A_2型高粱细胞质雄性不育系与其保持系的胞质DNA和核DNA差异

以高粱细胞质雄性不育系A2 V4 (A)及其保持系V4 (B)的总DNA为模板 ,对 184个随机引物进行筛选 ,找到6个其RAPD扩增产物在A/B间存在稳定差异的引物 .将该 6个引物同时扩增A/B的总DNA、线粒体DNA(mtDNA)及叶绿体DNA(cpDNA) .以总DNA为模板时得到 12个扩增片段 ,以mtDNA为模板时得到 4个 ,以cpDNA为模板时得到 11个 .结果分析表明 ,在这些扩增片段中 ,有 7个仅仅出现在以胞质DNA为模板的扩增中 ,有 5个在以总DNA和胞质DNA为模板时同时出现 ,即认为这 12个片段来自胞质DNA .另有 7个片段 ,在以胞质DNA为模板时未出现 ,而是仅仅出现在以总DNA为模板的扩增中 ,认为是来自核DNA .来自核DNA的 7个扩增片段中 ,有 5个来自保持系 ,有 2个来自不育系 ,这表明 ,不育系与保持系在核DNA上存在差异 .对A/B核DNA在CMS中的重要性及研究对策进行了讨论 .图 2表 4参 19     

A2型高粱细胞质雄性不育系与其保持系的胞质DNA和核DNA差异

以高粱细胞质雄性不育系A2V4（A）及其保持系V4（B）的总DNA为模板，对184个随机引物进行筛选，找到6个其RAPD扩增产物在A／B间存在稳定差异的引物，将该6个引物同时扩增A／B的总DNA、线粒线DNA（mtDNA）及叶绿体DNA（cpDNA），以总DNA为模板时得到12个扩增片段，以mtDNA为模板时得到4个，以cpDNA为板时得到11个，结果分析表明，在这些扩增片段中，有7个仅仅出现在以胞质DNA为模板的扩增中，有5个在以总NDA和胞质DNA为模板时同时出现，即认为这12个片段来自胞质DNA，另有7个片段，在以胞质DNA为模板时未出现，而是仅仅出现在以总DNA为模板的扩增中，认为是来自核DNA，来自核DNA的7个扩增片段中，有5个来自保持系，有2个来自不育系，这表明，不育系与保持系在核DNA上存在差异，对A／B核DNA在CMS中的重要性及研究对策进行了讨论。     

CMS高粱保持系叶绿体基因ps1A1和ps1A2片段的克隆与序列比对

以15种包括同质异核和同核异质高粱材料的总：DNA为模板进行RAPD分析，196个随机引物中，引物Y-19扩增得到一个很有规律的差异片段SAY-192300。该片段只出现在4个保持系(B)和3个恢复系(R)中，而不出现在6个不育系(A)和2个杂种F1中，进一步对cms—Al、cms—A2两套8个材料(A／B／R／F1)的总DNA、mtDNA和cpDNA用引物Y-19进行扩增，发现片段SAY-192300仅在cpDNA得到，Southern杂交结果证实了片段SAY-192300的叶绿体来源，同时也表明在cms胞质中该片段所在区未发生插入、缺失等大的结构变异,DNA序列测定结果表明，该片段为叶绿体ps1A1和ps1A2基因的部分序列。图6表2参19。     

植物线粒体基因转录水平表达研究方法的建立

以茎瘤芥雄性不育系为材料,在已有文献的基础上,对常规方法进行改进,使植物线粒体分离、线粒体切片观察、线粒体RNA提取、反转录方法、PCR扩增、northern杂交等方法系统化,建立了一整套比较有效的适合于植物线粒体基因转录水平表达分析的方法.图4参12     

植物非编码RNA与雄性不育

利用雄性不育系做母本生产杂交一代种子,为改善作物的生产提供了十分有利的条件.细胞质雄性不育(CMS)和由光周期(PGMS)/温度诱导(TGMS)的不育性极大地促进了作物杂交育种的发展.越来越多的研究表明非编码ncRNA分子与植物雄性不育有关,如水稻的PGMS和TGMS与ncRNA密切相关.在CMS中,线粒体ORF影响小孢子发育而导致CMS发生的机制仍然不明确.高通量测序使得在全基因组范围内发现和验证这些调控分子及其靶基因,并阐述它们与CMS的相关性成为可能.植物线粒体DNA中的ncRNA以及其可能诱导的CMS的机理是当前研究的热点,但目前尚没有证据表明ncRNA与CMS有关,还需要转基因等遗传方法来进行验证.在本文中,我们总结了最近关于ncRNA与植物CMS关系的相关研究,希望能够提高对控制植物CMS的ncRNA介导的调控途径的理解.深入了解植物CMS可为促进CMS在农作物育种的利用提供技术支撑.     

玉米多胞质系线粒体数量和结构特征的比较分析

以玉米多胞质系为材料,采用透射电镜的观察方法,比较观察了１１种材料中黄化幼芽细胞的线粒体结构特征的变化．观察表明,１１种材料黄化幼芽线粒体形态及外膜、内膜、泡状嵴和基质颗粒等没有什么显著的差异,而线粒体大小和每个细胞内的线粒体数量却是明显不同的．Ｆ检验说明,线粒体大小和每个细胞线粒体数量上有极显著的差异,也即是说,１１种不同细胞质类型的材料细胞质作用和核质（基因）互作是明显的．试验还观察到线粒体、分裂现象和线粒体、淀粉粒聚集在胞间连丝附近的有趣现象．     

编后

本期所发《Ａ2 型高粱细胞质雄性不育系与其保持系的胞质ＤＮＡ和核ＤＮＡ差异》一文在审稿专家和编委中存在较大争议 ,其审处过程可说是“一波三折” ,“三起三落” .为此 ,我部召集部分在蓉编委听取了作者的申辩 ,虽然分歧和争议仍然存在 ,但最后决定予以刊出。发表此文的理由除保护作者科学民主权利外 ,还希望引起更多的人对此文所论问题的关注 .国际编辑界有一句用以协调审稿分歧意见的名言 :“如果审稿人有激烈的分歧 ,就干脆发表这份稿子 ;内中一定有某些令人兴奋的东西”(引自[英 ]Ｍ .奥康诺尔 :科技书刊编辑工作 .北京 :人民教育出…     

白菜叶绿体DNA快速提取及分析

采用SDS-蛋白酶法提取了白菜的叶绿体DNA.琼脂糖电泳及紫外分光光度分析表明,所提取的叶绿体DNA质量高,除cpDNA主环外,在胞质雄性不育新种质及原不育系中还存在约1375bp和831bp的两条质粒分子.用所提取的叶绿体DNA作为模板进行RAPD扩增,获得了清晰的扩增图谱.该方法与传统的提取方法相比省时、高效、无污染.图2表1参14     

两栖动物线粒体基因组tRNA基因重复、丢失及其影响

在脊椎动物线粒体基因组中,偶有tRNA基因重复或丢失事件发生,这些事件因与相应氨基酸转运密切关联而凸显重要.两栖类中,这些重复或丢失相对多样,可为相关研究提供良好素材.为了解这些事件对氨基酸对应的密码子使用偏好是否产生影响,本文引入统计方法,检测10种两栖类mtDNA蛋白编码基因相对的同义密码子使用度(RSCU),并特别设计将发生tRNA基因重复或丢失物种与其近缘群进行对比分析.结果显示,tRNA基因的重复对密码子使用偏好没有显著的影响,推测可能与突变积累的时限和mtDNA紧凑性有关.另外,tRNA基因丢失也未使相应氨基酸密码子使用偏好产生明显的改变.推测在两栖类中,"tRNA转入"和"tRNA反密码子摆动位点的超级摆动"可能是tRNA基因丢失的补偿方式.     

籼爪交后代的花药培养效率

对爪畦稻J15、J21、J20以及J211与不同的籼型杂交稻三系亲本杂交后代不同世代进行了花药培养.结果表明:当同一爪哇稻与三系亲本杂交时,其F1花药培养力高低顺序为:爪哇稻/保持系>爪哇稻/恢复系>爪哇稻/不育系的杂种后代.这种差异可能存在细胞质效应.4个爪哇稻品种中,以J15所配组合的出愈率和培养力为最高,绿苗分化率以J20所配组合最高.低代材料花药在适合籼粳交的SK3培养基上反应较好,高代材料的花药在适合籼稻的M8培养基上反应较好.改良M8和改良SK3培养基的花培效果分别优于M8和SK3培养基.F5、F6代材料的花药在改良M8培养基上出愈率可超过50％.琼脂糖代替琼脂可显著提高出愈率和培养力.高温预培养(30-32℃)24 h其花药培养效率达到最高值.图2表3参15     

核基因组在细胞质雄性不育中的作用研究：背景与现状

细胞质雄性不育 (ｃｙｔｏｐｌａｓｍｉｃｍａｌｅｓｔｅｒｉｌｉｔｙ ,ＣＭＳ) ,表现为小孢子发育异常、不能产生花粉或花粉败育 ,但卵细胞发育正常、可接受外来花粉而正常结实 ,系母性遗传 ,即细胞质遗传[1～ 4 ] .ＣＭＳ的重要性 ,首先表现为在农业生产上的巨大经济应用价值[2～ 4 ] .杂种优势的发现和利用为粮食生产带来了革命 ,但杂种种子的生产过程却是很繁杂的 ,核心是要阻止母本的自花授粉结实、保证母本植株上所结的种子全部是杂种 ,即种子的纯度很高才能保证农业生产上的应用 .ＣＭＳ ,由于其不能产生可育的花粉 ,即无自花授粉结…     

Mitochondrial DNA analysis of native and selectively inbred Chesapeake Bay oysters,Crassostrea virginica

Mitochondrial DNA (mtDNA) genotypes were examined in 1989 in threeCrassostrea virginica (Gmelin) populations native to Chesapeake Bay, USA, and in one population selectively inbred for rapid growth for ten generations. We wished to determine whether this character would be useful as a genetic marker for distinguishing between the inbred line and the native oysters and to determine whether detectable genetic differences exist among present-day native populations ofC. virginica. Thirty mtDNA haplotypes were identified. The average percentage nucleotide difference between native haplotypes was 1.8%. Inbred oysters were characterized by mtDNA haplotypes distinctly different from ancestral native oysters, indicating a high degree of genetic differentiation between the two groups (average percentage mtDNA nucleotide sequence divergence,=21.8%). The common native haplotype was not present in the selectively inbred sample, and six of the seven haplotypes detected in the inbred oysters were not found in the survey of native oysters. Chi-square tests on haplotype frequencies indicated that the native populations were not significantly different from one another. However, the distribution and relatedness of haplotypes suggest that significant change in the oyster gene pool may have occurred over the past few decades.     

Inter-ocean genetic divergence of istiophorid billfishes

Restriction fragment length polymorphism (RFLP) analysis of mitochondrial DNA (mtDNA) was used to investigate the taxonomic status of the following species-pairs of Atlantic and Indo-Pacific istiophorid billfishes: Atlantic blue marlin Makaira nigricans (Lacépède) and Indo-Pacific blue marlin M. mazara (Jordan and Snyder); Atlantic sailfish Istiophorus albicans (Latreille) and Indo-Pacific sailfish I. platypterus (Shaw and Nodder); and white marlin Tetrapturus albidus Poey and striped marlin T. audax (Phillippi). Tissue samples were collected from 1990 to 1992. Several mtDNA haplotypes were common to Atlantic and Indo-Pacific samples of blue marlin and sailfish, although there were significant differences in the distribution of haplotypes between samples from different oceans. For both blue marlin and sailfish, a single group of closely related mtDNA haplotypes was found among all indo-Pacific and some Atlantic individuals, while the remaining Atlantic specimens exhibited mtDNA haplotypes that differed by several consistent restriction site changes from the common haplotype. No restriction site differences were found to discriminate white marlin from striped marlin, and the mtDNA haplotypes of both species were very similar although significant differences were found in the distribution of haplotypes between the two species. Two of 26 haplotypes were shared between white and striped marlin, and the corrected mean nucleotide sequence divergence between species (0.06%) was not much greater than that observed between geographically distant samples of striped marlin from the Pacific Ocean (mean 0.03%). The presence of identical haplotypes in samples from both oceans for each of the three species-pairs of istiophorid billfishes suggests that specific status may not be warranted for any of the Atlantic and Indo-Pacific populations. The significant difference in the distributions of mtDNA haplotypes between Atlantic and Indo-Pacific populations, which contrasts sharply with the homogeneity reported for several species of tunas, indicates considerable population structuring within the highly vagile billfishes.     

Doubly uniparental transmission of mitochondrial DNA length variants in the mussel <Emphasis Type="Italic">Mytilus</Emphasis><Emphasis Type="Italic">trossulus</Emphasis>

Doubly uniparental inheritance of mitochondrial DNA (mtDNA) is an unusual feature found in marine mussels and a few other bivalve species. In addition to a mitochondrial genome (F) inherited through the female line, heteroplasmic individuals (males) contain a second highly diverged genome (M) that is inherited through the paternal line. The Baltic mussel Mytilus trossulus is characterized by the presence of two phylogenetically close female and male genomes. We examined M. trossulus sampled from the Gulf of Gdansk, southern Baltic. Somatic tissues and gametes were surveyed for the presence of different types of mtDNA. Length variants were identified using PCR (polymerase chain reaction) amplification of the mtDNA fragment containing the major noncoding region. Seventeen length variants present in homo- and heteroplasmic individuals were found in a sample of 343 individuals analyzed. Two length variants of the major noncoding region (PCR product lengths: 1520 and 1370 bp) were the most frequent. Length heteroplasmy was found in 46.8% of males, and was mostly caused by the presence of the short length variant (PCR product length: 1370 bp) of the major noncoding region. MtDNA variants were also detected by restriction analysis of a 1.3 kb segment of coding region (ND2-COIII) amplified by PCR. This study provides evidence that the short length variant is always transmitted to sperm and has taken over the role of the M genome. The remaining length variants were present in both males and females. Longer length variants were transmitted mainly through the female line. A possibility of much higher incidence of genome role reversals in Baltic M. trossulus, in comparison with other populations of mussels is discussed.     

Effects of population-related variation in plant primary and secondary metabolites on aboveground and belowground multitrophic interactions

Insects feeding on aboveground and belowground tissues can influence each other through their shared plant and this is often mediated by changes in plant chemistry. We examined the effects of belowground root fly (Delia radicum) herbivory on the performance of an aboveground herbivore (Plutella xylostella) and its endoparasitoid wasp (Cotesia vestalis). Insects were reared on three populations of wild cabbage (Brassica oleracea) plants, exhibiting qualitative and quantitative differences in root and shoot defense chemistry, that had or had not been exposed to root herbivory. In addition, we measured primary (amino acids and sugars) and secondary [glucosinolate (GS)] chemistry in plants exposed to the various plant population-treatment combinations to determine to what extent plant chemistry could explain variation in insect performance variables using multivariate statistics. In general, insect performance was more strongly affected by plant population than by herbivory in the opposite compartment, suggesting that population-related differences in plant quality are larger than those induced by herbivory. Sugar profiles were similar in the three populations and concentrations only changed in damaged tissues. In addition to population-related differences, amino acid concentrations primarily changed locally in response to herbivory. Whether GS concentrations changed in response to herbivory (indole GS) or whether there were only population-related differences (aliphatic GS) depended on GS class. Poor correlations between performance and chemical attributes made biological interpretation of these results difficult. Moreover, trade-offs between life history traits suggest that factors other than food nutritional quality contribute to the expression of life history traits.     

解淀粉芽孢杆菌DC-4豆豉溶栓酶成熟肽编码序列的克隆及表达

利用PCR方法从解淀粉芽孢杆菌DC 4总DNA中扩增出豆豉溶栓酶 (DFE)成熟肽编码区片段 .测序结果表明 :DFE成熟肽编码区长 82 5bp,编码 2 75个氨基酸残基 ,分子量为 2 7.7× 10 3 ,推导的N’ -端氨基酸序列与豆豉溶栓酶N’ -端氨基酸测序结果完全一致 ,说明克隆到的基因确实是豆豉溶栓酶基因 .同源性分析表明 ,DFE成熟肽编码区的核苷酸和氨基酸序列与日本纳豆激酶的同源性分别为 80 .0 %和 86 .5 % ,这提示豆豉溶栓酶可能是一种新型的溶栓酶 .将表达质粒pET Nde转化E .coliBL2 1(DE3)中 ,IPTG可诱导表达大量的DFE融合蛋白 ,占菌体可溶性蛋白的 4 0 % ,主要以包涵体的形式存在 .图 3表 1参 19     

钼和硼污染对大豆品质的影响

以3个大豆(Glycine max)品种(浙春3号、浙春2号和3811)为材料，设置了4个钼、硼处理(适量钼和硼、高钼、高硼及高钼高硼同施)，研究了钼污染和硼污染对大豆品质的影响．结果表明：高钼或高硼促使大豆种子蛋白质、维生素C、氮、磷、钾的含量显著减少，氨基酸的总量和必需氨基酸的总量大大降低，氨基酸各组分(除脯氨酸外)的含量都明显下降，脂肪的组分也发生变化，对种子中钙镁含量的影响不大，且高硼对大豆品质的毒害作用明显大于高钼．在过量条件下，钼和硼在大豆体内呈现相互抑制作用．3个大豆品种的品质问存在一定的基因型差异，3个品种对高钼和高硼的反应也存在差异．图2表3参18