Agrobacterium rhizogenes transformation and expression of rol genes in kalanchoe

Patent metadata

Field	Value
Publication number	US-10329573-B2
Application number	US-201514875275-A
Country	US
Kind code	B2
Filing date	Oct 5, 2015
Priority date	Aug 17, 2012
Publication date	Jun 25, 2019
Grant date	Jun 25, 2019

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 method for reducing the height of a Kalanchoë interspecific hybrid plant by about 5% to 60%, compared to a wild-type control plant, comprising:
(a) co-cultivating wild-type Agrobacterium rhizogenes with a Kalanchoë interspecific hybrid plant, wherein said Agrobacterium 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 include at least rolB, and wherein said one or more rol genes have a nucleic acid sequence selected from the group consisting of SEQ ID Nos: 3, 4, 5 and 6, respectively and variants of SEQ ID NOs: 3, 4, 5, and 6 that encode a polypeptide selected from the group consisting of SEQ ID NOs: 19, 20, 21, and 22, respectively;
(b) selecting a putatively transformed root of said plant that comprises said one or more rol genes, which root has a hairy root phenotype;
(c) growing said root on a regeneration medium;
(d) regenerating a shoot from said root, thereby generating a plantlet,
(e) growing said plantlet into a mature plant, and
(f) selecting a plant having reduced height compared to a non-transformed control 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 interspecific hybrid Kalanchoë plant tissue with Agrobacterium rhizogenes , wherein said Agrobacterium 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 include at least rolB, and wherein said one or more rol genes have a nucleic acid sequence selected from the group consisting of SEQ ID Nos: 3, 4, 5 and 6, respectively and variants of SEQ ID NOs: 3, 4, 5, and 6 that encode a polypeptide selected from the group consisting of SEQ ID NOs: 19, 20, 21, and 22, 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;
(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 rol-transformed Kalanchoë interspecific hybrid has been transformed with one or more rol genes, 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 include at least rolB, and wherein said one or more rol genes have a nucleic acid sequence selected from the group consisting of SEQ ID Nos: 3, 4, 5 and 6, respectively and variants of SEQ ID NOs: 3, 4, 5, and 6 that encode a polypeptide selected from the group consisting of SEQ ID NOs: 19, 20, 21, and 22, respectively.

5. Progeny of a rol transformed Kalanchoë interspecific hybrid of claim 4 , wherein said progeny comprises at least one said rol gene integrated into its genome, and wherein said progeny is an interspecific hybrid that exhibits said intermediate height of the hybrid of claim 4 .

6. The method of claim 1 , wherein the nucleic acid sequence for rolA, rolB, rolC, and rolD variants has at least 98% sequence identity with SEQ ID NOs: 3, 4, 5, and 6, respectively.

7. The method of claim 1 , wherein the nucleic acid sequence for rolA, rolB, rolC, and rolD variants has at least 99% sequence identity with SEQ ID NOs: 3, 4, 5, and 6, respectively.

8. The method of claim 3 , wherein the nucleic acid sequence for rolA, rolB, rolC, and rolD variants has at least 98% sequence identity with SEQ ID NOs: 3, 4, 5, and 6, respectively.

9. The method of claim 3 , wherein the nucleic acid sequence for rolA, rolB, rolC, and rolD variants has at least 99% sequence identity with SEQ ID NOs: 3, 4, 5, and 6, respectively.

10. The rol-transformed Kalanchoë interspecific hybrid with intermediate height of claim 4 , wherein the nucleic acid sequence for rolA, rolB, rolC, and rolD variants has at least 98% sequence identity with SEQ ID NOs: 3, 4, 5, and 6, respectively.

11. The rol-transformed Kalanchoë interspecific hybrid with intermediate height of claim 4 , wherein the nucleic acid sequence for rolA, rolB, rolC, and rolD variants has at least 99% sequence identity with SEQ ID NOs: 3, 4, 5, and 6, respectively.

Assignees

• 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

• Y02A40/146

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

• C12N15/8205 Primary

  Agrobacterium mediated transformation · CPC title

• A01H3/00

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

• A01H1/00

  Processes for modifying genotypes {; Plants characterised by associated natural traits} (A01H4/00 takes precedence) · CPC title

• A01H5/02

  Flowers · CPC title