诸葛菜组织培养中的器官形成

植物组织和细胞培养技术已在农作物的改良及园艺植物的快速繁殖方面得到广泛的应用。在这方面,十字花科植物正在受到相当多的注意。新近,为了建立十字花科植物的细胞转化系统,我们试验了多种十字花科植物,发现诸葛菜的叶和叶柄等外植体具有极强的器官分化能力,现将结果报告如下。本文取材植物诸葛菜,又名二月兰(Oryc-hophragmus violaceus/Moricandia sonchifolia),     

Transformation of Orychophragmus violaceus Using Agrobacterium tumefaciens And Regeneration of Transgenic Plants

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甘蓝型油菜与诸葛菜属间杂种无性系的变异研究

从形态及细胞学两方面对甘蓝型油菜与诸葛菜属间杂种无性系的变异进行了研究。经过长期继代培养后,杂种在形态上越来越偏向母本甘蓝型油菜,而表现出较少的父本诸葛菜性状,但表现出对等双分枝的新性状。经细胞学观察表明,杂种体内杂种细胞比例下降,而甘蓝型油菜细胞比例大幅度上升并远高于核质杂种细胞（具有油菜细胞质与诸葛菜细胞核）的比例,以至较多的甘蓝型油菜染色体组被遗传下去。在该杂种继代培养中还观察到杂种细胞内的染色体消除及体细胞配对现象。     

Production and cytogenetics of intergeneric hybrids between Brassica napus and Orychophragmus violaceus

The intergeneric hybrid between Brassica napus and Orychophragmus violaceus was obtained by means of embryo culture technique with the latter as the pollen parent. The hybrid was morphologically intermediate between its parents, but could produce a lot of seeds when selfed. Somatic separation of the genomes from the two parental species was observed during the mitotic divisions of some of the hybrid cells. Thus, the hybrid became the mixoploid in nature, consisting of haploid and diploid cells of B. napus, and a nuclear — cytoplasmic hybrid, with the cytoplasm of B. napus and the nuclei of O. violaceus, and the hybrid cells. Pollen mother cells with 19, 12 and 6 bivalents, respectively, were produced by the hybrid. From the selfed progeny of the hybrid, mainly two kinds of plants, B. napus and the hybrid, were found. The hybrid plants of the selfed progeny again produced two kinds of plants, B. napus and the hybrid.     

转OvDREB2B基因陆地棉鉴定及其对水分渗透胁迫的分析

通过花粉管通道技术,以该实验室自育陆地棉品系TH₁和TH₂为材料,将诸葛菜(Orychophragmus vidaceus)抗逆转录因子OvDREB2B基因构建到植物表达载体后,导入棉花基因组,经卡那霉素筛选和分子鉴定表明目的基因已整合到棉花基因组中并表达。将T₁代转基因植株和受体对照在温室中栽培,待植株生长至四叶一心时,用不同渗透势的PEG-6000水溶液进行渗透胁迫处理,分析探讨转基因植株的抗旱效果及其抗旱机理。结果显示:当渗透势为0和0.5 MPa处理时,转基因植株和对照无明显差异;当渗透势为0.8 MPa和1.1 MPa处理时转基因植株较对照抗旱性明显提高。当渗透势为1.1 MPa处理96 h时,对照植株F_v/F_m降至0.2左右,而转基因植株仍正常生长,F_v/F_m值约为0.51,而且初始荧光（F₀）值、净光合速率（P_n）、胞间CO₂浓度（C_i）、蒸腾速率（T_r）等一系列参数转基因植株都明显优于对照,表明DREB2B基因能够提高棉花对水分胁迫的耐受性。     

Origin of new <Emphasis Type="Italic">Brassica</Emphasis> types from a single intergeneric hybrid between <Emphasis Type="Italic">B. rapa</Emphasis> and <Emphasis Type="Italic">Orychophragmus violaceus</Emphasis> by rapid chromosome evolution and introgression

Many novel lines were established from an intergeneric mixoploid between Brassica rapa (2n = 20) and Orychophragmus violaceus (2n = 24) through successive selections for fertility and viability. Pedigrees of individual F₂ plants were advanced to the 10th generation by selfing. Their breeding habit was self-compatible and different from the self-incompatibility of their female parent B. rapa, and these lines were reproductively isolated to different degrees from B. rapa and B. napus. The lines with high productivity showed not only a wide spectrum of phenotypes but also obvious variations in fatty acid profiles of seed oil and glucosinolate contents in seed meal. These lines had 2n = 36, 37, 38, 39 and 40, with 2n = 38 being most frequent (64.56%), and no intact O. violaceus chromosomes were detected by genomic in situ hybridization (GISH) analysis. Amplified fragment length polymorphism (AFLP) analyses revealed a high extent of variation in genomic compositions across all the lines. O. violaceus-specific bands, deleted bands in B. rapa and novel bands for two parents were detected in these lines, with novel bands being the most frequent. The morphological and genetic divergence of these novel types derived from a single hybrid is probably due to rapid chromosomal evolution and introgression, and provides new genetic resources for rapeseed breeding.     

缺锌和HCO_3~-处理对诸葛菜和油菜有机酸特征的影响

以诸葛菜和油菜为材料,水培环境下设置4个不同的缺锌和碳酸氢根离子胁迫处理,分别为+Zn0(含Zn且不加HCO3-的处理组),+Zn10(含Zn且加10 mmol·L-1HCO3-的处理组),-Zn0(缺Zn且不加HCO3-的处理组)和-Zn10(缺Zn且加10 mmol·L-1HCO3-的处理组),利用离子色谱法分析了4个处理的两种植物幼苗器官(根、茎、叶)及根系分泌物中的有机酸特征。结果表明:(1)高浓度碳酸氢根离子处理显著增加了两种植物器官及根系分泌的有机酸总量,尤其是在缺锌和高浓度碳酸氢根离子双重胁迫下(-Zn10处理),诸葛菜器官和根系分泌的有机酸比油菜更敏感,草酸、柠檬酸和苹果酸是诸葛菜器官和根系分泌物中的优势酸,这三种有机酸的含量分别占其有机酸总量的75%及以上;(2)叶片是两种植物有机酸产生的主要器官,有机酸的含量和分配比例从地上部分(叶和茎)到地下部分(根)减少;(3)两种植物器官和根系分泌物中的有机酸变化趋势一致,叶片中有机酸主要来源于暗呼吸过程和光呼吸过程,其他器官和根系分泌物中的有机酸主要来源于暗呼吸过程;(4)诸葛菜对缺锌和高浓度碳酸氢根离子的适应能力强于油菜,为诸葛菜的喀斯特适生性和低锌和高浓度碳酸氢根离子环境(如喀斯特环境)的生态修复提供了理论依据。     

二月兰形态性状的变异分析

二月兰(Orychophragmus violaceus)是集食用、保健、饲用和观赏于一身的优良地被植物。其遗传背景单一、生长周期短,是发展模式植物的优良材料。为了开发其育种价值,我们对二月兰的形态变异式样进行了初步分析。共选取了北京和南京地区的7个居群的34个表型性状进行观测。通过计算分析发现:株高、茎和花序分枝等性状受环境影响较大,因而其分类价值不大;叶形、毛被、叶缘锯齿等性状的变异在居群内是连续的,所以也不是可靠的分类性状;而花部性状的一些变异,包括花瓣的颜色、大小、形态,基本不受环境影响,且这些变异在居群内是不连续的,所以其分类价值相对较高。对17个数值性状的方差分析和多重比较结果表明,在同一年度内,大部分性状在不同地区间差异不显著,在居群间差异极显著;而在不同年度内,居群间差异不显著。最后,对7个居群和30个表型性状进行了数量分类:Q分析表明北京颐和园、南京紫金山山顶公园和南京中山植物园居群比较相似,而北京圆明园、北京海淀区后八家苗圃、南京情侣园和北京天坛公园居群较相似。R分析揭示了二月兰花部、叶部、反映植株毛被和颜色以及表现植株生长量的性状间均具有一定的相关性。这些研究可为种下等级的划分和进一步的遗传研究以及品种培育提供一定的参数资料。     

埃塞俄比亚芥与诸葛菜属间杂种的基因组原位杂交分析

用幼胚培养方法重新获得的埃塞俄比亚芥(Brassica carinata A.Braun,2n=34)与诸葛菜(Orychophragmus violaceus (L.)O.E.Schulz,2n=24)的属间杂种仍为混倍体(2n=14～34),2n=34细胞的频率最高;绝大多数花粉母细胞(PMCs)表现正常的17个二价体配对和17：17的分离。基因组原位杂交分析结果表明,在所有体细胞和PMCs中不含有整条的诸葛菜染色体,2n=34的体细胞和PMCs中包含了来自黑芥(B．nigra (L．)Koch,2n=16)的16条染色体。这些具有完全或部分埃塞俄比亚芥染色体组成的细胞,可能来源于以前提出的杂种细胞在有丝分裂中完全或部分亲本染色体组分开和染色体复制,并伴随诸葛菜染色体的消除。     

Nuclear reconstitution of demembranated Orychophragmus violaceus sperm in Xenopus laevis egg extracts

The cell-free extracts from animal Xenopus laevis egg could induce chromatin decon-densation and pronuclear formation from demembranated plant (Orychophragmus violaceus) sperm. The demembranated Orychophragmus violaceus sperm began to swell in 30 min incubation, and then were gradually decondensed. The reassembly of nuclear envelope in the reconstituted nuclei had been visualized by means of electron microscopy and fluorescent microscopy. Membrane vesicles fused to form the double envelope around the periphery of the decondensed chromatin. The morphology of the newly formed nucleus, with a double membrane, was similar to those nuclei after fertilization. Transmission electron microscope micrograph of the whole mount prepared nuclear matrix-lamina showed the reconstituted nucleus to be filled with a dense network.