*4.5. Cell Wall Isolation*

The plant material (a pool of foliage leaves of 10 plants for each replicate) was powdered by homogenizing the samples in liquid nitrogen with a mortar and pestle. Three g of fresh weight powder were then treated with 50 mL 70% ethanol (*v*/*v*) for 24 h, centrifuged to remove the supernatant (×2), and then incubated with 50 mL 80% acetone (*v*/*v*) for 24 h (×2). The insoluble residues were incubated with 50 mL 2.5 U mL−<sup>1</sup> <sup>α</sup>-amylase (Sigma type VI-A) in 0.01 M phosphate buffer pH 7.0 for 24 h at 37 ◦C (×2). Then, the

remaining pellets were treated with 50 mL phenol-acetic acid-water (2:1:1 *v*/*v*/*v*) for 16 h at room temperature, with a change of solvents after 8 h of incubation. Finally, 50 mL 70% ethanol (*v*/*v*) (×3) and 50 mL 100% acetone (*v*/*v*) (×3) were sequentially added to wash the samples, and the final air-dried residues were considered CWs [61].

## *4.6. FTIR Spectroscopy*

Crude CWs were used to obtain the FTIR spectra using a Jasco 4700 instrument (Tokyo, Japan). The average FTIR spectra (*n* = 10), from 800 to 1800 cm−1, were normalized and baseline-corrected with an ATR module of 4 cm−<sup>1</sup> resolution, using Spectra Manager version 2 (2016) software by Jasco corporation (Tokyo, Japan). A Principal Component Analysis was carried out from normalized spectra with R [101], and visualized with the FactoMineR package [102].

## *4.7. Sequential Polysaccharide Extraction*

The sequential polysaccharide extraction followed the protocol previously described by Rebaque et al. (2017) [61] with slight modifications. Crude CWs (10 mg CW/mL) were treated with cyclohexane-trans-1,2-diamine-*N,N,N ,N* ,-tetraacetic acid sodium salt (CDTA) at pH 6.5 for 8 h. After centrifugation, the pellets were washed with distilled water. The residues obtained were treated with 0.05 M Na2CO3 + 0.02 M NaBH4 and washed with distilled water, to obtain the Na2CO3 fractions. The residues obtained after centrifugation were then incubated in 0.1 N KOH + 20 mM NaBH4 for 8 h, and washed with distilled water, obtaining the KI fractions. For the KII fractions, the remaining pellets were treated with 4 N KOH + 20 mM NaBH4 for 8 h and washed with distilled water. KI and KII fractions were acidified to pH 5.0 with pure glacial acetic acid, and after that, the samples were agitated and centrifugated. Water was added to the pellets obtained after KII fraction, and then were acidified to pH 5 by adding pure glacial acetic acid. The extracts and their respective washings were combined. After its centrifugation, the supernatants were collected to form the Supernatant-Cellulose Residue (SnCR) fractions. The remaining residues were hydrolyzed with 3 mL 2N trifluoroacetic acid (TFA) for 2.5 h at 120 ◦C, centrifuged, and clarified supernatant was referred as TFA fraction.
