花生的荧光显带和rDNA荧光原位杂交核型分析

建立花生准确而详细的核型对于阐明其起源和开展其基因组研究十分重要。本研究采用DAPI显带和5S、45S rDNA探针双色荧光原位杂交对花生有丝分裂中期染色体进行了分析。结果表明,花生的单倍基因组总长度为(81.06±3.74) μm,最长染色体为(4.72±0.15) μm,最短染色体为(2.62±0.14)μm;有15对染色体显示了着丝粒区DAPI⁺带,其中10对为强带,5对为弱带;有2对5S rDNA位点和5对45S rDNA位点,其中1对5S与1对45S位点同线。综合染色体测量数据、DAPI⁺带和rDNA杂交信号,对花生染色体进行了准确配对和排列,建立了详细的分子细胞遗传学核型。花生的核型公式为2n=4x=40=38m+2sm(SAT),核型不对称类型属于2A型。     

莪术CPD染色和45S rDNA荧光原位杂交核型分析

为了对莪术[Curcuma zedoaria (Christm.) Roscoe]的染色体进行识别并对该物种基因组的结构及进化进行初步研究,利用改进的火焰干燥法及荧光原位杂交技术,对莪术中期染色体的长度,着丝粒的位置及随体的数目进行分析。PI和DAPI组合（CPD）染色后和相继的45S rDNA探针荧光原位杂交结果显示,莪术具有五对45S rDNA位点,三对位于8,22,31号染色体末端的CPD带区,二对位于4,30号染色体的短臂上。第五号短臂为富含GC对的非45S rDNA位点。该实验建立了莪术的经典核型,为非整倍体,核型公式为2n＝62+1=40m+12sm+1m,其核型不对称性为2A型。     

葱的CPD染色和45S rDNA FISH核型分析

为给葱的染色体的识别提供新标记,建立葱的分子细胞遗传学核型,本研究采用去壁火焰干燥法制备了分散且形态良好的葱中期染色体,并进行了CPD（PI和DAPI组合）染色和45S rDNA荧光原位杂交（FISH）,根据葱染色体的形态特征,结合CPD染色和FISH结果,对葱进行了核型分析。CPD染色结果:葱所有染色体臂末端都显示CPD带。FISH结果：有一对45S rDNA位点（在第5对染色体上）。葱的核型公式：2n=2x=16=2sm+12m+2st(SAT)。研究表明：利用CPD染色和45S rDNA FISH,不仅能为染色体识别提供新标记,还能了解染色体GC丰富区的分布,为葱属植物的物种鉴定、系统分类与进化等研究提供DNA分子方面的证据。     

叶绿体5S rDNA ITS和matK基因序列应用于刚竹属植物系统分类的可行性分析

本文应用CTAB法从37种竹类植物中提取基因组DNA,根据在GenBank中发表的5S rDNAITS和matK基因设计引物,通过PCR进行扩增.结果表明:37种竹子5S rDNA ITS序列PCR产物大小约为450 bp,在刚竹属内部无长度上的差异,但是舒竹(Phyllostachys shuchengensis)与其他刚竹属植物相比,有一个位点的差异(由A变为C).因此,5S rDNA ITS在属下水平上无法提供较大的信息量,变异性较低,不适于刚竹属属下水平的系统分类研究.同样,选取37种竹子中3个竹种的,matK基因的PCR扩增产物直接进行测序,结果显示marK基因在竹亚科的属间长度上无差异,产物的长度约为1 500 bp,将测序结果进行同源性比对,在变异位点附近寻找多态性酶切位点,碱基序列上有一个位点的差异(BstN Ⅰ).PCR-RFLP结果显示,共有3种竹子在此位点发生变异分别为:浙江淡竹(Phyllostachys meyeri)、安吉金竹(Phyllostachysparvifolia)和黄古竹(Phyllostachys angusta).刚竹属植物的mark基因序列相当保守,片段中刚竹属间的绝对核苷酸差异不到1个,所提供的信息量不够充分.因此,叶绿体5S rDNA ITS和marK基因序列不适用于刚竹属植物系统分类研究,但其可能适合在属或属以上分类等级竹类植物的系统分类中应用.     

从核糖体DNA ITS区序列研究甘蔗属及其近缘属种的系统发育关系

测定了甘蔗属及其近缘属的13个种共43个个体和1个橡草(Pennisetum schumach)外群的核糖体DNA中的内转录间隔区(ITS)及5.8S rDNA 基因的序列.结果表明:甘蔗属及其近缘属种的ITS区(含ITS1,5.8S rDNA和ITS2)序列的长度范围为589～591bp,变异位点为140个,信息位点为60个;其中,ITS1和ITS2的长度范围分别为205～208bp和216～220bp,     

植物孢子体型自交不亲和的分子研究进展

为避免自生花粉的受精导致物种退化,开花植物进化出了许多的机制,自交不亲和系统是其中的一种,可增强植物的多样性以及对自然环境的适应能力,有效促进杂交。根据不同的遗传方式可分为不同类型。本综述以近年来孢子体自交不亲和的研究结果为基础,综述了孢子体型自交不亲和的分子机制,芸薹属及芸薹属以外的植物中S位点与S位点相关基因的功能。除了芸薹属植物以外的其他植物中对孢子体型自交不亲和的研究甚是少见,缺少全面的认识和研究,且信号传递路径还不是完全清楚,还需要进一步的研究。     

45S rDNA基因在新麦草染色体上的分布

分析rDNA基因位点在染色体上的分布可以对新麦草染色体进行识别和分析其基因组特征。利用FISH和顺序C-分带-FISH技术将45S rDNA定位于新麦草细胞分裂中期染色体上,结果表明,45S rDNA在二倍体新麦草染色体上有6个主要分布位点,另外几条染色体在两臂中部或长臂末端还显示出较弱的杂交信号,信号强度显示蒙农4号新麦草基因组具有一定杂合性。分析确定新麦草的45S rDNA基因主位点分别位于N1染色体短臂末端、N3染色体短臂末端以及N5染色体短臂末端,推测这3对染色体是NOR染色体。     

棉花基因组重复序列研究进展

本文对棉族基因组中DNA重复序列的划分、特点进行了概括，介绍了重复序列在基因组的作用和棉花重复序列18S—26S rDNA、5SrDNA的同步进化，并对重复序列在起源和进化、分子标记辅助选择及染色体操作中的应用进行了展望。     

乳酸粪肠球菌的分离鉴定与系统进化分析

目的 分离鉴定一株疑似乳酸粪肠球菌;方法 通过对表型特性包括培养特性、形态学观察、生化试验等鉴定,同时结合16S rDNA系统进化分析;结果：该株革兰氏阳性菌能在10℃和45℃生长,接触酶阴性,发酵葡萄糖产酸不产气,可生长于6.5%NaCl和PH9.6的环境等,16S rDNA系统进化分析与粪肠球菌有最高同源性;结论 分离株为乳酸粪肠球菌。     

小麦的起源、进化与中国小麦遗传资源

本文从小麦属的分类、小麦的近缘植物、小麦的地理起源、小麦栽培起源与传播、小麦的进化、中国小麦遗传资源等方面进行了阐述,全面分析了小麦的起源、进化及可利用资源,为小麦种质创新和培育新品种提供了可靠资料。     

河南麦田野燕麦病原真菌资源调查及致病性测定

为了解中国河南省小麦田常见杂草野燕麦上的病原真菌种类,以期为更好的利用植物病原真菌资源和开发生物除草剂奠定基础。采用组织分离法对野燕麦病害样品进行分离,结合形态观察和rDNA ITS序列分析对菌株进行鉴定,最后用人工接种法对部分菌株进行致病性测定。从81份自然发病样品中分离到157个菌株,共鉴定了9个属的17种真菌。对于做致病性测定的28个菌株,其中新月弯孢菌株YM1362对供试杂草表现出很强致病性,而对棉花、大豆、花生等植物没有致病性。结果表明野燕麦上的病原真菌主要为链格孢属、弯孢属、平脐蠕孢属和凸脐蠕孢属;其中菌株YM1362具有开发为防治双子叶作物田中禾本科杂草的生物除草剂的潜力。     

二倍体野生棉与四倍体栽培棉基于GISH的遗传关系分析

[目的]研究二倍体野生棉与四倍体栽培棉间的遗传亲缘关系,进一步探索各棉种间的起源与进化.[方法]以5个二倍体基因组的代表种B1(异常棉)、C1(斯特提棉)、E2(索马里棉)、F1(长萼棉)以及G1(比克氏棉)的基因组DNA(gDNA)为探针,以2个四倍体栽培种(陆地棉中棉所16、海岛棉新海7号)有丝分裂中期染色体为靶DNA,进行了基因组原位杂交(Genomic in situ hybridization,GISH)分析.[结果]以B1、E2和F1gDNA为探针时,杂交信号主要分布在2个栽培种较长的13对A亚组染色体上;各产生3对较强的GISH-NOR信号,其中1对分布在较长的A亚组上,2对分布在较短的D亚组上,其GISH-NOR信号强度与分布情况与以D基因组棉种为探针时相似.说明二倍体B、E、F基因组与四倍体棉A亚基因组具有较高的同源性,亲缘关系更近.这一点与它们的地理分布情况相符;而它们基因组中的45S rDNA重复序列与二倍体D基因组的45SrDNA重复序列同源性较高.C1和G1中以gDNA为探针时,杂交信号分布在2个栽培种全部26对染色体上,无法区分开A或D亚组染色体,都有3对较强的GISH-NOR信号.这一现象与D基因组拟似棉(D6) gDNA为探针的GISH相似,表明二倍体C和G基因组与四倍体棉的A和D亚基因组均具有较高的同源性,或者C和G基因组同时含有A基因组(或其他非洲棉基因组)和D基因组成分,进一步证实了其基因组成分的杂合性;而它们基因组中的45S rDNA重复序列同属D基因组类型.[结论]这些发现可为棉花杂交育种和棉属起源与演化研究提供有用信息.     

Selection of genetically diverse sea oats lines with improved performance for coastal restoration in the northern Gulf of Mexico

Numerous sea oats (Uniola paniculata L.) plants are transplanted to Northern Gulf of Mexico beaches each year to reduce coastal erosion. To adapt to environmental changes and effectively reduce coastal erosion, genetically diverse sea oats plants with demonstrated plant performance must be used. The objectives of this study were to: (i) identify improved sea oats lines; and (ii) determine the genetic diversity of improved sea oats lines. From 2003 to 2005, 2,000 sea oats lines were evaluated in unreplicated field trials at natural beach sites. In 2005, 75 sea oats lines were selected based upon phenotypic performance. The 75 selected lines and 3 plants of ‘Caminada’, the only commercially available sea oats line, were evaluated in replicated field trials in 2008 and 2010. In 2008, UP01LA-15 K-HB-3092, UP01LA-16S-GP-3138, UP01LA-31-GP-3103, UP01LA-33-GP-1303, and UP01NC-04-HB-3374 had higher (p < 0.05) stem densities than Caminada. In 2010, variation was not detected for any trait measured; lack of significant differences was most likely due to plant stress caused by storm surge one week after transplant. Amplified Fragment Length Polymorphism detected 534 loci for the 75 selected lines and 3 Caminada plants. One hundred eighty one loci were polymorphic; the average polymorphism rate was 34% (range = 25–43%). Polymorphic information content values ranged from 0.15 to 0.41 and averaged 0.29 while Jaccard similarity coefficients ranged from 0.8243 to 0.9794. These findings demonstrate the application of plant breeding techniques to develop genetically different sea oats lines with improved performance for coastal restoration projects in the northern Gulf of Mexico.     

禾本科植物自交不亲和基因定位进展

植物的自交不亲和性普遍存在于禾本科植物中,这是植物在长期进化过程中形成的防止自交退化,且有利于异花授粉、保持遗传多样性的一种生殖隔离机制。并且植物的自交不亲和机制多种多样,调控其性状的基因数量也不等。目前,许多实验已经证明禾本科植物的自交不亲和性属于S-Z型双因子自交不亲和系统,该类型的自交不亲和系统受到2个不连锁的复等位基因座S和Z的控制。本研究介绍了禾本科植物双因子S-Z型自交不亲和系统的起源,并且归纳了多年来对黑麦、天蓝虉草、球茎大麦和黑麦草等禾本植物自交不亲和调控基因的研究进展。还对禾本科植物S-Z双因子自交不亲和系统提出质疑。最终提出通过克服长雄野生稻的自交不亲和性来构建自交不亲和群体,从而用于精细的图位克隆。     

The germination of freshly harvested seed of ripe and unripe barley and oats

Barley and oats were grown in a greenhouse and outside in the field to different stages of ripeness. After harvest the plants were threshed as soon as possible. The seeds were treated in different ways and sown immediately in a sand-bed. The kernels which germinated were counted.Seeds from plants which were grown in the greenhouse, on average germinated better than seeds from plants grown outside. Seeds from ripe plants germinated better than green seeds, but in some trials germination of green seeds was exceptionally good. The percentage of germination was increased by several treatments. Drying at 50°C was found to be the most efficient method.The result from such experiments may be different when the plants are grown in another environment. Under our conditions drying at a temperature of about 50°C was an efficient way to increase the germination of freshly cut seeds of barley and oats.     

Molecular cloning of Tulipa fosteriana rDNA and subsequent FISH analysis yields cytogenetic organization of 5S rDNA and 45S rDNA in T. gesneriana and T. fosteriana

In this study, Tulipa fosteriana was found to contain 45S rDNA repeat units of 9.7 and 9.5 kb, in which at least 7 types of 45S rDNAs were identified by restriction site analysis. For 5S rDNA, repeat units ranging from 364 bp to 396 bp were identified. The diploid cultivars (2n = 2x = 24) ‘Christmas Dream’ and ‘Queen of Night,’ representing the horticultural group T. gesneriana, and ‘Red Emperor’, belonging to T. fosteriana, were compared cytogenetically using cloned 5S and 45S rDNAs. Fluorescence in situ hybridization (FISH) analysis identified many rDNA sites located on each chromosome in the diploid genomes. For example, we identified 71 sites of 5S rDNA and 10 sites of 45S rDNA in ‘Red Emperor’. Additionally, FISH analyses enabled construction of karyotypes for these cultivars. Karyotype comparison of T. gesneriana cultivars showed conservation of repetitive rDNA unit positioning. A clear difference in chromosome size and signal pattern was observed between T. gesneriana and T. fosteriana cultivars. Here we demonstrate the unique nature of the highly repeated 5S rDNA units in these Tulipa species and the usefulness of FISH karyotyping with cloned 5S and 45S rDNAs to clearly distinguish between chromosomes from T. gesneriana and T. fosteriana. Hitoshi Mizuochi and Agnieszka Marasek contributed equally to this paper     

Karyotype analysis and physical mapping of 45S rRNA genes in Hydrangea species by fluorescence in situ hybridization

Detailed karyotypes of Hydrangea macrophylla, Hydrangea paniculata and Hydrangea quercifolia were constructed on the basis of arm lengths and centromeric index, together with 45S rDNA fluorescence in situ hybridization. Although the chromosomes were small, they were well distinguishable for all species. Chromosome morphology and karyotypes were different for the three species. H. macrophylla had six metacentric (M), eight submetacentric (SM) and four subtelocentric (ST) chromosomes. The karyotype of H. paniculata contained seven M, 10 SM and one ST chromosomes and H. quercifolia had six M, 10 SM and two ST chromosomes. The variability among three species also was expressed by 45S rDNA signals. H. macrophylla had a nucleolar organizing region on chromosome 2, H. paniculata had 45S rDNA signals on chromosomes 2, 5 and 11 and H. quercifolia on chromosomes 3 and 8. Hybridization signal always was distally on the short arm but the strength of the signals was different for the three species. The chromosome portraits made in this study will be used to trace chromosome behaviour in interspecific hybrids resulting from breeding work between the three species.     

高寒区旱地莜麦合理种植密度的研究

通过对旱地莜麦不同种植密度的试验,进一步研究了密度对莜麦主要农艺状、干物质积累、叶面积系数及产量构成因素的影响,确定出在中等肥力条件下,晋北高寒区旱地莜麦的适宜种植密度为:播量600—675万粒/hm2,基本苗400—500万株/hm2。在该密度范围之内,群体和个体生长良好,产量各构成因素间关系协调,合理密植是获得旱地莜麦高产、稳产的关键技术。