RAPD and RFLP markers tightly linked to the locus controlling carnation (Dianthus caryophyllus) flower type

 Flower doubleness as a breeding characteristic is of major importance in carnation (Dianthus caryophyllus), one of the major cut-flowers sold worldwide, since flower architecture is of the utmost value in ornamentals. Based on the number of petals per flower, carnations are grouped into “single”, “semi-double” and “double” flower types. The first have five petals and are easily distinguishable, but of no economic value to the carnation industry. Flowers of standard and spray varieties, which constitute the largest market share, are usually of the double and semi-double type, respectively. These flower types are not easily distinguishable due to phenotypic overlaps caused by environmental conditions. To study the inheritance of this trait, several progeny segregating for flower type were prepared. Based on the number of single-flower type fullsibs among the offspring, we found that this phenotype is expressed only in plants homozygous for the recessive allele and that a dominant mutation in this allele causes an increase in petal number. Using random decamer primers, we identified a random amplified polymorphic DNA (RAPD) marker which is tightly linked to this recessive allele. The RAPD marker was cloned and used to generate a restriction fragment length polymorphic (RFLP) marker. This RFLP marker could discriminate with 100% accuracy between the semi-double and double- flower phenotypes in carnations of both Mediterranean and American groups. The advantages of RFLP over RAPD markers and their applicability to markerassisted selection in carnation are discussed. Received: 11 August 1997 / Accepted: 22 August 1997     

Contribution to the taxonomy and nomenclature of the vascular plants of Morocco

ROMO, A. M., 1992. Contribution to the taxonomy and nomenclature of the vascular plants of Morocco. As a result of studies on the flora of Morocco, a number of new taxa and a series of new nomenclature combinations are proposed which are more in line with current knowledge. Three new taxa are described: Dianthus atlantica from the Great Atlas; Ranunculus spicatus Desf. subsp. fontqueri Romo, endemic to Er Rif, and a hybrid: Saxifraga × fontqueri Romo (= S. crenata Psau & Font Quer × Saxifraga globulifera Desf.). The following new combinations are made: Astragalus gombo Cosson & Durieu subsp. pseudogombo (Fernandez Casas) Romo, Avenula bromoides (Gouan) H. Scholz subsp. cincinnatus (Ten.) Romo, Chaenorrhinum origanifolium (L.) Fourr. subsp. flexuosum (Desf.) Romo, Digitalis laciniata Lindley subsp. riphaea (Pau & Font Quer) Romo, Digitalis purpurea subsp. mauretanica (Humbert & Maire) Romo, Lavandula pedunculata Miller subsp. atlantica (Braun-Blanquet) Romo, Paeonia coriacea Boiss. var. maroccana (Pau & Font Quer) Romo, Sideritis arborescens Bentham subsp. antiallantica (Maire) Romo.     

Efficient transformation and regeneration of carnation cultivars usingAgrobacterium

We have developed an efficient method for transformation and regeneration of plants from carnation,Dianthus caryophyllus L. Whole leaves fromin vitro shoot cultures were mixed withAgrobacterium, cocultivated for 5 days and then plated on 2 µg/l chlorsulfuron (CS). Regenerated shoots and shoot clusters were divided into smaller sections and plated on 3 µg/l CS for selection to produce fully transformed shoots. Geneticin (G418) and kanamycin used were not as effective selective agents as CS. All regenerated shoots were vitrified. These were normalized, rooted and transferred to the greenhouse. 100% of regenerated plants were transformed based on rooting assay, GUS assay, PCR and Southern analysis.     

杂交石竹授粉受精及其胚胎发育过程的观察

以‘杂交石竹’为试验材料,利用荧光显微镜观察其授粉后花粉萌发、花粉管生长情况,采用石蜡切片法对其受精及胚胎发育过程进行观察研究。结果表明:(1)授粉后1h母本柱头上少量花粉开始萌发;授粉后4h大量花粉萌发,花粉管生长至柱头中部有胼胝质出现;授粉后6h花粉管生长至子房组织并有少量与胚珠结合;授粉后15h柱头中出现大量胼胝质,花粉管与胚珠结合数增多;授粉后24h胚珠周围出现多条花粉管,其中1条花粉管进入胚珠,部分进入胚囊的花粉管卷曲盘绕生长并产生胼胝质;精细胞与极核的融合主要发生在授粉后17~48h,与卵细胞融合主要于授粉后1~3d。(2)杂交石竹胚发育经过原胚、球形胚、棒状形胚、心形胚、鱼雷形胚和子叶形胚阶段。(3)杂交障碍表现为:只有游离的胚乳核而无胚发育的胚囊、合子未分裂、两极核未融合、球形胚败育。研究表明,杂交石竹存在受精前和受精后障碍,这是导致其结实率低的主要原因。     

The effect of spermidine and putrescine on the senescence of cut carnations

Blooms of Dianthus caryophyllus cv. Crowley Pink (carnations) were held under standard environmental conditions in a range of vase solutions. In the absence of preservative solution the senescence of the flower was characterized by a single sharp peak of ethylene production. Culture in preservative solution greatly extended the life of the bloom and also reduced the output of ethylene. In addition, there was great variability between individual blooms in the timing, the extent and the pattern of ethylene production. Instead of a single peak some blooms had two or even three small peaks which, in some cases, were separated by intervals of several days. Senescence of these flowers was also characteristic of blooms that were not producing ethylene in that the petals often did not inroll. Putrescine and spermidine, when added to the culture solutions, did not delay the onset of senescence, nor did they inhibit ethylene production. In fact, both additives resulted in the earlier production of ethylene and shorter longevity when applied in conjunction with preservatives. Their effects were similar, but less marked when they were applied alone. Although polyamines have been reported to delay senescence in a number of tissues, spermine and putrescine did not have a protective effect in carnation flowers; indeed, in some treatments they advanced senescence.     

Influence of 2,3,5-Triiodobenzoic Acid and 1-N-Naphthylphthalamic Acid on Indoleacetic Acid Transport in Carnation Cuttings: Relationship with Rooting

³H-IAA transport in excised sections of carnation cuttings was studied by using two receiver systems for recovery of transported radioactivity: agar blocks (A) and wells containing a buffer solution (B). When receivers were periodically renewed, transport continued for up to 8 h and ceased before 24 h. If receivers were not renewed, IAA transport decreased drastically due to immobilization in the base of the sections. TIBA was as effective as NPA in inhibiting the basipetal transport irrespective of the application site (the basal or the apical side of sections). The polarity of IAA transport was determined by measuring the polar ratio (basipetal/acropetal) and the inhibition caused by TIBA or NPA. The polar ratio varied with receiver, whereas the inhibition by TIBA or NPA was similar. Distribution of immobilized radioactivity along the sections after a transport period of 24 h showed that the application of TIBA to the apical side or NPA to the basal side of sections, increased the radioactivity in zones further from the application site, which agrees with a basipetal and acropetal movement of TIBA and NPA, respectively. The existence of a slow acropetal movement of the inhibitor was confirmed by using ³H-NPA. From the results obtained, a methodological approach is proposed to measure the variations in polar auxin transport. This method was used to investigate whether the variations in rooting observed during the cold storage of cuttings might be related to changes in polar auxin transport. As the storage period increased, a decrease in intensity and polarity of auxin transport occurred, which was accompanied by a delay in the formation and growth of adventitious roots, confirming the involvement of polar auxin transport in supplying the auxin for rooting. Received April 19, 1999; accepted December 2, 1999     

添加物及植物组合对瞿麦幼苗铅吸收、生长及品质的影响

通过盆栽试验研究了接种丛枝菌根真菌(AM真菌)、种植共生植物(三叶草)、添加重金属螯合物EDTA和磷肥磷酸钙对铅污染下瞿麦生长和品质的影响,为科学种植中药材提供理论依据.结果表明: 接种AM真菌可以显著抑制铅的吸收(P＜0.05),促进根部发育,使根冠比增大,且活性成分累积最多,大黄素含量为6.5 mg·g^-1;与三叶草共生后,抑制铅吸收的效果不佳且药材品质下降,大黄素含量低于对照组,降至3.2 mg·g^-1.但在AM真菌共同参与后瞿麦的生长量和活性成分有所增加,铅含量达到最低,低至1.3 mg·g^-1;添加重金属螯合剂会使瞿麦生长量下降并促进根部对铅的吸收,铅含量最高可达340.0 mg·g^-1;磷酸钙可固定土壤中其他重金属元素,故更适宜在复合型污染时使用.综合考虑,在保护中药材安全、品质方面AM真菌具有良好的应用价值.