Acacia

Summary

The genus Acacia is composed of as many as 8000 species native to the tropics and subtropics. The commercially valuable species Acacia mangium is variously known as “Mangium”, “Black Wattle” and “Hickory Wattle”, and is traded as brown salwood. Its distribution covers the northern part of Queensland in Australia, Papua New Guinea and the Molucca Islands of Indonesia. Mangium is a fast-growing tree, relatively short-lived (30-50 years) and adapted to a wide range of acidic soils in most tropical lowlands. It is a pioneer species that colonize disturbed sites. Major plantations of A. mangium are grown in Southeast Asia for paper pulp. Wood from this species is used in furniture, handles of sporting goods, door frames and light construction. Plantations of A. mangium are also used for erosion control, shade and shelter. This species is playing an important role in efforts to sustain commercial supply of tree products while reducing pressure on natural forest ecosystems.

The Institute of Molecular Agrobiology, located at the National University of Singapore, has filed applications related to the genetic transformation of A. mangium with Agrobacteriumand to the regeneration of the species. The PCT applications disclose methods for transforming A. mangium with a gene of interest using A. tumefaciens. One of the recited methods uses stems as initial tissue to be transformed with the bacterium.

For patent documents in the United States, Australia and New Zealand, the ownership has changed to Temasek Life Sciences Laboratory Ltd.

Specific Patent Information

Patent Number Title, Independent Claims and Summary of Claims Assignee
US 6846971

  • Earliest priority – 19 January 2000
  • Filed – 2 January 2002
  • Granted – 25 January 2005
  • Expected expiry – 18 January 2000
Title – Regeneration and genetic transformation of Acacia mangium

Claim 1
A method of transforming Acacia mangium with a gene of interest comprising the steps of:

a) activating Agrobacterium tumefaciens comprising said gene of interest by culturing said Agrobacterium in induction medium comprising acetosyringone;
b) preculturing an explant of Acacia mangium selected from the group comprising a stem, a leaflet, a petiole and a bud in medium comprising supplemented basic MS medium, wherein said explant is soaked in 0.5 M mannitol prior to the co-cultivation;
c) co-cultivating said activated Agrobacterium tumefaciens and said precultured explant in medium comprising supplemented basic MS medium to produce infected explants;
d) culturing said infected explants in medium comprising supplemented basic MS to induce callus and adventitious buds; and
e) culturing said callus or adventitious buds on a selective medium comprising supplemented basic MS medium; wherein in said supplemented basic MS medium comprises a) thidiazuron, b) indole-3-acetic acid, c) casein enzymatic hydrolysate, d) L-ascorbic acid, e) L-glutamine, f) L-asparagine, g) L-proline, h) sucrose and l) phytagel or agar.

Claim 7
A method of preparing transgenic Acacia mangium cells comprising the steps of
a) preculturing stem pieces of Acacia mangium in culture medium comprising supplemented basic MS medium, wherein said stem pieces are soaked in 0.5M mannitol prior to co-cultivating with Agrobacterium tumefaciens wherein said supplemented basic MS medium comprises a) thidiazuron, b) indole-3-acetic acid, c) casein enzymatic hydrolysate, d) L-ascorbic acid, e) L-glutamine, f) L-asparagine, g) L-proline, h) sucrose and i) phytagel or agar; and
b) co-cultivating said stem pieces of step (a) with Agrobacterium tumefaciens in culture medium AM-265, wherein said Agrobacteriumwas activated by culturing the Agrobacterium in induction medium comprising acetosyringone prior to said co-cultivation.

Granted US 6846971 recites a method of Agrobacterium-mediated transformation of Acacia mangium.

Temasek Life Sciences Laboratory Ltd

AU 784180 B2

  • Earliest priority – 19 January 2000
  • Filed – 19 January 2000
  • Granted – 12 February 2006
  • Expected expiry – 18 January 2020
Title – Regeneration and genetic transformation of Acacia mangium

Caim 1
A method for regenerating Acacia mangium comprising:
a) inducing callus formation from an explant;
b) culturing said callus to produce adventitious buds;
c) culturing said adventitious buds to elongate and produce pinnate leaves; and
d) culturing elongated buds of step (c) such that they produce roots and become plantlets.
Claim 11
A method for regenerating Acacia mangium comprising:
a) culturing auxiliary buds from an Acacia mangium tree to produce adventitious buds comprising hyllodes;
b) subculturing said adventitious buds comprising phyllodes to produce advantitious shoots;
c) culturing said adventitious shoots.
Claim 14
A method of tranforming Acacia mangium with a gene of interest comprising the steps of:
a) activating Agrobacterium tumefaciens comprising said gene of interest to form activated Agrobacterium tumefaciens;
b) preculturing an explant of Acacia mangium to yield a precultured explant;
c) co-cultivating said activated Agrobacterium tumefaciens and said precultured explant to produce infected explants;
d) culturing said infected explants to induce callus and adventitious buds; and
e) culturing said callus or adventitious buds on a selective medium.
Claim 25
A method for promoting root formation from transformed adventitious buds comprising

  • culturing transformed adventitious buds on a medium comprising 1/2 MS basic medium supplemented with a) alpha-naphthaleneacetic acid, b) kinetin, c) casein enzymatic hydrolysate, d) L-ascorbic acid, e) L-glutamine, f) L-asparagine, g) L-proline, h) sucrose and i) phytagel.
Claim 28
A method of preparing transgenic Acacia mangium cells comprising the steps of
a) preculturing stem pieces of Acacia mangium in a culture medium; and
b) cocultivating said stem pieces of step (a) with activated Agrobacterium tumefaciens.
Claim 41
A transgenic Acacia mangium cell.
Claim 42
A transgenic Acacia mangium plant.

The applicant of granted AU 784180 is stated in the patent document as Temasek Life Sciences Laboratory.

US 2005/155116 A1

  • Earliest priority – 19 January 2000
  • Filed – 9 December 2004
  • Application pending
Title – Regeneration and genetic transformation of acacia mangium

Claim 1
A method for regenerating Acacia mangium comprising:
a) inducing callus formation from an explant;
b) culturing the callus on a medium comprising thidiazuron to produce adventitious buds;
c) culturing the adventitious buds to elongate the buds and produce pinnate leaves; and
d) culturing elongated buds of step (c) to produce plantlets with roots.
Claim 11
A method for regenerating Acacia mangium comprising:
a) culturing auxiliary buds from an Acacia mangium tree to produce adventitious buds comprising phyllodes;
b) subculturing the adventitious buds comprising phyllodes to produce adventitious shoots;
c) culturing the adventitious shoots.
Claim 14
A method of making transgenic Acacia mangium plants comprising;
a) preparing transgenic Acacia mangium cells by preculturing stem pieces in culture medium comprising thidiazuron;
b) co-cultivating the stem pieces of step (a) with Agrobacterium in medium comprising thidiazuron to form adventitious buds;
c) culturing the adventitious buds in a selective medium comprising an antibiotic and gibberellic acid; and
d) rooting the plantlets which develop from the adventitious buds of step c).

This application is a divisional of now granted US 6846971 (see above).

WO 2001/53452 A2

  • Earliest priority – 19 January 2000
  • Filed – 19 January 2000
  • OPI – 7 November 2002
Title – Regeneration and genetic transformation of acacia mangium

Claim 1

A method for regenerating Acacia mangium comprising:

A) inducing callus formation from an explant;
B) culturing said callus to produce adventitious buds;
C) culturing said adventitious buds to elongate and produce pinnate leaves; and
D) culturing elongated buds of step (c) such that they produce roots and become plantlets.

Claim 11

A method for regenerating Acacia mangium comprising:

A) culturing auxiliary buds from an Acacia mangium tree to produce adventitious buds comprising phyllodes;
B) subculturing said adventitious buds comprising phyllodes to produce adventitious shoots; and
C) culturing said adventitious shoots.

Claim 14*

A method of transforming Acacia mangium with a gene of interest comprising the steps of:

A) activating Agrobacterium tumefaciens comprising said gene of interest to form activated Agrobacterium tumefaciens;
B) preculturing an explant of Acacia mangium to yield a precultured explant;
C) co-cultivating said activated Agrobacterium tumefaciens and said precultured explant to produce infected explants;
D) culturing said infected explants to induce callus and adventitious buds; and
E) culturing said callus or adventitious buds on a selective medium.

Claim 25

A method for promoting root formation from transformed adventitious buds comprising culturing transformed adventitious buds on a medium comprising 1/2 MS basic medium supplemented with:

A) cx-naphthaleneacetic acid,
B) kinetin,
C) casein enzymatic hydrolysate,
D) L-ascorbic acid,
E) L-glutamine,
F) L-asparagine,
G) L-proline,
H) sucrose and
I) phytagel.

Claim 28*A method of preparing transgenic Acacia mangium cells comprising the steps of:

a) preculturing stem pieces of Acacia mangium in a culture medium; and
b) co-cultivating said stem pieces of step (a) with activated Agrobacterium tumefaciens.

Claim 41A transgenic Acacia mangium cell.
Claim 42A transgenic Acacia mangium plant.

Methods for transforming explants of A. mangium with a gene of interest by co-cultivating A. tumefaciens having the gene of interest with pre-cultured explants of the tree species. In one of the methods the tissue selected for transformation is stems. Transgenic cells and plants of A. mangium are also part of the recited invention.

Additional claims recite methods for the regeneration of non-transformed A. mangium from different tissues.

Institute of Molecular Agrobiology

Remarks
  1. National phase entries of WO 2001/53452 in Canada (CA 2362767) and China (CN 1378599) are pending.
  2. National phase entry of WO 2001/53452 in Europe (EP 1272608) is deemed to be withdrawn on 9 March 2005.
  3. National phase entry of WO 2001/53452 in New Zealand (NZ 514910) has been granted on 27 February 2004.
  4. Other national phase entry of WO 2001/53452 include: Norway (NO 20014331; application rejected according to INPADOC), Brazil (BR 200009112).

Note: Patent information on this page was last updated on 10 March 2006.