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Author Topic: Pollination basics to advanced....Pollen storage tools/grafting tools  (Read 14972 times)

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Offline Inyan

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http://www.orchardvalleysupply.com/ovsstore/pc/Parafilm-Grafting-Tape-p278.htm
Grafting tape that stretches and breathes ideal for seedling grafts to full size specimen grafting.
http://www.argos-tech.com/c-3-p-57-id-3.html

1.5 ml black centrifuge tubes for pollen storage. Dry pollen prior to storage for 1-2 hours and then freeze. The lower the temperature the longer the viability of the pollen.
Wracks for storing pollen http://www.phenixresearch.com/pc_combined_results.asp?search_cat=searchexact~pcpcm.parent_pc_id~78119AE48C934DD4A68D47130EC32B5B&pc_id=78119AE48C934DD4A68D47130EC32B5B
Remember, properly label all centrifuge tubes and or wracks prior to storage. Memory is not to be relied on an only take one type of pollen out at a time.

The sites I listed are just for example to show you what types of supplies works well for me. They are not necessarily the best prices to be found, but they do work. Anyone interested in making hybrids, selecting for new cultivars out of a regional variety, or otherwise seeking to improve the genetics or genetic variability of their collection may find these tools or tools similar to these essential for their progress as we may not always have the flowers we want in bloom at the moment another flower is receptive. Storing pollen properly and shipping pollen are all methods the top hybridizers, breeders, cultivators, etc. rely on. Pollen that is shipped should be shipped in a container, such as a Styrofoam container, with sufficient cold packs to keep pollen at 2-8 degrees C. Any pollen shipped should ideally be used asap or refrozen at colder temperatures to ensure continued viability of pollen. It is not unusual to have lowered germination rates with pollen that has been frozen and then refrozen so when possible replenish pollen stores or use just once after bringing pollen out of freezer. If the pollen is rare or super valued plan your crosses accordingly or transfer some of the pollen to another medium for transfer to the stigma so as not to warm up the entire collection of pollen. You must also be ever vigilant for the possibility of cross contamination with collected pollen. Pollen stored on fluffed up q-tip heads for example... once used are best not used again as they may acquire  pollen from the cacti they were used to pollinate. Emasculate the flower when possible where the cacti is self pollinating. The emasculation process is ideally done before the anthers produce pollen and can often be done before the flower opens up. Furthermore, pollination can also many times be done before the flower opens up as the pistil may be receptive long before it has matured with many species. If the pollen itself is preprogrammed to terminate at a shorter distance... i.e. the pollen was collected from a species in which the pistal was comparably shorter than the cacti it is supposed to pollinate then one may need to account for this and pollinate before the pistal has matured and grown longer, perform a cut style pollination, or simply try and hope for the best. Experimentation is key as well as never assuming that partial fertility or infertility is the same thing as totally infertile. There are many variables to pollen incompatibility just as there are many types/forms geometric, diploid, tetraploid, etc. to be found in sometimes rare and unlikely numbers... but possible nonetheless if sufficient attempts are made.
 

For those that graft...
Every areole is a cactus waiting to be born

Offline Inyan

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Re: Pollination basics to advanced....Pollen storage tools/grafting tools
« Reply #1 on: January 27, 2015, 04:44:42 PM »
L.Williamsii has been successfully fertilized by Ariocarpus Fissuratus, L.Diffusa, Mammillaria Bocasana, M.Zeilmanniana, Strombcactus Disciformis, Turbinicarpus Pseudomacrochele, Astrophytum Asterias, and Epithelantha Micromeris.

http://www.magicactus.com/description.html

What is not said in the above document is whether the self-fertile regional varieties of Lophophora were used in these crosses or whether the self incompatible Lophophora williamsii regional varieties were used in the crosses. This may make a huge difference. Also of importance is the fact that L. williamsii is listed as the mother plant in these crosses. When using a self-fertile variety as a seed bearer it is important to first emasculate the flower to remove anthers and pollen before the pollen ripens to prevent self pollination. It is also important to protect said cacti from other sources of pollination besides the one you are attempting to make. With that said, sometimes the use of mentor pollen or pollen cocktails may need to be utilized to get distant crosses to take. Again, I was not able to find mention of specific techniques utilized in the pollination attempts themselves. Suffice it to say that many of these crosses if carried out over more than one generation could yield interesting cacti of sufficiently different outward appearance as to make them indistinguishable from any pure species Lophophora williamsii and with several generations of crossing siblings backcrossing one could introduce a stable cultivar with these new traits that was completely fertile both ways with the pure species Lophophora williamsii in theory. Food for thought at any rate for those interested in going that route. Seeing a more brilliantly red colored flower with strong kikko koh-yo traits on a young L. williamsii would indeed be interesting, but perhaps a very long project indeed.
For those that graft...
Every areole is a cactus waiting to be born

Offline Chief BigTittyFlapFlaps

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Re: Pollination basics to advanced....Pollen storage tools/grafting tools
« Reply #2 on: January 28, 2015, 12:16:28 AM »
how would you go about breeding for cold hardiness?  i don't know how to go about this other than killing a bunch of big bends and breeding the survivors over and over. 

Offline Inyan

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Re: Pollination basics to advanced....Pollen storage tools/grafting tools
« Reply #3 on: January 28, 2015, 09:57:10 PM »
Breeding for any trait can be quite an undertaking depending on how one wants to go about it. If the trait is already in your selection then it is a simple matter of breeding back to your one cacti that has the trait. This would be your f1 and as such pretty useless in most cases other than to breed again back to your one special cactus that has the trait or one could breed siblings together and this would be your f2 or filial generation 2. However, it does get tricky as sometimes traits are only passed via pollen donor or seed bearer. This means you have to keep track of how your cross was made and do the cross both ways for optimum success. Mother plant goes first in a cross and pollen donor second. So, your cold hardy cross would be Cold hardy cacti 1 (mother) x regular 2 (pollen donor). So f1 = CHC1 x R2, but this doesn't guarantee those traits will even be passed on. You need to do another cross also f1 = R2 (seed bearer) x CHC1 (pollen donor). Now, your f1 selections generally won't be worth much. Your f2 however (CHC1 x R2) x CHC1R2 siblings however should throw some interesting possibilities your way. Your f2 cross is always where you have to grow out the most seedlings for trial and testing and depending on gene combinations... you may have to grow out a great many to get some improvement. Your f2 should also be done (R2 x CHC1) x (R2 x CHC1) and another f2 (R2 x CHC1) x (CHC1 x R2) and yet another f2 of (CHC1 x R2) x (R2 x CHC1) and so on. Only in this manner will you get the full potential by growing out several hundred of seeds from a single cross and still you may not have tapped the full potential depending on how many variables you are working towards. Now, selective pressures may be given at the f2 or f3 seedling stage to begin to select for cold hardiness by dropping the temperature down to the lower range of known cold hardiness. You have to do this gradually and for limited amounts of time and select out those few that survive without killing them all off. Now, ideally you may already have a regional selection to work from that shows the characteristics you are after, but if not you may have to involve other known species or a different genus altogether that has some compatibility and work this into your plan. This is still complicated however as your goal is simply for cold hardiness. This means that you will then have to breed this trait in from an outside genus or species and then breed out all those traits you do not desire to arrive back at your pure species form. Purists may argue that you will never arrive at a pure specimen, but such is the nature of their argument... they fail to understand evolution, natural selection, or man-made selections. Next, you have to learn to graft if your interest is serious as grafting speeds up your selection process as well as your ability to set seed. Remember, species evolved slowly over time and you are attempting to guide that selection in a much faster time frame based on your needs and desires. I hope this general framework proves helpful for you. The most important thing to remember is that your only as good as the seeds and cacti you have to work from. With that said, it is best to buy the best you can afford and start from their rather than to start from a less than ideal place and waste years and time. Genetic diversity is key when looking for unknown traits or trying to breed something new, but then you also have to look at regional variations and diversity. Perhaps there is some gene hidden where it is not needed in a southern local that when mixed with a northern local may have interesting results? The bottom line is that you are often working with unknowns and that is fine. Try to work from as many known values as you can however and introduce those unknowns as needed. That would be my guidance. That and never assume just because something hasn't been done or doesn't look like it can be done doesn't mean it can't be done. Just that it hasn't been figured out yet or the right technique has not been tried yet. Suffice it to say that the quickest way to get a cold hardy L. williamsii that could grow in Ontario for instance may be protoplast fusion with Opuntia compressa. Double the chromosomes and then select out for the most potent seedlings and so and so forth. Yes, that kind of expertise is beyond many and perhaps I should not have even brought it up. I tend to get wordy at times. But for me the excitement is in the chase. I look at some strains like Super Silver, Black Widow, AK47, and the like and I wonder... what could be done with more people working towards any goals whatsoever with cacti like L. williamsii, Trichocereus, etc. L. williamsii will breed with quite a few different cacti and yet we do not see any great improvements being made on the whole nor do we see even great improvements being made even with simple L. williamsii (southern) x L. williamsii (northern) regional variations. There is so much that can be done if there were more people interested in the genetics, growing, and testing of these specimens. True, a L. williamsii that was 10 times as potent as a normal field collected specimen may beg several generations in the making and might require that only a single specimen be utilized in a ceremony for instance... these are places I think could bear serious exploration. A few people are working toward more potent and larger varieties of Trichocereus. I would love to see colorful flowers added to that agenda and I think that as more people get involved in hybridizing we will see new and interesting forms arise. Variegate, crested, monstrose forms will become more common and there will be a sea of better hybrids out there. The danger I do see in these forms are that some of them may not be as easily identifiable as their former pure species selves. Some, may prove to be so potent that those that partake of them may over indulge whether it is in a ceremonial context or a relaxing back yard adventure type context. I know there are many who partake of these sacred cacti for pleasure and many who partake of them for religious reasons. Still, others partake of them for deeper insight into their own minds. I think wherever one goes with their hybridizing agenda's it is wise to keep all these things in mind as real progress is made. When in doubt, it is better to risk too little than to risk too much. I think we are still a good ways away from the kind of progress I see as being possible, but the more people that get involved in hybridizing and the more people that share like minded goals the more interesting and potent varieties of cacti we are going to see. L. williamsii is one of those cacti that is typically strong enough that a single button is enough for many.

I remember one ceremony where a Mexican came to Sundance. He dragged 28 buffalo skulls and later went to a cross fire ceremony where he did not understand that he was not supposed to eat the pejuta every time it passed by. The next day, he sat smiling and drooling. So, context plays a big part in these things. Where your mind is at prior to and during the experience. Our Mexican friend did not know Lakota and he did not know English so there was only one person there who could interpret for him and he was unfortunately not their until after that experience. But, even so... the experience was such that it was positive for him. For anyone else... out of context I don't think the experience would have went so well.
For those that graft...
Every areole is a cactus waiting to be born

Offline Chief BigTittyFlapFlaps

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Re: Pollination basics to advanced....Pollen storage tools/grafting tools
« Reply #4 on: January 28, 2015, 11:56:32 PM »
your not wordy at all dude.  this is extremely helpful for me and anyone else who reads it.  sometimes it's hard to adequately respond because it's so much to soak in.  for cold hardiness i was expecting just to play it by chance.  i thought by germinating 1000's and killing off all but ten % with cold and then repeating the process would get me some results.  now I'm thinking there must be more factors involved like resistance to rot.  i would have to really sit down and plan this out, and then chart it all the way along.  i'm absolutely with you on the untapped hybridizing potential of lophophora.  i just don't know exactly what direction i want to take go.  aside from my regular lophs here's my starting point…

Offline Chief BigTittyFlapFlaps

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Re: Pollination basics to advanced....Pollen storage tools/grafting tools
« Reply #5 on: January 29, 2015, 12:06:23 AM »
-camargo
-texensis
-parras
-san jose
-cardona
-est catorce
-pentagona
-casa grande
-ramos arrizpe
-el nunez
-eintrogue

i will also be getting some huizache and big bends and that will be my starting point.  any info is greatly appreciated.  i can't find out much info on some of these. 

Offline Chief BigTittyFlapFlaps

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Re: Pollination basics to advanced....Pollen storage tools/grafting tools
« Reply #6 on: January 29, 2015, 12:06:59 AM »
.

Offline Inyan

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Re: Pollination basics to advanced....Pollen storage tools/grafting tools
« Reply #7 on: January 29, 2015, 10:04:16 PM »
Your texensis is a northern type and so should be self-fertile. This means you will want to use it as a pollen parent to your southern types that are not self-fertile such as Est Catorce, Pentagona, Huizache, etc. if your goal is to increase both genetic diversity and hardiness. I say this as texensis while it has some admirable characteristics and is most notably a northern growing regional variety... is also not as genetically diverse as it is self-fertile. So, ... I would seek to outcross your self-fertile varieties with your more distantly related specimens that are not self-fertile to initially increase the genetic diversity of your seedling pool. These f1 specimens from each cross can then be bred systematically to each other and back to their original parents to uncover more traits of interest. It is the F2 generation where you may start to see some interesting results. Any traits you uncover that are of interest should be selected for by making those specimens your primary pollinators with the understanding that the trait may not be carried by the pollen at all. This means you may have to make the reverse cross or simply take your two best specimens showing that particular trait and breed them to each other. If the traits seemingly disappear you may have those traits simply hidden in your next f1 selection and to uncover them you will have to breed again back to your original selections. Many hybridizers simply assign numbers and or letters to their many seedlings to help keep track of their crosses.... Good tools one may find to use to help visualize the genetics of inheritance exist on the web that will actually calculate phenotypes and genotypes out to 5 different traits and more exist that can be paid for with more detailed information able to be stored.
This is an easy free source for looking at up to five traits. http://scienceprimer.com/punnett-square-calculator
« Last Edit: January 29, 2015, 10:09:26 PM by Inyan »
For those that graft...
Every areole is a cactus waiting to be born

Offline Chief BigTittyFlapFlaps

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Re: Pollination basics to advanced....Pollen storage tools/grafting tools
« Reply #8 on: January 29, 2015, 11:03:55 PM »
alright that gives me an idea to get started.  all the seeds are down now.  while their growing i need to focus my efforts on figuring out how to graft. 

Offline Inyan

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Re: Pollination basics to advanced....Pollen storage tools/grafting tools
« Reply #9 on: January 29, 2015, 11:13:29 PM »
Your current attempt is by far the best you have done and I have high hopes that they both will take. Could it have been done better? Yes, but was it done well enough? The jury may still be out on that one, but I'd say you have a very high probability of those last two taking. Grafting with Opuntia is sometimes considered easier for another reason... you don't have to line up two circles or offset two growth rings as the Opuntia has a more horizontal or elliptical ring making it a bit easier to work with.
For those that graft...
Every areole is a cactus waiting to be born

Offline Chief BigTittyFlapFlaps

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Re: Pollination basics to advanced....Pollen storage tools/grafting tools
« Reply #10 on: January 30, 2015, 11:29:51 PM »
thanks inyan.  i have a pretty good feeling about these ones but the next ones i will be stripping the stem down a ways and using saran wrap.  that makes sense about the opuntias. will have to chop a piece just to have a look.  hey have you ever done any grafting with adeniums? 

Offline Inyan

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Re: Pollination basics to advanced....Pollen storage tools/grafting tools
« Reply #11 on: January 31, 2015, 05:33:32 PM »
I sincerely wish I had more pictures of some of my old grafts for you. I will be making some fresh grafts this year and will post on this forum. For now, this is a picture of one of my seedling grafts I did a few years back. Cacti are excellent to begin to learn how to graft with as they are so forgiving. This particular graft in the picture is a seedling of a Brugmansia I grafted just after it sprouted. As you can see, the seedling is on the left and has had its roots removed already so it is totally dependent on the mother plant for its nutrients. Suffice it to say that Brugmansia grow very rapidly from seed and this grafting was really not needed to speed it up growth wise. I was experimenting with other variables that I was interested in however such as earlier bloom time, cross compatibility issues, etc. Any material you can find that you can work with without fear of losing its genetic diversity is a plus when learning to graft.
« Last Edit: January 31, 2015, 05:37:20 PM by Inyan »
For those that graft...
Every areole is a cactus waiting to be born

Offline Inyan

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Re: Pollination basics to advanced....Pollen storage tools/grafting tools
« Reply #12 on: January 31, 2015, 05:58:30 PM »
Some of these grafts are a little sloppy, but no worries here as the grafts always look better after they heal. Besides the obvious benefits of having different flowers on the same plant I was also interested in chimera formation with these grafts. I had too many other interests to list them all here without boring you to death. But here goes, I was interested in seeing if gene expression or regulation or compatibility could also be affected by grafting. I experimented with everything from cut style pollination, grafting ovaries, etc. for a time and suffice it to say I was only able to come up with some mature seed pods that should not have formed and even mature seeds... however... the internal embryo's were usually aborted early on and would have necessitated embryo rescue or possibly embryo grafting to attain anything of note.
For those that graft...
Every areole is a cactus waiting to be born

Offline Chief BigTittyFlapFlaps

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Re: Pollination basics to advanced....Pollen storage tools/grafting tools
« Reply #13 on: February 01, 2015, 01:31:03 AM »
very cool man!  i have these two different coloured adenium obesums i am trying to figure out what to do with.  i've seen some pretty cool artwork with these and i want to come up with something cool.  I've never grown brugs but grew datura purple ballerina the last few years.  the flowers smelled pretty amazing. 

Offline Inyan

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Re: Pollination basics to advanced....Pollen storage tools/grafting tools
« Reply #14 on: February 01, 2015, 01:00:16 PM »
I've never grafted Adenium, but from what I understand it is a little harder than grafting Brugmansia, but not much more. Grafting Datura is just slightly harder than grafting Brugmansia due to the fact that Brugmansia can be pushed much harder than Datura. If you like the fragrance from Datura metel... then you would be blown away by the fragrance of Brugmansia suaveolens or hybrids thereof. The nice part about Brugmansia is they can be stored away in the closet during the winter months and then brought back to life in the spring or summer. If you wish to improve your grafting skills, I would definitely consider very young Datura seedlings and any age Brugmansia. What you have to understand with Brugmansia however is that the flowering region where it forks should never be cut back to below the fork as that is where you the plant will flower. When making cuttings or grafts it is better to graft to a flowering portion of the Brugmansia if you wish to attain earlier flowers. The same can be said for cuttings. Make your cuttings in the flowering region and root those or graft those portions rather. Grafting can lend some peculiarities to your scion such as a dwarf root stock being used to dwarf the scion. Or a Pereskiopsis being used to accentuate the Kikko markings in Astrophytum. Some root stocks promote earlier flowering such as Opuntia compressa. There is much that can be learned first hand by using different stocks that may work better for you in your geographical area or under your growing conditions than it might work for someone else in a different area. This is why I find it important not to take everything you hear as gospel and experiment on your own when it comes to different grafting techniques, stocks, and even pollination methods. Old pollen that you might wish to discard as it has lost its fertility for example can be saved to use in pollen cocktails to trick a cacti into accepting a pollen it might not otherwise take or to increase the germination of those gametes etc. For example, I've been able to trick Brugmansia into accepting pollen and forming fully developed seed pods and even seed coats where it should not be possible using some of these methods. However, in my own attempts the embryo's inside the seed coats were either missing altogether at the macroscopic level or only partially formed. Early attempts to graft these embryo's failed on my part. However, this could be due to not enough practice ... i.e. poor technique. Or, there could still be the possibility that if I added more viable compatible pollen that this might push the growth hormones inherent in the seed pod itself to the point that even these embryo's that aborted early might be able to push a little further in their growth enabling a seed that while perhaps still immature... might be mature enough to germinate. Other techniques that should be considered to help with this process might be the doubling of the chromosomes using surflan by direct application to the flowering region to induce polyploidy of the gametes themselves. Still other techniques that should be considered are the production of tetraploid seeds by chemical means such as surflan using serial dilutions. Those seedlings that fail to germinate at different concentrations are gone, but those that germinate at the point just before this point may be considered strong candidates for tetraploid conversions. Those that are at the other end of the spectrum however can be most likely considered not affected by the conversion process. The other benefits to this conversion process are numerous... phenotype expression could be markedly changed and hybrids may be able to be formed with such treated seed that could not be possible otherwise. This process is exactly why we have some of the amazing Hemerocallis hybrids on the market today.
For those that graft...
Every areole is a cactus waiting to be born