Multicellular living organisms and unmodified parts thereof and – Plant – seedling – plant seed – or plant part – per se – Higher plant – seedling – plant seed – or plant part
Reexamination Certificate
1999-09-27
2001-05-08
Benzion, Gary (Department: 1649)
Multicellular living organisms and unmodified parts thereof and
Plant, seedling, plant seed, or plant part, per se
Higher plant, seedling, plant seed, or plant part
C800S274000, C800S298000, C800S260000, C047S05810R
Reexamination Certificate
active
06229072
ABSTRACT:
FIELD OF THE INVENTION
This invention relates to improved plants. In particular, it relates to new plant germplasm of the Brassica species, having a reduced content of undesired glucosinolates.
BACKGROUND OF THE INVENTION
Economic production of Brassica spp. hybrids requires a pollination control system and effective transfer of pollen from one parent to the other. The ogura cytoplasmic male sterility (cms) system, developed via protoplast fusion between radish (
Raphanus sativus
) and rapeseed (
Brassica napus
) is one of the most promising methods of hybrid production. It provides stable expression of the male sterility trait (Ogura 1968), Pelletier et al. (1983) and an effective nuclear restorer gene (Heyn 1976).
Initial restorer material showed reduced female fertility which was overcome through backcrossing. Delourme et al. (1991) attributed this to elimination of a portion of the radish chromosome that had been introduced along with the restorer gene. In their work, successive backcross generations produced fertility levels successively closer to normal.
High glucosinolate (GSL) content in seed of
Brassica napus
is an anti-nutritional factor. Meal made from such seed is unsuitable for use in animal feeds. Seed GSL level is an expression of the genotype of the female plant and is determined by four to eight separate dominant and additive genes. Two to five genes are involved in alkenyl (one of the aliphatic group) glucosinolate content, while two or three genes are involved in indole glucosinolate content (Rücker and Röbbelen, 1994). Total aliphatics may be determined by up to six genes (Magrath et al. 1993).
SUMMARY OF THE INVENTION
An object of the present invention is to provide Brassica spp. hybrids, seeds, microspores, ovules, pollen, vegetative parts containing low glucosinolate and the restorer gene.
Yet another object of the present invention is to provide interspecific crosses using fertile, low glucosinolate plants with the ogura cytoplasm as the female, followed by selection for fertility and low glucosinolate.
A further object of the invention is to provide a method for identifying a restorer line that contains only the portion of the
Raphanus sativus
material necessary for fertility and not the portion of the
Raphanus sativus
material that produces high glucosinolate.
Accordingly, our invention comprises a gene restorer line of
Brassica napus
which contains a
Raphanus sativus
restorer gene but is essentially free of
Raphanus sativus
glucosinolate-producing genes. In particular, we provide the gene restorer line KH, and progeny derived therefrom, seed of which is low in glucosinolates. We further provide
Brassica napus
restorer lines free of glucosinolate-producing genes having a characteristic RFLP signature, as hereinafter described, and a method of producing such lines which comprises crossing
Brassica napus
restorer and/or hybrid lines with desired
Brassica napus
germplasm and selecting progeny having a characteristic RFLP signature. Clearly this invention encompasses hybrids containing the restorer gene without the high glucosinolate material. Additionally, these hybrids can be used to create new restorer lines within the scope of this invention.
The present invention broadly includes a method of producing an improved restorer line of Brassica for use in a cytoplasmic male sterility system, which comprises forming a plant population from a gene restorer line of
Brassica napus
which contains a
Raphanus sativus
restorer gene and
Raphanus sativus
glucosinolate genes. Then breeding with the progeny of the plant population. Furthermore, it includes testing the progeny for fertility indicating the
Raphanus sativus
restorer gene is present and for levels of glucosinolate wherein the presence and absence of
Raphanus sativus
high glucosinolate production is shown; and selecting progeny which are positive for presence of the restorer gene and negative for the
Raphanus sativus
with glucosinolate production.
The inventive methods of this application also include a method of forming
Brassica napus
hybrid seed and progeny thereof from a cytoplasmic male sterility system which includes a restorer line containing
Raphanus sativus
restorer gene. This method includes the steps of providing a homozygous improved restorer line produced, as outlined above, using the restorer line in a hybrid production field as the pollinator; using cytoplasmic male sterile plants in a hybrid production field as the hybrid seed producing plant; and harvesting the hybrid seed from the male sterile plant.
Additionally, when producing progeny, the method includes the step of planting the hybrid seed from the male sterile plant and growing a plant therefrom.
The present invention clearly shows how to form an improved Brassica ssp., an improved
Brassica napus
plant, having low glucosinolate seeds, the plant containing
Raphanus sativus
gene material that is capable of restoring fertility to the ogura cytoplasmic male sterile plants, the improvement comprising an improved
Brassica napus
plant evidencing deficient glucosinolate production from the
Raphanus sativus
material, wherein the improved plant produced low glucosinolate seeds.
A
Brassica napus
plant containing
Raphanus sativus
restorer gene unlinked from
Raphanus sativus
glucosinolate genes adapted to restore fertility to oguracytoplasmic male sterile.
The present invention describes the molecular marker method. This is a method wherein the markers mapping to similar regions as those in the group consisting of, WG3F7, TG1H12, OPC2, WG4D10, WG6F3 are employed to identify the
Raphanus sativus
material which contains high glucosinolate producing genes.
The present invention encompasses not only canola quality but any low glucosinolate material produced for a cytoplasmic sterile plant containing
Raphanus sativus.
Any canola quality (erucic acid<2% and <30 &mgr;moles glucosinolates/gram defatted dry meal) restorer line, capable of inducing fertility in Brassica plants containing the INRA Ogura cytoplasmic male sterility. Further, the present invention encompasses Brassica spp. hybrids, seeds, microspores, ovules, pollen, vegetative parts containing low glucosinolate restorer gene. Interspecific crosses using fertile, low glucosinolate plants with the ogura cytoplasm as the female, followed by selection for fertility and low glucosinolate.
Brassica spp. hybrids, seeds, microspores, ovules, pollen, vegetative parts containing low glucosinolate restorer gene as identified by using probes such as those as described herein.
Additionally in the broad scope of the invention included is the
Brassica napus
(spring and winter types) or
B. rapa
containing the low glucosinolate restorer gene as described.
REFERENCES:
patent: 4658084 (1987-04-01), Beversdorf et al.
patent: 5356799 (1994-10-01), Fabijanski et al.
patent: 5478369 (1995-12-01), Albertsen et al.
patent: 5973233 (1999-10-01), Burns et al.
patent: 652964 (1992-04-01), None
patent: 599042 (1994-06-01), None
patent: 671121 (1995-09-01), None
Ecke et al. Bericht uber die Arbeitstagung der Arbeitsgemeinschaft der Saatzuchtleiter im Rahmen der Vereingung Osterreichsher Pflanzanzuchter, vol. 44, pp. 75-84 (translation pp. 1-28), 1993.*
Mailer et al. Canadian Journal of Plant Science, vol. 70, pp. 399-407, 1990.*
Visentin et al. Journal of Agricultural Food Chemistry, vol. 40, pp. 1687-1691, 1992.*
R. Delourme, A. Bouchereau, N. Hubert, M. Renard, B.S. Landry. Identification of RAPD Markers Linked to a Fertility Restorer Gene for Ogura Radish Cytoplasmic Male Sterility of Rapeseed (Brassica Napus L.). Theoretical Applied Genetics, vol. 88, pp. 741-748, 1994.
R. Pellan-Delourme and M. Renard. Cytoplasmic male sterility in rapeseed (Brassica Napus L.): female fertility of restored rapeseed with “Ogura” and cybrids cytoplasms. Genome, vol. 30, 1988.
F.W. Heyn. Transfer of Restorer Genes from Raphanus to Cytoplasmic Male Sterile Brassica napus and the Generics of Fertility Restoration. Crucierase Newsletter 1:15-16, 1976.
R. Delourme, R. and F. Eb
Barnes Steve
Burns Dale R.
Buzza Greg C.
Forhan Mark A.
Huskowska Teresa
Adventa Technology Ltd
Benzion Gary
Kimball Melissa L.
Rewoldt Dana
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