羽衣甘蓝授粉过程中柱头蛋白质的泛素化

分析自花授粉与异花授粉后的羽衣甘蓝柱头蛋白质泛素化变化的结果表明,自花授粉30min后的柱头蛋白质泛素化水平显著增加,45min时达到峰值;异花授粉的柱头蛋白质泛素化水平没有变化。     

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.     

Two novel bioactive glucosinolates from Broccoli (Brassica oleracea L. var. italica) florets

Two novel glucosinolates along with one known glucosinolate were isolated from Broccoli (Brassica oleracea L. var. italica) florets. Their structures were established mainly by 1D ((1)H and (13)C NMR), 2D NMR ((1)H-(1)H COSY, DEPT 135°, HSQC and HMBC), and Tandem MS-MS spectrometric data as 2-mercaptomethyl sulfinyl glucosinolate [(Z)-4-(methylsulfinyl)-N-(sulfooxy)-2-((2'S,3'R,4'S,5'S,6'R)-3',4',5'-trihydroxy-6'(hydroxylmethyl)-2'-mercapto tetrahydro-2H-pyran-2-yl) butane amide] 1, (Z)-1-((2S,5S)-5-hydroxytetra-hydro-2H-pyran-2-ylthio)-2-(1H-indol-3-yl) ethylidene amino sulfate 2 and a known cinnamoyl [6'-O-trans-(4″-hydroxy cinnamoyl)4-(methylsulphinyl)butyl glucosinolate] 3. Compound 1 exhibited scavenging activity against DPPH with an inhibitory concentration IC(50) of 20mM, whereas compound 3 was a weak antioxidant when compared to the standard quercetin (5mM) as a positive control. Both the compounds showed a significant and similar antimicrobial activity against Staphylococcus aureus with an IC(50) of <625μg/mL when compared to antibiotic duricef. Against Salmonella typhimurium the IC(50) of 1 and 3 was determined as <625μg/mL and <1250μg/mL, respectively, when compared to ampicillin (IC(50) ?39μg/mL) as a positive control.     

羽衣甘蓝花粉母细胞减数分裂及雄配子体发育观察

采用常规压片法对羽衣甘蓝花粉母细胞减数分裂及雄配子体发育进行了细胞学观察,结果显示:羽衣甘蓝减数分裂类似甘蓝种,细胞质分裂为同时型,四分体以正四面体型或十字交叉型为主;终变期有9个二价体,此时可进行染色体计数;中期Ⅰ和Ⅱ少数细胞中可见赤道板外染色体,后期Ⅰ和Ⅱ存在落后染色体,四分体时期可观察到少量含微核的异常四分体;单核靠边期时花蕾长度约为2.0~2.2 mm,小孢子经过发育最终成为3-细胞型花粉并具3个萌发孔,成熟花粉中败育花粉比率为1.3%.     

Comparative analysis of glucosinolate production in hairy roots of green and red kale (Brassica oleracea var. acephala)

Abstract

Glucosinolates (GSLs) are sulfur- and nitrogen-containing secondary metabolites that function in plant defense and provide benefits to human health. In this study, using Agrobacterium rhizogenes R1000, green and red kale hairy roots were established. The expression levels of GSLs biosynthesis genes and their accumulation in both kale hairy roots were analyzed by quantitative real-time PCR and HPLC. The results showed that the expression of most indolic GSLs biosynthesis genes was higher in the hairy roots of green kale than in that of red kale. In contrast, the expression of BoCYP83A1 and BoSUR1 encoding key enzymes aromatic GSL biosynthesis was significantly higher in red kale hairy root. The HPLC analysis identified six GSLs. The levels of 4-methoxyglucobrassicin, glucobrassicin, and 4-hydroxyglucobrassicin were 6.21, 5.98, and 2 times higher, respectively, in green kale than in red kale, whereas the levels of neoglucobrassicin and gluconasturtiin were 16.2 and 3.48 times higher, respectively, in red kale than in green kale. Our study provides insights into the underlying mechanisms of GSLs biosynthesis in kale hairy roots and can be potentially used as “biological factories” for producing bioactive substances such as GSLs.     

Die Wirkung einer Vorbestrahlung auf den Wuchsstoffgehalt von X-2-Pflanzen bei Brassica oleracea var. acephala Von

The Effect of Pre-irradiation on the Content of Growth Substances in Xi-Plants of Brassica oleracea var. acephala. The auxin content of growth stimulated young plants (Brassica oleraceav ar. acephala) of the X2-generation, originating from X-ray irradiated seeds (75 kR) was determined. Methanolic extracts were separated on thin-layer-chromatograms, which were analyzed by ultra-violet light and various chemical reactions. Quantitative determinations of growth substances were made using the Avena-coleoptile-cylinder-test. The following results were obtained: 1. X2-plants had a higher content of IAN compared with unirradiated control samples; 2. IAAm was detectable in only two out of 40 experiments; 3. Two fluorescent substances were detected: a hydroxycinnarmc acid derivative and a long chained unsatu-rated ketone or aldehyde, which is still unknown.     

Genetics of aliphatic glucosinolates. IV. Side-chain modification in Brassica oleracea

The biochemical and genetical relationship between aliphatic glucosinolates which have methylthioalkyl, methylsulphinylalkyl and alkenyl side chains has not been resolved by biochemical studies. In this study, two hypothetical models are tested by the genetic analysis of a backcross population between Brassica drepanensis and B. atlantica. The results support one of the models in which 3-methylthiopropyl glucosinolate is sequentially converted to 3-methylsulphinylpropyl, and then to 2-propenyl glucosinolate, by the action of dominant alleles at two loci. RFLP mapping positioned both loci on the same linkage group homologous to the B. napus N19 linkage group. The implication of the results for the genetic manipulation of glucosinolates in Brassica to improve flavour and nutritional properties, and in order to investigate plant-insect interactions, is discussed.     

Effect of hexavalent chromium on the growth and physiological and biochemical parameters on Brassica oleracea L. var. acephala DC

In order to determine the toxic effect of chromium Cr(VI) on the seed germination, the root and shoot length, the root-cotyledonary leaves, the fresh and dry weight in eight-day-old seedlings Brassica oleracea L. var. acephala DC (kale) were treated with various concentrations of Cr in the growth medium. The accumulation of chromium in the tissues was determined in the cotyledons and the roots of the kale seedlings. High rate of Cr uptake was observed in the roots. But the organs could not accumulate large amount Cr. The effect of Cr on B. oleracea var. acephala was evaluated by changes in chlorophyll a, b, lipid peroxidation, proline, ascorbate, protein carbonyl groups, non-protein thiols and peroxidase activity. There were significant decreases in chlorophylls a, b content of the plants treated with Cr. Chromium treated kale seedlings had higher lipid peroxidation and the protein carbonyl groups in cotyledonary leaves than the roots. The changes refer to toxic effects of Cr. There were increases in the non-protein thiol, the total ascorbate, and proline content in the cotyledons and the roots of the seedlings grown on the media containing 0.1 and 0.15 mM Cr. The guaiacol peroxidase activity was higher in the roots of the seedlings than their cotyledons.     

Agrobacterium rhizogenes-mediated transformation of Brassica oleracea var. sabauda and B. oleracea var. capitata

Agrobacterium rhizogenes A4M70GUS-mediated transformation of Savoy cabbage (Brassica oleracea L. var. sabauda) and two local lines of cabbage (B. oleracea L. var. capitata) was obtained using hypocotyl and cotyledon explants. The percentage of explants which formed roots was very high in all genotypes: 92.3 % in Savoy Gg-1, 64.4 % in cabbage P₂₂I₅, and 87.2 % in P₃₄I₅. Spontaneous shoot regeneration of excised root cultures grown on the hormone-free medium occurred in all three genotypes. In cabbage lines P₂₂I₅ and P₃₄I₅ shoot regeneration was higher (9.3 and 2.6 % respectively) than in Savoy cabbage Gg-1 (1.3 %). Transgenic nature of hairy root-derived plants was evaluated by GUS histological test and PCR analysis. All the tested cabbage shoots were GUS positive whilst in a Savoy cabbage GUS expression was registered only in 55 % of tested clones. PCR analysis demonstrated the presence of the GUS gene in regenerated shoot clones and in T₁ progeny.     

Estimation of genetic diversity Among Turkish kale populations (Brassica oleracea var. acephala L.) using RAPD markers

20 populations of kale (B. oleracea var. acephala L.) selected from 127 populations for fresh consumption terms of yield and leaf quality characteristics as superior types using weight-based ranking method from the Black Sea Region of Turkey were evaluated at the DNA level using randomly amplified polymorphic DNA (RAPD) markers compared to some morphological characters. The 7 primers selected from 100 decamers used generated 110 bands, of which 60 (54.5%) were polymorphic. Jaccard's genetic distances were calculated and dendogram was generated using the UPGMA algorithm. The dendogram obtained were classified into three main groups and four subgroups. The accessions showed a limited clustering in compare to morphological characters such as the number of leaf, leaf intentation of the margin, leaf and midrib color and thickness of midrib than geographical characteristics. Leaf color and midrib thickness characters clustered in the same group as OR49 and G18 accessions; S20, G6 and OR37 accessions, respectively.     

几种影响羽衣甘蓝小孢子胚状体成苗的因素

羽衣甘蓝成熟小孢子胚转到固体培养基上可直接萌发成苗。成苗率与基因型、培养基成分和培养温度有关。MS+1.0%琼脂+3%蔗糖是适宜的成苗培养基;100MG·L-1活性炭对鱼雷形胚成苗起促进作用;10℃低温培养10D可提高成苗率。     

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.     

羽衣甘蓝胁迫应答基因BoRS1转化烟草的初步研究

将克隆于羽衣甘蓝的胁迫应答基因BoRS1连入中间载体p35S-2300：：gus：：noster相应位点,成功地构建了含BoRS1基因的植物双元表达载体p35S-2300：：BoRS1：：noster,并通过农杆菌介导法对烟草进行了遗传转化。PCR检测结果表明目的基因BoRS1已成功地导入并整合到烟草基因组中。RT-PCR分析显示,在不同的转基因烟草植株中BoRS1表达量存在差异。转BoRS1烟草的耐干性和甘露醇胁迫研究表明,BoRS1基因的表达对提高植物抗干旱胁迫能力有一定的作用。     

Bottom-up and top-down regulation of decomposition in a tropical forest

The soil nutrients, microbes, and arthropods of tropical forests are patchy at multiple scales. We asked how these three factors interact to generate patterns of decomposition in 450 100 cm² litterbags arrayed along a 50 m ridge top in a Panama rainforest. We tested top-down (via grazing by microbivores like collembola and diplopods) and bottom-up (via added N and P) effects on the decomposition of cellulose. By using a 1,000-fold gradient in mesh size we generated a two-fold gradient in arthropod grazing. Microbivore grazing first retarded then ultimately enhanced decomposition rates. Micropulses of N and P (simulating concentrated urine) enhanced neither decomposition rates nor microbivores but increased the abundance of predacious ants. Decomposition rates also varied across the ridge, and were lowest in a plot with the deepest litter and highest soil moisture. These data generate the working hypothesis that N and P cascade upward at grains of 100 cm² to enhance a major predator in the litter; predators then absorb any increases in microbivores attracted to the extra fungal growth. These population interactions are in turn embedded in mesoscale variability generated by individual tree canopies that drive changes in litter quality and soil moisture. Electronic supplementary material The online version of this article (doi:) contains supplementary material, which is available to authorized users.     

羽衣甘蓝的组织培养

1　植物名称　羽衣甘蓝 (Ｂｒａｓｓｉｃａｏｌｅｒａｃｅａｖａｒ.Ｒｕｂｒａ)。2　材料类别　无菌苗幼嫩茎段。3　培养条件　 (1 )种子萌发培养基 :1 2ＭＳ ＧＡ3 1ｍｇ·Ｌ-1(单位下同 ) ;(2 )增殖培养基 :ＭＳ 6 ＢＡ 2 .0 ＮＡＡ 0 .1 ;(3 )生根培养基 :1 2ＭＳ ＮＡＡ 0 .0 5。所有培养基均加 0 .7%琼脂和 3 %的蔗糖 ,ｐＨ 5 .8,培养温度 (2 5± 2 )℃ ,光照 1 2ｈ·ｄ-1,光照度 2 0 0 0ｌｘ左右。4　生长与分化情况4.1　无菌材料的获得　种子用自来水洗 3次 ,再用 75 %酒精消毒 3 0ｓ,ＨｇＣｌ2 消毒 8ｍｉｎ ,再用无菌水洗 3…     

结球甘蓝和青花菜小孢子胚植株再生

结球甘蓝（Brassica oleracea var.capitata）和青花菜（Brassica oleracea var.italica）小孢子胚再生植株频率低是目前影响游离小孢子培养技术有效应用的关键问题之一,研究其小孢子胚植株再生频率的影响因素,提高胚再生植株频率,对促进游离小孢子培养技术在甘蓝类蔬菜育种中更好地应用具有重要意义。该文以结球甘蓝中甘11和青花菜TI-111等基因型为试材,对影响游离小孢子胚再生成植株的固体培养基类型、琼脂浓度、胚的类型及胚在液体培养基中的滞留时间等因素进行了研究。结果表明：游离小孢子培养25天的子叶胚在琼脂浓度为1%–1.25%的B5培养基上植株再生频率最高。进一步通过8个不同基因型对上述实验结果进行了验证,结果显示,游离小孢子培养25天的子叶胚在1%琼脂浓度的B5培养基上植株再生频率达77.8%–97.2%。     

基于转录组测序的羽衣甘蓝叶色相关基因分析

叶色是羽衣甘蓝重要的观赏性状之一。本研究采用Illumina Hi-Seq2500高通量测序技术,对基因型纯合的紫叶和白叶羽衣甘蓝叶片进行转录组测序,筛选差异基因并与GO和KEGG数据库比对进行注释分析,分析羽衣甘蓝叶色形成相关基因。结果显示,获得高质量短读序共104 608 770条,筛选出紫叶相对白叶的差异表达基因1 993个,其中上调表达基因1 094个,下调表达基因899个。根据GO功能分类可分为生物过程、细胞组分和分子功能3大类64功能组。根据KEGG代谢通路分析可以分为171类,在叶色相关的类黄酮生物合成途径中黄酮醇合成酶(FLS)、二氢黄酮醇4-还原酶(DFR)上调以及类胡萝卜素生物合成途径中的类胡萝卜素β-环化酶上调与紫叶形成关系密切。本研究丰富了羽衣甘蓝的转录组信息,获得了一些差异表达基因,为进一步研究羽衣甘蓝叶色形成的遗传机制提供了有价值的信息。     

羽衣甘蓝柱头酵母双杂交cDNA文库的构建与分析

目的:构建应用于酵母双杂交系统的羽衣甘蓝柱头cDNA文库。方法:以羽衣甘蓝S13-bS13-b自交不亲和系为材料,提取柱头的总RNA,用亲和层析法分离纯化mRNA,利用CytoTrapXR建库试剂盒构建羽衣甘蓝柱头cDNA文库。结果:羽衣甘蓝柱头cDNA原始文库的库容量为2.5×105;扩增后文库的库容量约为4×108,重组率为96%,插入片段大小为0.4～3kb,平均长度在0.8kb左右。结论:构建了应用于酵母双杂交系统的羽衣甘蓝自交不亲和系柱头的cDNA文库,为探讨芸苔属植物自交不亲和的分子机理奠定了基础。     

Growth and shoot formation in protoplast-derived calli of Brassica oleracea ssp. acephala and ssp. capitata

The effect of media components and environmental factors on growth and organogenesis of protoplast-derived calli of curly kale and cabbage were tested. Optimal growth (fresh weight increase of calli, shoots and roots) was found at 60 mM sucrose. Lower sucrose concentrations (3–30 mM) were favourable for shoot formation. Nitrate concentrations from 23 to 100 mM in combination with 8 or 21 mM ammonium were optimal for shoot formation. However, growth was reduced by high (100 mM) nitrate concentration. The effects of various organic nitrogen compounds at 0.5 and 2 mM were tested. Glutamine did not influence shoot formation and barely growth. Proline at 0.5 mM stimulated growth of cabbage calli but decreased growth of curly kale calli, and at 2 mM, proline also inhibited shoot production. Adenine sulphate decreased growth of cabbage calli at 0.5 mM, and at 2 mM shoot production was also reduced. Spermidine and spermine inhibited both growth and differentiation. Putrescine resulted in about 50% higher fresh weights, and also increased the number of calli producing shoots by about 35%. More calli produced shoots in white light than in blue or red light or in darkness. The length of the photoperiod or intensity of light was not critical for shoot production.