Fig 1.
Pilocarpus microphyllusStapf ex Wardleworth(jaborandi).
(A) P. microphyllus, highlighting the terminal leaflets (Photo: Anidro do Brasil Extrações S.A). (B) P. microphyllus harvested with pruning shears (Photo: Anidro do Brasil Extrações S.A.).
Table 1.
Primers used for generating polymorphic ISSR markers in Pilocarpus microphyllus genotypes, showing the respective levels of polymorphic loci.
Fig 2.
Chromatograms and mass spectra of alkaloid standards from Pilocarpus microphyllus analyzed by LC/MS.
Pilocarpine, showing the retention time at 15.05 min and molecular weight for mass spectrometry, electron spray [M + H]+ at 209 Da. (B) Epiisopiloturine, showing the retention time at 17.05 min and molecular weight for mass spectrometry, electron spray [M + H] + at 287 Da.
Fig 3.
Rainfall and pilocarpine content (%) in the dry and rainy seasons.
(A) Rainfall distribution (mm) during the dry and rainy seasons [0.00 mm; 109.69 mm (p = 0.0003), respectively] in a P. microphyllus plantation, Piauí state, northeastern Brazil. (B) Percentage of PIL (w/w) by the dry and rainy seasons [0.727; 0.683 (p = 0.5637), respectively] from S01. (C) Percentage of PIL (w/w) by the dry and rainy seasons [0.560; 0.350 (p = 0.0149), respectively] from S02. (D) Percentage of PIL (w/w) by the dry and rainy seasons [0.687; 0.415 (p = 0.0030), respectively] from S03.
Fig 4.
Typical chromatographic profiles obtained by HPLC analysis of the three cultivated populations of P. microphyllus.
S01 group (jaborandi green line). (B) S02 group (jaborandi traditional line). (C) S03 (jaborandi traditional line). Peak no. 1: pilocarpine; peak no. 2: epiisopiloturine.
Fig 5.
Pilocarpine (%, w/w) over one year, measured by HPLC analysis.
Each point represents the mean ± standard deviation of a sample of three leaf extractions.Different letters indicate significant differences between S01, S02, and S03 with Tukey’s test at the 5% level.
Table 2.
Pilocarpine (%, w/w) contents in S01, S02, and S03 over one year, measured with HPLC.
Table 3.
Epiisopiloturine content (%, w/w) in S01, S02, and S03 over one year, measured with HPLC.
Table 4.
Genetic variation of the 15 individuals from the three groups of Pilocarpus microphyllus (S01, S02, S03) analyzed with GenAlEx 6.5 software.
Table 5.
Analysis of molecular variance (AMOVA) of the three groups of cultivated Pilocarpus microphyllus, analyzed by GenAlEx 6.5 software.
Table 6.
Pairwise genetic distances (as PhiPT values) between the three groups (S01, S02, S03) of cultivated Pilocarpus microphyllus, based on ISSR marker data analyzed with GenAlEx 6.5 software.
Fig 6.
Principal coordinate analysis of 11 morphological quantitative variables and 111 ISSR binary molecular markers from three groups of Pilocarpus microphyllus (S01, S02, S03), representing two cultivated lines.
Green line (black: S01). Traditional line (red: S02; blue: S03)