基于SSR分子标记分析云南月季种质资源亲缘关系(英文)

利用简单重复序列SSR(simple sequence repeat)标记技术对48份月季种质资源(包括14份野生种、20份古老月季品种和14份栽培品种)的遗传多样性和亲缘关系进行了分析.结果表明,(1)18对SSR引物在18个SSR位点上共检测到160个等位基因,每一位点的等位基因变幅为6～13个,平均8.9个,材料间遗传相似系数变化范围为0.157 8～0.754 9,表明在分子水平上云南月季种质资源具有丰富的遗传多样性.(2)聚类分析结果显示,在相似系数为0.44处,可将 14个野生种明显分为6个组,这与植物形态学分类结果上基本一致;在遗传相似系数为0.40水平上将48份供试材料分为八大组群.(3)亲缘关系分析结果显示,5个野生种和所有古老月季品种与大多数栽培品种的亲缘关系较近.     

月季种质资源的倍性变异及核型多样性研究(英文)

采用常规压片法对包括5个蔷薇野生种、8个古老月季及9个现代月季品种共22份月季种质资源进行核型分析。供试材料共有3种不同倍性:5个野生种中粉团蔷薇为三倍体(2n=3x=21),其他4个材料为二倍体(2n=2x=14);9个现代月季品种均为四倍体(2n=4x=28);而古老月季品种中包含二、三、四倍体3种倍性。22份材料中共发现4种不同类型的核型,在野生种中为1A、2A;古老月季中为1A、2A、1B;现代月季中为1A、1B、2B。另外,供试材料在核不对称系数、着丝粒指数、随体数目及位置、染色体相对长度组成等方面差异较大,说明在月季种质资源中存在着丰富的核型多样性。     

小麦低分子量谷蛋白亚基基因5′侧翼保守序列染色体组特异性DNA变异的分析及验证

根据33个低分子量麦谷蛋白亚基(LMW-GS)基因5′侧翼序列的相似性进行聚类分析, 可将其划分成8个类群, 这与基于N末端推导氨基酸序列进行的类群划分结果完全一致. 序列比对发现, 各类群基因5′侧翼保守序列间存在DNA多态性, 共发现34个多态性位点, 其中18个为潜在单核苷酸多态性位点(SNPs, single nucleotide polymorphisms). 除1个LMW-GS类群之外, 其余7个类群的5′侧翼序列均具有类群特异性DNA变异位点. 根据类群间的DNA多态性对这7个类群设计了特异引物, 利用普通小麦(Triticum aestivum L.)品种中国春及其第1同源群双端体系对其进行染色体定位分析, 揭示了1AS, 1BS和1DS上分别有第2, 1和4类群. 对PCR产物的克隆测序进一步验证了不同染色体组上的LMW-GS基因类群间5′侧翼序列具有特异性. 这些结果表明, LMW-GS基因的编码区及其5′侧翼保守序列可能是协调进化的. 本文报道的7对引物可对7类LMW-GS基因的完整编码区进行特异扩增, 因而能在小麦复杂的遗传背景下有目的地对某一类LMW-GS基因进行分离克隆, 这有助于弄清单个LMW-GS对小麦品质的贡献. 同时, 在小麦育种中, 这些标记对于有效地选择与品质密切相关的LMW-GS组分有一定应用价值.     

Uncv无毛小鼠无毛基因的分子克隆及序列分析

目的 探讨Unev无毛小鼠的无毛性状与无毛基因(hairless gene,hr)的相关性.方法 参照Gen-Bank上公布的小鼠的hr序列,设计5对引物,用RT-PCR方法对本单位培育的Unev无毛小鼠hr的编码区序列进行了克隆与分析.结果 获得了Unev无毛小鼠及野生型hr的全部编码区序列(3546 bp).Unev无毛小鼠hr基因与野生型小鼠hr基因的长度及序列完全一致,同源性为100%.与GenBank上发表的国外小鼠hr基因序列(Z32675)相比,同源性为99.7%,共10个碱基发生了突变,其中2个碱基突变导致了相应的氨基酸突变;和昆明小鼠的hr(AY547391)相比,同源性为99.6%,共12个碱基发生了突变,其中3个碱基突变导致了相应的氨基酸突变;但这些突变是由种属差异造成的.结论 Uncv无毛小鼠的无毛性状产生与hr基因无关.     

部分月季花品种的数量分类研究

用数量分析的方法,对80个月季、蔷薇及其远缘杂种的样本进行了系统分析与整理。Q型聚类分析结果表明,可将80个样本分为9类,并初步揭示了各类型间的亲缘关系。首次提出了将中国古老月季分为高大和低矮两个型,将新培育出的具有较高抗性的远缘杂种月季另立为灌丛月季型的分类方案。同时,分析并预测了采用中国原产的蔷薇野生种与古老月季品种或现代月季品种杂交,后代可能形成的月季品种类型。并对花卉品种数量分类中的一些方法进行了初步探讨。     

中国古老月季‘月月粉’FT/TFL1基因的鉴定与表达分析

该研究以中国古老月季品种‘月月粉’(Rosa chinensis‘Old Blush’)为试材,根据拟南芥和草莓FT/TFL1基因家族成员的保守结构域设计引物,对‘月月粉’基因组DNA进行克隆和测序,结合数据库序列和双单倍体基因组信息进行序列比对,结果共获得了8条月季的FT/TFL1基因家族序列,且8条序列分别注释为2个FT基因(RcFT1和RcFT2)、1个BFT基因(RcBFT)、1个ATC基因(RcATC)、2个MFT基因(RcMFT1和RcMFT2)和2个PEBP基因(PEBP1和PEBP2);进化树分析结果将这8个基因分为3个亚组;结构分析显示8个基因的内含子为1～5个不等。qRT-PCR分析显示,RcFT1、RcPEBP、RcMFT1、RcMFT2和RcBFT在‘月月粉’的叶片中表达量最高,茎尖中次之;RcFT2在茎尖中的表达最高,叶片中次之;RcATC在茎尖中的表达量高于其他组织;除RcPEBP在根组织中有表达外,其他基因在根组织中几乎没有检测到。进一步表达分析显示,RcFT1和RcFT2在‘无刺光叶蔷薇’(R.wichuriana‘Basye’s Thornless')生殖生长期的叶片、茎尖中的表达量显著高于营养生长期。研究推测,具有连续开花性状的中国古老月季‘月月粉’可能具有多个开花时间(FT)基因位点,FT有可能可以作为月季生殖转换的标记基因。     

七里香蔷薇叶绿体基因组测序及结构分析

七里香蔷薇是蔷薇属木香花的一个变种,在秦岭山区广泛分布,具有重要的观赏价值,同时也是蔷薇属花卉育种的重要种质资源。本研究首次以秦巴山区七里香蔷薇为实验材料,采用Illumina HiSeq4000测序系统对其叶绿体基因组DNA建库测序,建库类型为350 bp DNA小片段文库,测序深度为59×。分析其基因组结构,并且与已报道蔷薇科植物叶绿体基因组进行比较。七里香蔷薇叶绿体基因组序列全长156 544 bp,具有典型被子植物叶绿体基因组环状四分体结构。叶绿体基因组包括大单拷贝区85 678 bp、小单拷贝区18 782 bp和2个反向重复序列26 042 bp。GC含量为37.22%;检测到116个串联重复序列43个散在重复序列;此外,还检测到72个简单重复序列位点。对其功能基因比对分析,该植物的叶绿体基因组共注释得到129个基因,包括86蛋白编码基因、35个tRNA基因和8个rRNA基因。用MEGA6.0软件通过邻近法基于叶绿体全基因组序列对所有已知叶绿体基因组的蔷薇科22个物种进行聚类分析,其与七里香蔷薇和大花香水月季、缫丝花聚在一起。研究结果对于七里香蔷薇野生植物资源的育种及引种驯化具有参考依据。     

枯草芽孢杆菌抗脯氨酸结构类似物突变株的筛选及突变株proBA基因的克隆和序列分析

利用亚硝基胍对枯草芽孢杆菌93151进行诱变处理,获得了耐NaCl浓度达14％的突变株,同时发现该突变株也是一个抗脯氨酸反馈抑制突变菌株,其胞内自由脯氨酸的含量随着盐浓度的提高显著增加,说明其对渗透压的耐受能力与胞内自由脯氨酸的含量紧密相关。利用PCR方法克隆突变株的proBA基因,得到一个约2.3kb的DNA片段,序列分析表明该片段含有一完整的proB基因和部分proA基因,与野生菌株的proB基因相比,突变株proB基因中有3个碱基发生了改变,其中一个碱基的变化（从起始密码子开始第781位由T→A）导致了一个氨基酸发生改变（Ser→Thr),另外两个碱基变化为沉默位点突变。将该proB基因转入大肠杆菌脯氨酸营养缺陷型菌株,能够与其功能互补。同时对部分proA基因序列分析发现,其与proB基因头尾重叠。在proA基因起始密码子上游第14个碱基处有一个类似于SD的序列,其所编码的氨基酸序列与枯草芽孢杆菌168的同源性为77％。     

黑腹果蝇黑条体突变体突变位点的鉴定

根据果蝇ebony基因的序列设计了一系列特异性引物,对1991年发现的一种黑腹果蝇的新的体色突变（Qian＆Zhang,1994）——黑条体突变型的ebony基因进行克隆和序列测定。与野生型W91910的ebony基因相比,黑条体突变型的ebony基因的编码区无明显的大片段变异,变异仅存在于数个氨基酸位点,且均不位于该基因产物的关键序列。在黑条体的ebony基因的5’端序列的克隆和序列测定中发现,与野生型w^91910及黑檀体e。的ebony基因相比,黑条体突变型的ebony基因有一个大片段的缺失,该缺失包括外显子1的206个碱基和内含子1的747个碱基,由此确定了黑条体突变体的突变类型和突变位点。     

林麝MHC-DRA基因分析及与反刍亚目DRA基因比较

MHC-DRA基因仅在少部分物种如马、斑马、驴等具有多态性,DRA基因在其他哺乳动物中多为低多态性。由于MHC-DRA的低多态性,较少受到研究者重视,我们选取近缘物种牛的DRA基因引物,对四川养麝研究所的217只林麝进行DRA基因exon 2的序列扩增,得到2个林麝DRA基因,片段长度为260 bp,这2个基因仅有一个核苷酸同义突变,没有氨基酸突变。我们将所获得的林麝DRA基因与反刍亚目其他物种DRA基因进行氨基酸比对发现,我们扩增的序列为林麝DRA基因exon 2,编码MHCⅡ类分子的α1区域,而这段区域主要参与形成MHCⅡ类分子的多肽结合槽,根据Brown确定人类的抗原肽结合区位点,在参与比对的反刍亚目物种DRA基因中,抗原位点11、22、72、76存在氨基酸变异,而在这些位点上林麝的DRA基因没有变异,和大部分反刍亚目相同。同时我们发现大部分反刍亚目物种之间氨基酸突变的位点大都在非抗原结合位点。     

Anthocyanins in flowers of genus Rosa, sections Cinnamomeae (=Rosa), Chinenses, Gallicanae and some modern garden roses

Forty-four taxa of three sections (Cinnamomeae (=Rosa) 26, Chinenses 8 and Gallicanae 10) and eight modern garden roses in the genus Rosa were surveyed for their floral anthocyanins. Eleven anthocyanins: 3-glucosides and 3,5-diglucosides of cyanidin (Cy), pelargonidin (Pg) and peonidin (Pn), 3-rutinosides and 3-rho-coumaroylglucoside-5-glucosides of Cy and Pn, and Cy 3-sophoroside, were isolated from flowers of these taxa and identified by chemical and spectroscopic techniques. Four anthocyanins: Cy 3-rutinoside, Pn 3-rutinoside, Pn 3-rho-coumaroylglucoside-5-glucoside and Cy 3-sophoroside were found for the first time in Rosa flowers.Investigated sections of wild roses showed characteristic distribution of anthocyanins. Cy 3,5-diglucoside was the dominant anthocyanin detected in all three sections, but accumulation of Pn 3,5-diglucoside distinguished sections Cinnamomeae from other sections, and the occurrence of Cy 3-glucoside separates section Chinenses from others.Cy 3-sophoroside was detected in large amount in some taxa of section Cinnamomeae: e.g., R. moyesii and its related cultivars, and R. rugosa cv. Salmon Pink. The acylated Cy glycoside was found in all sections and also in some modern garden roses, while the acylated Pn glycoside was detected in the section Cinnamomeae, but not in sections Chinenses and Gallicanae. According to anthocyanin distribution patterns, eight groups were classified chemotaxonomically in genus Rosa.     

应用流式细胞术测定17种中国野生蔷薇核DNA含量

以17种中国野生蔷薇为试材,采用改良的LB 01裂解液,以4种不同的标准植物——大豆（Glycine max Merr.‘Polanka’）、绿豆（Vigna radiata（L.） Wilczek）、番茄（Lycopersicon esculentum Miller）和玉米（Zea mays L.）为外标,以二倍体材料丽江蔷薇（Rosa×lichiangensis Yü et Ku）为内部参照,利用流式细胞术对其核DNA含量及染色体倍性进行检测,并采用常规染色体压片法验证倍性准确性。本研究首次检测了3个二倍体种——商城蔷薇（Rosa shangchengensis T.C.Ku）、广东蔷薇（Rosa kwangtungensis Yü et Tsai）和无刺刺梨（Rosa roxburghii f.inermis S.D.Shi）,1个三倍体种——伞房蔷薇（Rosa corymbulosa Rolfe）和1个四倍体种——弯刺蔷薇（Rosa beggeriana Schrenk）的核DNA含量及基因组大小。结果表明,流式细胞术检测结果与常规染色体压片法结果一致,可对中国野生蔷薇的倍性研究进行补充。本研究结果可丰富中国蔷薇属植物的细胞遗传学背景资料并为繁育新品种提供理论依据。     

Conservation measures for <Emphasis Type="Italic">Rosa arvensis</Emphasis> Huds. in Flanders (Belgium) based on congruent genetic and phenotypic population differentiation

Rosa arvensis is a naturally rare and scattered indigenous wild rose species in Flanders, the northern part of Belgium. As is the case for many light demanding woody species in this area, it is currently threatened by habitat fragmentation and destruction due to high human pressure. Recent inventories revealed a restricted distribution pattern for this rose, concentrated mainly in two regions of the south western part of Flanders. Surprisingly, strong differentiation was observed among natural populations in these two proximate regions in both an AFLP-based and a morphological analysis. A common garden experiment indicated a partly genetic basis for the morphological divergence. Additionally, the AFLP analysis of roses sampled in the same forested area within one of the two regions resulted in two differentiated gene pools. Possible causes for the observed differentiation can be adaptive divergence, founder effects and/or historical hybridisation with dogroses. Together, the congruent genetic and morphometric differentiation between the two geographic regions urges a cautious approach in conservation programs.     

基于ITS和matK基因对牡丹组序列分析及其亲缘关系的研究

以芍药属牡丹组全部9个野生种、5个紫斑牡丹栽培品种及3个中原牡丹品种为试材,进行核糖体内转录间隔区（ITS）和叶绿体成熟酶K （matK）基因片段测序分析,探讨ITS和matK序列为牡丹组所有野生种种间关系提供分子证据。从GeneBank中选取了1个牡丹及3个外类群芍药、川赤芍和草芍药的ITS及matK序列。对试验样品进行DNA提取、PCR扩增并双向测序得到44条序列,人工校正后将所得44条序列进行比对;计算碱基组成频率、变异位点、简约信息位点数、转换/颠换比率、种内及种间遗传距离,以邻接法进行系统发育分析。结果表明,牡丹组所有个体ITS序列长度在750~800 bp,含有86个多态位点,74个简约信息位点,转换/颠换比率（R）为1.2;而matK序列含有20个简约信息位点,转换/颠换比率（R）为1.7。ITS序列分析将牡丹组野生种分为两大枝,稷山牡丹、紫斑牡丹、卵叶牡丹和杨山牡丹聚为一枝,狭叶牡丹、滇牡丹、黄牡丹和大花黄牡丹聚为另一枝,这两枝分别与革质花盘亚组和肉质花盘亚组相对应,而四川牡丹位于革质花盘亚组最底端,支持前人研究将四川牡丹归为革质花盘亚组。matK序列分析的牡丹组野生种间遗传距离结果不理想,未能清晰的表明野生种之间的亲缘关系。由此说明,ITS序列更适合牡丹组野生种间亲缘关系的研究分析。     

Physical mapping of ribosomal DNA on several species of the subgenus Rosa

The localization of NORs by fluorescent in situ hybridization on metaphase spreads of five diploid (Rosa gigantea Coll., Rosa moschata Herrm., Rosa multiflora Thunb., Rosa rugosa Thunb. and Rosa sempervirens L., 2n=2x=14), one triploid (Rosa chinensis’semperflorens’ Koehne., 2n=3x=21) and one tetraploid (Rosa gallica ’versicolor’ L., 2n=4x=28) ancestral species of modern roses was studied. Two terminal hybridization signals were observed in all diploid species corresponding to a single NOR per genome in these species. Triploid R. chinensis showed three hybridization sites on the short arm of three morphologically similar chromosomes. Six hybridization sites, located at terminal positions of the short arms of three chromosome pairs, were observed in the tetraploid species. These signals corresponded to three pairs of NORs and all of them were located in chromosome pairs of different size. These observations, together with the analysis of meiotic pairing in PMCs, support the view that our specimen of R. chinensis is an autotriploid and that R. gallica’versicolor’ has an alloploidy nature. Received: 27 November 2000 / Accepted: 12 March 2001     

Triparental origin of Damask roses

Damask roses are one group of old rose varieties and a key material in old European rose improvement in the 19th century. To clarify the origin of Damask roses, we selected four varieties as the oldest Damask varieties and examined the relationship between the Damask varieties and their putative ancestors at the molecular level. Randomly amplified polymorphic DNA analysis of the Damask varieties proved that they had an identical profile, indicating they were established from a common ancestor. They have never been allowed to reproduce sexually; their reproduction depends entirely on vegetative propagation. We identified three Rosa species, R. moschata, R. gallica and R. fedschenkoana, as parental species of the original hybridization that contributed to forming the four oldest Damask varieties by sequencing the internal transcribed spacer of ribosomal DNA. We also found that all the four oldest Damask varieties had chloroplasts derived only from R. moschata, as judged from psbA-trnH spacer sequences. This triparental origin of the four oldest Damask varieties can explain some morphological characteristics of the four oldest Damask varieties, like fruit shape, leaf color and the 'Moss' character.     

Chemical and histochemical analysis of 'Quatre Saisons Blanc Mousseux', a Moss Rose of the Rosa x damascena group

BACKGROUND AND AIMS: Moss roses are old garden roses covered with a mossy growth on flower pedicel and calyx. This moss releases a pine-scented oleoresin that is very sticky and odoriferous. Rosa x centifolia 'muscosa' was the first moss rose to be obtained by bud-mutation but, interestingly, R. x damascena 'Quatre Saisons Blanc Mousseux' was the first repeat-blooming cultivar, thus interesting breeders. In the present study, the anatomy of these sports (i.e. bud-mutations) is characterized and the volatile organic compounds (VOCs) produced by the moss versus the petals are identified. They are compared between the two lines and their respective parents. METHODS: Anatomy of the moss is studied by environmental scanning electron microscopy and histochemical light microscopy. Sudan Red IV and Fluorol Yellow 088 are used to detect lipids, and 1-naphthol reaction with N,N-dimethyl-p-phenylenediamine to detect terpenes (Nadi reaction). Head-space or solid/liquid extraction followed by gas chromatography and mass spectrometry are used to identify VOCs in moss, trichomes and petals. KEY RESULTS: Moss of the two cultivars has the same structure with trichomes on other trichomes but not exactly the same VOCs. These VOCs are specific to the moss, with lots of terpenes. An identical VOC composition is found in leaves but not in petals. They are nearly the same in the moss mutants and in the respective wild types. CONCLUSIONS: Sepals of moss roses and their parents have a specific VOC pattern, different from that of the petals. The moss corresponds to a heterochronic mutation with trichomes developing on other trichomes. Such a mutation has probably appeared twice and independently in the two lines.     

Genetic differentiation between populations of the European rose hip fly Rhagoletis alternata

We tested for genetic differentiation between populations of Rhagoletis alternata Fall. (Diptera: Tephritidae) on three different host species. We collected larvae from three rose species of the section Caninae ( Rosa canina L., Rosa corymbifera Borkh . , and Rosa rubiginosa L.) from 15 sites across Germany, where the three roses occurred together. Additionally, we sampled three sites in Switzerland. Roses differ in morphology (e.g. leaf glands) as well as phenology. We were able to score nine allozyme loci (five polymorphic). Populations from the three hosts did not differ in genetic variability. We found significant genetic differentiation between populations from different host species. However, the differentiation was very low (0.9%). Hence, we found no indication for host races. Furthermore, surprisingly little geographical structure of genetic differentiation was found between populations of this fruit fly across central Europe. We offer three mutually non-exclusive explanations for these findings. First, gene flow between populations of Rh. alternata is high. Second, the pattern of genetic differentiation is based on a recent expansion of the distributional range . Third, the ongoing gene flow between roses of the section Caninae acts as a hybrid bridge.  © 2007 The Linnean Society of London, Biological Journal of the Linnean Society , 2007, 90 , 619–625.