RP06

Calico, Green, Prickly

This page serves as a repository of research notes on Toxin RP06, the sixth mutagen unlocked in River Plains by Renard. It uses 3 Calico, Green, and Prickly (Negative to Reticulated,Spongy,Spotted) moss 2 duelling serpent,2 dung root, 4 heaven's torrent, 2 pool of tranquility, 3 spiderling mushrooms.

First mutation test: L-Vampire R-Silken

First mutation

The effect on L-Vampire is subtle, slightly bluer than a pure Vampire. Perhaps an IY was disrupted?

The effect on R-Silken is obvious, Cyan1 on the Outer and Middle stamen, requiring an IRR genome.

Only a replacement of the first gene (R) with an I would cause this effect, so RP06 hits the 1/18 of the Right Genome.

Additional information can be found from the second mutation test, which showed that RP06 hits 4/18 of the left Genome. The closest only I on Vampire this range would possibly hit would be gene 3/12.

Second mutation test: L-Silken R-Vampire

Second mutation

The effect on L-Silken is obvious, Magenta1 on the Outer and Middle stamen, requiring an IOO genome.

Only a replacement of the fourth gene (O) with an I would cause this effect, so RP06 hits the 4/18 of the Left Genome.

The effect on R-Vampire is subtle, but clearly an I was hit. Given the information from the first test, it must have hit 1/12.

Next step

Already I've been able to narrow down the gene hit locations quite well, but I'd like to get more precise with a larger genome. Clarity has a generous genome length of 36, and lots of Stamen related genes (I) toward the beginning of the genome. The mutagen should hit somewhere between the 6th and the 10th gene on Clarity.

The first 10 genes are as follows: IRIRIOIOIY

I will attempt to add an R into this sequence. If the R is added to the 6th or 8th gene, the outer stamen will become sea green. If the mutagen hits the 7th gene, an IO sequence will be disrupted, removing some magenta colour from the outer stamen creating a pale green. If the mutagen hits the 9th gene, an IY will be disrupted, removing some yellow colour from the outer stamen creating a bluish grey. If the mutagen hits the 10th gene, the IY will become and IR creating a slate blue colour.

For this to work, I need to find a lily with an R gene at the very beginning of the sequence. Choices include Fracture or Silken Fracture will do nicely, it is 18 genes in length so I know for certain what it will hit since its length matches Silken.

If the fourth test, were be Fracture Left, Clarity Right.

The fourth gene of Fracture, a G will be placed on either the first or second gene of Clairty. Either will look like the same result, which is not desirable.

The fourth test, will be Silken Left, Clarity Right.

The fourth gene of Silken, an O will be placed on either the first or second gene of Clarity. Visually, if the first or third gene is hit, the outer stamen will become a rosy brown. If the second or fourth gene is hit, the middle stamen will be a red-violet since instead of just disrupting an IR, the IR will become an IO.

With such distinctive results on a higher length genome, the target left and right genes should be narrowed significantly.

Third mutation test: L-Clarity R-Silken

Third mutation

An I was transplanted from the Clarity to the Silken and an IO allele was disrupted on the Clarity. Given our previous results, gene 7 must have been hit by the mutagen.

Fourth mutation test: L-Silken R-Clarity

Fourth mutation

No colour is visible on the Silken Stamen, so when the fourth gene was replaced, it was not replaced by an I narrowing down the possible genes to the second or fourth gene(R). The red-violet color on the stamen now present bears out this theory, as an IR allele must have been replaced by an IO. Almost certainly the second because if it affected the fourth out of 36 genes in this case, it would have affected the second out of 18 genes in Silken so I will rule out gene 4 based on that fact.

Next step

So, where to go from here. Ultimately I would like to characterize this mutagen based on Dusk, since that is the sea lily with the longest genome which is definitively known. On the right splint, this mutagen would hit genes 3-6.

The first eight genes of dusk are: GROR GORR, a magenta north and cyan north.

I'd like to test this against a new flower if possible, like Morning. On the left splint, this mutagen would hit genes 8-9. The first twelve genomes of morning are: IRRRIYYYIYYY

Eight possibilities exist:

L8R3

Dusk becomes: GRYR - a null set which will be visible via a loss in magenta to the north petals and no other effect

Morn becomes: IYYO which will be visible on the stamen by less yellow being visible in the inner stamen. This occurs with all L8 sets.

L8R4

Dusk becomes: GROY GORR - ROYG is a giant set which will change flower size, along with the loss of magenta to the north petals

L8R5

Dusk becomes: GROR YORR - making the cyan a null set

L8R6

Dusk becomes: GROR GYRR - again making cyan a null set, indistinguishable from L8R5.

L9R3

Dusk becomes: GRIR - Breaking the magenta north and visibly changing the pigment of the outer stamen.

Morn becomes: OYYY - A null set so color is removed frrm the stamen with no other effect

L9R4

Dusk becomes: GROI GORR - Breaking the magenta north and having no other effect.

Morn becomes: RYYY - A null set so color is removed from the stamen with no other effect

L9R5

Dusk becomes IORR - Breaking the cyan north, and coloring the outer stamen.

Morn becomes: GYYY - Intensifying yellow in the South petal set.

L9R6

Dusk becomes: GIRR - Breaking the cyan north and increasing cyan color in the outer and middle stamen.

Morn becomes: OYYY - A null set so color is removed from the stamen with no other effect

Of these, only L8R5 and L8R6 are indistinguishable. 25% chance of happening, not bad odds and the result on dusk can be readily clarified with additional tests.

What will happen with Dusk Left, Morning Right?

On the left, the mutagen will hit Dusk genes 13-16, a GYOR set.

On the right, the mutagen will hit Morning 2-3, either one an R from IRRR.

Again, 8 possibilities.

L13R2 or L13R3

Dusk becomes RYOR, breaking the dwarf set and having no other visible effect.

R2: IGRR - cyan color removed form all stamen

R3: IRGR - cyan color removed from the inner and middle stamen.

L14R2 or L14Re

Dusk becomes GROR, breaking dwarf and adding Magenta north.

R2: IYRR - cyan color removed from all stamen, yellow added to the outer stamen.

R3: IRYR - cyan color removed from the inner and middle stamen.

L15R2 or L15R3

Dusk becomes GYRR, breaking the dwarf set and having no other visible effect.

R2: IORR - cyan color removed from all stamen, magenta added to the outer stamen.

R3: IROR - cyan color removed from the inner and middle stamen.

L16R2 or L16R3

Swaps an R for R, perfectly ordinary Dusk and Morning.

This is more troublesome. I cannot distinguish between L13R3 and L15R3, nor can I distinguish between L16R2 and L16R3. Still, either result will provide an exact answer for either Dusk or Morning, and I might get a definitive answer for both.

Fifth mutation test: L-Morning R-Dusk

Fifth mutation

Little dusk is now normal size. This mutation was not expected to alter any dwarf genes, but one possible combination WAS expected to create a giant allele. Two giants and a dwarf combine to create a perfectly normal sized lily.

The inner stamen is really hard to see, but close examination shows that it is now cyan instead of green, indicating a loss of yellow. L8R4, the eight gene of Morning has mutated along with the fourth gene of Dusk.

Sixth mutation test: L-Dusk R-Morning

Sixth mutation

Aw, little dusk is all grown up now. The dwarf gene set has broken, leaving us with just a giant dusk and no other visible changes.

Cyan color has been removed from the inner and middle stamen of the Morning, leaving an ambiguous result for which dusk gene was hit, but we now know that the third gene of Morning was mutated.

Either the 13th or the 15th gene of Dusk was mutated, and further testing looks like it might be required, but wait! We learned something new from the fifth mutation test! When the RP06 Mutagen affected the eight gene of morning, that limits the upper found of the mutagen to affecting the 20.455% gene. Any higher and it would have hit the 9th gene instead (since 9/44 = .20455)! Since 15/73 = 20.548, it can't have been the 15th gene affected by the mutagen. Final result, L13R3.

Final tests

This leaves us with Blush (26), Crown (37), Delicate (16), Energy (42), and Fracture (18) yet to test. It is unnecessary to test fracture, with it's genome length of 18 it will behave like Silken in determining which genes are affected.

We know that the left splint range has been restricted to 16.667%-19.178%.

Blush: This should affect gene 5

Crown: This should affect gene 7

Delicate: This should affect gene 3

Energy: This should affect gene 7 or 8

We know that the right splint range has been restricted to 4.545%-6.849%.

Blush: This should affect gene 1

Crown: This should affect gene 2

Delicate: This should affect gene 1 This is unnecessary to test, since a larger genome has already been shown to affect the first gene.

Energy: This should affect gene 2 Solvents would be necessary to verify this anyways, so I won't bother.

There is some ambiguity about rounding so I'm not entirely sure about these results and will have to test them. Unfortunately, since the first five genes of Energy are V, it would either require revelation solvents or multiple tests to pin down exactly the results on the right splint.

6 flowers to test, wherever possible it would be best to test two unknown flowers in such a way that if a gene adjacent to the one I suspect is hit there will be an unambiguous sign.

Crown is expected to hit the seventh genome, an O.

This first 8 genes are as follows: UROOUROO

If the sixth gene is hit, it remains an R and no change is evident.

If the seventh gene is hit, URRO is formed, which is a null set.

If the eight gene is hit, UROR is formed, changing Magenta to Yellow.

Can we find an R on a different splint?

We can test on a Crown right as well!

Crown/Crown test

On the left splint, 9 results might happen

L6R1 UROOUUOO Breaking 1 Magenta West so both left and right flowers are visually identical

L7R1 UROOURUO Breaking 1 Magenta West so both left and right flowers are visually identical

L8R1 UROOUROU Breaking 1 Magenta West so both left and right flowers are visually identical

L6R2 UROOUROO No change, both flowers identical

L7R2 UROOURRO Breaking 1 Magenta West

L8R2 UROOUROR Replacing 1 Magenta West with a Yellow West

L6R3 UROOUOOO Replacing 1 Magenta West with a Yellow East

L7R3 UROOUROO No change, both flowers identical

L8R3 UROOUROO No change, both flowers identical

On the right splint, 9 results might happen

L6R1 RROOUROO Breaking one Magenta West

L7R1 OROOUROO Breaking one Magenta West

L8R1 OROOUROO Breaking one Magenta West

L6R2 UROOUROO No change

L7R2 UOOOUROO Replacing one Magenta West with Magenta East

L8R2 UOOOUROO Replacing one Magenta West with Magenta East

L6R3 URROUROO Breaking one Magenta West

L7R3 UROOUROO No change

L8R3 UROOUROO No change

The expected result and two possible abberations have unique results. Three possible abberations uniquely identifies at least the left splint gene hit, while three leave both the left and right gene affected ambiguous.

Delicate Left / Blush Right

The mutagen should affect the first gene on Blush on the right splint, but I'd like to make sure it doesn't affect the second. Similarly it should affect the third gene on Delicate on the left splint, but I'd like to make sure it doesnt affect the second or fourth.

Six posibilities exist.

Delicate IOIO Any result will remove Magenta in the outer stamen

L2R1 IGIO

L2R2 IRIO Gain Cyan in outer stamen

L3R1 IOGO

L3R2 IORO

L4R1 IOIG

L4R2 IOIR Gain Cyan in outer stamen

Blush GROR Magenta North broken in all cases

L2R1 OROR No other effect

L2R2 GOOR No other effect

L3R1 IROR Cyan in outer stamen

L3R2 GIOR Magenta in outer stement

L4R1 OROR No other effect

L4R2 GOOR No other effect

L3R1 and L3R2 (which includes the anticipated result) are both uniquely identifiable. L2R2 and L4R2 are indistinguishable, but identify the gene affected on the right splint. L2R1 and L4R1 are indistinguishable, but identify the gene affected by the right splint.

That leaves Blush Left and Energy Left to test.

Blush Left: Should affect the fifth gene (V), but let's look at the first 9 genes: GRORVGOOO

A U in position 4 would break the GROR

A U in position 5 would have no visible effect

A U in position 6 would change GOOO to UOOO: South magenta become East magenta

Perfect, but I don't have a Right U to hit so this will not be as straightforward.

Blush Left / Clarity Right

Blush on the left: GRORVGOOO

L4R2: GRORVGOOO No visible change

L5R2: GRORRGOOO No visible change

L6R2: GRORVROOO Lost outer south

Clarity on the right: IRIR

L4R2: IRIR No change

L5R2: IVIR Lost cyan in outer stamen

L6R2: IGIR Lost cyan in outer stamen

Uniquely identifiable results.

Energy left / Vampire Right Energy left is expected to hit 7 or 8, both Ys. First 9 genes: VVVVV UYYY

By putting an I in that place, how far the yellow color extends into the stamen will be a visible change regadless of whether it hits the 6th, 7th, 8th, or 9th gene! A Vampire on the right splint donates an I gene when hit by this mutagen.

Seventh mutation test: L-Crown R-Crown

Seventh mutation

Everything corresponds to the L7R2 result as predicted above.

Eighth mutation test: L-Delicate R-Blush

Eighth mutation

Difficult to tell the difference between Magenta-4 and Cyan 1 Magenta 3

Consultation with experts however concludes that it is Magenta 4, so the outcome is L3R2. The Second gene on Blush was affected, not the first.

Ninth mutation test: L-Blush R-Clarity

Ninth mutation

L5R2, exactly as predicted. A V was transferred from Blush to Clarity, removing some color from the outer stamen of the Clarity.

Tenth mutation test: L-Energy R-Vampire

Tenth mutation

Yellow color in the outer stamen only. The eight gene of Energy must have been affected.

Eleventh mutation test: L-Crimson R-Crimson

The Crimson lily genome has recently been approved for testing, so this test will be a quick check on whether a mutation on two of those lilies can be readily interpreted.

Left splint should hit gene 18 or 19
Right splint should hit gene 5 or 6

First 22 genes: GYOR IOOO IOO IO IO IYYY IYY

On the left splint, the middle stamen appears to have lost some Yellow, but the outer stamen is unchanged.
This would be explained by:
Left-Crimson 18 (Y)
Right-Crimson 6 (O)

However, colors do not get any richer after four color sets, so
Left-Crimson 18 (Y)
Right-Crimson 5 (I)
can't be ruled out.

On the right splint, the whole stamen appears to have lost some Magenta.
This is also consistent with:
Left-Crimson 18 (Y)
Right-Crimson 5 (I)

References

Genomes for reference to genomes.

Flower_Genome_Theories for interpretation of gene sets.

Mutagen_Research_and_Recipes for information on Mutagen recipes, and location for results to be loaded to.