Pairing and recombination at meiosis of Brassica rapa (AA) × Brassica napus (AACC) hybrids

Interspecific crosses contribute significantly to plant evolution enabling gene exchanges between species. The efficiency of interspecific crosses depends on the similarity between the implicated genomes as high levels of genome similarity are required to ensure appropriate chromosome pairing and genetic recombination. Brassica napus (AACC) is an allopolyploid, resulting from natural hybridization between Brassica rapa (AA) and Brassica oleracea (CC), both being diploid species derived from a common ancestor. To study the relationships between genomes of these Brassica species, we have determined simultaneously the pairing and recombination pattern of A and C chromosomes during meiosis of AAC triploid hybrids, which result from the interspecific cross between natural B. napus and B. rapa. Different AAC triploid hybrids and their progenies have been analysed using cytogenetic, BAC-FISH, and molecular techniques. In 71% of the pollen mother cells, homologous A chromosomes paired regularly, and usually one chromosome of each pair was transmitted to the progeny. C chromosomes remained mainly univalent, but were involved in homoeologous pairing in 21.5% of the cells, and 13% of the transmitted C chromosomes were either recombined or broken. The rate of transmission of C chromosomes depended on the identity of the particular chromosome and on the way the hybrid was crossed, as the male or as the female parent, to B. napus or to B. rapa. Gene transfers in triploid hybrids are favoured between A genomes of B. rapa and B. napus, but also occur between A and C genomes though at lower rates.     

Beneficial endophytic microorganisms of Brassica – A review

Brassica species display enormous diversity and subsequently provide the widest assortment of products used by man from a single plant genus. Many species are important for agriculture, horticulture, in bioremediation, as medicines, soil conditioners, composting crops, and in the production of edible and industrial oils such as liquid fuels and lubricants. Many wild Brassica relatives possess a number of useful agronomic traits, including beneficial microbial endophytes that could be incorporated into breeding programs. Endophytes of Brassica, and/or their metabolites, have been demonstrated to improve and promote plant growth; increase yield; reduce disease symptoms caused by plant pathogens; reduce herbivory from insect pests; remove contaminants from soil; improve plant performance under extreme conditions of temperature and water availability; solubilise phosphate and contribute assimilable nitrogen to their hosts. Brassica napus (oilseed rape) and Brassica oleracea var. botrytis (broccoli and cauliflower) are the most economically important species of Brassica worldwide. These commercial crops are attacked by a wide range of pathogens and insect pests that are responsible for millions of dollars in lost revenue, with current control options offering little mitigation. No alternative control products are available for the Brassica industry, although it has been well documented in the literature that the use of endophytic microorganisms can offer beneficial traits to their host plants, including pest and disease resistance. The aim of this review is to describe the literature concerning beneficial microbial endophytes and their prospects to enhance or provide additional traits to their Brassica host species.     

Just how complex is the Brassica S-receptor complex?

Of the plant self-incompatibility (SI) systems investigated to date, that possessed by members of the Brassicaceae is currently the best understood. Whilst the recent demonstrations of interactions between the male determinant (S-locus cysteine rich protein, SCR) and the female determinant (S-locus receptor kinase, SRK) indicate the minimal requirement for SI in Brassica, no consensus exists as to the nature of these molecules in vivo and the potential involvement of accessory molecules in establishing the active S-receptor complex. Variation between S haplotypes appears to be present in the molecular composition of the receptor complex, the regulation of downstream signalling and the requirement for accessory molecules. This review discusses what constitutes an active receptor complex and highlights potential differences between haplotypes. The role of accessory molecules, in particular SLG (S-locus glycoprotein) and low molecular weight pollen coat proteins (PCPs), in pollination are discussed, as is the link between SI and unilateral incompatibility (UI).     

青菜(Brassica chinensis)花药培养的初步研究

青菜是上海地区主要叶菜类蔬菜,但存在种性退化、病害严重、产量不稳的问题。为了培育抗病、高产的优良品种,我们于1976—1977年对青菜的花药培养进行了初步研究。材料和方法1976年用(二月慢×三月慢)F_1和白叶三月慢、马桥常菜作试验材料;1977年用9个 F_1和两     

油菜(Brassica napus L)收获指数的变异

本文首次采用田间试验的方法对国内油菜收获指数的变异进行了研究．结果表明不同油菜品种的收获指数存在变异,其变异规律大致是：１）中熟品种的收获指数高于早熟品种及晚熟品种;２）中长角果的品种较长角品种具有更高的收获指数;３）高产的新品种具有较高的收获指数;４）油菜粒壳比,角杆比及千粒重与收获指数呈正相关,杆重与收获指数呈负相关．     

油菜（Brassica napusL）的收获系数的变异

本文首次采用田间试验的方法对国同油菜收获指数的变异进行了研究。结果表明不同油菜品种的收获指数存在变异,其变异规律大致是：１）中熟品种的收获指数高于早熟品种及晚熟品种;２）中长角果的品种较长角品种具有更高的收获指数;３）高产的新品种具有较高的收获指数;４）油菜粒壳比,角杆比及千粒重与收获指数呈正相关,杆重与收获指数呈负相关。     

Genetic load and transgenic mitigating genes in transgenic Brassica rapa (field mustard) × Brassica napus (oilseed rape) hybrid populations

Background

Cell transplantation is likely to become an important therapeutic tool for the treatment of various traumatic and ischemic injuries to the central nervous system (CNS). However, in many pre-clinical cell therapy studies, reporter gene-assisted imaging of cellular implants in the CNS and potential reporter gene and/or cell-based immunogenicity, still remain challenging research topics.

Results

In this study, we performed cell implantation experiments in the CNS of immunocompetent mice using autologous (syngeneic) luciferase-expressing bone marrow-derived stromal cells (BMSC-Luc) cultured from ROSA26-L-S-L-Luciferase transgenic mice, and BMSC-Luc genetically modified using a lentivirus encoding the enhanced green fluorescence protein (eGFP) and the puromycin resistance gene (Pac) (BMSC-Luc/eGFP/Pac). Both reporter gene-modified BMSC populations displayed high engraftment capacity in the CNS of immunocompetent mice, despite potential immunogenicity of introduced reporter proteins, as demonstrated by real-time bioluminescence imaging (BLI) and histological analysis at different time-points post-implantation. In contrast, both BMSC-Luc and BMSC-Luc/eGFP/Pac did not survive upon intramuscular cell implantation, as demonstrated by real-time BLI at different time-points post-implantation. In addition, ELISPOT analysis demonstrated the induction of IFN-γ-producing CD8+ T-cells upon intramuscular cell implantation, but not upon intracerebral cell implantation, indicating that BMSC-Luc and BMSC-Luc/eGFP/Pac are immune-tolerated in the CNS. However, in our experimental transplantation model, results also indicated that reporter gene-specific immune-reactive T-cell responses were not the main contributors to the immunological rejection of BMSC-Luc or BMSC-Luc/eGFP/Pac upon intramuscular cell implantation.

Conclusion

We here demonstrate that reporter gene-modified BMSC derived from ROSA26-L-S-L-Luciferase transgenic mice are immune-tolerated upon implantation in the CNS of syngeneic immunocompetent mice, providing a research model for studying survival and localisation of autologous BMSC implants in the CNS by real-time BLI and/or histological analysis in the absence of immunosuppressive therapy.     

油菜(Brassica napus L.)叶片植株的再生

油菜是一种重要的油料作物,有关它的组织培养方面的研究已有许多报道。但由叶片诱导成再生植株的不多,且系由单倍体或无菌苗而来,时间较长,取材受限,不便应用。因此,有必要研究自然条件下的二倍体叶片植株的再生及其条件,特别是良种,以加速其无性繁殖,并探索细胞、组织分化及其形态发生的规律和机理。故1980年来我们作     

油菜(Brassica napus L.)花茎组织培养中的器官形成

用甘蓝型油菜的主花茎切段,比较了不同激素和组合对器官形成的调节作用,花茎不同部位及花茎不同组织分化能力的差异。结果表明单加BA(2毫克/升)对芽的形成有明显促进作用。在加BA 的培养基中再加入NAA(0.05或0.5毫克/升)均显著降低形成芽的百分率及芽的数目。B_5培养基的效果明显优于MS 配方。花茎不同部位切段的比较试验表明,在花茎中芽形成能力在茎顶部5厘米中最强,往下渐减。将花茎表皮和皮层与髓部组织分开进行培养,则芽形成能力显著下降,但对根和愈伤组织的形成影响较小。油菜花茎所具有的强烈再生能力可能为维持和加速繁殖优良单株,或不育系等育种材料提供技术。     

油菜（Brassica napus）油酸性状的遗传规律研究

以高油酸油菜品系04-863(油酸含量80.5%)与低油酸油菜品系湘油15号自交系04-1020(油酸含量62.7%)为材料,通过杂交获f1、F2代和回交一代,研究了油菜油酸性状的遗传规律.结果表明:1)油菜油酸性状属数量性状.呈连续变异.很难明确分组;2)杂种一代表现为两亲本的中间类型,但受母本的影响较大,当以高油酸品系04-863为母本时,F1代的油酸含量(X=73.41)分布倾向高油酸亲本,相反,若以低油酸湘油15号自交系04-1020为母本,F1代油酸含量(X=67.75)分布则倾向低油酸亲本;3)对环境条件变化比较敏感,亲代(P1、P2)或F1代都存在着由于环境条件差异而引起的表型差异.F2代广泛分离既有基因型分离的差异,又有环境引起的表型差异.度量这种差异分别以方差和标准差来表示离开平均数的变异程度,在各世代中以F2的方差和标准差最大,分别为V=52.81,S=7.28;4)各代变异的次数分布大体上符合正态分布,即极端类型个体少,中间类型多;5)支配油菜油酸性状的基因数为一对主基因符合孟德尔的3:1分离规律,其回交比例为1:1,油酸性状的遗传力为69%～71%,是由于遗传差异引起的,另31%～29%则是由于环境差异引起的.说明数量性状与质量性状的区分是相对的.     

Molybdenum sequestration in Brassica species. A role for anthocyanins?

To elucidate plant mechanisms involved in molybdenum (Mo) sequestration and tolerance, Brassica spp. seedlings were supplied with molybdate, and the effects on plant physiology, morphology, and biochemistry were analyzed. When supplied with (colorless) molybdate Indian mustard (Brassica juncea) seedlings accumulated water-soluble blue crystals in their peripheral cell layers. Energy dispersive x-ray analysis showed that Mo accumulated predominantly in the vacuoles of the epidermal cells. Therefore, the blue crystals are likely to be a Mo compound. The x-ray absorption spectrum of the plant-accumulated Mo was different than that for molybdate, indicating complexation with a plant molecule. Because the blue compound was water soluble and showed a pH-dependent color change, possible involvement of anthocyanins was investigated. An anthocyanin-less mutant of Brassica rapa ("fast plants") was compared with varieties containing normal or high anthocyanin levels. The anthocyanin-less mutant did not show accumulation of a blue compound when supplied with molybdate. In the anthocyanin-containing varieties, the blue compound colocalized with anthocyanins in the peripheral cell layers. Mo accumulation by the three B. rapa varieties was positively correlated with anthocyanin content. Addition of molybdate to purified B. rapa anthocyanin resulted in an in vitro color change from pink to blue. Therefore, Mo appears to be sequestered in vacuoles of the peripheral cell layers of Brassica spp. as a blue compound, probably a Mo-anthocyanin complex.     

Poly(β-hydroxybutyrate) production in oilseed leukoplasts of Brassica napus

Polyhydroxyalkanoates (PHAs) comprise a class of biodegradable polymers which offer an environmentally sustainable alternative to petroleum-based plastics. Production of PHAs in plants is attractive since current fermentation technology is prohibitively expensive. The PHA homopolymer poly(β-hydroxybutyrate) (PHB) has previously been produced in leaves of Arabidopsis thaliana (Nawrath et al., 1994, Proc Natl Acad Sci USA 91: 12760–12764). However, Brassica napus oilseed may provide a better system for PHB production because acetyl-CoA, the substrate required in the first step of PHB biosynthesis, is prevalent during fatty acid biosynthesis. Three enzymatic activities are needed to synthesize PHB: a β-ketothiolase, an acetoacetyl-CoA reductase and a PHB synthase. Genes from the bacterium Ralstonia eutropha encoding these enzymes were independently engineered behind the seed-specific Lesquerella fendleri oleate 12-hydroxylase promoter in a modular fashion. The gene cassettes were sequentially transferred into a single, multi-gene vector which was used to transform B. napus. Poly(β-hydroxybutyrate) accumulated in leukoplasts to levels as high as 7.7% fresh seed weight of mature seeds. Electron-microscopy analyses indicated that leukoplasts from these plants were distorted, yet intact, and appeared to expand in response to polymer accumulation. Received: 26 May 1999 / Accepted: 16 June 1999     

Assigning Brassica microsatellite markers to the nine C-genome chromosomes using Brassica rapa var. trilocularis–B. oleracea var. alboglabra monosomic alien addition lines

Brassica rapa var. trilocularis-B. oleracea var. alboglabra monosomic alien addition lines (MAALs) were used to assign simple sequence repeat (SSR) markers to the nine C-genome chromosomes. A total of 64 SSR markers specific to single C-chromosomes were identified. The number of specific markers for each chromosome varied from two (C3) to ten (C4, C7 and C9), where the designation of the chromosomes was according to Cheng et al. (Genome 38:313-319, 1995). Seventeen additional SSRs, which were duplicated on 2-5 C-chromosomes, were also identified. Using the SSR markers assigned to the previously developed eight MAALs and recently obtained aneuploid plants, a new Brassica rapa-B. oleracea var. alboglabra MAAL carrying the alien chromosome C7 was identified and developed. The application of reported genetically mapped SSR markers on the nine MAALs contributed to the determination of the correspondence between numerical C-genome cytological (Cheng et al. in Genome 38:313-319, 1995) and linkage group designations. This correspondence facilitates the integration of C-genome genetic information that has been generated based on the two designation systems and accordingly increases our knowledge about each chromosome. The present study is a significant contribution to genetic linkage analysis of SSR markers and important agronomic traits in B. oleracea and to the potential use of the MAALs in plant breeding.     

青花菜(Brassica oleracea)品种"上海2号"花枝

诱导和增殖培养基:(1)MS 6-BA1.0 mg·L-1(单位下同) NAA0.1;(2)MS 6-BA1.0 NAA0.2;(3)MS 6-BA 1.0 NAA 0.5;(4)MS 6-BA0.5; NAA0.1     

芸苔属（Brassica）植物中microRNAs的生物信息学预测与分析

MicroRNA (miRNA) 是一类调控基因转录后表达的非编码的小分子RNA．它在生物的发育、细胞增殖、凋亡以及胁迫响应等生物过程中发挥着重要的调控作用．目前,分离和鉴定miRNA的方法主要包括实验方法（遗传筛选、直接克隆）和生物信息学方法．MiRNA存在表达丰度低,表达组织特异性,以及受特殊诱导等问题,用传统的实验法常难发现和鉴定miRNA．通过生物信息学方法在已有的各种基因库中寻找未知miRNA,大大提高了人们发现miRNA及其靶基因的效率．芸苔属（Brassica）的成员包括油菜、芜青、甘蓝等,是世界各国主要油料和食用作物．目前,油菜的miRNA的分离和鉴定工作已有文献报道,而其它的尚属空白．本文将拟南芥、水稻等植物已知的miRNA分别与芜青、甘蓝、野芥菜、黑芥菜、埃塞俄比亚芥的GSS和EST数据库进行比对搜索,采用一系列标准进行筛选,最后分别在芜青和甘蓝中预测到67个和95个miRNA．再把这些预测得到的miRNA分别与芜青和甘蓝的mRNA数据库进行比对搜索,分别找到120个和111个靶基因,除去未知功能及功能不详的,各有62个和48个靶基因．分析结果表明,上述大多数靶基因编码的产物为转录因子及重要代谢酶类,涉及植物的生长发育调控,信号转导及胁迫响应等方面．     

Access and Identification of Intergeneric Hybrid Between Brassica juncea×Crambe abyssinica

An intergeneric hybrid between Brassica juncea (L.) Coss. x Crambe abyssinica Hochst. ex R. E. Fries was obtained for the first time through routine method combined with embryo rescue. The inductivity of hybrids was 2.1%. In many respects the Fl hybrid resembled either its maternal or paternal parent br both. Consequenfiy, retarded growth, wrinkled leaves and rudimentary anthers in Fl plant were observed. Fl pollen varied in size and 10% ~ 15% were of the paternal pollen type,i.e. ,4-colpate. Fl pollen viability was low and 12% ~ 25% of pollens was viable, with a mean of 18.5%. The chromosome number of Fl somatic cell was 63 which was intermediate between the two parents. Most of pollen mother cells had 63 chromosomes. Analysis of isozymes further confirmed that the F1 hybrid combined the paternal characteristic bands.     

Functional characterization of β-ketoacyl-CoA synthase genes from Brassica napus L.

Seed-specifically expressed -ketoacyl-CoA synthase genes of Brassica napus (Bn-FAE1.1 genes) were cloned from two cultivars, namely Askari, a high-erucic-acid type, and Drakkar, a low-erucic-acid type. The genes from the two cultivars were found to be nearly identical. They encode proteins of 507 amino acids, the sequences of which differ only at position 282. The Bn-FAE1.1 gene of Askari, unlike that of Drakkar, was functionally expressed in yeast cells suggesting that the single amino acid exchange effects the low erucic acid phenotype at the E1 gene locus. In yeast cells the -ketoacyl-CoA synthase of Askari elongated not only oleoyl but also palmitoleoyl groups as well as saturated acyl groups in such a way that monounsaturated acyl groups of 22 carbons and saturated ones of 26 carbons were formed as main products. A reporter gene fused to the promoter region of the Bn-FAE1.1 gene from Askari showed seed-specific expression in transgenic rapeseed plants. Over-expression of the coding region of the Askari gene in developing seeds of transgenic Drakkar plants resulted in a significant increase in the levels of eicosenoic acid and erucic acid esterified in the seed oil. On the other hand, in transgenic high-erucic-acid rapeseed plants the increase in erucic acid level was at most 60% although the chimeric Bn-FAE1.1 gene was co-expressed with an erucoyl-CoA-specific lysophosphatidate acyltransferase gene enabling trierucoyl glycerol to accumulate in the seed oil.     

青菜(Brassica chinesis L.)对碘动态吸收的同位素示踪

碘是合成甲状腺激素不可缺少的元素,食物中严重缺碘可以导致人体中甲状腺功能的失调。利用同位素示踪技术研究了青菜在水培条件下对碘（^125Ⅰ）的吸收和富集特征,并在此基础上讨论了植物对碘吸收的生物学机制。研究结果表明,青菜根都能迅速吸收^125Ⅰ并向地上部分输送,青菜各部位^125Ⅰ的比活度大小顺序是根〉茎〉叶;青菜茎对碘的吸收量白天大于夜晚,青菜叶对碘的吸收夜晚大于白天;青菜植株不同部位叶片碘含量存在差异,上部嫩叶中^125Ⅰ高于下部叶片,说明青菜中^125Ⅰ的富集特征是植物对碘主动吸收和被动吸收共同作用的结果。青菜培养时间经过120h后,^125Ⅰ的消耗途径主要是作物的吸收和自然挥发,^125Ⅰ的放射性衰变丢失的量相对较少。研究为培育含碘作物提供了科学依据。     

Molecular Cloning and Characterization of Enolase from Oilseed Rape （Brassica napus）

An enolase-encoding cDNA clone in oilseed rape (Brassica napus L.) was isolated. This gene (accession number: AY307449) had a total length of 1 624 bp with an open reading frame of 1 335 bp, and encoded a predicted polypeptide of 444 amino acids with a molecular weight of 47.38 kD. The deduced amino acid sequence shared identity with a number of enolases ranging from Bacillus subtilis to human beings and had much higher identity with other plant enolases than with enolases from Bacillus, yeast and human beings. Comparison of its primary structure with those of other enolases revealed the presence of an insertion of five amino acids in enolase of B. napus. Southern blotting analysis of genomic DNA indicated that enolase was likely to be a low-copy gene in the oilseed rape genome. Expression of the cloned enolase gene increased under salt stress, but decreased in response to low temperature. Our studies suggested that the cloned gene was a new member of plant enolase gene family, which contributed to the energy supply in stress-treated tissues.