牛東玲, 朱　強(qiáng) , 王忠忠 , 石　晶 , 白明生 , 鄭學(xué)平

(1 寧夏大學(xué) 生命科學(xué)學(xué)院,銀川 750021；2 寧夏林業(yè)研究所,銀川 750000)

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地卷屬1中國(guó)新記錄種

牛東玲1, 朱強(qiáng)2, 王忠忠1, 石晶1, 白明生1, 鄭學(xué)平1

(1 寧夏大學(xué) 生命科學(xué)學(xué)院,銀川 750021；2 寧夏林業(yè)研究所,銀川 750000)

摘要:通過(guò)對(duì)地卷屬地衣形態(tài)和化學(xué)的研究，并結(jié)合核基因ITS序列的系統(tǒng)發(fā)育分析，報(bào)道了采自中國(guó)西北地區(qū)地卷屬的1個(gè)中國(guó)新記錄種——芽片地卷(新擬)。它的典型特征是沿著地衣體邊緣或地衣體上表面裂隙具有大量的薄片狀phyllidia，且常常覆有粉霜。地衣體上表面的邊緣具有白色絨毛，下表面具有叢生假根,而且假根在邊緣為白色，逐漸向中心變?yōu)樯钌Ｔ撗芯刻峁┝嗽摲N的詳細(xì)描述，并與近緣種進(jìn)行了細(xì)致的討論。

關(guān)鍵詞:中國(guó)西北; 地卷屬; ITS序列；新記錄種;

The genusPeltigerais cosmopolitan monophyletic genus with more than 91 species, which typically have muscicolous and terricolous foliose thallus[1-3]. Most ofPeltigeraspecies have a cyanobacterial photoboint that carries out nitrogen fixation and plays a major ecological role in nitrogen cycling of ecosystem. This genus is a common lichen group represented by 28 taxa in China[4-5], distributed in all geographical distributions of China. However, until now this genus is not studied thoroughly in China, in which the potential for finding new species is high. During our recent fieldwork in Liupanshan and Helanshan Mountains located in the northwest of China, we find one additional taxa (PeltigeramonticolaVitik.) belonging to this genus based on morphology and molecular characters.

1Materials and methods

1.1Morphological analysis

Specimens for this study were collected from Liupanshan and Helanshan Mountains, located in the northwest of China. The morphological descriptions were made from dry materials. 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 were 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)

1.2Chemical analysis

Chemical analyses were carried out using spot test[6]. These involve applying tiny amounts of a reagent to lichen and observing any resulting color change. Four reagents were used in these spot tests, K (10% aqueous potassium hydroxide), C (sodium hypochlorite solution), KC (10% K followed by C solution), P (5% alcoholic P-phenylenediamine).

1.3Molecular analysis

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

Table 1　Voucher information of Peltigera monticola Vitik. samples with GenBank accession number and specimens from GenBank ITS sequences obtained

*Specimen name followed by an asterisk represents outgroup sequence.

1.4Phylogenetic analysis

The entire ITS sequences of 6 samples examined and 14 representatives selected (Table 1) were aligned by Muscle implemented inMEGAversion 6[11]. The alignments were analyzed using neighbor joining (NJ), the software-based Kimura 2-parameter model was used, and gaps were excluded in the pairwise distance estimation, the support values were tested by 1 000 bootstrap replications.

2Results and discussion

2.1Phylogenetic analyses

Sequences of ITS alone were sufficient to identify all species from thePeltigeracaninaspecies complex. TheP.ponojensisgroup was well defined as a monophyletic clade (Fig.1), which has been identified before[12]. ITS sequences of representative collections showed that all studiedPeltigeraspecies from Liupanshan and Helanshan Mountains, belonged to the speciesP.monticolaclade, support for the monophyly of this taxon was strong (BS=78%). The specimens in different geographical region were in different lineages, the specimens from Liupanshan were in a lineage, those from Helanshan in another lineage. ‘P.scotteri’ is a nonmonophyletic putative species, and one of the specimens has been proved sister toP.monicola, so ‘P.scotteri’ is not validly published[13]. The specimen of ‘P.scotteri’ from Canade (Table 1) belonged to Helanshan lineage, which means this ‘P.scotteri’ should be treated as a member ofP.monticola. The specimens from Liupanshan were sister to the specimens from Poland with high support (BS=99%), the specimens from Helanshan and Switzerland were delimited as a monophyletic entity (BS=97%). Sequence data were also compared using Blast analysis of ITS1-2 regions deposited in NCBI. They were 100 % identical toPeltigeramonticola.

2.2The species

PeltigeramonticolaVitik., Acta Bot. Fennica 152: 64 (1994,Fig. 2)

Thallus: foliose, small to medium-sized, 3-9 cm in diam., adnate; lobes: flattened and elongate, 0.5-3 cm long and 1-3 mm wide, with up-turned, curled, with numerous flat phyllidia; upper surface: gray, partially brown when dry, tomentose marginally, dull marginally, smooth forwards central, sometimes pruinose on the surface; phyllidia: flat squamulose along margins and cracks, often pruinose; medulla: white with interwoven hyphae; photobiont: nostoc; lower surface: white margin with white raised veins, brown towards center; rhizines: tufted, pale near margins,darkened towards center; apothecia: not seen, said to be flat or saddle-shaped, up to 5 mm in diam.; pycnidia: not seen.

Support is indicated for branches by bootstrap frequencies exceeding 50% under the Neighbor JoiningFig. 1　Phylogenetic tree of P. monticola and selected species of P. ponojensis group based on ITS nrDNA

A. general habit; B. marginal phyllidia with pruinose; C. pruinose on the surface; D. tufted rhizinesFig.2 Peltigera monticola.

Chemistry: spot tests on the cortex and medulla K-, C-, KC-, P-.

Ecology and Distribution:P.monticolamainly grows among mosses over soil. Its world distribution includes North America, Europe and Asia[14]. At present this species is mainly recorded from Liupanshan which is located in Ningxia and Helanshan which is located in Ningxia and Inner Mongolia from the Northwest of China. Furthermore, it is mainly collected from forest zones in Nature Reserve of Ningxia and Inner Mongolia.

Notes: ITS results indicate that this taxon from the northwest of China is comprised of two geographically distinct lineages, but there is no obvious morphological difference between them, so two geographical populations ofP.monticolaare treated here as a taxon. The population from the northwest of China is sister to the specimens from Europe (Poland and Switzerland) with high support. We speculate it is due to their similar climatic conditions.P.monticolais identified by its laminal phyllidia along the margin or cracks, lobe ends with grey-whitish tomentose, tufted rhizines. The species is close toP.ponojenisbasing on GenBank Blast, but the latter is distinguished by short of phyllidia and pale unbranched to fibrous, discrete rhizines.

Specimens examined: China, Prov. Ningxia, Guyuan, Liupanshan, Qiuqianjia, 1 718 m, on moss, 2012, 12-0016; Liupanshan, Erlonghe, 2 039 m, on moss, 2012, 12-0017; Helanshan, Suyukou, 2 086 m, on moss, 2013,13-0005; 13-0004; Prov. Inner Mongolia: Helanshan, South Temple, 2 000 m, on moss, 2013, 13-0002; 13-0003. -(specimens are deposited in NXAC). France: Lorraine, La Madeleine, Fusboesch, 345 m, on soil, 2008, 07.27.02/8.- (specimens are deposited in Duke University Herbarium).

References:

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(編輯:潘新社)

文章編號(hào):1000-4025(2016)06-1245-05

doi:10.7606/j.issn.1000-4025.2016.06.1245

收稿日期:2016-03-10;修改稿收到日期:2016-05-11

基金項(xiàng)目:國(guó)家自然科學(xué)基金(31360040)

作者簡(jiǎn)介:牛東玲(1973- )，女，博士，教授，主要從事植物分類及植物化學(xué)研究。E-mail: niudl@nxu.edu.cn

中圖分類號(hào):Q949.34

文獻(xiàn)標(biāo)志碼:A

New Record of the Lichen GenusPeltigerafrom China

NIU Dongling1, ZHU Qiang2, WANG Zhongzhong1, SHI Jing1,BAI Mingsheng1, ZHENG Xueping1

(1 Life Science School, Ningxia University, Yinchuan 750021, China; 2. Ningxia Forest Research Institution, Yinchuan 750000, China)

Abstract:A new record species Peltigera monticola Vitik. was identified from the northwest of China, based on morphological and chemical analysis, and a phylgenetic analysis of nrDNA ITS sequence was also carried out. It was distinguished by its numerous laminal phyllidia, often pruinose, along the margin or cracks, marginal parts tomentose, tufted rhizomes with pale near margins and darkened towards center. Detailed description and discussion with allied species were provided.

Key words:northwest of China; Peltigera; ITS sequences; new record species