this virus disease is a light green coloration between the veins of young leaves. This is followed quickly by the development of a "mosaic" or mottled pattern of light and dark green areas in the leaves. Rugosity may also be seen where the infected plant leaves display small localized random wrinkles. These symptoms develop quickly and are more pronounced on younger leaves. Mosaic does not result in plant death, but if infection occurs early in the season, plants are stunted. Lower leaves are subjected to "mosaic burn" especially during periods of hot and dry weather. In these cases, large dead areas develop in the leaves. This constitutes one of the most destructive phases of tobacco mosaic virus infection. Infected leaves may be crinkled, puckered, or elongated. However, if TMV infects crops like grape and apple, it is almost symptomless.
Transference to humans[edit]
Consumption of tobacco products infected with the tobacco mosaic virus has not been found to have any effect on humans.[citation needed]
Environment[edit]
TMV is known as one of the most stable viruses. It has a very wide survival range. As long as the surrounding temperature remains below approximately 40 degrees Celsius, TMV can sustain its stable form. All it needs is a host to infect. If necessary, greenhouses and botanical gardens would provide the most favorable condition for TMV to spread out, due to the high population density of possible hosts and the constant temperature throughout the year.
Treatment and management[edit]
One of the common control methods for TMV is sanitation, which includes removing infected plants, and washing hands in between each planting. Crop rotation should also be employed to avoid infected soil/seed beds for at least two years. As for any plant disease, looking for resistant strains against TMV may also be advised. Furthermore, the cross protection method can be administered, where the stronger strain of TMV infection is inhibited by infecting the host plant with mild strain of TMV, similar to the effect of a vaccine.
In the past ten years, the application of genetic engineering on a host plant genome has been developed to allow the host plant to produce the TMV coat protein within their cells. It was hypothesized that the TMV genome will be re-coated rapidly upon entering the host cell, thus it prevents the initiation of TMV replication. Later it was found that the mechanism that protects the host from viral genome insertion is through gene silencing.[18]
Scientific and environmental impact[edit]


TMV virus: super resolution light microscopy
The large amount of literature about TMV and its choice for many pioneering investigations in structural biology (including X-ray diffraction), virus assembly and disassembly, and so on, are fundamentally due to the large quantities that can be obtained, plus the fact that it does not infect animals. After growing a few infected tobacco plants in a greenhouse and a few simple laboratory procedures, a scientist can easily produce several grams of virus. As a result of this, TMV can be treated almost as an organic chemical, rather than an infective agent.
Investigational uses[edit]
Due to its cylindrical high aspect ratio, self-assembling nature, and ability to incorporate metal coatings (nickel and cobalt) into its shell, TMV is an ideal candidate to be incorporated into battery electrodes[19] Addition of TMV to a battery electrode increases the reactive surface area by an order of magnitude, resulting in an increase in the battery's capacity by up to six times compared to a planar electrode geometry.[19][20]
 
oscar169

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So all of a sudden I had a few leafs start to look like the one pictured below, so I did a bunch of reading on the net about the TMV Virus, and found a lot of different info on it. Here is the step by step @ Home test that anyone of you can do, to find out if you have it or not....;)
Just have a clean surface, I used brand New Razors every test cleaned with 99% Rubbing Alcohol also wore new cloves for every test, keep everything SUPER CLEAN or you will Fuck up the Test..:stop:
Pic1TMV
Pic2TMV
Pic3TMV


Here are the pics of the Test Kit, everything must be stored in the Fridge at all times !!

IMG 1078

IMG 1077


IMG 1064

IMG 1067
IMG 1070
IMG 1072
IMG 1076
 
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Natural

Natural

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so that would be a positive result for TMV?
whatever my Chemdog D cut had transferred over to my white cut..at least from my perspective that's what happened.
 
Seamaiden

Seamaiden

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Y'all remember @Tobor the 8th Man? He submitted a bunch of samples to be tested for various mosaic viruses, and they all came back negative. I sure do miss him.
 
oscar169

oscar169

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Y'all remember @Tobor the 8th Man? He submitted a bunch of samples to be tested for various mosaic viruses, and they all came back negative. I sure do miss him.

Where did he send them ?
That's what is so nice about this test, you can do it at home, you get a new clone mother, Hell test her before you bring it into you gardens....;)
 
Seamaiden

Seamaiden

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That I couldn't say, I just remember that he did it and did a write-up on it for folks here at the farm.
 
BrianDirt

BrianDirt

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So whats wrong with the plant then Ph high? or missing a micro nute?
 
Skunkmasterflex

Skunkmasterflex

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Great thread and awesome info. I have a plant iv seen put off a leaf like that now and then. I personally think it's a deficiency of sorts. When the strain is dialed to hell it never happens but when I used to slack on feeding it silica or calmag it woukd pop up. I'd never getaa whole plant with leaves Like that, just one or two on the whole plant when it did happen.

Very interesting. We need some people with strong knowledge in both science and botany on here to tell us what's up. I'm sure a well educated botanist would know the problem.
 
justanotherbozo

justanotherbozo

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...can you by any chance share a link to where the test kits can be found?

tia, bozo
 
smokie

smokie

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In my white crosses that displayed this from Raskal it would seem to get darker or more visible with higher doses of nutes
 
leadsled

leadsled

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Tobacco mosaic virus (TMV) is a virus for tobacco! Not cannabis.

According to a Plant Pathologist. The cannabis plant is not going to get infected with tmv.


I agree with the above posters, Chemdog D, The white both show this trait if they are needing more food.
Crosses of the two can transfer the trait. I have 30+ mothers and it does not spread to other plants.


Have also heard a new "rumour" of the tmv being broad mite/cyclamen damage. :yuck:
 
Seamaiden

Seamaiden

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The question for me is this--what can infect cannabis? Isn't there a hemp streak virus? And then what about what lilmisslone found all those years ago, barley streak virus? (which didn't make sense to me that it could also infect cannabis, but viruses are clever little packets)

I think there may be other strains that will show this, then. I've seen it on my old cut of The White, my old Chemdog D cut, and ONE cut of Shut The Fuck Up, not all, just one. Not the mother, a clone. That would suggest that it very well be more like a nutritional effect, however, I cannot stop thinking about the Russian fox experiment, where they simply selected for one trait--friendliness--and the foxes' furs became unusable because they began showing what patches, much like domestic dogs.

Get this, we're seeing it now in captive bred clownfishes, in fact there's a 'platinum' variety that's popular now but wouldn't last a minute in the ocean. So, perhaps this is another side effect of domestication, too.
 

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