Plant transformation
AssigneeInari Agriculture Technology, Inc.
InventorJennifer A. Raji
Life of this patent
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Abstract
Methods and systems for transforming soybean and other leguminous plants in a hypertonic media without a transition through a callus stage are provided. In certain aspects, hypertonic media used in the methods and systems do not contain a growth hormone and/or a selection agent. In certain aspects, the transformation methods and systems provide a transformation efficiency of at least about 30%.
Claims19
A method of providing a transformed soybean plant, the method comprising the steps of:
- (a) providing a soybean plant quarter-seed meristem explant comprising the whole embryo of a source soybean seed, wherein the upper half of one cotyledon is left attached to the whole embryo and one cotyledon is completely removed, wherein the shoot apical meristem tissue of the whole embryo is intact and is exposed by removal of all primary leaves, and wherein the radicle tip is trimmed;
- (b) contacting the quarter-seed meristem explant with a hypertonic medium;
- (c) contacting the explant with a suspension of Agrobacterium comprising a transformation construct;
- (d) co-cultivating the explant 4-5 days;
- (e) partially embedding the trimmed radicle end of the explant in shoot induction medium with the meristem side facing up and with the hypocotyl positioned upright;
- (f) incubating with biweekly changes to fresh shoot induction medium;
- and
- (g) transferring to shoot elongation medium;
- whereby a rooted soybean plantlet that comprises transformed germline cells having at least one genetic modification, in comparison to the unmodified genome of the embryo of the source soybean seed, effected by the transformation construct is regenerated within 8 weeks from the time of inoculation, without transition through a callus phase and at a transformation efficiency of at least 30%.
A system for Agrobacterium mediated plant transformation comprising (a) a soybean plant quarter-seed meristem explant that has been pre-treated with a hypertonic medium that does not comprise a growth hormone or a selection agent, wherein the quarter-seed meristem explant comprises the whole embryo of a source soybean seed, wherein the upper half of one cotyledon is left attached to the whole embryo and one cotyledon is completely removed, wherein the shoot apical meristem tissue of the whole embryo is intact and is exposed by removal of all primary leaves, and wherein the radicle tip is trimmed;
- and (b) an Agrobacterium comprising a transformation construct, whereby the system leads to improved transformation efficiency such that a rooted soybean plantlet that comprises transformed germline cells having at least one genetic modification, in comparison to the unmodified genome of the embryo of the source soybean seed, effected by the transformation construct, is regenerated within 8 weeks from the time of inoculation, without transition through a callus phase and at a transformation efficiency of at least 30%.
Dependencies are inferred from the claim text.