杏黄兜兰菌根研究与应用

采用根组织切片法从杏黄兜兰根内分离到13株真菌,镰刀菌和组丝核菌为优势菌。将组丝核菌JF74、JF75、JF80制成菌剂,施入杏黄兜兰根部,对杏黄兜兰产生了一定的促进生长的效果。     

带叶兜兰种子原地共生萌发及有效菌根真菌的分离与鉴定

为获得带叶兜兰(Paphiopedilum hirsutissimum)种子萌发的共生真菌,采用原地共生萌发技术获得了2株自然萌发的小幼苗,并分离和筛选出了有效的种子萌发共生菌——瘤菌根菌(Epulorhiza sp.)。为验证分离菌株对带叶兜兰种子萌发的有效性,将Phs34号菌株与带叶兜兰种子在灭菌后的原生境基质上进行室内共生萌发试验,结果表明,经过6周的培养,对照组没有观察到种子的萌发;接菌的种子胚明显膨大,突破种皮,形成原球茎,平均萌发率为(58.35±3.41)%。这表明分离得到的瘤菌根菌能促进带叶兜兰的种子萌发。     

基于灰色关联分析方法评价13种野生兜兰的迁地保护适应性

由于过度收集和对原生境地的破坏,我国野生兜兰已濒临灭绝,对其进行迁地保护十分必要。通过对引种的13种40份野生兜兰资源进行迁地栽培,并观测兜兰种质在迁移地的各种生长性状指标等,本研究利用灰色关联分析方法评价13种兜兰的迁地适应性,分析影响其迁地保护的关键因素。结果表明:(1)带叶兜兰、同色兜兰、长瓣兜兰、文山兜兰适应性较强,与理想种的关联度均在0.8以上;巨瓣兜兰、白花兜兰、小叶兜兰、紫纹兜兰、紫毛兜兰、硬叶兜兰适应性一般,与理想种的关联度在0.7~0.8之间;麻栗坡兜兰、杏黄兜兰、红旗兜兰适应性较差,与理想种的关联度均在0.6以下;(2)不同兜兰种类、同一种类不同产地兜兰种质的迁地生长适应性不同,来源地和栽培温度可能是影响其迁地保护的关键因子。     

兜兰属宽瓣亚属（广义）的被充研究

对新种小囊兜兰Paphiopedilum globulosum与新变种扁球兜兰P.micranthum var.oblatum作了描述与绘图。两者均为采自云南东南部的栽培植物,与硬叶兜兰P.micranthum近似。但小囊兜兰具有宽得多的叶（宽2．8－3．4cm),先端多少呈钩状、宽1．1－1．2cm的辱瓣,以及与唇瓣囊口几乎同样大小的退化雄蕊,而扁球兜兰则叶宽2．2－3cm,唇瓣扁球形,囊深不超过1cm,可区别于硬叶兜兰。上述种类均属于宽瓣亚属（广义）。该亚属有16个种产中国、越南或两地共有。本文提供了分种检索表。     

国产兜兰属植物观赏价值评价及其在华南地区的应用前景分析

以植株的生长状况、整株姿态、叶片观赏性、开花性、花形、花色和花朵观赏期为评价指标,利用灰色关联分析,对我国原产的27种兜兰属植物的观赏价值进行了综合评价并分析了它们在华南地区的应用前景。结果表明:多花长瓣型的飘带兜兰与理想种的关联度最高,达到0.7304,是最值得在华南地区栽培的兜兰种类,其次为白旗兜兰。而紫纹兜兰、汉氏兜兰;文山兜兰、格利兜兰、带叶兜兰、亨利兜兰、同色兜兰,它们的关联度系数超过0.6;紫毛兜兰、巨瓣兜兰、红旗兜兰、天伦兜兰、长瓣兜兰、根茎兜兰的关联度均超过0.5;也是值得推广兜兰种类;杏黄兜兰、硬叶兜兰、白花兜兰由于生长状况差、开花性差等原因与理想种的关联度分别只有0.1900、0.1921、0.2917,不适于华南地区栽培;其它9种兜兰的关联度虽然介于0.35~0.50之间,但其中有些种类如虎斑兜兰、麻栗坡兜兰、波瓣兜兰、德氏兜兰等由于具有独特的观赏价值和育种潜力,也是值得在华南地区栽培的兜兰种类。     

杏黄兜兰的花发育过程及引种栽培

通过形态解剖研究了杏黄兜兰花的发育过程,发现杏黄兜兰5月份发育出花序原基,6～7月份发育出花序鞘,7～8月份发育出苞片,8月底至9月完成花器官形态分化过程。形态分化完成后杏黄兜兰的花器官继续快速增长。退化雄蕊、能育雄蕊、花柱在花形成后早期不融合,在后期才融合形成合蕊柱,且因为相对生长速率的不同,三者的相对位置、形态也发生改变。此外,在7～9月份引种至昆明栽培的不同批次杏黄兜兰中,产生花芽比率显著不同,进一步证实8月底至9月初是杏黄兜兰进行花器官形态分化的关键时期。     

小叶兜兰的种子发育和无菌萌发

为建立小叶兜兰的繁育技术体系,本研究通过无菌播种的方法,辅以TTc生活力测定等方法,比较了小叶兜兰种子在授粉后不同发育时期和培养条件下的萌发率,对小叶兜兰种胚的发育过程进行了显微观察,探讨种胚发育程度与萌发的关系。结果表明,小叶兜兰种胚的发育阶段对萌发的影响最大,授粉后255d的种子萌发率最高（90．71％）,该阶段种子仍呈白色但微干燥,种胚刚发育至球形胚阶段,胚柄尚存。1／4MS和1／2MS为小叶兜兰适宜的基本培养基,添加100mg·L^-1的土豆汁对小叶兜兰的无菌萌发有良好的促进作用。     

同色兜兰的非共生萌发与快速繁殖研究

以授粉后不同发育时期的同色兜兰种子为材料,观察其形态特征和萌发过程,并探讨建立同色兜兰高效快繁体系的最佳条件。结果表明,种子发育中后期即授粉后210~240d为较适宜的采收期,授粉后210d的种子萌发率最高(达77.79%);1/4 MS和1/2MS为同色兜兰适宜的基本培养基,添加100mL/L椰乳或1g/L蛋白胨对种子萌发及原球茎生长和分化有明显的促进作用;添加1g/L活性炭对原球茎褐化有一定的抑制作用,但添加剂量不宜过大;添加香蕉汁和苹果汁对同色兜兰种子萌发和原球茎生长分化有抑制作用;暗处理对同色兜兰种子萌发无影响;分化后的原球茎在壮苗和生根培养基上培养120d即可得到4~5片叶、高3~5cm的同色兜兰健壮试管苗。     

兜兰属宽瓣亚属（广义）的补充研究

对新种小囊兜兰Paphiopedilum globulosum与新变种扁球兜兰P．micranthum var．oblatum作了描述与绘图。两者均为采自云南东南部的栽培植物,与硬叶兜兰P．micranthum近似。但小囊兜兰具有宽得多的叶(宽2．8～3．4cm),先端多少呈钩状、宽1．1～1．2cm的唇瓣,以及与唇瓣囊口几乎同样大小的退化雄蕊,而扁球兜兰则叶宽2．2～3 cm,唇瓣扁球形,囊深不超过1 cm,可区别于硬叶兜兰。上述种类均属于宽瓣亚属(广义）。该亚属有16个种产中国、越南或两地共有。本文提供了分种检索表。     

珍稀濒危植物硬叶兜兰的遗传多样性及遗传结构研究

由于人为采集、走私贩卖以及生境的破坏,分布于中国西南石灰岩地区的野生硬叶兜兰居群受到严重的干扰与威胁。为有效地保护这种珍稀野生植物,本研究采用ISSR和SRAP两种分子标记对15个硬叶兜兰野生居群进行遗传多样性及遗传结构的研究。结果表明,硬叶兜兰在物种水平上具有较高的遗传多样性（ISSR：PPB=91．66％,He=0．3839;SRAP：PPB=99．29％,Hc=0．2806）。硬叶兜兰居群间存在一定程度的遗传分化（ISSR：Gs1： 0．2577;SRAP：Gst=0．2383）,可能由于较低的基因流（ISSR：Nm=0．7201;SRAP：Nm=0．7991）所致。UPGMA聚类分析以及主成分分析均把15个居群分成2个主要分支。居群间的地理距离和海拔差距是引起居群遗传分化的自然因素。     

Interaction between Endophytic Fungi and Seedlings of Two Species of Paphiopedilum during Symbiotic Culture

Two fungi PM 1 and PA 1 were isolated from P.micranthum and P.armeniacum, respectively, the former belonged to Epulorhiza while the latter belonged to Tulasnella calospora. In this study, seedlings of P.armeniacum and P.micranthum were inoculated with PM 1 on DE medium, the biomass and the content of Ca, Mn of both P.armeniacum and P.micranthum seedlings were higher than those in the control seedlings, which indicated that PM 1 could improve the growth of those species. Seedlings of P.armeniacum and P.micranthum were inoculated with PA 1 on DE medium, the biomass in both species was no difference with those in the control ones. But PA 1 enhanced the dry weight of P.armeniacum according to the rate of dry weight increase. The impact of PM 1 on P.micranthum was higher than on P.armeniacum, while PA 1 impacted conversely, which demonstrated that fungi had stronger improving effect only on its own host plants instead of any other orchids. P.armeniacum seedlings wereinoculated with PM 1 and PA 1 on Harvais medium. The growth of seedlings was inhibited by fungi. The results indicated that nutrient level of medium played an important role in affecting the symbiotic relationship between P.armeniacum seedings and endophytic fungi. Poor nutrient medium was beneficial to the symbiotic relationship and nutrient rich medium was not conducive to the symbiosis.     

Effect of arbuscular mycorrhizal fungi inoculation of rice seedlings at the nursery stage upon performance in the paddy field and greenhouse

We examined the effect of arbuscular mycorrhizal fungi inoculation at the nursery stage on the growth and nutrient acquisition of wetland rice (t Oryza sativa L.) under field and pot conditions. Seedlings were grown on -ray sterilized paddy soil in two types of nurseries, namely dry nursery and wet nursery, with or without arbuscular mycorrhizal fungi (AMF) inoculation which was a mixture of indigenous AMF (t Glomus spp.) spores collected from the paddy field. Five-to-six week old seedlings were transplanted to the unsterilized soil under field and pot, respectively. Mycorrhizal seedlings had higher shoot biomass under both nursery conditions 5 weeks after sowing. Mycorrhizal colonization and sporulation were 2 to 3 times higher in the dry nursery than the wet nursery at the transplanting stage. Mycorrhizal colonization of plants inoculated in the nursery remained higher than those not inoculated under both field and pot conditions. Sporulation after transplanting to field conditions was about 10 times higher than in the pot. Inoculated plants produced higher biomass at maturity under field conditions, and the grain yield was 14-21% higher than those not inoculated. Conversely, grain yield and shoot biomass were not significantly influenced by AMF colonization under pot conditions. For plants originating from the dry nursery, N, P, Zn and Cu concentrations of field-grown plants at harvest were significantly increased by preinoculation with AMF over those left uninoculated. We conclude that the AMF inoculation at the nursery stage under both dry and wet conditions increased growth, grain yield and nutrient acquisition of wetland rice under field conditions.     

The effects of toxic metals,content of nutrients and inoculation with mycorrhizal fungi on the level of phenolics in roots and growth of Scots pine seedlings

The effects of two substrates and several strains of mycorrhizal fungi on the content of soluble phenolics in roots and growth of Scots pine seedlings was investigated. The first substrate was fertile and contaminated with copper, zinc and lead, whereas the second one displayed nutrient deficiency. The dry weights of needles, trunks, roots and the total biomass were higher in groups of seedlings inoculated with mycorrhizal fungi on fertile and polluted substrate. Inoculation of pine seedlings on this substrate resulted in a decrease in concentration of phenolics in roots and except for seedlings inoculated with Laccaria laccata negatively influenced the above-ground part: root ratio.     

Growth response of drought-stressed Pinus sylvestris seedlings to single- and multi-species inoculation with ectomycorrhizal fungi

Many trees species form symbiotic associations with ectomycorrhizal (ECM) fungi, which improve nutrient and water acquisition of their host. Until now it is unclear whether the species richness of ECM fungi is beneficial for tree seedling performance, be it during moist conditions or drought. We performed a pot experiment using Pinus sylvestris seedlings inoculated with four selected ECM fungi (Cenococcum geophilum, Paxillus involutus, Rhizopogon roseolus and Suillus granulatus) to investigate (i) whether these four ECM fungi, in monoculture or in species mixtures, affect growth of P. sylvestris seedlings, and (ii) whether this effect can be attributed to species number per se or to species identity. Two different watering regimes (moist vs. dry) were applied to examine the context-dependency of the results. Additionally, we assessed the activity of eight extracellular enzymes in the root tips. Shoot growth was enhanced in the presence of S. granulatus, but not by any other ECM fungal species. The positive effect of S. granulatus on shoot growth was more pronounced under moist (threefold increase) than under dry conditions (twofold increase), indicating that the investigated ECM fungi did not provide additional support during drought stress. The activity of secreted extracellular enzymes was higher in S. granulatus than in any other species. In conclusion, our findings suggest that ECM fungal species composition may affect seedling performance in terms of aboveground biomass.     

干旱胁迫条件下AMF促进小马鞍羊蹄甲幼苗生长的机理研究

采用温室水分控制试验,在干旱胁迫条件下,定量化研究优势丛枝菌根真菌(AMF)影响优势乡土植物小马鞍羊蹄甲(Bauhinia faberi var.microphylla)幼苗生长的机理,主要通过研究干旱胁迫条件下摩西球囊霉菌(Funneliformis mosseae)与小马鞍羊蹄甲的共生关系,阐明AMF在植物生长初期的作用。结果表明,干旱胁迫条件下,摩西球囊霉菌能够很好地侵染幼苗,侵染率高达89%—97%,并且不受水分条件影响。接种的幼苗最大光合速率、水分利用效率随着干旱胁迫程度从重度到轻度(水分从低到高)逐渐增大,相反地,叶片脯氨酸含量逐渐减小。接种显著地促进幼苗株高、叶片数、叶面积、根长、根面积等生长指标,提高幼苗各部分生物量、地上地下磷(P)含量。当含水量为60%田间持水量时,AMF促进小马鞍羊蹄甲幼苗吸收P的效果最好。接种还显著影响幼苗的生物量分配,在重度干旱胁迫时影响P分配,水分条件也显著影响幼苗的生物量分配。此外,接种和水分的交互作用对叶生物量、总生物量、生长指标以及地上部氮(N)总量影响显著。结果表明干旱胁迫条件下菌根效应显著,并在干旱条件下显著促进了小马鞍羊蹄甲幼苗的生长,这为进一步干旱河谷植被恢复提供了理论依据。     

丛枝菌根对喜树幼苗生长和氮、磷吸收的影响

喜树(Camptotheca acuminata)是我国特有的多年生亚热带落叶阔叶树种,因其次生代谢产物喜树碱具有良好的抗肿瘤活性而受到人们的广泛关注。该文通过温室盆栽接种试验,观察了2属6种丛枝菌根真菌即蜜色无梗囊霉(Acaulospora mellea)、光壁无梗囊霉(A. laevis)、木薯球囊霉(Glomus manihot)、地表球囊霉(G. versiforme)、幼套球囊霉(G. etunicatum)和透光球囊霉(G. diaphanum)对喜树幼苗生长和氮、磷养分吸收的影响。结果表明,丛枝菌根的形成对喜树幼苗的生长以及氮、磷营养的吸收均有影响。从生物量看,除幼套球囊霉和光壁无梗囊霉侵染形成的丛枝菌根喜树幼苗与无菌根幼苗(CK)差异不显著外,其余菌根幼苗的生物量均明显大于无菌根幼苗,透光球囊霉和蜜色无梗囊霉菌根幼苗尤为突出,分别达到无菌根幼苗的1.9和1.4倍。丛枝菌根的形成似乎不利于喜树幼苗的氮素营养吸收,并且主要体现在叶片的氮含量上。相反,丛枝菌根形成总体上促进喜树幼苗对磷素营养的吸收,并且主要体现在根的磷含量上。与无菌根幼苗比,所有菌根幼苗根的氮、磷分配比例增加,而叶片的氮、磷分配比例减少。     

共生真菌对铁皮石斛幼苗生理特性的影响

以铁皮石斛(Dendrobium catenatum)无菌组培苗为材料,接种C20、C22、C35、C42这4株真菌菌株.共生培养60d后,研究真菌对组培苗的平均鲜重增长率、营养元素、叶绿素和内源激素的影响.结果表明:(1)接菌组培苗的营养根重分离获得原接种菌株;(2)接菌组培苗的平均鲜重增长率,叶绿素a、b含量和营养元素N、P、K含量均高于对照;(3)接菌组培苗的GA3和IAA含量均高于对照,ABA含量低于对照.其中,接种C42的组培苗平均鲜重增长率和叶绿素a含量达极显著差异(p＜0.01),接种C22和C35组培苗的平均鲜重增长率和叶绿素a含量达到显著差异(p＜0.05),接种C20和C42的组培苗K含量达显著性差异,接种C20、C35、C42的组培苗IAA含量极显著地高于对照,接菌组培苗的N含量和GA3含量均达极显著差异,而接菌组培苗的ABA含量极显著地低于对照.因此,这4株真菌菌株可应用于铁皮石斛等兰科植物的栽培与生产中.     

Effects of mycorrhizal colonization on biomass production and nitrogen fixation of black locust (Robinia pseudoacacia) seedlings grown under elevated atmospheric carbon dioxide

Interactive effects of elevated atmospheric CO₂ and arbuscular mycorrhizal (AM) fungi on biomass production and N₂ fixation were investigated using black locust ( Robinia pseudoacacia ). Seedlings were grown in growth chambers maintained at either 350 μmol mol⁻¹ or 710 μmol mol⁻¹ CO₂. Seedlings were inoculated with Rhizobium spp. and were grown with or without AM fungi. The ¹⁵N isotope dilution method was used to determine N source partitioning between N₂ fixation and inorganic fertilizer uptake. Elevated atmospheric CO₂ significantly increased the percentage of fine roots that were colonized by AM fungi. Mycorrhizal seedlings grown under elevated CO₂ had the greatest overall plant biomass production, nodulation, N and P content, and root N absorption. Additionally, elevated CO₂ levels enhanced nodule and root mass production, as well as N₂ fixation rates, of non- mycorrhizal seedlings. However, the relative response of biomass production to CO₂ enrichment was greater in non-mycorrhizal seedlings than in mycorrhizal seedlings. This study provides strong evidence that arbuscular mycorrhizal fungi play an important role in the extent to which plant nutrition of symbiotic N₂-fixing tree species is affected by enriched atmospheric CO₂.     

Organic anion exudation by ectomycorrhizal fungi and Pinus sylvestris in response to nutrient deficiencies

Low molecular weight organic anions (LMWOA) can enhance weathering of mineral grains. We tested the hypothesis that ectomycorrhizal (EcM) fungi and tree seedlings increase their exudation of LMWOA when supply of magnesium, potassium and phosphorus is low to enhance the mobilization of Mg, K and P from mineral grains. Ectomycorrhizal fungi and Pinus sylvestris seedlings were cultured in symbiosis and in isolation on glass beads with nutrient solution or with sand as a rooting medium, with a complete nutrient supply or with Mg, K, P or N in low supply. Concentrations of all dicarboxylic LMWOA in the rooting medium were measured. Nonmycorrhizal seedlings released predominantly malonate. Colonization with Hebeloma longicaudum decreased the amount of organic anions exuded, whereas Paxillus involutus and Piloderma croceum increased the concentration of oxalate but not the total amount of LMWOA. Phosphorus deficiency increased the concentration of LMWOA by nonmycorrhizal and EcM seedlings. Magnesium deficiency increased the concentration of oxalate by nonmycorrhizal and EcM seedlings, but not the concentration of total LMWOA. Paxillus involutus grown in pure culture responded differently to low nutrient supply compared with symbiotic growth. Ectomycorrhizal fungi did not increase the total concentration of LMWOA compared with nonmycorrhizal seedlings but, depending on the fungal species, they affected the type of LMWOA found.     

辽东栎幼苗的外生菌根合成

 在温室花盆中播种辽东栎（Quercus liaotungensis）种子获得辽东栎幼苗,并对幼苗接种外生菌根真菌进行菌根合成试验。所用的外生菌根真菌有：铆钉菇（Gomphidius viscidus）、臭红菇（Russula foetens）、厚环乳牛肝菌（Suillus grevillei）、褐环乳牛肝菌（S. luteus）、彩色豆马勃（Pisolithus tinctorius）、美味牛肝菌（Boletus edulis）、赭丝膜菌（Cortinarius russus）、土生空团菌（Cenococ