Zingiber ultralimitale, a new species from Sulawesi 
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Material and methods 
Detailed studies of the morphology were made using living collections grown in the 
greenhouse of the Royal Botanic Garden Edinburgh. Measurements were made using 
a ruler and a calibrated eye piece under a dissecting microscope. 
For the DNA barcoding, three barcoding regions were successfully sequenced, 
namely rbcL, trnH-psbA and the nuclear Internal Transcribed Spacer (ITS). The 
fourth barcoding region, matK could not be sequenced. Extraction, amplification and 
sequencing followed Kress et al. (2002) and Kress & Erickson (2007). The material 
sequenced was from Poulsen et al. 2767. GenBank accession numbers for the three 
barcoding regions are summarised in Table 2. A phylogenetic analysis based on ITS 
sequences was performed using additional sequences from NCBI GenBank after 
BLAST was done. GenBank has very few records of rbcL and trnH-psbA so it was 
not possible to run an analysis using these regions to address the affiliation of Zingiber 
ultralimitale. Kaempferiaparviflora Wall, ex Baker and K. elegans (Wall.) Baker were 
chosen as the outgroup following Theerakulpisut et al. (2012) (Table 3). Sequence 
alignment and Maximum Likelihood analysis were conducted using MEGA version 6 
(Tamura et al., 2013). 
Table 2. Voucher information and Genbank accession numbers. 
Species 
Gene region 
Genbank accession 
number 
Material 
(Herbarium location) 
Zingiber ultralimitale 
rbcL 
KU891637 
Poulsen et al. 2767 
trnH-psbA 
KU891638 
(BO, E) 
ITS 
KU891639 
For the palynological study, material was collected from a living plant growing 
at the Royal Botanic Garden Edinburgh (ace. no 20092015 A) and fixed in FAA for 
at least 24 hours. Mature but unopened flowers were selected. For SEM, anthers were 
dissected from the flowers, put into tiny chambers and dehydrated for 45 minutes 
in 50%, 70%, 95%, and 100% ethanol, ten minutes in acetone (two changes of five 
minutes each). Critical point drying was carried out in an Emitech K850 dryer and 
samples were sputter-coated with platinum and examined using a Zeiss Supra 55VP 
SEM. 
Results and discussion 
Most of the Zingiber species of western Malesia have tightly imbricate bracts, long or 
very short peduncles, and lateral staminodes joined to the labellum, although Zingiber 
