芥蓝和结球甘蓝染色体组型及C-带带型的研究

本文用改进的染色体标本制片技术,研究了芥蓝和结球甘蓝的染色体组型和 C-带带型。两种植物的二倍体均由4对中着丝粒、5对亚中着丝粒染色体组成,其中一对为随体染色体。芥蓝和结球甘蓝具有统一的染色体组型公式:2n=18=8m+10sm(2SAT),但两者的某些染色体在编号顺序上有差异。在结球甘蓝中观察,到4种不同形态的随体。用 BSG C-带方法得到 C-带带型,带型公式,芥蓝为2n=18=CITS 型=10C+2CI_++4CT~++2CS;结球甘蓝为2n=18=CITS 型=8c+2CI_++6CT~++2CS。某些带纹具多态性和杂合性。本文从染色体水平上讨论了芥蓝与甘蓝的亲缘关系。     

埃塞俄比亚芥与芥蓝杂交获得异源三倍体及其细胞学研究

以埃塞俄比亚芥(2n=4x=BBCC=34)和芥蓝(2n=2x=CC=18)为材料,通过相互杂交获得了异源三倍体(2n=3x=BCC=26)。该异源三倍体生长势较强;叶色等介于双亲之间;株型、花型和花大小偏向于埃塞俄比亚芥;花色与芥蓝的相同,为白花。减数分裂观察表明:在终变期,一般形成9个二价体和8个单价体(9Ⅱ+8Ⅰ),且B、C两组染色体表现出一定程度的分群现象;中期Ⅰ,CC基因组的9个二价体排列在赤道板上,而B组的8个单价体游离在赤道板周围;后期Ⅰ分到两极的染色体以13/13和12/14占多数,偶见落后的染色体。该BCC异源三倍体的获得为创建CC+B染色体的异附加系和研究B、C基因组间的亲缘关系奠定了基础。     

Chromosome complement, C-banding and Ag-NOR in Gymnammodytes cicerelus (Ammodytidae, Perciformes)

The karyotype of a sandlance species, Gymnammodytes cicerelus , comprises: seven meta-centric, seven submetacentric and nine subtelocentric-acrocentric pairs (2 n =46, FN=74). The C-bands appear in paracentromeric and telomeric areas of most chromosomes and the NOR regions, in two pairs of larger chromosomes. All these characteristics indicate that a large number of rearrangements seem to have been involved in the karyotype evolution of this species.     

农杆菌介导的芥蓝遗传转化体系的建立

采用正交旋转设计方法对影响芥蓝遗传转化体系的因素进行了优化研究,结果表明:影响芥蓝KanR苗率的最主要因素是Kan浓度,而预培养时间和共培养时间是芥蓝遗传转化的主要影响因素。最利于芥蓝遗传转化的操作程序为:将芥蓝无菌苗下胚轴在预培养基上预培养2天后,在LBA4404菌液中感染8min,置于共培养培养基上培养2天,随后把外植体转入含Kan的选择分化培养基上诱导不定芽,28天转瓶一次,当抗性幼苗长至2～3cm时,齐愈伤组织处切下幼苗在生根培养基上诱导不定根,25天左右后等不定根长成即可开瓶炼苗,继而移栽至营养土中,正常管理至开花结果;经PCR、Southern印迹检测,证明CYP86MF基因已经整合至转基因植株染色体中。     

两种鼠耳蝠的核型、G带和C带研究

使用肺、心脏等组织进行培养,用空气干燥法制作染色体标本,以胰酶法制作G带,BSG法制作c带,分析了贵州2种鼠耳蝠的核型、G带和c带.水鼠耳蝠Myotis daubentoni和小鼠耳蝠Myotis dividii的染色体数均为2n=44,拥有3对大型和1对中型中着丝粒染色体,染色体臂数(FN)=52;这2种鼠耳蝠的G带...     

大白菜生物技术研究进展

综述了国内外在大白菜(Brassica rapa ssp.pekinensis)单倍体培养、基因工程和分子标记方面的最新研究成果.重点讨论了大白菜小孢子培养的影响因素、取材要点、培养条件、植株再生和倍性鉴定以及大白菜的转基因载体选择、受体系统建立、遗传转化与转基因植株鉴定等.对小孢子培养及转基因研究的原理、方法和关键技术进行了总结.展望了大白菜生物技术研究的应用前景.     

Construction of random sheared fosmid library from Chinese cabbage and its use for Brassica rapa genome sequencing project

As a part of the Multinational Genome Sequencing Project of Brassica rapa, linkage group R9 and R3 were sequenced using a bacterial artificial chromosome (BAC) by BAC strategy. The current physical contigs are expected to cover approximately 90% euchromatins of both chromosomes. As the project progresses, BAC selection for sequence extension becomes more limited because BAC libraries are restriction enzyme-specific. To support the project, a random sheared fosmid library was constructed. The library consists of 97536 clones with average insert size of approximately 40 kb corresponding to seven genome equivalents, assuming a Chinese cabbage genome size of 550 Mb. The library was screened with primers designed at the end of sequences of nine points of scaffold gaps where BAC clones cannot be selected to extend the physical contigs. The selected positive clones were end-sequenced to check the overlap between the fosmid clones and the adjacent BAC clones.Nine fosmid clones were selected and fully sequenced. The sequences revealed two completed gap filling and seven sequence extensions, which can be used for further selection of BAC clones confirming that the fosmid library will facilitate the sequence completion of B. rapa.     

硅对受土壤中镉污染的白菜生长和抗胁迫能力的影响

在土培盆栽条件下,硅可提高白菜的地上部鲜重和茎鲜重,但根鲜重下降,叶鲜重略有下降。硅可抑制白菜吸收镉,在0.3、0.6、1.2mg·kg~(-1)镉水平下,施硅可显著降低白菜地上部的镉含量,并在一定程度上提高叶中过氧化物酶(POD)活性,但降低超氧化物歧化酶(SOD)活性。0.5、1.0 g·kg~(-1)硅可提高白菜的叶绿素含量和过氧化氢酶(CAT)活性,降低叶细胞膜透性,从而提高其对镉胁迫的耐受力。较高的硅浓度对植物有一定的毒性,叶绿素含量和CAT活性都下降,细胞膜透性也增加。     

白菜OguCMS相关MYB家族新基因BcMYBogu的克隆与特征分析

为研究CMS核质互作的分子机理, 将甘蓝型油菜(Brassica napus L.)和白菜(B. campestris L. ssp. chinensis Makino)杂交并连续回交6代获得白菜OguCMS, 在与保持系花药细胞学比较的基础上, 运用cDNA-AFLP筛选得到白菜OguCMS早、中期花蕾提早表达的MYB-like差异片断, 利用RACE克隆得到该片断的cDNA全长, 命名为BcMYBogu(GenBank 登录号: EF127861), 对其氨基酸序列和表达特征进行研究。结果表明, 白菜OguCMS绒毡层在四分体后增生, 高度液泡化, 导致小孢子花粉外壁异常, 细胞质同外壁分离并降解; 花药变白; BcMYBogu具有典型的MYB DNA结合域—W残基和SH[AL]QKY[RF]基序; 系统进化分析显示BcMYBogu与AtMYB26, AtMYB32和AtMYB4等聚类在同一分枝; RT-PCR分析表明BcMYBogu在莲座叶、花茎和花蕾中均有表达, 但在OguCMS花蕾中表达量显著上升。由此推测BcMYBogu是一个新的与白菜OguCMS相关的MYB家族新成员。     

Construction of random sheared fosmid library from Chinese cabbage and its use for Brassica rapa genome sequencing project

As a part of the Multinational Genome Sequencing Project of Brassica rapa,linkage group R9 and R3 were sequenced using a bacterial artificial chromosome(BAC) by BAC strategy.The current physical contigs are expected to cover approximately 90%euchromatins of both chromosomes.As the project progresses,BAC selection for sequence extension becomes more limited because BAC libraries are restriction enzyme-specific.To support the project,a random sheared fosmid library was constructed.The library consists of 9...     

Selection for resistance to clubroot (Plasmodiophora brassicae) in marrowstem kale (Brassica oleracea var. acephala L.)

Ninety-six cultivars of Brassica oleracea were screened for clubroot resistance in a seedling test using two populations of Plasmodiophora brassicae. The most resistant cultivars were kales. Sixteen resistant marrowstem kale cultivars of diverse geographical origin were used to start a selection programme for clubroot resistance. Four generations of selection, involving single plants, half-sib and full-sib families, reduced a disease index averaged over six clubroot populations from 41.2 to 12.5. This was lower than the most resistant cultivar in the original population, cv. Mixti 28.8, and as good as a German landrace of cabbage noted for its resistance, Bohmerwaldkohl 10.5. In comparison, the mean of five kale controls, cvs Bittern, Canson, Condor, Kestrel and Merlin, was 61.1 and the value for the most susceptible control, cabbage cv. Septa, was 89.3. In the final assessment, there were no clubroot population x B. oleracea genotype interactions and in the initial assessment of cultivars there were only small interactions which could be removed by an angular transformation of the data. It was concluded that a high level of non-differential resistance had been achieved and that it may prove durable. It was also concluded from a small field trial that this level of resistance would prevent serious yield losses in practice.     

Selection for high and low thiocyanate ion content in kale (Brassica oleracea var. acephala)

In 1978 genetical variation for thiocyanate ion (SCN-) was found in a kale population being selected for improved digestible organic-matter yield. Four generations of half-sib family selection for high and low SCN^- content in young leaves resulted in subpopulations with almost a twofold difference in their means, no overlap in their distributions, no observable reduction in their genetical variation, and only small differences in other traits. The total observed divergence in SCN^-content was 82% of that expected for a random mating population in linkage equilibrium in the absence of genotype x environment interactions. Average heritability on a single plot basis was low, namely 30%, but was increased to 46% with two replicates. This maximised the predicted response to selection for a fixed number of families selected and a fixed total number of plots.
Bulks of the high and low subpopulations were assessed in a trial in 1988. Averaged over September and December harvests of young and mature leaves, SCN- contents were 104.5 and 58.5 mg/100 g DM, respectively. Of eight controls, cv. Merlin had the lowest content, 63.5, and cv. Proteor had the highest content, 89.6. Cultivar x harvest date, cultivar x leaf age and cultivar x harvest date X leaf age interactions were removed by a logarithmic transformation of the data. Stem contents were very low and variable with an overall mean of 10.9 in September and > 5 in December, and no significant differences between cultivars or populations.
The subpopulations will be valuable for research into possible effects of SCN content on the disease and pest resistance of kale and on the performance of animals fed kale.     

大白菜细胞质雄性不育保持系3411-7--RAPD特异扩增片段的克隆测序

利用RAPD技术从大白菜细胞质雄性不育保持系3411-7的DNA中得到一个特异扩增片段MOPB04600。回收该特异扩增片段并将其克隆到pGEM-TEasy载体上进行序列测定。结果表明该片段全长600bp,其碱基组成为A+T=72.33%。通过与GenBank+EMBL+DDBJ+PDB中的455,972个序列进行同源性比较,同源性均小于30%,表明该片段为一新发现的序列。并对该特异片段的来源及其可能的作用进行探讨。     

High frequency plant regeneration from stem explants of Brassica alboglabra Bailey in vitro

Culture conditions for shoot regeneration and proliferation, and rooting of Brassica alboglabra Bailey were optimized by a judicious selection of explants and manipulation of hormonal combinations in the culture medium. Both half and whole stem explants were more regenerative than cotyledons and hypocotyls. The highest shoot regeneration frequency (100%) accompanied by high number of shoots was obtained using half stem explants grown on Murashige & Skoog [14] medium supplemented with 2 mgl^-1 BA in combination with 1 mgl^-1 NAA, or 4 mgl^-1 2iP with 0.5 mgl^-1 NAA. For shoot proliferation, 4 mgl^-1 kinetin was most effective. The presence of auxin reduced shoot proliferation significantly. Maximum rooting (100%) of shoot cuttings was obtained either in the absence or in the presence of 0.5 mgl^-1 NAA, or IBA or IAA ranging from 0.1 to 8 mgl^-1.     

醌还原酶法进行芥蓝抗癌活性的快速检测

研究了不同提取方法,不同细胞裂解剂和醌还原酶(QR)反应液的培育时间对QR法分析的影响。结果表明,芥蓝组织用磷酸缓冲液(5mmol/LK2HPO4-KH2PO4,1mmol/LEDTA,pH7.6)提取效果良好,与常用的三元试剂提取法效果相当。这种方法简单而有效,优于其他有机溶剂提取法。0.4%乙基苯基聚乙二醇(NP40)可以替代细胞裂解剂——0.8%毛地黄皂苷用于QR分析,且效果良好,QR反应中反应液的培育时间以5￣10min为宜。     

The karyotype of Adenomera diptyx (Boettger 1885) (Anura, Leptodactylidae) from northeastern Argentina

In this work we analyzed the karyotype of five populations of Adenomera diptyx from Argentina after conventional staining, Ag-NOR and C-banding. All specimens presented 2n = 26 and FN = 34. The karyotype was formed by three submetacentric, one metacentric and nine telocentric pairs. Silver staining revealed that the NOR was located on a secondary constriction in pair 7. C- banding evidenced constitutive heterochromatin at the pericentromeric region of all chromosomes. The karyotype of A. diptyx was similar to that of A. hylaedactyla (2n = 26, FN = 34) and different from that of A. andreae (2n = 26, FN = 40) in the fundamental number and secondary constriction position. It also differed from the karyotypes of A. marmorata (2n = 24, FN = 34 and 36) and of A. aff. bokermanni (2n = 23, FN = 34) in diploid number. Until a comprehensive cytogenetic analysis of all the species of the genus is performed, their chromosome evolution will remain poorly understood.     

Glucosinolate responses of oilseed rape, mustard and kale to mechanical wounding and infestation by cabbage stem flea beetle (Psylliodes chrysocephala)

Mechanical wounding of the petioles of six laboratory-grown rapeseed ( Brassica napus ) cultivars induced physiological changes in the plant, markedly affecting the levels of individual glucosinolates. Greatest increases were observed for the indole glucosinolates, glucobrassicin and neoglucobrassicin. Such changes were usually associated with large decreases in the levels of aliphatic glucosinolates. The total glucosinolate content of the wounded plant was thus a reflection of these two opposing trends and wounding produced a greater relative indole glucosinolate content in this total figure. Thus increasing wounding was associated with an increase in indole glucosinolates and a decrease in aliphatic compounds.
Infestation of field- and laboratory-grown rapeseed with cabbage stem flea beetle ( Psylliodes chrysocephala ) produced similar effects, which were observed in various parts of the plant. Differences in response between field- and laboratory-grown infested plants are attributed to the different physiological ages of the harvested material.
Laboratory-grown kale and mustards also showed wound-induced glucosinolate changes. The kale, cv. Fribor, produced elevated levels of both indoles and aliphatics after wounding. Total glucosinolate content in the mustards, which, unlike rape and kale, normally contain only traces of indole glucosinolates in the unstressed state, was increased following wounding. This was, however, not associated with elevated levels of indole glucosinolates, but with accumulation of aliphatic ( Brassica nigra, B. juncea ) and aromatic ( Sinapis alba ) glucosinolates. The significance of these findings is discussed.