贵州麻阳河自然保护区7种野生红菇的形态与分子鉴定

2016-02-20 06:58谯利军文庭池康冀川吴兴亮
山地农业生物学报 2016年6期
关键词:红菇标本研究

谯利军,文庭池,康冀川*,康 超,吴兴亮

(1.贵州大学 西南药用生物资源教育部工程研究中心,贵州 贵阳 550025;2.贵州科学院,贵州 贵阳 570228)

贵州麻阳河自然保护区7种野生红菇的形态与分子鉴定

谯利军1,文庭池1,康冀川1*,康 超2,吴兴亮2

(1.贵州大学 西南药用生物资源教育部工程研究中心,贵州 贵阳 550025;2.贵州科学院,贵州 贵阳 570228)

为了解麻阳河自然保护区野生红菇真菌资源状况,从保护区采集到7种野生红菇,基于其形态特征结合ITS rDNA序列与GenBank中相关的红菇属的同类序列构建的分子系统发育树,将其鉴定为厌味红菇Russulanauseosa、美红菇Russulapuellaris、鳞盖红菇Russulalepida、菱红菇Russulavesca、以及变绿红菇Russulavirescens、Russulafaustiana、粉柄红菇Russulafarinipes。

野生红菇;形态分类;分子鉴定;ITS rDNA

红菇属RussulaPers.(1796)在分类学上隶属于担子菌门Basidiomycota、担子菌纲Basidiomycetes、红菇目Russulales Kreisel ex P.M.Kirk ,P.F.Cannon & J.C.David、红菇科Russulaceae Lotsy,在真菌系统学研究中具有相当重要意义,也是系统发育中颇为重要的类群[1-2]。目前国际上已经发表的红菇名称有2600多个(www.indexfungorum.org,2016),约750多种,但因为红菇属真菌形态差异大,导致有许多同种异名或异名同种的现象发生,修改订正后,报道的红菇属真菌达300多种,而我国记载的就有90余种,其中可食用的有60多种[3]。宋斌等[4]通过收集并整理文献,汇总了中国的红菇属RussulaPers.真菌,包括179个名称记录。此外,调查表明,红菇属真菌广泛分布于我国的30个省区,在已报道的红菇属名称记录中,有效名称有159个,其中包括148个种、9个变种和2个变型。张鑫等[5]以显微生态学为基础,报道了亚洲新纪录种蜡味红菇;付立忠等[6]利用分子系统学鉴定方法,对采自浙江的四个红菇科菌株进行测序分析,以鉴定其亲缘关系;Das等[7]基于ITS基因序列构建的系统发育树,结合宏观和微观形态鉴定,报道了RussulashingbaensisK. Das & S.L. Mill.和RussulaThindiiK. Das & S.L. Mill.,它们是两个采自印度的红菇属新种。本研究通过形态学和ITS rDNA序列相结合的方法,对采自贵州省麻阳河自然保护区的七种红菇真菌进行了分类鉴定。

1 材料与方法

1.1 材料

本次试验的大型真菌实体标本,均于2013年10月2~5日期间在麻阳河自然保护区采集,并保存于贵州大学贵州省生化工程中心标本室。

1.2 试剂及仪器

真菌基因组提取试剂盒(Fungal gDNA Kit)(BIOMIGA公司),PCR扩增所用试剂[DNA Marker(广州东盛生物科技有限公司),50×TAE溶液(Solarbio),2×PCR Master Mix、ITS 1、ITS 4、ddH2O(上海前尘生物科技有限公司)。

数码相机(Nikon),显微镜(OLYMPUS BX41TF),凝胶成像系统以及PCR仪(Bio-Rad公司)等。

1.3 红菇的形态鉴定

根据记录本的记录及照片对标本的宏观形态进行整理、描述,描述的内容包括研究标本的生境、习性;菌盖的大小、形状、颜色;菌柄的长、直径、形状等;菌褶生长情况等。颜色的记录主要根据野外记录情况及拍摄的彩色照片,需参考相关的工具书,并结合国际色彩标准名称与色值的电子版描述子实体颜色[8-9]。

1.4 红菇的分子鉴定

1.4.1 DNA提取

将红菇研磨成粉末状,取约10~50 mg,采用真菌基因组提取试剂盒提取红菇子实体的DNA,方法按照说明书进行。

1.4.2 PCR扩增和测序

以适量基因组DNA为模板,进行PCR扩增,扩增引物为ITS 1:5’-3’(TCC GTA GGT GAA CCT GCG G)和ITS 4:5’-3’(TCC TCC GCT TAT TGA TAT GC)[10]。PCR扩增的反应体系采用25 μL体系,包括有10×扩增缓冲液(含MgCl2)2.5μL,200μm dNTP 2.5 μL,5 μm的引物各1 μL,Taq DNA 聚合酶0.3 μL(2.5 u/μL),DNA 模板1.0~2 μL,用ddH2O补至25μL。PCR 扩增反应在PCR仪上进行。扩增程序具体参数如下:95℃预变性4 min,然后:94℃变性30 s,50℃(有时55℃)退火40 s,72℃延伸90 s,持续35个循环,最后再72℃延伸10 min,于4℃保存。经凝胶电泳检测后,适合测序的PCR产物送上海立菲生物技术有限公司测序。

1.4.2 序列比对及分子系统学分析

使用NCBI中Blast在线检索,下载与研究样品相近的ITS rDNA参考序列菌株信息(见表1),采用MEGA5.05[11]分别比对(alignment)测得的各个样本的基因片段与在GenBank比对后下载的相关真菌序列。再用MEGA5.05手工校正来实现排序匹配的最优化。比对后的序列采用PAUP* 4.0软件以最大简约法(Maximum parsimony, MP)进行分析,以启发式搜索法(Heuristic Search)获得系统发育树。其中,启发式搜索采用二等分再连接法(Tree-Bisection-Reconnection, TBR)作为获取聚类树的方法。在系统发育分析中碱基序列的空缺(Gaps)作为碱基缺失处理;把所有的碱基状态处理为无序,并给以相同的权重(转换和颠换的比率为1∶1),即不加权(Equal weight)处理。经1000次重复计算自展检验的值(bootstrap value)以评价系统发育树拓扑结构的可靠性,并标记在树的分枝上。最后通过Treeview V32软件对树进行编辑处理后导出为图片用PPT编辑(Page 2013)。

2 结果与分析

2.1 形态学鉴定

2.1.1 厌味红菇

Russulanauseosa(Pers.) Fr.

菌盖:30~31×25~28 mm,碟状或浅杯状,近圆形,中部略凹陷,印度红色,由中部到边缘逐渐变浅,边缘浅粉色,表面有黏性;菌褶:弯曲、等长、浅黄色,靠近菌柄处有少量分叉;菌柄:长40 mm,直径10 mm,空心,圆柱形,乳白色,基部杵状,肉质薄而易碎(见图1.a),本实验研究标本:MYH201310045。

生长环境:生于针叶林地上。

该种主要分布于吉林、云南、四川等省。

图1 7种野生红菇形态图Fig.1 7 species of wild Russula(a.厌味红菇(MYH201310045);b,c.美红菇(XJ2013100413);d.鳞盖红菇(XJ20131004125);e,f.菱红菇(XJ2013100418);g,h.绿菇(XJ2013100520);i,j.Russula faustiana(XJ2013100408);k,l.粉柄红菇(XJ2013100498))

2.1.2 美红菇

RussulapuellarisFr.

菌盖:45 mm×45 mm,中部略下凹,后平展,浅粉红色,有粘性;菌褶:较密、等长,白色,干后乳黄色;菌柄:长7 cm,直径:7 mm,圆柱形,直生,实心至中空,无附属物,海绵质,与菌褶同色,基部杵状(见图1.b、图1.c),本实验研究标本:XJ2013100413。

生长环境:生于混交林地上。

该种主要分布于中国广东、海南、四川、贵州、辽宁、吉林、江苏、江西、云南、西藏、河南、湖南、陕西,欧洲、北美洲等地。

2.1.3 鳞盖红菇

RussulalepidaFr.

菌盖:42~51 mm×37~45 mm,扁半球形,猩红色,无粘性。菌肉白色,质厚,味道鲜美;菌褶:密、等长,白色;菌柄:长33~45 mm,直径8~11 mm,圆柱形,实心,白色略带紫红色幼时紧密,老后松软(见图1.d),本实验研究标本:XJ20131004125。

生长环境:生于混交林地上。

该种在亚洲、欧洲和北美洲均有分布,均有分布我国湖北、湖南、广东、河北、辽宁、吉林、黑龙江、四川、贵州、云南、江苏、福建、江西、河南、广西、西藏 、陕西、甘肃等省。

2.1.4 菱红菇

RussulavescaFr.

菌盖:62 mm×71 mm,平展,有细微皱纹或近光滑,浅棕色,边缘有皱褶;菌褶:密、有分叉,延生,白色;菌柄:长50 mm,直径11 mm,圆柱形,内海绵质,靠近菌褶部分与菌褶同色,往下变黄或褐色,基部杵状。气味有淡淡的甜味(见图1.e、图1.f),本实验研究标本:XJ2013100418。

生长环境:生于混交林地上。

该种在亚洲、欧洲和北美洲均有分布,均有分布我国四川、贵州、云南、辽宁、西藏、新疆、江苏、福建、江西、湖南、广东、广西、海南,以及香港、台湾等地。

2.1.5 绿菇

Russulavirescens(Schaeff.) Fr.

菌盖:74 mm×80 mm,质地坚实,绿色,初呈球形,长至后来呈扁半球形,成熟时中央稍下凹,浅绿色至灰绿色,表皮往往斑状龟裂,形成卷毛或小碎片,老时边缘有条纹。菌肉白色,味道温和,无异味。菌褶:较密、等长,直生,具横脉,象牙白;菌柄:长45 mm,直径16 mm,圆柱形,短而粗,乳白色,基部圆头状,中实或内部松软(见图1.g、图1.h),本实验研究标本:XJ2013100520。

生长环境:生于混交林地上。

该种在亚洲、欧洲和北美洲均有分布,分布于我国的西藏、陕西、甘肃、内蒙古、辽宁、吉林、黑龙江、四川、贵州、云南、江苏、浙江、安徽、江西、山东、河南、湖北、湖南、广东、海南,以及香港、台湾等地。

2.1.6RussulafaustianaSarnari

菌盖:38 mm×40 mm,中间凹陷扁平,外有菌褶向上翻;菌褶:较密、不等长,贴生,有时分叉,白色;菌柄:长38 mm,直径12 mm,圆柱形,乳白色,笔直,基部杵状(见图1.i、图1.j),本实验研究标本:XJ2013100408。

生长环境:生于混交林地上。

该种在挪威有记录,而未在中国发现有记录。

2.1.7 粉柄红菇

RussulafarinipesRomell

菌盖:18 ~55 mm×14~46 mm,扁球形,肉质硬而有弹性,稻草或奶油黄色,颜色较均匀;菌褶:稀、不等长,土著白;菌褶:向边缘变薄,白色或淡奶油色;菌柄:长10~35 mm,直径6~11 mm,圆柱形,实心,坚硬,土著白,直生或稍弯曲或纺锤型,基部圆形,味辛辣(见图1.k、图1.l),本实验研究标本:XJ2013100498。

生长环境:生于混交林地上。

该种在亚洲、欧洲和北美洲均有分布,我国主要集中在吉林、海南、云南、甘肃、江苏、湖南、广东、广西、等省。

表1 基于ITSrDNA序列构建红菇属真菌系统树的基因序列信息Tab.1 ITS rDNA specimens of Russula in phylogenetic tree

2.2 分子系统学

基于本研究采集的野生红菇ITSrDNA基因序列排序由508个碱基组成,其中214个碱基有简约信息位点,183个碱基恒定不变,91个碱基无简约信息位点。简约法分析生成5000棵树,Kishino-Hasegawa 测试显示各个聚类树之间没有显著差异,其中一棵如图2所示(最佳聚类树),其树长(Tree length)=878;一致性指数(CI)=0.551;保留指数(RI)=0.714;复定指数(RC)= 0.393;非同源性指数(HI)=0.449。

分子系统学分析由34个种组成,其中包括美红菇Russulapuellaris、鳞盖红菇R.lepida、菱红菇R.vesca、以及绿菇R.virescens、R.melzeri等种类。由图2中可知,绝大多数种都能形成单独的一簇,而且自展支持率较高,只有少数争议种类还待进一步核实形态后做进一步的说明。根据生物条形码原则,ITS rDNA可以作为红菇鉴定的较好的条形码基因。

图2 7种红菇及来自Genbank的最相似物种的ITS序列构建的系统发育树Fig.2 phylogenetic tree of ITS for 7Russula and the similar speices form Genbank

该系统发育树是以GloeocystidiellumaculeatumSheng H. Wu(菌株:AY061739)为外群用PAUP*软件构建的MP系统树。Bootstrap值为1000次重复后高于50%以上的值。由图2可见MYH2013100345号标本与Russulanauseosa,XJ2013100413号标本与R.puellaris,XJ2013100412号标本5与R.lepida,XJ2013100418号标本与R.vesca,XJ2013100520号标本与R.virescens,XJ2013100408号标本与R..faustiana,以及XJ2013100498号标本与R.farinipes均与各自从GenBank检索比对得到的最相似物种以100%的bootstrap值聚在了一起,提示这7种红菇菌株应该各自与其最相似物种为同种的红菇。这一结果基于ITS序列的分子水平上支持了本研究的形态鉴定结果。

3 结论与讨论

本研究表明,综合形态与分子鉴定研究结果,可以确定采自麻阳河自然保护区的7个标本:MYH2013100345号标本鉴定为Russula.nauseosa(厌味红菇),XJ2013100413号标本鉴定为R.puellaris(美红菇),XJ20131004125号标本鉴定为R.lepida(鳞盖红菇),XJ2013100418号标本鉴定为R.vesca(菱红菇),XJ2013100520号标本鉴定为R.virescens(变绿红菇),XJ2013100408号标本鉴定为R.faustiana,以及XJ2013100498号标本鉴定为R.farinipes(粉柄红菇)。

红菇在麻阳河自然保护区分布广泛,具有极高的多样性,是麻阳河野生大型真菌的优势属。但由于其子实体形态特征的分类鉴定不仅受生长环境、生长时期等客观因素的影响,而且形态性状的选取和判断都受主观因素的影响也较大,因而基于形态性状特征对大型真菌进行分类鉴定总是有一定的局限性,特别是对于一些形态特征极为相近的种,仅凭形态极难准确区分鉴定到种一级。而真菌的ITS rDNA区段的保守性基本上表现为种内相对一致、种间差异比较明显,这一特点使得ITS区段不仅适用于属内物种间或种内差异较明显的菌群间的系统发育关系分析,而且非常适合于真菌物种的分子鉴定[20]。郭亮[21]等利用形态学和ITS测序相结合的方法鉴定了湖南松乳菇。Zahhoor[22]等利用形态学和测序方法鉴定了Russulaformula和Russulapostina,。所以,本研究在对麻阳河自然保护区的红菇及其它真菌资源调查的基础上,充分利用ITS快捷、方便、科学的特点[23],借助GenBank等生物数据库的大量信息,从分子水平上为这些红菇真菌资源的准确分类、真菌类群的系统发育提供了科学依据。

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[20] 付立忠,李海波,魏海龙,等. 四个红菇科菌株的rDNA ITS 序列分析和系统发育研究[J]. 食用菌学报,2007,14(2):23~28.

[21] 郭 亮,刘君昂,周国英,等. 基于形态特征和ITS 序列分析对湖南松菌的分类鉴定[J]. 食用菌学报,2011,18(2):15-18.

[22] Zahoor A I, Zafar A R, Khurshid IA,etal. Characterization and identification of Russula formula and Russula postina from Hialayan moist temperate forests of Kashmir[J].AfricanJournalofBiotechnology, 2013,12(23):3643-3647.

[23] 李海波,吴学谦,魏海龙,等. 基于形态特征和ITS序列对7个鹅膏属菌株的分类鉴定[J]. 菌物研究,2007,5(1):14-19.

山地农业生物学报2016年 第35卷 总目次(1~6期)

第1期

·专家特稿·

中国隐脉叶蝉族一新属新种记述(半翅目:叶蝉科:横脊叶蝉亚科)

李子忠,邢济春(1)

·研究报告·

不同药剂处理对猕猴桃根结线虫的防治效果

田雪莲,尹显慧,龙友华,蔡 滔,吴小毛,朱流红(5)

亮甲酚蓝染色法评价斑马鱼卵母细胞体外成熟度的实验研究

方 晨,张 钊,韦祥相,李贤梅,张小雪(9)

白背飞虱交配和产卵行为研究

杨 航,杨 洪,金道超,戴仁怀,周 操(14)

烟草黑胫病菌对7种杀菌剂的敏感性

朱流红,安启菲,吴轶凡,龙友华,赵赞伟,尹显慧(18)

红锥幼树配方施肥效应分析

李宝福,朱 炜,李 莹(23)

大豆中硝磺草酮的残留量测定

王晓环,康 頔,王德飘,吴玉娥,曾 松,张侃侃,胡德禹(29)

食品包装材料中邻苯二甲酸酯向食品模拟物中的迁移规律

曾云想,梁婷婷,汤明河,刘小平(34)

MS-222对杂交鲟幼鱼的麻醉效果及模拟运输试验

王昊龙,徐 弘,安 苗,田茂位,罗慧中(40)

水杨酸对打顶后烟草根系钾吸收的调控作用研究

杨 亚,赵杰宏,姜 骁,母应春,苏 伟(45)

茶树根际促生菌的筛选与促生特性的研究

韩丽珍,邓兆辉,朱春艳,梁彩娇,韦德萍(51)

烟草多酚氧化酶基因的克隆与序列分析

蔡传斌,吴玉俊,曹 宁,龚 露,吴拥军(57)

马铃薯早疫病拮抗菌株筛选及其抗菌活性

杨登路,雷帮星,康冀川,韩 龙(62)

艾纳香属药用植物的化学分类学

丁 健,庞玉新,袁 蕾,张影波,孙懂华,赵 致(67)

·山区开发·

合江方竹的生长研究

马 师,孙巧玲,苟光前(71)

·研究简报·

贵烟2号留叶数对产量和质量的影响

陈毓贤,刁朝强,周建云,胡 勇,韦晓松,宋俊衡,艾复清(74)

不同饲料蛋白来源对犊牛能量及蛋白代谢的影响

刘 燕,杨正德,罗爱平,缪万荣(78)

适于悬浮培养的半夏疏松愈伤组织诱导及增殖技术

刘贵贤, 张明生, 李小兰, 胡珊珊,韦红边,高晓峰(82)

镉胁迫对一年生黑麦草生理生化特性的影响

张凯凯,孙圆圆,徐玲玲,冯旭东,关 萍(86)

风力抑制“遵义红”茶“发酵”研究

申 东,何 萍,申 立,苟廷维,刘思伟,王 勇,王 彤,郑文佳(90)

第2期

·专家特稿·

炭基肥在贵州山地特色高效农业发展中的应用及其效益分析

刘 方,陈祖拥,刘元生,何腾兵(1)

·研究报告·

金铁锁愈伤组织花色苷的稳定性研究

韦红边,李小兰,张明生,刘贵贤,高晓峰,徐佳瑜(7)

半夏及其易混品的DNA分子鉴定

冯 图,何 洋,阮培均(15)

施用氮磷钾肥对白术种子产量和质量的影响

彭方丽,赵 致,刘红昌,王华磊,黄明进,姚伦国(21)

不同烟草品种绿原酸含量变化研究

万 诚,刘仁祥,聂 琼,龙 丽,徐如宏(25)

31%柠檬醛羟丙基-β-环糊精水剂制备及其对稻瘟病菌的抑制作用

高 强,李 明,李荣玉,郑 丽,胡贤峰(29)

杜鹃兰假鳞茎发育中内源激素的含量变化

高晓峰,吕 享,吴彦秋,王汪中,黄图成,韦红边,刘贵贤,张明生(34)

务川耕地土壤养分状况的调查与分析

杨俊松,王德炉,吴春玉,黄付军(40)

三叶木通果实转录组测序初步分析

杨 航,刘红昌,石小兵,葛 菲,赵 致,罗 辉(46)

·山区开发·

花溪国家城市湿地公园景观植物应用

王 建,王秀荣,孙 苏,谭 丽,刘红玉(52)

不同配比氮磷钾硼肥对万寿竹种子发芽的影响

朱 力,王华磊,赵 致,刘红昌,杨振华,曾垂剑,赖雅丽(58)

·文献综述·

AM真菌离体双重培养研究进展

冉海燕,江 龙(62)

·研究简报·

猕猴桃病原真菌拮抗菌筛选

何应红,钱一鑫,康冀川,雷帮星(68)

多效唑和赤霉素对结球白菜抽薹开花的调控

胡存彪,陈云飞,刘圣会,彭剑涛(73)

恩施碎米荠营养成分分析研究

曹 斌,邓国栋(76)

凯里白酸汤的优化工艺

郑莎莎,宋煜婷,罗 意,徐 俐(80)

隆阳烟区植烟土壤养分评价及分类

程昌新,张发明,段迎建,张玉丰,王 超,李 伟,朱海滨,何晓健,张 静,闫 辉,刘 浩(86)

二氢杨梅素对鲤鱼生长及某些生理指标的影响

秦 湉,李芳香,逯凤肖,郁建平(91)

第3期

·专家特稿·

西南玉米生态区域划分及品种需求

陈泽辉(1)

·研究报告·

利用RNA干扰技术沉默稻纵卷叶螟几丁质合成酶A基因

李 娇,杜 娟,李尚伟(10)

槲皮素糖苷类化合物的合成及其对α-葡萄糖苷酶的抑制活性

文中行,胡占兴,袁 洁,陈洪菊,张利梅,梁光义,徐必学(18)

云南扁萤幼虫捕食功能研究

吴健卓,郭 琼,杨毓银,陈文龙(25)

转拟南芥BAS1基因提高烟草光合特性

姚新转,吕立堂,赵德刚(30)

开放式和封闭式TBS捕鼠效果观察

韦应敏,留 青,杨德辉,李定超(35)

云烟87大小孢子形成和雌雄配子体发育观察

吉志超,李祖任,高凯悦,张依楠,廖海民(39)

3-芳基-2-硝基丙醇的合成新方法

梁 弘,李齐激,杨 艳,潘 雄,杨 娟,杨小生(44)

黄精产地加工工艺研究

杨顺龙,周 英,赵 致,林 冰(49)

·山区开发·

基于生态足迹模型的贵州省

森林公园旅游可持续发展研究

李俊峰,聂朝俊(53)

贵州省江口县木本药用植物种质资源调查

王 瑶,苟光前,孙巧玲,陈云飞(58)

·研究简报·

小黄姜组培快繁技术研究

黄 明,赵懿琛,赵德刚(63)

黑果枸杞光合作用日变化规律研究

马彦军,马 瑞,马玉祥,李情辉(66)

毛红椿木材物理力学性质研究

叶忠华,叶友章,刘晓辉,杨淑宝(72)

山核桃蒲壳含量对金森女贞容器育苗的影响

金久宏(76)

两种低聚糖对烟草种子萌发及幼苗生长的影响

余 钱,尹显慧,陈 雪,黄化刚,代园凤,喻会平(79)

二氢杨梅素体外抗氧化活性研究

王恩花,杨礼寿,逯凤肖,秦 湉,秦泽华,郁建平(86)

一株产红色素的丝枝蜡蚧霉新菌株

董 旋,曾桂萍,陈万浩,韩燕峰,梁宗琦(91)

第4期

·专家特稿·

贵州烤烟育种工作进展与对策探讨

刘仁祥,夏志林,崔庆伟,喻奇伟,张庆明,聂 琼,徐秀红,刘 洋(1)

·研究报告·

中国巨螯螨科一新种和三新记录种记述暨名录增订(蜱螨亚纲:中气门目)

黄振兴,金道超,乙天慈(6)

贵州紫苏叶中氨基酸及游离氨基酸含量分析

金石诚,刘忠伟,沈 奇,李 月,牛 熙,朱秋劲(13)

黑骨藤追风活络微丸处方及制备工艺的研究

乔 杨,王慧娟,彭礼军,谭 婕,李 俐(18)

基于转录组信息的艾纳香牻牛儿基牻牛儿基焦磷酸合成酶基因(BbGGPS) 的克隆及序列分析

夏奇峰,赵 致,刘红昌,官玲亮,庞玉新(23)

岩溶地区山地城市山体绿地资源公园化利用初探

张 远,王志泰(30)

黄壤小流域水田和旱地土壤养分差异性探析

易 香,焦克强,段建军,卜通达,熊永春,周 艳,陈祖拥(36)

三峡库区小流域紫色土玉米推荐施肥指标体系研究

唐晓东,陈燕霞,张树全,王正银,屈 明(42)

贵州省江口县常用木本纤维植物资源初步调查

陈云飞,苟光前,王 瑶,孙巧玲,王 灯(48)

·山区开发·

基于风险预测的山地典型农业生长期干旱指标筛选

侯双双 , 谷晓平(54)

不同覆盖方式对茶园土壤水分及茶叶产量的影响研究

胡承兴,舒英格,何 秀(61)

·文献综述·

酒糟的保存和应用现状

李芳香,郁建平(66)

·研究简报·

抗癌肽改造及其对人乳腺癌MCF7作用的评估

党军龙,周礼红(72)

不同叶面肥对狐臭柴叶片生物量和主要生化指标的影响

颜跃跃,王汪中,吕 享,李小兰,黄图成,张明生(77)

Box-Behnken 设计优化贵州野生地枇杷总黄酮提取工艺

陈 琳,俸婷婷,游 婵,李 超(83)

光谱判断田间鲜烟叶成熟度的影响因子研究

徐兴阳,何春荣,秦春丽,王 锋,劳彩莲(88)

第5期

·专家特稿·

贵州省发展优质米产业浅见

余显权(1)

·研究报告·

杜仲EuCHIT1基因表达载体构建及在大肠杆菌中的表达

董 旋,丁延庆,冉 昕,赵德刚(9)

野百合根系水浸液对几种花卉的化感效应

胡小京,代玉苹(13)

宽阔水自然保护区蕨类植物初步研究

孙巧玲,苟光前,王 瑶,陈云飞,王 灯(18)

贵州黑山羊IL-2基因的克隆及序列分析

胡贵川,冯 将,鲜思美,刘 嫒,李鹏飞,肖拉拉(25)

水稻缙恢21诱变材料遗传变异的SSR分析

张修宝,罗洪发,查仁明,冯仕飞,樊海容,张姗姗,庹中丽(30)

贵州省大方县老鹰岩地区苔藓植物研究

赵 虹,熊源新(37)

·山区开发·

基于苗族文化特色的山体公园景观设计

——以安顺市跳花坡山体公园景观设计为例

冯 丽,李美君,白新祥,张 然,谢 伟,周虹宏(43)

山地梯田湿地景观设计探讨

——以瓮安水瓮田梯湿地公园景观设计为例

冯凤娇,李季璇,唐轶凡,任芷妮,赵 丹,何嵩涛(51)

·文献综述·

水稻主要粒型基因及其遗传调控的研究进展

涂 坦,付 洪(58)

·研究简报·

黔产紫苏挥发油提取方法优化及化学成分分析

张 敏,周 英,骆 静,陈 琳,俸婷婷(66)

贵州黔陶乡古茶树茶品质研究

汪艳霞,李 伟,钟明跃,尹 杰(70)

贵丰优785产量与主要农艺性状的相关分析

蔡 源,黄宗洪,甘 雨,向关伦,杨占烈,郭 慧,潘建慧,龙江辉(73)

粉花绣线菊引种前后ATISINE型二萜生物碱含量比较研究

何俊梅,苑春茂,陈永霞,郝小江,张朝阳(77)

黑豆汁的制备工艺及质量控制指标的研究

黄 倩,廖 力,陈胜利,廖光星, 林 冰(83)

二氢杨梅素对菜籽油和花生油氧化作用的影响研究

王恩花,杨礼寿,逯凤肖,郁建平(89)

第6期

·专家特稿·

不同类型猴樟混交林种间关系和冠层结构

韦小丽,陈瀚林(1)

·研究报告·

中国始叶螨属区系特征分析

闫德川,乙天慈,郭建军,金道超(9)

秦岭烟区不同基因型烤烟化学成分与挥发性香气物质研究

樊文举,高娟娟,王玉堂,刘拉平,杜双奎,张立新,金保锋,扈 强,张建新(18)

EDTA对镉胁迫下紫云英种子萌发及幼苗生理抗性的影响

杨 鹏,陈兴银,田玲玲,张凯凯,徐玲玲,关 萍(26)

8种药剂对烟草黑胫病菌室内毒力测定

高亚星,彭丽娟,蒋选利(31)

濒危植物珙桐大小孢子形成和雌雄配子体发育观察

陈兴银,张玉晶,张凯凯,杨 鹏,石建明(35)

·山区开发·

贵州省江口县木本蜜源植物资源初步调查

陈云飞,苟光前,王 瑶,孙巧玲,王 灯(40)

不同基质对冬青叶鼠刺容器苗生长的影响

敖国富,周 玮,常 秋,梁 娥,熊仕俊,吕 涛,周双宝(49)

·文献综述·

UV辐射对植物多酚代谢的影响

李 雪,潘学军,张文娥(54)

野生稻种质在水稻抗病育种上的研究及应用进展

王樟凤,盛文涛,柴学文,饶友生(61)

·研究简报·

三种茶香猪肉干品质的比较分析

钟明跃,魏正松,李晓松,尹 杰,汪艳霞(68)

凝结芽孢杆菌提高放牧黔北麻羊断奶羔羊生长和养分消化的研究

陈 坚,张 蓉,施晓丽,龙 杰,罗卫星(73)

不同土壤消毒剂对药用百合病害防治效果的研究

周佳民,宋 荣,张天术,朱校奇,曹 亮,彭斯文(76)

首乌藤不同部位有效成分含量考察

廖 力,黄 倩,陈胜利,廖光星,林 冰(81)

贵州麻阳河自然保护区7种野生红菇的形态与分子鉴定

谯利军,文庭池,康冀川,康 超,吴兴亮(87)

Journal of Mountain Agriculture and Biology
General Contents of the Vol.35 2016(No.1-6)

No.1

·Featured Articles·

A new genus and a new species of the tribe Nirvanini (Hemiptera: Cicadellidae: Nirvaninae) from China

LI Zi-zhong, XING Ji-chun(1)

·Research Papers·

Control effect of different pesticide treatments against root-knot nematodes onActinidia

TIAN Xue-lian, YIN Xian-hui, LONG You-hua, CAI Tao, WU Xiao-mao, ZHU Liu-hong(5)

An experimental study on the evaluation of thematurity of zebrafish oocytesin vitro using brilliant cresyl blue staining

FANG Chen, ZHANG Zhao, WEI Xiang-xiang, LI Xian-mei, ZHANG Xiao-xue(9)

Study on the mating and oviposition behavior ofSogatellafurcifera(Horváth)[Hemiptera:Delphacidae]

YANG Hang, YANG Hong, JIN Dao-chao,DAI Ren-huai, ZHOU Cao(14)

Determination on Sensitivity ofPhytophthoraNicotianaetoSeven Fungicides

ZHU Liu-hong, AN Qi-fei, WU Yi-fan, LONG You-hua, ZHAO Zan-wei, YIN Xian-hui(18)

Effectof formulated fertilizer on the saplings ofCastanopsishystrix[Fagales: Fagaceae]

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Identification and Characterization of Plant Growth-promoting Rhizobacteria Isolated from Rhizosphere soils of Tea Trees

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The Screening of Antagonistic Strains to Potato Early Blight and Detection of Antimicrobial Activity

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CHEN Yu-xian,DIAO Chao-qiang,ZHOU Jian-yun,HU Yong,WEI Xiao-song,SONG Jun-heng,AI Fu-qing(74)

Effects of Different Protein Sources for Energy and Protein Metabolism in Calves

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Effects of cadmium stress on physiological and biochemical characteristics inTagetespatulaL. [Asterales: Asteraceae]

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WThe Inhibitory Effect of Wind on the Fermentation of Zunyi Black Tea

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No.2

·Featured Articles·

The Application of Biochar-fertilizer and Its Benefit Analysis for Developing Efficient Agriculture with Mountainous Characteristic in Guizhou Province

LIU Fang,CHEN Zu-yong, LIU Yuan-sheng, HE Teng-bing(1)

·Research Papers·

Studies on the Stability ofAnthocyanin inPsammosilenetunicoidesCallus

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Identification ofPinelliaternataand Adulterants Based on DNA Barcoding

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Influence of Fertilization on the Yield and Quality ofthe Seed ofAtractylodesmacrocephala

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Dynamic Changes of Chlorogenic Acid Content in Different Types of Tobacco Cultivars

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Preparation of Citral HP-β-CD 31% AS and Its Biological Activity of Magnaporthe Grisea

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Endogenous Hormone Content Changes During NewbornCremastraPseudobulb Development

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Investigation and Analysis on Soil Nutrient Status of Main Cultivated Land in Wuchuan County

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Transcriptome Analysis ofAkebiatrifoliatein Ripening Process

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·Mountain Area Development·

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·Literature Review·

The research progress of AM fungi in vitro dual culture

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·Short Communications·

Screening of Antagonistic Fungi against Pathogenic Fungi of Kiwifruit and Determination of its Antimicrobial Activity

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Regulation of Flowering Time of Chinese Cabbage by Paclobutrazol and Gibberellin

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Analysis of Nutritive Ingredients inCardamineenshiensis

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Optimization ofthe Technology of Kaili White Sour Soup by Orthogonal Test

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Classification and Evaluation over SoilNutrient Content in Longyang Tobacco-growing Areas

……CHENG Chang-xin, ZHANG Fa-ming, DUAN Ying-jian, ZHANG Yu-feng, WANG Chao, LI Wei, ZHU Hai-bin, HE Xiao-jian, ZHANG Jing, YAN Hui, LIU Hao(86)

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No.3

·Featured Articles·

Ecological regions and variety requirements of the southwest corn

CHEN Ze-hui(1)

·Research Papers·

Using RNA interference technique to silence chitin synthase A Gene in the rice leaf folder, Cnaphalocrocis medinalis (Lepidoptera: Pyralidae)

LI Jiao, DU Juan, LI Shang-wei(10)

Synthesis of Quercetin Glycosides and Their Inhibitory Activities to α-Glucosidase

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Study on the predatory function of larva ofLampyrigerayunnana(Fairmaire) [Coleoptera: Cantharidae]

WU Jian-zhuo, GUO Qiong, YANG Yu-yin, CHEN Wen-long(25)

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JI Zhi-chao, LI Zu-ren, GAO Kai-yue, ZHANG Yi-nan, LIAO Hai-min(39)

A new Method for Synthesis of 3-aryl-2-nitropropanol

LIANG Hong, LI Qi-ji, YANG Yan, PAN Xiong, YANG Juan,YANG Xiao-sheng(44)

Optimization of Processing Technologiesin Origin Place ofPolygonatumCyrtonema

YANG Shun-long,ZHOU Ying,ZHAO Zhi,LIN Bing(49)

·Mountain Area Development·

Study on the sustainable development of tourism in forest parks of Guizhou Province based on the ecological footprint model

LI Jun-feng,NIE Chao-jun(53)

Investigation of woody medicinal plant resources in Jiangkou County of Guizhou Province

WANG Yao, GOU Guang-qian,SUN Qiao-ling,CHEN Yun-fei(58)

·Short Communications·

Rapid Propagation ofLiu Panshui Small Yellow Ginger

Ming Huang ,Yi-Chen Zhao,De-Gang Zhao(63)

Studies on diurnal change of photosynthesis of Lycium ruthenicum [Solanales: Solanaceae]

MA Yan-jun, MA Rui,Ma Yu-xiang, LI Qing-hui(66)

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YE Zhong-hua,YE You-zhang,LIU Xiao-hui,YANG Shu-bao(72)

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JIN Jiu-hong(76)

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YU Qian,YIN Xian-hui,CHEN Xue,HUANG Hua-gang,DAI Yuan-feng,YU Hui-ping(79)

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No.4

·Featured Articles·

Progress in Tobacco Breedingin Guizhou Province and Discussion for Solutions

LIU Ren-xiang, XIA Zhi-lin, CUI Qing-wei, YU Qi-wei, ZHANG Qing-ming, NIE Qiong, XU Xiu-hong, LIU Yang(1)

·Research Papers·

Description of a new species of Macrochelidae (Acari: Mesostigmata) and update of the species checklist of the family with three new records from China

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JIN Shi-cheng, LUI Zhong-wei, SHEN Qi, LI Yue, NIU Xi, ZHU Qiu-jing(13)

Study on the formulation and preparation of a mixed Chinese herbal medicine - Heiguteng Zhuifeng Huoluo Pellets

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Analysis of geranylgeranyl pyrophosphate synthase (GGPS) sequence ofBlumeabalsamifera(L.)·DC based on transcriptome information

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·Mountain Area Development·

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·Literature Review·

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LI Fang-xiang ,YU Jian-ping(66)

·Short Communications·

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YAN Yue-yue, WANG Wang-zhong, LV Xiang, LI Xiao-lan, HUANG Tu-cheng, ZHANG Ming-sheng(77)

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XU Xing-yang,HE Chun-rong,QIN Chun-li,WANG Feng, LAO Cai-lian(88)

No.5

·Featured Articles·

Suggestions on Production of High Quality Rice in Guizhou Province

YU Xian-quan(1)

·Research Papers·

Construction ofEucommiaulmoidesEuCHIT1 gene expression vectors and its expression inEscherichiacoli

DONG Xuan,DING Yan-qing,RAN Xin,ZHAO De-gang(9)

The Allelopathic Effects ofLiliumbrowniiRoot Exudates on Some Ornamental Plants

HU Xiao-jing,DAI Yu-ping(13)

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SUN Qiao-ling, GOU Guang-qian, WANG Yao,CHEN Yun-fei,WANG Deng(18)

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HU Gui-chuan,FENG Jiang,XIAN Si-mei,LIU Ai,XIAO La-la(25)

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·Mountain Area Development·

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·Literature Review·

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TU Tan, FU Hong(58)

·Short Communications·

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Effect of Dihydromyricetin on oxidation of rapeseed oil and peanut oil

WANG En-hua,YANG Li-shou,LU Feng-xiao,YU Jian-ping(89)

No.6

·Featured Articles·

Theintraspecific relationship and canopy structure of different typeCinnamomumbodinierimixed stand

WEI Xiaoli, CHEN Hanlin(1)

·Research Papers·

The Faunal Analysis ofEotetranychusfrom China

YAN De-chuan, YI Tian-ci, GUO Jian-jun, JIN Dao-chao(9)

Study on Chemical Components and theVolatile Aroma in Different Genotypes of Flue-cured Tobacco in Qinling Area

FAN Wen-ju, GAO Juan-juan, WANG Yu-tang, LIU La-ping, DU Shuang-kui, ZHANG Li-xin,JIN Bao-feng, HU Qiang, ZHANG Jian-xin(18)

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Toxicological Test of Eight Fungicides to Tobacco Black Shank

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·Mountain Area Development·

Preliminary Investigation of Woody nectar plant resources in Jiangkou County

Chen Yun-fei,Gou Guang-qian,Wang Yao, Sun Qiao-ling,Wang Deng(40)

Effects of Different Substrates on Growth of container seedlings ofIteaIlicifolia

AO Guo-fu, ZHOU Wei, CHANG Qiu, LIANG E, XIONG Shi-jun, LV Tao, ZHOU Shuang-bao(49)

·Literature Review·

Effect of UV Radiation on the Metabolism and the Pathway of Plant Polyphenol

LI Xue, PAN Xue-jun, ZHANG Wen-e(54)

Advances on Disease-resistant Breeding Utilized with Wild Rice Germplasm Resources in Rice

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·Short Communications·

Comparative analysis of three kinds of tea quality of dried pork

ZHONG Ming-yue, WEI Zheng-song, LI Xiao-song, YIN Jie, WANG Yan-xia(68)

The Improvements of Growth Performance and Nutrients Digestibilitiy on Grazing Qian-bei-ma Weaned Lambs fedBacillusCoagulans

CNEN Jian, ZHANG Rong,SHI Xiao-li,LONG Jie,LUO Wei-xing(73)

Control effects of soil disinfectants onLiliumlancifoliumThunb disease

ZHOU Jia-rnin, SOG-Rong,ZHANG Tian-shu, ZHU Xiao-qi, CAO-Liang,PENG Si-wen(76)

Study of effective components in different parts ofCaulisPolygoniMultiflori

LIAO Li, HUANG Qian , CHEN Sheng-li , LIAO Guang-xing, LIN Bing(81)

Morphology and phylogeny of sevenRussulaspecies from Mayang River National Nature Reserve of Guizhou

QIAO Li-jun, WEN Ting-chi, KANG Ji-chuan, KANG Chao,WU Xing-ling(87)

Morphology and phylogeny of sevenRussulaspecies from Mayang River National Nature Reserve of Guizhou

QIAOLi-jun1,WENTing-chi1,KANGJi-chuan1*,KANGChao2,WUXing-ling2,3

(1.EngineeringResearchCenterofSouthwestBio-PharmaceuticalResources,MinistryofEducation,GuizhouUniversity,Guiyang,Guizhou550025,China;2.GuizhouAcademyofSciences,Guiyang,Guizhou570228,China)

This study provided a clue for protection and utilization ofRussulaspp. inMayang River reserve and Guizhou Province. Seven species ofRussulaspp. were collected from the Mayang River Nature Reserve in Guizhou Province, China. These species were identified as theR.nauseosa、R.puellaris、R.lepida、R.vesca、R.virescensandR.faustianabased on morphology and phylogenetic analysis of ITS rDNA sequences.

Russulaspp; Morphological Classification; Molecular Identification; ITS rDNA

2016-08-31;

2016-09-23

科技基础性工作专项项目(2014FY120100);贵州省科技厅创新能力建设项目(黔科合G(2010)4009);贵州大学青年科技基金项目(No. 201309)。

Q945.78

A

1008-0457(2016)06-0087-06 国际

10.15958/j.cnki.sdnyswxb.2016.06.015

*通讯作者:康冀川(1962-),男,博士,教授,主要研究方向:药用真菌资源;E-mail:jckang@gzu.edu.cn。

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