Study on Anthocyanidin Extraction from Flower of Rhododendron L. and Its Stability

[Objective] The aim of this study was to further investigate and utilize the natural anthocyanidin from Rhododendron L. [Method] Using Rhododendron L. with four different colors including white, pink, red and purple as the test materials, its anthocyanidin extraction methods were analyzed in this study to evaluate the spectral properties and its stability of various anthocyanidin. [Result] The anthocyanidin from Rhododendron L. was perfectly extracted by methanol with 1% concentrated hydrochloric acid (V/V) and had better stability in this extraction solution. The further experiment in vitro indicated that the anthocyanidin from Rhododendron L. became stable with pH value of 0-3, but could not resistant to high temperature or strong light, and the alkaline condition had also great effects on its stability. [Conclusion] The methanol with 1% concentrated hydrochloric acid (V/V) has the best effect for extraction, and the anthocyanidin from Rhododendron L. is more stable in low temperature, weak light and acid conditions.

杜鹃花属(Rhododendron L.)植物DNA条形码鉴定——参考序列库、分子标记与栽培品种野生亲本溯源

杜鹃花属(Rhododendron L.)植物作为重要观赏和中药植物,市场前景广阔,发展潜力巨大。本文通过归纳杜鹃花属植物的分类、鉴定相关问题,发现利用DNA条形码可有效解决杜鹃花属植物(品种)的鉴定问题。基于杜鹃花属植物的具体情况,本文对序列参考数据库建设、分子标记开发以及杜鹃花品种的野生亲本溯源三方面内容进行综述,并结合GenBank中1311条杜鹃花属DNA序列及作者测序获得的463条序列,初步建立杜鹃花属DNA条形码数据库,并通过非加权组平均法利用matK与psbA-trnH序列联合建立UPGMA树,部分杜鹃花属栽培品种可与野生种聚为一支。综合说明DNA条形码技术可用于对杜鹃花属栽培品种开展初步的野生亲本溯源。

L形冷弯薄壁型钢组合钢管混凝土柱轴压性能

为解决异形钢管混凝土柱阴角对其承载力的影响,提出了用1个冷弯薄壁方钢管和2个U形钢管焊接成L形钢管,内填充混凝土形成L形冷弯薄壁型钢组合钢管混凝土柱形式。设计了5组共10根试件的轴压试验,并开展了有限元参数分析,研究了钢管厚度、U形管外伸长度和钢材强度等参数对构件承载力和延性等力学性能的影响。结果表明:该类试件的主要破坏形态为中上部局部鼓曲破坏,适量增大U形管外伸长度可以提高承载力,但增大到一定程度之后易发生弯扭破坏;承载力和延性随着钢管厚度和钢材强度增大而增加;混凝土强度对构件的初始刚度和峰值荷载的影响都很小,但对曲线下降段的影响较大;试件端部和中部截面阴角处的混凝土应力值比各边中部更大,说明采用U形钢管与方钢管组合的方式改善了阴角处钢管对混凝土约束普遍较弱的问题。基于“统一理论”,给出了两种承载力建议计算公式,与试验结果吻合较好,两种计算方法在0.44~1.94的约束效应系数范围内具有良好的适用性。

Island biogeography theory and the habitat heterogeneity jointly explain global patterns of Rhododendron diversity

Mountain biodiversity is of great importance to biogeography and ecology.However,it is unclear what ecological and evolutionary processes best explain the generation and maintenance of its high levels of species diversity.In this study,we determined which of six common hypotheses(e.g.,climate hypotheses,habitat heterogeneity hypothesis and island biogeography theory)best explain global patterns of species diversity in Rhododendron.We found that Rhododendron diversity patterns were most strongly explained by proxies of island biogeography theory(i.e.,mountain area)and habitat heterogeneity(i.e.,elevation range).When we examined other relationships important to island biogeography theory,we found that the planimetric area and the volume of mountains were positively correlated with the Rhododendron diversity,whereas the‘mountains-to-mainland’distance was negatively correlated with Rhododendron diversity and shared species.Our findings demonstrate that Rhododendron diversity can be explained by island biogeography theory and habitat heterogeneity,and mountains can be regarded as islands which supported island biogeography theory.

RAD-seq data reveals robust phylogeny and morphological evolutionary history of Rhododendron

Rhododendron is famous for its high ornamental value.However,the genus is taxonomically difficult and the relationships within Rhododendron remain unresolved.In addition,the origin of key morphological characters with high horticulture value need to be explored.Both problems largely hinder utilization of germplasm resources.Most studies attempted to disentangle the phylogeny of Rhododendron,but only used a few genomic markers and lacked large-scale sampling,resulting in low clade support and contradictory phylogenetic signals.Here,we used restriction-site associated DNA sequencing(RAD-seq)data and morphological traits for 144 species of Rhododendron,representing all subgenera and most sections and subsections of this species-rich genus,to decipher its intricate evolutionary history and reconstruct ancestral state.Our results revealed high resolutions at subgenera and section levels of Rhododendron based on RAD-seq data.Both optimal phylogenetic tree and split tree recovered five lineages among Rhododendron.Subg.Therorhodion(cladeⅠ)formed the basal lineage.Subg.Tsutsusi and Azaleastrum formed cladeⅡand had sister relationships.CladeⅢincluded all scaly rhododendron species.Subg.Pentanthera(cladeⅣ)formed a sister group to Subg.Hymenanthes(cladeⅤ).The results of ancestral state reconstruction showed that Rhododendron ancestor was a deciduous woody plant with terminal inflorescence,ten stamens,leaf blade without scales and broadly funnelform corolla with pink or purple color.This study shows significant distinguishability to resolve the evolutionary history of Rhododendron based on high clade support of phylogenetic tree constructed by RAD-seq data.It also provides an example to resolve discordant signals in phylogenetic trees and demonstrates the application feasibility of RAD-seq with large amounts of missing data in deciphering intricate evolutionary relationships.Additionally,the reconstructed ancestral state of six important characters provides insights into the innovation of key characters in Rhododendron.

Network Pharmacological Analysis of Action Mechanism of Mongolian Medicine Rhododendron micranthum Turcz. in Treating Lung Cancer

[Objectives]To explore the action mechanism of Mongolian medicine Rhododendron micranthum Turcz.on lung cancer by network pharmacology.[Methods]Based on the high-throughput experiment and reference database(HERB)of traditional Chinese medicine,component target database(Swiss ADME),small molecule drug target prediction online platform(SWISS Target Prediction),human gene business card database(GENECARD),the database of genes and mutation sites related to human diseases(DISGENET)and other databases,the target genes of drugs and diseases were screened out.Venny software was used for obtaining the target intersection of active components of the Mongolian medicine R.micranthum Turcz.and the lung cancer,a CytoNCA plug-in in cytoscape 3.10.0 software was used for screening candidate core target genes,and related effective components were obtained in a reverse direction.A drug-active ingredient-gene-disease regulation network was established,a protein-protein interaction(PPI)network was established by means of the STRING database to screen core genes,and common targets were screened by the David database.Gene Ontology(GO)function and Kyoto Encyclopedia of Genes and Genomes(KEGG)were used for enrichment analysis.[Results]There were 13 effective components of Mongolian medicine R.micranthum Turcz.for treating lung cancer and 115 drug disease intersection target genes.Core genes affecting the disease included SRC,HSP90AB1,EGFR,AKT1,and ERBB2.GO functional enrichment analysis involved 462 items of biological processes,64 items of cellular components and 126 items of molecular functions.Enrichment analysis of KEGG signaling pathways screened out cancer pathways,endocrine resistance,PI3K-Akt signaling pathways,proteoglycans in cancer and other signaling pathways.[Conclusions]Mongolian medicine R.micranthum Turcz.can inhibit the proliferation of lung cancer cells from multiple targets and pathways,and the results of network pharmaceutical analysis provide a theoretical basis for further experimental research.

甘蓝型油菜BnaSLY1基因进化分析及功能研究

赤霉素调控植物表皮细胞增长、茎叶伸长以及株型建成。拟南芥SLY1属于F-box蛋白,它通过靶向泛素化赤霉素信号转导路径的负向调控因子——DELLA蛋白来影响植物生长发育,然而油菜中BnaSLY1的基因功能尚未揭示。本研究对BnaSLY1进行了表达特征和进化树分析,利用CRISPR/Cas9技术创制了BnaSLY1不同拷贝数的突变体,结合RNA-Seq技术对BnaSLY1的生物学功能及其对油菜生长发育的影响进行了研究。结果表明,甘蓝型油菜Westar中有2个SLY1同源拷贝BnaA01.SLY1和BnaA06.SLY1,它们表达模式基本相同,为组成型表达基因,其蛋白定位在细胞核,且在不同的油菜品种及十字花科植物间序列保守。与对照相比,单突bnaa01sly1和bnaa06sly1开花时间推迟,株高显著降低,而双突bnasly1还表现出深绿色光叶表型,叶片厚度增加,开花期比单突进一步推迟,株高也进一步降低。RNA-Seq结果显示,双突与Westar之间的差异表达基因显著富集在生长素信号转导路径以及蜡质合成通路,多个开花时间相关基因表达也发生显著变化。本研究表明, BnaSLY1除影响植物株高和开花时间等生长发育进程,还影响表皮蜡质合成,为探索赤霉素信号转导路径在甘蓝型油菜生长发育中的重要作用奠定了理论基础。

缺血性脑卒中铁死亡特征基因NFE2L2的鉴定与验证

背景:铁死亡与缺血性脑卒中的发病密切相关,靶向铁死亡是一种治疗缺血性脑卒中有前景的方案,但具体调控靶点尚不明确。目的:通过生物信息学和机器学习方法筛选缺血性脑卒中铁死亡相关特征基因,并通过细胞实验进行验证,探讨铁死亡在缺血性脑卒中的作用。方法:基于GEO数据库和FerrDb数据库选取符合条件的缺血性脑卒中相关数据集和铁死亡表达数据集,通过t检验筛选铁死亡相关差异基因。对铁死亡相关差异基因进行GO功能富集分析与KEGG信号通路富集分析。通过PPI网络分析和机器学习筛选缺血性脑卒中铁死亡的特征基因,利用ROC分析和GSEA分析探究特征基因的准确性和生物功能。然后进行细胞实验,将HT22细胞分为对照组与缺血性脑卒中组,对照组不作任何干预,缺血性脑卒中组加入0.1 mol/L的H_(2)O_(2)干预24 h诱导细胞氧化应激和铁死亡,通过实时荧光定量RT-PCR和Western Blot验证铁死亡的发生和特征基因表达。结果与结论:(1)共获取45个铁死亡相关差异基因,GO和KEGG富集分析发现差异基因与氧化应激、自噬、铁死亡、脂肪细胞因子信号通路和线粒体代谢密切相关。(2)通过PPI网络中的MCODE插件和cytoHubba插件与机器学习中的LASSO算法和SVM-RFE算法共鉴定出1个铁死亡特征基因核因子E2相关因子2(nuclear factor erythroid 2-related factor 2,NFE2L2)。(3)对NFE2L2进行ROC曲线分析,发现在训练集和验证集中构建的诊断预测模型具有良好的准确性与特异性;对NFE2L2进行GSEA分析,发现特征基因通过免疫、炎症反应、氨基酸代谢及神经因子调控等方面参与缺血性脑卒中发病机制的调控。(4)细胞实验的RT-PCR和Western Blot分析表明,与对照组对比,缺血性脑卒中组中的酰基辅酶A合成酶长链家族成员4 mRNA和蛋白表达水平显著增高(P<0.05),谷胱甘肽过氧化物酶4 mRNA和蛋白表达水平显著降低(P<0.05);与对照组对比,缺血性脑卒中组中特征基因NFE2L2 mRNA和蛋白表达水平显著增高(P<0.05)。(5)上述结果证实,缺血性脑卒中与铁死亡密切相关,靶向特征基因NFE2L2可以为研究和治疗缺血性脑卒中提供一定的思路与方向。

基于整车关键尺寸L114的平台架构研究

为了研究最优的整车平台架构策略,本文详细阐述平台架构的概念,定义了架构关键尺寸L114,列举国内某车企基于关键尺寸L114一致的平台拓展尺寸链,并梳理该平台架构乘员舱架构件具体的策略,通过对比分别采取平台车型关键尺寸L113一致与平台车型关键尺寸L114一致时的平台通用化率,发现2种策略的平台通用化率基本一致。同时结合整车安全碰撞策略进行分析,结果表明,若采用关键尺寸L114更具优势,该策略下同平台车型仅需满足高车型的碰撞策略,即可同步满足其他车型的侧碰要求,有效降低平台开发成本。

Effects of P301L-TAU on post-translational modifications of microtubules in human iPSC-derived cortical neurons and TAU transgenic mice

TAU is a microtubule-associated protein that promotes microtubule assembly and stability in the axon.TAU is missorted and aggregated in an array of diseases known as tauopathies.Microtubules are essential for neuronal function and regulated via a complex set of post-translational modifications,changes of which affect microtubule stability and dynamics,microtubule interaction with other proteins and cellular structures,and mediate recruitment of microtubule-severing enzymes.As impairment of microtubule dynamics causes neuronal dysfunction,we hypothesize cognitive impairment in human disease to be impacted by impairment of microtubule dynamics.We therefore aimed to study the effects of a disease-causing mutation of TAU(P301L)on the levels and localization of microtubule post-translational modifications indicative of microtubule stability and dynamics,to assess whether P301L-TAU causes stability-changing modifications to microtubules.To investigate TAU localization,phosphorylation,and effects on tubulin post-translational modifications,we expressed wild-type or P301L-TAU in human MAPT-KO induced pluripotent stem cell-derived neurons(i Neurons)and studied TAU in neurons in the hippocampus of mice transgenic for human P301L-TAU(p R5 mice).Human neurons expressing the longest TAU isoform(2N4R)with the P301L mutation showed increased TAU phosphorylation at the AT8,but not the p-Ser-262 epitope,and increased polyglutamylation and acetylation of microtubules compared with endogenous TAU-expressing neurons.P301L-TAU showed pronounced somatodendritic presence,but also successful axonal enrichment and a similar axodendritic distribution comparable to exogenously expressed 2N4R-wildtype-TAU.P301L-TAU-expressing hippocampal neurons in transgenic mice showed prominent missorting and tauopathy-typical AT8-phosphorylation of TAU and increased polyglutamylation,but reduced acetylation,of microtubules compared with non-transgenic littermates.In sum,P301L-TAU results in changes in microtubule PTMs,suggestive of impairment of microtubule stability.This is accompanied by missorting and aggregation of TAU in mice but not in i Neurons.Microtubule PTMs/impairment may be of key importance in tauopathies.

Loss-of-function mutations of microRNA-142-3p promote ASH1L expression to induce immune evasion and hepatocellular carcinoma progression

BACKGROUND Hepatocellular carcinoma(HCC)has been a pervasive malignancy throughout the world with elevated mortality.Efficient therapeutic targets are beneficial to treat and predict the disease.Currently,the exact molecular mechanisms leading to the progression of HCC are still unclear.Research has shown that the microRNA-142-3p level decreases in HCC,whereas bioinformatics analysis of the cancer genome atlas database shows the ASH1L expression increased among liver tumor tissues.In this paper,we will explore the effects and mechanisms of microRNA-142-3p and ASH1L affect the prognosis of HCC patients and HCC cell bioactivity,and the association between them.AIM To investigate the effects and mechanisms of microRNA-142-3p and ASH1L on the HCC cell bioactivity and prognosis of HCC patients.METHODS In this study,we grouped HCC patients according to their immunohistochemistry results of ASH1L with pathological tissues,and retrospectively analyzed the prognosis of HCC patients.Furthermore,explored the roles and mechanisms of microRNA-142-3p and ASH1L by cellular and animal experiments,which involved the following experimental methods:Immunohistochemical staining,western blot,quantitative real-time-polymerase chain reaction,flow cytometric analysis,tumor xenografts in nude mice,etc.The statistical methods involved in this study contained t-test,one-way analysis of variance,theχ^(2)test,the Kaplan-Meier approach and the log-rank test.RESULTS In this study,we found that HCC patients with high expression of ASH1L possess a more recurrence rate as well as a decreased overall survival rate.ASH1L promotes the tumorigenicity of HCC and microRNA-142-3p exhibits reduced expression in HCC tissues and interacts with ASH1L through targeting the ASH1L 3′untranslated region.Furthermore,microRNA-142-3p promotes apoptosis and inhibits proliferation,invasion,and migration of HCC cell lines in vitro via ASH1L.For the exploration mechanism,we found ASH1L may promote an immunosuppressive microenvironment in HCC and ASH1L affects the expression of the cell junction protein zonula occludens-1,which is potentially relevant to the immune system.CONCLUSION Loss function of microRNA-142-3p induces cancer progression and immune evasion through upregulation of ASH1L in HCC.Both microRNA-142-3p and ASH1L can feature as new biomarker for HCC in the future.

载富血小板血浆水凝胶对L929细胞氧化损伤的保护作用

背景:在慢性创面愈合过程中,活性氧的过量生成可能会损害L929成纤维细胞的功能,从而延缓创面修复,因此保护成纤维细胞免受氧化应激的影响对于促进创面愈合非常重要。目的:评估羧甲基壳聚糖-氧化硫酸软骨素/富血小板血浆(CMC-OCS/PRP)水凝胶对H2O2刺激下L929细胞的保护作用。方法:制备CMC-OCS/PRP水凝胶,表征水凝胶的微观形貌、降解性能、清除H2O2与羟基自由基的能力及生物相容性。取生长状态良好的L929细胞,分5组培养:对照组常规培养,H2O2组加入H2O2,CMC-OCS组加入羧甲基壳聚糖-氧化硫酸软骨素水凝胶浸提液+H2O2,PRP组加入富血小板血浆凝胶浸提液+H2O2,CMC-OCS/PRP组加入羧甲基壳聚糖-氧化硫酸软骨素/富血小板血浆水凝胶浸提液处理+H2O2,每组先加入水凝胶浸提液处理6 h,然后再加入H2O2处理24 h。培养结束后,检测细胞活性氧及丙二醛水平,细胞凋亡、胶原纤维Ⅰ蛋白表达。在加入H2O2的情况下,将上述水凝胶浸提液分别与L929成纤维细胞直接或间接共培养36 h,通过划痕实验、Transwell小室实验检测细胞的迁移能力。结果与结论:①CMC-OCS/PRP水凝胶具有均匀相互关联的多孔结构及良好的降解能力,体外可有效清除H2O2与羟基自由基,并且具有良好的生物相容性;②与对照组比较,H2O2组细胞凋亡率、细胞活性氧与丙二醛水平升高(P<0.05),细胞铺展面积减少(P<0.05),胶原纤维Ⅰ蛋白表达无明显变化(P>0.05);与H2O2组比较,CMC-OCS组、CMC-OCS/PRP组细胞活性氧水平降低(P<0.05),PRP组、CMC-OCS组、CMCOCS/PRP组丙二醛水平降低(P<0.05)、细胞铺展面积增加(P<0.05),CMC-OCS/PRP组细胞凋亡率降低(P<0.05),PRP组、CMC-OCS组、CMCOCS/PRP组胶原纤维Ⅰ蛋白表达增加(P<0.05);③与对照组比较,H2O2组细胞迁移数量减少(P<0.05),迁移面积无明显变化(P>0.05);与H2O2组比较,PRP组、CMC-OCS组、CMC-OCS/PRP组细胞迁移数量、迁移面积均增加(P<0.05),并且以CMC-OCS/PRP组增加最显著;④在氧化应激条件下,CMC-OCS/PRP水凝胶可改善成纤维细胞的迁移能力,抗细胞凋亡并保护细胞伸展功能。

Recombinant chitinase-3-like protein 1 alleviates learning and memory impairments via M2 microglia polarization in postoperative cognitive dysfunction mice

Postoperative cognitive dysfunction is a seve re complication of the central nervous system that occurs after anesthesia and surgery,and has received attention for its high incidence and effect on the quality of life of patients.To date,there are no viable treatment options for postoperative cognitive dysfunction.The identification of postoperative cognitive dysfunction hub genes could provide new research directions and therapeutic targets for future research.To identify the signaling mechanisms contributing to postoperative cognitive dysfunction,we first conducted Gene Ontology and Kyoto Encyclopedia of Genes and Genomes pathway enrichment analyses of the Gene Expression Omnibus GSE95426 dataset,which consists of mRNAs and long non-coding RNAs differentially expressed in mouse hippocampus3 days after tibial fracture.The dataset was enriched in genes associated with the biological process"regulation of immune cells,"of which Chill was identified as a hub gene.Therefore,we investigated the contribution of chitinase-3-like protein 1 protein expression changes to postoperative cognitive dysfunction in the mouse model of tibial fractu re surgery.Mice were intraperitoneally injected with vehicle or recombinant chitinase-3-like protein 124 hours post-surgery,and the injection groups were compared with untreated control mice for learning and memory capacities using the Y-maze and fear conditioning tests.In addition,protein expression levels of proinflammatory factors(interleukin-1βand inducible nitric oxide synthase),M2-type macrophage markers(CD206 and arginase-1),and cognition-related proteins(brain-derived neurotropic factor and phosphorylated NMDA receptor subunit NR2B)were measured in hippocampus by western blotting.Treatment with recombinant chitinase-3-like protein 1 prevented surgery-induced cognitive impairment,downregulated interleukin-1βand nducible nitric oxide synthase expression,and upregulated CD206,arginase-1,pNR2B,and brain-derived neurotropic factor expression compared with vehicle treatment.Intraperitoneal administration of the specific ERK inhibitor PD98059 diminished the effects of recombinant chitinase-3-like protein 1.Collectively,our findings suggest that recombinant chitinase-3-like protein 1 ameliorates surgery-induced cognitive decline by attenuating neuroinflammation via M2 microglial polarization in the hippocampus.Therefore,recombinant chitinase-3-like protein1 may have therapeutic potential fo r postoperative cognitive dysfunction.

Autism spectrum disorder:difficulties in diagnosis and microRNA biomarkers

We performed a PubMed search for microRNAs in autism spectrum disorder that could serve as diagnostic biomarkers in patients and selected 17 articles published from January 2008 to December 2023,of which 4 studies were performed with whole blood,4 with blood plasma,5 with blood serum,1 with serum neural cell adhesion molecule L1-captured extracellular vesicles,1 with blood cells,and 2 with peripheral blood mononuclear cells.Most of the studies involved children and the study cohorts were largely males.Many of the studies had performed microRNA sequencing or quantitative polymerase chain reaction assays to measure microRNA expression.Only five studies had used real-time polymerase chain reaction assay to validate microRNA expression in autism spectrum disorder subjects compared to controls.The microRNAs that were validated in these studies may be considered as potential candidate biomarkers for autism spectrum disorder and include miR-500a-5p,-197-5p,-424-5p,-664a-3p,-365a-3p,-619-5p,-664a-3p,-3135a,-328-3p,and-500a-5p in blood plasma and miR-151a-3p,-181b-5p,-320a,-328,-433,-489,-572,-663a,-101-3p,-106b-5p,-19b-3p,-195-5p,and-130a-3p in blood serum of children,and miR-15b-5p and-6126 in whole blood of adults.Several important limitations were identified in the studies reviewed,and need to be taken into account in future studies.Further studies are warranted with children and adults having different levels of autism spectrum disorder severity and consideration should be given to using animal models of autism spectrum disorder to investigate the effects of suppressing or overexpressing specific microRNAs as a novel therapy.

Investigation on Natural Resources of Rhododendron at Muwang

Based on the field investigations, the natural status of Muwang was introduced, then the morphological characteristics, community traits, growth and development habits and biological characteristics, distributive pattern, developing and utilizing status of Rhododendron germplasm were analyzed. The suggestion was advanced about strengthening the work of on-site conservation, so as to insure the resources can be utilized forever.

Infrageneric and Sectional Relationships in the Genus Rhododendron (Ericaceae) Inferred from ITS Sequence Data

The internal transcribed spacer (ITS) regions of nuclear ribosomal DNA (including 5.8S rRNA) of 15 Rhododendron, species, representing most sections of the genus, one Ledum species and Cassiope fastigiata were sequenced. Together with the ITS sequences of 13 selected Rhododendron species and Bejaria racemosa downloaded from GenBank, we explored the infrageneric and sectional relationships of this important North Temperate genus by employing maximum-parsimony analysis using PAUP software. C. fastigiata and B. racemosa were designated as outgroups. The ITS-based tree inferred that: (1) Rhododendron was a well-supported monophyletic group, while subg. Therorhodion was basal to the rest of the genus; (2) Ledum was a member of Rhododendron, and its close relationship with the lepidote rhododendron was confirmed; (3) the lepidote rhododendron plus Ledum formed a strongly-supported monophyletic clade which was sister to the rest of the elepidote rhododendron; (4) the elepidote rhododendron formed a weakly-supported clade within which the monophyly of subg. Hynwrianthes and subg. Tsutsusi were strongly supported, while subg. Pentanthera and subg. Azaleastrum were polyphyletic; and (5) the monophyly of sect. Choniastnini, (subg. Azaleastrum) was strongly-supported, while subg. Tsutsusi could be sister to a weakly-supported clade composed of two sampled species of sect. Azaleastrum (subg. Azaleastrum) together with R. sentibarbatum, of subg. Mumeazalea.

Uniform Design for Optimizating Regeneration Shoots Directly from Tender Leaves and Plant Regeneration System of Rhododendron chrysanthum Pall.

[Objective] The experimental was aimed to screen the optimum regeneration shoot induction media and rooting media for tender leaves of Rhododendron chrysanthum Pall.[Method] The tender leaves of Rhododendron chrysanthum Pall were taken as explants to select the optimum bud induction media and rooting media through uniform design and the screening results were verified.[Result] The optimum media for regeneration shoot of Rhododendron chrysanthum Pall contained 1/4 MS,3.70 mg/L ZT, 0.02 mg/L IAA and 1.00 mg/L KT and its induction rate was 95.5% and the rooting media contained modified MS, 0.10 mg/L IAA and 0.07 mg/L NAA and its rooting rate was 98%. [Conclusion] Through this experiment, regeneration systems for regeneration shoot and regenerated plant from tender leaves of Rhododendron chrysanthum Pall were created successfully.

EST-SSR Marker-based Genetic Diversity Analysis of Rhododendron simsii Germplasm in Guifeng Mountain

Rhododendron simsii（Ericaeae：Rhododendron） has high ornamental value and ecological value.In this study,7 pairs of novel EST-SSR markers were developed from the genomic sequence of R.simsii,and they were used to investigate the genetic diversity of 32 natural R.simsii samples from Guifeng Mountain,Hubei Province.Results showed that a total of 31 polymorphic bands were amplified with allele number per locus of 4.43.Mean values of heterozygosity（Ho） and expected heterozygosity（He） were 0.679 58 and 0.723 14,respectively.This research will not only enrich the existing SSR database,but also lay a foundation for subsequent studies about molecular marker-assisted breeding,genetic diversity analysis,genetic structure analysis and phylogenetic analysis.

羊踯躅(Rhododendron molle)生物学特性观察Ⅰ

本文采用直接观测法,对羊踯躅四年生种苗的根、茎、叶进行观察、测量、称量、计算,结果表明,羊踯躅的地下部分根颈部粗大,须根系庞大;茎分枝较少,萌生枝条和多年生枝同时并存,其生长量与落叶灌木玫瑰的生长量基本相似;叶着生在当年生枝条的顶端,叶片覆盖面积为80.2%,滞水量95.5%。

羊踯躅(Rhododendron molle)种子萌发试验

用不同的方法对羊踯躅种子进行萌发试验,结果表明羊踯躅风干的蒴果和脱粒在4℃低温贮存,其发芽率较高,用培养皿法可提高种子的发芽率和出苗率。