春兰菌根菌的分离及共生培养体系的研究

采用根组织切片法自野生春兰菌根共分离到6株真菌菌株,接种春兰杂交组培苗,结果发现CL-3和CL-6菌株在菌-苗共生培养基Ⅰ中对幼苗生长有不同程度的促进作用,2 个月后处理苗的鲜重增长率分别达到47.4%和42.5%,与对照差异达显著水平(P＜0.05).在CL-3和CL-6接菌处理苗的营养根中均分离获得了原接种真菌,并观察到了菌根结构.结果表明,CL-3和CL-6菌株已与组培幼苗成功建立了共生培养体系.     

2种杓兰属植物内生狭截盘多毛孢菌的分离与鉴定

该文报道了西藏杓兰(Cypripedium tibeticum)和无苞杓兰(C. bardolphianum)根中内生真菌新记录种——狭截盘多毛孢(Truncatella angustata)。该研究从四川黄龙沟不同海拔区分别采集到西藏杓兰和无苞杓兰,并在其根中分离获得148个狭截盘多毛孢菌株。形态学观察显示,狭截盘多毛孢菌株在PDA培养基上菌落白色,背面黄褐色;分生孢子器黑色脓包状;分生孢子梭型,4个细胞,3个隔膜;顶端附属丝1~5根。ITS rDNA分析发现3种新基因型菌株(HLIO15_15a_22、HLIO15_20a_42和HLIO15_17a_46);在海拔最高(3 330~3 400 m)的居群中狭截盘多毛孢的分离率最高,且包含全部3种基因型菌株。构建截盘多毛孢属(Truncatella)真菌ITS序列系统发育树,发现狭截盘多毛孢不同生态功能的菌株没有形成明显分支。     

云南金铁锁根腐病病原菌的分离及鉴定

金铁锁(Psammosilene tunicoides)是西南地区重要的民族民间药物、云南白药等中成药的主要原料药。为分离并鉴定云南金铁锁根腐病病原菌,该研究采用微生物纯培养方法,分离纯化金铁锁根腐病植株根部的病健交接处组织,获得金铁锁根腐病病原微生物。按照Koch''s法则验证分离得到的病原菌的致病性。结合形态学观察、真菌rDNA-ITS和TEF-1α序列分析以及系统发育树的构建鉴定获得的病原菌的种类。结果表明:(1)从金铁锁根腐病植株组织中分离纯化得到85株真菌,其中PSD-1、PSD-2、PSD-3菌株均能引起金铁锁的根腐病。(2)金铁锁健康植株接种PSD-1、PSD-2、PSD-3菌株后,产生与大田植株根腐病相似的症状,并且发病率分别为60%、61.7%、71.7%。(3)结合形态学观察及基因序列分析,鉴定该三株菌株均为尖孢镰刀菌(Fusarium oxysporum)。该文首次报道了尖孢镰刀菌是引起金铁锁根腐病的主要病原菌之一,为深入探究具有根腐病生物防治功能的内生菌群及其作用机制奠定了基础,也为后续金铁锁病害的防治工作提供了理论依据。     

云南嵩明大哨天然虫生真菌及其内生真菌的分离鉴定与抑菌活性分析

【背景】云南存在着丰富的虫草资源,天然虫草群落中蕴藏多样的真菌资源,是挖掘新型抑菌活性化合物的潜在来源。【目的】了解云南嵩明大哨天然虫生真菌及其内生真菌的物种多样性,并从中筛选具有抑菌活性的菌株。【方法】采用组织分离法对所采集的野生虫草样本进行虫生真菌及其内生真菌的分离,并通过形态观察和ITS联合nrSSU、nrLSU、tef-1α、rpb1、rpb2多基因测序进行物种鉴定及多样性分析;通过平板对峙法以7株病原细菌、5株植物病原真菌为病原指示菌进行抑菌活性测试。【结果】共采集86份天然虫草样本,经鉴定隶属于3科5属7种,包括虫草科（Cordycipitaceae）虫草属（Cordyceps）（1种）、白僵菌属（Beauveria）（2种）、鳞翅虫草属（Samsoniella）（2种）、麦角菌科（Clavicipitaceae）泛普可尼亚属（Metapochonia）（1种）,以及线虫草科（Ophiocordycipitaceae）线虫草属（Ophiocordyceps）（1种）。同时,从采集到的虫草样本中分离得到26株内生真菌,分属于9科9属,其中木霉属（Trichoderma）（38%）和镰刀菌属（Fusarium）（19%）为此次分离得到的优势菌属。对所分离保存的真菌按其分离源及种类归属,从虫生真菌和内生真菌菌株中共挑取20株代表性菌株进行抑菌活性筛选,有11株真菌对2株及以上病原指示细菌具有不同程度的抑菌活性,13株真菌对1株以上植物病原真菌具有不同程度拮抗能力;其中Trichoderma sp.Y3-1和Fusarium sp.WZ3-1具有广谱抑菌活性。【结论】云南嵩明大哨分布有丰富的虫生真菌及内生真菌资源,分离所得的真菌对多种病原菌具有不同程度的抑制作用。本研究丰富了云南虫草资源多样性,为云南省虫草及其内生真菌资源的开发利用提供了数据支持,也为下一步从虫草及其相关真菌资源中挖掘抑菌活性物质提供了菌株资源。     

山西省不同地区柴胡根部可培养内生真菌的多样性分析

为研究柴胡（Bupleurum chinense DC.）根部可培养内生真菌的多样性，分析山西省不同地区柴胡根部可培养内生真菌多样性的差异性，建立柴胡根部内生真菌库。本文以山西省不同产地的柴胡根为材料，采用传统植物组织平板分离法对柴胡根部可培养内生真菌进行分离培养和分子鉴定。从25个不同产地的柴胡样品根部中共分离培养得到705株真菌，经分子生物学鉴定，其中695株真菌归属为4门14纲23目32科55属119种。晋南地区的优势菌属为镰刀菌属（Fusarium, 32.18%）、亚隔孢壳属（Didymella, 15.96%）、曲霉属（Aspergillus, 8.24%）、异茎点霉属（Paraphoma, 6.91%）、茎点球属（Phoma, 5.32%）、链格孢属（Alternaria, 5.05%），晋北地区的优势菌属为镰刀菌属（31.66%）、曲霉属（15.36%）、链格孢属（9.09%）、亚隔孢壳属（8.15%）、异茎点霉属（7.52%），亚隔孢壳属、曲霉属、链格孢属等优势属的相对丰度差异较大；晋南地区的优势物种为Didymella bellidis（11.44%）、三线镰刀菌（Fusarium tricinctum, 7.98%）、腐皮镰刀菌（Fusarium solani, 7.45%）、锐顶镰孢菌（Fusarium acuminatum, 5.85%）、烟曲霉（Aspergillus fumigatus, 5.85%），晋北地区的优势物种为锐顶镰孢菌（14.11%）、烟曲霉（11.91%）、三线镰刀菌（9.40%）、菊异茎点霉（Paraphoma chrysanthemicola, 5.64%）、D. bellidis（5.64%）、链格孢菌（Alternaria alternata, 5.64%），D. bellidis、锐顶镰孢菌、烟曲霉等优势种的相对丰度差异较大。差异性分析表明晋南地区的生物多样性显著高于晋北地区，D. bellidis、锐顶镰孢菌、烟曲霉等优势种的相对丰度差异较大，为进一步研究柴胡根内生真菌相对丰度差异较大的物种及其不同菌株对柴胡皂苷积累的影响提供参考。     

云南三个地区马铃薯内生真菌多样性研究

为研究云南马铃薯(Solanum tuberosum)内生真菌的多样性,该文以采自云南省德宏芒市、大理喜洲和临沧双江3个地区的马铃薯植株为研究对象,采用组织块分离法、尖端菌丝挑取法对马铃薯根、茎及块茎中的内生真菌进行分离纯化,并采用形态学鉴定方法和ITS序列分析法对分离得到的内生真菌进行鉴定,对内生真菌的定殖率、分离率及多样性指数进行计算和分析。结果表明:(1)共分离得到内生真菌98株,其中从德宏芒市的样品中获得40株,从大理喜洲的样品中获得27株,从临沧双江的样品中获得31株。(2)经鉴定,分离得到的马铃薯内生真菌共涵盖10目10科13属,大多为子囊菌门和担子菌门,优势菌为镰刀菌属(Fusarium)和青霉属(Penicillium),褶皱裸孢壳(Emericella rugulosa)、接骨木镰刀菌(Fusarium sambucinum)、毛韧革菌(Stereum hirsutum)、Psathyrella sulcatotuberculosa和Epicoccum catenisporum 5种真菌均为首次从马铃薯植株中分离得到。(3)马铃薯块茎内生真菌的定殖率最高,根部内生真菌定殖率最低; 内生真菌的分离率以马铃薯根部为最高,而茎部最低; 不同组织中内生真菌的多样性指数趋势均为根>块茎>茎。从综合来看,云南马铃薯植株中的内生真菌具有较高的多样性,不同地区的马铃薯样品中内生真菌优势菌不同,马铃薯根部具有最丰富的内生真菌种群和最高的分离率,是最适合进行内生真菌分离的材料。该研究结果为后期探究马铃薯内生真菌对病原菌的拮抗作用奠定了基础,也为马铃薯内生真菌多样性研究提供了参考数据。     

2种地衣共生蓝藻的分离鉴定及其形成藻结皮条件的研究

以2种地衣——地卷(Peltigera rufescens)和平盘软地卷(Peltigera elisabethae)为研究材料,分离培养地衣共生藻,通过形态特征的观察结合rDNA ITS序列分析确定其分类地位,并对地衣共生藻在试验室模拟条件下形成人工藻结皮的优化条件进行分析,为地衣共生藻资源的开发和生产实际利用提供依据。结果表明：(1)从平盘软地卷和地卷分离的藻株I₁b和 L₂均属于蓝藻,基于ITS序列构建的系统树分析结果显示, I₁b与具鞘微鞘藻(Microcoleus vaginatus)的支持率高达100%,属于具鞘微鞘藻;L₂与念珠藻(Nostoc sp.)的支持率为93%,属于念珠藻属。(2)试验室模拟荒漠极端环境中形成人工结皮的优化条件为：沙土含水量10%、接种3∶1混合藻(具鞘微鞘藻:念珠藻)、藻接种量10 μg/cm²。     

葡萄生单轴霉菌围可培养微生物多样性及潜在生防菌研究

【目的】揭示葡萄生单轴霉(Plasmoparaviticola)菌围可培养细菌和真菌的多样性特征,筛选对葡萄霜霉病有较强稳定防治效果的生防菌。【方法】连续两年从我国南北方具有代表性的7个葡萄产区采集葡萄霜霉病叶,镊子夹取经保湿培养获得的新鲜霉层并配制孢子囊悬浮液,采用传统分离培养法,结合形态分类、BOX-PCR指纹图谱分析以及分子鉴定结果,对葡萄生单轴霉菌围的可培养细菌和真菌进行聚类分析;采用菌株及其发酵液与病原菌孢子囊悬浮液等体积混合培养测定其对孢子囊的抑制作用,离体叶片接种法检测该菌株及其发酵液对霜霉病的防治效果。【结果】分离获得了90株细菌和110株真菌,分别归属于8个细菌属和14个真菌属,且相同地区不同葡萄品种葡萄生单轴霉菌围的细菌和真菌在同年处于同一分支。假单胞菌属(Pseudomonas)和枝孢属(Cladosporium)稳定存在于各地区不同品种葡萄霜霉病叶上葡萄生单轴霉菌围;在两年间稳定存在的菌株占比多数在80.0%以上且均具有较高的生防作用;其中,广泛分布的6株枝顶孢属(Acremonium)真菌对葡萄霜霉病的防治效果均较好,最高可达100.0%;防治效果较高的11个菌株的无菌发酵液中,黑曲霉(Aspergillusniger) NX2F、苋楔孢黑粉菌(Thecaphora amaranthi) BJ1G和匍枝根霉(Rhizopus stolonifer) BM1L的无菌发酵液防治效果均为100.0%。【结论】葡萄生单轴霉菌围的可培养细菌和真菌群落主要受地区因素影响,有较高的稳定性和生防作用,揭示了枝顶孢属真菌在我国葡萄主要产区葡萄生单轴霉菌围附生的普遍性,为葡萄霜霉病的防治提供了丰富和宝贵的资源。     

入侵植物紫茎泽兰叶内生真菌多样性研究

外来入侵植物通常要和当地的生物发生相互关系并对当地生态系统产生影响。为表明叶内生真菌是否有利于紫茎泽兰（Eupatorium adenophorum）的入侵力,首次调查了其叶内生真菌多样性。利用过夜冰冻组织法和常规的麦芽汁琼脂平板培养法,分别于3、6月份从昆明西山、金殿种群分离得到312个菌株,根据其ITS基因差异,分为77个可操作分类单元（OTUs）,系统发育地位分布在Agaricomycetes、Dothideomycetes、Sordariomycetes和 Pezizomycetes 4个纲。在低阶分类上这些真菌可分为19个属,优势属为链格孢属（Alternaria）（25.09%）和炭疽菌属（Colletotrichum）（10.80%）。有2.09%的菌株属于未知真菌。相比较,6月份的紫茎泽兰种群比3月份具有较高的内生真菌多样性。结果表明,入侵植物紫茎泽兰叶内生真菌非常丰富,这些真菌是否对紫茎泽兰的入侵力具有影响值得今后深入研究。     

新疆伊犁野生阿魏菇菌株的分离鉴定与培养特性

对采自新疆伊犁的两种具有不同形态特征的野生阿魏菇（Pleurotus ferulae）子实体分离的菌株进行鉴定和系统发育分析,并探讨优良菌株的培养特性。结合子实体形态和生境特征,利用rDNA-ITS序列,对伊犁野生阿魏菇菌株YL-A1和YL-A2进行分子鉴定及系统发育分析。对阿魏菇菌株YL-A1生长温度、pH、碳源、氮源和无机盐进行单因素试验,分析各因素对菌丝生长的影响,通过L₉(3⁴正交试验,筛选各因子之间的最优组合。结果表明,野生阿魏菇菌株YL-A1（ITS序列GenBank登录号：MN460317）和YL-A2（ITS序列 GenBank登录号：MN809132）,不仅子实体形态特征有差异,且菌株YL-A1菌丝生长速度和长势优于YL-A2,并相互拮抗。结合生境和形态特征及rDNA ITS序列分析,将二者鉴定为阿魏菇(Pleurotus eryngii var. ferulae)。系统发育分析发现,菌株YL-A1和YL-A2单独聚为一组,与其他阿魏菇菌株有一定的遗传距离。经单因素和正交试验,确定了菌株YL-A1在含葡萄糖20 g/L、蛋白胨4 g/L、KH₂PO₄ 1 g/L,pH 7.5的培养基中,25 ℃培养,菌丝生长状态最佳,确定了优良菌株YL-A1菌丝生长最佳培养条件。为进一步保护利用伊犁野生阿魏菇菌种资源提供参考。     

春兰内生真菌的分离及其抑菌活性的初步研究

利用内蒙古锡林浩特国家气候观象台1994～2009年牧草生长季逐月实测资料,对CENTURY模型进行检验,模拟内蒙古典型草原1953～2010年间地上净初级生产力（ANPP）动态,并与26个气象因子进行相关性分析。模型检验结果显示,模拟值与观测值之间的相关系数为R²=0.66,斜率b=0.95,误差平方根值为50.51 g·m^-2,平均绝对百分比误差为44.19%。结果表明：（1）CENTURY模型能比较准确地模拟这类草原的季节动态和年际变化;在过去的58年中,内蒙古典型草原温度增加,降水减少,ANPP下降;ANPP变化趋势与降水量相似。（2）用实际气象观测资料模拟获得的ANPP随气温和降水的变化呈现出明显的变化规律,生长季内地上生物量对降水和温度的季节性分布也非常敏感;相关分析进一步表明,ANPP对生长季内降水量和极端高温非常敏感,而与年极端最低气温、平均地面温度、日照时数、平均风速和最大积雪深度无显著相关关系;过去58年研究区ANPP下降是降水减少、温度升高以及干旱事件频发共同作用的结果。（3）根据预测,在SRES B2情景下,未来50～100年内蒙古典型草原生长季平均最高气温和最低气温都将呈升高趋势,2080s分别升高4.01℃、4.35℃,每10年增加速率分别为0.35℃和0.38℃;降水量略呈增加,2020s、2050s和2080s研究区生长季将分别增加3.17%、5.13%和7.03%,每10年增加速率为0.09 mm;ANPP呈下降趋势年际间波动较大,2020s、2050s和2080s研究区将分别下降5.76%、7.52%和11.42%,每10年下降速率为0.76 g·m^-2。     

Genetic diversity of wild Cymbidium goeringii (Orchidaceae) populations from Hubei based on Inter-simple sequence repeats analysis

Cymbidium goeringii is a diploid and nonrewarding, bumblebee-pollinated species, which is distributed in China, Japan and Korea Peninsula. This species is now highly endangered due to the mass collection and forest clearance in China. In the present study, we investigated the distribution of genetic variation within and between eleven populations of Cymbidium goeringii in central China by using Inter-simple sequence repeats (ISSR) markers. Eleven primers produced a total of 127 clear and reproducible bands of which 112 were polymorphic. High genetic diversity was detected in Cymbidium goeringii for both population level (P = 63.1%; He = 0.194 5) and species level (P = 88.2%; He = 0.262 8). A higher level of genetic differentiation was detected among populations (G _ST = 0.244 0, F _ST = 0.220 7) with Nei’s G _ST analysis and analysis of molecular variance (AMOVA), and no correlation was found between geographical and genetic distance. Genetic drift rather than gene flow played an important role in forming the present population structure of Cymbidium goeringii. Limited gene flow among populations and gene drift increase the extinction risk of local populations. Some conservation concerns are therefore discussed together with possible strategies for implementing in situ and ex situ conservation. __________ Translated from Biodiversity Science, 2006, 14(3): 250–257 [译自: 生物多样性] Equally contributed authors     

春兰CgSEP3基因的克隆和表达分析

该研究采用RT-PCR和RACE技术从春兰(Cymbidium goeringii)中分离到1个SEPALLATA3(SEP3)基因。序列分析表明,该基因含有1个732bp的开放阅读框(ORF),共编码243个氨基酸。系统进化树分析显示,该基因是MADS-box基因家族AP1/AGL9组SEP的同源基因,其编码蛋白与其它植物SEP3类蛋白具有较高的一致性,命名为CgSEP3(登录号为KF924272)。实时荧光定量分析表明,CgSEP3在春兰花器官中均有表达,其中在唇瓣、侧瓣和萼片中的表达量较高,在子房和蕊柱中的表达量较低;而且CgSEP3在花发育各个时期都有表达,在1~2cm的花芽中表达量最高,在盛开的花中的表达量最低。研究认为,CgSEP3基因可能在春兰花瓣和萼片的形成过程中具有重要作用。     

春兰AGL6-3基因的克隆及实时定量表达分析

该研究以春兰(Cymbidium goeringii)正常花及其2枚侧瓣突变成唇瓣样的花瓣(简称:蝶花)为实验材料,采用RT-PCR结合RACE技术从春兰中分离出AGL6-3基因。序列分析表明,AGL6-3基因在春兰正常花和蝶花中序列相同,该基因含有1个720bp长的开放阅读框(ORF),共编码239个氨基酸。系统进化树进行分析表明,该基因属于MADS-box基因中AP1/AGL9组的AGL6同源基因,命名为CgAGL6-3(基因登录号为KU058679)。实时荧光定量表达分析表明,CgAGL6-3在春兰正常花和蝶花各花器官中表达存在差异。在正常春兰中CgAGL6-3基因在唇瓣中强烈表达,在主萼、侧萼及蕊柱中表达量较低,在侧瓣中则微乎其微;而在蝶花中CgAGL6-3基因在唇瓣中强表达,侧瓣中的表达量次之,在主萼、侧萼和蕊柱中的表达量相近且均较低。研究说明,CgAGL6-3基因可能在春兰蝶花侧瓣唇瓣化的过程中扮演重要角色。     

气候变暖背景下春兰和蕙兰的适生区分布预测

为了阐明气候变暖背景下春兰(Cymbidium goeringii)和蕙兰(C. faberi)在我国的适生区分布变化情况,根据157条分布记录和19个生物气候变量,应用最大熵物种分布模型,对2070年4种温室气体排放情景下春兰和蕙兰在我国的适生区分布进行预测,并筛选影响其地理分布的主要气候因子。结果表明:(1)2070年春兰和蕙兰分布点的年均温(bio1)、最冷月最低温度(bio6)和最冷季平均温度(bio11)等均升高,气候有变暖趋势;(2)受试者工作特征曲线下面积(AUC)值在0.9—1.0之间,模型预测结果可信度较高;(3)影响春兰、蕙兰当前和2070年地理分布的限制性气候因子主要有最冷月最低温度(bio6)、最冷季平均温度(bio11)、年均降水量(bio12)和最干月份降水量(bio14);(4)气候变暖将会对春兰和蕙兰的适宜生境范围和面积产生影响。预测2070年春兰的适宜生境面积将会有所减小,而蕙兰的适宜生境面积将会增加,且整体有向北迁移的趋势。研究结果为野生春兰和蕙兰的生态风险评价和引种提供了重要依据。     

Mycorrhizas alter nitrogen acquisition by the terrestrial orchid Cymbidium goeringii

Background and Aims

Orchid mycorrhizas exhibit a unique type of mycorrhizal symbiosis that occurs between fungi and plants of the family Orchidaceae. In general, the roots of orchids are typically coarse compared with those of other plant species, leading to a considerably low surface area to volume ratio. As a result, orchids are often ill-adapted for direct nutrient acquisition from the soil and so mycorrhizal assocaitions are important. However, the role of the fungal partners in the acquisition of inorganic and organic N by terrestrial orchids has yet to be clarified.

Methods

Inorganic and amino acid N uptake by non-mycorrhizal and mycorrhizal Cymbidium goeringii seedlings, which were grown in pots in a greenhouse, was investigated using a ¹⁵N-labelling technique in which the tracer was injected at two different soil depths, 2·5 cm or 7·5 cm. Mycorrhizal C. goeringii seedlings were obtained by inoculation with three different mycorrhizal strains isolated from the roots of wild terrestrial orchids (two C. goeringii and one C. sinense).

Key Results

Non-mycorrhizal C. goeringii primarily took up NO₃⁻ from tracers injected at 2·5-cm soil depth, whereas C. goeringii inoculated with all three mycorrhiza primarily took up NH₄⁺ injected at the same depth. Inoculation of the mycorrhizal strain MLX102 (isolated from adult C. sinense) on C. goeringii roots only significantly increased the below-ground biomass of the C. goeringii; however, it enhanced ¹⁵NH₄⁺ uptake by C. goeringii at 2·5-cm soil depth. Compared to the uptake of tracers injected at 2·5-cm soil depth, the MLX102 fungal strain strongly enhanced glycine-N uptake by C. goeringii from tracers injected at 7·5-cm soil depth. Cymbidium goeringii inoculated with CLB113 and MLX102 fungal strains demonstrated a similar N uptake pattern to tracers injected at 2·5-cm soil depth.

Conclusions

These findings demonstrate that mycorrhizal fungi are able to switch the primary N source uptake of a terrestrial orchid, in this case C. goeringii, from NO₃⁻ to NH₄⁺. The reasons for variation in N uptake in the different soil layers may be due to possible differentiation in the mycorrhizal hyphae of the C. goeringii fungal partner.     

Analysis of diversity and relationships among Chinese orchid cultivars using EST-SSR markers

Chinese orchid (Cymbidium spp.) is an important potted flower with extremely high ornamental value in China. It is important to identify the genetic diversity of its germ plasma resources for development and evaluation of its cultivars. A set of 13 expressed sequence tag (EST)-derived simple sequence repeat (SSR) was used to analyze 103 cultivars of six species of Chinese orchid, namely Cymbidium goeringii, C. faberi, Cymbidium ensifolium, C. kanran, C. sinense and C. goeringii var. longibracteatum. The 13 SSR primer pairs generated a total of 168 polymorphic bands, with an average of 12.92 bands per primer and a range of 6–24 bands which clearly revealed the difference between cultivars inter- or intra-species of Chinese orchid. Cluster analysis based on UPGMA, NJ and PCoA method showed a dendrogram with three basic clusters and splitting feature of C. ensifolium and C. goeringii which partially congruent with the current taxonomic classification.     

Molecular diversity and relationships among <Emphasis Type="Italic">Cymbidium goeringii</Emphasis> cultivars based on inter-simple sequence repeat (ISSR) markers

Spring orchid (Cymbidium goeringii) is a popular flowering plant species. There have been few molecular studies of the genetic diversity and conservation genetics on this species. An assessment of the level of genetic diversity in cultivated spring orchid would facilitate development of the future germplasm conservation for cultivar improvement. In the present study, DNA markers of intersimple sequence repeats (ISSR) were identified and the ISSR fingerprinting technique was used to evaluate genetic diversity in C. goeringii cultivars. Twenty-five ISSR primers were selected to produce a total of 224 ISSR loci for evaluation of the genetic diversity. A wide genetic variation was found in the 50 tested cultivars with Nei’s gene diversity (H = 0.2241) and 93.75% of polymorphic loci. Fifty cultivars were unequivocally distinguished based on ISSR fingerprinting. Cultivar-specific ISSR markers were identified in seven of 50 tested cultivars. Unweighted pair-group mean analysis (UPGMA) and principal coordinates analysis (PCA) grouped them into two clusters: one composed the cultivars mainly from Japan, and the other contained three major subclusters mainly from China. Two Chinese subclusters were generally consistent with horticultural classification, and the third Chinese subcluster contained cultivars from various horticultural groups. Our results suggest that the ISSR technique provides a powerful tool for cultivar identification and establishment of genetic relationships of cultivars in C. goeringii.     

云南野生春兰根系内生微生物多样性的分析

春兰(Cymbidium goeringii)是一种具有较高经济和观赏价值的地生兰。由于栖息地的严重破坏,大多数地生兰处于濒危状态,根系与微生物的共生关系伴随着兰科植物从种子萌发到开花结果,因此根系内生微生物在地生兰生活史中具有重要作用。【目的】分析野生春兰根系内生菌群落组成与潜在功能,为春兰的人工保育提供理论依据。【方法】采集云南省昆明市、保山市和大理州的野生春兰的根系,利用宏基因组测序,并进行物种注释及功能注释。【结果】保山的春兰根内微生物丰度及多样性大于大理及昆明的春兰样品,春兰根系内生真菌的优势门为担子菌门(Basidiomycota)、子囊菌门(Ascomycota)和球囊霉门(Glomeromycota),优势科为球囊霉科(Glomeraceae)、多孔菌科(Polyporaceae)和角担菌科(Ceratobasidiaceae),其中角担菌科是春兰主要的兰科菌根真菌(orchid mycorrhizal fungi, OMF);内生细菌的优势门为厚壁菌门(Firmicutes)、变形菌门(Proteobacteria)和拟杆菌门(Bacteroidetes),优势科为韦...     

High photosynthetic photon flux and high CO2 concentration under increased number of air exchanges promote growth and photosynthesis of four kinds of orchid plantlets in vitro

Summary In vitro plantlets of Phalaenopsis ‘Happy Valentine’, Neofinetia falcate Hu, Cymbidium kanran Makino, and Cymbidium goeringii Reichb. f. were grown under photoautotrophic [high photosynthetic photon flux (PPF), high CO₂ concentration, and increased number of air exchanges] and heterotrophic (low PPF, low CO₂ concentration, no air exchanges) culture conditions. After 40 d of culture, a significant difference in plantlet growth was observed between the two cultures. Total fresh and dry mass were on average 1.5 times greater in photoautotrophic culture than in heterotrophic culture. Higher net photosynthetic rates were also observed for Phalaenopsis in photoautotrophic culture. In photoautotrophic culture, little difference was observed in air temperature between the inside and outside of the culture vessel, whereas in heterotrophic culture, air temperature inside the culture vessel was 1–2°C higher than that outside the culture vessel. Relative humidity inside the culture vessel was remarkably different between the two cultures: 83–85% in photoautotrophic culture and 97–99% in heterotrophic culture. These results indicated that growth and net photosynthetic rate of in vitro orchid plantlets were susceptible to the culture environments such as PPF, CO₂ concentration, relative humidity (RH), and the number of air exchanges, which would allow a more efficient micropropagation system for these orchid plants.