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Genetically engineered duckweed

A duckweed and genetic technology, applied in genetic engineering, plant genetic improvement, gardening tools/equipment, etc., can solve problems such as system optimization of culture medium components

Inactive Publication Date: 2000-11-08
NORTH CAROLINA STATE UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

To date, no systematic optimization of media composition and culture conditions has been performed for maximum growth and maximum protein content of specific duckweed strains

Method used

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  • Genetically engineered duckweed

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0088] Eighteen combinations of the auxin 2,4-dichlorophenoxyacetic acid (2,4-D) and the cytokinin benzyladenine (BA) were tested for callus induction in the duckweed species Lemna gibba G3 role.

[0089] Before the experiment, duckweed leaves were cultured at 23°C for 2 weeks in liquid Hoagland medium (Hoagland and Snyder, Proc.Amer.Soc.Hort.Sci.30, 288 (1933)) containing 3% sucrose, photoperiod 16hr light / 8hr dark, the light intensity is about 40μmol / m 2 sec. To induce callus, prepare 18 aliquots of 100 ml Murashige and Skoog basal medium containing 3% sucrose, 0.15% Gelrite and 0.4% Difco Bacterial Agar with 2,4-D concentrations of 10, 20 and 50 μM, And the concentration of BA was 0, 0.01, 0.1, 1.0, 2.0 and 10.0 μM. The pH of all culture media was adjusted to 5.8, autoclaved at 121° C. for 20 minutes, cooled, and each 100 ml was poured into four 100 mm×15 mm petri dishes.

[0090] A full factorial experimental design of 3 2,4-D concentrations × 6 BA concentrations was u...

Embodiment 2

[0093] 40 concentrations of the auxin 2,4-dichlorophenoxyacetic acid (2,4-D) and the cytokinin benzyladenine (BA) were tested to optimize the auxin and cytokinin concentrations by Induced callus from duckweed leaves of Lemnagibba G3.

[0094] Before the experiment, duckweed leaves were cultured at 23°C for 2 weeks in liquid Hoagland medium containing 3% sucrose, with a photoperiod of 16 hr light / 8 hr dark and a light intensity of about 40 μmol / m 2 sec. To induce callus, prepare 40 aliquots of 100 ml Murashige and Skoog medium containing 3% sucrose, 0.15% Gelrite and 0.4% Difco Bacterial Agar with 2,4-D concentrations of 20, 30, 40, 50 , 60, 70, 80, 100 μM, while the concentrations of BA were 0.01, 0.05, 0.1, 0.5 and 1.0 μM. The pH of all culture media was adjusted to 5.8, autoclaved at 121° C. for 20 minutes, cooled, and each 100 ml was poured into four 100 mm×15 mm petri dishes.

[0095] A full factorial experimental design of 8 2,4-D concentrations × 5 BA concentrations w...

Embodiment 3

[0098] Forty combinations of the auxin dicamba and the cytokinin benzyladenine (BA) were tested to compare the relative efficacy of dicamba versus 2,4-D for callus induction of duckweed species Lemna gibba G3.

[0099] Before the experiment, duckweed leaves were cultured at 23°C for 2 weeks in liquid Hoagland medium containing 3% sucrose, with a photoperiod of 16 hr light / 8 hr dark and a light intensity of about 40 μmol / m 2 sec. To induce callus, prepare 40 aliquots of 100 ml Murashige and Skoog medium containing 3% sucrose, 0.15% Gelrite and 0.4% Difco Bacterial Agar with dicamba concentrations of 10, 20, 30, 40, 50, 60, 80, 100 μM, while the concentrations of BA were 0.01, 0.05, 0.1, 0.5 and 1.0 μM. The pH of all culture media was adjusted to 5.8, autoclaved at 121° C. for 20 minutes, cooled, and each 100 ml was poured into four 100 mm×15 mm petri dishes.

[0100] A full factorial experimental design of 8 dicamba concentrations x 5 BA concentrations was used (40 treatments...

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Abstract

Methods and compositions for the efficient transformation of duckweed are provided. Preferably, the methods involve transformation by either ballistic bombardment or Agrobacterium. In this manner, any gene or nucleic acid of interest can be introduced and expressed in duckweed plants. Transformed duckweed plants, cells, tissues are also provided. Transformed duckweed plant tissue culture and methods of producing recombinant proteins and peptides from transformed duckweed plants are also disclosed.

Description

[0001] This invention was made with government support under Grant No. R823570-01-1 from the US Environmental Protection Agency. The government has certain rights in this invention. field of invention [0002] The present invention relates to methods and compositions for transformation of duckweed, in particular transformation methods using ballistic bombardment and Agrobacterium. Background of the invention [0003] Duckweed is the only member of the monocot family Lemnaceae. The four genera and 34 species are small, free-floating freshwater plants whose geographic range spans the entire globe. Landolt, Biosystematic Investigation on the Family of Duckweeds: The Family of Lemnaceae - A Monograph Study. Geobatanischen Institut ETH, Stiftung Rubel, Zurich (1986). Despite being the smallest known plants, most duckweed species have all the tissues and organs of much larger plants, including roots, stems, flowers, seeds and leaves. ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/09A01H5/00C12N5/10C12N15/82C12P21/00
CPCC12P21/00C12N15/8205C12N15/8207
Inventor A·M·斯托普N·拉汉达里
Owner NORTH CAROLINA STATE UNIV
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