Comparative chloroplast genomics of three species of Bulbophyllum section Cirrhopetalum (Orchidaceae), with an emphasis on the description of a new species from Eastern Himalaya

Background Chloroplast (cp) genomes are useful and informative molecular markers used for species determination and phylogenetic analysis. Bulbophyllum is one of the most taxonomically complex taxa in Orchidaceae. However, the genome characteristics of Bulbophyllum are poorly understood. Methods Based on comparative morphological and genomic analysis, a new species Bulbophyllum pilopetalum from eastern Himalaya belonging to section Cirrhopetalum is described and illustrated. This study used chloroplast genomic sequences and ribosomal DNA (nrDNA) analysis to distinguish the new Bulbophyllum species and determine its phylogenetic position. An additional phylogenetic analysis was conducted using 74 coding sequences from 15 complete chloroplast genomes from the genus Bulbophyllum, as well as nrDNA sequences and two chloroplast DNA sequences from 33 Bulbophyllun species. Results The new species is morphologically similar to B. pingnanense, B. albociliatum, and B. brevipedunculatum in vegetative and floral morphology, but it can be distinguished by its ovate-triangle dorsal sepal without a marginal ciliate. The chloroplast genome of the new Bulbophyllum species is 151,148 bp in length, and includes a pair of inverted repeats (IRs) of 25,833 bp, a large single-copy region (LSC) of 86,138 bp, and a small single-copy region (SSC) of 13,300 bp. The chloroplast genome includes 108 unique genes encoding 75 proteins, 30 tRNAs, and four rRNAs. Compared with the cp genomes of its two most closely-related species, B. pingnanense and B. albociliatum, this chloroplast genome exhibited great interspecific divergence and contained several Indels that were specific to the new species. The plastid tree showed that B. pilopetalum is most closely-related to B. pingnanense. The phylogenetic tree based on combined nrDNA and chloroplast DNA sequences indicated that section Cirrhopetalum was monophyletic and B. pilopetalum was a member of this section. Discussion The taxonomic status of the new species is strongly supported by cp genome data. Our study highlights the importance of using the complete cp genome to identify species, elucidate the taxonomy, and reconstruct the phylogeny of plant groups with complicated taxonomic problems.

[1]  Shicheng Shao,et al.  Chloroplast genomic diversity in Bulbophyllum section Macrocaulia (Orchidaceae, Epidendroideae, Malaxideae): Insights into species divergence and adaptive evolution , 2021, Plant diversity.

[2]  E. Teoh Bulbophyllum Thouars. , 2021, Orchid Species from Himalaya and Southeast Asia Vol. 1 (A - E).

[3]  M. Chase,et al.  Comparative Plastid Genomics of Neotropical Bulbophyllum (Orchidaceae; Epidendroideae) , 2020, Frontiers in Plant Science.

[4]  R. Saunders,et al.  Molecular phylogenetics and floral evolution of the Cirrhopetalum alliance (Bulbophyllum, Orchidaceae): evolutionary transitions and phylogenetic signal variation. , 2019, Molecular phylogenetics and evolution.

[5]  HONG-BO Jiang,et al.  Bulbophyllum reflexipetalum (Orchidaceae, Epidendroideae, Malaxideae), a new species from Xizang, China , 2019, PhytoKeys.

[6]  Mei Jiang,et al.  CPGAVAS2, an integrated plastome sequence annotator and analyzer , 2019, Nucleic Acids Res..

[7]  H. Comes,et al.  Clade-age-dependent diversification under high species turnover shapes species richness disparities among tropical rainforest lineages of Bulbophyllum (Orchidaceae) , 2019, BMC Evolutionary Biology.

[8]  R. Bock,et al.  OrganellarGenomeDRAW (OGDRAW) version 1.3.1: expanded toolkit for the graphical visualization of organellar genomes , 2019, bioRxiv.

[9]  D. Albach,et al.  Extremely low levels of chloroplast genome sequence variability in Astelia pumila (Asteliaceae, Asparagales) , 2019, PeerJ.

[10]  Zhiduan Chen,et al.  Combining complete chloroplast genome sequences with target loci data and morphology to resolve species limits in Triplostegia (Caprifoliaceae). , 2018, Molecular phylogenetics and evolution.

[11]  Jiang-Feng Liu,et al.  Bulbophyllum yongtaiense (Orchidaceae, Epidendroideae, Dendrobiinae), a new species from Fujian, China: evidence from morphological and molecular analyses. , 2018 .

[12]  Wen-Bin Yu,et al.  GetOrganelle: a fast and versatile toolkit for accurate de novo assembly of organelle genomes , 2018, Genome Biology.

[13]  Jiang-Feng Liu,et al.  Bulbophyllum yunxiaoense sp. nov. (Orchidaceae: Epidendroideae: Malaxideae) from Fujian, China: Morphological and molecular analyses , 2017 .

[14]  Axel Fischer,et al.  GeSeq – versatile and accurate annotation of organelle genomes , 2017, Nucleic Acids Res..

[15]  Y. Yarom,et al.  Replacing Sanger with Next Generation Sequencing to improve coverage and quality of reference DNA barcodes for plants , 2017, Scientific Reports.

[16]  Jiang-Feng Liu,et al.  Bulbophyllum pingnanense (Orchidaceae, Epidendroideae, Dendrobiinae), a new species from Fujian, China , 2016, PhytoKeys.

[17]  M. Chase,et al.  An updated classification of Orchidaceae , 2015 .

[18]  Shih‐Wen Chung,et al.  Two New Bulbophyllum (Orchidaceae) Species in Taiwan: B. brevipedunculatum and B. ciliisepalum , 2008 .

[19]  L. Garay,et al.  The genus Cirrhopetalum and the genera of the Bulbophyllum alliance , 1994 .

[20]  Du Petit-Thouars,et al.  Histoire particulière des plantes Orchidées recueillies sur les trois îles australes d'Afrique, de France, de Bourbon et de Madagascar / , 1822 .