Artabotrys rubriflorus (Annonaceae), a new species from Yunnan China

Artabotrys rubriflorus sp. nov. is described as a new species of Annonaceae, collected from Malipo, Yunnan Province, China.The specimens of this new species were gathered through fieldwork and were subsequently subjected to rigorous analysis using both classical taxonomical methods and molecular phylogenetic analysis. The phylogenetic reconstruction, based on four chloroplast DNA regions (matK, rbcL, trnLF, and psbAtrnH), was carried out employing maximum parsimony, maximum likelihood and Bayesian inference approaches. These analyses confirmed the systematic positioning of Artabotrys rubriflorus sp. nov. within the genus Artabotrys, while also establishing its distinctiveness from other species.
Methods
Chloroplast genomic DNA was extracted from collected silica-dried leaves (Narzary et al. 2015) using the Ezup Spin Column Super Plant Genomic DNA Extraction Kit (Sangon Biotech, Shanghai). Four chloroplast DNA regions (matK, rbcL, trnL-F and psbA-trnH) were subjected to PCR amplification. Primer information is given in Table 3. The PCR reaction system (25 µl) consisted of 15 µl of 2 Taq Master Mix buffer, 10 µl of ddH2O, 3 µl of the primer mixture (1.5 µl each of the positive and reverse primer), and 2 µl of the DNA template. The parameters for PCR procedure was 95 for 3 min, followed by 40 cycles of 95 for 30 s, appropriate annealing temperature for 30 s, 72 for 1 min, and 10 min for final extension at 72 , the annealing temperatures was 54 for matK and rbcL genes, 49 for trnL-F and psbA-trnH intergenic spacers. 5 µl of the products were tested using electrophoresis in 1% agarose gel at 150 V, 110 mA for 10 to 20 min. San Prep Column DNA Gel Extraction Kit (Sangon Biotech, Shanghai) was used for recovery and purification, then the purified products were sequenced by Sangon Biotech (Shanghai, China  https://www.sangon.com/).