Effect of nitrogen on thiocyanate content of Brassica oleracea var. acephala leaves

An experiment was carried out to study the effect of nitrogen fertilizer application levels (0, 47, 94 and 188 kg N/ha) on thiocyanate contents of petioles and lamina of two Brassica oleracea L. var. acephala varieties (Thousand-headed kale and Georgia collards).The results showed that N application significantly decreased thiocyanate contents of both laminae and petioles. Applying 47, 94, 188 kg N/ha reduced thiocyanate of laminae and petioles by 26, 41 and 52% and 2, 9 and 39%, respectively. The results also showed that thiocyanate of leaves from Thousand-headed kale was significantly higher than that of Georgia collards. Lamina and petiole thiocyanates of Georgia collards were 79% and 83% of those of Thousand-headed kale, respectively.     

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%.     

羽衣甘蓝SEPALLATA-like基因的系统发育与表达分析

E类MADS-box基因SEPALLATA (SEP)-like在被子植物生殖生长特别是花器官发育方面具有重要作用。为分析羽衣甘蓝E功能MADS-box基因SEP-like基因的序列特征及其在花发育过程中的时空表达模式,以羽衣甘蓝品系‘14 line’为试材,利用cDNA末端快速扩增(Rapid amplification of cDNA ends,RACE)技术克隆了SEP直系同源基因BroaSEP1/2/3 (GenBank登录号:KC967957、KC967958、KC967960)。序列和系统进化树分析表明,这3个基因分别与野甘蓝(Brassica oleracea var. oleracea)、芜菁Brassica rapa、萝卜Raphanus sativus、甘蓝型油菜Brassica napus的SEP1、SEP2、SEP3基因具有很高的同源性。推导的氨基酸序列显示,这些基因编码的蛋白质都包含高度保守的MADS结构域、I结构域和K结构域,每一基因都有其亚家族特异的C-末端功能域SEPⅠ和SEPⅡ基序。BroaSEP1、BroaSEP2、BroaSEP3基因的开放阅读框长...     

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。     

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。     

Anther culture of kale (Brassica oleracea L. convar.acephala (DC.) Alef.)

The pollen development and androgenic ability of 18 kale (Brassica oleracea convar.acephala) genotypes was observed during an anther culture study. Anther culture was successful in 6 of the genotypes and the highest yield obtained was 17 embryos per 100 anthers plated. Two stages of anther development were identified as being responsive to anther culture. The first and most responsive was that corresponding to the late uninucleated stage and the second to the late binucleated stage. These stages correspond with the onset of mitotic events in the microspores. Pollen viability was studied and low viability was noted which declined to zero after 9 days of anther culture. The initial viability level however was not clearly related to androgenic ability. The significance of the production of haploid and dihaploid kale genotypes in the study and breeding of resistance to clubroot is discussed.     

Bottom-up and top-down herbivore regulation mediated by glucosinolates in Brassica oleracea var. acephala

Quantitative differences in plant defence metabolites, such as glucosinolates, may directly affect herbivore preference and performance, and indirectly affect natural enemy pressure. By assessing insect abundance and leaf damage rate, we studied the responses of insect herbivores to six genotypes of Brassica oleracea var. acephala, selected from the same cultivar for having high or low foliar content of sinigrin, glucoiberin and glucobrassicin. We also investigated whether the natural parasitism rate was affected by glucosinolates. Finally, we assessed the relative importance of plant chemistry (bottom-up control) and natural enemy performance (top-down control) in shaping insect abundance, the ratio of generalist/specialist herbivores and levels of leaf damage. We found that high sinigrin content decreased the abundance of the generalist Mamestra brassicae (Lepidoptera, Noctuidae) and the specialist Plutella xylostella (Lepidoptera, Yponomeutidae), but increased the load of the specialist Eurydema ornatum (Hemiptera, Pentatomidae). Plants with high sinigrin content suffered less leaf injury. The specialist Brevicoryne brassicae (Hemiptera, Aphididae) increased in plants with low glucobrassicin content, whereas the specialists Pieris rapae (Lepidoptera, Pieridae), Aleyrodes brassicae (Hemiptera, Aleyrodidae) and Phyllotreta cruciferae (Coleoptera, Chrysomelidae) were not affected by the plant genotype. Parasitism rates of M. brassicae larvae and E. ornatum eggs were affected by plant genotype. The ratio of generalist/specialist herbivores was positively correlated with parasitism rate. Although both top-down and bottom-up forces were seen to be contributing, the key factor in shaping both herbivore performance and parasitism rate was the glucosinolate concentration, which highlights the impact of bottom-up forces on the trophic cascades in crop habitats.     

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。     

NIRS定量分析羽衣甘蓝油含量、蛋白质含量数学模型的建立

应用近红外反射光谱技术(NIRS)采集807份羽衣甘蓝种子光谱数据,然后挑选250份材料,用经典化学方法测试种子油含量和饼粕蛋白质含量.通过偏最小二乘法(PLS)、改良最小二乘法(MPLS)和不同光谱散射及数学处理技术来建立NIRS定量分析数学模型.种子油含量和饼粕蛋白质含量模型的交叉检验相关系数(1-VR)分别为0 9180、0 9062,交叉检验标准差(SECV)分别为1 0606、0 8720,校正标准差(SEC)分别为0 9267、0 8119.两种模型的外部检验相关系数(RSQ)分别为0 949、0 915,相对误差分别小于3 60%、2 70%,预测标准差(SEP)分别为0 881、0 779,检验偏差(BIAS)均较小分别为-0 054、-0 062.研究表明:这两个数学模型的分析结果具有较高的精确度,在品质育种中具有较好的应用前景.     

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.     

The serological specificity of S alleles of homozygous incompatible lines of the marrow-stem kale (Brassica oleracea var.acephala DC.)

The preparation of antisera against S alleles of homozygous incompatible lines of the marrow-stem kale (Brassica oleracea var.acephala DC.) is described. The antisera obtained reacted specifically with homologous antigens on using the serological method of double diffusion into agar. The results confirmed the specificity of S alleles of incompatible lines of the marrow-stem kale.     

激光、高压静电场克服羽衣甘蓝自交不亲和及其分子机制的研究

利用激光、高压静电场对自交不亲和的羽衣甘蓝的花粉进行处理,以期克服其自交不亲和。通过对处理后的花粉的表面结构和蛋白酶活性及其萌发情况的研究,进一步分析激光、高压静电场的生物效应。同时利用分子生物学手段Northern杂交技术,检测了经激光、高压静电场处理的花粉,在授粉后柱头上SLG和SRK基因表达的差异,以探讨激光、高压静电场对生殖反应作用的分子机制。结果表明:通过适宜剂量的激光、高压静电场处理,羽衣甘蓝的花粉萌发率提高,成熟花粉水溶性总蛋白,α-淀粉酶活力,总淀粉酶活力均不同程度的提高,同时自交亲和指数和结籽率提高。从花粉的表面结构的电镜显微观察分析,花粉壁的雕纹受到了伤害,网脊线有轻微断裂,网眼变大。授粉后,取不同时间间隔的柱头样品提取总RNA,利用Northern杂交技术检测SLG和SRK基因的表达量,授粉后24 h激光处理和高压静电场处理与对照比较,SLG和SRK基因的表达量均降低。     

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.     

cDNA cloning and characterization of a new stress-responsive gene BoRS1 from Brassica oleracea var. acephala

A new stress-responsive gene BoRS1 (GenBank accession No. AY373021 ) was isolated from Brassica oleracea var. acephala by rapid amplification of cDNA ends (RACE). The full-length cDNA of BoRS1 was 2076 bp and contained a 1851 bp open reading frame (ORF) encoding 617 amino acids. Sequence analysis indicated that the deduced BoRS1 shared some identities with LTI65 , RD29A, RD29B and COR78 from Arabidopsis thaliana . Southern blot analysis of genomic DNA indicated that other related genes existed and there were two copies of BoRS1 in the genome of B. oleracea . Northern blot analysis revealed that BoRS1 was up-regulated by cold, mannitol, NaCl and abscisic acid (ABA). Expressional fluctuation of time course with ABA implied a two-step induction process. Tissue-specific expression analysis indicated that BoRS1 was expressed in all the tested plant tissues including leaves, stems and roots. Our studies imply that BoRS1 is a new gene that is responsive to environmental stresses such as low temperature, salinity and osmotic stress.     

羽衣甘蓝种子主要品质性状的表现及其相关性

随机挑选148份羽衣甘蓝种质资源和高世代材料,分析了成熟种子的含油量、蛋白质、硫苷和7种主要脂肪酸成分的表现特征及其相关性。结果表明:羽衣甘蓝成熟种子平均含油量为29.48%,平均蛋白质含量为45.13%,含油量和蛋白质总量为74.61%。硫苷含量的变幅最大,变异系数为31.72%。7种主要脂肪酸成分中,油酸和芥酸的含量较高,其次为亚油酸,棕榈酸和硬脂酸的含量较低。除硫苷含量和硬脂酸含量外,其余9个性状的表现均呈单峰正态分布。相关性分析表明,大多数性状间都具有显著或极显著的相关性,这与对甘蓝型、白菜型和芥菜型3种类型油菜的研究结果相一致。在羽衣甘蓝中存在一些优异的种质资源,通过筛选可以在油菜优质育种中加以利用。     

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.     

观赏羽衣甘蓝SSR标记分型与亲缘关系研究

观赏羽衣甘蓝凭借优良的观赏特性和抗逆性已经成为重要的冷季观赏植物。国内观赏羽衣甘蓝育种起步较晚,并且缺乏对种质资源遗传背景的系统研究。本研究应用SSR标记对不同类型的观赏羽衣甘蓝材料进行标记分型和亲缘关系分析。从99对均匀分布于甘蓝基因组的SSR引物中筛选出46对多态性好的引物,对27份不同类型的观赏羽衣甘蓝材料进行标记分型,共扩增出210个多态性位点,平均PIC值为0.58。进一步利用标记分型结果进行STRUCTURE群体结构、UPGMA聚类和聚类热图分析,结果显示3种分析结果基本一致,可以将27份材料分为圆叶、羽叶和皱叶3种类型,其中圆叶和羽叶类型的亲缘关系更近,与皱叶类型的亲缘关系较远;STRUCTURE分析还可以将双亲为不同类型的杂交种材料进行区分;聚类热图分析可以将标记分型结果形象的展示出来。本研究为进一步建立观赏羽衣甘蓝分子指纹图谱,明确种质资源的遗传背景,建立观赏羽衣甘蓝分子标记辅助选择育种体系,培育具有自主知识产权的新品种奠定基础。     

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。     

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.