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。     

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.     

Study of Factors for Cultivating the Orchid Species Gastrodia elata,a Traditional Chinese Medicine

The orchid speices Gastrodia elata is a valuable traditional Chinese medicine and has been widely applied for treating a variety of diseases. The yield of wild Gelata is very limited since its vegetative growth is exclusively dependent on its symbiosis with the fungus Armillaria mellea, from which Gelata is able to obtain nutrients from rotten wood in the forest. To develop a standard for cultivating Gelata in large quantities, four factors that may influence the yield of Gelata need to be further investigated, including cultivation environment, inoculation volume of Armillaria, wood from different tree species, and the effect of fertilizer on the maturation of Gelata. To optimize these factors, a large scale orthogonal experiment was performed in the farmland of Xiaocaoba, Zhaotong, Yunnan Province. Among the four factors tested, inoculation of Armillaria played the most important role in the maturation of Gelata. The yield of mature tubers, in terms of both tuber weight and quantity, on open ground is greater than that in forested areas. Of the 12 tree species tested, Dipentodon sinicus stimulated the largest amount of growth of Armillaria and produced the greatest yield of Gelata in the farmland. In comparison to the other factors tested, fertilizer showed no effect on the yield of G-elata.     

蜜环菌和天麻共生营养关系的放射性自显影研究

本文利用3H-葡萄糖以打孔浇灌法标记天麻。用标记的天麻伴栽蜜环菌,追踪标记化合物是否存在蜜环菌中。显微放射自显影的结果表明：天麻自身能吸收标记物,同化的标记物其长距离运输靠天麻的纵向维管组织进行。天麻表皮外的蜜环菌菌索的光镜、电镜自显影显示,蜜环菌能从天麻中获取营养,蜜环菌和天麻之间存在营养物质的相互交流,菌麻之间存在着特殊的共生关系。     

天麻3种变型开花生物学特性的研究

在天麻开花时,对略阳红秆、绿秆、乌秆天麻3种变型的花果期,花茎、花被筒及子房的长度,果实、种子及胚的长度、宽度等进行了观察研究,结果发现：天麻的这3种变型在每个方面都有一定的差异,乌秆天麻花茎最高,果实和种子最长,开花、结果日期最晚;红秆天麻花序轴、每株结果数、果实重量和种子直径最大,开花、结果日期最早;绿秆天麻花、果期最短,而胚的长度和直径却最大。     

天麻种子／原球茎和营养繁殖茎被菌根真菌定殖后的细胞分化

天麻（Gastrodia elata Bl.)种子可与紫萁小菇（Mycena osmundicola Lange),兰小菇（M.orchidicola Fan et Guo)等一类小菇属真菌共生萌发形成原球茎。侵入种皮的菌丝集结在柄状细胞外周的胚柄残迹中,首先侵入胚的柄状细胞,然后自柄状细胞侵入其他原胚细胞。原胚细胞发生功能分化,形成菌丝结细胞和消化细胞,初被菌丝定殖的原胚细胞具有消化菌丝的功能,随后,部分原胚细胞逐渐被菌丝充满,充育成菌丝结细胞。菌丝由菌丝结细胞进一步侵入消化细胞后最终被消化。由原球茎分化形成的营养繁殖茎（以下简称营繁茎）进一步被蜜环菌（Armilariella mellea(Vahl.Fr.)Karst.)定植,蜜环菌与紫箕小菇的菌丝同时存在于营繁茎中,但两菌相遇时都停止蔓延,互不交错侵染。     

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。     

Cytodifferentiation of the Seeds (Protocorms) and Vegetative Propagation Corms Colonized by Mycorrhizal Fungi

The stipecell, subepidermal parenchyma cells and inner cortical parenchyma cells were differentiated from Gastrodia elata Bl. seed and protocorm cells when they were colonized by the fungal hyphae of Mycena osmundicola Lange and M. orchidicola Fan et Guo. The hyphae aggregated in the suspensor remnant surrounding stipecell, primarily penetrated the stipecell, and then colonized the embryo of seed. Stipecell is the unique invading site of the hyphae. Subepidermal parenchyma cells containing pelotons of hyphae is also a kind of passage cells of hyphae, but, when primarily colonized by hyphae, they can degenerate a little of hyphae. The hyphae colonizing inner cortical parenchyma cells were totally degenerated, and the function of inner ocrtical parenchyma cells is digestive. The vegetative propagation corms, which differentiated from protocorms, were recolonized by Armilariella mellea (Vahl:Fr.) Karst., and the hyphae of A. mellea and M. osmundicola were found in the same cell, but there is a layer of cells uncolonized by mycorrhizal fungal hyphae. This means the two fungal species can not crisscross colonize the cell of G. elata.     

Exogenous Calcium Enhances the Formation of Vegetative Buds,Flowers and Roots in Tobacco Pith Explants Cultured in the Absence of Exogenous Hormones

Pith explants excised from the apical stem internodes of vegetative, flowering, and fruiting tobacco plants were cultured on hormone-free medium in the presence or absence of CaCl₂ (3 mM). The aim was to determine the role of exogenous calcium (Ca²⁺), applied at the concentration normally present in the Murashige and Skoog (1962) medium, in organ formation obtainable in the absence of the exogenous hormonal input. Exclusive formation of vegetative buds was obtained from explants excised from vegetative plants (pure vegetative programme); vegetative buds and flowers (and occasionally roots) on the same sample were obtained from explants from flowering plants (mixed flowering programme); whereas roots, very occasionally associated with vegetative buds and flowers on the same sample, were obtained from explants from fruiting plants (mixed rooting programme). Histological analysis showed that the organs always exhibited indirect regeneration. Exogenous Ca²⁺ promoted the formation of meristemoids and the first phases of their growth into organs, but did not change the realization of the organogenic programme and did not affect callogenesis. Instead, the influence of exogenous Ca²⁺ changed with the programme, when considering the last phases of organ growth (i.e., macroscopic development and elongation), and the appearance of morphological anomalies in the organs.     

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。     

天麻消化紫萁小菇及蜜环菌过程中细胞超微结构变化的研究

本文对天麻种子消化入侵的紫萁小菇菌丝及营养繁殖茎消化蜜环菌过程中,细胞超微结构的变化进行了研究。观察结果表明:紫萁小菇侵入种胚后,染菌胚细胞的细胞器逐渐消失,其细胞质产生囊状体起消化菌丝的作用,存在于胚细胞中的紫萁小菇菌丝有脱壁或失去细胞质成为空腔等变化;种子萌发形成的原球茎消化紫萁小菇的方式同胚萌发阶段类同。蜜环菌侵入原球茎分化的营养繁殖茎后,皮层细胞产生消化酶类颗粒或囊状体包围并分解蜜环菌菌丝;被皮层细胞消化的菌丝残物或部分菌丝进入皮层内面的大型细胞,此时大型细胞的代谢功能显著增强,该细胞中的各种水解酶颗粒及液泡等完成对菌体物质的最终同化。紫萁小菇及蜜环菌先后在天麻有性繁殖和无性繁殖阶段侵染供给其营养,但菌丝被消化过程中的细胞形态变化、被消化方式不完全一致。     

天麻矿物质吸收及其营养机理探讨

探讨了天麻吸收矿物质过程和天麻的营养机理。研究蜜环菌在天麻的金属元素、非金属元素吸收中起的作用, 阐明天麻的有机营养吸收过程。提出了天麻吸收非金属元素的两个途径,分析了天麻吸收金属元素的“泵”的作用机理。指出了天麻蜜环菌之间的游离氨基酸特别是天冬氨酸和谷氨酸的交流是天麻和蜜环菌生长发育的关键。     

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     

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。     

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.     

蜜环菌对镁的耐性和富集特性

研究了镁对蜜环菌生长的影响, 蜜环菌对于镁的耐性和富集规律, 以及高浓度镁胁迫下蜜环菌的抗氧化酶的变化情况。3?15 g/L的Mg浓度对于蜜环菌菌体的生长有促进作用。Mg浓度19 g/L以上时, 蜜环菌菌体的生长受到抑制。蜜环菌的子实体形成和子实体生物量在Mg的浓度为11 g/L以下时不受影响, 超过11 g/L则子实体不能萌发。皮壳状菌丝和菌索中Mg的含量随培养基中Mg浓度的增大而增大, 培养基Mg浓度达到16 g/L后, 菌丝、菌索中Mg的含量都不再上升。子实体对Mg的富集量比菌丝体小的多, 在培养基Mg浓度在9、10 g/L时, 子实体中Mg的含量与对照组有显著差异。随着培养基中Mg浓度的提高, 菌丝和菌索POD、CAT、SOD活性都有增加, 而且菌丝与菌索之间抗氧化酶活力的差异随着培养基中Mg浓度的提高而增大。     

Carbon source for reproduction in a spring ephemeral herb, Corydalis ambigua (Papaveraceae)

1.  The carbon source for reproduction in plants may differ between flowering and fruiting stages. To clarify how spring ephemerals use current photosynthetic products for reproduction, the allocation patterns of photosynthate at flowering and fruiting and the effects of resource limitation on reproductive performance in Corydalis ambigua were assessed.
2.  A ¹³C tracing experiment revealed that about 20% of the current photosynthetic carbon was used for reproduction at both flowering and fruiting. The proportion of ¹³C allocated to fruits was constant irrespective of the light level. In contrast, ¹³C translocation to tubers increased at fruiting, and this trend was accelerated when plants were shaded.
3.  Defoliation treatment significantly reduced nectar production and tuber mass, while seed production was not affected. Therefore, when carbon assimilation was limited, carbon was preferentially allocated to current reproduction (seeds) rather than to pollinator attraction (nectar) or storage (tuber).
4.  If seed production is partly supported by carbohydrate reserved in the old tissue of tubers, nectar and seed production may not compete strongly for carbon sources. In contrast to the ability of high seed production, the susceptibility of nectar production to current photosynthesis indicates that seed production of this species is basically limited by pollen capture.
5.  Therefore, temporal separation of resource pool for reproduction may mitigate the joint limitation of seed production between pollinator attraction and resource availability. Temporal variation of the sink–source balance of storage organ is crucial to understand the cost of reproduction in perennial plants.     

A STUDY 0N THE S0URCE OF SEC0NDARY NUTRIENTS F0R GASTRODIA ELATA BL.

Using isotopic labelling technique (with 32P. 15N. 86Rb), we demonstrated that Gastrodia elata Bl. might draw nutrients from the soil besides their main nutritional source Armillariella mellea (Vahl. ex Fr) Karst. Hence the soil may considered as its second nutritional source.The nutritional components in the soil(fertilizer)are able to enter Gartrodia elata directly or indirectly though the mediation of Armillariella mellea. They may influence the interrelationship between Gastrodia elata and Armillariella mellea, and the growth of the former. Therefore, the study of the influence of fertilization technique on the yield and quality of Gastrodia elata is of signifi- cance.