植物组织培养方法生产药用次生代谢产物研究进展

许多药物来源于植物次生代谢途径,目前植物组织培养方法已成为生产药用成分的重要手段.在植物组织培养中,选择高产的外植体,寻找合适的培养条件,运用两相培养法和毛状根培养技术以及控制组织培养过程中的污染、褐化及玻璃化等问题,则是提高植物细胞生长速度和次生代谢产物产量并实现工业化生产的先决条件.本文主要从以上几个方面介绍其近来的研究进展,并提出了存在的问题及解决对策.     

高中新教材的组织培养实验

针对人教版的高中生物学新教材中加入了植物组织培养实验,介绍了一种简便的、适合高中生物学实验教学的菊花组织培养方法。详细介绍了铁盐母液的配制方法及配制和保存过程中的注意事项、各种激素的配制方法和培养基的配制中各个细节,从外植体的选取、清洗及接种各环节详细阐述了无菌培养物初代建立的关键技术。     

红豆杉组织培养及其产物紫杉醇研究进展

该文对红豆杉组织培养过程中外植体的选择、培养基、培养方法等方面研究进展进行了综述。获取抗癌药物紫杉醇主要途径有:红豆杉组织培养途径、人工种植途径、化学合成及微生物生产途径等。     

吊钟花的组织培养技术研究

以吊钟花(Enkianthus quinqueflorus Lour.)茎尖及带腋芽的茎段为外植体进行组织培养.结果表明,改良B5培养基(B5大量元素和钙盐+MS有机物、铁盐、微量元素)最有利于吊钟花的培养,外植体在改良B5+2,4D- 1 mg L-1的培养基中,愈伤组织诱导率可达100%.在含BA 1～2 m L-1+NAA0.1～0.5 mg L-1培养基中,可诱导产生不定芽.继代培养以改良B5+BA 1 mg L-1+NAA0.5 mg L-1培养基的增殖系数最高.生根培养基以1/2MS+IBA2 mg L-1为最佳,生根率可达80%以上.试管苗移栽成活率为90%以上.     

中国兰的组织培养

就中国兰组织培养中的形态建成 ,培养基和激素等因素与外植体诱导、种子非共生萌发、原球茎和根状茎的增殖及分化的关系 ,以及中国兰组织培养中存在的问题作了介绍。     

向日葵组织培养研究进展

向日葵(Helianthus annuus)是世界主要的油料作物之一.近几年来有关向日葵的研究很多,其中向日葵组织培养研究越来越受到重视.本文从向日葵外植体培养、植株再生影响因素、组培过程中存在的问题及解决方法等方面进行了综述.     

黄芩愈伤组织诱导和细胞培养研究进展

黄芩是传统中药,近年来,由于其抗癌、抗氧化、抗HIV和清除自由基等功效而格外受到关注.本文综述了黄芩愈伤组织培养中的外植体选择、诱导条件及培养条件选择,以及悬浮细胞培养中次生代谢产物的调控等新进展,以期为大规模培养黄芩细胞生产次生代谢产物提供理论参考和技术借鉴.     

非洲菊组织培养(简报)

以非洲菊嫩叶切段作外植体进行组织培养,与以花托作外植体相比,明显缩短愈伤组织诱导期.该方法尤其适合于种子繁殖(盆栽)的品种.     

枣树组织培养研究进展

综述了枣树的器官培养、愈伤组织培养、花药培养、胚与胚乳培养、原生质体培养以及影响枣树组织培养的其他因素。营养器官培养报道的最多,其他外植体的培养相对较少,研究尚处于起步阶段。还针对枣树组织培养存在问题提出了今后的研究方向。     

变色秋海棠的繁殖栽培

利用播种、扦插和组织培养 3种方式繁殖变色秋海棠均获得成功。播种繁殖的发芽率约 60 % ;在 3种不同的叶插繁殖中 ,以锥形叶插成活率最高 ;组织培养以叶片为外植体、MS+BA1 +NAA0 .1固体发芽培养基较好 ,从外植体直接分化出芽原基和新芽 ,属于器官型再生方式。利用密闭容器栽培法或类似此种方法栽培变色秋海棠 ,取得较好效果。     

‘灿烂’蓝莓组培与快繁技术

以蓝莓良种‘灿烂’半木质化茎段为材料,通过对增殖培养基以及生根培养基的筛选,建立蓝莓高效组培快繁技术体系。结果表明,最佳的增殖培养基为1/2MS＋2.0 mg·L^-1 ZT＋0.01 mg·L^-1 NAA＋30 g·L^-1白糖＋6.5 g·L^-1琼脂,芽诱导率为87%,增殖系数达3.6;ZT 1.0～2.0 mg·L^-1对不定芽诱导效果明显,随着ZT浓度的提高,愈伤组织产生越明显,影响增殖苗的质量。最佳生根培养基为1/2MS＋1.5 mg·L^-1 IBA＋0.01 mg·L^-1 NAA ＋25 g·L^-1白糖＋7.0 g·L^-1琼脂,pH值5.8,生根率70%,IBA浓度从0.5 mg·L^-1提高到1.5 mg·L^-1,生根越多,根系越壮,但也产生更多愈伤组织影响生根质量。炼苗10 d后移栽至草炭土,成活率78%。     

Propagation by partial tissue culture of Austral Bracken (Pteridium esculentum) for revegetation

Austral Bracken (Pteridium esculentum) is a native fern common in many Australian ecosystems and is needed in large numbers for revegetation projects. The main limiting factor for the propagation of locally sourced material is spore availability. A mass propagation system was developed by combining tissue culture and nursery‐based systems. Spores collected over the summer months from wild populations were germinated in vitro on ½ MS medium containing 0.15% w/v activated charcoal. Gametophytes were rapidly multiplied on the same medium. In vitro sporophyte development was unreliable although sometimes prolific. However, gametophytes transferred to a pine bark potting medium with added coir, on a capillary bed in a fog house, produced sporophytes reliably. Across different seasons and populations, 75–100% of the gametophyte explants developed sporophytes within about 9 weeks. Three hundred propagated ferns planted into two field sites within their provenance origins had a survival of 92 and 95% respectively, 3 or 4 months after planting. This report delivers a ready‐to‐use and reliable protocol for the mass propagation of bracken fern of local origin to the revegetation industry.     

Phenology and wintering capacity of sporophytes and gametophytes of ferns native to northern Japan

Summary To investigate life history adaptations to cold climates, the leaf development, sporulation period, growing stage of gametophytes, and the frost and drought resistance of sporophytes and gametophytes of 67 fern species native to Kokkaido were studied. Most ferns common in Hokkaido are summer-green with leaves developing during late May to June and decaying during October. Most of the ferns in Hokkaido sporulate during August to early September. Spores dispersed from June to September germinate before winter begins, forming vegetative prothallia. Gametophytes mature only in the following summer. Thus in Hokkaido the gametophytes as well as perennial sporophytes are exposed to severe winter conditions. In order to correlate the life cycles of temperate ferns with winter cold stress, frost resistance of gametophytes, rhizomes, and leaves of sporophytes were determined. Maximal frost resistance of rhizomes reflects the stress conditions of their habitats: rhizomes of forest understory ferns are damaged at-5°to -17.5°C, epiphytic ferns and ferns of habitats exposed to severe frost sustained temperatures of -20° to-40°C. The leaves of winter-green and evergreen ferns resist frost ranging from -25° to -40°C. The leaves of summer-green ferns are killed by late frost below -5°C. With some exceptions, gametophytes of ferns growing on the forest floor resist frost to -40°C and are much hardier than sporophytes. These results suggest the possible restrictive effects of cold climate on the life span of leaves as well as on the sporulation period. If winter cold is one of the decisive factors for seasonality expression and habitat distribution of ferns, the sensitive generation must be the sporophyte rather than the gametophyte. The hardier gametophyte is therefore able to colonize habitats in which the sporophyte is excluded by frost if mechanisms of vegetative propagation are evolved.Contribution No. 2451 from The Institute of Low Temperature Science     

Anomalous branching patterns in some fossilFilicales: Implications in the evolution of the megaphyll and the lateral branch,habit and growth pattern

From middle Carboniferous coal balls leaves and stems of structurally preserved Filicalean ferns (Psalixochlaenaceae, Botryopteridaceae) are described that deviate from the normal pattern of branching. From this, interpretations are made as to the origin and relationship of axillary branching and dichotomy in this line of ferns. Their megaphylls apparently have orginated from a three-dimensional dichotomous branching system. This is inferred from the widespread occurrence of short, leafy epiphyllous buds on their fronds and is related to their habit where the leaf often plays the dominant role and effects vegetative propagation.     

大蒜分生组织培养脱病毒和快速繁殖技术

采用茎尖分生组织培养技术,获得了大蒜（Ａｌｌｉｕｍ ｓａｔｉｖｕｍ Ｌ．）的无病毒试管苗。通过基本培养基和激素配比试验,筛选出最佳的培养基组成,进行脱病毒苗的快速繁殖。结果表明：诱导愈伤组织的最适培养基为：ＭＳ＋ＢＡ０．２ｍｇ／Ｌ＋ＮＡＡ０．５ｍｇ／Ｌ,月生长率达１２．７０倍;诱导丛生芽的最适培养基为：Ｂ５＋ＢＡ０．５ｍｇ／Ｌ＋ＩＡＡ０．２ｍｇ／Ｌ,丛生芽繁殖系数高达２５．５倍,技术上达到了快速繁     

Symptoms of Fern Distortion Syndrome Resulting from Inoculation with Opportunistic Endophytic Fluorescent Pseudomonas spp.

Background

Fern Distortion Syndrome (FDS) is a serious disease of Leatherleaf fern (Rumohra adiantiformis). The main symptom of FDS is distortion of fronds, making them unmarketable. Additional symptoms include stunting, irregular sporulation, decreased rhizome diameter, and internal discoloration of rhizomes. We previously reported an association of symptoms with increased endophytic rhizome populations of fluorescent pseudomonads (FPs). The aim of the current study was to determine if FPs from ferns in Costa Rica with typical FDS symptoms would recreate symptoms of FDS.

Methodology and Findings

Greenhouse tests were conducted over a 29-month period. Micro-propagated ferns derived from tissue culture were first grown one year to produce rhizomes. Then, using an 8×9 randomized complete block experimental design, 8 replicate rhizomes were inoculated by dipping into 9 different treatments before planting. Treatments included water without bacteria (control), and four different groups of FPs, each at a two concentrations. The four groups of FPs included one group from healthy ferns without symptoms (another control treatment), two groups isolated from inside rhizomes of symptomatic ferns, and one group isolated from inside roots of symptomatic ferns. Symptoms were assessed 12 and 17 months later, and populations of FPs inside newly formed rhizomes were determined after 17 months. Results showed that inoculation with mixtures of FPs from ferns with FDS symptoms, but not from healthy ferns, recreated the primary symptom of frond deformities and also the secondary symptoms of irregular sporulation, decreased rhizome diameter, and internal discoloration of rhizomes.

Conclusions

These results suggest a model of causation of FDS in which symptoms result from latent infections by multiple species of opportunistic endophytic bacteria containing virulence genes that are expressed when populations inside the plant reach a minimum level.     

观赏蕨类引种栽培及其物候期的观察

对中华水韭、福建观音座莲、华南紫萁、西南凤尾蕨、剑叶铁角蕨、翅轴蹄盖蕨、东方荚果蕨、同形鳞毛蕨、圆顶耳蕨等30种蕨类进行了引种栽培和物候期观察.结果表明:(1)引种成活率高;(2)管理粗放;(3)多数种类四季常青,形态优美,具有较高的观赏价值,能够在怀化市安全越冬,值得在亚热带地区开发利用;(4)华南紫萁、小黑桫椤、光蹄盖蕨、长江蹄盖蕨、翅轴蹄盖蕨、三相蕨、同形鳞毛蕨、圆顶耳蕨、镰羽贯众等9种蕨类在展叶时,同时长出孢子囊;(5)多数蕨类的孢子囊在长出后1个月左右发育成熟;(6)在叶开始萌动时引种栽培的蕨类,当年的营养叶萌发期和展叶期都推迟半个月左右,但孢子囊群的出现期和成熟期不受影响;(7)不同蕨类的孢子囊群形成期具有差异性,同种蕨类的孢子囊群形成期具有相对稳定性,因此,可以作为鉴别物种的依据之一。建议在编写、修订《中国植物志》和地方植物志时增加蕨类孢子囊群形成期的描述。     

Biological and nutritional aspects involved in fern multiplication

Gametophytes of several species of ferns were mechanically triturated and the resulting homogenates cultured in vitro for propagation purposes. Differences in the time period from spore culture to sporophyte development were perceivable between species. For those species with a fast life cycle and high sporophyte production such as Woodwardia virginica and Dryopteris affinis sp. affinis, homogenization of gametophytes can be considered to be excellent method for propagation, yielding hundreds of sporophytes in a short period of time. Sporophyte formation was inhibited in O. regalis by the succesive application of homogenization to gametophytes regenerated by this technique. The effect of the culture medium composition on fern production was also studied in O. regalis and P. ensiformis gametophytes. In these species, sporophyte formation increased when the gametophytes were cultured in a medium containing water+0.7% agar. Addition of sucrose inhibited gametophyte development and induced their necrosis. The 1/2 dilution of Murashige and Skoog basal medium, without sucrose, favoured leaf expansion in P. ensiformis sporophytes. This revised version was published online in June 2006 with corrections to the Cover Date.     

湖南壶瓶山国家级保护区药用蕨类植物的调查研究

通过野外调查、标本采集与鉴定和查阅文献,对壶瓶山国家级保护区药用蕨类植物多样性进行统计,并对其资源分布,药用部分、功能分类及民族文化等方面进行了探讨。结果如下：（1）壶瓶山国家级保护区有药用蕨类165种（不包括变种和变型）,隶属于33科57属,分别占武陵山地区和全国药用蕨类总科属种的80.48%7,1.25%,77.10%和67.34%,47.90%3,6.91%;含种数较多的科属有水龙骨科Polypodiaceae,鳞毛蕨科Dryopteridaceae,凤尾蕨科Pteridaceae,卷柏属Selaginella,凤尾蕨属Pteris,鳞盖蕨属Microlepia等。（2）该区药用蕨类植物原始和进化的类群共存,区系以热带成分为主,具有热带亲缘性,体现了壶瓶山药用蕨类植物起源古老,但物种分化强烈的特点。（3）该区的药用蕨类在四个植被垂直带谱都有分布,其中在低于1 000 m的常绿阔叶林带分布最多。（4）该区药用蕨类根据药用部位分为四类,全草类最多,其次是根茎类;按药用功能将分为五类,主要为清热类和解毒类。（5）讨论了壶瓶山国家级保护区独特的地理位置和优厚的自然环境是该区药用蕨类植物多样性高的重要原因,并提出了可持续开发利用与保护的建议。     

Experimental Studies of Apospory in Ferns

Apospory in Pteridium aquilinum was found to have several novelfeatures, in particular the rapidity of its occurrence. Outgrowthsfrom detached laminae were detected in as little as 3 days.There was no evidence that failure of co-ordination betweencells, senescence of the sporophytic tissue or unfavourableculture conditions effected the initiation of outgrowths asearlier workers have suggested. The response of Thelypterispalustris was notably slower and more in line with earlier reportsof apospory. Marsilea vestita appeared to be incapable of anyform of aposporous behaviour. The only feature common to all occurrences of apospory in leavesof wild-type ferns is the severance of the lamina from the mainbody of the plant. It is proposed that this may deprive thecells of regulatory substances which maintain the activationof sporophytic genes. The results are also considered in relationto the phase changes in homosporous and heterosporous plantsand the behaviour of fern protoplasts in culture. apospory, ferns, Pteridium aquilinum, Marsilea vestita, Thelypteris palustris