头花马先蒿和管花马先蒿的化学成分

从头花马先蒿全草中发现13个化合物,经波谱分析鉴定为：（±）-松脂素（1）、金合欢素（2）、木犀草素（3）、7．去氧栀子新苷（4）、yuheinoside（5）、euphroside（6）、mussaenosidic acid（7）、莫桑苷（8）、桃叶珊瑚苷（9）、角胡麻苷（10）、异角胡麻苷（11）、贞桐苷A（12）和开德苷元（13）。从管花马先蒿全草中发现4个化合物,经鉴定为：（＋）．dehydrovomifoliol（14）、催吐萝芙木醇（15）、ω-hydroxypropioguaiacone（16）和3-hydroxy-1-（4-hydroxy-3,5-dimethoxyphenyl）-1-propanone（17）。所有化合物均为首次在该种中发现。     

A new synonym of Pedicularis cyathophylla (Orobanchaceae)

Pedicularis cyathophylla Franchet in Bull.Soc.Bot.France 47:25.1900.Type:China.Sichuan:Kangding,Zheduoshan Mountains (Ta-tsien-kou & Tche-to-chan),J.A.Soulié 218 (holotype,P00160827,P!).Pedicularis xiangchengensis H.P.Yang in Acta Phytotax.Sin.28:137.f.3:3.1990,syn.nov.Type:China.Sichuan:Xiangcheng,Rewu,on mountain slopes,alt.4100-4300 m,1981-08-06,Qing-Zang Exped.3829(holotype,00032964,PE!;isotypes,KUN!,CDBI!).     

Floral nectary morphology and evolution in Pedicularis (Orobanchaceae)

Intricate associations between floral morphology and pollinator foraging behaviour are common. In this context, the presence and form of floral nectaries can play a crucial role in driving floral evolution and diversity in flowering plants. However, the reconstruction of the ancestral state of nectary form is often hampered by a lack of anatomical studies and well‐resolved phylogenetic trees. Here, we studied 39 differentially pollinated Pedicularis spp., a genus with pronounced interspecific variation in colour, shape and size of the corolla. Anatomical and scanning electron microscopy observations revealed two nectary forms [bulged (N = 27) or elongated (N = 5)] or the absence of nectaries (N = 7). In a phylogenetic context, our data suggest that: (1) the bulged nectary should be the ancestral state; (2) nectaries were independently lost in some beaked species; and (3) elongated nectaries evolved independently in some clades of beakless species. Phylogenetic path analysis showed that nectary presence is indirectly correlated with beak length/pollinator behaviour through an intermediate factor, nectar production. No significant correlation was found between nectary type and nectar production, beak length or pollinator behaviour. Some beaked species had nectary structures, although they did not produce nectar. The nectary in beaked species may be a vestigial structure retained during a recent rapid radiation of Pedicularis, especially in the Himalaya–Hengduan Mountains of south‐western China. © 2015 The Linnean Society of London, Botanical Journal of the Linnean Society, 2015, 178 , 592–607.     

Floral colour change in Pedicularis monbeigiana (Orobanchaceae)

We examined the effects of the retention of colour-changed flowers on long- and short-distance attractiveness of bumblebees and the likelihood of successive flower visits by bumblebees in Pedicularis monbeigiana. The lower lip changed colour with age from white to purple. Hand geitonogamous pollination significantly reduced seed production. No pollen limitation occurred in this species. Purple-phase flowers contributed minimally to pollinator attractiveness at long distance. The combination of less reproductive flowers with a lower amount of reward and floral colour change enabled plants to direct pollinators to reproductive, highly rewarding white flowers at close range. A high percentage of purple-phase flowers in an inflorescence was associated with a marked reduction in the frequency of successive flower visits to individual plants. We suggest floral colour change in P. monbeigiana may serve as a mechanism for enhancing inter-individual pollen transfer and reducing intra-individual pollen transfer.     

Pollen-Ovule Ratio and Gamete Investment in Pedicularis （Orobanchaceae）

The Pedicularis species provides ideal materials to study floral evolution because of their substantial flower variation based on a narrow genetic basis, even though they are almost exclusively pollinated by bumblebee. These traits allow us to detect the evolutionary trends of floral parameters without considering genetic background and the difference of pollination vectors. The pollen-ovule ratio is widely used to estimate the pattern of resource investment in two sexual functions in flowering plants. Forty species representing all of the corolla types in Pedicularls were used to study pollen-ovule ratio, gamete investment, and their correlations. Results show that pollen-ovule ratio does not differ among both different corolla types and taxonomic groups. It is therefore suggested that pollen-ovule ratio should be a parallel evolution. The correlations between pollen-ovule ratio and pollen size （-）, and ovule size （＋） can be successfully explained in terms of sex allocation theory. The biological significance of such relationships was also discussed. Additionally, we analyzed the pattern of resource investment into female gamete, which has been somewhat neglected, and found that plants have different patterns of gamete investment between the two sexual functions.     

马先蒿属花冠无喙类的花器官发生

对花冠无喙类密穗马先蒿(Pedicularis densispica)和大王马先蒿(P．rex)的花器官电镜扫描发现,两种不同花冠型(无齿和具齿)的马先蒿花部器官发生和发育初期十分相似,表现为明显的单轴对称。2个萼片原基首先发生于花顶的近轴侧位,然后沿花顶边缘向远轴端发育形成--马蹄形结构。密穗马先蒿在近轴中部又出现1枚萼片原基,随后马蹄形结构分化出4枚萼片,并与近轴中部的原基愈合后构成5齿萼片;而大王马先蒿的2齿萼片直接由马蹄形结构发育而成。5枚独立的花瓣原基随后发生,但发育相对滞后;除近轴中部位置1枚空缺外,4枚雄蕊原基与花瓣原基位置呈交互发生;2个心皮原基同时在拱形花顶的近轴和远轴端发生,剩余的花顶形成中间的隔膜,并与2个心皮形成中轴胎座。对马先蒿与金鱼草(Antirrhinum majus)和毛地黄(Digitalis purpurea)花器官发生和发育初期的特征进行了比较,讨论了马先蒿属花冠对称性变化的意义。     

Pollen size-number trade-off and pollen-pistil relationships in Pedicularis (Orobanchaceae)

Current patterns of floral design in Pedicularis must have undergone an evolutionary process of interacting among components of floral traits, and then formed internal relationships among these traits. To detect such correlations, which may provide insight to understand flower evolution, 40 Pedicularis species representing all corolla types of the genus were studied. Results show that, interspecifically, pollen size correlates negatively with pollen number, but positively with pistil length. This suggests that plants evolve an optimal pollen size, which balances the advantages of large pollen size for gametophytic competition against the fecundity disadvantages of fewer pollen grains. In contrast to sex allocation theory, this study does not find a trade-off, but an interspecific positive correlation between pollen and ovule number. This is consistent with the hypothesis that genetic variation for resource acquisition may in part be responsible for the lack of negative correlation between male and female function.This work was supported by the State Key Basic Research and Development Plan, China (Grant No. G2000046804) to YHG. The authors would like to thank Peter K. Endress and two anonymous reviewers for providing critical comments and helpful suggestions, Qing-Feng Wang, Jing-Yuan Wang and Jin-Ming Chen for their helpful suggestions. Shi-Guo Sun, Jing Xia, and Qian Yu are thanked for their assistance in both the field work and laboratory phases of the project.     

Seed morphological diversity of Pedicularis (Orobanchaceae) and its taxonomic significance

Mature seeds of 109 species of Pedicularis representing 13 groups according to Tsoong’s infra-generic classification were investigated using light microscopy (LM) and scanning electron microscopy (SEM). The largest seeds occurred in P. superba (4.82 ± 0.32 mm × 2.18 ± 0.18 mm) and the smallest in P. crenata (0.95 ± 0.16 mm × 0.72 ± 0.08 mm). The individual seed weight ranged from 0.20 to 9.52 mg. There were two types of primary ornamentation, i.e. reticulate and undulate. Reticulate seeds were found in all examined species except for P. pantlingii and P. confluens. The reticulate seeds were divided into three subtypes, i.e. cristate-reticulate, regular-reticulate and membranous-reticulate. Among them, the cristate-reticulate and the undulate primary ornamentation are firstly reported. The inner tangential wall ornamentation was categorized into granulate, ruminate or striate. Seed characters seem to support the transfer of Pedicularis to Orobanchaceae from traditional Scrophulariaceae. Seed length, individual seed weight, primary ornamentation and inner tangential wall ornamentation were considered as potentially useful characters for infra-generic classification. Non-metric multidimensional scaling (NMDS) analyses indicated a clustering of some species in group 1 (Sceptrum) and all species in group 6 (Cyathophora), respectively. However, seed data were not highly congruent with Tsoong’s classification for delimitation of existing groups. Some characters, such as seed shape, primary ornamentation, inner tangential wall ornamentation and epidermal cell shape could be applied to identification of some species in this genus.     

马先蒿属植物花冠分化与繁殖适应的研究进展

结合已有的研究报道和作者近年来的工作, 对马先蒿属(Pedicularis)植物的花冠多样化成因与繁殖适应特性进行了总结和探讨。通过对该属4种进化花冠型的花器官发生和分化的研究发现, 花部各器官在发生和发育初期基本一致, 后期上唇形态的分化是导致成熟花形态结构产生较大差异的重要阶段。孢粉学研究认为, 花冠类型与花粉萌发孔类型之间具有显著相关性; 萌发沟的演化可能与繁殖适应有一定的关系。分子系统学研究表明, 多样化的花冠类型在不同的谱系内经过若干次的独立进化而表现出了高度的平行演化(p ara l lel ism )。传粉生物学研究证实, 该属植物花冠多样化与其主要传粉者熊蜂属(Bombus)昆虫的传粉行为存在较为密切的关系。具有相同(似)花冠类型的马先蒿可能被同种或不同种的熊蜂以相同的方式访问, 但在花粉落置位置上存在显著差异, 这可能有助于同域分布重叠的物种间在生殖上的机械隔离, 而花冠的分化在一定程度上促进了新的物种形成。     

马先蒿属植物花冠分化与繁殖适应的研究进展

结合已有的研究报道和作者近年来的工作,对马先蒿属（Pedicularis）植物的花冠多样化成因与繁殖适应特性进行了总结和探讨。通过对该属4种进化花冠型的花器官发生和分化的研究发现,花部各器官在发生和发育初期基本一致,后期上唇形态的分化是导致成熟花形态结构产生较大差异的重要阶段。孢粉学研究认为,花冠类型与花粉萌发孔类型之间具有显著相关性;萌发沟的演化可能与繁殖适应有一定的关系。分子系统学研究表明,多样化的花冠类型在不同的谱系内经过若干次的独立进化而表现出了高度的平行演化（parallelism）。传粉生物学研究证实,该属植物花冠多样化与其主要传粉者熊蜂属（Bombus）昆虫的传粉行为存在较为密切的关系。具有相同（似）花冠类型的马先蒿可能被同种或不同种的熊蜂以相同的方式访问,但在花粉落置位置上存在显著差异,这可能有助于同域分布重叠的物种间在生殖上的机械隔离,而花冠的分化在一定程度上促进了新的物种形成。     

Corolla wilting facilitates delayed autonomous self-pollination in Pedicularis dunniana (Orobanchaceae)

Structural changes associated with corolla wilting may serve as a mechanism for effecting self-pollination. Low pollinator visitation, high seed production and a corolla that persists after anthesis indicates that Pedicularis dunniana is autogamous. Delayed autonomous self-pollination is facilitated by corolla wilting. Wilting of the upper lip (galea) brought the pollen laden anthers into contact with the stigma resulting in the deposition of self pollen on the stigma. The seed set of flowers either emasculated, or with restrained galeae thus preventing anthers brushing against the stigma, was significantly lower than that of open-pollinated flowers. This demonstrates that autogamy occurs in this species through corolla wilting. Germination experiments indicated that outcross seedlings were more vigorous than selfed seedlings as a result of inbreeding depression. It is likely that autogamy provides reproductive assurance for P. dunniana under conditions of pollinator scarcity.     

Honeybees enhance reproduction without affecting the outcrossing rate in endemic Pedicularis densispica (Orobanchaceae)

There has been substantial debate in recent years surrounding the impact of introduced honeybees on native biota. This study reports on an investigation of Pedicularis densispica, a subalpine annual herb endemic to Southwest China, in an attempt to determine the impact of introduced domestic honeybees on pollen dispersal and thus on their reproductive success and mating system. Honeybees were introduced into the study site in 2004, and a sudden seasonal pollinator shift from bumblebees to honeybees was observed. Intra- and inter-plant visits by different pollinators were recorded in the field in 2003 and 2004. Fruit and seed sets prior to and after the pollinator shift were measured. Experimental pollinations were performed to characterize the breeding system. Outcrossing rates at the seed stage were estimated for both years using RAPD markers. Our results indicated that honeybees foraged between plants more frequently than bumblebees did. Our results also revealed that the introduction of honeybees significantly enhanced reproductive success. However, no significant difference was detected between the outcrossing rates due to bumblebee and honeybee pollination. P. densispica was almost completely outcrossing ( T(m) = 0.956 and 0.967, respectively in 2003 and 2004) but partially self-compatible. This study presents the first report of the outcrossing rate in the genus pedicularis and reveals a limited influence of pollination on the mating system in P. densispica. The pollinator shift did not reduce reproductive success of the plants and honeybees may be used to augment pollinator services for nectariferous P. densispica.     

Growth, Resource Allocation and Haustorial Biology of the Root Hemiparasite Olax phyllanthi (Olacaceae)

Olax phyllanthi was found to parasitize a wide range of taxain the native habitat in coastal heath, South-West Australia.All major life and growth forms were regularly exploited, includingmost woody dicotyledons (except members of the Myrtaceae), afew monocotyledons and cohabiting root hemiparasites. Initiationof haustoria occurred mostly in autumn (southern hemisphere),with some senescing the following summer, and others survivinguntil replaced by a new generation of haustoria the followingautumn. Seedlings increased in dry matter and contents of N,P, K, Mg and Ca during preparasitic development, but did notsurvive beyond 6 months if failing to establish haustoria ona suitable host. Plant dry matter, mineral content and haustorialnumber increased exponentially during subsequent parasitic development.Mean shoot:root d. wt ratios of 1st to 3rd-year plants lay withinthe range 1.2–1.4. Root systems were laterally extensiveand restricted to the top 40 cm of rooting substrate. Mean totalroot lengths of 2nd- and 3rd-year plants were 7.1m (n = 5) and60.9 m (n = 5), respectively. Haustoria comprised 0.7–3.5%of plant d. wt, with a mean of 7.9 haustoria (n = 10, 2nd- and3rd-year plants) per metre of root length. Comparisons of mineralconcentrations in dry matter of O. phyllanthi and of a rangeof commonly parasitized hosts showed the parasite to be muchricher on average than its hosts in K, P, and to a lesser extentin N, but not noticeably different in Mg and Ca. Olax, root hemiparasite, mineral nutrition, haustoria, resource allocation     

Pollen morphology in relation to floral types and pollination syndromes in Pedicularis (Orobanchaceae)

Floral diversification in the genus Pedicularis (Orobanchaceae) is remarkable among flowering plants. In this genus, floral morphology and pollinator behavior are closely co-adaptive. In the current paper, pollen grains of 23 representative species of Pedicularis mainly from North America, with two species from Japan and two species from China, whose pollination ecology was previously studied, were examined using light microscopy and scanning electron microscopy. Two pollen aperture types and three kinds of exine ornamentation were recognized among these species. In addition, pollen data from previous and the current studies of Pedicularis were integrated and analyzed, together with some pollination characters. There was a significant association between pollen aperture types and corolla types, as well as between pollination syndromes and corolla types. However, there was no association of exine ornamentations with corolla types. The relationships and evolution of this genus were discussed with regards to pollen morphology, corolla types and pollination syndromes.     

Pectolytic Activity by the Haustorium of the Parasitic PlantOrobancheL. (Orobanchaceae) in Host Roots

The possible involvement of enzymes in the penetration of intrusivecells of the parasitic angiospermOrobancheinto host root tissueswas studied using cytochemical and immunocytochemical methods.Pectin methyl esterase (PME) was detected, with specific antibodies,in the cytoplasm and cell walls ofOrobancheintrusive cells andin adjacent host apoplast. Depletion and chemical changes ofpectins in host cell walls were shown by histochemical stainingwith PATAg, which detects carbohydrates that are sensitive toperiodic acid, especially pectins, and with the monoclonal antibodiesJIM 5 and JIM 7 that label pectins with low and high rates ofesterification, respectively. Galacturonic sequences with lowrates of esterification were more abundant in host cell wallsadjacent to the parasite, which is consistent with pectin de-methylationby PME release from the parasite. Pectins were absent in middlelamellae and in host cell walls neighbouring mature intrusivecells of the parasite, consistent with further degradation ofpectins by other enzymes. These results provide the first directevidence for the presence and activity of a pectolytic enzymein the infection zone of the haustorium of a parasitic angiosperminsitu.Copyright 1998 Annals of Botany Company Broomrape;Orobanche; parasitic weed; haustorium; pectin methyl esterase; pectin; cell wall.     

Host Associations of the Introduced Annual Root Hemiparasite Parentucellia viscosa in Agricultural and Bushland Settings in Western Australia

Populations of the introduced annual root hemiparasite, Parentucelliaviscosa (L.) Caruel (Scrophulariaceae), were examined in habitatswhere introduced, mixed introduced plus native, or solely nativespecies provided potential hosts. Presence of haustoria on hostroots confirmed parasitism of 17 introduced and ten native taxaacross the sites investigated. Paired plots, one with all hostsremoved early in the season, the other left intact, showed overallincreases in shoot dry matter of the parasites over 3 monthsof growth. Results indicated a substantial growth benefit tothe parasite from continued access to introduced and indigenousplant species, although the parasite continued growing to alimited extent after removal of hosts. Carbon isotope discriminationvalues (     

Fast and abundant in vitro spontaneous haustorium formation in root hemiparasitic plant Pedicularis kansuensis Maxim. (Orobanchaceae)

Haustorium formation is the characteristic feature of all parasitic plants and a vital process for successful parasitism. Previous investigations on haustorium initiation and development are constricted to induced processes by host-derived signals or synthetic analogs. Spontaneous haustorium formation in the absence of host signals, a process representing an early stage in the evolution of parasitic plants, remains largely unexplored. Lack of fast and frequent formation of spontaneous haustoria greatly hinders full understanding of haustorium formation in root hemiparasites. In this study, seedlings of Pedicularis kansuensis Maxim., a facultative root hemiparasitic species in Orobanchaceae observed to produce many spontaneous haustoria, were grown in autoclaved water agar in the absence of any known haustoriuminducing stimulants. We aimed to test the temporal and developmental pattern of spontaneous haustorium formation. Also, effects of sucrose supply and root contact on spontaneous haustorium formation were tested. Spontaneous haustoria were observed starting from six days after germination, much earlier than previously reported root hemiparasites. A majority of the spontaneous haustoria formed on lateral roots. Percentage of seedlings with spontaneous haustoria was 28.8% when grown on water agar plates, with a mean of four haustoria per seedling two weeks after germination. Haustorium formation by seedlings grown in water agar amended with 2% sucrose was more than twice of those without sucrose amendment. Singly grown seedlings were able to develop spontaneous haustoria at similar levels as those grown with another conspecific seedling. In view of the fast and abundant formation of spontaneous haustoria, P. kansuensis may be developed as an excellent experimental system in future investigations for unraveling endogenous regulation of haustorium initiation and development in root hemiparasitic plants.     

Unexpected high divergence in nrDNA ITS and extensive parallelism in floral morphology of Pedicularis (Orobanchaceae)

Internal transcribed spacer (ITS) region sequences of nuclear ribosomal DNA (nrDNA) were used in the phylogenetic reconstruction of Pedicularis, a genus with strong adaptive radiation. Forty-two species representing 12 greges of the genus were all, except P. resupinata, sampled from the Hengduan Mountain region, China. A high level of ITS sequence variation was found among the species distributed in such a small geographical area, which had been rarely reported in other groups. The great ITS divergence in Pedicularis could be explained by a relatively ancient origin and diversification of the genus followed by migration of different floristic components into the Hengduan Mountains, or accelerated rates of molecular evolution in parasitic lineages of Orobanchaceae. In the present ITS phylogeny, almost all the main clades are not consistent with the high hierarchical taxa within Pedicularis, which implies that significant parallel evolution occurred in floral morphology of the genus, and that undue attention has been paid to corolla characters in the intragenus classifications.We thank the two anonymous reviewers for valuable comments and suggestions; Professor Guo You-Hao, Drs. Tan Dun-Yan, Luo Yi-Bo, Bao Ying, Luo Yan, Mr. Mao Jian-Feng and Mr. Qin Wei for their great help in the field collection; Drs. Song Bao-Hua, Wei Xiao-Xin and Miss Sun Ying-Xue for their help in the laboratory work. This study was supported by State Key Basic Research and Development Plan (Grant No. G2000046804) and the National Natural Science Foundation of China (Grant No. 30121003).     

Unexpectedly high outcrossing rate in both dense and sparse patches in self-compatible Pedicularis rex (Orobanchaceae)

Pedicularis rex, a self-compatible alpine herb, depends exclusively on pollinators for reproduction. Previous study has revealed that 74.7 % of genetic variability occurs among populations in Yunnan, indicating a mixed mating system for the species. The large floral display also indicates high potential geitonogamy. Therefore, plant density and display size may be important factors influencing the realized mating system of P. rex. In the study reported here we explored the effects of plant density and floral display size on pollinator movements and thus outcrossing rate in two patches of in P. rex at low and high plant densities. We examined foraging behavior of bumblebees, measured reproductive outputs under hand and natural pollination, and estimated the outcrossing rates using RAPD markers. Floral display size was significantly larger and bumblebees visited significantly more flowers in sequence on individual plants in the sparse patch than in the dense patch. We found a significant positive correlation between floral display size and successive visits in the sparse patch, but not in the dense patch. Regression analysis revealed a significant inverse correlation between fruit production and flower production per plant with a theoretical maximum of 63 fruits per plant, but no relationship between seed production and flower production per plant. Contrary to previous studies on genetic structure, however, we found an unexpectedly high outcrossing rate in both the sparse and the dense patch (t _m = 1.151 and 0.924, respectively). Resource limitation may intensify seed competition and result in selected seed abortion, which may largely explain the unexpectedly high outcrossing rate.