江西铅山红芽芋胚性愈伤组织的玻璃化法超低温保存及植株再生

以江西铅山红芽芋胚性愈伤组织为材料,研究各种因素对其玻璃化法超低温保存的影响。结果表明:江西铅山红芽芋胚性愈伤组织玻璃化法超低温保存较佳的预培养条件为0.3mol·L-1蔗糖预培养3d,较佳的60%PVS2装载时间为20min,较佳的100%PVS2脱水条件为25℃脱水30min,较佳的化冻温度为40℃,较佳的洗涤液蔗糖浓度为1.2mol·L-1,较佳的冻后培养条件为暗培养7d再转到光周期中培养。红芽芋胚性愈伤组织包埋玻璃化超低温保存后的平均成活率约为70%。红芽芋胚性愈伤组织冻后再生苗没有发生形态学、生理学和细胞学的变异。     

江西铅山红芽芋胚性愈伤组织的包埋玻璃化超低温保存

为长期安全保存江西铅山红芽芋种质资源,本文以江西铅山红芽芋的胚性愈伤组织为对象,研究了包埋玻璃化冻存过程中各因素对细胞活力和愈伤组织成活率的影响,优化建立了江西铅山红芽芋胚性愈伤组织包埋玻璃化超低温保存体系。将约0.2 g胚性愈伤组织块包埋成海藻酸钙凝胶珠后,在25℃下转入MS+2 mg/L TDZ+1 mg/L NAA+0.75 mol/L蔗糖的培养基中于14 h/d光周期下预培养1 d;预培养后的胚性愈伤组织块用2 mol/L甘油和0.4 mol/L蔗糖的混合物在25℃下装载40 min;采用PVS2在25℃下脱水30 min,更换PVS2后直接投入液氮保存1 d;再将胚性愈伤组织块置于37℃恒温水浴中化冻3 min,然后用MS+2 mg/L TDZ+1 mg/L NAA+1.2 mol/L蔗糖的液体培养基洗涤3次,每次10 min;洗涤后的胚性愈伤组块转入MS+2 mg/L TDZ+1mg/L NAA固体培养基上先暗培养7 d再转到14 h/d光周期中培养。7 d后胚性愈伤组织块开始恢复生长,并且在30 d内分化出胚状体;将胚状体再次转入MS+2 mg/L TDZ+1 mg/L NAA固体培养基上,60 d后形成完整的植株。红芽芋胚性愈伤组织包埋玻璃化超低温保存后的平均成活率约为60%,并且红芽芋胚性愈伤组织冻后再生苗没有发生形态性状和染色体数目的变异,此结果为长期安全保存江西铅山红芽芋种质资源奠定了良好的基础。     

江西铅山红芽芋胚性愈伤组织小滴玻璃化法超低温保存技术的研究

探索江西铅山红芽芋胚性愈伤组织的小滴玻璃化法超低温保存,为其种质资源的超低温保存提供技术基础和理论依据。植物组织培养和单因子试验的方法。江西铅山红芽芋胚性愈伤组织小滴玻璃化法超低温保存的较佳程序为:约0.2 g胚性愈伤组织在25℃下转入MS+2 mg·L-1TDZ+1 mg·L-1NAA+0.5 mmol·L-1蔗糖的培养基中,于14 h·d-1光周期下预培养3 d后用MS+2 mmol·L-1甘油+0.4 mmol·L-1蔗糖在25℃下装载20min,然后用PVS2在0℃下脱水40 min,吸取5滴PVS2到铝箔条上,将脱水后的红芽芋胚性愈伤组织块转到铝箔条上的PVS2液滴里。附有胚性愈伤组织块的铝箔条在液氮里蘸一下,然后迅速将其转入2 mL装满液氮的冷冻管中,再投入液氮保存1 d。从液氮中取出冷冻管中的铝箔条,浸入用40℃温水预热过的洗涤液(MS+2 mg·L-1TDZ+1 mg·L-1NAA+1.2 mmol·L-1蔗糖)中,使胚性愈伤组织块从铝箔条上脱落下来,然后再将胚性愈伤组织块转入新鲜的洗涤液中洗涤3次,每次10 min。洗涤后转入MS+2 mg·L-1TDZ+1 mg·L-1NAA固体培养基上,先暗培养7 d再转到14 h·d-1光周期中培养。30 d后将分化出的胚状体再次转入MS+2 mg·L-1TDZ+1 mg·L-1NAA固体培养基上可再生完整植株。红芽芋胚性愈伤组织包埋玻璃化超低温保存后的平均成活率约为80%。红芽芋胚性愈伤组织冻后再生苗没有发生形态学、生理学和细胞学的变异。红芽芋胚性愈伤组织小滴玻璃化法超低温保存可以保证其遗传资源的稳定性,为江西铅山红芽芋种质资源的离体保存提供了一条新的途径。     

大苞鞘石斛原球茎玻璃化超低温保存技术的研究

本文研究建立了大苞鞘石斛(Dendrobium wardianum Warner)原球茎玻璃化法超低温保存的技术体系。结果发现,预处理和玻璃化溶液(plant vitrification solution 2,PVS2)装载脱水是影响大苞鞘石斛原球茎相对存活率的两个关键步骤,高渗与低温-高渗两种预处理方法测定的相对存活率具有显著性差异;玻璃化溶液的种类以及脱水时间对冻后存活率具有重要的影响。基于此,建立了大苞鞘石斛原球茎的超低温保存体系,即:以继代培养60 d的大苞鞘石斛原球茎为材料,1/2MS+0.8 mol/L蔗糖的培养基上4℃低温预处理6 d后,转至1/2 MS+2 mol/L甘油+0.4 mol/L蔗糖的装载液中室温下装载40 min,在0℃下装载PVS2脱水40 min,然后转入装有新鲜PVS2冷冻管中并迅速投入液氮。在液氮保存1 h后放在40℃水浴中快速解冻1 min,利用含1.2 mol/L蔗糖的1/2MS培养液洗涤3次,每次间隔10 min;待恢复培养30 d后统计存活率,可使大苞鞘石斛原球茎超低温保存后存活率达到20.0%。     

花烛胚性悬浮细胞玻璃化超低温保存条件研究

为建立适宜的花烛(Anthurium andraeanum Lind. )胚性悬浮细胞玻璃化超低温保存技术,采用单因素实验方法对影响玻璃化超低温保存后细胞相对存活率的主要因素进行了研究.结果表明,经玻璃化超低温保存后花烛悬浮细胞的相对存活率与悬浮细胞的继代培养时间、渗透调节剂的种类和浓度及预培养时间、装载液种类和预处理时间、PVS2脱水时间以及超低温保存后的化冻温度均有一定的关系.继代培养3和5 d,细胞的相对存活率较高(约20%);分别以0.3、0.5、0.7 mol·L-1山梨醇和60、80、100、120 g·L-1蔗糖为渗透调节剂预培养0～4 d,以0.5 mol·L-1山梨醇预培养2 d的效果最好,细胞的相对存活率为26.2%;用体积分数25%PVS2预处理15 min,细胞的相对存活率最高(29.0%);分别用体积分数100%PVS2脱水0、5、10、15、20、25和30 min,其中脱水10 min的悬浮细胞相对存活率最高(32.1%);分别在10 ℃、20 ℃、30 ℃、40 ℃、50 ℃和60 ℃水浴条件下进行化冻处理,其中用40 ℃水浴化冻的悬浮细胞相对存活率最高(32.1%).花烛胚性悬浮细胞玻璃化超低温保存和化冻的适宜流程为:将继代培养3～5 d的胚性悬浮细胞团(直径2 mm)在含0.5 mol·L-1山梨醇的1/2MS液体培养基中预培养2 d后,于4 ℃条件下处理24 h,然后先用体积分数25%PVS2室温预处理15 min,再用体积分数100%PVS2 在0 ℃条件下脱水10 min,最后迅速投入液氮中冷冻保存;将经过冷冻保存的细胞置于40 ℃水浴中化冻3 min,用含1.2 mol·L-1蔗糖的1/2MS液体培养基洗涤3次(每次10 min),之后即可进行恢复培养.     

江西铅山红芽芋胚性愈伤组织的包埋干燥法超低温保存

对江西铅山红芽芋（Colocasia esculenta var．cormosus cv．Hongyayu）胚性愈伤组织包埋干燥法超低温保存进行了初步的研究。结果表明：江西铅山红芽芋胚性愈伤组织包埋干燥法超低温保存较佳的条件为：0．75mol·L-1蔗糖预培养3d;脱水方式为空气干燥7h或硅胶干燥11h;化冻温度为37℃（2min）;冻后培养条件为暗培养7d再转到光周期中培养。此方法超低温保存后的平均成活率约为45％。超低温保存时间以及是否去除包裹的褐藻酸钙对其成活率无显著性影响。形态学和细胞学检测表明红芽芋胚性愈伤组织冻后再生苗与母本材料相比没有发生变异。     

野生稻愈伤组织的超低温保存和冻后再生植株的形成

对不同基因组的野生稻愈伤组织进行了超低温保存的研究,主要结果如下：①野生稻愈伤组织经过预培养→预处理→冰冻降温→液氮保存→快速解冻的超低温保存,冻后细胞存活率最高可达８７．９％。②１０％ＤＭＳＯ＋８％葡萄糖为最佳冰冻保护剂。降温程序为０℃→－１０℃,１５ｍｉｎ→－４０℃,６０ｍｉｎ→液氮（ＬＮ）。③普通野生稻、宽叶野生稻、疣粒野生稻获得了冻后再生植株。④疣粒野生稻解冻后形成旺盛胚性愈伤组织,并通过体细胞胚胎发生途径再生出大量植株。     

野葛试管苗茎尖玻璃化法超低温保存及植株再生

对野葛茎尖玻璃化法超低温保存技术的研究结果表明,H号野葛茎尖较佳的保存技术体系是:继代生长30 d的野葛无菌苗置于4℃冰箱炼苗5 d;在无菌条件下切取含1~2个叶原基(1.5~2.5 mm)的茎尖,转至含5% DMSO+5%蔗糖的MS培养基内,置于4℃冰箱预培养1 d;用60%、100% PVS₂(30%甘油+15%乙二醇+15% DMSO+13.7%蔗糖)分别在0℃下过渡和脱水各30 min,随即迅速投入液氮;保存24 h后,在40℃水浴中快速化冻90 s,用含41.1%蔗糖的MS培养液洗涤2次,每次停留10 min;转至再生培养基(MS+6-BA 2 mg/L+NAA 1 mg/L)暗培养7 d,然后光下培养,再生率可达60%以上。再生苗与常温苗形态指标差异不显著。     

长鞭红景天悬浮培养细胞的玻璃化法超低温保存研究

对长鞭红景天悬浮培养细胞的玻璃化法超低温保存进行了初步研究.结果表明,预培养、预处理、脱水处理及冻后处理对长鞭红景天悬浮培养细胞存活率均有重要影响,方差分析结果均显示差异显著.长鞭红景天悬浮培养细胞过程中最佳培养条件是:在含5%二甲基亚砜(DMSO)的MS培养基上预培养1 h,室温下80%PVS2预处理40 min,然后用100%PVS2于0℃处理50 min,投入液氮(LN)保存1 h后在40℃水浴中迅速化冻,再用1.2 mol/L蔗糖培养液洗涤3次,每次10 min,洗涤后的悬浮培养细胞用氯化三苯四氮唑(TTC)法检测,其存活率可达72.70%.     

枇杷叶片愈伤组织的诱导与保存

在不同成熟度的枇杷叶片中,最适宜诱导愈伤组织的是中度成熟叶;最适合的愈伤组织诱导培养基是MS+6-BA 1.0mg/L+2,4-D 0.5mg/L,诱导出的愈伤组织为黄绿色、颗粒状、硬度适中;适合愈伤组织继代保存的培养基为MS+6-BA 0.5~1.0mg/L+2,4-D 0.1~0.2mg/L。     

Foreign exploration for Scirtothrips perseae Nakahara (Thysanoptera: Thripidae) and associated natural enemies on avocado (Persea americana Miller)

Scirtothrips perseae Nakahara was discovered attacking avocados in California, USA, in 1996. Host plant surveys in California indicated that S. perseae has a highly restricted host range with larvae being found only on avocados, while adults were collected from 11 different plant species. As part of a management program for this pest, a “classical” biological control program was initiated and foreign exploration was conducted to delineate the home range of S. perseae, to survey for associated natural enemies and inventory other species of phytophagous thrips on avocados grown in Mexico, Guatemala, Costa Rica, the Dominican Republic, Trinidad, and Brazil. Foreign exploration efforts indicate that S. perseae occurs on avocados grown at high altitudes (>1500 m) from Uruapan in Mexico south to areas around Guatemala City in Guatemala. In Costa Rica, S. perseae is replaced by an undescribed congener as the dominant phytophagous thrips on avocados grown at high altitudes (>1300 m). No species of Scirtothrips were found on avocados in the Dominican Republic, Trinidad, or Brazil. In total, 2136 phytophagous thrips were collected and identified, representing over 47 identified species from at least 19 genera. The significance of these species records is discussed. Of collected material 4% were potential thrips biological control agents. Natural enemies were dominated by six genera of predatory thrips (Aeolothrips, Aleurodothrips, Franklinothrips, Leptothrips, Scolothrips, and Karnyothrips). One genus each of parasitoid (Ceranisus) and predatory mite (Balaustium) were found. Based on the results of our sampling techniques, prospects for the importation of thrips natural enemies for use in a “classical” biological control program in California against S. perseae are not promising.     

Molecular phylogeny of Phebalium (Rutaceae: Boronieae) and related genera based on the nrDNA regions ITS 1+2

Parsimony analyses of the internal transcribed spacer regions of nuclear ribosomal DNA (ITS 1 & ITS 2) for 38 taxa sampled from the Phebalium group (Rutaceae: Boronieae) and two outgroups confirm that, with the exception of Phebalium sensu stricto and Rhadinothamnus, six of the currently recognised genera within the group are monophyletic. The data indicate that Phebaliums. str. is paraphyletic with respect to Microcybe, and Rhadinothamnus is paraphyletic with respect to Chorilaena. Rhadinothamnus and Chorilaena together are the sister group to Nematolepis. Drummondita, included as an outgroup taxon, clustered within the ingroup as sister to Muiriantha and related to Asterolasia.The phylogeny suggests that the evolution of major clades within a number of these genera (e.g. Phebalium) relates to vicariance events between eastern and south-western Australia. Leionema is an eastern genus, with the most basal taxon being the morphologically distinct Leionema ellipticum from northern Queensland. Leionema also includes one species from New Zealand, but this species (as with some others) proved difficult to sequence and its phylogenetic position remains unknown. Taxonomic changes at the generic level are recommended.The authors wish to thank Paul G.Wilson, PERTH, for advice and discussion, and Paul Forster, BRI, for collecting and providing material of Leionema ellipticum. The project was supported by a Melbourne University Postgraduate Award (to BM), the Australian Biological Resources Study (ABRS), Australian Systematic Botany Society and Wolf Den (Australia) Investments.     

New species and new combinations in Peruvian Orchidaceae

seventeen new species and combinations are proposed in the generaChondrorhyncha, Cischweinfia, Cochlioda, Eloyella, Encyclia, Kefersteinia, Koellensteinia, Macroclinium, Rodriguezia, Solenidiopsis, andStenia. All new species are illustrated. A key is provided for 2-flowered species ofMacroclinium, PeruvianSigmatostalix, and PeruvianStenia. Solenidium (Solenidiopsis) peruvianum Schltr. is lectotypified.     

Entwicklungsgeschichte und Ultrastruktur von Pollenkitt und Exine bei nahe verwandten entomophilen und anemophilen Angiospermensippen derAlismataceae,Liliaceae, Juncaceae,Cyperaceae, Poaceae undAraceae

Some closely related members of the monocotyledonous familiesAlismataceae, Liliaceae, Juncaceae, Cyperaceae, Poaceae andAraceae with variable modes of pollination (insect- and wind-pollination) were studied in relation to the ultrastructure of pollenkitt and exine (amount, consistency and distribution of pollenkitt on the surface of pollen grains). The character syndromes of pollen cementing in entomophilous, anemophilous and intermediate (ambophilous or amphiphilous) monocotyledons are the same in principal as in dicotyledons. Comparing present with former results one can summarize: 1) The pollenkitt is always produced in the same manner by the anther tapetum in all angiosperm sub-classes. 2) The variable stickiness of entomophilous and anemophilous pollen always depends on the particular distribution and consistency of the pollenkitt, but not its amount on the pollen surface. 3) The mostly dry and powdery pollen of anemophilous plants always contains a variable amount of inactive pollenkitt in its exine cavities. 4) A step-by step change of the pollen cementing syndrome can be observed from entomophily towards anemophily. 5) From the omnipresence of pollenkitt in all wind-pollinated angiosperms studied one can conclude that the ancestors of anemophilous angiosperms probably have been zoophilous (i.e. entomophilous) throughout.

     

安徽牯牛降的丝孢菌Ⅰ

本文报导作者采自安徽枯牛降自然保护区的18种丝孢菌,分属于5个属,其中有3个新种:牛皮冻生尾孢(Cercospora paederiicola),山鸡椒假尾饱(Pseudocercospora litseae-cubebae),鸡血藤生假尾孢(P. millettiicola)和2个中国新纪录。文中对新种进行了描述及绘图,新记录种作了简要说明。研究的标本保存在中国科学院微生物研究所真菌标本室(HMAS)。     

Changes of dung beetle communities from rainforests towards agroforestry systems and annual cultures in Sulawesi (Indonesia)

Little is known about how tropical land-use systems contribute to the conservation of functionally important insect groups, including dung beetles. In a study at the margin of Lore Lindu National Park (a biodiversity hotspot in Central Sulawesi, Indonesia) dung-beetle communities were sampled in natural forest, young secondary forest, agroforestry systems (cacao plantations with shade trees) and annual cultures (maize fields), each with four replicates (n = 16 sites). At each site we used 10 pitfall traps, baited with cattle dung, along a 100 m transect for six 3-day periods. The number of trapped specimens and species richness at the natural forest sites was higher than in all land-use systems, which did not significantly differ. Each land-use system contained, on average, 75% of the species richness of the natural forest, thereby indicating their importance for conservation. However, a two-dimensional scaling plot based on NESS indices (m = 6) indicated distinct dung beetle communities for both forest types, while agroforestry systems and annual cultures exhibited a pronounced overlap. Mean body size of dung beetles was not significantly influenced by land-use intensity. Five of the six most abundant dung beetle species were recorded in all habitats, whereas the abundance of five other species was significantly related to habitat type. Mean local abundance and number of occupied sites were closely correlated, further indicating little habitat specialisation. The low dung beetle diversity (total of 18 recorded species) may be due to the absence of larger mammals in Sulawesi during historical times, even though Sulawesi is the largest island of Wallacea. In conclusion, the dung beetle fauna of the lower montane forest zone in Central Sulawesi appears to be relatively robust to man-made habitat changes and the majority of species did not exhibit strong habitat preferences.     

Taxonomic status of the species of the green algal genusNeochloris

Komárek has recently reviewed the various species assigned to the green algal genusNeochloris Starr (Chlorococcales, Chlorococcaceae) and removed those with uninucleate vegetative cells to a new genus,Ettlia. Watanabe & Floyd, unaware ofKomárek's work, also reviewed the species ofNeochloris and distributed them among three genera—Neochloris, Chlorococcopsis gen. nov., andParietochloris gen. nov.—on the basis of details of the covering of the zoospore and the arrangement of the basal bodies of the flagellar apparatus. This paper reconciles these two treatments and makes additional recommendations at the ranks of genus, family, order, and class.     

Phylogenetic investigations of Sordariaceae based on multiple gene sequences and morphology

The family Sordariaceae incorporates a number of fungi that are excellent model organisms for various biological, biochemical, ecological, genetic and evolutionary studies. To determine the evolutionary relationships within this group and their respective phylogenetic placements, multiple-gene sequences (partial nuclear 28S ribosomal DNA, nuclear ITS ribosomal DNA and partial nuclear β-tubulin) were analysed using maximum parsimony and Bayesian analyses. Analyses of different gene datasets were performed individually and then combined to generate phylogenies. We report that Sordariaceae, with the exclusion Apodus and Diplogelasinospora, is a monophyletic group. Apodus and Diplogelasinospora are related to Lasiosphaeriaceae. Multiple gene analyses suggest that the spore sheath is not a phylogenetically significant character to segregate Asordaria from Sordaria. Smooth-spored Sordaria species (including so-called Asordaria species) constitute a natural group. Asordaria is therefore congeneric with Sordaria. Anixiella species nested among Gelasinospora species, providing further evidence that non-ostiolate ascomata have evolved from ostiolate ascomata on several independent occasions. This study agrees with previous studies that show heterothallic Neurospora species to be monophyletic, but that homothallic ones may have a multiple origins. Although Gelasinospora and Neurospora are closely related and not resolved as monophyletic groups, there is insufficient evidence to place currently accepted Gelasinospora and Neurospora species into the same genus.     

Die GattungKarschia — Bindeglied zwischen bitunicaten Ascomyceten und lecanoralen Flechtenpilzen?

The genusKarschia, in the earlier sense, including saprophytes and parasites on lichens, has been thought to be a non-lichenized parallel genus of the lichen genusBuellia. Modern workers included it on the one hand inBuellia, on the other hand combined it with bitunicate ascomycetes. It is now proved thatKarschia is heterogeneous and contains but superficially similar members both of the genusBuellia of theLecanorales and of typical or masked bitunicateAscomycetes. Therefore, it can not be regarded as a link betweenLecanorales andDothideales. The type species ofKarschia belongs to theDothideales.