虎耳草属异叶虎耳草复合群的数量分类学研究

山羊臭组（Saxifraga sect. Ciliatae Haworth）隶属于虎耳草属（Saxifraga Tourn. ex L.）,具有极高的物种丰富度,但山羊臭组内部的系统发育关系一直都未能很好解决。唐古拉亚组（S. subsect. Hirculoideae Engl.）是山羊臭组中最大的亚组,主要分布于青藏高原及其周边地区。根据形态特征,可将唐古拉亚组划分为3个复合群,其中叶背边缘有突起叶脉的物种被归入异叶虎耳草复合群（S. diversifolia complex）。本研究通过标本查阅,选取异叶虎耳草复合群24个物种、2变种共657份标本,对其24个定性性状信息进行主成分分析和聚类分析;获取443条地理分布信息,构建复合群的分布式样。特征值大于1的前7个主成分的累计贡献率较低,仅为67.748%。“茎生叶是否具柄”“中下部茎生叶形态”“花序类型”“中下部茎生叶大小”“萼片脉纹于先端汇合”“叶基心形”等性状对前3个主成分的贡献值大,可作为异叶虎耳草复合群物种分类与鉴定的关键性状。基于形态聚类结果和地理分布式样,可将异叶虎耳草复合群划分为3个分支：喜马拉雅山南坡分支、环四川盆地山区分支以及横断山分支（包含一个广布种S. egregia）。主成分分析结果支持将异叶虎耳草复合群划分为3个分支。     

Influence of nitrogen supply and growth irradiance on photoinhibition and recovery in Heuchera americana (Saxifragaceae)

Prior work demonstrated that Heuchera americana, an evergreen herb inhabiting the deciduous forest understory in the southeastern United States, has a 3-4-fold greater photosynthetic capacity under the low-temperature, strong-light, open canopies of winter compared to the high-temperature, weak-light, closed canopies of summer. Moreover, despite the reductions in soil nitrogen, the chilling temperatures, and the increased quantum flux associated with winter, chronic photoinhibition was not observed in this species at this time of the year. We were interested in the photosynthetic acclimation and photoinhibition characteristics of this species when grown under contrasting light and nitrogen regimes. Newly expanded shade-acclimated leaves of forest-grown plants exposed to strong light varying in intensity and duration at 25°C showed a reduction in F_v/F_m (the ratio of variable to maximum room temperature chlorophyll fluorescence measured after dark adaptation), which was correlated with a decline in ø_a (the intrinsic quantum yield of CO₂-saturated O₂ evolution on an absorbed light basis). Plants grown in the glasshouse under contrasting light (high and low light; HL and LL, respectively) and nitrogen supply (high and low nitrogen; HN and LN, respectively) regimes showed that photosynthetic acclimation to HL was impaired in LN regimes. The HL-LN plants also had the lowest values of F_v/F_m and of ø on both incident and absorbed light bases and had 50% less chlorophyll (per unit area) compared to plants from other growth regimes. Controlled exposure to bright light at low temperatures (2-3°C) for 3 h resulted in a sharp decrease in F_v/F_m (and rise in F_o, the minimum fluorescence yield) in all plants. Shade-grown plants from both N regimes were highly susceptible to chronic photoinhibition, as indicated by a greater reduction in F_v/F_m and incomplete recovery after 18 h in weak light at 25°C. The HL-HN plants were the least susceptible to chronic photoinhibition, having the smallest decrease in F_v/F_m with near full recovery within 6 h. The decline in Fv/Fm in HL-LN plants was comparable to that of shade-acclimated plants, but recovered fully within 6 h. Low-N plants from both light regimes displayed greater increases in F_o which did not return to pretreatment levels after 18 h of recovery. These studies indicate that HL-LN plants were sensitive to chronic photoinhibition and, at the same time, had a high capacity for dynamic photoinhibition. Experimental garden studies showed that H. americana grown in an open field in summer were photoinhibited and did not fully recover overnight or during extended periods of weak light. These results are discussed in relation to the photosynthetic acclimation of H. americana under natural conditions.     

Pollen tube penetration and fertilization in Lilium longiflorum (Liliaceae)

The ultrastructure of the embryo sac, nucellus, and parts of the micropyle of Lilium longiflorum were studied both before and after pollen tube penetration to examine the interactions between ovule and pollen tube, using transmission electron microscopy and light microscopy. Before pollen tube penetration the egg cell and two synergids are similar. No filiform apparatus was detected and no synergid degeneration occurs prior to pollen tube penetration. The polar nuclei do not fuse until fertilization. No differences in embryo sac ultrastructure were detected between pollinated ovules unpenetrated by pollen tubes and unpollinated flowers of a comparable age. Shortly after the discharge of the pollen tube two enucleated cytoplasmic bodies with different ribosome densities were observed in the degenerated cytoplasm. These structures border both on the central cell and the egg cell as well as each other and are interpreted as remains of sperm cytoplasm after transmission of sperm nuclei. In the central cell both the sperm nucleus and the polar nuclei are associated with endoplasmic reticulum (ER). ER is thought to be a transport mechanism to achieve contact between the haploid polar nuclei and the sperm nucleus. In the egg cell sperm nucleus alignment is not visibly achieved by ER. The persistent cells of the egg apparatus and the central cell appear to become more metabolically active after pollen tube penetration. Pollen tube penetration already occurs despite the absence of a filiform apparatus and a low level of differences between the cells of the egg apparatus.     

Effect on in Vitro Pollen Growth of an Isolated Style Glycoprotein Associated with Self-Incompatibility in Nicotiana alata

The effect on in vitro pollen tube growth of an isolated style glycoprotein (S₂-glycoprotein) associated with self-incompatibility in Nicotiana alata was investigated. Tube growth of pollen bearing the S₂-allele was inhibited, but tube growth of pollen bearing other alleles was not affected. Inhibition showed a dose response effect. The percentage of pollen grains that germinated was not significantly affected by the S₂-glycoprotein. Growth of S₂-pollen in the presence of the S₂-glycoprotein resulted in increased binding to the pollen of monoclonal antibody (PCBC3) which has a primary specificity for α-l-arabinofuranosyl residues. Growth of pollen bearing other alleles in the presence of the glycoprotein resulted in no increased binding of the antibody.     

Gynodioecy inSaxifraga granulata L. (Saxifragaceae)

The occurrence of gynodioecy in two populations in northern England of the normally hermaphroditeSaxifraga granulata is reported. Female plants have aborted stamens, and smaller petals than hermaphrodites. At Staindrop, County Durham, an estimated 23% of the flowering stems were female; at Macclesfield, Cheshire, 4% were female. The inheritance of male sterility is not simple, and probably involves at least one cytoplasmic and two nuclear genes. The secondary sexual characteristics, hermaphrodite-predominant sex ratios, and complex inheritance of male sterility, are typical of gynodioecious populations.     

Pollen Cryopreservation and Pollen Protoplast Isolation in Brassica carnpestris var.purpurea

The authors have investigated the factors affecting pollen cryopreservation in Brassica campestris var. purpurea, such as pollen development stages cryoprotectant and the process of freezing. A suitable procedure was established as follows :Pollen grains suspended in B5 medium containing 10X DMSO and 1SM sucrose were frozen by a three- –1℃/min – 1 ℃/min step method(0℃———→–10 ℃ ,standing for 15 min———→–40 ℃ , standing for 1 hr→liquid nitrogen)and later thawed in 40℃ water bath. During a period of 60, 90 days′preservation, the relative survival percentage of mature (at the day of anthesis)and nearly mature(2 days before anthesis, trinucleate stage)pollens maintained at ca. 91% that of young pollens(7-8 days before anthesis, late uninucleate stage to early binucleate stage)slightly declined from the original 91.6% to 84. 3%. Culture. experiment showed that the cryopreserved young pollen could be induced to cell division just as well as the fresh pollen. The method of isolating protoplasts from fresh mature pollen developed previously was improved and simplified. As a result, protoplasts were isolated more conveniently from mature pollen and young pollen for the first time. The protoplasts from cryopreserved mature and young pollen could be obtained as well with an isolation rate of 77.4% and 35.9% respectively. However, for isolation of protoplasts from preserved young pollen, an incubation in NLN medium at 35℃ after thawing was necessary.     

Pollination and Reproductive Biology of Rauvolfia grandiflora (Apocynaceae): Secondary Pollen Presentation, Herkogamy and Self-Incompatibility

Abstract: The pollination and reproductive biology of Rauvolfia grandiflora were studied in natural populations in the forest of the Recife Botanical Garden. R. grandiflora is a shrubby species (2 to 5 m), the flowers are hermaphrodite and salverform. The corolla tube is white and the five free lobes of the corolla are flushed with violet. The five stamens are attached to the corolla tube; the anthers are introrse and form a cone situated immediately above the stigmatic head. There is a space between the anthers and the stigmatic head, where the pollen is deposited. The stigmatic head has three functional regions: a) an upper sterile region; b) a median region that produces a sticky substance and c) a lower receptive region, which is located beneath a basal collar. The secondary pollen presentation and the herkogamy mechanisms are discussed. Sugar concentration in nectar was, on average, 31.7%. The pollen viability was 97.4%. R. grandiflora is melittophilous and in the study area was found to be pollinated by only one species of long-tongued bee, Exaerete smaragdina. R. grandiflora was found to be self-incompatible, and the percentage of fruit set under natural conditions was low.     

Pollen tube growth and self-incompatibility in three Ziziphus species (Rhamnaceae)

Self-incompatibility and synchronous protandrous dichogamy have previously been reported in Ziziphus species. In this work, we conducted a comparative analysis of fluorescence microscopy observations of pollen tube growth following controlled cross pollinations of emasculated flowers and self-pollinations of non-emasculated flowers in three Ziziphus species, Z. jujuba, Z. mauritiana and Z. spina-christi, with the aim to determine the type of the self-incompatibility system of each species. In addition, to test whether autonomous self-pollination, parthenocarpy or agamospermy occurs, flowers were emasculated (or not) and covered. Fruit set and seed viability were monitored. The presence of binucleate pollen grains and the cessation of pollen tube growth in the style suggest that the self-incompatibility system operating in the studied Ziziphus species is gametophytically controlled. Controlled self-pollination in Z. mauritiana resulted in fruits that dropped off before maturation, whereas in Z. spina-christi the flowers dropped off one or two days after pollination. Following controlled self-pollination, small fruits lacking viable seeds were obtained in Z. jujuba, probably due to the stimulus provided by pollination (stimulative parthenocarpy). In the cultivar Tamar of Z. jujuba, the relatively high percentage of seedless fruits obtained in emasculated bagged flowers without hand pollination suggests that this cultivar can set seedless fruits without any pollination stimulus.     

Pollen tube expression of pseudo-self-compatibility (PSC) inPetunia hybrida

Summary AnS _1.1 self-incompatible (SI) petunia plant which showed atypical seed set was found in an I₇ population. This plant showed a strong SI reaction when selfed but produced varying amounts of seed when used as the seed parent in crosses with unrelated individuals homozygous for the sameS allele. Reciprocal crosses yielded no seed indicating that the reaction was a stylar response. Self seed obtained by high temperature treatments produced 18 plants, all of which exhibited the parental characteristics, the ability to reject self pollen but accept, to varying degrees, pollen bearing the sameS allele from unrelated plants. Several petunias homozygous forS ₁, and exhibiting various levels of PSC as determined by self seed set, progeny tests and temperature treatments, were used as pollen parents. The mean seed set of these crosses produced a ranking of the pollen parents which reflected the PSC levels obtained by other methods. The behavior of the F₁ and F₂ populations suggests that the pollen discriminating ability may be a simply inherited, dominant character in these plants. The styles of these unusual petunias illustrate the participation of the pollen tube in determining PSC.Scientific Journal Series Paper Number 10.479 of the Minnesota Agricultural Experiment Station     

Cytotaxonomical study ofSaxifraga seriesCeratophyllae s.l. (Saxifragaceae)

Chromosome numbers, mainly gametic, are given for 28 populations belonging to 13 species in ser.Ceratophyllae as circumscribed byWebb & Gornall. From a cytological point of view, we report or confirm the occurrence of aneuploidy in at least 8 of the 19 species recognized. This phenomenon, together with the technical difficulties inherent to the genus, is responsible for a number of incorrect or imprecise previous reports which are here questioned. Aneuploidy and the karyological instability behind it apparently reflect very active extant processes which might play an important role in the evolution of the group. Cytological and morphological data allow the recognition of four groups; one of them is likely to constitute a monophylectic group (ser.Ceratophyllae sensu stricto) but, the monophyly of the seriesCeratophyllae sensu lato is questioned.     

Pollen tube growth and the pollen‐tube pathway of Nymphaea odorata (Nymphaeaceae)

An important aspect of the evolution of carpel closure, or angiospermy, is the relationship between pollen tube growth patterns and internalization of the pollen‐tube pathway. True carpel closure, involving postgenital fusion of inner carpel margins, is inferred to have arisen once within the ancient order Nymphaeales, in the common ancestor of Nymphaeaceae. We studied pollen tube development, from pollination to fertilization, in a natural population of Nymphaea odorata, using hand pollinations and timed flower collections. Pollen germinates in stigmatic secretions within 15 min and pollen tubes enter subdermal transmitting tissue within an hour, following wide intercellular spaces towards the zone of postgenital fusion. At the zone of fusion they turn downwards to grow in narrow spaces between interlocked cells and then wander freely to ovules within ovarian secretions. The pollen‐tube pathway is 2–6 mm long and upper ovules are first penetrated 2.5 h after pollination. Pollen tubes grow at rates of approximately 1 mm/h whether in stigmatic fluid, transmitting tissues or ovarian secretions. Pollen‐tube pathways are structurally diverse across Nymphaeales, yet their pollen tubes have similar morphologies and rapid growth rates. This pattern suggests pollen tube growth innovations preceded and were essential for the evolution of complete carpel closure. © 2010 The Linnean Society of London, Botanical Journal of the Linnean Society, 2010, 162 , 581–593.     

Action of the Style Product of the Self-Incompatibility Gene of Nicotiana alata (S-RNase) on in Vitro-Grown Pollen Tubes

The products of the S-locus expressed in female tissues of Nicotiana alata are ribonucleases (S-RNases). The arrest of growth of incompatible pollen tubes in styles may result from entry of the S-RNase into the pollen tube and degradation of pollen tube RNA. We investigated the action of isolated S-RNases on pollen tubes grown in vitro and found that S-RNase is taken up by the pollen without substantial alteration. The S-RNases inhibit incorporation of exogenously added radioactive amino acids into protein by the germinated pollen. The S-RNases also inhibit in vitro translation of pollen tube RNA in a wheat germ cell-free extract. We found no evidence for a specific mRNA substrate for the S-RNases, which implies that if RNase activity is involved in the control of self-incompatibility, allelic specificity is more likely to depend on the selective uptake of S-RNases into pollen tubes or their selective activation or inactivation by pollen factors, rather than cleavage of a specific substrate. Heat treating S2-RNase largely destroys its RNase activity but increases its inhibitory effect on in vitro pollen tube growth. This effect is not due to an increased uptake of S2-RNase by the pollen but is associated with a greatly enhanced accumulation of S2-RNase on the outer surface of the pollen grains.     

On the gynodioecious polymorphism in Saxifraga granulata L. (Saxifragaceae)

Sexual and vegetative fitness components in hermaphrodite and female plants of the self-compatible, perennial herb Saxifraga granulata are compared using material derived from a gynodioecious population in northern England.
Females produced only 57% as many seeds as hermaphrodites, but their ovule offspring were 1.28 times as fit as those of hermaphrodites, and females were more vegetatively vigorous. The advantages to females in ovule offspring quality and in vegetative reproduction counteract their disadvantages in pollen and seed production and therefore probably play a role in the maintenance of the gynodioecious polymorphism. Pollination ecology, resource reallocation and inbreeding depression all appear to contribute to the observed sex differences in fitness.     

鹤顶兰花粉管在子房中的生长途径

运用扫描电镜对鹤顶兰(Phaiustankervilliae(Aiton)Bl.)花粉管在子房内的生长途径进行了观察。结果表明:花粉管在子房中的生长途径可以分为3个阶段:(1)沿子房壁轴向生长阶段,从授粉开始至大孢子母细胞四分体时期,花粉管经过合蕊柱到达子房,经由胎座基部沿子房壁轴向生长;(2)沿子房径向生长阶段,二核胚囊之后,花粉管在胚珠之间穿梭,以径向生长为主;(3)朝珠孔定向生长阶段,胚囊成熟时,花粉管朝珠孔定向生长进入胚囊。实验结果说明花粉管的定向生长受胚珠的分子信号调控。     

唐古特虎耳草谱系地理学研究

利用叶绿体基因(trnL-trnF和rpl16)对青藏高原地区的18个唐古特虎耳草(Saxifraga tangutica Engl.)居群(209个个体)进行谱系地理学研究,以揭示唐古特虎耳草的现有遗传结构及其历史演化过程。结果表明:(1)从209个个体中共检测到74个单倍型,且只有单倍型H5在居群中广泛分布,71.62%的单倍型为居群特有单倍型。(2)分子变异分析(AMOVA)显示,91.85%的遗传变异来源于居群内,居群间遗传分化不明显(F_(ST)=0.081);遗传分化系数N_(ST)(0.109)大于G_(ST)(0.097,P0.05)但不显著,表明唐古特虎耳草在其整个分布范围内没有明显的谱系地理结构。(3)中性检验表明,Tajima’s D(-2.045 07,P0.05)和FuLi’D*值(-3.629 27,P0.05)均为显著的负值,结合单峰的错配分布曲线,表明该物种经历过近期扩张。研究推测,唐古特虎耳草在第四纪冰期时可能存在多个微型避难所,由于第四纪冰期气候的反复波动,使得原来连续的居群片段化,避难所内的居群单独进化,从而形成了大量特有单倍型;唐古特虎耳草居群对第四纪冰期气候波动的反应可能更多的表现为垂直海拔高度的迁移,而非大规模的水平迁移。     

Pollen Tube Growth and Seed Set of Banksia coccinea R.Br. (Proteaceae)

Controlled hand pollinations and field observations were usedin conjunction with fluorescence and scanning electron microscopyand fruit and seed set to investigate the timing of stigma receptivity,pollen tube growth and self-incompatibility in relation to fertilityin B. coccinea. The species showed both protandry and partialself-incompatibility. Peak stigma receptivity as measured bypollen germination was recorded at 3 d after anthesis and maximumproduction of stigmatic exudate at 6 d. Pollen tubes reachedthe base of the style by 6 d after pollination. A 5 x 5 diallelexperiment was conducted and the results measured by pollentube growth. Self-pollinations generally resulted in poorertube growth than crosses and there was significant specificand general combining ability as well as reciprocal effects.Cross-pollination resulted in improved fruit set and seed toflower ratio over both selfing and open pollination. Spatiallimitations to fertility due to infructescence size were alsorecorded, but the combination of outcrossing mechanisms andspatial limitation did not entirely account for the low fertility.It is suggested that environmental conditions and the availabilityof resources may also exert an influence. Banksia coccinea R.Br., scarlet banksia, Proteaceae, pollination, stigma receptivity, pollen tube growth, self-incompatibility, breeding system, seed set     

Haplotype and ribotype diversity of Saxifraga cuneifolia s. l. (Saxifragaceae)

Saxifraga cuneifolia L. (sect. Gymnopera, Saxifragaceae) is a plant distributed in the main mountain ranges of southern Europe, from the eastern Pyrenees to the eastern Carpathian. Currently, two subspecies are recognized based on morphological characteristics: S. cuneifolia subsp. cuneifolia grows in the Maritime Alps and North Apennines and S. cuneifolia subsp. robusta is located in the remaining area of distribution. A more delicate form and a smaller number of flowers in S. cuneifolia subsp. cuneifolia are the morphological characteristics that differentiate this subspecies from the widespread S. cuneifolia subsp. robusta. To explore the genetic diversity and the subspecific geographic patterns of S. cuneifolia s. l. we conducted a molecular study of nuclear and plastid sequences. Samples of S. cuneifolia s. l. have been analysed throughout the distribution area of this species. Our results, based on nuclear (ITS) and plastid (rbcL, trnL–F, and psbA–trnH) markers, showed a genetic characterization of both subspecies presenting discriminant haplotypes and ribotypes that confirm the current subspecific systematics.