tissue culture

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mnemko
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tissue culture

Post: # 69723Post mnemko
Tue Apr 16, 2019 8:15 pm

When one has a seedling he'd like to have tested in multiple locations, does one make cuttings (e.g., 1-node, 3-node) or is tissue culture a viable option? If so, is there some lab that does that?
Marty Nemko
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Larry Davis
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Re: tissue culture

Post: # 69733Post Larry Davis
Fri Apr 19, 2019 9:29 am

So far as I'm aware there has been relatively little interest in propagation by tissue culture, at least acknowledged as such, for several decades. I looked into it with a couple undergraduates when there was someone at U of Wisconsin hort dept doing it. And one of the large companies J & P I think, promoted at least one yellow floribunda that was propagated that way. For me it was a dog and not hardy. The whole idea faded away after a couple years. The Japanese blue rose of course (Suntory Co.) came through some stages of culture. But my impression is that the Dutch found single leaf cuttings or stentlings (grafted) were the most efficient way to bulk up a new line. I've use single leaf cuttings with a number of CV. I've also found that a good CV under lights can increase more than 20-fold in a year if you pay attention and feed well with regular cuttings of 3-4 nodes. When a cutting grows a new shoot cut it and propagate. You can easily get new ones every 8-10 weeks, allowing around 2 to the 5th power plants in a year. For CV selected for own-root growth, like the minis from Nor-East were, really simple. That's my experience.

Don
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Re: tissue culture

Post: # 69738Post Don
Sun Apr 21, 2019 12:08 am

Armstrong was using it back in the last millennium as was High Country though I couldn't get much detail from High Country when I was looking into it. I think Dobros dabbled in it for Conard Pyle/Star for a bit. Weston Nurseries was using it for woodies back then too but they didn't do much in the way of roses. There are some protocols that work well but I doubt anyone is currently applying them to commercial production. One of the dark secrets of tissue culture is that many times the resulting plants are phenotypically inconsistent and I know this was the case for roses.

Ralph Moore had a bud propagation protocol that was highly productive in the manner described by Larry for leaf cuttings, basically a refinement where he chose a particular bit of budwood at the leaf base for propagation. I think Kim can tell you more about it and there is a description by Ralph himself in either text or video or both iirc.

Commercial producers have gone contract now. Conard Pyle was among the last to do their own propagation but switched over to contracting with Greenhart which may be your best bet if you get to wanting bigger numbers.
What doesn't kill them makes them stronger.

roseseek
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Re: tissue culture

Post: # 69739Post roseseek
Sun Apr 21, 2019 2:24 am

Ralph simply used one bud cuttings under mist. What he showed me wasn't anything special. He just cut up longer cuttings into pieces containing a bud near the top and struck them into perlite under mist. As usual, pretty much everything he stuck in that house rooted.
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Don
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Re: tissue culture

Post: # 69740Post Don
Sun Apr 21, 2019 7:00 am

I recall he claimed that roots were most likely to sprout from the tissue opposite a pair of bud eyes. Cuttings with that part of the stem exposed worked best. I'm trying to find the notes I have on it.
What doesn't kill them makes them stronger.

Karl K
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Re: tissue culture

Post: # 69746Post Karl K
Mon Apr 22, 2019 8:29 pm

Some plants can be propagated by leaf cuttings, as most gardeners know. It hadn't occurred to me that roses might be among them.

Parsons (1847)
Some years since, Lecoq, a French cultivator, conceived the idea of endeavoring to propagate roses by the leaf. He gathered some very young leaves of the Bengal rose, about one quarter developed, cutting them off at their insertion, or at the surface of the bark. He planted these in peat soil, in one‑inch pots, and then plunged the pots into a moderate heat. A double cover of bell glasses was then placed over them, to exclude the air entirely, which course of treatment was pursued until they had taken root. The shortest time in which this could be accomplished was eight weeks, and the roots were formed in the following manner. First, a callus was formed at the base of the leaf, from which small fibres put forth; a small bud then appeared on the upper side (figure 16); a stalk then arose from this bud, which finally expanded into leaves and formed a perfect plant
http://bulbnrose.x10.mx/Roses/breeding/ ... f1847.html

This sounded promising ... until I found that Lecoq's results were not spectacular. In fact, few of the leaves produced plants. Even so, subsequent research does indicate that various tissues might be cultured. Lloyd et al., 1988 reported:
Shoots with up to 3 simple leaves were formed on excised leaves of R. persica X xanthina, R. laevigata and R. wichuraiana only (Table 1). They appeared within six weeks of inoculation onto medium containing 0.5 mg l-1 BAP and no NAA (Table 1). They were formed directly on the petiole and the midribs of leaflets without intervening callus. On transfer to multiplication medium, only shoots of R. persica X xanthina survived.
http://bulbnrose.x10.mx/Roses/breeding/ ... s1988.html

Other species and cultivars did not produce callus from leaves or roots.

Don
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Re: tissue culture

Post: # 69747Post Don
Mon Apr 22, 2019 9:13 pm

Callus can be induced by modifying the proportions of IBA, GA and kinetin in the culture medium. Roots or shoots can then be induced from the callus by changing the ratios again. My experience is with nicotiana but rosa protocols exist. I'll dredge up what I have and post them at some point, been busy.
What doesn't kill them makes them stronger.

Karl K
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Re: tissue culture

Post: # 69749Post Karl K
Tue Apr 23, 2019 11:46 am

Before I forget again, Kane (2000) wrote about micropropagation of roses. One comment is particularly interesting (I think).
"Many miniature rose cultivars display vigorous growth when established in vitro; the actual response is highly cultivar dependent."
http://bulbnrose.x10.mx/Roses/breeding/ ... e2000.html

This suggests that it should be possible to breed for this particular trait. Identify a collection of Minis that grow vigorously in vitro, then cross among them.

Whether such cultivars would be useful in practice is another matter. It's just that I am fascinated by obscure (cryptic) traits that are subject to selection.

Oh, and Kane also mentioned: " In vitro-produced flowers are smaller and have fewer petals than these produced on plants grown in the greenhouse." These might be useful for breeding cultivars that normally produce too many petals and not enough pollen.

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