Phylogenetic analysis of the internal transcribed spacer region of Japanese Lilium species

 The DNA from 16 Lilium species and one variety endemic to or naturalized in Japan were obtained and their internal transcribed spacer regions of nuclear ribosomal DNA (nrDNA) were amplified by PCR and sequenced by cycle sequencing. Phylogenetic analysis of the ITS sequences supported the validity of Comber’s classification system. It has also provided molecular evidence for the transfer of Lilium dauricum to sect. Sinomartagon. The phylogenetic relationships revealed by ITS DNA analysis were supported by previously published crossability data. The molecular phylogeny of Japanese Lilium species was discussed with reference to the putative migration routes of these species. Received: 30 June 1998 / Accepted: 19 October 1998     

Immunoaffinity Chromatographic and ELISA determination of Cytokinins in Germinating Pollens in Lilium davidii Dach.

Liliurn davidii Duch. pollen was germinated in PEG-400. A rapid immunoaffinity chromatograpy system with high efficiency and specificity was adopted to isolate and purify trans-zeatin riboside (t-ZR) and isopentenyladenosine (ipA). Cytokinin determination was carried out by a highly sensitive enzyme linked immunosolvent assay (ELISA) for t-ZR and iPA respectively, t-ZR and iPA were present at the level of 10-8 g/g fr. wt in Lilium davidii pollen. After hydration, t-ZR content of pollen decreased slightly, while iPA increased remarkably, and the total amount of t-ZR and iPA underwent almost no change. Pollen tubes grew fast in the first 3 h after germination, and the amount of t-ZR and iPA also increased greatly both in pollen tubes and in germination medium. The increasing trend of t-ZR and iPA and growth rate of pollen tubes was basically synchronous.     

Transmission and Scanning Electron Microscopic Observations on the Plasmodesmatal Channels Between the Pollen Mother Cells of Lily

The regularity of the presence of plasmodesmata channels in the pollen mother cells of lily was studied by transmission and scanning electron microscopy. A few plasmodesmata channels can be recognized between the pollen mother cells at leptotene stage, which increase in number at zygotene and expand in width at synizesis and they lie in the range 0.5—1 μm. Massive chromatin substance are transferred from one pollen mother cell to another during synizesis. The pre-existing plasmodesmate channels close again at late pachytene. There are no channels from metaphase Ⅰ to tetrad stage. Finally, the relation between the presence of plasmodesmata channels, synizesis and cytomixis were discussed.     

A Preliminary Observation on B-Chromosomes of Regal lily

The authors have observed t he karyotypes of the Regal Lily with B-chromosome and without B-chromosomes as well as their Giemsa C-bands. The results show that the karyotypes with B-chromosomes are not obviously different from those without B- chromosomes. B-chromosome is additional and all of them are heterochromatization. But segregation of B-chromosome at anaphase Ⅰ and Ⅱ in meiosis is irregular, and about half of the pollen grains contain B-chromosomes. In the progenies by open pollination the segre, gation of 0B and lB appear in the ratio near 1:1 and the offsprings with 2B-chromosomes only average 2.6 percent.     

Cytochemical Localization of Adenosine Triphosphataes Activity During Cytomixis in Pollen Mother Cells of David Lily and Its Relation to the Intercellular Migrating Chromatin Substance

Standard lead precipitation procedures have been used to examine the localization of ATPase activity during cytomixis in pollen mother cells of Lilium davidii var. willmottiae (Wilson) Roffill. Before cytomixis, cells at this stage of development show ATPase activity on plasma membrane, in the endoplasmic reticulum, dictyosomes, plastids, plasmodesmata, and in part of the groundplasm; however, there is no ATPase activity on the chromatin and nucleolus. During cytomixis, the chromatin substance begin to transfer from one cell to an adjacent cell, reaction product indicating ATPase activity is observed associated with the chromatin and nucleolus. ATPase activity is also found with the cistenae of both endoplasmic reticulum and dictyosomes, and some plastids. There is no deposition of ATPase reaction product associated with the plasm membrane and intercellular spaces. After cytomixis, the chromatin is little or no deposition of enzyme reaction product. ATPase activity, however, is consistenlly found within the intercellular space and on the plasm membrane, and also occur in the endoplasmic reticulum, dictyosome and plastid. The presence or absence of ATPase activity in the cell structure of pollen mother cells before, during or after eytomixis is discussed in relation to the active uptake or export of water for short-distance transport. It is also suggested that the intensive ATPase activity in the nucleus during cytomixis of pollen mother cells is evidence for a transport system involved in the active movement of the intercellular migrating ebromatin substance.     

Disturbance of endomembrane trafficking by brefeldin A and calyculin A reorganizes the actin cytoskeleton of <Emphasis Type="Italic">Lilium longiflorum</Emphasis> pollen tubes

We investigated the effect of brefeldin A on membrane trafficking and the actin cytoskeleton of pollen tubes of Lilium longiflorum with fluorescent dyes, inhibitor experiments, and confocal laser scanning microscopy. The formation of a subapical brefeldin A-induced membrane aggregation (BIA) was associated with the formation of an actin basket from which filaments extended towards the tip. The orientation of these actin filaments correlated with the trajectories of membrane material stained by FM dyes, suggesting that the BIA-associated actin filaments are used as tracks for retrograde transport. Analysis of time series indicated that these tracks (actin filaments) were either stationary or glided along the plasma membrane towards the BIA together with the attached membranes or organelles. Disturbance of the actin cytoskeleton by cytochalasin D or latrunculin B caused immediate arrest of membrane trafficking, dissipation of the BIA and the BIA-associated actin basket, and reorganization into randomly oriented actin rods. Our observations suggest that brefeldin A causes ectopic activation of actin-nucleating proteins at the BIA, resulting in retrograde movement of membranes not only along but also together with actin filaments. We show further that subapical membrane aggregations and actin baskets supporting retrograde membrane flow can also be induced by calyculin A, indicating that dephosphorylation by type 2 protein phosphatases is required for proper formation of membrane coats and polar membrane trafficking.     

凤凰百合——百合属一新种

凤凰百合　新种　图 1 ,图版 Lilium floridum　 J.L.Ma et Y.J.Li,sp.nov.Fig.1 ,Plate Species nova L.leichtlinii var.maximowiczii(Regel) Baker affinis,sed caulibus,rachidibus et pedicellis longitudinaliter porcatis et albo-tomentosis;foliis,bracteis etalabastris albo-tomentosis;tepalis supra disperso maculatis;maculis atropurpureis con-vexis bene differt.Secunde,species nova differt a L.amabile Palibin,cuius caulis rachis et pedicellusnon albo-tomentosus,macula super tepalum non convexa;pro…     

Effect of Extracellular Calmodulin on the Cytosolic Ca2+ Concentration in Lily Pollen Grains

Using Lilium davidii Duchartre pollen as material, the calcium ion-fluorescence indicator fluo-3AM was loaded successfully into the pollen grains by low temperature loading method. Laser confocal scanning microscopy was used to study the effect of extracellular calmodulin on intracellular calcium. It is found that the purified exogenous calmodulin could elevate the intracellular calcium ion concentration, and the effect was correlated with the concentration of exogenous calmodulin to a certain extent. Cell membrane nonpermeable inhibitor of calmodulin, W 7-agarose, and the anti-serum of calmodulin could decrease the cytosolic calcium level. The results show that the endogenous extracellular calmodulin may play an important role in maintaining and increasing the cytosolic calcium level in pollen grain cell.     

K+ Channels in the Plasma Membrane of Lily Pollen Protoplasts

Using the patch-clamp technique K⁺ channels could be observed in the plasma membrane of protoplasts from pollen grains of Lilium longiflorum. With depolarizing membrane potentials the open probability of the different K⁺ channels increased. Two K⁺ channel populations occurring occasionally had a single channel conductance of 120 pS and 42 pS, respectively. The most often observed K⁺ channel had a single channel conductance of 19 pS which showed an increase of channel activity with increasing free cytoplasmic Ca²⁺ concentration. This channel population might be involved in the pathway of endogenous transcellular K⁺ currents which are activated during pollen tube tip extension.     

一种崂山百合的离体快速繁殖技术

以崂山百合的鳞茎为外植体,通过植物组织培养技术成功获得了再生植株,建立了该植物的快速无性繁殖体系。整个组织培养过程分为芽的诱导、根的诱导和移栽三个阶段。适合芽诱导的最佳外植体是:鳞茎基部,最适培养基是:1/2 MS+0.5 mg/LNAA+0.8 mg/L KT+3%蔗糖。用于生根的最佳培养基是:1/2 MS+0.4 mg/L IBA+3%蔗糖。最适培养温度为25℃.不同基质对组培苗移栽的成活率有一定影响,其中在珍珠岩上的移栽成活率为98%。