一个小麦-中间偃麦草异代换系的形态学和细胞学鉴定

中间偃麦草含有丰富的优良基因,在小麦的遗传改良中具有重要利用价值。对从中间偃麦草与小麦品种烟农15杂种后代(BC2F4)中选育的小麦种质系山农0095进行形态学和细胞学鉴定,结果表明：山农0095株高78cm,穗长17．3cm,旗叶长36．3cm,旗叶宽3．03cm,茎杆粗壮,繁茂性好,既长又宽的旗叶、长圆锥型穗是其显著的形态学特征;其根尖细胞染色体数日为2n=42,花粉母细胞减数分裂中期Ⅰ(PMC M Ⅰ)染色体构型为2n=21Ⅱ;它与普通小麦的杂种FⅠPMC M Ⅰ绝大多数细胞出现2个单价体,没有观察到多价体,平均染色体构型为2n=20．08Ⅱ 1．84Ⅰ。以上结果表明,山农0095是一个小麦-中间偃麦草的双体异代换系。     

普通小麦-百萨偃麦草(Thinopyrum bessarabicum)二体异附加系的选育与鉴定

为转移与利用百萨偃麦草耐盐、抗病等优良基因,用普通小麦中国春-百萨偃麦草双倍体与中国春杂交,通过染色体C-分带、分子原位杂交并结合减数分裂中期I的染色体配对分析,从回交后代中选育出一套小麦-百萨偃麦草二体异附加系。对这套异附加系进行的鉴定与分析表明,各附加系除添加了一对百萨偃麦草染色体外,小麦的21对染色体未见明显变化。各附加系所添加的百萨偃麦草染色体在减数分裂中期I配对基本正常,仅有少量单价体,其自交后代中外源染色体亦能正常传递。这说明所培育的这套二体异附加系在细胞学上已相对稳定,暂分别编号为DAJ1、DAJ2、DAJ3、DAJ4、DAJ5、DAJ6和DAJ7。各异附加系中百萨偃麦草染色体在小麦族中的部分同源群归属和百萨偃麦草耐盐抗病基因在染色体上的定位研究正在进行之中。     

小麦-大麦异代换系的创制及鉴定研究

利用"缺体回交法"以小大麦二体异附加系WBA9816作父本与阿勃缺体小麦杂交,创制小大麦异代换系.F1再用该异附加系回交,回交后代通过细胞学鉴定,筛选2n=43的双单体植株套袋自交,从自交后代群体培育出WBS02126;用原位杂交GISH和染色体C-分带技术鉴定表明,WBS02126为2D/2H异代换系;田间试验结果显示,WBS02126生长发育良好,育性基本正常,表现弱春性,叶片宽厚上挺,叶色淡绿,棒状穗,小穗排列紧密,长芒,早熟,综合抗病性好.     

一个多抗小麦—中间偃麦草新种质的选育和细胞分子生物学鉴定

经过多年田间和温室接种抗病性鉴定,从(77-5433×中5)杂交组合花药培养后代中选育出一个兼抗大麦黄矮病、条锈、叶锈和秆锈4种小麦主要病害的新种质遗4212。遗4212的体细胞染色体数为42,在减数分裂中期Ⅰ,在几乎所有的花粉母细胞中都可以观察到21个二价体,这说明遗4212是一个在遗传上业已稳定的整倍体材料。对(遗4212×77-5433)F_1代花粉母细胞的观察表明,遗4212可能是含1对外源中间偃麦草染色体的代换系或具较大中间偃麦草染色体片段的易位系。用基因组原位杂交(genomic in situ hybridization,GISH)对遗4212的有丝分裂中期相、减数分裂后期Ⅰ相和(遗4212×77-5433)F_1代花粉母细胞减数分裂中期Ⅰ、后期Ⅰ进行了检测,确证遗4212含1对外源中间偃麦草染色体。这些结果表明,遗4212是一个小麦一中间偃麦草代换系,其抗病性来自其携带的1对中间偃麦草。     

抗白粉病小麦-中间偃麦草二体异附加系与杀配子材料杂交后代的细胞遗传学研究

利用已选育的抗白粉病烟农15-中间偃麦草二体异附加系与农林26-离果山羊草3C染色体附加系杂交．对其F1、F2、F3的细胞遗传学进行研究．结果表明：F1花粉母细胞减数分裂中期Ⅰ染色体构型紊乱,在39．48％的细胞中发现染色体断片、单价体,后期Ⅰ、后期Ⅱ出现落后染色体、染色体桥．四分体期微核出现频率达48．65％,说明杀配子染色体可有效诱导染色体发生断裂等结构变异;F2代在细胞学方面仍不稳定,表现为染色体数目发生变异,花粉母细胞减数分裂染色体构型紊乱。多价体、落后染色体、染色体桥及微核的普遍出现．说明染色体问可能发生断裂、重接、交换或易位等现象,F2代白粉病抗性也出现分离;F3代虽然染色体数日和白粉病抗性仍在分离,但花粉母细胞减数分裂中期Ⅰ染色体构型较F2稳定,相对紊乱系数下降．从F3代鉴定出了染色体数目为42、构型稳定且对白粉病表现免疫的单株。     

普通小麦—簇毛麦异附加系和异代换系的C—分带鉴定

用改良的C-分带技术鉴定南京农业大学细胞遗传研究室获得的普通小麦的簇毛麦V_2、V_3、V_4、V_6、V_7染色体异附加系和V_2、V_5异代换系,得到与N-分带和染色体配对分析一致的结果,并且由于C-分带可同时鉴别小麦全部21对染色体,鉴定出V_2异代换系中被代换掉的小麦染色体为1A。     

Genomic in situ hybridization (GISH) analyses of Thinopyrum intermedium, its partial amphiploid Zhong 5, and disease-resistant derivatives in wheat

Genomic in situhybridization (GISH) to root-tip cells at mitotic metaphase, using genomic DNA probes from Thinopyrum intermedium and Pseudoroegneria strigosa, was used to examine the genomic constitution of Th. intermedium, the 56-chromosome partial amphiploid to wheat called Zhong 5 and disease-resistant derivatives of Zhong 5, in a wheat background. Evidence from GISH indicated that Th. intermedium contained seven pairs of St, seven J^S and 21 J chromosomes; three pairs of Th. intermedium chromosomes with satellites in their short arms belonging to the St, J, J genomes and homoeologous groups 1, 1, and 5 respectively. GISH results using different materials and different probes showed that seven pairs of added Th. intermedium chromosomes in Zhong 5 included three pairs of St chromosomes, two pairs of J^S chromosomes and two pairs of St-J^S reciprocal tanslocation chromosomes. A pair of chromosomes, which substituted a pair of wheat chromosomes in Yi 4212 and in HG 295 and was added to 21 pairs of wheat chromosomes in the disomic additions Z1, Z2 and Z6, conferred BYDV-resistance and was identical to a pair of St-J^S tanslocation chromosomes (StJ^S) in Zhong 5. The StJ^S chromosome had a special GISH signal pattern and could be easily distinguished from other added chromosomes in Zhong 5; it has not yet been possible to locate the BYDV-resistant gene(s) of this translocated chromosome either in the St chromosome portion belonging to homoeologous group 2 or in the J^S chromosome portion whose homoeologous group relationship is still uncertain. Among 22 chromosome pairs in disomic addition line Z3, the added chromosome pair had satellites and belonged to the St genome and homoeologous group 1. Disomic addition line Z4 carried a pair of added chromosomes which was composed of a group-7 J^S chromosome translocated with a wheat chromosome; this chromosome was different to 7 Ai-1, but was identical to 7 Ai-2. The leaf rust and stem rust resistance genes were located in the distal region of the long arm, whereas the stripe rust resistance gene(s) was located in the short arm or in the proximal region of the long arm of 7 Ai-2. A pair of J^S-wheat translocation chromosomes, which originated from the WJ^S chromosomes in Z4, was added to the disomic addition line Z5; the added chromosomes of Z5 carried leaf and stem rust resistance but not stripe rust resistance; Z5 is a potentially useful source for rust resistance genes in wheat breeding and for cloning these novel rust-resistant genes. GISH analysis using the St genome as a probe has proved advantageous in identifying alien Th. intermedium in wheat. Received: 17 May 1999 / Accepted: 22 June 1999     

OsBTF3过表达转基因水稻株系的创制及其分子鉴定

本研究旨在创制OsBTF3过表达转基因水稻株系,为验证OsBTF3基因在水稻抗性和生长发育中的功能、评价其在水稻农艺性状遗传改良中的应用价值提供试验材料。通过基因过表达载体构建、水稻愈伤组织诱导、农杆菌介导愈伤组织转化、植株再生、潮霉素抗性(Hyg^R)筛选及PCR验证、基因过表达RT-Q-PCR检测等方法,成功地获得了97个T₀代和20个T₁代过表达转基因株系,并分别得到分子验证。与野生型对照株相比,5个T₁代过表达株系中的OsBTF3基因表达水平显著提高,平均高达3.58倍。因此,由组成型表达的35S启动子驱动的OsBTF3基因在转基因水稻株系中成功地得到了增量表达,并对水稻生长发育、抗病性和抗逆性具有调控作用。     

小粒野生稻导入系对稻飞虱的抗性鉴定与分析

稻飞虱是水稻生产最严重的害虫之一。野生稻拥有丰富的抗虫基因资源,导入系是鉴定和利用野生稻有利基因的有效途径。本研究通过对371份小粒野生稻导入系进行抗褐飞虱和白背飞虱接虫鉴定,分别筛选出了11份抗、72份中抗褐飞虱的材料和7份抗、45份中抗白背飞虱的材料,其中有5份材料兼抗褐飞虱和白背飞虱,这是从小粒野生稻中鉴定出抗白背飞虱材料的首次报道。通过对2份抗性导入系材料与感虫亲本杂交构建的F1和F2群体的抗虫鉴定和分析表明:K41对褐飞虱和白背飞虱的抗性受2对显性抗虫基因通过互补作用所控制;P114对褐飞虱和白背飞虱的抗性都是由1对主效的隐性基因控制。这些结果必将有利于小粒野生稻抗稻飞虱的基因定位和育种利用。     

基于单片段代换系的水稻穗长QTL加性及其上位性效应

穗长是影响水稻(Oryza sativa)产量的重要因子之一, 研究水稻穗长QTL间的上位性效应对于发掘水稻产量潜力具有重要意义。该研究以16个单片段代换系(single segment substitution lines, SSSLs)和15个双片段聚合系(double segment pyramiding lines, DSPLs)为材料研究了水稻穗长QTL的加性及上位性效应。以P<0.01为阈值, 共检测到6个穗长QTL和9对基因互作座位。其中2个(Pl-2和Pl-10)是尚未报道的穗长QTL。穗长QTL互作后, 一些互作对的上位性效应与单个QTL的作用方式及效应大小各不相同, 预示着基因聚合后会产生不同的互作效应。该研究结果对于通过分子聚合育种手段改良穗长具有重要意义。     

白木香内生真菌的分离及分子鉴定

对采自广东省信宜市的白木香进行了内生真菌分离培养,共获得50个菌株,通过rDNA中内转录间隔区(ITS)序列鉴定其为20个种,其中确定至种名的有7属8种,仅确定至属名的有8种,不能确定种属的有4种,其中A20菌株(Fimetariella rabenhorstii)为国内首次报道。刺盘孢菌(Colletotrichum)共分离到17株,占分离总数的34%,为白木香优势种群;镰刀菌(Fusarium)共分离到11株,是结香部位的优势种群。白木香内生真菌分布部位的专一性不明显,茎中分离的数量及种类最多,叶中最少;5年龄内生真菌的多样性较好,10月龄的多样性较低。     

分子生物学技术在植物内生菌分类鉴定中的应用

植物内生菌作为微生物中的一个研究领域,近年来一直备受关注。不仅是因为内生菌在农牧业生产和环境净化方面应用的良好表现,而且还为新药研制提供了新的途径。与此同时,植物内生菌分类鉴定的研究也有一定的进展,特别是在分子生物学技术应用到其中后。基于这点,就最近10年来有关植物内生菌分类鉴定的文献所使用的技术进行了分析和讨论。     

K型小麦细胞质雄性不育保持系T911289中外源遗传物质的初步鉴定

利用荧光基因组原位杂交(GISH)、生化标记和DNA分子标记技术对普通小麦(triticum aestivum L.)K型细胞质雄性不育保持系T911289的染色体组成进行了鉴定与分析。GISH鉴定和黑麦特异散布重复序列的检测结果表明,T911289的外源遗传物质来源于黑麦,黑麦1RS上的微卫星引物SCM9扩增结果和醇溶蛋白酸性聚丙烯酰胺凝胶电泳(A—PAGE)分析、低分子量谷蛋白的sDS_PAGE分析均表明,T911289所含的黑麦遗传物质来源于1RS;A-PAGE和SDS-PAGE分析及小麦1BS上的微卫星引物的扩增结果则表明,‘1911289缺少1BS染色体臂或1BS末端片段。GISH鉴定结果还表明,‘1911289中有罗泊逊易位和小片段易位两种类型的杂交信号,说明T911289是一个异质群体,但其罗泊逊易位又不同于生产上大面积应用的1BL／1RS易位,它可能是一种新的复杂易位形式。虽然T911289的小片段易位未能打破优异农艺性状与劣质蛋白基因的连锁,但这种小片段易位的获得将有利于小麦和黑麦的遗传研究,这种种质材料在育种上的应用价值也应优于罗泊逊易位。     

两株滇产广义美味牛肝菌的分离培养及其分子鉴定

采用组织分离法从广义美味牛肝菌子实体分离获得2株稳定的菌株,初步研究了两菌株的分离和培养条件。用ITS序列分析,对分离菌株进行了分子鉴定。基于ITS序列构建的系统树表明两株菌属于美味牛肝菌复合群,并与夏生牛肝菌Boletus aestivalis（Paul.）Fr.有较近的亲缘关系。     

Development of Chromosome Segment Substitution Lines Derived from Backcross between Two Sequenced Rice Cultivars, <Emphasis Type="Italic">Indica</Emphasis> Recipient 93-11 and <Emphasis Type="Italic">Japonica</Emphasis> Donor Nipponbare

Chromosome segment substitution lines (CSSLs) are powerful tools for detecting and precisely mapping quantitative trait loci (QTLs) and evaluating gene action as a single factor. In this study, 103 CSSLs were produced using two sequenced rice cultivars: 93-11, an elite restorer indica cultivar as recipient, and Nipponbare, a japonica cultivar, as donor. Each CSSL carried a single chromosome substituted segment. The total length of the substituted segments in the CSSLs was 2,590.6 cM, which was 1.7 times of the rice genome. To evaluate the potential application of these CSSLs for QTL detection, phenotypic variations of seed shattering, grain length and grain width in 10 CSSLs were observed. Two QTLs for seed shattering and three for grain length and grain width were identified and mapped on rice chromosomes. The results demonstrate that CSSLs are excellent genetic materials for dissecting complex traits into a set of monogenic loci. These CSSLs are of great potential value for QTL mapping and plant marker-assisted breeding (MAB).     

Occurrence and Molecular Identification of Anisakis Dujardin, 1845 from Marine Fish in Southern Makassar Strait,Indonesia

Anisakis spp. (Nematoda: Anisakidae) parasitize a wide range of marine animals, mammals serving as the definitive host and different fish species as intermediate or paratenic hosts. In this study, 18 fish species were investigated for Anisakis infection. Katsuwonus pelamis, Euthynnus affinis, Caranx sp., and Auxis thazard were infected with high prevalence of Anisakis type I, while Cephalopholis cyanostigma and Rastrelliger kanagurta revealed low prevalence. The mean intensity of Anisakis larvae in K. pelamis and A. thazard was 49.7 and 5.6, respectively. A total of 73 Anisakis type I larvae collected from K. pelamis and A. thazard were all identified as Anisakis typica by PCR-RFLP analysis. Five specimens of Anisakis from K. pelamis and 15 specimens from A. thazard were sequenced using ITS1-5.8S-ITS2 region and 6 specimens from A. thazard and 4 specimens from K. pelamis were sequenced in mtDNA cox2 region. Alignments of the samples in the ITS region showed 2 patterns of nucleotides. The first pattern (genotype) of Anisakis from A. thazard had 100% similarity with adult A. typica from dolphins from USA, whereas the second genotype from A. thazard and K. pelamis had 4 base pairs different in ITS1 region with adult A. typica from USA. In the mtDNA cox2 regions, Anisakis type I specimens from A. thazard and K. pelamis showed similarity range from 94% to 99% with A. typica {"type":"entrez-nucleotide","attrs":{"text":"AB517571","term_id":"283379497","term_text":"AB517571"}}AB517571/{"type":"entrez-nucleotide","attrs":{"text":"DQ116427","term_id":"74229655","term_text":"DQ116427"}}DQ116427. The difference of 4 bp nucleotides in ITS1 regions and divergence into 2 subgroups in mtDNA cox2 indicating the existence of A. typica sibling species in the Makassar Strait.     

Molecular Identification of Diphyllobothrium nihonkaiense from 3 Human Cases in Heilongjiang Province with a Brief Literature Review in China

Human diphyllobothriasis is a widespread fish-borne zoonosis caused by the infection with broad tapeworms belonging to the genus Diphyllobothrium. In mainland China, so far 20 human cases of Diphyllobothrium infections have been reported, and the etiologic species were identified as D. latum and D. nihonkaiense based on morphological characteristics or molecular analysis. In the present study, proglottids of diphyllobothriid tapeworms from 3 human cases that occurred in Heilongjiang Province, China were identified as D. nihonkaiense by sequencing mitochondrial cytochrome c oxidase subunit I (cox1) and NADH dehydrogenase subunit 5 (nad5) genes. Two different cox1 gene sequences were obtained. One sequence showed 100% homology with those from humans in Japan. The remaining cox1 gene sequence and 2 different nad5 gene sequences obtained were not described previously, and might reflect endemic genetic characterizations. D. nihonkaiense might also be a major causative species of human diphyllobothriasis in China. Meanwhile, the finding of the first pediatric case of D. nihonkaiense infection in China suggests that infants infected with D. nihonkaiense should not be ignored.     

内蒙古地区白桦外生菌根形态类型及分子鉴定

以内蒙古不同地区白桦外生菌根为材料,采用形态解剖学方法和分子生物学手段对与白桦共生的外生菌根真菌多样性进行全面的调查,并经对所测序列与GenBank和Database比对。结果显示:在内蒙古地区与白桦共生的外生菌根真菌共13种,其中担子菌7种,子囊菌4种,分别来自于丝膜菌属、丝盖伞属、蜡壳耳属、毛革菌属、滑菇属和空团菌属、块菌属、地怀菌属。其中,菌根类型T₈和T₁₁未能提出其总DNA,根据其外生菌根形态类型并参照Agerer体视显微镜菌根图谱和Haug菌根图谱进行比较,分类鉴定为荷顿氏疣柄牛肝菌和白桦外生菌根真菌一种。结果表明,内蒙古地区白桦外生菌根真菌多样性相对较高,且与利用地上子实体鉴定的外生菌根真菌种类有一定的区别。