用同工酶和RAPD技术研究棉花三元杂种石远321新品种的遗传特性

石远321是从海岛棉×野生色伯氏棉×陆地棉三元杂种后代中选育而成的种间杂交新品种。其细胞学鉴定结果、形态学基本特征以及栽培特性均为陆地棉型。但在丰产性、棉纤维品 质及抗逆性等许多农艺性状和经济性状方面,该品种明显综合了多棉种多亲本的遗传特性。为了从分子水平上认识石远321的三元杂种性质及其变异水平,用等电聚焦电泳和RAPD技术,对石远321及其不同棉种亲本进行了遗传分析。主要结果如下（1）酯酶同工酶图谱分析表明,石远321与其野生棉亲本和海岛棉亲本同具有一条陆地棉亲本所没有的特异带;（2）用4个引物对石远321扩增共发现6个野生棉亲本和海岛棉亲本的特征片段;（3）通过比较石远321与其3个不同棉种亲本之间的RAPD差异,说明石远321在基因组水平上具有高度的遗传异质性。初步从分子水平上证实了石远321为上述3个棉种的种间杂交后代。     

陆地棉-野生比克氏棉杂种红花种质系的育成

采用种间杂交新技术,得到了陆地棉×野生比克氏棉的F_1不育株,在嫁接教养及短日凉爽条件下培育,雌雄恢复了低育性,通过3次回交及对早期剧烈分离世代的选择之后,首次育成了兼具双亲特征,即既有比克氏棉的红花紫花斑及细强纤维,又有陆地棉的株型、熟期、白绒、高衣分和结铃性的稳定的杂种选系,命名为“HB红花系”,这是一种前所未有的新型种质资源,具有新品种选育及遗传研究的重要价值.     

用RAPD标记评估我国棉花品种遗传多样性

利用ＲＡＰＤ标记对１７个不同来源的我国棉花品种（系）进行遗传多样性评估。２００个随机引物扩增出１１３条多态性片段。１７个品种（系）依据染色体组的不同可以分为３大类。它们之间遗传相似性系数从０．２０００到０．８４５１。３个大面积推广品种鄂荆１号,中棉所１２号和苏棉３号遗传基础十分相近。陆地棉与阔叶棉杂交产生的野生种质系７５８１,与其他陆地棉品种（系）遗传差异显著。不同育种单位在育种过程中种质使用具有     

栽培灯盏花DNA指纹图谱研究及遗传多样性分析

目的:采用ISSR标记构建云南省栽培灯盏花DNA指纹图谱并进行遗传多样性分析.方法:从20对候选引物中筛选出7对引物对云南10个地方栽培灯盏花进行扩增,通过扩增带型的差异构建其DNA指纹图谱,利用Popgen32、NTSYS2软件进行遗传一致性系数和遗传多样分析.结果:7对引物在10份共扩增出48条谱带,其中33条具有多态性,占68.8％,表明各居群的遗传差异较大.10个地方种质资源被聚为2个大类,野生居群被聚为一类,栽培灯盏花聚为一类,其中栽培灯盏花有两大亚类.结论:通过构建DNA指纹图谱容易地把灯盏花种植资源相互区分鉴别出来,10个地方灯盏花种质资源遗传多样性丰富,栽培灯盏花与野生具有差异,栽培种质材料之间也有差异.     

向陆地棉渐渗野生比克氏棉腺体延缓形成基因的方法研究

以隐性无腺体陆地棉作母本,比克氏棉作父本杂交,获得了三倍体杂种［（ＡＤ）１Ｇ１］,该杂种完全表达了比克氏棉的腺体延缓形成基因（Ｇｌ＾ｂｉｃ）的特征,Ｆ９胚无腺体,Ｆ１植株有腺体。位于Ｇ１染色体组上的腺体延缓形成基因Ｇｌ＾ｂｉｃ对陆地棉（ＡＤ）１上的无腺体基因（ｇｌ２ｇｌ３）表现为显性,对正常腺体基因Ｇｌ２Ｇｌ３则表现为隐性。探讨了向陆地棉渐渗野生棉腺体延缓形成基因的方法。     

5个澳洲野生棉种植株形态性状与叶片过氧化物同工酶谱研究

研究了斯特提棉、南岱华棉、澳洲棉、纳尔逊氏棉和比克氏棉５个澳洲野生棉各的种卫和植株的形态性状、色素腺体性状、棉酚含量和叶片过氧化物同工酶等主要主要性状。结果表明,斯特提棉和南岱华棉、澳洲棉和纳尔逊氏的种子、植株和花器形态性状,以及叶片过氧化物同工酶谱等性状分别比较相似,经克氏棉的形态特征介上述两类棉种之间,略近似于澳洲棉和纳尔逊氏棉,５个棉种均具有种子地色素腺体植株有色素腺体的特性,其体眠种子的棉     

棉属种间高代系的过氧化物酶同工酶研究

利用薄层等电聚焦方法,对已稳定遗传２０代的棉属陆地棉（Ｇｏｓｓｙｐｉｕｍ ｈｉｒｓｕｔｕｍ ）×中棉（Ｇ．ａｒｂｏｒｅｕｍ ）种间高代系蕾期真叶及花药中的过氧化物酶同工酶进行研究,结果表明：１．同一棉种的不同品种,其过氧化物酶同工酶基本相同,同一染色体组的不同棉种,其过氧化物酶同工酶存在一定的差异,不同染色体组的棉种,其过氧化物酶同工酶存在显著的差异;２．高代系的同工酶谱与母本“科遗２号”相似,而与父本中棉“完紫”具有较大的差异     

四倍体大燕麦×六倍体裸燕麦的杂种F1的产生及鉴定

本研究以四倍体大燕麦（Avena magna L.）做母本,六倍体裸燕麦（Avena nuda L.）做父本进行杂交,利用幼胚拯救技术获得了杂种F1,并对其后代形态特征进行了观察;对杂种F1同工酶图谱和DNA指纹图谱进行了分析。杂种F1形态特征偏亲本或介于双亲之间;同工酶研究表明多数F1具有双亲互补酶带;RAPD分析不同引物扩增产物F1呈共显性或偏父、偏母。这些结果表明F1为真杂种。     

水生实验动物剑尾鱼的DNA指纹图谱构建

目的剑尾鱼是由原良种委员会审定并由农业部公布的水生实验动物,在遗传学研究、水环境污染监测、细菌性疾病研究方面显示出较好的应用前景。为了对剑尾鱼选育系进行种质资源监测、区分选育系与非选育以及鉴定近交系纯度,本研究采用微卫星DNA进行水生实验动物剑尾鱼的指纹图谱构建。方法根据相关报道设计合成了50对微卫星引物,对几个剑尾鱼品系的种质资源进行检测,筛选品系间差异性引物;确立剑尾鱼核心引物,用EXCEL散点图绘制DNA指纹图谱模式图;并将数字化指纹数据输入珠江水产研究所鱼类种质鉴定软件V1.0,形成剑尾鱼标准化指纹图谱鉴定数据库。结果共获得剑尾鱼品系间特异标记5个可用于剑尾鱼近交系鉴定,确立46个微卫星标记为核心引物,构建剑尾鱼选育系RR-B系、RW-H系和非选育的野生品种的DNA指纹图谱。结论本研究筛选出的微卫星标记与构建的指纹图谱,可用于剑尾鱼3个品种间的品种鉴定、纯度检测及遗传监测。     

双孢蘑菇杂交菌株As2796家系的分子遗传研究

应用PCR和凝胶电泳等技术,对双孢蘑菇杂交菌株As2796及其亲本和子代作分子遗传标记跟踪分析,结果如下：1）总DNA的RAPD分析表明,随着遗传代数的增加,杂种子代和出发异核体亲本间的遗传差异逐渐增大;2）mtDNA的酶切图谱表明,亲本8213及其杂交子代具有相同的基因型,表明双孢蘑菇mtDNA呈单亲遗传;3）Est同工酶的PAGE图谱表明,结合了亲本02高产特征和8213优质特征的杂交子代具有两个亲本的标记带型,证明Est同工酶标记是双孢蘑菇新菌株特性预测或鉴定的有效指标。     

RAPD标记在紫菜遗传多样性检测和种质鉴定中的应用

用ＲＡＰＤ技术对４类紫菜（Ｐｏｒｐｈｙｒａ ｙｅｚｏｅｎｓｉｓ,Ｐ．ｈａｉｔａｎｅｎｓｉｓ,Ｐ．ｋａｔａｄｎｉ ｖａｒ．ｈｅｍｉｐｈｙｌｌａ和Ｐ．ｏｌｉｇｏｓｐｅｒｍａｔａｎｇｉａ）的１５个无性系丝状体进行了遗传多样履分析,从５０个ＯＰＥＲＯＮ引物中经过初筛,其中６个引物可以扩增出稳定的可重复的图谱。这６个引物共扩增出了６０条带,多态性比例达９７．１％。根据ＲＡＰＤ结果将这１５个无性了紫菜的ＤＮＡ     

New red flower germplasm lines of cotton selected from hybrid of Gossypium hirsutum × G. bickii

By means of dropping GA3 (50 ppm) and NAA (40 ppm) on the hybrid boll-embryo culture in vitro, one F1 plant of G. hirsutum G. bickii was obtained; when F1 branches were grafted on upland cotton and then back-crossed with upland cotton under short-day and cooler-night condition, some BC1 seeds could be harvested. The characteristic segregation was very violent in early generation. Through 3 times of back-crossing and selecting, ten stable hybrid lines with the character of both male parent (viz. red petal-purple spot and strong fibre) and female par-ent (plant type, earliness, white fibre, lint length, etc. ) were established. These lines were assigned as HB red flow-er lines (HBRL). Transference of character of G. bickii to upland cotton was proved to be successful for the first time. These new germplasms may play an important role in both the genetic research and new cotton variety breeding.     

Molecular characterization of a new type of cytoplasmic male sterile sugar beet

A line (named Cl) of cytoplasmic sterility of sugar beet whose cytoplasm derived from Betacicla Turkey was obtained by interspecific hybrid. Its cytoplasm and a spontaneous male sterile cytoplasm from wild beet Beta maritima (named M) were compared with that of Owen's sterile line (S-cms) and a common maintainer of them named N was used as control. RFLP and RAPD methods were mainly used in our experiments. The restriction fragment patterns of mtDNAs were found to be likely but for a few of specific low-lighted electrophoresis bands in Cl. The results of Southern hybridization of six heterogeneous mitochondrial genes as probes to digests of mtDNAs by six restriction enzymes showed to be analogous between S and M lines. But the Cl mtDNA was sorted out by hybridization of atpA probe. Difference of low-molecular-weight mitochondrial DNAs was found among the three sterile lines. Three RNA molecules weighing about 4.2kb stably existed in Cl mitochondria. Our results of RAPD also supported that the Cl cytoplas     

Production of somatic hybrids between Diospyros glandulosa and D. kaki by protoplast fusion

Interspecific somatic hybrids between Diospyros glandulosa (2n=2x=30) and D. kaki cv. Jiro (2n=6x=90) were produced by electrofusion of protoplasts. Protoplasts were isolated from calli derived from leaf primordia, fused electrically, and cultured by agarose-bead culture using a modified KM8p medium. Flow cytometry revealed that the nuclear DNA content was the sum of those of D. glandulosa and D. kaki cv. Jiro in 149 of the 166 calli obtained. RAPD analysis showed that the 149 callus lines yielded specific bands for both D. glandulosa and D. kaki cv. Jiro and further confirmed that they were interspecific somatic hybrid calluses. Shoots were regenerated from 63 of the 149 interspecific hybrid calluses. Chloroplast DNA analysis by PCR-RFLP, flow cytometric determination of nuclear DNA content, and RAPD analysis revealed that the 63 interspecific hybrid shoot lines contained the nuclear genomes from both parents but only the chloroplast genome from D. glandulosa. Microscopic observation of root tip cells demonstrated that somatic chromosome number of the interspecific hybrids was 2n=8x=120. This revised version was published online in June 2006 with corrections to the Cover Date.     

Comparison of RFLP and RAPD markers to estimating genetic relationships within and among cruciferous species

Restriction fragment length polymorphism (RFLP) and random amplified polymorphic DNA (RAPD) markers are being used widely for evaluating genetic relationships of crop germplasm. Differences in the properties of these two markers could result in different estimates of genetic relationships among some accessions. Nuclear RFLP markers detected by genomic DNA and cDNA clones and RAPD markers were compared for evaluating genetic relationships among 18 accessions from six cultivated Brassica species and one accession from Raphanus sativus. Based on comparisons of genetic-similarity matrices and cophenetic values, RAPD markers were very similar to RFLP markers for estimating intraspecific genetic relationships; however, the two marker types gave different results for interspecific genetic relationships. The presence of amplified mitochondrial and chloroplast DNA fragments in the RAPD data set did not appear to account for differences in RAPD- and RFLP-based dendrograms. However, hybridization tests of RAPD fragments with similar molecular weights demonstrated that some fragments, scored as identical, were not homologous. In all these cases, the differences occurred at the interspecific level. Our results suggest that RAPD data may be less reliable than RFLP data when estimating genetic relationships of accessions from more than one species.     

粘类小麦细胞质雄性不育系的RAPD分析

利用250条10-聚寡核苷酸随机引物对具粘果山羊草(Aegilops kotschyi)、易变山羊草(Ae.variabilis)、偏凸山羊草(Ae.ventricosa)和二角山羊草(Ae．bicornis)细胞质不育系及其保持系5-1的总DNA进行了RAPD多态性分析,其中31条引物对4种不育系及其保持系总DNA均无扩增,217条引物扩增条带完全相同。有2条随机引物在2种不育系之间有特异的扩增片段,其中引物S22在偏凸山羊草细胞质雄性不育系基因组DNA中扩增出分子量约为1600bp的特异带,引物S202在粘果山羊草细胞质雄性不育系基因组DNA中扩增出约1300bp特异带。线粒体基因组DNA的RAPD分析表明,4种不育系及其保持系mtDNA存在明显的差异。证明了S22—1600为偏凸山羊草细胞质不育系及其mtDNA基因组DNA的RAPD特异片段．S202—1300可能为粘果山羊草细胞质不育系及其ctDNA基因组DNA的RAPD特异片段。     

四个栽培棉种间的杂种F1细胞遗传学与亲缘关系研究

以棉属四个栽培棉种进行种间杂交,产生(亚洲棉×草棉)和(陆地棉×海岛棉)2个二元杂种F1及其[(亚洲棉×草棉)×(陆地棉×海岛棉)]四元杂种F1,观察和测定4个栽培棉种及其2个二元杂种F1和四元杂种F1的花粉母细胞(PMC)减数分裂的染色体行为及其花粉生活力,以研究4个栽培棉种间的亲缘关系和进化关系。结果表明,二元杂种(亚洲棉×草棉)F1的PMC减数分裂中期Ⅰ出现一个四体环,其余为二价体,染色体构型为2n=26=11Ⅱ 1Ⅳ;花粉生活力的测定表明,(亚洲棉×草棉)F1可育型花粉为50.71%,表现为典型的配子半不育特性,说明两个二倍体棉种间发生一次染色体易位。(陆地棉×海岛棉)F1以26个二价体细胞为主,但有少量的单价体、三价体以及四价体,染色体构型为2n=52=0.78Ⅰ 22.24Ⅱ 0.94Ⅲ 0.98Ⅳ。花粉生活力的测定表明,(陆地棉×海岛棉)F1可育型花粉为54.84%,可见2个四倍体棉种间亲缘关系较近,二者之间仅发生了染色体的易位或倒位。而由4个栽培种合成的四元杂种F1,其减数分裂异常,染色体丢失现象普遍,部分染色体不能联会配对,以单价体的形式存在,并出现三价体、四价体、五价体等多价体,染色体构型为2n=52=5.45Ⅰ 14.41Ⅱ 2.44Ⅲ 1.59Ⅳ 0.63Ⅴ 0.15Ⅵ,其可育花粉为6.87%。研究结果表明了4种栽培棉种之间的亲缘关系相对较近,可以通过遗传重组产生综合有4个栽培棉种性状的新种质。     

拟南芥抗盐突变体的RAPD分析

以筛选得到可以稳定遗传的抗盐单基因突变体2^#和15^#以及野生型拟南芥为材料进行RAPD分析,150条引物中有3条引物在突变体之间扩增出多态性,不仅证明了DNA水平突变的发生,而且表明它们之间遗传背景相似,是一系列抗盐性不同的近似等位基因系。1条引物在突变体的扩增产物比在野生型的扩增产物多出一个大小约为1200bp的片段,初步认为该片段与抗盐的主效基因有关。     

川西北荞麦种间亲缘关系初步研究

采用RAPD和同工酶技术对川西北具有代表性的10份荞麦材料进行分析.结果表明,筛选出的15个RAPD引物扩增出388条带.其中352条具有多态性,多态性比率为90.72%;过氧化物同工酶分析获得15条酶带,酯酶同工酶获得10条酶带.3种方法聚类结果基本一致,10份材料可初步聚为4个类群.其中,与金荞亲缘关系相比,甜荞较近而苦荞较远;普格县野荞和齿翅野荞、细柄野荞亲缘关系较近,划为同类;阿坝野荞单独划为一类,对其在分类中的地位有待进一步研究.