Retrotransposon-based molecular markers for grapevine species and cultivars identification

Insertional polymorphisms of two copia-like (Vine-1, Tvv1) and one gypsy-like (Gret1) retrotransposon found in the grapevine genome were studied in 29 Vitis genotypes (Vitis arizonica, Vitis cinerea, Vitis labrusca, Vitis rupestis, Vitis rotundifolia, Vitis vinifera subsp. sylvestris and 23 V. vinifera subsp. sativa) using inter-retrotransposon amplified polymorphism (IRAP), retrotransposon-microsatellite amplified polymorphism (REMAP) and sequence-specific amplified polymorphism (SSAP) techniques. IRAP, REMAP and SSAP polymorphisms were compared with amplified fragment length polymorphism (AFLP), Inter-single sequence repeats (ISSR) and SSR polymorphisms by evaluating the information content, the number of loci simultaneously analysed per experiment, the effectiveness of the analyses in assessing the relationship between accessions and the number of loci needed to obtain a coefficient of variation of 10%. The UPGMA dendrograms of each molecular marker system were compared and the Mantel matrix correspondence test was applied. Furthermore, the corresponding insertion ages of the transposable elements were estimated for each retrotransposon subfamily analysed. The presence of Gret1, Tvv1 and Vine-1 retrotransposons in all analysed genotypes suggests that copia-like and gypsy-like retrotransposons are widespread in Vitis genus. The results indicate that these retrotransposons were active before Vitis speciation and contributed to Vitis genus evolution. IRAP, REMAP and SSAP markers allow the discrimination of Vitis species and V. vinifera subsp. sativa cultivars with certainty as has been shown with AFLP, ISSR and SSR analyses, but phylogenetic trees obtained by retrotransposon-based molecular markers polymorphisms show some significant differences in the allocation of the analysed accessions compare to those obtained by ISSR, AFLP and SSR molecular markers. The phylogenetic tree resulting from REMAP polymorphism appeared the most representative of the effective relationship between all analysed accessions.     

基于RAPD改进的分子标记技术

随机扩增多态DNA(RAPD)由于经济、快速简便、不需预知模板DNA信息、对模板DNA要求较低,因此被大量应用于昆虫学的研究中,如昆虫的鉴定分类、构建连锁图谱、系统发育分析等。但RAPD也有许多缺点,主要是一种显性标记而不能区分杂合体和纯合体,结果的重复性易受多因素的影响等。针对这些缺点,人们通过将RAPD与其它检测技术如聚丙烯酰胺凝胶电泳、单链构象多态(SSCP)、变性梯度凝胶电泳(DGGE)等相结合,提高了其分析的灵敏度和稳定性;再者,还将RAPD标记发展成为其它更稳定的或共显性的标记,如SCAR、微卫星等,为RAPD进一步的应用开阔了前景。文章对这些检测和转化的改进技术进行综述。     

沈阳城市森林主要绿化树种——丁香的光合特性研究

应用LI-6400红外气体分析仪对城市森林典型树种丁香不同季节光合作用的日变化规律进行研究,采用相关分析和多元逐步回归统计分析方法,得出净光合速率与光合有效辐射、气温、大气湿度、气孔导度等生理生态因子间的相关关系．结果表明,丁香春季和夏季光合速率呈双峰曲线,秋季呈单峰曲线．春季影响光合速率的主要因子为光合有效辐射和气孔导度;夏季影响光合速率的主要因子为气孔导度和光合有效辐射;秋季为气孔导度和胞间CO2浓度,建立了相应的影响因子回归方程。     

12份连翘种质资源的核型参数分析

采用常规压片法对4个连翘属原种以及6个金钟连翘品种和2个朝鲜连翘品种的核型进行了分析。研究结果发现,除金钟连翘品种Goldrausch为四倍体(2n=4x=56)外,其余11份种质均为二倍体(2n=2x=28),染色体基数为14。连翘、卵叶连翘、东北连翘、朝鲜连翘的核型分别为2A、3B、2B、3B型,6个金钟连翘品种的核型包括2A、2B、3B 3种,2个朝鲜连翘品种的核型包括2B、3B。12份连翘种质的着丝粒指数差异性不大,介于9.65%~50.00%,染色体类型以中部(m)、近中部(sm)以及近端部(st)染色体为主,未发现端部染色体和随体。结果表明,连翘和东北连翘的亲缘关系最近,与卵叶连翘的亲缘关系最远,金钟连翘品种与东北连翘、朝鲜连翘之间亲缘关系较近。研究结果为连翘属种间、品种间杂交育种和亲缘关系研究提供了细胞学基础。     

Cyanidin 3-p-coumaroylglucoside in Camellia species and cultivars

The major anthocyanin of red flowers of Camellia hiemalis, C. japonica and C. sasanqua was determined to be cyanidin 3-O-β-d-(6-O-p-coumaroylglucoside) by fast atom bombardment mass spectrometry and NMR spectroscopy. It was identical with the pigment, hyacinthin, of the bulb scales of Hyacinthus orientalis. This pigment and cyanidin 3-glucoside are widely distributed in the flowers of Camellia japonica and many Camellia cultivars.     

Screening of turfgrass species and cultivars for NaCl tolerance

Summary Information regarding the relative levels of salt tolerance between cultivars of Kentucky bluegrass (Poa pratensis L.) is lacking. The objectives of this study were to 1) develop a simple, quick and sensitive method of screening turfgrass species for NaCl tolerance and 2) to compare the relative salt tolerance of five cultivars of Kentucky bluegrass (Ram I, Adelphi, Baron, Bensun, and Nassau) to other known salt tolerant turfgrass species such as alkalaigrass (Puccinellia distans (L.) Parl. cv. Fults) and two cultivars of red fescue (Festuca rubra L. Dawson, and Checker).Alkalaigrass and both cultivars of red fescue retained a high level of salt tolerance compared to the Kentucky bluegrass cultivars. Significant variability in salt tolerance was apparent among the Kentucky bluegrass cultivars with Adelphi and Ram I exhibiting the best overall tolerance.     

Interspecific crosses between Tulipa gesneriana cultivars and wild Tulipa species: a survey

Summary Interspecific crosses were made between 76 Tulipa gesneriana cultivars and 28 botanical Tulipa species. Tulipa gesneriana proved fully congruent (all cross-combinations giving hybrid progeny) with members of the subsection Gesnerianae, was moderately affiliated (50% of all cross-combinations giving hybrid progeny) with representatives of the subsection Eichleres, and was found to be incongruent with most other Tulipa species (except T. stapfii of the subsection Oculussolis).Relationships between T. gesneriana and some representatives of the subsection Eichleres were addressed in more detail. Seeds were harvested in all cross-combinations, but the number of seeds depended on the T. gesneriana cultivar used as the female partner. Apparently, the maternal genotype did affect pre- or post-fertilization processes. T. gesneriana was not crossable with T. kaufmanniana of the subsection Spiranthera. In this specific case, however, a bridge cross was used to overcome incongruity. For this, T. kaufmanniana was first combined with T. greigii, and the hybrid was subsequently crossed with T. gesneriana.These results demonstrate possibilities for interspecific cross-combinations in tulips and have offered opportunities for the introduction of desired new traits in the cultivated tulip.     

Responses of various crop species and cultivars to fertilizer application

Summary Crop response to fertilizer application depends not only on the level of available plant nutrients in the soil but is also related to crop physiology and morphology. For a well balanced nutrition the rate of nutrient supply to the roots must correspond with the rate of nutrient required for growth. Species or cultivars with a high growth rate generally respond more favourably to fertilizer application than those with low growth rates. An analogous relationship holds for the biomass produced per unit soil surface. Thus modern rice and wheat cultivars tolerate a more dense spacing than older ones. Due to the dense stand the yield and particularly the grain yield of the modern varieties may be several times higher than those of older cultivars, and therefore also the nutrient requirement, especially the demand for N and P, is higher for the modern cultivars.Modern cereal cultivars are characterized by a high crop index which means that after flowering a high proportion of grain filling material must be produced by photosynthesis. Assimilation and translocation of photosynthates are favoured by K⁺. Thus in particular modern cultivars require a high K⁺ content for optimum grain filling.Nutrient exploitation of soils by plant roots depends on root morphology and root physiology. Grasses generally have much longer roots than dicots. Thus the rate of K⁺ and phosphate uptake per unit root length is lower for grasses than for dicots. It is for this reason that dicots respond earlier to a K⁺ and phosphate dressing than grasses.Species living symbiotically with Rhizobium may depress the rhizosphere pH considerably and thus promote the dissolution of phosphate rock.     

Molecular species of glycinin in some soybean cultivars

A(4) polypeptide-containing (Shirotsurunoko and York) and A(4) polypeptide-lacking (Raiden and Suzuyutaka) soybean cultivars were used to investigate the heterogeneity of glycinin molecular species. Purification of glycinin by DEAE-Toyopearl column chromatography afforded molecular species eluting before the glycinin fraction. Analysis of this fraction by SDS-polyacrylamide gel electrophoresis (SDS-PAGE) and sucrose density gradient centrifugation indicated that this protein consisted of A(1) and A(2) polypeptides. The A(4)-containing soybean cultivars contained less of this protein than the A(4)-lacking soybean cultivars, as exhibited by the size of the early peak appearing during column chromatography. Alkaline PAGE and N-terminal amino acid sequence analysis confirmed that the A(1)- and A(2)-rich molecular species in the A(4) polypeptide-lacking cultivars consisted of the A(1a) and A(2) polypeptides. Estimation of the molecular mass by gel permeation chromatography and multi-angle laser light scattering (GPC-MALLS) indicated that the A(1a)- and A(2)-rich molecular species were similar to a monomer of glycinin.     

Testing acoustic versus physical marking: two complementary methods for individual-based monitoring of elusive species

Individuals of some species differentiate each other on the basis of the acoustic features of their vocalizations, and this can be used in individual‐based population monitoring studies. No research has tested for the effectiveness of individual marking through voice recognition as compared to traditional monitoring methods relying on physical marks. We compared voice recognition and physical marking using the Dupont's lark Chersophilus duponti as a study species. This bird needs to be attracted with playback in order to be seen (or captured). We first demonstrated that the territorial calls from a sample of banded males were individually distinctive and constant over time by means of discriminant function analysis, which correctly classified 100% of marked males. Then, we applied similarity techniques on call spectrotemporal features to define a threshold value of similarity within banded individuals, to be combined with qualitative spectrogram inspection for the classification of all recorded birds. Eventually, we compared the voice and the capture samples, to test for differences in relation to re‐location rate, territory fidelity and dispersal movements both within and between years. Voice recognition was less time‐consuming than capture‐recapture method in the field, but it was useless for monitoring yearlings in call development stage. The two methods provided the same results in terms of territory fidelity and dispersal movements, but differed in re‐location rates, which were significantly greater in the case of voice recognition method. By means of physical captures we possibly trapped a large sample of young and silent floaters, with low probability of recapture or recording. This mismatch between methods could bias the estimates of annual survival, which strongly depend on re‐location rates. We suggest considering the two methods as complementary rather than alternatives for monitoring populations. Each technique offers unique information, and the two sources should be combined to provide correction factors that would eventually sharpen our knowledge on bird population ecology.     

The complete chloroplast genome sequence of the endangered species Syringa pinnatifolia (Oleaceae)

Syringa pinnatifolia is an endangered endemic species in China with important ornamental and medicinal value, and it needs urgent protection. Here, we report the complete chloroplast (cp) genome structure of S. pinnatifolia and its evolution is inferred through comparative studies with related species. The S. pinnatifolia cp genome was 155 326 bp and contained a large single copy region (LSC) of 86 167 bp and a small single copy region (SSC) of 17 775 bp, as well as a pair of inverted repeat regions (IRs) of 25 692 bp. A total of 113 unique genes were annotated, including 79 protein‐coding genes, 30 tRNA genes and four rRNA genes. The GC content of the S. pinnatifolia cp genome was 37.9%, and the corresponding values in the LSC, SSC and IR regions were 36.0, 32.1, 43.2% respectively. Repetitive sequences analysis revealed that the S. pinnatifolia cp genome contained 38 repeats. Microsatellite marker detection analysis identified 253 simple sequence repeats (SSRs), which provides opportunities for future studies of the population genetics and phylogenetic relationships of Syringa. Phylogenetic analysis of 29 selected cp genomes revealed that S. pinnatifolia is closely related to Syringa vulgaris and all 27 Lamiales species formed a clade separate from the two outgroup species. This newly characterized S. pinnatifolia chloroplast genome will provide a useful genomic resource of phylogenetic inference and the development of more genetic markers for species discrimination and population studies in the genus Syringa.     

Genetic diversity of ornamentalAllium species and cultivars assessed with isozymes

The aim of the study was to verify the similarity of 13 species and 5 cultivars of ornamental alliums and classify them into groups based on morphological and isozyme variation. The work embraced: Allium aflatunense, A. caeruleum, A. christophii, A. giganteum, A. karataviense, A. moly, A. nigrum, A. pyrenaicum, A. rosenbachianum, A. schubertii, A. siculum (syn. Nectaroscordum siculum), A. sphaerocephalon, A. strictum, A. stipitatum 'Album', A. 'Ivory Queen', A. 'Lucy Ball', A. 'Mont Blanc', and A. 'Purple Sensation'. Scape length, inflorescence diameter, and flowering period were recorded. Isozyme marker polymorphism was assessed by starch gel electrophoresis. Eight polymorphic isozyme systems (AAT, GPI, PGM, ALAT, ACP, DIAP, ALDO, PGD) were selected from 16 analysed in the taxa. Besides the differences between the taxa, the isozymes revealed intraspecific polymorphism in 5 systems. A total of 37 markers were obtained and used for dendrogram construction. The most similar taxa were A. karataviense with A. 'Ivory Queen', and A. karataviense with A. christophii (similarity level 0.78). A high similarity of 11 taxa belonging to one group (A. aflatunense, A. christophii, A. giganteum, A. karataviense, A. nigrum, A. schubertii, A. 'Ivory Queen', A. 'Lucy Ball', A. stipitatum 'Album', A. 'Mont Blanc', A. 'Purple Sensation') suggested that this group could be identified with the subgenus Melanocrommyum.     

Cardiolipin: characterization of distinct oxidized molecular species

Cardiolipin (CL) is a phospholipid predominantly found in the mitochondrial inner membrane and is associated structurally with individual complexes of the electron transport chain (ETC). Because the ETC is the major mitochondrial reactive oxygen species (ROS)-generating site, the proximity to the ETC and bisallylic methylenes of the PUFA chains of CL make it a likely target of ROS in the mitochondrial inner membrane. Oxidized cellular CL products, uniquely derived from ROS-induced autoxidation, could serve as biomarkers for the presence of the ROS and could help in the understanding of the mechanism of oxidative stress. Because major CL species have four unsaturated acyl chains, whereas other phospholipids usually have only one in the sn-2 position, characterization of oxidized CL is highly challenging. In the current study, we exposed CL, under aerobic conditions, to singlet oxygen (¹O₂), the radical initiator 2,2′-azobis(2-methylpropionamidine) dihydrochloride, or room air, and the oxidized CL species were characterized by HPLC-tandem mass spectrometry (MS/MS). Our reverse-phase ion-pair HPLC-MS/MS method can characterize the major and minor oxidized CL species by detecting distinctive fragment ions associated with specific oxidized species. The HPLC-MS/MS results show that monohydroperoxides and bis monohydroperoxides were generated under all three conditions. However, significant amounts of CL dihydroperoxides were produced only by ¹O₂-mediated oxidation. These products were barely detectable from radical oxidation either in a liposome bilayer or in thin film. These observations are only possible due to the chromatographic separation of the different oxidized species.     

Meiosis in some species and cultivars ofCrocus (Iridaceae)

Meiosis is described in thirty-seven taxa ranging from low to high chromosome numbers. Most were regular and bivalent forming.C. flavus subsp.flavus was observed at first anaphase only and was highly irregular.C. speciosus had one or two possible interchange quadrivalents and theC. vernus aneuploid cultivars exhibited multivalents, bivalents and univalents together with some anaphase bridges and fragments. Chiasma frequency is determined here for the first time.     

The Genus Syringa: Molecular Markers of Species and Cultivars

RAPD analysis was carried out with 22 accessions of the genus Syringa, including six species, one interspecific hybrid, and 15 cultivars. In total, 512 polymorphic fragments were detected; species-specific and cultivar-specific markers were identified. For the first time, genetic polymorphism and genome similarity coefficients were estimated and phylogenetic relationships were established for the genus Syringa.     

Plant regeneration from Citrus protoplasts: Variability in methodological requirements among cultivars and species

Summary Nucellar callus lines were established from two orange cultivars (Nucellar Shamouti, Shamouti Landau), three mandarin cultivars (Murcott, Dancy, Ponkan) one grapefruit cultivar (Duncan) and sour orange (Citrus aurantium). These callus lines were initiated from in vitro cultured ovules of young fruits and maintained an embryogenic capacity. The plating efficiencies of protoplasts derived from these calli, as well as those of protoplasts from lemon (cv. Villafranca) nucellar callus were differentially affected by the maceration enzymes and by the sugars used as osmotic stabilizers. Plants with normal morphological features were regenerated from cultured protoplasts derived from each of the nucellar callus lines. The establishment of eight new protoplast systems in Citrus paves the way for cell genetics studies and for novel breeding approaches in these economically important orchard trees.Contribution from the Agricultural Research Organization, The Volcani Center, Bet Dagan, Israel. No. E-290, 1981 series     

Genetic relationships of Aglaonema species and cultivars inferred from AFLP markers

BACKGROUND AND AIMS: Aglaonema is an important ornamental foliage plant genus, but genetic relationships among its species and cultivars have not been reported. This study analysed genetic relatedness of 54 cultivars derived from nine species using amplified fragment length polymorphism (AFLP) markers. METHODS: Initially, 48 EcoRI + 2/MseI + 3 primer set combinations were screened, from which six primer sets that showed clear scoreable and highly polymorphic fragments were selected and used for AFLP reactions. AFLP fragments were scored and entered into a binary data matrix as discrete variables. Jaccard's coefficient of similarity was calculated for all pair-wise comparisons among the 54 cultivars, and a dendrogram was constructed by the unweighted pair-group method using the arithmetic average (UPGMA). KEY RESULTS: The number of AFLP fragments generated per primer set ranged from 59 to 112 with fragment sizes varying from 50 to 565 bp. A total of 449 AFLP fragments was detected, of which 314 were polymorphic (70 %). All cultivars were clearly differentiated by their AFLP fingerprints. The 54 cultivars were divided into seven clusters; cultivars within each cluster generally share similar morphological characteristics. Cluster I contains 35 cultivars, most of them are interspecific hybrids developed mainly from A. commutatum, A. crispum or A. nitidum. However, Jaccard's similarity coefficients among these hybrids are 0.84 or higher, suggesting that these popular hybrid cultivars are genetically much closer than previously thought. This genetic similarity may imply that A. nitidum and A. crispum are likely progenitors of A. commutatum. CONCLUSIONS: Results of this study demonstrate the efficiency and ease of using AFLP markers for investigating genetic relationships of ornamental foliage plants, a group usually propagated vegetatively. The AFLP markers developed will help future Aglaonema cultivar identification, germplasm conservation and new cultivar development.