寧夏地衣一新記錄屬
——金黃衣屬及其新記錄種

牛東玲，王繼飛，田曉燕

（1寧夏大學(xué)，銀川750021；2寧夏賀蘭山國(guó)家級(jí)自然保護(hù)區(qū)管理局，銀川750000）

寧夏地衣一新記錄屬
——金黃衣屬及其新記錄種

牛東玲1，王繼飛2，田曉燕1

（1寧夏大學(xué)，銀川750021；2寧夏賀蘭山國(guó)家級(jí)自然保護(hù)區(qū)管理局，銀川750000）

報(bào)道了寧夏地衣的1個(gè)新記錄屬——金黃衣屬及其新記錄種——戈壁微孢衣。利用形態(tài)學(xué)和分子生物學(xué)的手段對(duì)該種進(jìn)行了分類學(xué)鑒定，并對(duì)該種的形態(tài)、化學(xué)及生態(tài)分布等方面進(jìn)行了詳細(xì)記述。調(diào)查發(fā)現(xiàn)該種對(duì)人類的干擾活動(dòng)非常敏感。

荒漠地衣；寧夏植被；戈壁微孢衣

The lichen genus Pleopsidium was first described by Koerber in 1855［1］.Later it was proposed as a section of Acarospora［2］，and then was raised to generic rank by Hafellner［3］.Genus Pleopsidiumwas mainly distributed in Eastern A－sia，including Mongolia and China［3－4］.In China，it was found in Gansu，Qinghai，Hebei，Shanxi，Yunnan，Sichuan and Tibet［4－5］.Pleopsidium mainly differs from Acarosporain structure of ascus and upper cortex of thallus［3］.Pleopsidiumis a small genus with four species found worldwide：Pleopsidium flavum，P.chlorophanum，P.discurrens and P.gobiense.During collecting trips of desert li－chens along the Eastern foot of Helan Mountain in Northwest China，P.gobiense was identified for the first time to exist in Ningxia.Earlier studies on Ningxia flora contain no reference to this genus.The present discovery，therefore，is a new generic record for Ningxia flora.

1 Material and Methods

1.1 Morphological analysis

Specimens for this study were collected from the Eastern foot of Helan Mountain，Ningxia.The morphological descriptions are made from dry materials.A freezing microtome Leica CM1950（Leicainstrument Co.，Germany）was used for preparing anatomical sections.Morphological and anatomical characters were observed and photographed using a XTS20（Fukai Science and Technology Development Co.，China）and Leica DM2500（Leica instrument Co.，Germany）.The voucher specimens for this study are kept in the herbarium of College of Life Science of Ningxia University（NXAC，Yinchuan，Ningxia，China）.Besides our own collection we examined the specimens of the same species deposited in William Louis ＆Chicita F.Culbertson Lichen Herbarium ＆Library（Duke University，Durham，NC，USA）

1.2 Chemical analysis

Chemical analyses were carried out using spot test［6］.

1.3 Molecular analysis

DNA was obtained from fresh sample（10－0199），extraction followed a modification of DOYLE and DOYLE’s（1987）［7］method［8］.rRNA internal transcribed spacers（ITS1－2）were amplified via polymerase chain reaction（PCR）using following primers：ITS1F［9］and ITS4［10］.PCR mixtures（25μL）consisted of：25μg BSA，1UTaq DNA polymerase，dNTPs（0.2mmol／L），primers（0.5μmol／L each），and PCR buffer，adjusted with H2O to 25μL.The following thermal cycler profile was employed for PCR reactions：an initial denaturation of 95℃for 5min，follow by 35cycles of 95℃for 45s，52℃for 90s，and 72℃for 90s，with a final extension of 72℃for 10min.Amplification products were cleaned with ExoAP and sequenced using Big Dye chemistry with an ABI 3730automated sequencer（PE Applied Biosystems，USA）.Sequences were assembled using Sequencher 4.2（Gene Codes，USA）.The ITS sequences were subjected to BLAST searches to verify their identity.

2 Results

Pleopsidium gobiense（H.Magn.）Hafellner（1993）（Plate I，F(xiàn)ig.A－Fig.G）

Bas.：Acarospora gobiensis Magnusson，Kungl.Svensk.Vetensk.Akad.Handl.7／4：98（1929）

Thallus：crustose，distinctly radiating margin with lobes 1－2mm long，up to 2cm or more across；areoles：irregularly shaped，convex，0.5－1.5mm across，mostly sterile，mostly densely contiguous；upper surface：bright yellowish green，smooth and becoming rugose toward center；upper cortex：78.8－115.4μm thick，half cortex yellow，composed of short－celled prosoplectenchyma；algal layer：46.2－192.3μm thick，irregularly interrupted by hyphal bundles；medulla：thickness varying，about 115.4－500.0μm，intricate，prosoplectenchymatous with granules，without a lower surface；photoboint：green，unicellular；apothecia：lecanorine，0.5－3mm in diam.；disk：yellowish green，darker than thallus，initially immersed in the areolate，becoming plane to convex，sometimes corrugated and having cracks in older apothecia，margin sometimes excluded；true exciple：hyaline，76.9－115.4 μm thick；epihymenium yellow 9.6－13.5μm thick；hymenium：hyaline，57.7－67.3μm high，paraphyse septate，tips no swollen；subhymenium：hyaline，38.5－96.2μm；hypothecium：hyaline，19.2－28.8μm thick，paraphyse septate；asci：clavate，（9.6－13.5）×（19.2－28.9）μm，with well－developed tholus，K／I＋blue，many per ascus；ascospores：unicellular，small，hyaline，ellipsoid，（0.96－1.9）×（1.9－3.9）μm；pycnidia：deeply immersed with dark ostioles，many pycnidia clustered together；conidia：（0.5－1.9）×（1.9－2.1）μm.

Chemistry：spot tests on the cortex and medulla K－，C－，KC－，P－，cortex and epihymenium with bright yellow pigment（rhizocarpic acid），some cells in the cortex full of bright yellow crystals.

Ecology and Distribution：In China this species is the first time found on the surfaces of biotite plagiogranite which scatter along the top of diluvial of Eastern foot of Helan Mountain.It is also distributed in Mongolia［3］.This species was mainly recorded from the less disturbed locations，and sensitive to human disturbance.

Sequence data were compared using BLAST analysis of ITS1－2regions deposited in NCBI；they are 100%identical to Pleopsidium gobiense.Sequence data was submitted to GenBank（ACC＃KP641170）.

3 Discussion

This is the first report of Pleopsidium gobiense from Ningxia.Four species in genus Pleopsidiumare very similar in appearance；P.gobiense differs from other three members of the genus by its thallus size，apothecia，chemistry and ecological distribution.Apothecia of P.gobiense vary from immersed，becoming plane to convex，with or without margin，this character is shared with P.flavum and P.chlorophanum.P.gobiense has a varying size from about 2cm to more than 10cm corresponding to the degree of interference.In contrast，the thallus of P.chlorophanumis less than 2cm across；P.flavumhas a bigger thallus size，up to 6cm or more［11］.P.gobiense was found on the surfaces of biotite plagiogranite scattered on the diluvia，different fromP.chlorophanumand P.flavum which occurred on overhanging stone.Up to now，P.gobiense is only found in East Asia（Mongolia and China）.P.chlorophanumis distributed in Antarctica，Europe，South and North America at high elevations.P.flavumhas a broader ecological amplitude，and it was found in Europe，North America，Africa and Asia［11］.P.disturrens was mainly distributed in China［5］.According to Hafellner，P.gobiense contains rhizocarpic acid［3］，but Pleopsid－ium disturrens，P.flavumand P.chlorophanum additionally contains other acids：acaranoic，acarenoic，roccellic or lichesterinic acids［11］.

The locality where P.gobiense was found scatters along the diluvial of the eastern foot of Helan Mountain.In the disturbed area of human activities including grazing and industrial exploitation，P.gobienseis very rare，and its thallus diameter does not exceed 2cm，individual thallus has only several areoles.However in the protected areas，P.gobienseis found frequently and its thallus diameter is bigger，more than 10cm.It is obvious that this species is sensitive to human disturbance.Further study is required for better understanding of P.gobiense status in nature.

Specimens examined：China：Prov.Ningxia：Yinchuan City，the Eastern foot of Helan Mountain，Maliankou，1 366m，2010，10－0199；1 694.8 m，2014，14－01－413；Suyukou，1 400m，2010，10－0303；1 398.3m，2014，14－01－0145；1 365.9m，2014，14－03－0183；1 342.1m，2014，14－06－0227；Dakouzi，1 400m，2010，10－0287；Baisikou，1 323m，2010，10－0226－（specimens are deposited in NXAC）.Mongolia：Gobi Altai region，1 500m，1973，No.431－（specimens are deposited in Duke University Herbarium）.

Acknowledgements

I thank Professor Francois Lutzoni in Duke Univesity for offering the valuable resources and lab conditions，I am also grateful of Dr.Andrii Gryganskyi for translation of relevant German literature，review the manuscript and provide me with inspiring advice.This work is supported by the National Natural Science Foundation of China for the study of desert lichens in Northwest China（31360040）.

［1］ KOERBER G W.Systema Lichenum Germaniae［M］.Breslau：BiblioBazaar Publisher.2009.

［2］ MAGNUSSON A H.A monograph of the Scandinavian species of the genus Acarospora［J］.Goteborgs Kungl.Vetensk.Vitt.Handl.，1924，4.Fol.，28（2）：1－149.

［3］ HAFFLLNER J.Acarospora und Pleopsidium－zwei lichenisierte Ascomycetengattungen（Lecanorales）mit zahlreichen Konvergenzen［J］.Nova Hedwigia，1993，56（3／4）：281－305.

［4］ WEI J C.An Enumeration of Lichens in China［M］.Beijing：International Academic Publishers，1991：19.

［5］ OBERMAYER W.Pleopsidium discurrens（Zahlbr.）Obermayer comb.nova，newly discovered in southern Tibet（Lichenological results of the Sino－German Joint Expedition to southeastern and eastern Tibet 1994.II）［J］.Annales Botanici Fennici，1996，33：231－236.

［6］ BRODO I M，SHARNOFF S D，SHARNOFF S.Lichens of North America［M］.New Haven and London：Yale University Press，2001：103－108.

［7］ DOYLE J J ＆DOYLE J L.A rapid DNA isolation procedure for small quantities of fresh leaf tissue［J］.Phytochem.Bull..1987，19：11－15.

［8］ GOFFINTE B，BAYER R J，VITT D H.Circumscription and phylogeny of the Orthotrichales（Musci）based on rbc Lsequences［J］.Am J Bot.1998，85：1 324－1 337.

［9］ GARDES M，BRUNS T D.ITS primers with enhanced specificity for basidiomycetes—application to the identification of mycorrhizae and rusts［J］.Mol.Ecol.，1993，2：113－118.

［10］ WHITE T，BRUNS T D，LEE S，TAYLOR J W.Amplification and direct sequencing of fungal ribosomal RNA genes for phylogenetics［M］／／INNIS M A，GELFAND D H，SNINSKY J J，WHITE T J.（ed.）：PCR Protocols：aguide to methods and applications.New York：Academic Press Inc，1990：315－322.

［11］ NASH III T H，GRIES C，BUNGARTZ F.Lichen Flora of the Greater Sonoran Desert Region.Volume III［M］.Arizona：Arizona State University Press，2007：274－276.

（編輯：潘新社）

PleopsidiumKoerber——a New Record Genus for Ningxia with Description of New Record Species

NIU Dongling1，WANG Jifei2，TIAN Xiaoyan1
（1Ningxia University，Yinchuan 750021，China；2Helan Mountain National Nature Reserve Administration，Yinchuan 750000，China）

The genus Pleopsidiumis new to the flora of Ningxia.A new record species Pleopsidium gobiense was identified and characterized using morphological and molecular methodology，described and illustrated based on these data.P.gobienseis sensitive to human disturbance.

desert lichens；Ningxia flora；Pleopsidium gobiense

Q949.34

A

1000－4025（2015）08－1697－04

10.7606／j.issn.1000－4025.2015.08.1697

2015－01－07；修改稿收到日期：2015－05－12

國(guó)家自然科學(xué)基金（31360040）

牛東玲（1973－），博士，副教授，主要從事植物分類學(xué)及植物化學(xué)研究。E－mail：niudl＠nxu.edu.cn