Agrobacterium rhizogenes transformation and expression of rol genes in Kalanchoë

Patent metadata

Field	Value
Publication number	US-9253952-B2
Application number	US-201213588668-A
Country	US
Kind code	B2
Filing date	Aug 17, 2012
Priority date	Aug 17, 2012
Publication date	Feb 9, 2016
Grant date	Feb 9, 2016

Abstract

The present disclosure embraces Kalanchoë interspecific hybrid plants, and considers rol transformation in Kalanchoë species and hybrids. Disclosed herein are methodology and the like for producing rol-transformed Kalanchoë interspecific hybrid plants, as well as resultant rol-transformed Kalanchoë interspecific hybrid plants with novel phenotypes.

First claim

What is claimed is:

1. A species-independent method for transforming a Kalanchoë interspecific hybrid plant, comprising:
(a) co-cultivating wild-type A. rhizogenes with a Kalanchoë interspecific hybrid plant, wherein A. rhizogenes transfers one or more rol genes into said plant, wherein said one or more rol genes are selected from the group consisting of rolA, rolB, rolC, and rolD, wherein said one or more rol genes at least includes rolB, wherein the nucleic acid sequence for rolA, rolB, rolC, and rolD has 100% sequence identity with SEQ ID NOs: 3, 4, 5, and 6, respectively;
(b) selecting a putatively transformed root having a hairy root phenotype;
(c) growing said root on a regeneration medium;
(d) regenerating a shoot from said root, thereby generating a plantlet, and;
(e) growing said plantlet into a mature plant.

2. The method of claim 1 , further comprising assaying the presence of one or more rol genes in said mature plant.

3. A method for reducing the height of a Kalanchoë interspecific hybrid plant by about 5% to about 60%, compared to a wild-type control plant, comprising:
(a) transforming Kalanchoë plant tissue with A. rhizogenes , wherein A. rhizogenes delivers and integrates one or more rol genes into hybrid plant genome, wherein said one or more rol genes are selected from the group consisting of rolA, rolB, rolC, and rolD, wherein said one or more rol genes at least includes rolB, wherein the nucleic acid sequence for rolA, rolB, rolC, and rolD has 100% sequence identity with SEQ ID NOs: 3, 4, 5, and 6, respectively;
(b) selecting a putatively transformed root having a hairy root phenotype;
(c) growing said root on a regeneration medium;
(d) regenerating a shoot from said root, thereby generating a plantlet; and
(e) growing said plantlet into a mature plant, and;
(f) selecting a plant having a reduced height by about 5% to about 60% compared to the height of a non-transformed control plant of the same species.

4. A rol-transformed Kalanchoë interspecific hybrid with intermediate height, wherein said intermediate height is a reduction of height by about 5% to about 60% of the height of a control, non-transformed Kalanchoë interspecific hybrid plant, wherein the rot-transformed Kalanchoë interspecific hybrid has been transformed with one or more rol genes are selected from the group consisting of rolA, rolB, rolC, and rolD, wherein said one or more rol genes at least includes rolB, wherein the nucleic acid sequence for rolA, rolB, rolC, and rolD has 100% sequence identity with SEQ ID NOs: 3, 4, 5, and 6, respectively.

5. Progeny of a rol transformed Kalanchoë interspecific hybrid of claim 4 .

Assignees

• Christensen Ellen Margrethe Skovsgaard
• Müller Renate Petra Brigitte
• Lütken Henrik Vlk
• Hegelund Josefine Nymark
• Jensen Line
• Christensen Brian
• Jensen Else Bollerup
• Nielsen Kai Lønne
• Univ Copenhagen
• Knud Jepsen As

Inventors

• Christensen Ellen Margrethe Skovsgaard
• Müller Renate Petra Brigitte
• Lütken Henrik Vlk
• Hegelund Josefine Nymark
• Jensen Line
• Christensen Brian
• Jensen Else Bollerup
• Nielsen Kai Lønne

Classifications

• C12N15/8205 Primary

  Agrobacterium mediated transformation · CPC title

• C12N15/8261

  with agronomic (input) traits, e.g. crop yield · CPC title

• A01H3/00 Primary

  Processes for modifying phenotypes {, e.g. symbiosis with bacteria} (A01H4/00 takes precedence) · CPC title

• A01H5/0266

  Human Necessities · mapped topic

• Y02A40/146

  Genetically Modified [GMO] plants, e.g. transgenic plants · CPC title