甘蓝型油菜和白菜型油菜种间杂种的小孢子培养

利用分离小孢子培养从甘蓝型油菜（Ｂｒａｓｓｉｃａ ｎａｐｕｓ）和白菜型油菜（Ｂ． ｃａｍ ｐｅｓｔｒｉｓ）种间杂种中获得了胚和再生植株。所用的培养程序是,将甘蓝型油菜和白菜型油菜杂种小孢子在蔗糖浓度为１７％ 、ＢＡ 为０．１ ｍ ｇ／Ｌ的液体ＮＬＮ 培养基中３２ ℃下暗培养４８ ｈ,再转入蔗糖浓度为１０％ 的ＮＬＮ 培养液中２５ ℃下暗培养３ 周。不同杂种间小孢子胚胎发生能力存在差异,其中ＵＭ９２１（白菜型油菜）×９１１１８６（甘蓝型油菜）正反交杂种的胚产量显著高于供试的其它组合。供体植株种植在１０ ℃／５ ℃（昼／夜）条件下能显著改善杂种小孢子胚产量和质量。杂种小孢子胚产量和杂种植株每荚种子数存在极显著正相关,但杂种植株的花粉育性和胚产量间相关不显著。大多数甘蓝型油菜和白菜型油菜杂种小孢子胚衍生植株为非整倍体,２２．８％ 的植株起源于具亲本染色体数的小孢子,几乎全部为ｎ＝ １９ 的类型。讨论了影响种间杂种小孢子胚胎发生的因素以及种间杂种小孢子培养技术的可能用途     

芸苔属花粉—下胚轴原生质体融合再生杂种小植株

从青菜（Ｂｒａｓｓｉｃａ ｃｈｉｎｅｎｓｉｓＬ．）单胞中后期至二胞早期花粉分离出原生质体,用聚乙二醇法诱导其与甘蓝型油菜（Ｂ．ｎａｐｕｓＬ．）下胚轴原生质体融合。通过控制双亲原生质体的数量与比率,提高了异源融合率。融合体在离体条件下发生细胞分裂,形成愈伤组织,再生了小植株。染色体计数与酯酶同工酶酶谱分析初步证明获得了１ 株异源三倍体,２ 株异源四倍体。这是以游离花粉时期的原生质体与体细胞原生质体融合,取得“配子－体细胞杂交”成功的首次报道     

抗芜菁花叶病毒转基因甘蓝型油菜的研究

以子叶柄为材料,建立了甘蓝型油菜（ＢｒａｓｓｉｃａｎａｐｕｓＬ．）双低品种的再生体系。通过子叶柄与农杆菌（ＡｇｒｏｂａｃｔｅｒｉｕｍｔｕｍｅｆａｃｉｅｎｓＬＢＡ４４０４）共培养,将表达载体ｐＢＴｕ中芜菁花叶病毒外壳蛋白（ＴｕＭＶ－ＣＰ）基因以整合方式导入甘蓝型油菜,然后用卡那霉素进行筛选,获得了油菜再生植株。经ＰＣＲ特异性扩增、点杂交和Ｓｏｕｔｈｅｒｎ印迹分析,证明再生植株基因组ＤＮＡ中整合了ＴｕＭＶ－ＣＰ基因。攻毒实验表明,有ＴｕＭＶ－ＣＰ基因插入的工程油菜对ＴｕＭＶ均有不同程度的抗性。     

油菜叶绿体核糖体蛋白基因rps7的克隆和序列分析

从甘蓝型油菜（Ｂｒａｓｓｉｃａ ｎａｐｕｓ ｃｖ．Ｈ１６５）叶绿体基因组克隆得到了编码核糖体蛋白的基因ｒｐｓ７。经序列分析得知，该基因编码区包含个核苷酸，编码一个分子量为２０ １０９ Ｄ、由１５５个氨基酸组成的蛋质。该基因的核苷酸和编码的氨基酸序列与烟草对应基因的同源性皆高达９７％；而与水稻对应基因的同源性为９０％和８４％。该基因不含内含子，没有典型的ＳＤ序列，但在５’端－２５～－２２位发现一个与     

根癌农杆菌对甘蓝型油菜的转化及转基因植株的再生

用根癌农杆菌（Ａｇｒｏｂａｃｔｅｒｉｕｍ ｔｕｍ ｅｆａｃｉｅｎｓ）共培养法把外源基因导入甘蓝型油菜（Ｂｒａｓｓｉ－ｃａ ｎａｐｕｓＬ．）主要栽培品种“云北２ 号”,获得转基因植株。所用外植体为带有１—２ ｍ ｍ 子叶柄的完整子叶,根癌农杆菌为Ａ２０８ＳＥ（ｐＴｉＴ３７－ＳＥ, ｐＲＯＡ９３）。Ｔｉ质粒ｐＲＯＡ９３ 带有ＮＰＴⅡ及ＧＵＳ嵌合基因。共培养２ 天后转到附加２５ ｍ ｇ／Ｌ卡那霉素的分化培养基（ＭＳ＋ ４．５ ｍ ｇ／ＬＢＡＰ）上。ＡｇＮＯ３ 和羧苄青霉素促进芽的分化,头孢霉素则有抑制作用。最高转化频率为２７％ 。把分化出的茎芽切下,插入含有２５ ｍ ｇ／Ｌ卡那霉素的生根培养基中。羧苄青霉素不利于根的形成。把完整抗性植株移入盛土壤的盆中,生长状况良好。测定β－葡糖苷酸酶活性,８４％ 明显高于对照。以ＮＰＴⅡ基因作探针进行Ｓｏｕｔｈｅｒｎ ｂｌｏｔ分析,证实外源基因已插入到植物细胞基因组中     

利用微束激光穿刺法将菜豆几丁质酶基因导入油菜的研究

利用微束激光穿刺法将菜豆几丁质酶基因导入油菜的研究杨仲毅王兰岚刘志岩张铁汉刘桂珍陈正华（中国科学院遗传研究所,北京,１００１０１）ＩｎｔｒｏｄｕｃｔｉｏｎｏｆＢｅａｎＣｈｉｔｉｎａｓＧｅｎｅｉｎｔｏＢｒａｓｓｉｃａｎａｐｕｓｂｙＬａｓｅｒＭｉｃｒｏｂ...     

甘蓝型油菜和芥菜型油菜种间杂交研究

甘蓝型油菜与芥菜型油菜杂交研究结果表明,杂交结实力与杂交组合方式以及参与杂交的亲本材料有关,以芥菜型油菜作母本的杂交结实力高于以甘蓝型作母本的组合：芥×甘杂交组合的平均结实数／花为 ２．６４ 粒,而甘×芥杂交组合的平均结实数／花为 ０．１０ 粒。芥甘杂种一代形态特征和生育期介于双亲之间,甘芥杂种一代不表现整齐的中间类型,株间差异明显;总体来看,芥甘杂种一代与双亲回交的结实力（０．４０,０．２１）低于甘芥杂种一代与双亲回交的结实力（３．３０,１．７４）,无论是芥甘杂种一代还是甘芥杂种一代,用甘蓝型油菜作父本回交的结实力高于用芥菜型油菜作父本回交的结实力,但也有个别回交组合出现例外,不表现上述规律。 Ｂ Ｃ１ 代种子当年播种出苗率低（１８．５％ ）,群体株间性状差异明显,生育期极不一致。芥甘杂种一代与甘蓝型油菜亲本第二次回交,其平均结实数／花较回交一代提高 １．０８ 粒, Ｂ Ｃ２ 代种子当年播种出苗率仍较低,但较对应的 Ｂ Ｃ１ 代稍有提高,群体中出现趋回交父本性状但雄性育性彻底退化的植株。芥甘杂种一代自由授粉所得 Ｆ２ 群体是一个变异极为丰富的遗传群体。     

Moricandia与芸苔属植物亲缘关系的研究

Ｍｏｒｉｃａｎｄｉａ是十字花科中唯一的具Ｃ３Ｃ４植物的属。为向十字花科芸苔属（Ｂｒａｓｉｃａ）作物中转移Ｃ３Ｃ４等重要性状提供基本信息,从可交配性和ＲＦＬＰ指纹图谱两个角度研究了Ｍｏｒｉｃａｎｄｉａ与芸苔属作物间的亲缘关系。结果表明：Ｍｏｒｉｃａｎｄｉａ与芸苔属作物间的可交配性很低,仅从Ｍ．ａｒｖｅｎｓｉｓ×甘蓝型油菜的１０５枚培养子房中获得４个杂种苗。用２３个甘蓝型油菜核ＤＮＡ探针和４个甜菜线粒体ＤＮＡ探针检测了芸苔属作物和ＭｏｒｉｃａｎｄｉａＣ３Ｃ４植物间ＤＮＡ限制性内切酶位点多态性。Ｍｏｒｉｃａｎｄｉａ与芸苔属物种间的同源性很高,其中白菜型油菜与Ｍ．ｎｉｔｅｎｓ之间在核基因组上的相似性甚至大于它与同属的甘蓝型油菜间的相似性。ＭｏｒｉｃａｎｄｉａＣ３Ｃ４物种与芸苔属作物在核ＤＮＡ和细胞质ＤＮＡ间的高度同源性,揭示了通过有性杂交或原生质体融合及染色体间重组的途径向栽培种转移重要基因的可能性。     

云南血竭的化学成分及抗真菌活性

云南血竭为剑叶龙血树（Ｄｒａｃａｅｎａ ｃｏｃｈｉｎｃｈｉｎｅｎｓｉｓ（Ｌｏｕｒ．）Ｓ．ｃ．Ｃｈｅｎ）树脂,从中分离到５个芳香族化合物：对羟基苯甲酸乙酰（１）,７,４＾′－二羟基黄烷（２）,７－羟基－４＾′－甲氧基黄烷（３）,７,４＾′－二羟基黄酮（４）和ｌｏｕｒｅｉｒｉｎ Ａ（５）以及１个甾体皂甙（６）,并对其中３个酚性成分进行了抗真菌活性检测。另外,用薄层层析法对云南血竭、广西血竭、海南血竭及     

鞭打绣球中的苯丙素甙和环烯醚萜甙

从鞭打绣球（ＨｅｍｉｐｈｒａｇｍａｈｅｔｅｒｏｐｈｙｌｌｕｍＷａｌｌ．）（玄参科）的全草中分离到２个新的苯丙素甙,命名为鞭打绣球甙Ａ和Ｂ（ｈｅｍｉｐｈｒｏｓｉｄｅＡａｎｄＢ）,２个已知的苯丙素甙,ｐｌａｎｔａｍａｊｏｓｉｄｅ和ｐｌａｎｔａｉｎｏｓｉｄｅＤ,以及３个已知的环烯醚萜甙,ｇｌｏｂｕｌａｒｉｃｉｓｉｎ,ｇｌｏｂｕｌａｒｉｎ和ｉｓｏ－ｓｃｒｏｐｈｕｌａｒｉｏｓｉｄｅ．通过化学和光谱分析,鞭打绣球甙Ａ和Ｂ的结构分别鉴定为２－（３－羟基－４－甲氧基苯基）乙基０－β－Ｄ－葡萄吡喃糖基（１→３）－４－Ｏ－反式阿魏醚基－β－Ｄ－葡萄吡喃糖试和２－（３,４－二羟基苯基）乙基Ｏ－［６－Ｏ－乙醚基－β－Ｄ－葡萄吡喃糖基（１→３）］－４－Ｏ－反式咖啡醚基－β－Ｄ－葡萄吡喃糖甙．     

两个油菜种对水分胁迫的适应方式

本文报道了2个油菜种对水分胁迫的适应方式。研究表明:芥菜型油菜对水分胁迫的适应性强于甘蓝型。其主要原因是由于形态方面的抗旱性,包括发育良好的根系,较厚的蜡质关闭了气孔。同时芥菜型油菜能将其余部分的水分调用到生长区而免遭旱害。甘蓝型油菜叶水势下降快,在相同水势下其相对电导值低于芥菜型。同时还观察了2个种在干旱条件下叶绿体与其基粒的变化。总之,芥菜型具典型的高水势耐旱特性,而甘蓝型具低水势耐旱特性。     

Variation of glucosinolates in vegetable crops of Brassica rapa

Glucosinolate levels in leaves were determined in a collection of 113 varieties of turnip greens (Brassica rapa L.) from northwestern Spain grown at two sites. Sensorial attributes were also assessed by a consumer panel. The objectives were to determine the diversity among varieties in total glucosinolate content and glucosinolate profile and to evaluate their sensory attributes in relation to glucosinolate content for breeding purposes. Sixteen glucosinolates were identified, being the aliphatic glucosinolates, gluconapin and glucobrassicanapin the most abundant. Other aliphatic glucosinolates, such as progoitrin, glucoalyssin, and gluconapoleiferin were relatively abundant in varieties with a different glucosinolate profile. Indolic and aromatic glucosinolate concentrations were low and showed few differences among varieties. Differences in total glucosinolate content, glucosinolate profile and bitterness were found among varieties, with a total glucosinolate content ranging from 11.8 to 74.0micromolg(-1) dw at one site and from 7.5 to 56.9micromolg(-1) dw at the other site. Sensory analysis comparing bitterness with variation in glucosinolate, gluconapin and glucobrassicanapin concentrations suggested that these compounds and their breakdown products are not the only determinants of the characteristic flavour of this vegetable. Other phytochemicals are probably involved on the characteristic bitter flavour. The varieties MBG-BRS0132, MBG-BRS0082, MBG-BRS0173, and MBG-BRS0184 could be good candidates for future breeding programs since they had high total glucosinolate content and good agronomic performance. The presence of glucoraphanin in some varieties should be studied more extensively, because this aliphatic glucosinolate is the precursor of sulforaphane, a potent anti-cancer isothiocyanate.     

Reducing progoitrin and enriching glucoraphanin in Braasica napus seeds through silencing of the GSL-ALK gene family

The hydrolytic products of glucosinolates in brassica crops are bioactive compounds. Some glucosinolate derivatives such as oxazolidine-2-thione from progoitrin in brassica oilseed meal are toxic and detrimental to animals, but some isothiocyanates such as sulforaphane are potent anti-carcinogens that have preventive effects on several human cancers. In most B. rapa, B. napus and B. juncea vegetables and oilseeds, there is no or only trace amount of glucoraphanin that is the precursor to sulforaphane. In this paper, RNA interference (RNAi) of the GSL-ALK gene family was used to down-regulate the expression of GSL-ALK genes in B. napus. The detrimental glucosinolate progoitrin was reduced by 65 %, and the beneficial glucosinolate glucoraphanin was increased to a relatively high concentration (42.6 μmol g(-1) seed) in seeds of B. napus transgenic plants through silencing of the GSL-ALK gene family. Therefore, there is potential application of the new germplasm with reduced detrimental glucosinolates and increased beneficial glucosinolates for producing improved brassica vegetables.     

Quantification of glucosinolates in leaves of leaf rape (Brassica napus ssp. pabularia) by near-infrared spectroscopy

The potential of near-infrared spectroscopy (NIRS) for screening the total glucosinolate (t-GSL) content, and also, the aliphatic glucosinolates gluconapin (GNA), glucobrassicanapin (GBN), progoitrin (PRO), glucoalyssin (GAL), and the indole glucosinolate glucobrassicin (GBS) in the leaf rape (Brassica napus L. ssp. pabularia DC), was assessed. This crop is grown for edible leaves for both fodder and human consumption. In Galicia (northwestern Spain) it is highly appreciated for human nutrition and have the common name of "nabicol". A collection of 36 local populations of nabicol was analysed by NIRS for glucosinolate composition. The reference values for glucosinolates, as they were obtained by high performance liquid chromatography on the leaf samples, were regressed against different spectral transformations by modified partial least-squares (MPLS) regression. The coefficients of determination in cross-validation (r2) shown by the equations for t-GSL, GNA, GBN, PRO, GAL and GBS were, respectively, 0.88, 0.73, 0.81, 0.78, 0.37 and 0.41. The standard deviation to standard error of cross-validation ratio, were for these constituents, as follows: t-GSL, 2.96; GNA, 1.94; GBN, 2.31; PRO, 2.11; GAL, 1.27, and GBS, 1.29. These results show that the equations developed for total glucosinolates, as well as those for gluconapin, glucobrassicanapin and progoitrin, can be used for screening these compounds in the leaves of this species. In addition, the glucoalyssin and glucobrassicin equations obtained, can be used to identify those samples with low and high contents. From the study of the MPLS loadings of the first three terms of the different equations, it can be concluded that some major cell components as protein and cellulose, highly participated in modelling the equations for glucosinolates.     

诸葛菜与芸苔属属间杂交时花粉—雌蕊相互作用的研究

本文对诸葛菜和芸苔属属间杂交的花粉粒萌发和花粉管生长情况进行了研究。其中用诸葛菜作父本,分别和甘蓝型油菜（“文油６号”、“云油３１号”）、“羽衣甘蓝”和白菜型油菜（“凤仪油菜”）杂交时,受精前有一定的亲和性;而和白菜型油菜（“玉溪周永达油菜”）、芥菜型油菜（“泸州四棱”、“南充油菜”）、埃塞俄比亚芥杂交时,存在受精前障碍。同时还发现通过蕾期授粉是克服属间杂交不亲和性的有效方法之一。     

Glucosinolate profiling of Brassica rapa cultivars after infection by Leptosphaeria maculans and Fusarium oxysporum

The glucosinolate contents of two different cultivars of Brassica rapa (Herfstraap and Oleifera) infected with Leptosphaeria maculans and Fusarium oxysporum were determined. Infection triggered the accumulation of aliphatic glucosinolates (gluconapin, progoitrin, glucobrassicanapin and gluconapoleiferin) and indole glucosinolate (4-hydroxy-glucobrassicin) in Herfstraap and of two indole glucosinolates (glucobrassicin and 4-hydroxy-glucobrassicin) in Oleifera. While total and aliphatic glucosinolates decreased significantly in Oleifera, a large increase was observed in Herfstraap after fungal infection. The indole glucosinolate glucobrassicin accumulated in Oleifera at a higher rate than Herfstraap especially after infection with F. oxysporum. Apparently the interaction between fungus and B. rapa is cultivar and fungal species specific.     

萝卜细胞质甘蓝型油菜“三系”研究初报

1984年3月,我们从蔬菜a血缘的材料中,找到了萝·甘油A[注]的恢复源;又经同年8月,在昆明夏季繁殖时,筛选出5个全恢复株系。这为萝·甘油A实现“三系”(不育系、保持系、恢复系)配套奠定了基础。这一最先发现,使国内外尚未解决萝·甘油A的恢复源和其杂种缺乏叶绿素问题得到了解决。     

远缘杂交油菜核不育系的创建及其细胞学和形态学研究

在甘蓝型油菜与诸葛菜以及芥菜型油菜与诸葛菜属间杂交后代中分别发现1个和3个不育材料,经杂交和多代近交育成了相应的甘蓝型油菜不育系。通过对核不育系体细胞鉴定表明,所有新发现的不育系染色体数为38,均已恢复到甘蓝型油菜。这些不育系绝大部分花粉母细胞（PMC）在中期Ⅰ、后期Ⅰ和后期Ⅱ 3个时期染色体行为表现正常,但不同时期的PMC均会出现一定比例的异常现象,主要表现为染色体落后或染色体桥等。这些不育系属于单核败育型,不育株与可育株的花器形态差异明显,不育系还存在不同程度的死蕾等特点。通过对花器生长过程的研究,发现不育株雌蕊生长随雄蕊败育进程逐渐加快,而可育株雌蕊生长则存在两个生长缓慢阶段。此外,文章还讨论了这些不育系的应用前景。     

油菜硫甙性状QTL 区间的加密和整合

目的:加密油菜控制硫甙性状QTL区间,并进行QTL整合预测候选基因。方法:利用生物信息学方法根据已知测序的白菜BAC序列信息设计引物,在油菜TN DH群体中进行多态性扩增和定位,并根据加密后构建的遗传连锁图重新检测QTL,进行QTL整合。结果:将根据白菜BAC设计的3对多态性标记成功定位到油菜控制硫甙性状QTL区间,进行QTL整合后将QTL置信区间进一步缩小,并判定了初步的候选基因。结论:充分利用白菜已测序的BAC或者基因组信息,将能加快油菜基础研究的步伐。     

芥蓝与芸苔属几个常见种花粉形态的比较

本文通过对芸苔属(Brassica L.)几个常见种芥蓝(B. alboglabra Bailey)、芥菜(B.juncea(L. )Coss.)、油菜(B. campestris L.)和花椰菜(B. oleracea var. botrytis L.)等花粉形态的光学显微镜和扫描电镜的观察比较,得知它们的花粉形态特征比较一致。而与杨萍等关于芥蓝的花粉形态特征——无极,具散孔这一报道截然不同;亦与黄增泉(Huang Tseng-chieng)关于芥菜的花粉特征——不具萌发孔的报道不同。作者的研究表明,上述几个种的花粉均具3沟,沟长而明显。外壁具网状纹饰,网眼较粗。但种与种间其花粉大小有所不同。作者不同意杨萍等关于“由于芥蓝的花粉形态与芸苔属其它种的花粉明显区别,说明芥蓝与芸苔属其它种不很亲缘”的观点,而认为,从花粉形态看,芥蓝与芸苔属其它种是基本一致的,把它们置于同一自然类群是合理的。