关于"烟草花叶病毒的感染和重建实验"的几个疑惑点探析

烟草花叶病毒的感染和重建实验是证明核酸是遗传物质的证据链中关键的一环.主要以科学家最初发表的2篇相关研究论文为依据,对学生在学习烟草花叶病毒的感染和重建实验过程中,经常产生的如何进行烟草花叶病毒感染烟草的操作、如何将烟草花叶病毒的RNA与蛋白质相互分离、如何实现烟草花叶病毒的体外重建3个疑惑点进行探析.     

烟草与紫苏科间远缘杂种后代化学成分及医药成分的研究

为利用药用植物资源以缓解吸烟与健康的矛盾,通过创新育种的方法将普通烟草品种78-04与紫苏进行科间远缘杂交,对选育出的新型烟草稳定品系—紫苏烟的内在化学成分及医药成分进行分析。酯酶与过氧化物酶同工酶分析证实,通过药用植物与烟草基因间的渐渗、交流以及重组,紫苏烟中转入了父本紫苏的遗传物质。烟叶主要化学成分分析显示,紫苏烟具有低糖、中高烟碱的特点,芸香苷的含量比亲本普通烟草品种78-04提高了70.4%。气相色谱-质谱联用技术初步分析表明,紫苏烟中含有α-丁香烯、β-丁香烯、芳樟醇和薄荷醇等紫苏挥发油的成分,与紫苏比较其薄荷醇的含量提高了2倍多;并产生了α-桉醇、香叶烯、桉油精、愈创木奥、α-郁金烯和γ-榄香烯等亲本所没有的医药成分和香料成分。药用植物与普通烟草科间远缘杂交能创造变异、产生烟草新类型,对降低卷烟产品的危害性具有一定作用,紫苏烟在医药和食品等领域也具有很高的综合利用价值。     

烟草与白花曼陀罗属间远缘杂种后代化学成分的研究

为缓解吸烟与健康的矛盾,通过常规育种的方法将普通烟草品种78-04与药用植物白花曼陀罗(Daturametel L.)进行属间远缘杂交育种,对选育出的新型烟草品系长柄曼陀罗烟(Mantuoluo tobacco)的内在化学成分和药用成分进行分析.酯酶同工酶和RAPD分析证实,长柄曼陀罗烟中转入了亲本药用植物的遗传物质.烟叶内在化学成分及药用成分分析显示,长柄蔓陀罗烟具有低糖、中高烟碱,含有东莨菪碱和阿托品等药用成分的特点.普通烟草与白花曼陀罗属间远缘杂交能创造变异、产生烟草新类型,对降低卷烟产品的危害性具有一定作用.     

三角鲂（Megalobrama terminalis）精子与青鱼（Mylopharyngodon piceus）卵子的受精细胞学研究

以青鱼(Mylopharyngodon piccus)为母本和三角鲂(Megalobrama terminalis)为父本的杂交虽是不同亚科的远缘杂交,但有正常的受精细胞学程序和常规的细胞分裂(卵裂)方式。这些实验结果,为开展鱼类远缘杂交提供了受精生物学的理论基础;同时还证实鱼类远缘杂交的异种精子不仅有激活卵子的作用,而且参与了遗传物质的组成,使父本的性状能够在杂种后代表现出来。实践证明,鱼类遗传育种可以通过远缘杂交的途径获得杂种优势。青鲂杂种一代既具有母本青鱼的性状,也兼有父本三角鲂的特征,通过养殖试验已在渔业生产中取得成效(拟另文发表)。     

低能离子束介导的遗传转化研究进展

低能离子束注入对细胞的刻蚀作用提供了外源遗传物质进入细胞的途径,离子束介导的遗传转化技术已在水稻,小麦,烟草,棉花等多种植物上取得成功,对离子束介导遗传转化的原理和最新进展进行了评述。．     

线粒体中的遗传物质

本文介绍了与高中生物课有关的线粒体遗传物质的复制、转录、蛋白质的合成,以及线粒体遗传物质和细胞核遗传物质之间的相互关系,可供师生在“细胞的结构和功能”一节教学中参考。     

核酸都是遗传物质吗?

《遗传变异》一章是高中生物教学中的重点,而“遗传的物质基础”则是重点中的重点和难点。但教材中一些模糊的说法却会使学生乃至教师产生错误的理解。例如,前不久笔者从不同的练习册上看到两条相似的题目: 1、组成人噬菌体烟草花叶病毒中核酸的核苷酸分别有多少种? A、8,4,4 B、4,4,4 C、5,4,4 D、8,4,5 2、组成人噬菌体烟草花叶病毒中遗传物质的核苷酸分别有多少种? 供选答案相同。一些同学和老师认为这两条题目都应选A,因为教材(最     

人工雌核发育鲢的遗传多样性及异源遗传物质整入的RAPD分析

运用RAPD技术对连续二代人工雌核发育鲢的遗传多样性及异源遗传物质的整入进行了分析 ,结果表明 :一代雌核发育鲢 ,个体间遗传相似度为 0 94 5— 0 995 6 ,多样性指数为 0 175 ;二代雌核发育鲢 ,个体间遗传相似度为0 96 15— 1 0 0 ,平均为 0 985 2 ,多样性指数为 0 0 6 2。研究揭示经过连续二代人工雌核发育后 ,其遗传多样性明显减少 ,种质进一步纯化。通过对雌核发育鲢二代、亲本鲢和雄鲤的RAPD扩增比较 ,发现雌核发育鲢含有少数与父本相同的特异DNA扩增带 ,而亲本鲢没有 ,在基因水平上表明雌核发育鲢整入了雄鲤的遗传物质     

美科学家将外源基因导入叶绿体

美国纽约州华斯曼研究所的科学家通过基因工程,已将外源遗传物质导入植物的叶绿体,并使这种外源DNA稳定地整合于其后代的叶绿体。上述研究是基因工程上的突破。它可使科学家有可能操纵涉及光合作用的基因,阐明光合作用的某些最基本的过程,从而进一步改造植物和改良农作物品种。这些科学家以烟草N.tabacum SPC2为试材。这种烟草的质粒pZS148(9.6千碱基)含有可编码壮观霉素抗性突变的DNA片段。用这种DNA包衣的钨粒子轰击烟草植株的叶组织。然后通过叶组织培养的壮观霉素抗性     

异源精子遗传物质对雌核发育草鱼基因组的影响分析

为分析外源精子对人工诱导雌核发育草鱼基因组的影响,用随机扩增多态性和微卫星技术对经两代连续人工诱导,遗传背景一致的雌核发育草鱼以及母本草鱼和父本鲤鱼的基因组DNA进行了比较分析。10个随机引物在雌核发育草鱼中共检测到104个RAPD位点,在鲤鱼中检测到103个位点。7对微卫星引物在雌核发育草鱼中共扩增出4个微卫星位点,在鲤鱼中检测到22个位点。两种方法在雌核发育草鱼和鲤鱼中所检测到的位点均没有一个相同。根据RAPD和微卫星分析数据进行的遗传相似度分析表明二代人工雌核发育草鱼群体与其一代人工诱导雌核发育草鱼母本的遗传相似度从0.9903到1.000,与父本鲤鱼的遗传相似度为0.000。这些实验结果证明在适当的紫外线处理强度下,鲤鱼精子的遗传物质能够被完全破坏,不会对雌核发育草鱼的基因组造成遗传污染。     

Intergeneric transfer of cytoplasmic male sterility between Raphanus sativus (cms line) and Brassica napus through cytoplast-protoplast fusion

Summary Cytoplasts isolated from hypocotyl protoplasts of Raphanus sativus cv Kosena (cms line) by ultracentrifugation through Percoll/mannitol discontinuous gradient were fused with iodoacetamide(IOA)-treated protoplasts of Brassica napus cv Westar. Seventeen randomly selected regenerated plants were characterized for morphology and chromosome numbers. All of the regenerated plants had morphology identical to B. napus and 10 of them possessed the diploid chromosome number of B. napus. The remaining plants had chimeric or aneuploid chromosome numbers. The mitochondrial genomes in the 10 fusion products possessing the diploid chromosome numbers of B. napus were examined by Southern hybridization analysis. Four of the 10 plants contained mitochondrial DNA showing novel hybridization patterns. Of these 4 plants, 1 was male sterile, and 3 were male fertile. The remaining plants showed mitochondrial DNA patterns identical to B. napus and were male fertile.     

Morphological, cytogenetic, and molecular evidence for introgressive hybridization in birch.

Extensive morphological variation of tetraploid birch (Betula pubescens) in Iceland is believed to be due to gene flow from diploid dwarf birch (B. nana) by means of introgressive hybridization. A combined morphological and cytogenetic approach was used to investigate this phenomenon in two geographically separated populations of natural birch woodland in Iceland. The results not only confirmed introgressive hybridization in birch, but also revealed bidirectional gene flow between the two species via triploid interspecific hybrids. The populations showed continuous morphological variation connecting the species, but karyotypically they consisted of only three types of plants: diploids, triploids, and tetraploids. No aneuploids were found. Some of the tetraploid plants had B. pubescens morphology as expected, but most of them had intermediate characters. Most of the diploid plants were B. nana, but some were intermediates and a few had B. pubescens morphology. The triploid plants were either intermediates or they resembled one of the two species. Similar introgressive variation was observed among the diploid and triploid progeny of open-pollinated B. nana in a garden. Birch samples including field plants and artificial hybrids were further examined using a molecular method based on genomic Southern hybridization. The experiments verified introgression at the DNA level.     

Bioengineering of a phytoremediation plant by means of somatic hybridization

Phytoremediation is a technology that exploits a plant's ability to remove contaminants from the environment or render toxic compounds harmless. An efficient metal phytoremediating plant must combine high biomass production and established cultivation methods with high tolerance to a specific contaminant and ability for root uptake, translocation, and compartmentalization of contaminants in the above-ground biomass. Symmetric and asymmetric somatic hybridizations were used to introduce toxic metal-resistant traits from Thlaspi caerulescens into Brassica juncea. B. juncea hypocotyl protoplasts were fused with T. caerulescens mesophyll protoplasts. The hypocotyl protoplasts of B. juncea were stained with CFDA before fusion and thus fluoresced green under UV, whereas the mesophyll protoplasts of T. caerulescens had red autofluorescense. Heteroplasmic fusion products were identified and selected by flow cytometry and cell sorting. All putative hybrids grown in the greenhouse had morphological characteristics of B. juncea. A Thlaspi-specific repetitive sequence was hybridized to total DNA of plants, including the parental species. All plants from both symmetric and asymmetric fusions showed Thlaspi-specific hybridization patterns while B. juncea did not exhibit any hybridization signal. Hybrid plants, produced by asymmetric somatic hybridization between the two species, demonstrated high metal accumulation potential, tolerance to toxic metals, and good biomass production.     

芥菜型油菜和白菜型油菜种间杂种遗传分析

种间杂交是一种拓宽栽培作物遗传基础和转移优良性状的重要手段,已经广泛地用于作物品质的改良。本研究通过芥菜型油菜（Brassica juncea L.）和白菜型油菜（Brassica rapa L.）种间杂交,将芥菜型油菜的有利性状转移到白菜型油菜中,创造新型白菜型油菜,以改良白菜型油菜的农艺性状、提高抗逆性和拓宽其遗传基础。研究结果表明：以芥菜型油菜作母本、白菜型油菜作父本的杂交组合较易获得杂交种子,杂种F1植株营养生长具有较明显的杂种优势,但花粉完全不育;以白菜型油菜回交获得的BC1植株间表型差异明显,平均花粉可染率为34.8%,介于 0~84%之间,群体自交不亲和;BC1F1和BC2群体变异广泛,出现自交亲和植株和黄籽植株,平均花粉可染率分别为79.7%和79.1%。     

Production and analysis of asymmetric hybrid plants between monocotyledon (Oryza sativa L.) and dicotyledon (Daucus carota L.)

Asymmetric hybrid plants were obtained from fused protoplasts of a monocotyledon (Oryza sativa L.) and a dicotyledon (Daucus carota L.). X-ray-irradiated protoplasts isolated from a cytoplasmic malesterile (cms) carrot suspension culture were fused with iodoacetoamide-treated protoplasts isolated from a 5-methyltryptophan (5MT)-resistant rice suspension culture by electrofusion. The complementary recovered cells divided and formed colonies, which were then cultivated on regeneration medium supplemented with 25mg/l 5MT to eliminate any escaped carrot cells. Somatic hybrids were regenerated from 5 of the 5MT-resistant colonies. The morphologies of most of the regenerated plants closely resembled that of the parental carrot plants. A cytological analysis of callus cultures induced from these plants indicated that most of the cells possessed 20–22 chromosomes and were resistant to 5MT. An isozyme analysis revealed that several regenerated plants had the peroxidase isozyme patterns of both parents. A Southern hybridization analysis with non-radioactively labelled DNA fragments of the rgp1 gene showed that regenerated plants had hybridizing bands from both rice and carrot. Chloroplast (cp) and mitochondrial (mt) DNAs were also analyzed by Southern hybridization by using several probes. CpDNA patterns of the regenerated plants were indistinguishable from those of the carrot parent. However 1 of the regenerated plants had a novel band pattern of mtDNA that was not detected in either of the parents, indicating a possible recombination of mitochondrial genomes.     

Production of somatic hybrids between Daucus carota L. and Nicotiana tabacum

Protoplasts of a kanamycin-resistant (KR, nuclear genome), streptomycin-resistant (SR, chloroplast genome) and chlorophyll-deficient (A1, nuclear genome) Nicotiana tabacum (KR-SA) cell suspension cultures or X-ray-irradiated mesophyll protoplasts of kanamycin- and streptomycin-resistant green plants (KR-SR) were fused with protoplasts of a cytoplasmic male-sterile (CMS) Daucus carota L. cell suspension cultures by electrofusion. Somatic hybrid plants were selected for kanamycin resistance and the ability to produce chlorophyll. Most of the regenerated plants had a normal D. carota morphology. Callus induced from these plants possessed 23–32 chromosomes, a number lower than the combined chromosome number (66) of the parents, and were resistant to kanamycin, but they segregated for streptomycin resistance, which indicated that N. tabacum chloroplasts had been eliminated. Genomic DNA from several regenerated plants was analyzed by Southern hybridization for the presence of the neomycin phosphotransferase gene (NPTII); all of the plants analyzed were found to contain this gene. Mitochondrial (mt) DNA was analyzed by Southern hybridization of restriction endonuclease digests of mtDNA with two DNA probes, PKT5 and coxII. The results showed that the two plants analyzed possessed the mitochondria of D. carota. These results demonstrate that the regenerated plants are interfamilial somatic hybrids.     

Fusion of male-sterile tobacco causes modifications of mtDNA leading to changes in floral morphology and restoration of fertility in cybrid plants

Protoplasts from male-sterile Nicotiana tabacum cultivars with cytoplasms of N. suaveolens, N. bigelovii and N. undulata were fused in different pairwise combinations. Cybrid plants were obtained and categorized according to their flower morphologies into groups as parental male-sterile types, recombined biparentals, novel male-steriles and male fertiles. Restriction enzyme analyses and DNA gel blot hybridizations of the mitochondrial DNA (mtDNA) of these cybrid plants revealed that all plants with parental male-sterile morphologies had banding patterns identical to the parental patterns, whereas all cybrids with flower morphologies different from their parents had novel patterns. Several restriction fragments were found to be unique to the mtDNA of male-fertile plants with perfectly restored stamen morphologies when compared to the mtDNA from cybrid plants with incomplete restoration. MtDNA gel blot hybridization analysis of fertile plants revealed that the probe for atpA hybridized to two restriction fragments, one from each parent. Further, fertile plants with complete stamen restoration lacked an expressed sequence in the 3'flanking region of atpA , in contrast to both male-sterile parents.     

Hybridization of Neotyphodium endophytes enhances competitive ability of the host grass

? Associations with microbial symbionts may lead to niche differentiation of their host. Vertically transmitted Neotyphodium endophytes of grasses often hybridize in nature. Infection by these hybrid symbionts may result in different host-plant phenotypes from those caused as a result of infection by nonhybrid symbionts. Observations of wild Arizona fescue (Festuca arizonica) populations show that hybrid Neotyphodium-infected (H+) grasses dominate in resource-poor environments, whereas nonhybrid endophyte-infected (NH+) grasses dominate in environments with more resources. We studied the hypothesis that hybridization of endophytes increases stress tolerance of the host. ? To test whether hybridization of Neotyphodium affects performance and competitive abilities of the host depending on resources, we conducted a glasshouse experiment where competition, nutrients and watering were manipulated. ? H+ plants had greater wet biomass than NH+ and endophyte-free plants, when grown in competition, but only in low-water and low-nutrient treatments. By contrast, NH+ plants did not perform better than H+ or endophyte-free plants regardless of the treatment combination. ? Our results suggest that hybridization of symbiotic Neotyphodium endophytes may increase competitive potential of the host in stressful environments and that this hybridization may be underlying niche expansion of Arizona fescue in the environments with low resources.     

苜蓿红豆草属间体细胞杂种的分子生物学鉴定

通过原生质体融合和培养获得苜蓿红豆草属间体细胞杂种植株。采用一种简便方法从杂种组织再生植株叶片、红豆草羟脯氨酸抗性系再生植株叶片和苜蓿根癌农杆菌702转化系愈伤组织提取DNA用于RAPD和Southern杂交分析。随机引物扩增结果显示两种亲本的RAPD多态具有明显差异。在所用20种随机引物中,6种产生较多的DNA片段。杂种组织具有两种亲本特有的DNA片段,但倾向于排除红豆草亲本的染色体,表明该杂种为非对称杂种,两种亲本染色体之间可能发生了重组。由于红豆草DNA的介入,杂种组织表现出较强的分化能力。分别利用RAPD扩增得到的OPA141000bp红豆草羟脯氨酸抗性系特异产物和OPA141600bp苜蓿根癌农杆菌702转化系特异产物为探针进行Southern分子杂交,证明杂种组织同时具有这两种DNA片段的同源序列。     

Production and analysis of plants that are somatic hybrids of barley (Hordeum vulgare L.) and carrot (Daucus carota L.)

In order to obtain plants that were somatic hybrids of barley (Hordeum vulgare L.) and carrot (Daucus carota L.), we fused protoplasts that had been isolated from 6-month-old suspension cultures of carrot cells with protoplasts isolated from barley mesophyll by electrofusion. After culture for 1 month at 25°C , the cells were cultured for 5 weeks at 4°C , and were then returned to 25°C for culture on a shoot-inducing medium. Three plants (nos. 1, 2 and 3) were regenerated from the cells. The morphology of the regenerated plants closely resembled that of the parental carrot plants. A cytological analysis of callus cultures induced from these plants indicated that most of the cells had about 24 chromosomes, fewer than the sum of the numbers of parent chromosomes which was 32. Southern hybridization analysis with fragments of the rgp1 gene used as probe showed that the regenerated plants contained both barley and carrot genomic DNA. Chloroplast (ct) and mitochondrial (mt) DNAs were also analyzed with several probes. The ctDNA of the regenerated plants yielded hybridization bands specific for both barley and carrot when one fragment of rice ctDNA was used as probe. Furthermore, the regenerated plants yielded a barley specific band and a novel band with another fragment of rice ct DNA as a probe. One of the regenerated plants (no. 1) yielded a novel pattern of hybridized bands of mt DNA (with an atp6 probe) that was not detected with either of the parents. These results indicated that the regenerated plants were somatic hybrids of barley and carrot and that recombination of both the chloroplast genomes and the mitochondrial genomes might have occurred. Received: 28 May 1996 / Accepted: 2 August 1996