Plant Breeding Breakthrough - Offspring with Genes From Only One Parent

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I wonder what this could mean for the future of cannabis breeding.


Plant Breeding Breakthrough: Offspring With Genes from Only One Parent

ScienceDaily (Mar. 25, 2010) — A reliable method for producing plants that carry genetic material from only one of their parents has been discovered by plant biologists at UC Davis. The technique, to be published March 25 in the journal Nature, could dramatically speed up the breeding of crop plants for desirable traits.

The discovery came out of a chance observation in the lab that could easily have been written off as an error.


"We were doing completely 'blue skies' research, and we discovered something that is immediately useful," said Simon Chan, assistant professor of plant biology at UC Davis and co-author on the paper.


Like most organisms that reproduce through sex, plants have paired chromosomes, with each parent contributing one chromosome to each pair. Plants and animals with paired chromosomes are called diploid. Their eggs and sperm are haploid, containing only one chromosome from each pair.
Plant breeders want to produce plants that are homozygous -- that carry the same trait on both chromosomes. When such plants are bred, they will pass the trait, such as pest resistance, fruit flavor or drought tolerance, to all of their offspring. But to achieve this, plants usually have to be inbred for several generations to make a plant that will "breed true."


The idea of making a haploid plant with chromosomes from only one parent has been around for decades, Chan said. Haploid plants are immediately homozygous, because they contain only one version of every gene. This produces true-breeding lines instantly, cutting out generations of inbreeding.


Existing techniques to make haploid plants are complicated, require expensive tissue culture and finicky growing conditions for different varieties, and only work with some crop species or varieties. The new method discovered by Chan and postdoctoral scholar Ravi Maruthachalam should work in any plant and does not require tissue culture.


Ravi and Chan were studying a protein called CENH3 in the laboratory plant Arabidopsis thaliana. CENH3 belongs to a group of proteins called histones, which package DNA into chromosomes. Among the histones, CENH3 is found only in the centromere, the part of the chromosome that controls how it is passed to the next generation.


When cells divide, microscopic fibers spread from each end of the cell and attach at the centromeres, then pull the chromosomes apart into new cells. That makes CENH3 essential for life.


Ravi had prepared a modified version of CENH3 tagged with a fluorescent protein, and was trying to breed the genetically modified plants with regular Arabidopsis. According to theory, the cross should have produced offspring containing one mutant gene (from the mother) and one normal gene (from the father). Instead, he got only plants with the normal gene.
"At first we threw them away," Chan said. Then it happened again.
Ravi, who has a master's degree in plant breeding, looked at the plants again and realized that the offspring had only five chromosomes instead of 10, and all from the same parent.


The plants appear to have gone through a process called genome elimination, Chan said. When plants from two different but related species are bred, chromosomes from one of the parents are sometimes eliminated.
Genome elimination is already used to make haploid plants in a few species such as maize and barley. But the new method should be much more widely applicable, Ravi said, because unlike the process for maize and barley, its molecular basis is firmly understood.


"We should be able to create haploid-inducing lines in any crop plant," Ravi said. Once the haploid-inducing lines are created, the technique is easy to use and requires no tissue culture -- breeders could start with seeds. The method would also be useful for scientists trying to study genes in plants, by making it faster to breed genetically pure lines.
After eliminating half the chromosomes, Chan and Ravi had to stimulate the plants to double their remaining chromosomes so that they would have the correct diploid number. Plants with the haploid number of chromosomes are sterile.


The research also casts some interesting light on how species form in plants. CENH3 plays the same crucial role in cell division in all plants and animals. Usually, such important genes are highly conserved -- their DNA is very similar from yeast to whales. But instead, CENH3 is among the fastest-evolving sequences in the genome.


"It may be that centromere differences create barriers to breeding between species," Chan said. Ravi and Chan plan to test this idea by crossing closely-related species.


The work was supported by a grant from the Hellman Family Foundation.
 
I'd think Selfing the plants would do the same thing since you only want genes from one parent. When you self your only using 1 parent so there is only 1 parents genes....but I'll admit I only scanned the article I'm in a hurry lol.
 
Great read but....how do you do it? I may just be high but I didn't get any useful information out of that article on how to do this in my garden. Also, can you really get a Masters in plant breeding? I would think you'd call it a Masters in Horticulture but maybe the writer of the article changed the words around in an attempt to be easily readable to the uninformed.
 
I have that paper. Here is a cut and paste from a post i made on another site...


I haven't read it all yet. I just went for the meat Here is what I understand they did.
Soak some wild type seeds in ethylmethane sulphonate, grow them out and take clones, The plants grown out get tested to see if they mutated and the mutated DNA will produce cenh3-1. Some plants will and some wont mutate properly so testing is needed. Using the identified, mutated clones, take tons of pollen from many males and flood the pistil with it (very few mutated pollen grains are viable) to get a good seed set they used 60 anthers per pistil. Some of these seeds will be haploid, others diploid some cases Aneuploids, so testing is done again. When the diploid plants are selfed they form an instant IBL.

end of cut and paste

This will work for hybrids as well, although there will be huge variability in the Mutant X Non Mutant seed lot, if done proporly the next generation is the IBL same as wild types. Oh yeah one more thing, they used Monoecious plants in the experiment.

Da'Shadow
 
:thumbsup:wrappin' it up as usual DaShadow.....thanks for the summary.....


ganj on.....
 
Hey Da Shadow, he all,

Can you please post the entire paper.

And Yes, I can verify that a Masters in Plant Breeding degree does exist, the offer it at my university, as well as the undergrad plant breeding degree and a PhD.

EMS (ethylmethane sulfonate) is totally random in its insertion into DNA, and you have to screen many thousands of plants in its tradtional use, thats why I want to see the whole paper. If there isn't a more reliable screening method, then I think the method of double haploid production sounds easier and faster, and TC isn't all that hard to do if you can read up on it and have a little lab experience.

Be safe

A
 
I caught these guys talking about this on my local NPR station. I was screamin at the radio...CANNABIS-use it on CANNABIS!!! Then at the end, the interviewer says 'I hear the cannabis growers are interested'-"Yes, I did an internet search on my research paper and it brought up 7 diff pot sites in 3 dif languages.'

So if it works it cuts down the breeding generations to make IBL's
They said it was applicable to any species
I'm still researching it


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Haploid Plants through Seeds

Tech ID: 19877 / UC Case 2010-030-0

Abstract

Researchers at the University of California Davis have developed a novel method to produce haploid plants through seeds. This method induces genome elimination (from one parent in a cross) with a precise mutation, rather than by culturing haploid cells or by crossing distantly related plants.

Full Description

Plant breeding relies on screening numerous plants to identify novel, desirable characteristics. Very large numbers of progeny from crosses often must be grown and evaluated over several years in order to select one or a few plants with a desired combination of traits.

Standard breeding of diploid plants often requires screening and back-crossing of a large number of plants to achieve the desired genotype. One solution to the problem of screening large numbers of progeny has been to produce haploid plants, the chromosomes of which can be doubled using colchicine or other means to achieve instantly homozygous, doubled-haploid plants.

With doubled haploid production systems, homozygosity is achieved in one generation. Thus, the breeder can eliminate the numerous cycles of inbreeding necessary to achieve practical levels of homzygosity using conventional methods. Indeed, true homozygosity for all traits is not achievable by conventional breeding methods.

Existing methods of generating haploid plants have numerous disadvantages. Culturing of haploid cells is expensive and laborious, and some species have proven recalcitrant to this technique. Crossing to a distantly related species (wide crosses) causes genome elimination in only a small number of species, and almost always requires embryo rescue in vitro to generate viable plants. Haploid-inducing lines in maize are genetically complex and yield haploids at low efficiency. All current methods may be extremely dependent on genotype. UC Davis researchers have developed a method of inducing haploids in a cross between plants of the same genotype which is based on exploitation of a universal feature of eukaryote chromosomes and which yields haploid plants from seeds.



Applications

This novel technology has multiple applications, including:

* Doubled haploid plants can rapidly create homozygous F2s from a hybrid F1.
* Haploid plants are very useful for genomics because they contain only one version of each gene.
* The method can transfer paternal chromosomes into maternal cytoplasm. Thus, it can create cytoplasmic male sterile lines with a desired genotype in a single step. Currently, generating a cytoplasmic male sterile line with a desirable genotype requires many generations of backcrossing.

Advantages

Using this innovative method:

* Genome elimination can be engineered with a precise molecular change that is not dependent on parental genotype. The gene that is manipulated is found in all eukaryotes and serves a universal function. Thus, haploid plants can be made in species where conventional methods, such as tissue culture of haploid cells and wide crosses, are typically unsuccessful.
* No tissue culture is required. Haploids are produced through seed by simple genetic crosses. This will greatly reduce the cost and labor required for haploid plant production, and make the process accessible to breeders lacking specialized expertise in culturing haploid cells.
* Plants from exactly the same cultivar can be crossed to eliminate one parental genome using a precise genetic change. This greatly simplifies synchronizing flowering time and readiness to cross (relative to the wide cross method of haploid production).
* This method yields haploid plants much more efficiently than current wide crossing protocols, or existing haploid inducers in maize.
* Apart from haploid-inducing lines in maize, this is the only known method of producing haploid plants in which paternal chromosomes are transferred into maternal cytoplasm, generating cytoplasmic male sterile lines with a desired genotype in a single step.

Related Materials

* Ravi M, Chan SW. 2010. Haploid plants produced by centromere-mediated genome elimination. Nature. 464(7288):615-8. - 03/25/2010

PATENT STATUS

* Patent Pending

INVENTORS

* Chan, Simon R.
* Maruthachalam, Ravi

Other Information
Categorized As

* Agriculture & Animal Science
o Plant Traits
o Transgenics
* Biotechnology
o Food
o Industrial/ Energy
* Research Tools
o Other

Related cases

2010-030-0
Keywords

haploid, plant, cultivar, breeding, transgenic
Contact

Randi L. Jenkins/ [email protected] / tel: 530-754-7650. Please reference Tech ID #19877.

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© 2009 - 2010, The Regents of the University of California

Link: http://techtransfer.universityofcalifornia.edu/NCD/19877.html
 
METHODS
Plant materials. cenh3-1 was isolated by the TILLING procedure in collaboration
with V. Sundaresan and L. Comai at the University of California, Davis31.
The TILLING population was created by mutagenizing Arabidopsis thaliana in
the Col-0 accession with ethylmethane sulphonate, using standard protocols.We
isolated cenh3-1 by TILLING using the CEL1 heteroduplex cleavage assay, with
PCR primers specific for the CENH3/HTR12 gene.
cenh3-1 is predicted to disrupt normal splicing ofCENH3, because it mutates a
conserved splice acceptor site at the beginning of the second coding exon.
Translation of an mRNA containing the first coding exon spliced to an incorrect
location within CENH3 is predicted to yield only 18 correct amino acids. As the
histone-fold domain of CENH3 begins at amino acid residue 82, we believe that
cenh3-1 is a null allele (this is supported by its embryo-lethal phenotype).
Cloning of the GFP–CENH3 and GFP–tailswap transgenes, and construction
of the complemented cenh3-1 GFP–CENH3 and cenh3-1 GFP–tailswap lines, are
described elsewhere (M.R. et al., manuscript in preparation). Primer sequences
and full details are available on request.
To cross wild type as the female to GFP–tailswap as the male, we used a
dissecting microscope to observe directly pollen deposition on the stigma
(GFP–tailswap is mostly male sterile). The amount of viable pollen in individual
flowers of GFP–tailswap varies. We selected flowers that clearly showed
higher amounts of pollen, and pollinated with more than 60 anthers (10 GFP–
tailswap flowers) per wild-type stigma to achieve the seed set reported in
Table 1. Using an optivisor (magnifying lens) and approximately 12 anthers
(2 GFP–tailswap flowers) per wild-type stigma, we obtained a much lower seed
set per siliqua.
The percentage of normal seeds was determined by visual inspection using a
dissecting microscope. Seeds from GFP–tailswap3wild-type crosses were sown
on 13 MS plates containing 1% sucrose to maximize germination efficiency,
particularly of seed that had an abnormal appearance. Late germinating seeds
were frequently haploid.
The quartet mutant that we used was qrt1-2 (ref. 32). Male sterility in the C24/
Ler line was conferred by the A9-barnase transgene33,34.
In the GFP–tailswap 3 Wa-1 experiment, progeny from the GFP–tailswap 3
Wa-1 cross that contained only Wa-1 chromosomes were confirmed as diploid
using chromosome spreads. We scored plants that were heterozygous for some
chromosomes (Col-0 and Wa-1 markers) and homozygous for other chromosomes
(Wa-1 markers only) as aneuploid. We did not find triploid offspring
(heterozygous for markers on all chromosomes). A subset of plants was further
karyotyped by means of chromosome spreads to confirm aneuploidy.
Cytogenetic analysis. Mitotic and meiotic chromosome spreads from anthers
were prepared according to published protocols35.
Colchicine treatment. Colchicine treatment of developing haploid plants used a
previously published protocol with minor modifications36. A solution of 0.25%
colchicine, 0.2% Silwet was prepared, and a 20 ml drop was placed on the meristem
before bolting. Plants became transiently sick after colcichine treatment.
Upon recovery, fertile inflorescences appeared from secondary meristems indicating
successful chromosome doubling. Haploid plants can also be treated after
bolting, although the rate of success is considerably lowe
 
Now, a question from the sativa-smoking contingent..

Yikes! What about creating haploid Cannabis males!

Could this be the literal 'killer botanical app' for cannabis eradication?

Would these males throw pollen that would fertlize the girls, but result in sterile seeds, so the next generation of beans don't germinate? No grandkids? Will the UN adopt this to help eradicate field-grown pot in 3rd world countries.

Paranoid minds want to know.
 
I'm an "old fashioned" breeder and this info does sound like a breakthrough, though as described, beyond the reach of any common grower/ breeders in technique and equip.!

I didn't take this the way Clancy D did...though the male sterility factor did alarm me some.

The advantages of quickly producing IBL'S is astounding but the patenting issues and the the application ranges of the patent application ('s), is a bit unnerving also.

I can't believe, or don't wish to believe that a very few MAD scientists/ polititians, bankers etc. COULD use such science, for crop eradication of such a vast, asset as one of the most valuable plants on the planet and be able to get away with it. Let's also think of other crops! This kind of thing IS happening worldwide but seed savers are modern saints!

If "they" are smart they could profit from the herb and lots of diversity. Probably "they" want to have only "THEM" manipulate the future! MAD EGOS! Not likely to happen long term on this planet or in a diverse universe. Only my experience and thinking. ( my opinion )

Perhaps "they" are completely MAD/crazy and think?...that all can be done through power, synthetic/ ARTIFICIAL means! Anyone studied in planet/ empire history can see through it!

I THINK a close watch should be kept on every level and by multiple " watchdog" groups for any misuse of science/ power, including our few watchdog scientist groups! Ultimately, our great mother earth,...god, goddess, universe, will balance things and cull bad things!

Very interesting topic though, I hope this thread can be "stickied" by admin for our viewing, comment, and any new developments. Please fellow Islanders, let's keep an eye out!

I hope that ALL outdoor sinse crops arn't endangered and that the only way to keep or preserve good and old canna and other "heirloom genetics" will not need to be done indoors with hepafilters and freezers! LOL Very unlikely and impractical, even to "them".

This would be nearly impossible even for the richest most controlling powermongers on earth and would cause large genotype/ phenotype plant drift over a short time.

What if there is an extended power outage? a planetary polar flip, even short term, will ruin all the artificial, electronic, information, money, banking, politics, petrol, transport, etc. in all countries, sectors of human affairs! Farmers with good seed will survive as always.

As always, long term, even eternally, CHANGE IS THE ONLY CONSTANT! LOL!

I prefer to stay positive AND watchful. My best to you and yours. Peace, Brwin.
 
Gojo, I'm an old fasioned grower and breeder, the info you posted though is way out there, and a bit techie, even for me. I'd like to see other breeders take on this info. ...man.

This emphasis on PLOYDY seems over the top to me. Can you quantify and or qualify your deductions, that our canna crops of carefully selected breeding are in danger? FACTS, possible impacts, in different secluded areas, etc. Somehow I smell a rat. ( negative )

Kinda sounds like gvt. manipulation of minds to make growers feel powerless, not like this is a new ploy or tactic, they've been at for like 70 yrs. methodically. Peace, Brwin.
 
Great thread and comments!!! Mostly over my head, but I get the bottom lines. Great Sticky. Thanks to MJ and all who posted. Subscribed. :thumbsup:
 
Hey Gojo;

I have been following a few of your threads, thanks for contributing :) lots of information that is spot on (imo) I think you offered to get scientific papers for peeps if they neded them :) i was wondering if you may have (or access to)

Genetic and Hormonal Regulation of Growth, Flowering, and Sex Expression in Plants, by M. Kh. Chailakhyan © 1979 Botanical Society of America.

I would love to read this paper... before asking you a few questions..

Again thatkx for the contributions

Da'Shadow
 
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