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.     

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.     

芥菜(Brassica juncea)花粉胚状体与离体种胚脂肪酸的变化

在油菜改良品质育种中,利用单个花粉胚状体筛选低或无芥酸株系,需要了解花粉胚状体在发育过程中芥酸含量的变化及与其他脂肪酸变化的关系。Gurr(1972),Dasgupta等(1973)和黄尚琼(1980)研究了不同种、不同品种种子发育过程中脂肪酸组成的变化,但均未曾对离体花粉胚状体和离体种胚进行研究。本文在于探索离     

Isolation and Preliminary Characterization of Differential Displayed cDNAs in Brassica juncea Exposed to Heavy Metal

Heavy metal contamination of soil has become a major environmental concern worldwide over the past several decades,the exploitation and application of heavy metal hyperaccumulating plants will defend the safety of food and environment on which human life relied,whereas using natural hyperaccumulators is insufficient,due to     

Somatic cell hybridization of ’oxy’ CMS Brassica juncea (AABB) with B. oleracea (CC) for correction of chlorosis and transfer of novel organelle combinations to allotetraploid brassicas

Alloplasmic lines of cultivated Brassica species with B. oxyrrhina cytoplasm are male-sterile and suffer from severe chlorosis. We developed male-sterile lines corrected for chlorosis by fusing protoplasts of CMS B. juncea (AABB) with ’oxy’ cytoplasm and normal B. oleracea (CC). A large number of male-sterile AABBCC somatic hybrids with desirable organelle combinations, i.e. chloroplasts of B. oleracea and mitochondria with recombinant genomes, were recovered. While no recombination was observed in the chloroplast genome, the mitochondrial genome showed extensive recombination that resulted in the appearance of totally novel banding patterns in some of the hybrids. Hybrids with a parental-type mitochondrial genome as well as recombinant patterns close to either of the parental types were also obtained. Using AABBCC somatic hybrids as bridging material, we transferred the desirable organelle combinations to B. juncea (AABB), B. napus (AACC), and B. carinata (BBCC). Many of these lines are now at advanced stages of backcrossing and show stable inheritance of the CMS character and do not suffer from chlorosis. Received: 9 August 1999 / Accepted: 14 September 1999     

Induction, Purification and Characterization of α-1, 3-Glucanase from Brassica juncea L Infected with Albugo candida

Infection of Brassica juncea plants with Albugo candida caused the induction of β-1,3-glucanase in the leaf tissues following inoculation; the induction being more pronounced in the resistant cultivars. The enzyme purified from infected leaves of resistant cultivar RC-781 had a molecular mass of 35.4 kD. The enzyme exhibited its optimum activity at 50°C and at pH 5.5 with K_m value of 0.55 mg laminarin ml^?1. The enzyme activity was slightly stimulated by Cl^? and Mg⁺⁺, while inhibited by Hg⁺⁺ and Mn++ at higher concentrations only     

Sulfur–selenium–molybdenum interactions distinguish selenium hyperaccumulator Stanleya pinnata from non-hyperaccumulator Brassica juncea (Brassicaceae)

Long-term sulfate, selenate and molybdate accumulation and translocation were investigated in two ecotypes of Stanleya pinnata and non-hyperaccumulator Brassica juncea under different levels of applied sulfate and selenate. Morphological differences were observed between the ecotypes of S. pinnata, but few differences in selenium (Se) and sulfur (S) accumulation were measured. Se-to-S ratios were nearly identical between the ecotypes under all treatments. When compared with B. juncea, several unique trends were observed in the hyperaccumulators. While both S. pinnata ecotypes showed no significant effect on Se content of young leaves when the supplied sulfate in the growth medium was increased tenfold (from 0.5 to 5 mM), the Se levels in B. juncea decreased 4- to 12-fold with increased sulfate in the growth medium. Furthermore, S. pinnata’s S levels decreased slightly with high levels of supplied Se, suggesting competitive inhibition of uptake, while B. juncea showed higher S levels with increasing Se, possibly due to up-regulation of sulfate transporters. Both ecotypes of S. pinnata showed much larger Se concentrations in young leaves, while B. juncea showed slightly higher levels of Se in older leaves relative to young. Molybdenum (Mo) levels significantly decreased in S. pinnata with increasing sulfate and selenate in the medium; B. juncea did not show the same trends. These findings support the hypothesis that S. pinnata contains a modified sulfate transporter with a higher specificity for selenate.     

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     

Overcoming prefertilization barriers in the crossDiplotaxis siettiana × Brassica juncea using irradiated mentor pollen

Hybridization between the nearly extinct speciesDiplotaxis siettiana andBrassica juncea is prevented because of strong prefertilization barriers. Use of mentor pollen ofD. siettiana irradiated with 1000 Gy gamma radiation before the incompatible pollination led to fertilization. 5 d after pollination 17% ovules showed entry of pollen tubes, 10 d after pollination 27% ovules showed small globular embryos which grew no further. No embryos were found in control pollinations. Thus , use of irradiated mentor pollen brings about fertilization in this difficult cross and hybrids can be obtained if embryos are rescued.     

Production and cytogenetics of the intergeneric hybrids Brassica juncea×Orychophragmus violaceus and B. carinata×O. violaceus

 Intergeneric hybrids between Brassica juncea (2n=36), B. carinata (2n=34) and Orychophragmus violaceus (2n=24) were produced when B. juncea and B. carinata cultivars were used as female parents. The hybrids between B. juncea and O. violaceus had an intermediate morphology except for petal colour and were partially fertile. The hybrids between B. carinata and O. violaceus had a matroclinous morphology and were nearly fertile. Cytological analysis of the hybrids and their progenies gave the following results. (1) In the hybrids between B. juncea and O. violaceus, the somatic tissues of the roots, leaves and styles were mixoploid (2n=12–42), and cells with 24, 30 or 36 chromosomes were the most frequent. Based on the recorded numbers and behaviour of the mitotic and meiotic chromosomes, complete and partial separation of the parental genomes was proposed to have occurred during mitosis. This resulted in the occurrence of cells with possibly complete and incomplete complements of the parental species and cells with parental complements and some additional chromosomes from the other parent. (2)  Pollen mother cells (PMCs) possibly with both parental chromosome complements, only B. juncea chromosomes or a complete B. juncea complement with additional O. violaceus chromosomes were more competitive in entering meiosis. The majority of fertile gametes were deduced to have been produced by PMCs with a B. juncea complement with or without additional O. violaceus chromosomes. (3) The progeny plants from selfed hybrids between B. juncea and O. violaceus were morphologically either of a B. juncea, hybrid or variable type. Cytologically they were grouped into six types according to the frequencies of cells with various chromosome numbers. All of the plants except 2 which constituted two types, were mixoploids, composed of cells with various chromosome numbers, mainly in a certain serial range. (4) The hybrid plants between B. carinata and O. violaceus were mixoploids with chromosome numbers in the range of 12–34, and cells with 2n=34 were the most frequent. The main categories of PMCs with 17 bivalents at metaphase I and 17 : 17 segregations at anaphase I contributed to the high fertility of the hybrids and the fact that their progeny after selfing were mainly plants with 2n=34. Somatic and meiotic separation of the parental genomes was proposed to have occurred in the hybrids between B. carinata and O. violaceus. (5) Mitotic and meiotic elimination of what could be O. violaceus chromosomes might also have contributed to the observed mitotic and meiotic cell types in the two kinds of hybrids studied. Finally, the possible mechanisms behind these cytological observations and their potential in the production of Brassica aneuploids were discussed. Received: 4 February 1997/Accepted: 29 July 1997     

EDTA and Pb—EDTA accumulation in Brassica juncea grown in Pb—amended soil

Previous studies have shown that EDTA is necessary to solubilize soil Pb and facilitate its transport from the soil to the above ground plant tissues. These studies have also suggested that Pb is accumulated in the plant tissue with transpiration as the driving force. We conducted further studies to evaluate the relationship between EDTA soil treatment, plant transpiration, and plant accumulation of Pb and EDTA. Indian mustard (Brassica juncea) plants were grown in soils containing Pb at three different concentrations (1.5, 3.0 and 4.8 mmol/kg) for 5 weeks before being treated with EDTA concentrations ranging from 0 to 10 mmol/kg. Plant shoots and xylem sap were collected and analyzed for Pb and EDTA content using ICP and HPLC, respectively. Water loss was measured for 7 days following EDTA application. Transpiration was not affected at <5 mmol/kg EDTA but, at 10 mmol/kg EDTA transpiration decreased by 80%, whereas accumulation of Pb and EDTA increased. In the Sassafras soil, Pb and EDTA accumulation in the plant shoots continued to increase as the applied EDTA concentration increased, except at the highest level (10 mmol/kg). In soil amended with 4.8 mmol/kg Pb and 10 mmol/kg EDTA, the concentrations of EDTA and Pb in shoots decreased and visible signs of phytotoxicity were observed. The results presented herein support recent studies in hydroponic systems showing that EDTA and Pb are taken up by the plant and suggest that Pb is translocated in the plant as the Pb-EDTA complex. The results also show that the maximum Pb accumulation by plants occurs by maximizing the concentration of the Pb-EDTA complex based on the EDTA extractable soil Pb. This revised version was published online in July 2006 with corrections to the Cover Date.     

A Moricandia arvensis– based cytoplasmic male sterility and fertility restoration system in Brassica juncea

 A cytoplasmic male-sterility system has been developed in mustard (Brassica juncea) following repeated backcrossings of the somatic hybrid Moricandia arvensis (2n=28, MM)+B. juncea (2n=36, AABB), carrying mitochondria and chloroplasts from M. arvensis, to Brassica juncea. Cytoplasmic male-sterile (CMS) plants are similar to normal B. juncea; however, the leaves exhibit severe chlorosis resulting in delayed flowering. Flowers are normal with slender, non-dehiscent anthers and excellent nectaries. CMS plants show regular meiosis with pollen degeneration occurring during microsporogenesis. Female fertility was normal. Genetic information for fertility restoration was introgressed following the development of a M. arvensis monosomic addition line on CMS B. juncea. The additional chromosome paired allosyndetically with one of the B. juncea bivalents and allowed introgression. The putative restorer plant also exhibited severe chlorosis similar to CMS plants but possessed 89% and 73% pollen and seed fertility, respectively, which subsequently increased to 96% and 87% in the selfed progeny. The progeny of the cross of CMS line with the restorer line MJR-15, segregated into 1 fertile : 1 sterile. The CMS (Moricandia) B. juncea, the restorer (MJR-15), and fertility restored F₁ plants possess similar cytoplasmic organellar genomes as revealed by ‘Southern’ analysis. Received: 17 September 1997 / Accepted: 18 February 1998     

A complex interplay of Gβ and Gγ proteins regulates plant growth and defence traits in the allotetraploid Brassica juncea

Plant Molecular Biology - Gene expression analysis coupled with in-planta studies showed that specific Gβγ combination regulates plant growth and defence traits in the allotetraploid...     

Effects of wild-type and α-tocopherol-enriched transgenic Brassica juncea on the components of xenobiotic metabolism, antioxidant status, and oxidative stress in the liver of mice

Alpha (α)-tocopherol is the most biologically active and preferentially retained form of vitamin E in the human body and is known for its antioxidant and gene regulatory functions. Its increased intake is implicated in protection against diseases that involve an oxidative stress component. We have evaluated the chemopreventive potential of a diet supplemented with natural α-tocopherol-enriched transgenic (TR) Brassica juncea seeds. The modulation of phase I and phase II xenobiotic metabolism and of antioxidative enzymes was compared in the livers of mice fed on a control diet or on a diet supplemented with 2, 4, and 6 % (w/w) of wild-type (WT) or TR seeds. A dose-dependent increase in the specific activities of these enzymes was observed in those animals fed on diet supplemented with TR seeds. In comparison, an increase in the specific activities of antioxidative enzymes was substantial only at higher doses of WT seeds. Consequently, oxidative stress measured in terms of lipid peroxidation and lactate dehydrogenase activity was found to be lower in the case of mice fed with the supplemented diet. However, the chemopreventive potential of TR seeds was more pronounced than that of WT seeds. This study demonstrates the feasibility of fortifying diets with natural α-tocopherol for chemopreventive benefits by means of transgenic manipulation of a commonly used oilseed crop.     

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．     

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.