Systematics Of Anaphalis From Pan-himalayas

Anaphalis DC.belongs to the tribe Gnaphalieae,family Compositae/Asteraceae.It is the largest genus in Asian Gnaphalieae tribe,containing about 110 species.PanHimalayas region,including Mount Himalaya and its neighboring regions,is the hotspot of plant taxonomy and phylogenetic studies as a natural geographic unit.This area is the diversity center of Anaphalis,with abundant Anaphalis species(60-70% in total).Anaphalis is one of the most problematic taxonomical group in Asteraceae,due to its large number of species,overlapping morphological characters and obscure species delimitation.This study aims to investigate documents and herbarium specimens of Anaphalis spp.to find out Anaphalis species and their distributions in this area.Achene surface micro-morphology and molecular systematics of Anaphalis were also studied to construct the relationships within Anaphalis,as well as its relationships with closely related genera.The results provide evidence for taxonomy study of Anaphalis and Gnaphalieae,as well as the plant diversity research of Pan-Himalayas region.1.Achene surface micro-morphology research of AnaphalisAchenes of 27 Anaphalis species were observed by electron microscope to investigate the achene surface characters.The shapes,size,cellular arrangement,cell shape,ornamentation of outer cell walls and epicuticular secretion of achenes were surveyed and compared.The results showed that micro-morphological characters of achene surface are distinct between species.According to the differences of achene surface ornamentation,Anaphalis can be divided into two groups: one group is with netted and rodlike bulge,including A.margaritacea and A.margaritacea var.Angustifolia from ser.Margaripes(DC.)Boiss.,A.busua from ser.Busuae Ling and all species of ser.Nepalenses Ling;the other groupe is with ligule bulge,including A.contortiformis from ser.Margaripes(DC.)Boiss.and all species from ser.Sinicae Ling.,ser.Suffruticosae Ling.,ser.Flavescentes Ling.,ser.Xylorhizae Ling.and ser.Pennosae Ling.The results did not support traditional classification of Anaphalis sections and series based on macro-morphological characters.There were overlapping micro-morphological characters between different series of Anaphalis.2.Molecular systematics of AnaphalisIn this study,three chloroplast DNA sequences,including psb A-trn H,trn L-trn F and rpl32-trn L intergenic spacer,and one nr DNA sequence,ITS,were sequenced from twenty-five Anaphalis species,combined with relevant species sequences from Genbank to construct phylogenetic trees and study the phylogenetic relationships within the Anaphalis and between related genera..The results indicated that: 1)Anaphalis is not monophyletic.There are two clades within Anaphalis are well recognized both in cp DNA and nr DNA trees.One clade involved most of Anaphalis species,and the other clade only included five Anaphalis species(A.nepalensis,A.margaritacea,A.triplinervis,A.porphyrolepis,A.virgata and one Gnaphalium specise G.adnata was included).Additionally,all Anapalis species used are nested with Helichrysum.2)In the phylogenetic tree based on ITS sequences,A.adnata,A.margaritacea,A.nepalensis,A.triplinervis,A.porphyrolepis and A.virgata are in one clade and supported well(Clade G,BSML=96%,PP=1).However,this result did not support the opinion of transfering A.adnata to Pseudognaphalium in Flora of China.3)Anaphalis is very close related to Helichrysum and Pseudognaphalium.4)There were inconsistence between the phylogenetic relationships within Anaphalis based on cp DNA data and nr DNA data.3.Taxonomy of Anaphalis in the Pan-Himalayan regionWe confirmed that there were 66 Anaphalis species and 21 varietas in PanHimalayas region by studying flora and plant list of Pan-Himalayas region,related websites,monograph,original literatures and herbarium specimens of Anaphalis.According to the herbarium specimens we have studied and calculated,the distribution of Anaphalis in this area was increased in detail.Key of Anaphalis in Pan-Himalayas region was made by analyzing and comparing the morphological characters.According to macro-morphology,micro-morphology and molecular systematics research,Anaphalis species can be divided into two groups: ?.Leaf base not decurrent group,including Ser.Margaripes(DC.)Boiss.Ser.Nepalenses Ling;II.Leaf base decurrent group,including Subgen.Gnaphaliops Ling,Ser.Busuae Ling,Ser.Virgatae Boriss.,Ser.Sinicae Ling,Ser.Oxyphyllae Ling,Ser.Suffruticosae Ling,Ser.Xylorhizae Ling,Ser.Pannosae Ling and Ser.Flavescentes Ling,was not consistent with the traditional classification.The leaf base not decurrent A.margaritacea,A.nepalensis,A.triplinervis and A.acutifolia phylogenetic relationship,in molecular systematics were clustered into one clade,and has netted and rodlike bulge;leaf base system this study involves the extension group relationship,in the molecular systematics of clustered into one,and with ligule bulge.