prokaryotes. Science 80(315):1709–1712. https://doi.
org/10.1126/science.1138140
Becker S (2006) Immuntoxinproduktion in Pflanzen. Eine
innovative Methode zur Herstellung neuartiger Therapeutika gegen das Hodgkin Lymphom. PHD Thesis
Bernard FA, Bernard JM, Denny P (1990) Flower
structure, anatomy and life history of Wolffia australiana (Benth.) den Hartog and van der Plas. Bull
Torrey Bot Club 117:18–26. https://doi.org/10.2307/
2997125
Bertran K, Thomas C, Guo X et al (2015) Expression of
H5 hemagglutinin vaccine antigen in common duckweed (Lemna minor) protects against H5N1 high
pathogenicity avian influenza virus challenge in
immunized chickens. Vaccine 33:3456–3462. https://
doi.org/10.1016/j.vaccine.2015.05.076
Boehm R, Kruse C, Voeste D et al (2001) A transient
transformation system for duckweed (Wolffia columbiana) using Agrobacterium-mediated gene transfer.
J Appl Bot 75:107–111
Bog M, Schneider P, Hellwig F et al (2013) Genetic
characterization and barcoding of taxa in the genus
Wolffia Horkel ex Schleid (Lemnaceae) as revealed by
two plastidic markers and amplified fragment length
polymorphism (AFLP). Planta 237:1–13. https://doi.
org/10.1007/s00425-012-1777-9
Cantor-Pastor A, Mollar-Morales A, Ernst E et al (2015)
Efficient transformation and artificial miRNA gene
silencing in Lemna minor. Plant Biol 17:59–65.
https://doi.org/10.1111/plb.12215
Cao HX, Vu GTH, Wang W et al (2016) The map-based
genome sequence of Spirodela polyrhiza aligned with
its chromosomes, a reference for karyotype evolution.
New Phytol 209:354–363. https://doi.org/10.1111/
nph.13592
Cheng JJ, Stomp A-M (2009) Growing duckweed to
recover nutrients from wastewaters and for production
of fuel ethanol and animal feed. Clean-Soil Air Water
37:17–26. https://doi.org/10.1002/clen.200800210
Chhabra G, Chaudhary D, Sainger M, Jaiwal PK (2011)
Genetic transformation of Indian isolate of Lemna minor
mediated by Agrobacterium tumefaciens and recovery of
transgenic plants. Physiol Mol Biol Plants 17:129–136.
https://doi.org/10.1007/s12298-011-0059-5
Cong L, Ran FA, Cox D et al (2013) Multiplex genome
engineering using CRISPR/Cas systems. Science 80
(339):819–823. https://doi.org/10.1126/science.1231143
Cox KM, Sterling JD, Regan JT et al (2006) Glycan
optimization of a human monoclonal antibody in the
aquatic plant Lemna minor. Nat Biotechnol 24:1591–
1597. https://doi.org/10.1038/nbt1260
Crespan E, Czabany T, Maga G, Hübscher U (2012)
Microhomology-mediated DNA strand annealing and
elongation by human DNA polymerases k and b on
normal and repetitive DNA sequences. Nucleic Acids
Res 40:5577–5590. https://doi.org/10.1093/nar/gks186
Cui W, Cheng JJ (2015) Growing duckweed for biofuel
production: a review. Plant Biol 17:16–23. https://doi.
org/10.1111/plb.12216
den Hartog C, van der Plas F (1972) The Australian
species of Wolffia (Lemnaceae). Blumea—Biodiv
Evol Biogeogr Plants 20:151–153
Deriano L, Roth DB (2013) Modernizing the nonhomologous end-joining repertoire: alternative and classical
NHEJ share the stage. Annu Rev Genet 47:433–455.
https://doi.org/10.1146/annurev-genet-110711155540
Dhir B, Sharmila P, Saradhi PP (2009) Potential of
aquatic macrophytes for removing contaminants from
the environment. Crit Rev Environ Sci Technol
39:754–781
Dicarlo JE, Norville JE, Mali P et al (2013) Genome
engineering in Saccharomyces cerevisiae using
CRISPR-Cas systems. Nucleic Acids Res 41:4336–
4343. https://doi.org/10.1093/nar/gkt135
Doench JG, Fusi N, Sullender M et al (2016) Optimized
sgRNA design to maximize activity and minimize
off-target effects of CRISPR-Cas9. Nat Biotechnol
34:184–191. https://doi.org/10.1038/nbt.3437
Dushenkov V, Kumar PBAN, Motto H, Raskin I (1995)
Rhizofiltration: the use of plants to remove heavy
metals from aqueous streams. Environ Sci Technol
29:1239–1245. https://doi.org/10.1021/es00005a015
Edelman M, Perl A, Flaishman M, Blumenthal A (1998)
Patent US 7,176,352 B1: Transgenic Lemnaceen,
pp 1–34
Engler C, Kandzia R, Marillonnet S (2008) A one pot, one
step, precision cloning method with high throughput
capability. PLoS ONE 3:e3647. https://doi.org/10.
1371/journal.pone.0003647
Engler C, Youles M, Gruetzner R et al (2014) A golden
gate modular cloning toolbox for plants. ACS Synth
Biol 3:839–843. https://doi.org/10.1021/sb4001504
Fauser F, Schiml S, Puchta H (2014) Both
CRISPR/Cas-based nucleases and nickases can be
used efficiently for genome engineering in Arabidopsis thaliana. Plant J 79:348–359. https://doi.org/10.
1111/tpj.12554
Friedrich AS (2005) Untersuchungen zu Kultivierung.
Universität Bonn, Transformation und Fermentation
von Wolffia spec
Garneau JE, Dupuis MÈ, Villion M et al (2010) The
CRISPR/cas bacterial immune system cleaves bacteriophage and plasmid DNA. Nature 468:67–71.
https://doi.org/10.1038/nature09523
Humann J, Ficklin S, Lee T et al (2017) Structural and
functional annotation of model and non-model organisms with GenSAS v5.0, a web-based annotation
platform. In: Bioinformatics Workshop, International
Plant and Animal Genome Conference XXV. San
Diego, CA, USA
Jiang W, Bikard D, Cox D et al (2013a) RNA-guided
editing of bacterial genomes using CRISPR-Cas
systems. Nat Biotechnol 31:233–239. https://doi.org/
10.1038/nbt.2508
Jiang W, Zhou H, Bi H et al (2013b) Demonstration of
CRISPR/Cas9/sgRNA-mediated targeted gene modification in Arabidopsis, tobacco, sorghum and rice.
17 Editing the Genome of Wolffia australiana
175
org/10.1126/science.1138140
Becker S (2006) Immuntoxinproduktion in Pflanzen. Eine
innovative Methode zur Herstellung neuartiger Therapeutika gegen das Hodgkin Lymphom. PHD Thesis
Bernard FA, Bernard JM, Denny P (1990) Flower
structure, anatomy and life history of Wolffia australiana (Benth.) den Hartog and van der Plas. Bull
Torrey Bot Club 117:18–26. https://doi.org/10.2307/
2997125
Bertran K, Thomas C, Guo X et al (2015) Expression of
H5 hemagglutinin vaccine antigen in common duckweed (Lemna minor) protects against H5N1 high
pathogenicity avian influenza virus challenge in
immunized chickens. Vaccine 33:3456–3462. https://
doi.org/10.1016/j.vaccine.2015.05.076
Boehm R, Kruse C, Voeste D et al (2001) A transient
transformation system for duckweed (Wolffia columbiana) using Agrobacterium-mediated gene transfer.
J Appl Bot 75:107–111
Bog M, Schneider P, Hellwig F et al (2013) Genetic
characterization and barcoding of taxa in the genus
Wolffia Horkel ex Schleid (Lemnaceae) as revealed by
two plastidic markers and amplified fragment length
polymorphism (AFLP). Planta 237:1–13. https://doi.
org/10.1007/s00425-012-1777-9
Cantor-Pastor A, Mollar-Morales A, Ernst E et al (2015)
Efficient transformation and artificial miRNA gene
silencing in Lemna minor. Plant Biol 17:59–65.
https://doi.org/10.1111/plb.12215
Cao HX, Vu GTH, Wang W et al (2016) The map-based
genome sequence of Spirodela polyrhiza aligned with
its chromosomes, a reference for karyotype evolution.
New Phytol 209:354–363. https://doi.org/10.1111/
nph.13592
Cheng JJ, Stomp A-M (2009) Growing duckweed to
recover nutrients from wastewaters and for production
of fuel ethanol and animal feed. Clean-Soil Air Water
37:17–26. https://doi.org/10.1002/clen.200800210
Chhabra G, Chaudhary D, Sainger M, Jaiwal PK (2011)
Genetic transformation of Indian isolate of Lemna minor
mediated by Agrobacterium tumefaciens and recovery of
transgenic plants. Physiol Mol Biol Plants 17:129–136.
https://doi.org/10.1007/s12298-011-0059-5
Cong L, Ran FA, Cox D et al (2013) Multiplex genome
engineering using CRISPR/Cas systems. Science 80
(339):819–823. https://doi.org/10.1126/science.1231143
Cox KM, Sterling JD, Regan JT et al (2006) Glycan
optimization of a human monoclonal antibody in the
aquatic plant Lemna minor. Nat Biotechnol 24:1591–
1597. https://doi.org/10.1038/nbt1260
Crespan E, Czabany T, Maga G, Hübscher U (2012)
Microhomology-mediated DNA strand annealing and
elongation by human DNA polymerases k and b on
normal and repetitive DNA sequences. Nucleic Acids
Res 40:5577–5590. https://doi.org/10.1093/nar/gks186
Cui W, Cheng JJ (2015) Growing duckweed for biofuel
production: a review. Plant Biol 17:16–23. https://doi.
org/10.1111/plb.12216
den Hartog C, van der Plas F (1972) The Australian
species of Wolffia (Lemnaceae). Blumea—Biodiv
Evol Biogeogr Plants 20:151–153
Deriano L, Roth DB (2013) Modernizing the nonhomologous end-joining repertoire: alternative and classical
NHEJ share the stage. Annu Rev Genet 47:433–455.
https://doi.org/10.1146/annurev-genet-110711155540
Dhir B, Sharmila P, Saradhi PP (2009) Potential of
aquatic macrophytes for removing contaminants from
the environment. Crit Rev Environ Sci Technol
39:754–781
Dicarlo JE, Norville JE, Mali P et al (2013) Genome
engineering in Saccharomyces cerevisiae using
CRISPR-Cas systems. Nucleic Acids Res 41:4336–
4343. https://doi.org/10.1093/nar/gkt135
Doench JG, Fusi N, Sullender M et al (2016) Optimized
sgRNA design to maximize activity and minimize
off-target effects of CRISPR-Cas9. Nat Biotechnol
34:184–191. https://doi.org/10.1038/nbt.3437
Dushenkov V, Kumar PBAN, Motto H, Raskin I (1995)
Rhizofiltration: the use of plants to remove heavy
metals from aqueous streams. Environ Sci Technol
29:1239–1245. https://doi.org/10.1021/es00005a015
Edelman M, Perl A, Flaishman M, Blumenthal A (1998)
Patent US 7,176,352 B1: Transgenic Lemnaceen,
pp 1–34
Engler C, Kandzia R, Marillonnet S (2008) A one pot, one
step, precision cloning method with high throughput
capability. PLoS ONE 3:e3647. https://doi.org/10.
1371/journal.pone.0003647
Engler C, Youles M, Gruetzner R et al (2014) A golden
gate modular cloning toolbox for plants. ACS Synth
Biol 3:839–843. https://doi.org/10.1021/sb4001504
Fauser F, Schiml S, Puchta H (2014) Both
CRISPR/Cas-based nucleases and nickases can be
used efficiently for genome engineering in Arabidopsis thaliana. Plant J 79:348–359. https://doi.org/10.
1111/tpj.12554
Friedrich AS (2005) Untersuchungen zu Kultivierung.
Universität Bonn, Transformation und Fermentation
von Wolffia spec
Garneau JE, Dupuis MÈ, Villion M et al (2010) The
CRISPR/cas bacterial immune system cleaves bacteriophage and plasmid DNA. Nature 468:67–71.
https://doi.org/10.1038/nature09523
Humann J, Ficklin S, Lee T et al (2017) Structural and
functional annotation of model and non-model organisms with GenSAS v5.0, a web-based annotation
platform. In: Bioinformatics Workshop, International
Plant and Animal Genome Conference XXV. San
Diego, CA, USA
Jiang W, Bikard D, Cox D et al (2013a) RNA-guided
editing of bacterial genomes using CRISPR-Cas
systems. Nat Biotechnol 31:233–239. https://doi.org/
10.1038/nbt.2508
Jiang W, Zhou H, Bi H et al (2013b) Demonstration of
CRISPR/Cas9/sgRNA-mediated targeted gene modification in Arabidopsis, tobacco, sorghum and rice.
17 Editing the Genome of Wolffia australiana
175