Chloroplast genomic diversity in Bulbophyllum section Macrocaulia(Orchidaceae,Epidendroideae,Malaxideae):Insights into species divergence and adaptive evolution

Bulbophyllum is the largest genus in Orchidaceae with a pan tropical distribution.Due to highly significant diversifications,it is considered to be one of the most taxonomically and phylogenetically complex taxa.The diversification pattern and evolutionary adaptation of chloroplast genomes are poorly understood in this species-rich genus,and suitable molecular markers are necessary for species determination and phylogenetic analysis.A natural Asian section Macrocaulia was selected to estimate the interspecific divergence of chloroplast genomes in this study.Here,we sequenced the complete chloroplast genome of four Bulbophyllum species,including three species from section Macrocaulia.The four chloroplast genomes had a typical quadripartite structure with a genome size ranged from 156,182 to 158,524 bp.The chloroplast genomes included 113 unique genes encoding 79 proteins,30 tRNAs and 4 rRNAs.Comparison of the four chloroplast genomes showed that the three species from section Macrocaulia had similar structure and gene contents,and shared a number of indels,which mainly contribute to its monophyly.In addition,interspecific divergence level was also great.Several exclusive indels and polymorphism SSR loci might be used for taxonomical identification and determining interspecific polymorphisms.A total of 20 intergenic regions and three coding genes of the most variable hotspot regions were proposed as candidate effective molecular markers for future phylogenetic relationships at different taxonomical levels and species divergence in Bulbophyllum.All of chloroplast genes in four Bulbophyllum species were under purifying selection,while 13 sites within six genes exhibited sitespecific selection.A whole chloroplast genome phylogenetic analysis based on Maximum Likelihood,Bayesian and Parsimony methods all supported the monophyly of section Macrocaulia and the genus of Bulbophyllum.Our findings provide valuable molecular markers to use in accurately identifying species,clarifying taxonomy,and resolving the phylogeny and evolution of the genus Bulbophyllum.The molecular markers developed in this study will also contribute to further research of conservation of Bulbophyllum species.

Plastome characteristics of Cannabaceae

Cannabaceae is an economically important family that includes ten genera and ca. 117 accepted species.To explore the structure and size variation of their plastomes, we sequenced ten plastomes representing all ten genera of Cannabaceae. Each plastome possessed the typical angiosperm quadripartite structure and contained a total of 128 genes. The Inverted Repeat(IR) regions in five plastomes had experienced small expansions(330 e983 bp) into the Large Single-Copy(LSC) region. The plastome of Chaetachme aristata has experienced a 942-bp IR contraction and lost rpl22 and rps19 in its IRs. The substitution rates of rps19 and rpl22 decreased after they shifted from the LSC to IR. A 270-bp inversion was detected in the Parasponia rugosa plastome, which might have been mediated by 18-bp inverted repeats. Repeat sequences, simple sequence repeats, and nucleotide substitution rates varied among these plastomes.Molecular markers with more than 13% variable sites and 5% parsimony-informative sites were identified, which may be useful for further phylogenetic analysis and species identification. Our results show strong support for a sister relationship between Gironniera and Lozanell(BS ? 100). Celtis, CannabisHumulus, Chaetachme-Pteroceltis, and Trema-Parasponia formed a strongly supported clade, and their relationships were well resolved with strong support(BS ? 100). The availability of these ten plastomes provides valuable genetic information for accurately identifying species, clarifying taxonomy and reconstructing the intergeneric phylogeny of Cannabaceae.

Aphis （Hemiptera： Aphididae） species groups found in the Midwestern United States and their contribution to the phylogenetic knowledge of the genus

A phylogeny of the genus Aphis Linnaeus, 1 758 was built primarily from specimens collected in the Midwest of the United States. A data matrix was constructedwith 68 species and 41 morphological characters with respective character states of alate and apterous viviparous females. Dendrogram topologies of analyses performed usingUPGMA （Unweighted Pair Group Method with Arithmetic Mean）, Maximum Parsimony and Bayesian analysis of Cytochrome Oxidase I, Elongation Factor 1-α and primary endosymbiont Buchnera aphidicola 16S sequences were not congruent. Bayesian analysis strongly supported most terminal nodes of the phylogenetic trees. The phylogeny wasstrongly supported by EFI-α, and analysis of COl and EFI-α molecular data combined with morphological characters. It was not supported by single analysis of COI or Buch-hera aphidicola 16S. Results from the Bayesian phylogeny show 4 main species groups： asclepiadis,fabae, gossypii, and middletonii. Results place Aphis and species of the generaProtaphis Bōrner, 1952, Toxoptera Koch, 1856 and Xerobion Nevsky, 1928 in a monophyletic clade. Morphological characters support this monophyly as well. The phylogenyshows that the monophyletic clade of the North American middletonii species group belong to the genus Protaphis： P. debilicornis （Gillette ＆ Palmer, 1929）, comb. nov., P. echinaceae（Lagos and Voegtlin, 2009）, comb. nov., and P. middletonii （Thomas, 1879）. The genus Toxoptera should be considered a subgenus of Aphis （stat. nov.）. The analysis also indicatesthat the current genus Iowana Frison, 1954 should be considered a subgenus of Aphis （stat. nov.）.