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Thread: Pugz Nitro Express Reveg and Grafing Experiment.

  1. #11

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    Default Pugz Nitro Express Reveg and Grafing Experiment.

    I've just found this, it's going on to say about grafted plant swapping genetic material.

    So that means when a graft is successful the who plant changes it genetic make up,
    So in hein sight the buds should be different than the original ( before graft).

    Any way here it is

    For centuries, farmers have been genetically modifying their plants without even knowing it. That’s the message from German scientists who found that grafting, a common technique used to fuse parts of two plants together, causes the two halves to swap genes with each other.

    Grafting can involve fusing the stem of one plant (the scion) to the roots of another (the stock), or a dormant bud to another stem. There are many reasons for this – sometimes it’s the most cost-effective way of cultivating the scion, sometimes the stock has properties that the scion lacks including hardiness or sturdiness. The vessels of the two halves eventually merge but people have long believed that they keep their genetic material to themselves. It turns out they were wrong.

    Sandra Stegemann and Ralph Bock from the Max-Planck Institute tested the theory by grafting two strains of genetically engineered tobacco plant. A Samsun NN strain had its main genome loaded with a gene that produced a glowing yellow protein, and another that made the plant resistant to the antibiotic kanamycin. The second Petit Havana strain was engineered to produce a glowing green protein, and be resistant to spectinomycin, another antibiotic. These genes were shoved into the genome of its chloroplast, the small structures that allow plant cells to photosynthesise and that contain their own separate genetic material.

    Once the plants had merged, Stegemann and Bock found that the point of fusion was rife with cells that produced both glowing proteins and shrugged off both antibiotics. They cut slices from the plant and grew them in liquid that contained both kanamycin and spectinomycin for a month. While chunks that were taken from other parts of the plant fared poorly under these conditions, many of those from the graft site thrived, even producing fresh shoots.

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    You might think that these shoots survived because different cells were resisting the effects of the two antibiotics. But a glance under the microscope said otherwise. All the cells of the regenerating plants gave off a green fluorescence from their chloroplasts and a yellow one from their nucleus. This meant that every cell contained both proteins.

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    Stegemann and Bock even showed that the cells at the graft site were activating all of the added genes. They used a technique called Northern blotting, which detects whether genes are being switched on by recognising the RNA molecules that are produced during this process. First, all the RNA from the cells is extracted and separated according to their size. They are mixed with mildly radioactive probes designed to recognise specific RNA sequences – these give away the position (and size) of any RNA molecules of interest.

    Using these probes, the duo confirmed that all four of the inserted genes were being switched on in the fused plants. The new shoots can be bred with other plants to produce viable offspring, sowing that the swapped genes can be passed on through the generations.

    These results show that contrary to popular belief, grafted plants aren’t genetically distinct. As their tissues combine, so too do they swap genes with one another. Judging by the number of shoots that Stegemann and Bock produced, it’s a common event, but also a local one. The plants don’t exchange any genes over long distances.

    The number of chromosomes in these cells showed that the cells from the two plants hadn’t merged. Instead, Stegemann and Bock found that they were shuffling chloroplast DNA from one cell to another. Their chloroplasts of Petit Havana tobacco contain a distinct DNA sequence that’s unique to this strain, and that lies some distance away from the place where the extra genes were inserted.

    This marker sequence turned up in all the cells where genes had been swapped, which means that the plants were trading in very large chunks of DNA, or even their entire chloroplast genomes. The genes from the main nuclear genome, however, showed no signs of movement.

    Stegemann and Bock say that their discovery “further blurs the boundary between natural gene transfer and genetic engineering”. It also suggests that the trading of genes is more common than we thought, and occurs via routes we’re only starting to uncover. After all, plants can graft to each other without human help. The roots and branches of trees will sometimes merge when they touch one another. It’s a process called inosculation and it allows groups of trees to share nutrients and water through a combined root system. But trees of different species can also fuse together and in doing so, they may create a tunnel through which genes can pass.

    Reference: Stegemann, S., & Bock, R. (2009). Exchange of Genetic Material Between Cells in Plant Tissue Grafts Science, 324 (5927), 649-651 DOI: 10.1126/science.1170397

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  3. #12
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    Default Re: Pugz Nitro Express Reveg and Grafing Experiment.

    So the genes are swapped but they still keep there own genetic identity if I've read that right. So the trifle will keep its colours and the blue cheese will keep its smell but it could also pass these characteristics to the scion. Wonder if it could mix em all up together into the scion and produce cuttings with a mix of the lot lol. That would be cool.
    Last edited by mellowed; 09-03-13 at 06:31 PM.

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  5. #13

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    Default Pugz Nitro Express Reveg and Grafing Experiment.

    Some more info some of you may find helpful.




    Grafting and Budding Nursery Crop Plants

    When to Graft
    Unlike budding, which can be performed before or during the growing season, most grafting is done during winter and early spring while both scion and rootstock are still dormant. Containerized plants may be moved indoors during the actual grafting process; after grafting, these plants are placed in protected areas or in unheated overwintering houses. Field-grown stock, of course, must be grafted in place. Some deciduous trees are commonly grafted as bare rootstock during the winter and stored until spring planting. Indoor winter grafting is often referred to as bench grafting because it is accomplished at a bench.

    Selecting and Handling Scion Wood
    The best quality scion wood usually comes from shoots grown the previous season. Scions should be severed with sharp, clean shears or knives and placed immediately in moistened burlap or plastic bags. It is good practice during the harvesting of scions and the making of grafts to clean the cutting tools regularly. This may be done by flaming or immersing them in a sterilizing solution. Isopropyl (rubbing) alcohol also works well as a sterilant, although it evaporates quite readily. An alternative sterilizing solution may be prepared by mixing one part household bleach with nine parts water (by volume). However, this bleach solution can be highly corrosive to certain metals.
    For best results, harvest only as much scion wood as can be used for grafting during the same day. Select only healthy scion wood that is free from insect, disease, or winter damage. Be sure the stock plants are of good quality, healthy, and true to type. Scion wood that is frozen at harvest often knits more slowly and in lower percentage. If large quantities of scion wood must be harvested at one time, follow these steps:
    Cut all scions to a uniform length, keep their basal ends together, and tie them in bundles of known quantity (for example, 50 scions per bundle).
    Label them, recording the cultivar, date of harvest, and location of the stock plant.
    Wrap the base of the bundles in moistened burlap or sphagnum, place them in polyethylene or waterproof paper bags, and seal the bags.
    Store the bundles for short periods, if necessary, either iced down in insulated coolers or in a commercial storage unit at 32o to 34oF.
    Never store scions in refrigerated units where fruits or vegetables are currently kept or have been stored recently. Stored fruits and vegetables release ethylene gas, which can cause woody plant buds to abort, making the scions useless.
    Keep the scions from freezing during storage.

    NOTE: In grafting, as well as budding, the vascular cambium of the scion or bud must be aligned with the vascular cambium of rootstock. In woody plants the cambium is a very thin ribbon of actively dividing cells located just below the bark. The cambium produces conductive tissue for the actively growing plant (Figure 1). This vascular cambium initiates callus tissue at the graft and bud unions in addition to stimulating tissue growth on the basal ends of many vegetative cuttings before they have rooted.

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    Figure 1. Cross section of a woody plant stem.

    Types of Grafts
    Nurserymen can choose from a number of different types of grafts. This section describes only those basic types of grafts used on nursery crop plants.

    Cleft Graft
    One of the simplest and most popular forms of grafting, cleft grafting (Figure 2), is a method for top working both flowering and fruiting trees (apples, cherries, pears, and peaches) in order to change varieties. Cleft grafting is also used to propagate varieties of camellias that are difficult to root. This type of grafting is usually done during the winter and early spring while both scion and rootstock are still dormant. Cleft grafting may be performed on main stems or on lateral or scaffold branches.
    The rootstock used for cleft grafting should range from 1 to 4 inches in diameter and should be straight grained. The scion should be about 1/4 inch in diameter, straight, and long enough to have at least three buds. Scions that are between 6 and 8 inches long are usually the easiest to use.

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    Figure 2. Cleft graft.

    Preparing the Rootstock. The stock should be sawed off with a clean, smooth cut perpendicular to the main axis of the stem to be grafted. Using a clefting tool wedge and a mallet, make a split or "cleft" through the center of the stock and down 2 to 3 inches. Remove the clefting tool wedge and drive the pick end of the tool into the center of the newly made cleft so that the stock can be held open while inserting the scion.
    Preparing the Scion. In cleft grafting, one scion is usually inserted at each end of the cleft, so prepare two scions for each graft. Select scions that have three or four good buds. Using a sharp, clean grafting knife, start near the base of the lowest bud and make two opposing smooth-tapered cuts 1 to 2 inches long toward the basal end of the scion. Cut the side with the lowest bud slightly thicker than the opposite side. Be sure the basal end of the scion gradually tapers off along both sides.
    Inserting the Scion. Insert a scion on each end of the cleft, with the wider side of the wedge facing outward. The cambium of each scion should contact the cambium of the rootstock.
    Securing the Graft. Remove the clefting tool from the cleft so that the rootstock can close. Pressure from the rootstock will hold the scions in place. Thoroughly seal all cut surfaces with grafting wax or grafting paint to keep out water and prevent drying. If both scions in the cleft "take," one will usually grow more rapidly than the other. After the first growing season, choose the stronger scion and prune out the weaker.
    NOTE: The temperature of grafting wax is critical. It must be hot enough to flow but not so hot as to kill plant tissue. Recently, paint-like sealants have replaced wax in many areas because they are easier to use and require no heating.

    Bark Graft
    Bark grafting (Figure 3) is used primarily to top work flowering and fruiting trees. In contrast to cleft grafting, this technique can be applied to rootstock of larger diameter (4 to 12 inches) and is done during early spring when the bark slips easily from the wood but before major sap flow. The rootstock is severed with a sharp saw, leaving a clean cut as with cleft grafting.


    Figure 3. Bark graft.

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    Preparing the Stock. Start at the cut surface of the rootstock and make a vertical slit through the bark where each scion can be inserted (2 inches long and spaced 1 inch apart).
    Preparing the Scion. Since multiple scions are usually inserted around the cut surface of the rootstock, prepare several scions for each graft. Cut the base of each scion to a 1 ½- to 2-inch tapered wedge on one side only.
    Inserting the Scion. Loosen the bark slightly and insert the scion so that the wedge-shaped tapered surface of the scion is against the exposed wood under the flap of bark. Push the scion firmly down into place behind the flap of bark, replace the bark flap, and nail the scion in place by driving one or two wire brads through the bark and scion into the rootstock. Insert a scion every 3 to 4 inches around the cut perimeter of the rootstock.
    Securing the Graft. Seal all exposed surfaces with grafting wax or grafting paint. Once the scions have begun to grow, leave only the most vigorous one on each stub; prune out all the others. Bark grafts tend to form weak unions and therefore usually require staking or support during the first few years.

    Side-Veneer Graft
    At one time the side-veneer graft (Figure 4) was a popular technique for grafting varieties of camellias and rhododendrons that are difficult to root. Currently, it is the most popular way to graft conifers, especially those having a compact or dwarf form. Side-veneer grafting is usually done on potted rootstock.



    Figure 4. Side veneer graft

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    Preparing the Stock. Rootstock is grown in pots the season before grafting, allowed to go dormant, and then stored as with other container nursery stock. After exposure to cold weather for at least six weeks, the rootstock is brought into a cool greenhouse for a few days before grafting takes place to encourage renewed root growth. The plant should not be watered at this time.
    Make a shallow downward cut about 3/4 inch to 1 inch long at the base of the stem on the potted rootstock to expose a flap of bark with some wood still attached. Make an inward cut at the base so that the flap of bark and wood can be removed from the rootstock.
    Preparing the Scion. Choose a scion with a diameter the same as or slightly smaller than the rootstock. Make a sloping cut 3/4 to 1 inch long at the base of the scion. (Use the bark grafting technique shown in Figure 3.)
    Inserting the Scion. Insert the cut surface of the scion against the cut surface of the rootstock. Be certain that the cambia contact each other.
    Securing the Graft. Hold the scion in place using a rubber grafting strip, tape, or grafting twine. Seal the entire graft area with warm grafting wax or grafting paint. Remove the rubber or twine shortly after the union has healed. Never allow the binding material to girdle the stem.

    Splice Graft
    Splice grafting (Figure 5) is used to join a scion onto the stem of a rootstock or onto an intact rootpiece. This simple method is usually applied to herbaceous materials that callus or "knit" easily, or it is used on plants with a stem diameter of 1/2 inch or less. In splice grafting, both the stock and scion must be of the same diameter.


    Figure 5. Splice graft.

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    Preparing the Stock and Scion.Cut off the rootstock using a diagonal cut 3/4 to 1 inch long. Make the same type of cut at the base of the scion.
    Inserting the Scion. Fit the scion to the stock. Wrap this junction securely with a rubber grafting strip or twine.
    Securing the Graft. Seal the junction with grafting wax or grafting paint. Water rootstock sparingly until the graft knits. Over watering may cause sap to "drown" the scion. Be sure to remove the twine or strip as soon as the graft has healed.

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    Last edited by Pugz; 10-03-13 at 12:56 PM.

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  7. #14

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    Default Pugz Nitro Express Reveg and Grafing Experiment.

    Whip and Tongue Graft

    The whip and tongue technique (Figure 6) is most commonly used to graft nursery crops or woody ornamentals. Both the rootstock and scion should be of equal size and preferably no more than 1/2 inch in diameter. The technique is similar to splice grafting except that the whip on the rootstock holds the tongue of the scion in place (and vice versa). This leaves both hands free to wrap the joint.
    For the whip and tongue graft, make similar cuts on both the stock and scion. These cuts should be made with a single draw of the knife and should have a smooth surface so that the two can develop a good graft union. Up to this point, rootstock and scion are cut the same as for a splice graft.



    Figure 6. Whip and tongue graft.

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    Preparing the Stock and Scion. Cut off the stock using a diagonal cut. The cut should be four to five times longer than the diameter of the stock to be grafted. Make the same kind of cut at the base of the scion.
    Next, place the blade of the knife across the cut end of the stock, halfway between the bark and pith (on the upper part of the cut surface). Use a single knife stroke to draw the blade down at an angle through the wood and pith. Stop at the base of the initial diagonal cut. This second cut must not follow the grain of the wood but should run parallel to the first cut.
    Inserting the Scion. Prepare the scion in the same way. Fit the scion into the rootstock so that they interlock whip and tongue. Be certain that the cambia are aligned.
    Securing the Graft. Wrap the junction with a grafting strip or twine, and seal it with grafting wax or grafting paint. Never allow the binding material to girdle the stem.

    Saddle Graft
    Saddle grafting (Figure 7) is a relatively easy technique to learn and once mastered can be performed quite rapidly. The stock may be either field-grown or potted. Both rootstock and scion should be the same diameter. For best results, use saddle grafting on dormant stock in mid- to late winter. Stock should not be more than 1 inch in diameter.


    Figure 7. Saddle graft.

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    Preparing the Stock. Using two opposing upward strokes of the grafting knife, sever the top from the rootstock. The resulting cut should resemble an inverted V, with the surface of the cuts ranging from 1/2 to 1 inch long.
    Preparing the Scion. Now reverse the technique to prepare the base of the scion. These cuts on the rootstock and scion must be the same length and have the same slope so that a maximum amount of cambial tissue will make contact when the two halves are joined.
    Inserting the Scion. Place the V-notched scion onto the saddle of the rootstock. If rootstock and scion are the same diameter, cambial alignment is easier; otherwise adjust as needed.
    Securing the Graft. Wrap the graft with a grafting twine, tape, or strip, then seal it with grafting wax or grafting paint.

    All of the preceding techniques are used to top work horticultural crops for a particular purpose. Occasionally, however, grafting is used to repair injured or diseased plants. Two common techniques available for this purpose are bridge grafting and inarch grafting.

    Bridge Graft
    Bridge grafting (Figure 8) is used to "bridge" a diseased or damaged area of a plant, usually at or near the base of the trunk. Such damage commonly results from contact with grading or lawn maintenance equipment, or it may be caused by rodents, cold temperatures, or disease organisms. The bridge graft provides support as well as a pipeline that allows water and nutrients to move across the damaged area.
    Bridge grafts are usually done in early spring just before active plant growth begins. They may be performed any time the bark on the injured plant "slips."


    Figure 8. Bridge graft.

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    Preparing the Scion. Select scions that are straight and about twice as long as the damaged area to be bridged. Make a 1 1/2- to 2-inch-long tapered cut on the same plane at each end of the scion.
    Preparing the Stock. Remove any damaged tissue so the graft is on healthy stems. Cut a flap in the bark on the rootstock the same width as the scion and below the injury to be repaired. Gently fold the flap away from the stock, being careful not to tear the bark flap.
    Inserting the Scion. First, insert and secure the scion below the injury; push the scion under the flap with the cut portion of the scion against the wood of the injured stem or trunk. Then go back and insert and secure the scion above the injury following these same steps. Push the scion firmly into place. Pull the flap over the scion and tack it into place as described for bark grafting (Figure 3).
    When grafting with young stems that may waver in the wind, insert the scions so that they bow outward slightly. Bridge grafts should be spaced about 3 to 4 inches apart across the damaged area.
    Securing the Graft. Secure all graft areas with warm grafting wax or grafting paint. During and after the healing period, remove any buds or shoots that develop on the scions.

    Inarch Graft

    Inarching, like bridge grafting, is used to bypass or support a damaged or weakened area of a plant stem (Figure 9). Unlike bridge grafting, the scion can be an existing shoot, sucker, or watersprout that is already growing below and extending above the injury. The scion may also be a shoot of the same species as the injured plant growing on its own root system next to the main trunk of the damaged tree. With the inarching technique, the tip of the scion is grafted in above the injury using the same method as for bark or bridge grafting.


    Figure 9. Inarch graft.

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  9. #15

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    Interesting stuff pugz

    Can help a lot with people willing to try it out

    Maybe make a sticky for it in the advanced growing techniques section?

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    Default Pugz Nitro Express Reveg and Grafing Experiment.

    Quote Originally Posted by Ph1 View Post
    Interesting stuff pugz

    Can help a lot with people willing to try it out

    Maybe make a sticky for it in the advanced growing techniques section?
    Feel free mate, I'm using an IPad and takes along time due to 5 pic thing
    Will be far easier if using a pc or lappie.

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  13. #17

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    Quote Originally Posted by Pugz View Post
    Feel free mate, I'm using an IPad and takes along time due to 5 pic thing
    Will be far easier if using a pc or lappie.
    Same here mate, iPad aswell haha

    If the time comes ill pop it in there

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    Good luck with the experiment Pugz and thanks for posting all that info.

    All good things
    TDA

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    Default Pugz Nitro Express Reveg and Grafing Experiment.

    Quote Originally Posted by Ph1 View Post
    Same here mate, iPad aswell haha

    If the time comes ill pop it in there
    No problem mate.

    Would be kinda nice to have a sticky to my name

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    Default Pugz Nitro Express Reveg and Grafing Experiment.

    Quote Originally Posted by THE DUDE ABIDES View Post
    Good luck with the experiment Pugz and thanks for posting all that info.

    All good things
    TDA
    Hey dude nice to see ya.

    Thanks mate, I try my best with the iPad I but it takes so much messing to get things posted properly,
    Can't just copy and past the same as a pc.
    And the fucking curser, trying to edit a post is a fucking nightmare, I end up deleted half of the post just to amend a mistake

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