Causonissessilifolia (Vitaceae), a new species from Thailand

Abstract A new species, Causonissessilifolia, from Thailand is described, based on morphological and phylogenetical methods. A full description, conservation assessment, a key, images and phylogenetic tree are provided. Diagnostic characters for this species are sessile leaves that are sometimes opposite and inflorescence insertion interfoliar.

Causonis comprises about 16 species and four varieties and is found in tropical, subtropical and temperate Asia to Australia including the Pacific Islands (Parmar et al. 2021). Trias-Blasi and Parnell (2020) reported two Causonis species in Thailand, but the specimen from Nakhon Sawan in the northern floristic region of Thailand, was found to be morphologically distinct from other species reported from the region.
In autumn 2019, an expedition was conducted in a 5-hectare area adjacent to Kriangkrai River in the Nakhon Sawan Province (Fig. 1), because of concerns relating to high levels of deforestation in the lowland floodplain forests in Thailand. Additionally, very few specimens had been collected from this habitat. Generally, these areas are flooded every year during September-November when the water increases by 1-3 metres. This species was found in an open area along the riverbank.

Methods
This study is based on the material collected in October 2019 in Nakhon Sawan. Morphological characters were studied using a hand lens (30-60× magnification) and stereomicroscope and documented by photography. Collected specimens were thoroughly compared with protologues and types of all Causonis species occurring in Thailand and neighbouring regions. Additionally, herbarium material of Causonis, deposited in K, was studied (herbarium codes according to Thiers 2021). Herbarium vouchers for this study are deposited in BKF. The description follows the style and level of details outlined in Trias-Blasi and Parnell (2020), while the general terminology is based on Beentje (2016). The conservation assessment is based on the most recent version of the guidelines of IUCN Standards and Petitions Subcommittee (IUCN 2012).
Genomic DNA for the new taxon was extracted from silica-gel-dried leaf material using DNeasy Plant Mini Kit Qiagen (Qiagen, Hilden, Germany) following manufacturers' protocols. All other sequences for different taxa were downloaded from GenBank. The DNAs were amplified for four chloroplast loci (atpB-rbcL, trnC-petN, trnH-psbA and trnL-F) following the protocols in Lu et al. (2018). The PCR products were purified and examined on a 1% agarose gel before being sent to Majorbio Company in Beijing, China, for sequencing on a Roche 454 sequencer with the same PCR amplification primers using a standard GS FLX Titanium sequencing kit XL+ (454 Life Sciences, Branford, CT, USA). The voucher specimens and the sequences' Gen-Bank accession numbers are provided in Table 1.
Geneious 8.1.9 was used to assemble forward and reverse sequences (Kearse et al. 2012). Geneious was also used to edit contiguous sequences and check chromatograms for base validation. Following that, sequences were aligned using MAFFT 1.3.1 (Katoh et al. 2002) and then manually adjusted in Geneious. On the CIPRES Science Gateway (Miller et al. 2010), phylogenetic analyses were first performed for individ-ual DNA loci using the Maximum Likelihood (ML) approach in RAxML-HPC2 on XSEDE (8.2.12) using the GTR + G model with 1000 bootstrap replicates (Stamatakis 2014). Single tree analyses did not detect well-supported topological conflicts amongst individual DNA loci (i.e. ML BS < 70%; Hillis and Bull 1993). As a result, for further phylogenetic analyses, the four chloroplast DNA loci were concatenated. For the four chloroplast (4cp) dataset, partitioned ML and Bayesian Inference (BI) analyses were performed and the best fitting models for individual data partitions were selected using PhyML 3.0 (Guindon et al. 2010) with the Akaike Information Criterion (AIC). The nucleotide substitution model GTR + G was found to be the most suitable for trnC-petN and trnL-F and HKY85 + G for atpB-rbcL and trnH-psbA. MrBayes 3.2.6 was used to conduct Bayesian analysis on the CIPRES Science Gateway (Ronquist et al. 2012). For a total of 10,000,000 generations, four Markov Chain Monte Carlo analyses were conducted, with one tree sampled every 1,000 generations. The standard deviation between the split frequencies was found to be less than 0.01, indicating that enough generations had been completed. Following the burn-in of the first 25% of trees, the remaining trees were used to determine a 50% majority-rule consensus tree and posterior probabilities (PP). The trees obtained from ML and BI were analysed for topological conflicts through FigTree v.1.4.4 (Rambaut 2018).
Etymology. The specific epithet "sessilifolia" refers to the sessile leaves of the taxon. Distribution and habitat. Thailand (Northern floristic region, Nakhon Sawan). Lowland floodplain forest, along the bank of canal in open areas; 30 m alt. (Figs 1, 3).
Conservation status. This species is only known from the type locality and, therefore, has an Area of Occupancy (AOO) and Extent of Occupancy (EOO) of 4 km 2 . This suggests that it might be Critically Endangered as the AOO is less than 10 km 2 and is only found in one location. The species has been found to grow outside any protected areas and in an abandoned area next to a canal. This could mean the species is more vulnerable than others as it is unprotected. Additionally, all the surrounding areas are used for agriculture and, therefore, it is likely this forested area might also be transformed for this use. Due to this threat, the restricted AOO and number of locations, we think that this taxon could be driven to being Critically Endangered or Extinct in a very short time; therefore, we assess the taxon as VUD2 (IUCN 2012).