野生花卉胭脂花(Primula maximowiczii)组培体系的建立

以胭脂花(Primula maximowiczii)的种子为外植体,利用单因子和正交试验设计,通过对种子表面消毒、丛生芽诱导和增殖、生根、炼苗和移栽等技术环节的研究,筛选出各培养阶段的最佳培养基配方和培养条件,建立了胭脂花无菌培养体系。结果表明:种子表面消毒的最佳方法是以6%NaClO消毒6 min;最佳丛生芽诱导增殖培养基为MS+6-BA1.0 mg.L-1H+NAA0.20 mg.L-1,丛生芽增殖系数为3.82;生根培养中,IBA比NAA和IAA更有利于根的形成和生长,IBA的最佳质量浓度为0.02～0.05 mg.L-1,生根率达94%以上。

安徽报春花属Primula L.植物的发现和研究史

报春花属 Primu1a L.植物全世界共产500余种,我国种类最为丰富,有293种,21亚种,18变种,约占世界总数的60%。主要分布集中在喜马拉雅山至云南、四川西部一带,这一地区是本属植物的现代分布中心。本属植物大多花期较早,通常早春就可开花,中名报春花和学名 Primula 就具有这种含义。属植物不仅种类繁多,许多种类花色艳丽,是重要的园庭观赏植物。本世纪初,欧、美许多国家多次派人来我国采集,将种苗运回本国栽培。

Primula pengzhouensis (Primulaceae), a new species from Sichuan, southwestern China

A new species, Primula pengzhouensis(Primulaceae), from central Sichuan, China, is described and illustrated. It is assigned to Primula sect. Aleuritia subsect. Yunnanensis, and is most similar to P. socialis,but can be easily distinguished from that species by its much larger flower and elliptic or ovate-elliptic leaves.

胭脂花(Primula maximowiczii)种子消毒及无菌播种的研究

为获得胭脂花无菌苗,本试验利用单因子试验和正交试验设计,探索了不同消毒方法、基本培养基、大量元素浓度和蔗糖对无菌种子萌发和幼苗生长的影响。结果表明:种子表面消毒的最佳方法是以6%NaClO消毒6min;最佳无菌萌发基本培养基是MS或B5培养基;MS培养基大量元素浓度在1倍时,最适宜于胭脂花种子萌发;蔗糖含量在15～45 g/L范围内对发芽率的影响差异不明显。

鄂报春(Primula obconica Hance)亲缘地理学的初步研究

为了获得鄂报春(PrimulaobconicaHance)的种内亲缘地理学信息,对叶绿体基因组上非编码区trnL-trnF的序列变异进行了分析。获得的20个单倍型具有一定的地理分布结构。对这些单倍型进行系统发育分析,得到了三个主要分支:东部分支广泛分布于中国东南部、湖南、湖北、四川和云南;四川分支分布于四川西部;云南分支分布于云南西北部。东部分支分布面积较后两个分支大,并且在东部分支中可以发现一定的演化关系。结合古气候学和地质学信息,推测东部分支中现存的单倍型由冰期时气候变冷导致鄂报春快速迁移而形成,云南、四川分支中的单倍型可能是在各自的避难所中得以保存。同时本文也简要地探讨了鄂报春几个亚种之间的分类学关系。

欧报春(Primula vulgaris)不同花色与色素关系及花色遗传初步分析

为了掌握欧报春各花色遗传规律服务于良种生产,通过对欧报春各色花进行色素吸收光谱和薄层层析分析,进行不同花色杂交研究,分析了欧报春各色花所含色素类型及各花色遗传规律。结果显示欧报春群体含多种花色素,单株也可含有多种花色素,形成多变的粉色、红色及蓝色花。黄色深浅主要由类胡萝卜素含量决定。白色对粉色及黄色为隐性遗传,黄色、粉色为显性遗传并有数量遗传特征,黄色与粉色独立遗传。蓝色为多基因控制的隐性遗传,并具有数量遗传特征。

Optimization of the Ultrasonic-assisted Extraction Process for Total Flavonoids in Primula sikkimensis Hook.by Response Surface Methodology

[Objectives]To optimize the ultrasonic-assisted extraction process for total flavonoids in Primula sikkimensis Hook.[Methods]The effects of ethanol volume fraction,extraction time,extraction temperature,ultrasonic time and solid-to-liquid ratio on the yield of total flavonoids were investigated by single factor method,and the extraction process for total flavonoids in P.sikkimensis was optimized by Box-Behnken experimental design.[Results]The optimal extraction process for total flavonoids in P.sikkimensis was ethanol volume fraction 45%,extraction time 29 min,extraction temperature 80℃,solid-to-liquid 1∶40(g/mL)and ultrasonic time 10 min.Under such conditions,the average yield of total flavonoids in P.sikkimensis was 3.09%.[Conclusions]The optimized extraction process is simple and feasible,and is expected to provide a reference for the extraction of total flavonoids in P.sikkimensis.

Development and characterization of 43 microsatellite markers for the critically endangered primrose Primula reinii using MiSeq sequencing

Primula reinii(Primulaceae), a perennial herb belonging to the Primula section Reinii, occurs on wet,shaded rocky cliffs in the mountains of Japan. This threatened species comprises four varieties; these plants are very localized and rare in the wild. In this study, 43 microsatellite markers were developed using MiSeq sequencing to facilitate conservation genetics of these critically endangered primroses. We developed novel microsatellite markers for three varieties of P. reinii, and tested its polymorphism and genetic diversity using natural populations. These novel markers displayed relatively high polymorphism; the number of alleles and expected heterozygosities ranged from 2 to 6(mean = 3.2) and 0.13 to 0.82(mean = 0.45), respectively. All loci were in Hardye Weinberg equilibrium. These microsatellite markers will be powerful tools to assess P. reinii genetic diversity and develop effective conservation and management strategies.

鄂报春(Primula obconica Hance)一新亚种

A new subspecies, Primula obconica subsp. fujianensis, is described. The genus Primula is widely distributed throughout China, but it has never been previously recorded in Fujian and Jiangsu. Recently the authors collected a specimen of the genus from Fujian for the first time. It is obviously closely related to P. obconica, which is one of the most common species in China, ranging from western Yunnan and Sichuan through Guizhou, Hubei to Hunan and northern Guangdong, with its most easterly extension in southern Jiangxi. Morphologically, the Fujian plant is characterized by its much smaller stature and by the scapes being shorter than the leaf rosette, and in many ways it is more closely related to P. obconica subsp. parva, but can be easily distinguished by its smaller, white and homostylous flowers. As it is isolated both in distribution and reproduction with other subspecies of P. obconica, here it is described as a new subspecies.

Primula vulgaris extract induces cell cycle arrest and apoptosis in human cervix cancer cells

Primula vulgaris belongs to the genus Primula, members of which are frequently used in folk medicine.Various studies have investigated the cytotoxic effect of different Primula species, but there have been limited studies on the cytotoxic effect of P. vulgaris. The aim of this study was to investigate the cytotoxic effects, and possible mechanisms involved, of P. vulgaris flower extract on human cervical cancer(HeLa)cells. The cytotoxic effect of the extract on HeLa cells was revealed using the MTT assay. Mechanisms involved in the extract's cytotoxic effect were then investigated in terms of apoptosis, mitochondrial membrane potential, and the cell cycle, using fluorometric methods. P. vulgaris flower extract exhibited selective cytotoxic effects against HeLa cells by arresting their cell cycle at the S phase, and inducing the number of apoptotic cells compared to normal fibroblast cells by reducing mitochondrial membrane potential in a concentration-dependent manner. This is the first study to reveal the antiproliferative effect of P. vulgaris flower extract. Further studies are now needed to identify the cytotoxic molecules in the extract and their mechanisms.

四川省报春花科一新记录种——习水报春

本文报道了报春花科报春花属习水报春在四川省的地理新分布,简述了该种的形态特征、补充了形态图像,并与黔西报春和似绒毛报春在形态特征上进行了详细地比较。这一发现丰富了四川的生物多样性资料,也为日后的保护开发利用提供了基础。

Shortened anther-stigma distance reduces compatible pollination in two distylous Primula species

Aims Distyly has been regarded as an adaptation to improve compatible pollination between two floral morphs with reciprocal herkogamy.The hypothesis that the different positions of anthers and stigmas within flowers as well as their reciprocal position between morphs,reduce the probability of self pollination raised by Darwin has been rarely tested.in this study,we measured stigmatic pollen loads in response to reduced reciprocal herkogamy in two Primula species.Methods To see whether reciprocal herkogamy can increase compatible and/or reduce incompatible pollen deposition,thus promoting compat-ible pollination,we shortened the distance between anthers and stigmas within the flowers by changing the position of the corolla tube,to which the anthers were fused,i.e.reduced herkogamy in natural populations of Primula secundiflora and P.poissonii and quantified stigmatic pollen loads in the field over 2 years.Important Findingsin both species,stigmatic pollen loads were significantly higher in the long-styled(l-morph)than in the short-styled morph(s-morph)in both control and manipulated flowers,but percentage of compatible pollen in s-morph were higher.Flowers manipulated to halve the anther-stigma distance showed a similar pattern for 2 years:total pollen grain counts on stigmas did not differ significantly but compatible pollen grains in l-and s-morphs were significantly decreased in both species.The percentage of compatible pollen loads was decreased by 68.7%in P.secundiflora and 65.3%in P.poissonii in l-morphs,while it decreased by 30.6%and 2.9%in s-morphs,respectively.our manipulation of the relative position of anthers and stigmas in the two distylous species indicated that a lower degree of herkogamy reduced compatible but incompatible pollen transfer was likely to increase.The higher proportion of compatible pollen in the s-morph than in the l-morph in the two Primula species could be attributed to the accessibility of two-level sexual organs,floral orientations and pollinator behaviors.This is a first attempt to manipulate intraflower herkogamy for understanding adaptation of heterostyly,shedding insights into how the reciprocal herkogamy promotes compatible pollination.

报春石斛化学成分及药理作用研究进展

报春石斛(Dendrobium primulinumLindl.)是石斛属多年附生植物,外观艳丽,香气宜人,极具观赏价值。报春石斛的应用源于傣族传统医药业,具有消肿止痛、清热除湿、杀虫止痒、补土滋水等功效。目前,已确定报春石斛含有生物碱类、联苄类、菲类、黄酮类、苯丙素类等化学成分,具有多种药理作用,有助于抗癌、抗菌、抗炎、抗糖尿病、保肝。文章将对此展开综述,以期为后续研究提供理论支持。

四川报春花科2新记录种--散花报春和天竺葵报春

报道了四川省2种报春花科植物新记录物种,即散花报春(Primula effusa)和天竺葵报春(P.pelargoniifolia),该发现为四川省报春花属多样性研究和引种栽培提供了数据,凭证标本存放于中国科学院成都生物研究所标本馆(CDBI)。

Characterization of 24 microsatellite markers in Primula chungensis(Primulaceae),a distylous-homostylous species,using MiSeq sequencing

Primula chungensis is a species with considerable floral and mating-system variation,including distylous(outcrossing),homostylous(selfing) and mixed populations that contain both outcrossing and selfing forms.We isolated 24 microsatellite markers from P.chungensis using Illumina Mi Seq sequencing.Polymorphism and genetic diversity were then measured based on a sample of 24 individuals from a natural population in southern Tibet.All loci were polymorphic with the number of alleles per locus ranging from 2 to 4.The observed and expected heterozygosity ranged from 0 to 1 and 0.219 to 0.708,respectively.The microsatellite markers we have identified will serve as valuable tools for the investigation of the population genetic structure and phylogeography of P.chungensis and will inform models of the evolutionary history of mating systems in the species.

Fitness-related selection on floral traits in the distylous plant Primula poissonii (Primulaceae)

Aims Reproductive fitness of different floral phenotypes varies within and/or among populations.These variations are important to understand the process of natural selection and the evolution of floral traits.In this study,we focused on a distylous,self-incompat-ible species,Primula poissonii,to investigate fitness-related selec-tion on floral traits.Our aim was to determine how traits vary as targets of natural selection and whether morph-specific selection occurs.Methods This study was conducted at two sites(Yushuizhai at 2700 m and Haligu at 3200 m)in the Lijiang Alpine Botanical Garden,northwest-ern Yunnan,southwestern China.Insects visiting flowers of P.pois-sonii were observed,captured and identified.Randomly selected plants of long-and short-styled morphs were labeled.Five floral/inflorescence traits were measured including floral display,corolla width(CW),floral tube length(FTL),tube opening width(TOW)and floral scape height.Fruit and seed set were recorded.The total num-ber of seeds per individual plant(plant fitness)and seed production per capsule(flower fitness)were calculated.Multiple regression analyses were used to quantify selection gradients.Important Findings The frequencies of the two morphs did not deviate from the expected 1:1 ratio at both sites.Except for FTL,the four other traits did not dif-fer significantly between the long-and short-styled morphs.Floral scape height,floral display and FTL differed between two sites.The selection regimes differed between two morphs and between two sites.At the Yushuizhai site,linear selection for shorter floral tubes was stronger in the short-styled morph.However,nonlinear selec-tion on the floral display was stronger in the long-styled morph than selection on the short-styled morph.At the Haligu site,linear selec-tion for a smaller corolla was stronger in the long-styled morph.A morph-specific nonlinear selection on CW and floral display was also detected.Morph-specific selections were detected through the estimation of flower fitness only in Haligu population.In this site,morph-specific linear selection was also detected for CW and floral display.Morph-specific nonlinear selection on traits was detected only in CW.We found that butterflies and sphingid moths dominated at Yushuizhai,while long-tongued bees dominated at Haligu.The difference in pollinator fauna suggested that selection on floral tubes may be due to differences in pollinator assemblages.Overall,variation of floral and/or inflorescence traits in P.poissonii was probably driven by pollinator selection.Selection regime dif-ferences between two morphs,in part,due the inter-morph diver-gences of sexual functions in distylous plant.

毛报春(Primula×pubescens)腋芽再生组织培养体系的建立

以毛报春(Primula×pubescens)无菌腋芽为外植体,分析不同浓度激素配比对愈伤组织诱导和分化以及不定芽增殖和生根的影响,筛选出不同阶段的最适培养基,优化毛报春的组织培养再生体系。结果表明,毛报春腋芽愈伤组织诱导及分化的最适培养基为MS+0.2 mg·L^(-1)NAA+1.0 mg·L^(-1)6-BA,诱导率达84%,出芽率达67%;不定芽增殖最适培养基为MS+0.5 mg·L^(-1)NAA+0.2 mg·L^(-1)6-BA,增殖率可达67%,苗绿且健壮;MS+0.2 mg·L^(-1)NAA培养基最有利于组培苗的生根及伸长,平均单株生根数为9条,生根率高达70%。该研究建立了毛报春的组织培养再生体系,可为报春属其它植物的遗传研究及种质创新提供参考。

Effects of Population Size on Reproductive Success of the Endangered and Endemic Species Primula merrilliana

The reproductive ability related to the population size of the endangered and endemic species Primula merrilliana Schltr. was investigated. In 26 populations observed, only four contain more than 500 flowering individuals, whereas most of them (53.8%) consist of less than 100 flowering individuals. Though the ratio of pin and thrum plants keeps its balance at 1:1 for all populations, the frequency of pin and thrum flowers was significantly biased in most small populations. Population size strongly affected reproductive success; plants in small populations produced significantly fewer flowers and fruits per plant and fewer seeds per fruit, and therefore fewer seeds per plant. The floral density was another important factor that influenced the reproductive success of P. merrilliana, because four main reproductive success parameters (fruits per plant, seeds per fruit, seeds per plant, and the proportion of flowers setting fruit) were all positively correlated with floral density. The size of plants and the number of leaves per plant (measure of habitat quality) were not influenced by the variation of population size, suggesting that the reduced fecundity in small populations may not be a consequence of lower habitat quality. Inbreeding depression and pollen limitation as a result of less attractiveness in small populations are therefore likely explanations for these patterns.

珍稀濒危植物陕西羽叶报春的开花及传粉生物学研究

陕西羽叶报春为中国珍稀濒危植物,研究其开花物候、传粉者及访花行为、花粉活力、柱头可授性、繁育系统,以明确陕西羽叶报春的传粉特性及制濒机制。以勉县陕西羽叶报春野生居群为研究对象,通过野外观察和人工授粉实验首次对陕西羽叶报春的传粉生物学进行研究。结果表明:(1)陕西羽叶报春种群花期为2-4月中旬,单花花期一般为14~17 d;单花开花进程中花部形态有6个明显发育变化:全部被萼片包裹、花苞伸出并为深粉色、花朵打开为淡粉色、花冠为白色、花朵枯萎、花冠脱落花萼宿存。(2)陕西羽叶报春为异花授粉植物,蜜蜂属为其主要传粉者。(3)柱头在花粉未开裂前就具有可授性,可授性逐渐增强;花药在花瓣打开后的第3天才开始开裂、散粉,在第4天活力最高,为98.18%,此后逐渐减弱。(4)繁育系统检测显示,长、短柱型花的花粉/胚珠比分别为902.26和831.48,杂交指数为4;套袋实验显示,陕西羽叶报春异型交配可育,同型交配不可育。由此表明,陕西羽叶报春的繁育系统为异交型,自交不亲和,需要传粉者。综上所述,陕西羽叶报春传粉者种类少、自交不亲和、胚珠败育现象可能是导致陕西羽叶报春濒危的重要原因。