Correction: 17 November 2017

www.sciencemag.org/content/357/6356/1118/suppl/DC1

Supplementary Materials for

Biological fabrication of cellulose fibers with tailored properties
Filipe Natalio,* Regina Fuchs, Sidney R. Cohen, Gregory Leitus, Gerhard Fritz-
Popovski, Oskar Paris, Michael Kappl, Hans-Jürgen Butt
*Corresponding author. Email: filipe.natalio@weizmann.ac.il

Published 15 September 2017, Science 357, 1118 (2017)

DOI: 10.1126/science.aan5830

This PDF file includes:

Materials and Methods

Figs. S1 to S18
Table S1
References

Other Supplementary Material for this manuscript includes the following:

(available at www.sciencemag.org/content/357/6356/1118/suppl/DC1)

Movies S1 and S2

Correction: An Editorial Expression of Concern was posted on 14 September 2017, once the editors were
informed of errors in the labeling and description of the control experiments described in figs. S1 and S2.
In particular, in fig. S1, carmine should have been kermesic acid, and in the caption of fig. S2, carmine
should have been carminic acid. After discussions with the authors and minor revisions to the manuscript
and supplementary materials, including corrections to a few other minor errors in the text that do not affect
the conclusions of the paper, the editors are now confident in the results. The Editorial Expression of
Concern has therefore been removed from the PDF of the paper. The paper and supplementary materials
have been corrected.
Materials and Methods

Cotton growth conditions and fertilized ovule culture.

Cotton (Gossypium hirsutum) was hydroponically grown in a greenhouse under
controlled conditions of light, temperature and humidity (10klux, 25°C, 40%) as described
elsewhere (21). Flowers were harvested at 2 day post anthesis (2dpa) and the ovary sterilized in a
solution of 6%NaOCl solution for 10min at room temperature. The fertilized ovules were
aseptically removed and placed floating onto sterile Beasley and Ting (BT) medium (50mL) (18,
19) supplemented with gibberellic acid (GA, 5µM, Sigma, Germany) and indoleacetic acid (IAA,
5µM, Germany). The cultures were kept in the dark at 32°C and 5% CO2 for 20 days.

Synthesis of 6-carboxyfluorescein-glucose (6CF-Glc).

For the synthesis of 6-carboxyfluorescein-glucose (6CF-Glc) conjugate, 50mg of 6-
carboxyfluorescein (116µmol, 1eq., Novabiochem, Germany) were dissolved in dried DMF
(3mL). After, 20 mg of N,N,N′,N′-tetramethyl-O-(N-succinimidyl)uronium tetrafluoroborate
(128µmol, 1.1 eq., TSTU, Sigma, Germany) were added together with 100µL of N,N-
diisopropylethylamine (DIPEA, ReagentPlus®, ≥99%, Sigma, Germany) and left to activate for
30min at room temperature under inert conditions. In parallel, 25mg of 1,3,4,6-tetra-O-acetyl-2-
amino-2-deoxy-β-D-glucopyranose hydrochloride (1 eq., 11µmol, Sigma-Aldrich, Germany)
were dissolved in dried DMF with DIPEA (100µL). The 1,3,4,6-tetra-O-acetyl-2-amino-2-deoxy-
β-D-glucopyranose solution was combined with activated 6-carboxyfluorescein and left to react
overnight at room temperature under inert and dark conditions yielding an orange solid. The
excess of coupling reagents was removed by column chromatography (silica) and elution
followed by thin-layer chromatography (dichloromethane/MeOH 10:1). Deacetylation was
performed by adding a solution of NaOH (3mL, 0.01M) to 15mg of orange conjugate. The
solution was ultra-sonicated for 5s, left to react for 10min at room temperature. The extent of
deacetylation followed by thin-layer chromatography (dichloromethane/MeOH 10:1). To stop the
reaction, a solution of HCl (3mL, 0.01M) was added to neutralize the mixture to pH7.0. The
solvents were retrieved under high vacuum. The product was characterized by UV-Vis, 1H NMR
and mass spectrometry.

Synthesis of Glc-DOTA-Dy(III).
a, the direct coupling from glucosamine hydrochloride to 1,4,7,10-
tetraazacyclododecane-1,4,7,10-tetraacetic acid mono-N-hydroxysuccimide ester (DOTA-NHS)
was performed. Briefly, equimolar amounts of glucosamine hydrochloride (Merck, Germany)
were mixed with DOTA-NHS (CheMatech, France) and DIPEA (1.5eq.) in dry DMSO (1.5mL)
and left to stir overnight at room temperature. The solvent was evaporated under reduced
pressure and the product characterized by 1H and 13CNMR and mass spectrometry. b,
Complexation of Dy(III) with Glc-DOTA was performed as described elsewhere (28). An
aqueous solution of Glc-DOTA (50mg, 3mL) was neutralized to pH7 and equilibrated at 60°C
for 30min before complexation. Afterwards, a solution of DyCl3.7H2O (3eq., 99.99%, Aldrich,
Germany) adjusted to pH7.2 was added to the mixture and stirred for 4h at 60°C, while keeping
the pH at 7 by controlled addition of NaOH (0.02M). The solution was then brought to pH 10,
inducing the precipitation of free dysprosium ions as Dy(OH)3. The solution was brought to
pH7.0, centrifuged at 6000xrpm for 10min at room temperature and the supernatant retrieved and
dried under high vacuum. The free Dy(III) content was assessed by the xylenol orange detection

2
method (29). The protocol was repeated until no free dysprosium ions were found. The Glc-
DOTA-Dy(III) was characterized by mass spectrometry and magnetic measurements (SQUID).
The complexation of Dy(III) only with DOTA (1:1eq., Sigma, Germany) was carried out in
parallel under the same experimental conditions.

Ovule feeding experiments.

6CF-Glc (1mg/mL) was re-suspended in sterile Beasley and Ting BT medium containing
phytohormones (5µM of GA and IAA) (18, 19). The cultures were kept in the dark at 32°C and
5% CO2 for 20 days. Time lapse images (one image per hour) of the cultures were obtained
inside the incubator using homemade construct consisting of a microscope webcamera connected
to a Raspberry pi microcontroller, a customized imaging acquisition software and 1W high
power LEDs (white light and UV λem=365nm). 6-carboxyfluorescein (1mg/mL, Sigma,
Germany), indigo (1mg/mL, 95%, Sigma, Germany), kermesic acid (1 mg/mL, Carbosynth Ltd,
UK), carminic acid (1mg/mL, 95%, Sigma, Germany), 6CF-Glc (1mg/mL), DOTA-Dy(III)
(1mg/mL) and Glc-DOTA-Dy(III) (1mg/mL) were added to IAA-GA supplemented BT medium
and incubated under the same experimental conditions (32°C, 5%CO2 for 20 days). All fibers
were carefully removed with a help of a sterile scalpel and further characterized.

Cryo-sectioning and confocal laser scanning microscope (CLSM) imaging.

All fertilized ovules were allowed to develop under standard incubation conditions (32°C
and 5%CO2 for 20 days). Afterwards, the ovules were harvested, directly immersed in OCT
(Tissue-Tek; Sakura Finetek USA) and immediately frozen at -20°C. Cryo-sections with
thickness of 30μm were prepared using a cryomicrotome (Microm HM 560, Thermo Scientific,
Germany), placed onto previously cooled poly-L-lysine glass slides (VWR, Germany) and kept
at -20°C until further use. The cryo-sections directly imaged using confocal laser scanning
microscope (CLSM, λem = 490nm, Zeiss LSM 710-NLO, Carl Zeiss, Germany). Fibers
autofluorescence was used as background. The images were processed using Zen 2009 software
(Service Pack, Build 5.5.0.443, SMT 5.5.3.1 Carl Zeiss, Germany).
ATR FT-IR, peak fitting&deconvolution.
Infrared analysis of samples was performed using an Agilent Cary 620 spectrometer
(Agilent, Germany) with Focal Plane Array/Mercury cadmium telluride (MCT) detector
fitted with a GladiATR™ (PIKE, USA) connected to a PC for data acquisition. Spectra were
recorded at 4cm-1 resolution, averaging 32 scans. When required, spectra were deconvoluted
in the range 2800-3600cm-1 (OH stretching region) using PeakFit v4.12 (SeaSolve Software
Inc., SYSTAT Software Inc., USA) applying a Gaussian fitting function. Measurements
were performed in triplicate at room temperature.

Scanning Electron Microscopy (SEM).

The samples were fixed onto a carbon tape (Plano GmbH, Wetzlar, Germany) and
analyzed in a scanning electron microscope (SEM) (JEOL JSM-6710F, JEOL Germany GmbH,
Germany) using 2kV acceleration voltage and 2.5x10-6mbar (chamber vacuum).

Differential scanning calorimetry (DSC).

For differential scanning calorimetry (DSC) measurements, the samples were placed in
sealed aluminum crucible and measured in a Netzsch F1 Phoenix (Netzsch-Gerätebau GmbH,
Germany) with the following conditions: air flow (30mL/min) with heating rates of 10K/min up

3
to 800°C. For the buoyancy correction a baseline was recorded using roughly 40mg of dry Al2O3
and subtracted from the sample measurements. The data was analyzed using OriginLab 8
(v8.0725, OriginLab Corporation, USA) and the experiments performed in triplicate.

1
H and 13C NMR and mass spectrometry.
NMR spectra were recorded using the Varian spectrometers Gemini 200, Gemini 2000 or
Unity 500 (δ given in ppm, J in Hz). Data was processed using MestRENova (v. 6.2.1.,
MestRElab Research, Spain). Mass spectra were taken on an Intectra GmbH AMD 402 (electron
impact, 70eV) or on a Thermo Electron Finnigan LCQ (electrospray, voltage 4.5kV, sheath gas
nitrogen) instrument.

Diffuse reflectance UV-Vis (DRUV) spectrophotometry.

Dried cotton raw fibers and FGIO fibers were assembled between two glass slides and
measured in wavelength range between 200 and 700nm with 2nm resolution using JASCO V-670
(JASCO, Germany). The K/S vs. wavelength plot was obtained by applying the Kubelka-Munk
function (F(R∞)) (eq. 1)

F(R∞)= (1-R∞)2 = K (eq. 1)

2R∞ S

Where R∞ is the reflectance of an infinite thick layer, K is the absorption coefficient and
S the scattering coefficient. BaSO4 was used as “white standard”. For the calibration curve,
silica nanoparticles (diameter 20nm, Ludox 40, Sigma, Germany) and silica nanoparticles
mixed with different concentration of 6CF-Glc (0.8, 2.5, 5, 7.5 and 10%), assembled
between two glass slides and sealed with scotch tape. The measurements were performed in
triplicate (N=3). The data was treated using OriginLab 8 (v8.0725, OriginLab Corporation,
USA).

Sample digestion and ICP-MS.

Control sample (raw fibers) and fibers co-incubated with Glc-DOTA-Dy(III) and DOTA-
Dy(III) were wetted in 1mL distilled water and then digested in concentrated nitric acid (HNO3,
2.4mL, 70%, ≥99.999% trace metals basis, Aldrich, Germany) and HCl (7.2mL, 30-35%,
TraceSELECT®Ultra, for ultratrace analysis, Fluka, Germany), using a microwave oven (model
microPREP-A, Germany). The digestion process included multiple programs; 6min at 1000W
(160°C), 9min at 1000W (195°C), 20min at 1000W (195°C). The obtained solutions were placed
in volumetric flasks after cooling and diluted with distilled water to final volume of 50mL. For
the calibration curve, solutions of dysprosium (dysprosium chloride hexahydrate, DyCl3.7H2O, ≥
99.99% trace metals basis, Aldrich, Germany), with five different concentrations (0, 11, 50, 110,
560 and 1000ppb) were prepared in distilled water. As control, distilled water was used. All
samples were prepared in triplicate (N=3), treated as above and measured. A quadrupole based
mass spectrometer (Plasma Quant MS Elite, Analytik Jena, Germany) was used for the ICP-MS
measurements and the data acquired on an ASpect MS software (Analytik Jena, Germany). The
data was treated using OriginLab 8 (v8.0725, OriginLab Corporation, USA).

4
Atomic Force Microscopy (AFM).
Individual raw fibers, FGIO fibers and FiDy fibers were carefully separated from the
cotton ovules after growth under standard conditions. Each fiber was gripped at both ends with
glue and parallel to the Si wafer surface. AFM measurements were made as close as possible to
the epoxy attachment point but without touching the glue itself. The AFM (Bruker Multimode)
was operated in both tapping mode, and peak-force QNM for topography using Olympus A160
probes.

Small angle X-ray scattering (SAXS) and wide angle X-ray scattering (WAXS).
SAXS: Bundles of raw fibers, FGIO fibers and FiDy fibers were measured on a NanoStar
instrument (Bruker-AXS, Germany), equipped with a IµS microsource and a Vantec-2000
detector at wavelength λ = 0.15418nm. The sample-detector distance of 1070mm and the centre
of the primary beam were determined using silver behenate as a calibration standard. The
exposure time during each experiment was 600s. The samples were measured at several
positions, twice at each position. Azimuthal intensity profiles I(χ) were obtained from the 2D
SAXS patterns (fig. S8) by averaging all intensities within the q-range 0.5<q<1nm-1, where
q=4πsin(θ)/λ, 2θ being the scattering angle. Azimuthal integration was performed within an
angular slice of one degree. Radial intensity distributions I(q) were obtained from slices of 20°
angular opening in the direction of the fiber axis and normal to it.
WAXS: Bundles of raw fibers, FGIO fibers and FiDy fibers were measured using a Bruker
Nanostar (Bruker AXS, Germany) using CuKα radiation. An imaging plate at a distance of
53mm was used as a detector. Exposure time was 10min. Radial integration of the images was
done in an angular range of +/-15° around equatorial direction. The range
0.189Å-1 < q < 3.39Å-1 for WAXS was divided in 1200bins, with the magnitude of the
scattering vector defined in the same way as for SAXS. Azimuthal integration was done
within the range 1.00Å-1 < q < 1.75Å-1.

Tensile testing.
Individual raw, FGIO fibers and FiDy fibers were carefully separated from the cotton
ovules after growth under standard conditions. Each fiber was gripped at both ends over a hole
with a diameter of 2mm on card templates, where small spots of glue had been placed at opposite
sides. Each end of a fiber was brought in contact with the glue spot, while the central fiber
section was loose and not under tension. This allowed us to orient the fiber ends in parallel
direction before the glue hardened. The card template was mounted on a holder and the card
material was cut on one side only under the video microscope with a sharp scissor. A wedge like
structure was employed for one side of the template to avoid contact forces between the card
edges. This cut-out also later allows for more freedom to realign the fiber to ensure vertical
loading direction. The half-cut template was mounted on a homebuilt Force-Puller (fig. S11)
using appropriately designed template grips. Then the second half of the template was cut with a
scissor under a video microscope. To correct for any possible offset that may have occurred
during mounting or cutting of the template, we could manually shift the lower holder in the x-y
direction under the video microscope. As the fibers were not under tension during any of these
preparation steps, the possible slight deformation of the card material during cutting should not
lead to strain and possible damage of the fibers. In fact, in all tensile tests we had to stretch the
fibers for some distance with the help of the stepper motor before a measurable onset of stress

5
was detected by the force sensor. The data was treated using OriginLab 8 (v8.0725, OriginLab
Corporation, USA).

Magnetic measurements (SQUID).

Magnetic moment of the raw fibers (6.7mg) and FiDy fibers (3.2mg) were measured
at SQUID magnetometer MPMS3 (LOT-Quantum Design Inc., USA) by using VSM mode.
Field cooled temperature dependences of magnetic moment M(T) were taken during sample
heating at the magnetic field strength H=5kOe and H=60kOe in the temperature range from
T=2K to T=300K, 1K steps. Field dependences of magnetic moment M(H) were obtained at
the temperatures T=2K and 300K when the field was decreased and increased in the interval
-6T≤H≤+6T. The data was treated using OriginLab 8 (v8.0725, OriginLab Corporation,
USA).

6
Fig. S1. G. hirsutum fertilized ovules incubated with pigments devoid from glucose and
under standard growth conditions (32°C, 5%CO2, 20 days). Left column: representative
optical microscopic images of fully grown (A) indigo; (B) kermesic acid (1mg/mL) and (C)
control. No coloration or fluorescence is observed for the fibers of any of the samples.
Right column: correspondent representative microscopic images of cryo-sections. No
coloration is observed at the epidermal cells suggesting that the uptake of exogenous
molecules is regulated by an active transport system (e.g. glucose transporters).

7
Fig. S2. (A) Representative microscopic image of G. hirsutum fertilized ovules cryogenic
cross-sections after incubation with carminic acid (1mg/mL). The experiments were
performed under standard conditions (32°C, 5% CO2, 20 days). A clear red coloration inside
the epidermal cells (accumulation) of carminic acid is observed (black arrows). (B and C)
Digital camera image of the fertilized ovule grown in the presence of carminic acid
(1mg/mL) (B) and standard condition (C). No red coloration of the fibers is observed.

8
Fig. S3. Schematic representation of synthesis of 6-carboxyfluorescein-glucose (6CF-Glc)
starting from 6-carboxyfluorescein and 1,3,4,6-tetra-O-acetyl-2-amino-2-deoxy-β-D-
glucopyranose. Inset: UV-Vis spectra measured in distilled water, pH 6. Black line: 6-
carboxyfluorescein with bands at 228/448/471nm. Red Line: 6CF-Glc with bands at
230/454/477nm. The observed peak slight redshift infers glucose conjugation to the 6-
carboxyfluorescein. Yield: 89%. 1H NMR (400 MHz, CD3OD) δ (ppm): 8.55 (d, J = 5.9 Hz,
11H), 8.05 ñ 7.84 (m, 39H), 7.70 (s, 6H), 6.82 ñ 6.65 (m, 22H), 5.48 (s, 6H), 4.34 (s, 465H),
3.71 (s, 5H), 3.60 (q, J = 7.0 Hz, 148H). ESI-MS (m/z, CH3CN/formic acid 0.01%):
expected 538.73[M-H]+, found 621.36 [M-2CH3CN-H]+.

9
Fig. S4. (A) Digital image of the homemade setup inside the incubator. This setup allows
real time observation of development of cotton fibers in the presence of exogenous
molecules under “white” and UV (365nm) light. (B) Schematic representation. Component
specification (camera, high power LED`s, raspberry pi), python code and *.stl files are
available on request.

10
Fig. S5. Sequence of images (acquired hourly) showing fiber development of G. hirsutum
fertilized ovules in the absence of any exogenous molecules (control). The experiments were
performed under standard conditions (32°C, 5%CO2, 20 days). The images were obtained
using our home built system described in Fig. S4. (A) “white” light. (B) UV light (λ=365nm).

11
Fig. S6. (A) K/S vs.wavelength plot (350-700nm, 2nm resolution) of air-dried and fibers
excised from G. hirsutum fertilized ovules in the absence (control, black line) and in
presence 6-carboxyfluorescein-glucose (red line) after growing under standard growth
conditions (32°C, 5%CO2, 20 days). (B) Standard curve of different 6-carboxyfluorescein-
glucose conjugate/silica nanoparticles in different concentrations (0.8, 2.5, 5, 7.5 and 10%).
Each point is an average of three points (N=3) and obtained from the maximum value of the
band at 496nm. Linearization was obtained using OriginLab 8 (v8.0725, OriginLab
Corporation, USA) and the uptake determined to be 4.75+/-0.2%.

12
 A B C

Fig. S7. 2D SAXS patterns of (A), raw fibers; (B), fibers containing 6-
carboxylfluorescein-glucose (FGIO fibers); (C), fibers containing Glc-DOTA-Dy(III)
(FiDy fibers). The black lines indicate fiber orientation.

13
Fig. S8. Azimuthal SAXS intensity profiles from of the aligned fibers (blue line=raw
fibers, red line=FGIO fibers, green line=FiDy fibers).

14
Fig. S9. Radial cuts through the SAXS patterns of the aligned fibers. Dotted curves are cuts
parallel to the fiber orientation, solid lines are normal to the fiber axis (blue line=raw fibers,
red line=FGIO fibers, green line=FiDy fibers).

Note: Cracks due to fiber drying might also cause an anisotropic SAXS signal due to the
oriented surface scattering. This however, is unlikely to cause the observed intensity pattern
since such surface scattering effects caused by the fractures should decay faster with
increasing scattering angle than observed (30). Additionally, the radial scattering profiles of
the modified cellulose fibers are very similar to the ones obtained for unmodified cotton and
resemble the well-known shape of SAXS from cellulose nanofibers in plants (31). All the
curves also show features that cannot be explained by a simple power law as one would
expect for surface scattering of macroscopic fractures. Therefore, oriented fibrillary
structures with typical diameters of a few nanometres are present within the fibers.

15
Fig. S10. Representative SEM images of (A) raw fibers (average thickness 1.7+/-0.4µm/average
width 21+/-5µm. N=20); (B) fibers containing 6-carboxylfluorescein-glucose (FGIO) (average
thickness 1.5+/-0.3µm/average width 24+/-5µm. N=20); (C) fibers containing Glc-DOTA-
Dy(III) (FiDy fibers) (average thickness 1.0+/-0.3µm/average width 18+/-6µm. N=20).

16
Fig. S11. (A) Digital image of the homebuilt Force-Puller-Apparatus (FPA). This setup was
designed for the purpose of measuring the tensile strength of individual cotton fibers. (B)
Designed card template with a fiber glued on top crossing the central hole (diameter of
2mm). The sides of the card template were cut after mounting to release the fiber before
testing. (C) Set-up scheme: The card template (1) was mounted between a strain gauges force
sensor (2) on top of a motorized Z-stage (3) and a fixed counter-holder (4). The deflection of
the load sensor during pulling of the upper half of the card template changes the resistivity of
the integrated thin film resistor, which was recorded using a Wheatstone bridge and a
preamplifier. The force sensor was calibrated by adding defined masses to the end of the
sensor and recording the corresponding output signal of the preamplifier. A calibration factor
of ~12.36mN/V was used to convert all force signals [V] into tensile force in [mN]. The
maximum force range is 98mN at a noise level of the whole detection system of less than
75µN. The motorized Z-stage (HVM130-30-HDS, Owis, Staufen, Germany, minimum step
size 8nm) was controlled by a computer that was also equipped with an AD/DA board (NI
PCI-6251, National Instruments Corporation, Austin TX, USA) running an in-house
LabVIEW program. It allows automated acquisition of the tensile force-displacement curve.
The motorized Z-stage was moved upwards to stretch the fiber until failure of the fiber
occurs. The stretching and rupture of the fiber were recorded by a video microscope (5).

17
Fig. S12. Representative sequence of images showing the tensile testing of raw fibers
probed with a Force-Puller setup and recorded in situ with a microscopy camera (i to vi)
(scale bar: 60µm). The fibers are stretched until fracture (vi).

18
Fig. S13. Schematic representation of Glc-DOTA-Dy(III) synthesis starting from a. DOTA-
NHS and glucosamine hydrochloride (1:1, dry DMSO, RT, overnight). Yield: 93%. 13C
NMR (126 MHz, D2O) δ 176.33 (s), 170.24 (s), 94.44 (s), 71.57 (s), 70.76 (s), 60.88 -
65.42 (m), 60.50 (dd, J = 28.3, 18.7 Hz), 59.13 (s), 54.30 (d, J = 7.4 Hz), 54.02 (s). 1H
NMR (500 MHz, D2O) δ 8.11 (s), 6.34 (d), 4.65 (s, 78H), 4.00 - 4.53 (m, 85H), 3.94 (s,
4H), 3.30 (s, 8H), 3.25 (s, 23H), 2.46 (s, 3H). ESI-MS (m/z, CH3CN/formic acid 0.01%):
expected 565.6 [M], found 588.52[M-Na+]. b. further complexed with Dy(III) (pH 10.65,
80°C for 4h). Yield: 85%, ESI-MS (positive mode, m/z, CH3CN/formic acid 0.01%):
expected 725.37 [M], found 749.15 [M-Na+] and 386.17 [M-2Na+]. Control experiments
for complexation of DOTA with Dy(III) under the same experimental conditions. ESI-MS
(m/z, CH3CN/formic acid 0.01%): expected 565.26 [M], found 565.22 [M].

19
Fig. S14. DC magnetic measurements for Glc-DOTA-Dy(III) complex. (A) Temperature
dependence of susceptibility magnetic product (χMT) measured at 1kOe. The χMT is 14.12
emu.K.mol-1 at 300K, in good agreement with the theoretical values for a single Dy(III) ion
(6H15/2, S=5/2, L=5, J=15/2, g=4/3) (14.17emu.K.mol-1) (27). The susceptibility product
steadily decreases to reach 11.04cm3.K.mol -1 at 2K indicative of paramagnetic behavior.
This decrease is attributed to crystal-field effects, i.e., thermal depopulation of the Stark
sublevels of Dy(III) ion of the 6H15/2 ground state. The values found at low temperature and
weak field suggest negligible magnetic interactions, a typical behavior found in single
molecular magnets (SMM). (B) M vs.H plots shows the presence of magnetic anisotropy,
due to the incomplete saturation of the magnetization. The magnetization at 2K and 60kOe
is 5.03µB, which is closed to the expected value for one Dy(III) ion (5.23µB).

20
Fig. S15. (A) Digital camera images of the G. hirsutum fertilized ovules grown under
standard conditions (32°C, 5%CO2, 20 days) incubated in the absence of any exogenous
molecule (control, left ovule) and with DOTA-Dy(III) (1mg/mL) (right ovule). The images
were taken immediately after leaving the incubator. No major differences are observed. (B)
SEM image of the collected fibers incubated with DOTA-Dy(III) (FiDy fibers) (1mg/mL)
showing the typical flat immature morphology. (C) Magnified SEM image of FiDy fibers.

21
Fig. S16. Atomic force microscopy image (topography) of end-clamped single fiber (on Si
wafer) collected from G. hirsutum fertilized ovules grown under standard conditions (32°C,
5%CO2, 20 days) showing the typical fibrillar texture of the cotton fibers.

22
Fig. S17. (A) Representative force-displacement curves obtained during tensile testing of a
single fiber isolated from G. hirsutum fertilized ovules grown under standard conditions
(32°C, 5%CO2, 20 days) incubated with Glc-DOTA-Dy(III) (1mg/mL). (B) Representative
SEM image of one single fiber after fracture.

23
Fig. S18. DC magnetic measurements for Glc-DOTA-Dy(III) after subtracting the
diamagnetic contribution of cotton fibers. Temperature dependence of molar magnetic
susceptibility (χ) measured at 5kOe. The χT is 14.12emu.K.mol-1 at 300K, in good
agreement with the theoretical values for a single Dy(III) ion (6H15/2, S=5/2, L=5, J=15/2,
g=4/3) (14.17emu.K.mol-1) (27). The χT steadily decreases to reach 10.97cm3.K.mol-1 at
2K indicative of paramagnetic behavior. This decrease is attributed to crystal-field effects,
i.e., thermal depopulation of the Stark sublevels of Dy(III) of the 6H15/2 ground state (27).
The values are very similar to those found for Glc-DOTA-Dy(III) complex alone. Error
bars are presented on the graphs for all experimental points.

24
Table S1. Counts of the location of fiber rupture for raw fibers, fibers containing 6-
carboxylfluorescein-glucose (FGIO fibers), and fibers containing Glc-DOTA-Dy(III)
(FiDy fibers).

Fiber rupture Raw FGIO Glc-DOTA-Dy(III)

position (counts) (counts) (counts)
Edge 15 19 36
Middle 8 14 10
Total 23 33 46

Edge (%) 65 58 78
Middle (%) 35 42 22

25
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