The characteristic high sulfate content in Brassica oleracea is controlled by the expression and activity of sulfate transporters

The uptake and distribution of sulfate in BRASSICA OLERACEA, a species characterised by its high sulfate content in root and shoot, are coordinated and adjusted to the sulfur requirement for growth, even at external sulfate concentrations close to the K (m) value of the high-affinity sulfate transporters. Plants were able to grow normally and maintain a high sulfur content when grown at 5 or 10 microM sulfate in the root environment. Abundance of mRNAs for the high affinity sulfate transporters, BolSultr1;1 and BolSultr1;2, were enhanced at 相似文献   

甘蓝型油菜小孢子培养技术的几项改进

本研究在NLN-16和NLN-13的培养基中分别加入0.1mg/L 6-BA和0.05%的活性碳,结果表明6-BA对小孢子再生胚有明显地促进作用,再生胚的频率比对照增加26枚/皿,经分析达到显著水平;而0.05%的活性碳对小孢子再生胚促进作用不显著。对甘蓝型油菜小孢子培养再生植株的染色体加倍及移栽的研究结果表明,在小孢子培养初期加50mg/L秋水仙碱加倍效率最佳,加倍率达到67.6%。小孢子培养的再生苗移栽至大田后,采用遮阳网覆盖小苗,移栽成活率达到87.6%。 Abstract:The application of microspore culture technique was restricted because of its low frequency of embryogenesis and chromosome doubling.Two methods of enhancing the frequency of embryogenesis were employed in the study,namely,activated charcoal treatment in NLN-13 media and 6-BA treatment in NLN-16 media.The treatment with 0.05% activated charcoal produced 24 embryos per plate,which increased 1.7 embryos per plate,as compared with the treatment without activated charcoal.However,the analysis of T-test showed that it was not significant.After adding 0.1mg/L 6-BA in NLN-16 media,the frequency of embryogeny was 38.3 embryos per plate,and it was 26 embryos more per plate than that of CK.Analysis of T-test is significant.This indicates that 6-BA promotes embryogeny in microspore culture.Adding 50mg/L colchicines in NLN-16 media,the doubling frequency was 67.6%.The plantlets transplanted into field with two methods of light-covered net and plastic films were investigated.A survival rate of 87.6% was obtained using light-covered method whereas 57.7% survived using plastic film method.     

甘蓝型油菜矮秆突变材料99CDAM的发现及遗传分析

随着杂种优势的利用,油菜株高增加了20cm以上,导致油菜生长后期倒伏的风险加大。通过利用特殊矮秆基因来控制株高将是解决倒伏问题的有效方法。在甘蓝型油菜自交多代品系中发现了一个株高85cm左右的矮秆突变株99CDAM,该突变株具有开花早、分枝多等优良特性,产量性状和品质性状较好,各性状都能稳定遗传,具有重要的育种价值。对99CDAM和高秆品系2091、7045和7350的正反交F1以及99CDAM和2091的F2、BC1及F2:3代的遗传分析结果表明： 99CDAM的矮秆性状受3对隐性基因控制,存在母体细胞质效应,与以往的甘蓝型矮秆油菜有明显的区别。     

Conservation of S-locus for self-incompatibility in Brassica napus (L.) and Brassica oleracea (L.)

 Self-incompatibility (SI) in Brassica is a sporophytic system, genetically determined by alleles at the S-locus, which prevents self-fertilization and encourages outbreeding. This system occurs naturally in diploid Brassica species but is introduced into amphidiploid Brassica species by interspecific breeding, so that in both cases there is a potential for yield increase due to heterosis and the combination of desirable characteristics from both parental lines. Using a polymerase chain reaction (PCR) based analysis specific for the alleles of the SLG (S-locus glycoprotein gene) located on the S-locus, we genetically mapped the S-locus of B. oleracea for SI using a F₂ population from a cross between a rapid-cycling B. oleracea line (CrGC-85) and a cabbage line (86-16-5). The linkage map contained both RFLP (restriction fragment length polymorphism) and RAPD (random amplified polymorphic DNA) markers. Similarly, the S-loci were mapped in B. napus using two different crosses (91-SN-5263×87-DHS-002; 90-DHW-1855-4×87-DHS-002) where the common male parent was self-compatible, while the S-alleles introgressed in the two different SI female parents had not been characterized. The linkage group with the S-locus in B. oleracea showed remarkable homology to the corresponding linkage group in B. napus except that in the latter there was an additional locus present, which might have been introgressed from B. rapa. The S-allele in the rapid-cycling Brassica was identified as the S₂₉ allele, the S-allele of the cabbage was the S ₅ allele. These same alleles were present in our two B. napus SI lines, but there was evidence that it might not be the active or major SI allele that caused self-incompatibility in these two B. napus crosses. Received: 7 June 1996/Accepted: 6 September 1996     

油菜（Brassica napus L.）种子脱水耐性获得过程中肌醇半乳糖苷合成酶活性与可溶性糖含量的变化

在这个研究中测量不同发育时期的油菜种子中可溶性糖含量与肌醇半乳糖苷合成酶（galactinol synthase,GOLS）活性,将二者的变化趋势与种子脱水耐性获得的过程相比较并对结果进行相关性分析。结果显示油菜种子脱水耐性获得过程中,葡萄糖和果糖含量均随着发育期的延长而下降,蔗糖则保持较高水平;肌醇含量下降而肌醇半乳糖苷含量上升;棉子糖系列寡糖（raffinose familyolig osaccharides,RFO）含量随着种子发育而上升,特别是水苏糖,在成熟种子中可以达到相当高的浓度。油菜种子发育中期,细胞内GOLS活性开始上升,至贮藏物积累完成时达到最大。GOLS活性变化与种子肌醇半乳糖苷积累速度、RFO含量及种子的脱水耐性呈一定的正相关关系。我们认为GOLS促使RFO积累,从而对种子脱水耐性的获得产生重要影响。     

Intergeneric Somatic Hybridization Between Brassica napus L. and Sinapis alba L.

Electrically induced protoplast fusion was used to produce somatic hybrids between Brassica napus L. and Sinapis alba L. Seven hybrids were obtained and verified by the simple sequence repeat and cleaved amplified polymorphic sequence analysis of the genefael, indicating that the characteristic bands from S. alba were present in the hybrids. The hybridity was also confirmed by chromosome number counting because the hybrids possessed 62 chromosomes, corresponding to the sum of fusion-parent chromosomes. Chromosome pairing at meiosis was predominantly normal, which led to high pollen fertility,ranging from 66% to 77%. All hybrids were grown to full maturity and could be fertilized and set seed after self-pollination or back-crosses with B. napus. The morphology of the hybrids resembled characteristics from both parental species. An analysis of the fatty acid composition in the seeds of F1 plants was conducted and the seeds were found to contain different amounts of erucic acid, ranging from 11.0% to 52.1%.     

油菜种子发育早期的油体发生与调控

以甘蓝型油菜( Brassica napus L.)品种‘Westar’和‘Topas’为材料,通过超微结构观察和荧光定量PCR技术对油菜胚胎发育早期油体的发生、油体蛋白及脂肪酸合成转录因子基因的表达情况进行分析。结果显示:油体出现在油菜胚胎发育早期,在授粉9 ~ 11 d后(球形胚时期)的胚体和胚柄中均存在直径小于0. 5 μm的油体;荧光定量实验结果表明,除 BnCLO3 的表达量在整个胚胎发育阶段无明显变化外,其他油体蛋白基因 Oleosins 、 Steroleosins 和 BnCLO1 的表达量在心形胚时期就明显增多并持续增长;脂肪酸合成转录因子 BnLEC1 、 BnL1L 、 BnWRI1 和 BnFUS3 在胚胎发育阶段,基因表达规律均呈先上升再下降的趋势,但达到最高值的时间存在差异,其中 BnLEC1 最早, BnL1L 其次, BnWRI1 和 BnFUS3 较晚。研究结果表明甘蓝型油菜在球形胚时期出现油体,其结构蛋白和转录调控因子基因的表达自心形胚开始明显增多。     

油菜种子发育早期的油体发生与调控

以甘蓝型油菜（Brassica napus L.）品种‘Westar’和‘Topas’为材料,通过超微结构观察和荧光定量PCR技术对油菜胚胎发育早期油体的发生、油体蛋白及脂肪酸合成转录因子基因的表达情况进行分析。结果显示：油体出现在油菜胚胎发育早期,在授粉9~11 d后（球形胚时期）的胚体和胚柄中均存在直径小于0.5 μm的油体;荧光定量实验结果表明,除BnCLO3的表达量在整个胚胎发育阶段无明显变化外,其他油体蛋白基因Oleosins、Steroleosins和BnCLO1的表达量在心形胚时期就明显增多并持续增长;脂肪酸合成转录因子BnLEC1、BnL1L、BnWRI1和BnFUS3在胚胎发育阶段,基因表达规律均呈先上升再下降的趋势,但达到最高值的时间存在差异,其中BnLEC1最早,BnL1L其次,BnWRI1和BnFUS3较晚。研究结果表明甘蓝型油菜在球形胚时期出现油体,其结构蛋白和转录调控因子基因的表达自心形胚开始明显增多。     

Aberrant gene expression in the Arabidopsis SULTR1;2 mutants suggests a possible regulatory role for this sulfate transporter in response to sulfur nutrient status

Sulfur is required for the biosynthesis of cysteine, methionine and numerous other metabolites, and thus is critical for cellular metabolism and various growth and developmental processes. Plants are able to sense their physiological state with respect to sulfur availability, but the sensor remains to be identified. Here we report the isolation and characterization of two novel allelic mutants of Arabidopsis thaliana, sel1‐15 and sel1‐16, which show increased expression of a sulfur deficiency‐activated gene β‐glucosidase 28 (BGLU28). The mutants, which represent two different missense alleles of SULTR1;2, which encodes a high‐affinity sulfate transporter, are defective in sulfate transport and as a result have a lower cellular sulfate level. However, when treated with a very high dose of sulfate, sel1‐15 and sel1‐16 accumulated similar amounts of internal sulfate and its metabolite glutathione (GSH) to wild‐type, but showed higher expression of BGLU28 and other sulfur deficiency‐activated genes than wild‐type. Reduced sensitivity to inhibition of gene expression was also observed in the sel1 mutants when fed with the sulfate metabolites Cys and GSH. In addition, a SULTR1;2 knockout allele also exhibits reduced inhibition in response to sulfate, Cys and GSH, consistent with the phenotype of sel1‐15 and sel1‐16. Taken together, the genetic evidence suggests that, in addition to its known function as a high‐affinity sulfate transporter, SULTR1;2 may have a regulatory role in response to sulfur nutrient status. The possibility that SULTR1;2 may function as a sensor of sulfur status or a component of a sulfur sensory mechanism is discussed.     

EMS诱变甘蓝型油菜M_2代群体的表型突变研究

利用4种浓度EMS处理甘蓝型油菜NJ7982种子,选取其中诱变效果好的处理(0.4%EMS),研究其后代突变型和突变频率。对4.8万株的M2代群体鉴定结果表明,6种器官性状在群体中均出现了突变。子叶突变性状包括3子叶、子叶黄化等,占群体的0.22%;叶片突变性状包括黄化叶、白化叶、紫色叶、上卷叶、下卷叶等,占0.74%;花器突变性状包括紫色花蕾、死蕾、3花瓣、6花瓣、白色瓣、花瓣黄白镶嵌、花瓣皱缩、完全不育、部分可育等,占9.38%;株型突变性状包括矮秆、紫茎等,占4.98%;角果突变性状包括粗角、长角、紫角等,占2.79%;种皮黄色,占0.40%;总的表型突变频率为18.51%。这些遗传多样性的突变材料,为甘蓝型油菜种质创新及品种遗传改良提供基础性材料。     

Changes in protein and carbohydrate content during development of seeds and siliquas of rapeseed (Brassica napus L.)

Abstract

A study was made on the changes observed in the protein, starch and soluble sugar content during development of siliquas and seeds of rapeseed grown in central Italy.

Concentration of starch and soluble sugars in the seed increases to 75 per cent dry matter during the first few weeks of pod development and then drops to minimum values. The protein increases steadily until maturity, when a level of 0.85 mg per seed is reached, equivalent to 18 per cent dry matter. The protein and starch in the hull? decrease continuously during development, while in the initial stages the soluble sugars are accumulated until they account for 33 per cent dry matter, after which they decline towards maturity.     

单核苷酸多态性及其在油菜分子标记辅助育种中的应用

油菜是一种重要的油料作物,油菜的基因组功能研究已引起了各国政府和科研工作者的高度重视。SNP是目前为止分布最广泛,存在数量最多且标记密度最高的一种遗传多态性标记。对油菜等作物的基因组功能研究提供了丰富的信息资源。综述了近来单核苷酸多态性的研究进展以及其在油菜的分子标记辅助品质育种中的可能用途。     

Cytochemical Localization of Phenolic Compounds in Columella Cells of the Root Cap during Maturation of Seeds of Brassica napus L.

Abstract: Ultrastructural localization of phenolic compounds in the columella of the root cap of a canola embryo in Brassica napus L. during subsequent seed maturation stages was investigated. In bright green seeds the first signs of maturation and appearance of the first phenolic structures in the cell nuclei and cytoplasm in the area of the initial centre were found. Phenolics spread acropetally, gradually covering the whole columella in the dark green seeds. An increase in the number of phenolics occurred, together with an increase in the number of endoplasmic reticulum structures and in storage materials. In brown seeds, together with ultrastructural degradation and the end of production of storage substances, the number of phenolic deposits in the nucleus and cytoplasm decreases until their disappearance in black dormant seeds. Simultaneously, numerous phenolic deposits appear between the plasmalemma and the cell walls in the outer layer of the cap columella.     

Induction of a specific isoenzyme of glutamate dehydrogenase during isolation and the first 48 h of culture of Brassica napus leaf protoplasts

The specific activity of NADH‐glutamate dehydrogenase (GDH, EC 1.4.1.2) in leaf protoplasts ( Brassica napus L. cv. Bronowski) was initially low and progressively increased during culture in Murashige and Skoog (MS) medium and MS (−NH₄) (ammonium nitrate‐free MS) medium in the dark. Native polyacrylamide gel electrophoresis (PAGE) and tetrazolium staining revealed that the high specific activity of NAD‐GDH (deamination) in leaves correlated with the cathodal isoenzymes, and the high specific activity of NADH‐GDH (amination) in leaf protoplasts to the anodal ones. Changes in isoenzyme pattern were correlated with an increase in the specific activity of NADH‐GDH but not with the NADH‐GDH/NAD‐GDH ratio. The increase in NADH‐GDH (amination) activity of leaf protoplasts was correlated with the occurrence of the isoenzyme GDH7, which was not detected in leaves.     

油菜BnMKK4全长基因的克隆及表达分析

从‘陇油6号’油菜中克隆得到了一个新的BnMKK4基因的cDNA,全长1317bp,其中包括993bp的开放阅读框,159bp的5’非翻译区（5^1UTR）,165bp的3。非翻译区（3’UTR）。与拟南芥AtMKK4有很高的同源性,因此命名为BnMKK4（GenBank登录号JF268686）。该基因编码330个氨基酸的蛋白质,分子量36．5kDa,等电点为9．01。实时荧光定量PCR结果显示该基因表达受低温胁迫和盐胁迫的诱导,表明该基因在油菜适应低温胁迫和盐胁迫的过程中发挥作用。