人工种植天麻的影响因子优化研究

天麻是一种名贵的中草药,可用于治疗多种疾病。自然条件下,天麻通过与真菌蜜环菌的共生从腐木获取营养进行生长。为了对人工大规模种植天麻的影响因子进行优化研究,利用正交设计方法,在昭通小草坝地区开展了一个样本量较大的种植实验。在测试的4种因子中,接种蜜环菌对种植天麻影响最大：其次,在露天灌木丛地种植的天麻产量明显高于在林地下的种植。此外,开展了12个树种对蜜环菌生长以及天麻种植的影响研究。结果发现十齿花（Dipentodonsinicus）对蜜环菌的生长最好,同时,也在天麻种植实验中有最高的产量;然而,添加磷、钾元素并不提高天麻的产量。     

不同菌材对天麻(Gastrodia elata)产量的影响

以速生树种亮叶桦（Betula luminifera Winkler）和响叶杨（Populus adenopoda Maxim）、经济树种核桃（Juglans regia Linn）为菌材,壳斗科菌材白栎（Quercus fabri Hance）为对照,分别设置不同的菌材用量和麻种用量水平,仿野生栽培红杆天麻（Gastrodia elata Bl.f.elat）和乌杆天麻（Gastrodia elata Bl.f.glauca S.Chow）,分析不同菌材对天麻产量的影响,筛选出高产的菌材种类、用量和天麻变型及麻种用量的组合,探讨亮叶桦、响叶杨和核桃代替壳斗科植物栽培天麻的可行性。结果表明：不同菌材对红杆天麻和乌杆天麻产量的影响有差异。两种天麻变型对菌材的偏好不同,红杆天麻在白栎和亮叶桦菌材处理下的平均产量高于响叶杨和核桃菌材处理下的产量,而乌杆天麻在亮叶桦和核桃菌材处理下的产量高于响叶杨和白栎菌材处理下的产量。两种天麻变型的最高产量的组合均出现在亮叶桦菌材伴栽的处理中。响叶杨伴栽天麻,效果不及亮叶桦和核桃。4种菌材对天麻的折干率影响不明显。在实际生产中,以亮叶桦、核桃和响叶杨作为替代菌材是完全可行的。从72个处理中筛选出高产的组合,单位面积菌材和麻种用量分别如下：12 kg亮叶桦菌材搭配0.8 kg红杆天麻麻种,12 kg核桃菌材搭配0.6 kg红杆天麻麻种,14 kg响叶杨菌材搭配0.8 kg红杆天麻麻种;14 kg亮叶桦菌材搭配0.6 kg乌杆天麻麻种,12 kg亮叶桦菌材搭配0.6 kg乌杆天麻麻种,10 kg核桃菌材搭配0.8 kg乌杆天麻麻种。     

不同菌材对天麻(Gastrodia elata)产量的影响

以速生树种亮叶桦(Betula luminifera Winkler)和响叶杨(Populus adenopoda Maxim)、经济树种核桃(Juglans regia Linn)为菌材,壳斗科菌材白栎(Quercus fabri Hance)为对照,分别设置不同的菌材用量和麻种用量水平,仿野生栽培红杆天麻(Gastrodia elata Bl.f.elat)和乌杆天麻(Gastrodia elata Bl.f.glauca S.Chow),分析不同菌材对天麻产量的影响,筛选出高产的菌材种类、用量和天麻变型及麻种用量的组合,探讨亮叶桦、响叶杨和核桃代替壳斗科植物栽培天麻的可行性。结果表明:不同菌材对红杆天麻和乌杆天麻产量的影响有差异。两种天麻变型对菌材的偏好不同,红杆天麻在白栎和亮叶桦菌材处理下的平均产量高于响叶杨和核桃菌材处理下的产量,而乌杆天麻在亮叶桦和核桃菌材处理下的产量高于响叶杨和白栎菌材处理下的产量。两种天麻变型的最高产量的组合均出现在亮叶桦菌材伴栽的处理中。响叶杨伴栽天麻,效果不及亮叶桦和核桃。4种菌材对天麻的折干率影响不明显。在实际生产中,以亮叶桦、核桃和响叶杨作为替代菌材是完全可行的。从72个处理中筛选出高产的组合,单位面积菌材和麻种用量分别如下:12 kg亮叶桦菌材搭配0.8 kg红杆天麻麻种,12 kg核桃菌材搭配0.6 kg红杆天麻麻种,14 kg响叶杨菌材搭配0.8 kg红杆天麻麻种;14 kg亮叶桦菌材搭配0.6 kg乌杆天麻麻种,12 kg亮叶桦菌材搭配0.6 kg乌杆天麻麻种,10 kg核桃菌材搭配0.8 kg乌杆天麻麻种。     

天麻生长过程中可溶性蛋白和多糖的累积规律

以不同来源的蜜环菌 [(Armillariamellea(Vahl.exFr.)Qu啨ｌ.) ]4个菌株分别伴栽红天麻 (GastrodiaelataBl.f.elata)、乌天麻 (GastrodiaelataBl.f.glaucaS .Chow)以及红乌杂交天麻 ,对天麻生长发育过程中可溶性蛋白和多糖含量的变化规律进行了初步研究。结果表明 ,随着天麻的生长 ,菌株M1 、M2 、M3和M4 伴栽的红天麻、乌天麻以及红乌杂交天麻中 ,其可溶性蛋白含量均在逐步增加 ;在同一生长期内 ,4个菌株伴栽的红乌杂交天麻中可溶性蛋白含量大于红天麻和乌天麻。由菌株M1 、M2 、M3和M4 伴栽的红天麻、乌天麻以及红乌杂交天麻 ,其多糖含量呈现先升高后下降的趋势 ;在同一生长期中 ,4个菌株伴栽的红天麻的多糖含量大于乌天麻和红乌杂交天麻。     

The Structure and Antifungal Functions of Vegetative Propagation Corm of Gastrodia elata

The growth of Gastrodia elata Bl. and Armillaria mellea (Vahl. ex Fr.) Quel. shares a special symbiotic relation. In general, A. mellea invades the G. elata , the epidermal cells, the cortical cells and the large cells of the growing vegetative propagation corm of G. elata . The empty cavity cells, the cork cells of the isolation in the vegetative propagation corms and the large cells of G. elata were the defensive structure, protecting the new G. elata from pathological invasion by A. mellea . In winter, G. elata enters the stage of hibernation.　The faulting layer derived from the cork cells of the isolation was the last defensive structure by which new G. elata could safely live through the winter.     

四株昭通乌天麻共生蜜环菌的生理生化特征及其分子鉴定

目的对4株昭通乌天麻(Gastrodia elata)共生蜜环菌(Armillaria mella)的生理生化活性进行研究,并鉴定其分类地位。方法通过测定不同碳源和氮源对蜜环菌生长以及胞外酶活性的影响,掌握该蜜环菌生理生化特性,再结合rDNA-IGS序列分析方法进行鉴定。结果不同蜜环菌需要不同的碳源、氮源。3种胞外酶中木聚糖酶活性最大,其次是羧甲基纤维素酶,漆酶活性最小。供试蜜环菌与云南、贵州地区天麻共生蜜环菌以及猪苓共生蜜环菌的亲缘关系比较远。结论蜜环菌能否作为天麻共生菌与其分类地位无关。     

The Relation Between Growth of Gastrodia elata Protocorms and Fungi

Armillaria mellea penetrated protocorms from seed germination and vegetative multiplication corms of Gastrodia elata with rhizomorph. At beginning, they formed a hypha passing road and a hypha flow in the inner cells of cortex, and then, they both penetrated inside of large cells and penetrated outside of cortical cells. Gastrodia elata seeds depended on digesting Mycenct osmundicola etc gain nutrition to germinate at the stage of sexual reproduction, but its corms of vegetative multiplication must be penetrated by Armillaria mellea obtaining nutrition for normal growth at the stage of vegetative propagation.     

天麻球茎不同部位的细胞对蜜环菌侵染的反应

用电镜技术比较研究了天麻球茎皮层细胞和中柱细胞对蜜环菌侵染的反应,皮层细胞在蜜环菌菌丝的刺渺下能产生囊状结构,然后这些小囊将入侵的菌丝包围和消化,而中柱细胞则不能,上述两种细胞在功能上的主要差别就在于此。     

密环菌胞外多糖的发酵条件研究

密环菌是天麻的共生菌,有研究证明,密环菌的菌丝及发酵液有与天麻相类似的药理作用,为此,我们进行密环菌深层发酵产胞外多糖条件的研究。研究结果表明：菌丝生长和多糖产生的最适初始pH为5．9左右;消泡剂用量在一定内增加,有利于菌丝的生长,但增加消泡剂用量会不利于多糖的产生;接种量大,菌丝得率高,但接种量为10％时的多糖得率最高;装液量试验表明,通气量大,有利于菌丝的生长和多糖的产生;机械搅拌对菌丝的生长和多糖的产生都不利;生长动态实验证明,密环菌可在6d内完成液体发酵;以豆粕粉作氮源时,菌丝得率最高,以麸皮作氮源时,多糖产量最高;红薯粉为密环菌菌丝生长和多糖产生的最适碳源。     

天麻中抗真菌蛋白质的诱导和积累

前曾报告从天麻（Ｇａｔｒｏｄｉａ ｅｌａｔａ Ｂｌ）中分离出一种对木霉丝生长有强抑制作用的蛋白质,命名为天麻抗真菌蛋白,简称ＧＡＦＰ,也称为Ｇａｓｔｒｏｄｉａｎｉｎ。对不同天麻材料来源的ＧＡＦＰ分析表明：ＧＡＦＰ的相对分量为１４ｋＤ,等电点可能不同,变动范围８．１－９．３。体外抑菌试验证明ＧＡＦＰ对腐生性真菌如木霉如木霉和密环菌等有强抑制活性,半抑制浓度ＩＣ０．５为０．０８ｍｇ／ｍＬ;对寄生免疫荧     

Nitrogen source effects on rhizosphere pH and nutrient accumulation by Pacific Northwest conifers

Growth responses ofCasuarina cunninghamiana to inoculation withFrankia are described in unsterilized field soils at three sites. At Mt Crawford, South Australia, seedlings of three provenances ofC. cunninghamiana were inoculated with a singleFrankia source just prior to planting out. Forty-four months after planting, inoculation had more than doubled wood production by twoC. cunninghamiana provenances, whilst a third provenance grew poorly and did not respond to inoculation. In Zimbabwe, seedlings of one provenance ofC. cunninghamiana were inoculated in the nursery with one of four differentFrankia strains. In an N deficient soil at Kadoma, three of theseFrankia increased tree height 14 months after planting by between 50% and 70% in comparison to the uninoculated seedlings. The fourthFrankia strain resulted in increased tree height to three times that of the uninoculated controls and up to double that of the other threeFrankia strains. At Gympie, Queensland, Australia, seedlings ofC. cunninghamiana raised open-rooted in a nursery bed were inoculated withFrankia seventeen weeks before planting out. During the 22 months following planting in the field, tree growth was limited by soil P status and there was no response in tree height or stem diameter to inoculation withFrankia or to N fertilizer unless P was applied. In the presence of added P there was a significant response both toFrankia inoculation and to N fertilizer. This positive interaction between P application and N treatment was reflected in wood volumes-inoculated trees and those trees supplied N fertilizer produced 34% and 95% more wood volume than did the uninoculated trees. These results demonstrate the potential to increase the productivity of Casuarina plantings by inoculation withFrankia and by alleviation of P deficiency.     

Responses of barley,pea, and maize to inoculation with different vesicular-arbuscular mycorrhizal fungi in irradiated soil

Summary The efficiency of different VAM fungi was investigated by inoculating barley, pea, and maize with different VAM fungi in irradiated soil in pots buried in the field. VAM frequency, growth and nutrient uptake were measured. In barleyGlomus epigaeus (CA) andG. macrocarpus (CA) were the most efficient out of 11 tested species and increased yield of grain by 24% and 21%, though they were not significant according to oneway analysis of variance. In pea, yield of grain was significantly increased from 46% to 104% (mean=68%) by 7 out of 10 tested species and by 105% by application of P fertilizer. The most efficient species wereG. epigaeus (CA),G. mosseae (GB), andG. etunicatus (CA). In maizeG. mosseae (GB) andG. caledonius (DK) increased total yield significantly by 59% and 47% in one experiment and in another experiment yield of cob was increased by 68% byG. mosseae (GB), 72% byG. caledonius (DK), and by 153% by application of P fertilizer. This experiment demonstrated that responsiveness to inoculation by VAM fungi differed among plant species, and that efficiency of different VAM fungi differed.     

Phylogenetically diverse AM fungi from Ecuador strongly improve seedling growth of native potential crop trees

In many deforested regions of the tropics, afforestation with native tree species could valorize a growing reservoir of degraded, previously overused and abandoned land. The inoculation of tropical tree seedlings with arbuscular mycorrhizal fungi (AM fungi) can improve tree growth and viability, but efficiency may depend on plant and AM fungal genotype. To study such effects, seven phylogenetically diverse AM fungi, native to Ecuador, from seven genera and a non-native AM fungus (Rhizophagus irregularis DAOM197198) were used to inoculate the tropical potential crop tree (PCT) species Handroanthus chrysanthus (synonym Tabebuia chrysantha), Cedrela montana, and Heliocarpus americanus. Twenty-four plant-fungus combinations were studied in five different fertilization and AMF inoculation treatments. Numerous plant growth parameters and mycorrhizal root colonization were assessed. The inoculation with any of the tested AM fungi improved seedling growth significantly and in most cases reduced plant mortality. Plants produced up to threefold higher biomass, when compared to the standard nursery practice. AM fungal inoculation alone or in combination with low fertilization both outperformed full fertilization in terms of plant growth promotion. Interestingly, root colonization levels for individual fungi strongly depended on the host tree species, but surprisingly the colonization strength did not correlate with plant growth promotion. The combination of AM fungal inoculation with a low dosage of slow release fertilizer improved PCT seedling performance strongest, but also AM fungal treatments without any fertilization were highly efficient. The AM fungi tested are promising candidates to improve management practices in tropical tree seedling production.     

不同海拔两种天麻仿野生栽培下产量和品质变化

为促进黔西北天麻的稳产高产以及山区经济的发展,本研究采用完全试验设计的方法,对天麻（Gastrodiaelata Blume）进行仿野生有性繁殖试验,探讨不同海拔下2个天麻品种在产量和品质上的变化,以及栽培天麻的经济效益分析。结果表明,在产量上,海拔1400～1800 m范围内,红、绿天麻以1800 m处的产量最高,其中红天麻的箭麻产量（2836.80 g/m2）、麻种产量（3799.90 g/m2）和箭麻数（38.33个/m2）均显著高于绿天麻;在品质上,红、绿天麻以1800 m处品质最优,其中红天麻在折干率（16.33%）上显著低于绿天麻（19.25%）,而在天麻素（4.43‰）和天麻多糖含量（36.63%）上则高于绿天麻（4.26‰,31.10%）。通过对不同海拔下2种天麻的经济效益分析可知,对黔西北地区来说,在海拔1800 m处栽培红天麻能获得较高的经济效益。     

The Biological Relationship of Gastrodia elata and Armillaria Melle

We have done a series of experiments on the biological relationship of G. elata and Armillaria mellea (Vahl ex Fr.) Quel on the basis of popular experience. The results obtained are as follows: 1. The whole ontogenetic pattern of G. elata may be generally divided into four stages: seedling formation, tuber formation, flowering and fruiting. Except for the flowering and fruiting stage, the activity of Amella determines the formation of G. clara seedlings, the renewal of its vegetative organs, and transformation of vegetative growth to reproductive growth. The germination of tile dormant buds and their development are also decided by the sites and the influence of infection of A mellea . In some cases, when the mixed bud of G. elara was injured, it could also form several tubers with the infection of A. mellea . 2. The infecting process of A. mellea on G. elata has been studied in detail. Some of our observations are not well fitted with Kusano's experimental results. We find that there are two modes of infection of A. mellea on G. clara: physiological infection and pathological infection. Under normal environmental conditions, A. mellea infects the tissues of cortical layer only, and may also lead to the enlargment and malformation of nuclei. On the other hand; the digestive cells possess both the functions of defense and infecting hyphae. The pathological infection often exists under unfavorable conditions. 3. The tuber of G. data is not the only organ to assimilate the A. mellea, the vegetative stem also has a function of assimilation. 4. The growihg tuber would not become an assimilating organ until winter was over.     

Effects of Rhodobacter capsulatus inoculation in combination with graded levels of nitrogen fertilizer on growth and yield of rice in pots and lysimeter experiments

A pot and a lysimeter experiment were carried out to study the effects of inoculation of the roots of rice seedlings with R. capsulatus in combination with graded levels of nitrogen (N) fertilizer on growth and yield of the rice variety Giza 176. Inoculation increased all the measured growth parameters and yield attributes, but the statistically significant differences at all N levels tested were only those for plant dry weight, number of productive tillers, grain and straw yields. The absolute increases in grain yield of the pot experiment due to inoculation were 0.63, 0.93 and 1.22 ton ha^–1 at 0, 47.6 and 95.2 kg N ha^–1, respectively. The results suggest that inoculation along with 47.6 kg N ha^–1 can save 50% of the nitrogen fertilizer needed for optimum G176 rice crop. However, inoculation along with 95.2 kg N ha^–1 can increase grain yield by about 1.2 ton ha^–1. This is probably the first reported evidence of a beneficial effect of phototrophic purple nonsulphur bacteria on rice growth and yield under flooded soil conditions.     

Studies on the Character istic of the Structures in the Cortical Cells of Gastrodia clara after Infection of Armillaria mellea

Aided by the techniques of thin and ultrathin sectioning and electron microscopy, the characteristic of structures in the cortical cells of Gastrodia elata was further investigated after infection of Armillaria mellea. It was found that the “hyphal coils”, observed with light microscope, in the cortical cells of G. elata were saccate structures deriv- ed from the cytoplasm of cortical cells and enclosed hyphae. And the cell walls of hyphae were digested in these sacs. Then, these hyphae without cell wall were cut into protoplast fragments in inner-most cortical cells. The results indicated that the cortical cells of G. elata possess digestive function.     

Displacement of an herbaceous plant species community by mycorrhizal and non-mycorrhizal Gmelina arborea, an exotic tree, grown in a microcosm experiment

Gmelina arborea Roxb. (Gmelina, Yemane) is a fast growing tree, native from India and considered as a potentially invasive woody plant in West Africa. Mycorrhizal inoculation of seedlings with Glomus intraradices was performed to study (1) the effect on the growth of G. arborea, (2) the impact on the catabolic diversity of soil microbial communities and (3) the influence on the structure of herbaceous plant species communities in microcosms. Treatments consisted of control plants, pre-planting fertilizer application and arbuscular mycorrhizal (AM) inoculation. After 4 months’ culture in autoclaved soil, G. arborea seedlings were either harvested for growth measurement or transferred into containers filled with the same soil but not sterilized. Other containers were kept without G. arborea seedlings. After 12 months’ further culture, effects of fertilizer amendment and AM inoculation on the growth of G. arborea seedlings were recorded. AM colonization was significantly and positively correlated with plant diversity. The substrate-induced respiration response to carboxylic acids was significantly higher in the absence of G. arborea and in the presence of G. intraradices as compared to the other treatments. The influence of AM symbiosis on plant coexistence and on allelopathic processes of invasive plants are discussed.     

A multiplex PCR-based method for the detection and early identification of wood rotting fungi in standing trees

AIMS: The goal of this research was the development of a PCR-based assay to identify important decay fungi from wood of hardwood tree species in northern temperate regions. METHODS AND RESULTS: Eleven taxon-specific primers were designed for PCR amplification of either nuclear or mitochondrial ribosomal DNA regions of Armillaria spp., Ganoderma spp., Hericium spp., Hypoxylon thouarsianum var. thouarsianum, Inonotus/Phellinus-group, Laetiporus spp., Perenniporia fraxinea, Pleurotus spp., Schizophyllum spp., Stereum spp. and Trametes spp. Multiplex PCR reactions were developed and optimized to detect fungal DNA and identify each taxon with a sensitivity of at least 1 pg of target DNA in the template. This assay correctly identified the agents of decay in 82% of tested wood samples. CONCLUSIONS: The development and optimization of multiplex PCRs allowed for reliable identification of wood rotting fungi directly from wood. SIGNIFICANCE AND IMPACT OF THE STUDY: Early detection of wood decay fungi is crucial for assessment of tree stability in urban landscapes. Furthermore, this method may prove useful for prediction of the severity and the evolution of decay in standing trees.     

Dynamics of bioluminescence by Armillaria gallica, A. mellea and A. tabescens

Although fungal bioluminescence is well documented, the ecological significance is poorly understood. We examined bioluminescence by three sympatric species of Armillaria wood decay fungi, differing in parasitic ability. Luminescence by mycelia of four genets of A. gallica, A. mellea and A. tabescens was examined in response to environmental illumination or mechanical disturbance. Luminescence dynamics were assessed in a time series of measurements every 2 min for 72 h for mycelia growing on malt agar or on Cornus florida root wood. Luminescence by the necrotrophic species A. gallica was enhanced by environmental illumination and mechanical disturbance of mycelia. In contrast luminescence by the more parasitic A. mellea and A. tabescens was quenched by prolonged exposure to environmental illumination and less responsive to mechanical disturbance. With environmental illumination absent, all mycelia representing six genets of each Armillaria species were constitutively luminescent. The temporal dynamics of luminescence by all mycelia were complex with no evidence of the previously reported diurnal periodicity. Differences among Armillaria spp. in bioluminescence expression might reflect differences in ecological context as well.