虎眼万年青离体快繁体系及无糖生根培养

以虎眼万年青的叶片为外植体,对不同激素浓度的愈伤组织和不定芽的诱导情况以及再生芽的无糖培养情况进行了研究.结果表明:适于愈伤组织诱导及不定芽分化的培养基为MS+BA1.0mg/L+NAA0.1mg/L,适宜的继代培养基为MS+BA0.5mg/L+NAA0.1mg/L;用无糖培养体系进行生根培养时,基质中NAA浓度为0.1mg/L时,再生芽的生根率达94%,种苗质量优于常规组培.     

Evaluation of Ornithogalum genebank accessions for some characteristics of importance for breeding cut flowers or pot plants

The South African Ornithogalum L. species are divided into three subgenera; Aspasia, Urophyllon and Osmyne. The species of horticultural importance, such as O. thyrsoides, are found in the subgenus Aspasia, and all have pure coloured flowers ranging from white to yellow to deep orange. Accessions numbering 299 were collected and grown under comparative conditions. Characteristics of use in discriminating superior parents for hybridization were measured, such as: leaf position and colour; stem length; shape, size and density of the inflorescence; size, shape and colour of the flower; the presence of visual virus symptoms and the reaction of leaf cuttings to tissue culture propagation. The aim was to quantify the variation according to the characteristics deemed of importance for selecting genotypes for cut or pot flower breeding. Variation for each of the traits measured was observed in each of the species. The evaluation of the Ornithogalum improved the process of selecting accessions as parents in hybridization by providing an indication of the variation present in the natural populations. Thus specific accessions could be selected to combine either at interspecific or intraspecific level to breed for distinct types.     

预处理对巴西盾柱木种子吸涨、活力测试及其加速老化的影响

对巴西盾柱木(Peltophorum dubium)种子进行了4种处理的吸胀实验,即将种子分别置于10℃和27℃下0.2% Captan溶液中和-1.0MPa的PEG6000溶液中,每个处理有四个重复,40粒种子,置于培养皿中浸满实验溶液的双层过滤纸上,覆以透气良好的薄膜.种子在实验前均用98%的浓硫酸处理15分钟以打破机械休眠.吸胀曲线显示,种子在吸胀过程中最终重量的增加值为70%-150%,以后进入一个停滞期.在5种处理下(对照组,PEG6000溶液中10℃和27℃下预处理,Captan溶液中10℃和27℃下预处理),对种子萌发所受到的影响进行了实验.每一个处理分别进行3个次级处理,即在实验前,将种子用蒸馏水分别浸泡12、24和36小时.萌发率最高的是经过蒸馏水浸泡12小时对照组种子和PEG 27℃组的种子,萌发率达到100%;萌发率最低的是蒸馏水中浸泡36小时的PEG 27℃组的种子,萌发率为52%;经过蒸馏水浸泡24小时的PEG 10℃组种子的平均萌发时间为1.08天.蒸馏水中浸泡12小时的PEG 27℃组种子平均萌发时间为2.42天,其它处理的萌发时间值介于两者之间.对预处理和未经预处理种子通过加速老化实验,对其活力和生存力进行了测试.经72小时加速老化,种子萌发率低或没有萌发力.对照组种子在培养皿(27℃)和室温条件下蛭石中的萌发率都比处理组高,表现了对老化更强的抗性.     

虎眼万年青提取物体外清除自由基活性的研究

[目的]研究虎眼万年青不同体积分数乙醇提取物体外抗氧化作用。[方法]采用邻二氮菲-Fe2＋氧化法、邻苯三酚自氧化法（MTT）和DPPH法分别研究不同体积分数乙醇对虎眼万年青提取物抗氧化活性的影响。[结果]结果表明虎眼万年青乙醇提取物对.OH、O 2-.和DPPH.均有清除作用,且清除能力与其含虎眼万年青总皂苷含量成正比。不同体积分数的虎眼万年青乙醇提取物体外抗氧化能力的大小为70%乙醇提物〉80%乙醇提物〉60%乙醇提取物〉40%乙醇提取物〉20%乙醇提取物。[结论]虎眼万年青皂苷类成分与其抗氧化性有关。     

Characterization of a novel potyvirus tentatively named <Emphasis Type="Italic">Ornithogalum</Emphasis> virus 3, successfully isolated from <Emphasis Type="Italic">O. thyrsoides</Emphasis> co-infected with two other potyviruses by single-aphid inoculation

A potyvirus tentatively named Ornithogalum virus 3 (OV-3) was successfully isolated by single-aphid transmissions from O. thyrsoides mix-infected with OV-3, Ornithogalum mosaic virus (OrMV) and Ornithogalum stripe mosaic virus (OrSMV). OV-3, a flexuous, rod-shaped particle of ca. 690 nm, was sap and aphid transmissible. The virus had a narrow host range and caused necrotic mosaic on O. thyrsoides under cold conditions. We therefore propose the name Ornithogalum necrotic mosaic virus (OrNMV) for OV-3. A synergistic increase in symptom severity was apparent on O. thyrsoides mix-infected with OrSMV/OrNMV, but not with either OrMV/OrNMV or OrMV/OrSMV. The nucleotide sequence data reported is available in the DDBJ/EMBL/GenBank databases under accession number AB282754.     

不同碳源对虎眼万年青离体生长的影响

［目的］研究不同碳源对虎眼万年青离体生长的影响。［方法］以虎眼万年青子鳞茎为外植体,以MS培养基为基本培养基,附加6BA1.0mg/L、NAA0.2mg/L,在培养基中分别加入不同浓度的蔗糖（15、30、45g/L）、麦芽糖（15、30、45g/L）和葡萄糖（15、30、45g/L）。［结果］蔗糖为虎眼万年青离体生长的最佳碳源。当蔗糖浓度为30g/L时,虎眼万年青离体生长的繁殖系数最高,长势最好。［结论］为从根本上解决虎眼万年青生产过程中优化品种、周年供应、控制成本等问题提供理论依据。     

番红花种球中虎眼万年青花叶病毒的分子鉴定与序列分析

【目的】对一批荷兰进境番红花种球(Crocus sativus)进行病毒鉴定,以防止危险性病毒传入危害。【方法】根据已报道的马铃薯Y病毒科(Potyviridae)通用引物(Sprimer和M4T),采用RT-PCR方法对一批番红花种球样品进行检测,取其中一个RT-PCR产物进行克隆测序,利用Gen Bank上的基本局部比对搜索工具(Basic Local Alignment Search Tool,BLAST)和MEGA 6中的基于对数期望的多重序列比较(Multiple Sequence Comparison by Log-Expectation,MUSCLE)进行序列比对分析,并根据外壳蛋白基因(coat protein,CP)序列利用最大似然法(maximum likelihood method)进行系统发育分析。【结果】RT-PCR检测结果显示该批样品扩增到一条约1.7 kb的预期大小片段,说明该批番红花种球样品中存在Potyviridae科病毒。测序获得长度为1 666bp的序列(命名为2677-NL),含有部分核内含体b基因(nuclear inclusion b,NIb)、完整的CP和3′端非编码区(3′-UTR),其核苷酸序列与虎眼万年青花叶病毒(Ornithogalum mosaic virus,Or MV)的SW3.3分离物具有最高序列一致性(99.6%),与Or MV参考基因组序列(Reference genomic sequences,Ref Seq;NC_019409)的序列一致性为99.1%。该分离物的CP与6个Or MV印度分离物(JQ686722、JQ686720、JN692498、JN692497、JN692496、JF682235)的核苷酸和氨基酸序列一致性分别为71.5%—77.3%和65.7%—79.7%,与其他32个Or MV分离物的核苷酸和氨基酸序列一致性分别为77.9%—99.5%和82.9%—99.2%。基于CP进行的系统发育分析表明,Or MV可分为两个类群,而2677-NL与5个澳大利亚分离物(JQ807995、JQ807996、NC_019409、AF185964、AF185965)相聚成簇,表明其在系统发育关系上与Or MV澳大利亚分离物的亲缘关系最近。【结论】根据国际病毒分类委员会(ICTV)关于Potyviridae科病毒种类的分类标准,2677-NL为Or MV的一个分离物,证实该批番红花种球受到Or MV的侵染。这是世界上首次在番红花中发现Or MV的报道。     

虎眼万年青花粉的形态观察与活力测定

通过光学显微镜与电子扫描显微镜对虎眼万年青的花粉粒进行了观察，发现其花粉粒为两沟型，表面具穿孔，无外萌发器。通过染色方法并结合萌发试验证明其花粉活性低。     

虎眼万年青组织培养中抗生素抑菌作用的研究

[目的]筛选适合虎眼万年青组织培养的最优抗生素。[方法]以虎眼万年青的子鳞茎为外植体,以不加抗生素的处理为对照,向培养基中分别加入不同浓度的四环素、青霉素和链霉素,研究抗生素的抑菌作用。[结果]3种抗生素对细菌污染均有一定抑制效果,在接种初期,抑制作用较为明显,但随着培养时间的延长,抑制作用逐渐降低。从整体看,链霉素的抑制效果较好,但四环素40 mg/L和青霉素20 mg/L的抑制效果更显著。3种抗生素对真菌的抑菌效果是显著的,尤其是青霉素20和40 mg/L。抗生素对外植体生长并无明显的影响。[结论]青霉素20 mg/L无论是在抑制细菌、真菌还是在诱导率方面均优于其他各浓度的几种抗生素,在花卉的快繁方面可以应用。     

Isolation and characterization of a novel potyvirus tentatively named Ornithogalum virus 2

A novel potyvirus, tentatively named Ornithogalum virus 2 (OV-2) because only its nucleotide sequence of the coat protein gene has been revealed, was isolated for the first time from Ornithogalum thyrsoides. OV-2 had a flexuous particle (700–740 nm in length) and was sap and aphid transmissible. The virus had a narrow host range; of 36 test plants in 12 families, only O. thyrsoides and O. dubium were infected. Because the virus caused characteristic stripe mosaic on O. thyrsoides, we propose Ornithogalum stripe mosaic virus (OrSMV), instead of OV-2 for the proper name of the virus. The nucleotide sequence data reported is available in the DDBJ/EMBL/GenBank databases under accession number AB271783.