Reproductive biology of Magnolia sinica (Magnoliaecea), a threatened species with extremely small populations in Yunnan,China

Magnolia sinica is one of the most threatened trees endemic to Southeast Yunnan. Based on our investigations, only 52 individuals and eight populations are found in the wild. M. sinica has been categorized as Critically Endangered on the IUCN Red List and identified as a “Plant Species with Extremely Small Populations (PSESP)”. Its fruit/seed set is very low and seedlings are rarely found in the wild. It is hypothesized that it may encounter obstacles to reproductive success. This study, therefore, focuses on its reproductive biology, knowledge of which is essential for effective conservation. Flowers of this species are protogynous and nocturnal, and possess a two-day rhythm of sexual presentation. For the first night of anthesis, the flowers are in the pistillate stage during which tepals open at dusk and close approx. 1 h later (except for the open outer ones). They remain closed until the next afternoon, when flowers, now in the staminate stage, re-open and remain so until the tepals drop. Nocturnal beetles enter into the flowers and remain trapped throughout the night as the flower closes, during which time they feed on tepals. Pollen-gathering bees are found to visit the re-opened flowers and the beetles are released during this stage. Two species of Pleocomidae and Curculionidae beetles appear to be effective pollinators.M. sinica is a self-compatible, pollinator-dependent species, and its fruit/seed set can be significantly increased by hand-pollination. No functional seed dispersers have been found in its extant natural habitats. These findings suggest that it may face both pollination and seed disperser insufficiencies in its current fragmented habitats, which may account for its low regeneration. Here we propose conservation strategies based on our findings.     

粉背含笑，云南东南部木兰科植物一新种

对产自云南东南部的木兰科植物一新种粉背含笑Magnolia glaucophylla Sima et H. Yu作了描述和绘图。该新种与棕毛含笑Magnolia fulva (H. T. Chang et B. L. Chen) Figlar接近,区别点在于其叶倒卵形、狭倒卵形或倒卵状椭圆形,侧脉每边19~24条;叶柄无毛;心皮20~26枚,成蓇葖时无毛,果瓣厚4~6 mm。     

极小种群广东含笑野外资源现状

为掌握极小种群广东含笑(Michelia guangdongensis)的种质资源现状并评估其濒危等级, 本文对广东含笑原生地——广东石门台国家级自然保护区及其邻近地区进行了系统的野外调查。结果表明: (1)目前发现广东含笑有4个野外分布区, 其中广东石门台国家级自然保护区红珠岩段和英德上天堂为首次报道。(2)船底顶是广东含笑最集中的分布区, 种群数量最多, 约有1,100株; 英德上天堂种群数量最少, 只有20株。(3)依据IUCN全球物种濒危等级评估标准, 广东含笑应当被评为濒危(EN, B1a + B2a)。     

Plant species with extremely small populations (PSESP) in China: A seed and spore biology perspective

Approximately one fifth of the world's plants are at risk of extinction. Of these, a significant number exist as populations of few individuals, with limited distribution ranges and under enormous pressure due to habitat destruction. In China, these most-at-risk species are described as ‘plant species with extremely small populations’ (PSESP). Implementing conservation action for such listed species is urgent. Storing seeds is one of the main means of ex situ conservation for flowering plants. Spore storage could provide a simple and economical method for fern ex situ conservation. Seed and spore germination in nature is a critical step in species regeneration and thus in situ conservation. But what is known about the seed and spore biology (storage and germination) of at-risk species? We have used China's PSESP (the first group listing) as a case study to understand the gaps in knowledge on propagule biology of threatened plant species. We found that whilst germination information is available for 28 species (23% of PSESP), storage characteristics are only known for 8% of PSESP (10 species). Moreover, we estimate that 60% of the listed species may require cryopreservation for long-term storage. We conclude that comparative biology studies are urgently needed on the world's most threatened taxa so that conservation action can progress beyond species listing.     

Effects of seed storage on germination of two succulent desert halophytes with little dormancy and transient seed bank

Seeds of both Salsola imbricata and Haloxylon salicornicum have high germination level and germination speed, and form a transient seed bank in nature. The impacts of storage period and condition on germination level and speed were assessed in the two species. Storage for three months significantly increased both germination level and speed of seeds stored under the different conditions, compared to that of fresh seeds. In both species, nine months storage did not affect germination percentage in cold storage seeds, but completely inhibited it in field seeds. Storage for longer time in room and warm temperatures resulted in significant reduction or complete inhibition in the germination of the two species, so this was more pronounced in H. salicornicum. Storage significantly increased germinate rate index of seeds stored in all conditions till 17 months in S. imbricata and till 12 months in H. salicornicum. In both species, fridge storage had little effects on final germination and germination speed of seeds incubated at the different temperatures, compared to fresh seeds. However, room temperature and warm storages significantly reduced final germination and germination speed at the different temperatures, so the reduction was more pronounced at 35 °C, especially in H. salicornicum.     

Seed mass and dormancy of annual plant populations and communities decreases with aridity and rainfall predictability

Several theoretical and empirical studies have examined the influence of environmental conditions on seed traits and germination strategies of annual species. A positive relationship between seed mass and dormancy has been described for annuals occupying climatically unpredictable ecosystems. Larger-seeded species tend to have higher seedling survival rates, while dormancy allows a bet-hedging strategy in unpredictable environments. Until now, these ideas have been addressed primarily for only one or a few focal species, without considering differences among populations and communities. The novelty of the present study lies in the population and community-level approach, where a comprehensive seed trait database including 158 annual species occurring along a gradient of rainfall variability and aridity in Israel was used to ask the following question: Does average seed mass and dormancy of annual populations and communities decrease with increasing aridity and rainfall unpredictability?Soil seed bank samples were collected at the end of the summer drought, before the onset of the rains, from four plant communities. Germination was tested under irrigated conditions during three consecutive germination seasons to determine the overall seed germinability in each soil sample. Seed mass was obtained from newly produced seeds collected at the study sites in late spring. The community level results showed that, in contrast to common theoretical knowledge, seed mass and dormancy of the dominant annual species decreased with increasing aridity and rainfall variability. Accordingly, a negative correlation was found between seed mass and seed germination fractions. The present study demonstrates that an analysis of seed traits along climatic gradients is significantly improved by approaches that target both population and community levels simultaneously. A critical evaluation sheds new light upon the selective pressures that act on seed ecology of annuals along a climatic gradient and facilitates formulation of more mechanistic hypotheses about factors governing critical seed traits.     

Seed viability of five wild Saudi Arabian species by germination and X-ray tests

Our objective was to evaluate the usefulness of the germination vs. the X-ray test in determining the initial viability of seeds of five wild species (Moringa peregrina, Abrus precatorius, Arthrocnemum macrostachyum, Acacia ehrenbergiana and Acacia tortilis) from Saudi Arabia. Usually several days were required to determine the viability of all five species via germination tests. However, X-ray test will give immediate results on filled/viable seeds. Seeds of all species, except Acacia ehrenbergiana and Acacia tortilis showed high viability in both germination (96–72% at 25/15 °C, 94–70% at 35/25 °C) and X-ray (100–80%) test. Furthermore, there was a general agreement between the germination (19%, 14% at 25/15 °C and 17% and 12% at 35/25 °C) and X-ray (8%, 4%) tests in which seed viability of Acacia ehrenbergiana and Acacia tortilis was very low due to insect damaged embryo as shown in X-ray analysis. Seeds of Abruspreca torius have physical dormancy, which was broken by scarification in concentrated sulfuric acid (10 min), and they exhibited high viability in both the germination (83% at 25/15 °C and 81% at 35/25 °C) and X-ray (96%) tests. Most of the nongerminated seeds of the five species except those of Acacia ehrenbergiana and Acacia tortilis, were alive as judged by the tetrazolium test (TZ). Thus, for the five species examined, the X-ray test was proved to be a good and rapid predictor of seed viability.     

Genetic diversity and structure of Rhododendron meddianum,a plant species with extremely small populations

Rhododendron meddianum is a critically endangered species with important ornamental value and is also a plant species with extremely small populations. In this study, we used double digest restriction-site-associated DNA sequencing (ddRAD) technology to assess the genetic diversity, genetic structure and demographic history of the three extant populations of R. meddianum. Analysis of SNPs indicated that R. meddianum populations have a high genetic diversity (π = 0.0772 ± 0.0024, H_E = 0.0742 ± 0.002). Both F_ST values (0.1582–0.2388) and AMOVA showed a moderate genetic differentiation among the R. meddianum populations. Meanwhile, STRUCTURE, PCoA and NJ trees indicated that the R. meddianum samples were clustered into three distinct genetic groups. Using the stairway plot, we found that R. meddianum underwent a population bottleneck about 70,000 years ago. Furthermore, demographic models of R. meddianum and its relative, Rhododendron cyanocarpum, revealed that these species diverged about 3.05 (2.21–5.03) million years ago. This divergence may have been caused by environmental changes that occurred after the late Pliocene, e.g., the Asian winter monsoon intensified, leading to a drier climate. Based on these findings, we recommend that R. meddianum be conserved through in situ, ex situ approaches and that its seeds be collected for germplasm.     

Seed dormancy and germination of five selected NATURA-2000 plant species from Croatia showing different germination strategies

Abstract

NATURA-2000, the ecological network of protected areas in the European Union that has been included in the Croatian legislation, defines Community important plant species with imperative on their conservation ex/in situ. In the Botanical Garden of the Faculty of Science, University of Zagreb, five NATURA-species have been selected as research subjects for germination study to shed light on the topic of their seed ecology and consequently advance their conservation efforts: Degenia velebitica, Scilla litardierei, Klasea lycopifolia, Ligularia sibirica and Genista holopetala. The freshly matured seeds of each species were exposed to cold or warm stratification in duration of four to sixteen weeks, and their germination was investigated through different regimes of incubation parameters, i.e. illumination (light/dark) and temperature (5, 15/6, 23?°C). All species had higher germination values after cold stratification, with the exception of G. holopetala. We concluded that D. velebitica, K. lycopifolia and presumably L. sibirica seeds has non-deep physiological dormancy while S. litardierei has deep complex morphophysiological dormancy and G. holopetala has physical dormancy. The observed patterns in seeds’ behaviour are consistent with the conditions in their natural habitats in Croatia and the knowledge of these patterns is vital for successful conservation strategies in the future.     

Seed flotation and germination of salt marsh plants: The effects of stratification, salinity, and/or inundation regime

We examined the effects of cold stratification and salinity on seed flotation of eight salt marsh species. Four of the eight species were tested for germination success under different stratification, salinity, and flooding conditions. Species were separated into two groups, four species received wet stratification and four dry stratification and fresh seeds of all species were tested for flotation and germination. Fresh seeds of seven out of eight species had flotation times independent of salinity, six of which had average flotation times of at least 50 d. Seeds of Spartina alterniflora and Spartina patens had the shortest flotation times, averaging 24 and 26 d, respectively. Following wet stratification, the flotation time of S. alterniflora seeds in higher salinity water (15 and 36 ppt) was reduced by over 75% and germination declined by more than 90%. Wet stratification reduced the flotation time of Distichlis spicata seeds in fresh water but increased seed germination from 2 to 16% in a fluctuating inundation regime. Fresh seeds of Iva frutescens and S. alternflora were capable of germination and therefore are non-dormant during dispersal. Fresh seeds of I. frutescens had similar germination to dry stratified seeds ranging 25-30%. Salinity reduced seed germination for all species except for S. alterniflora. A fluctuating inundation regime was important for seed germination of the low marsh species and for germination following cold stratification. The conditions that resulted in seeds sinking faster were similar to the conditions that resulted in higher germination for two of four species.     

Seed germination of high mountain Mediterranean species: altitudinal,interpopulation and interannual variability

The germination response of 20 species from high altitude Mediterranean climates, most of them rare endemics, was studied. Our main goal was to model the germination response of a complete set of Iberian high mountain species. The effect of temperature and other parameters, such as spatial and temporal short gradients, on germination were also evaluated. Some seed features (mass and size) were also related to the germination response. Finally, we tested the effect of cold-wet stratification pretreatment when germination was low under natural conditions. Seeds were collected at four locations from 1,900 to 2,400 m a.s.l. in the Sierra de Guadarrama (Spanish Central Range) over two consecutive growing seasons (2001–2002) and submitted to different temperatures and a constant photoperiod of 16 h light/8 h darkness. Most plants readily germinate without treatment, reaching an optimum at relatively high temperatures in contrast to lowland Mediterranean species. Seeds seem to be physiologically prepared for rapid germination even though these plants usually face very intense summer droughts after ripening and dispersal. Germination was also highly variable among altitudes, populations and years, but results were inconsistent among species. Such flexibility could be interpreted as an efficient survival strategy for species growing under unpredictable environments, such as the Mediterranean climate. Finally cold-wet stratification increased germination capacity in five of nine dormant species, as widely reported for many arctic, boreal and alpine species. In conclusion, high mountain Mediterranean species do not differ from alpine species except that a relatively high number of species are ready to germinate without any treatment.     

Seed dormancy release and germination characteristics of <i>Corispermum lehmannianum</i> Bunge,an endemic species in the Gurbantunggut desert of China

Seed dormancy release and germination of Corispermum lehmannianum Bunge were tested using various treatments: temperature, cold stratification, gibberelins (GA3), dry storage and sand burial. Results showed that temperature and light did not affect the germination of fresh seeds, cold stratification and GA3 could improve seed germination, whereas dry storage and sand burial did not. The germination percentage was highest at 35/20 °C after the cold stratification and GA3 treatments. Corispermum lehmannianum seeds were classified as non-deep, Type-2, physiological dormancy (PD), whose seed dormancy could be released by cold stratification and GA3.     

Physiology, morphology and phenology of seed dormancy break and germination in the endemic Iberian species Narcissus hispanicus (Amaryllidaceae)

Background and Aims

Only very few studies have been carried out on seed dormancy/germination in the large monocot genus Narcissus. A primary aim of this study was to determine the kind of seed dormancy in Narcissus hispanicus and relate the dormancy breaking and germination requirements to the field situation.

Methods

Embryo growth, radicle emergence and shoot growth were studied by subjecting seeds with and without an emerged radicle to different periods of warm, cold or warm plus cold in natural temperatures outdoors and under controlled laboratory conditions.

Key Results

Mean embryo length in fresh seeds was approx. 1·31 mm, and embryos had to grow to 2·21 mm before radicle emergence. Embryos grew to full size and seeds germinated (radicles emerged) when they were warm stratified for 90 d and then incubated at cool temperatures for 30 d. However, the embryos grew only a little and no seeds germinated when they were incubated at 9/5, 10 or 15/4 °C for 30 d following a moist cold pre-treatment at 5, 9/5 or 10 °C. In the natural habitat of N. hispanicus, seeds are dispersed in late May, the embryo elongates in autumn and radicles emerge (seeds germinate) in early November; however, if the seeds are exposed to low temperatures before embryo growth is completed, they re-enter dormancy (secondary dormancy). The shoot does not emerge until March, after germinated seeds are cold stratified in winter.

Conclusion

Seeds of N. hispanicus have deep simple epicotyl morphophysiological dormancy (MPD), with the dormancy formula C_1bB(root) – C₃(epicotyl). This is the first study on seeds with simple MPD to show that embryos in advanced stages of growth can re-enter dormancy (secondary dormancy).     

Seed germination of Lasia spinosa as a function of temperature,light, desiccation,and storage

Lasia spinosa seeds were not dormant at maturity in early spring. The most favorable temperatures for germination were between 25 and 30 °C, and final percentage and rate of germination decreased with an increase or decrease in temperature. When L. spinosa seeds were transferred to 25 °C, after 60 days at 10 °C (where none of the seeds germinated), final germination increased from 0% to 78%. Seeds germinated to high percentage both in light and in dark, although dark germination took more than twice as long as in the light. During desiccation of seeds at 15 °C and 45% relatively humidity, moisture loss decreased exponentially from 2.02 to 0.13 g H₂O g⁻¹ dry wt within 16 days, and only a few seeds (12%) survived 0.13 g H₂O g⁻¹ dry wt moisture content. Seeds stored at 0.58 g H₂O g⁻¹ dry wt moisture content at four constant temperatures (4, 10, 15, and −18 °C) for up to 6 months exhibited a well-defined trend of decreasing viability with decreasing temperature. Thus, we concluded that freshly harvested L. spinosa seeds are non-dormant and recalcitrant. Also, the seeds with 0.58 g H₂O g⁻¹ dry wt moisture content could be effectively stored for a few months between 10 and 15 °C although the most appropriate temperature for wet storage appears to be 10 °C, as it is close to the minimum temperature for germination and so there will be less pre-sprouting compared to 15 °C.     

Pollination and seed dispersal of Aquilaria sinensis (Lour.) Gilg (Thymelaeaceae): An economic plant species with extremely small populations in China

Pollination and seed dispersal in angiosperms have long been investigated in order to understand the coevolution of plants and animals. However, the signals from flowers and/or seeds to attract pollinators and/or seed dispersers have received comparatively little attention. In this study, the pollination biology and seed dispersal of the vulnerable agarwood plant Aquilaria sinensis (Lour.) Gilg, a traditional medicinal plant in China, was studied in its natural distribution range. The reproductive tactics of A. sinensis were studied in detail by employing various tests dealing with fruit set and also seed dispersal. Dynamic headspace extraction followed by GC-MS analysis was also performed in order to reveal the composition of floral scent. The results showed that noctuids and pyralids are the most effective pollinators of pollinator-dependent A. sinensis. The main compounds of the floral scent were (E, E)-α-Farnesene (61.9±3.2%), trans-Ocimene (16.6±1.2%), and Benzyl salicylate (4.6±1.1%). The results obtained from seed dispersal experiments indicate that hornets are effective seed dispersers and they may play an important role in long-distance seed dispersal of A. sinensis. Based on our findings, we recommend several protection methods for this threatened agarwood plant in China.     

极小种群野生植物抢救性保护面临的机遇与挑战

<正>过去50年,中国在生物多样性保护方面取得了一系列成就。但是,国家层面立法保护的植物仅有1999年国务院批准的《国家重点保护野生植物名录(第一批)》,之后并没有根据全国普查与各地调查报道的结果进行相应的更新。目前,最突出和最具有争议性的一个问题是,哪些物种才是最需要优先保护的?一些研究机构基于物种保护生物学的研究,认为应参考国际自然保护联盟(IUCN)对受威胁物种的等级划分标准来确定,但这些标准是基于全球尺度,并且大部分标准是依赖于对未来若干年的物     

木虱啮小蜂对枸杞、枸杞木虱的行为反应

木虱啮小蜂Tetrastichus sp.是枸杞木虱Paratrioza sinica Yang & Li若虫外寄生蜂,是控制枸杞木虱种群数量动态的重要因子之一。为查明其寻找寄主的信息素,应用四臂嗅觉仪、培养皿两种方法测定了枸杞Lycium barbarum L.的健康叶、虫伤叶、枸杞木虱若虫、若虫与叶复合体、卵与叶复合体和若虫分泌物等不同物质对木虱啮小蜂行为反应的影响。结果证明枸杞木虱若虫分泌物对木虱啮小蜂有明显的吸引作用,四臂嗅觉仪测定时有73%的雌蜂趋向分泌物材料区（P=0.0000）。枸杞健康叶, 虫伤叶及其水、正丁烷提取物, 卵与枸杞叶复合体对木虱啮小蜂无明显吸引作用。木虱若虫水提物及正丁烷提取物在培养皿中可吸引木虱啮小蜂并可引起刺探行为,但嗅觉仪测定时单独若虫并不吸引木虱啮小蜂,只有活体若虫与枸杞叶片复合体才对木虱啮小蜂有吸引作用（P=0.0004）。木虱啮小蜂通过与寄主若虫的接触,可提高其搜索效率。接触过寄主若虫的雌蜂第一次找到寄主的时间显著少于未曾接触寄主若虫的雌蜂,前者只用后者1/4的时间,说明该寄生蜂能对其寄主若虫进行学习。     

极小种群野生植物崖柏的生殖物候、传粉及胚胎发育研究

以濒危植物崖柏（Thuja sutchuenensis Franch.）为对象,对其生殖物候、传粉机制进行观察,并采用石蜡切片法对其胚胎发育过程进行研究。结果显示：崖柏于8月分化出大、小孢子叶球,次年3月传粉,为花粉无气囊、具传粉滴、胚珠直立型传粉机制,球果于10月开裂;显微观察发现,传粉期花粉进入珠孔后,贮藏在珠心上方的贮粉室内,同时珠心组织中分化出孢原细胞,进入雌配子体发育阶段,5月中旬,花粉管开始萌发,6月初完成受精,进入胚胎发育阶段,10月初,胚胎发育成熟。研究表明崖柏从大、小孢子叶球形成至种子成熟的整个发育过程中均存在败育,而胚珠败育及雌配子体游离核时期至幼胚发育期间的败育是其生殖障碍的主要原因。本研究获得了崖柏生殖生物学的基础资料,为其人工繁育和制定保护策略提供了重要依据。     

Seed germination and reproductive features of Lysimachia minoricensis (Primulaceae), a wild-extinct plant

Lysimachia minoricensis is one of the few Mediterranean endemic plants (Minorca, Balearic islands) that has gone extinct in the wild but which persists as extant germplasm or cultivated plants in several botanical gardens. Reproductive features (seed set, number of seeds per capsule, seed weight) and germination responses to constant temperatures, sea water and dry-heat pre-treatments were investigated to determine the extent to which they may have influenced the extinction of the species. Seed set in Lysimachia is not dependent on pollinators, suggesting a functional selfer breeding system. Most plants produced a large mean number of fruits (23.2) and seeds (466), and the mean production of seeds per individual was estimated to be almost 1,100. Overall, no highly specific requirements were observed for seed germination. Seed germination was not inhibited in the dark, and a high germinability (over 87% in all cases) was recorded in most experiments, with the exception of those performed at low temperatures (5 and 10 degrees C). These data suggest that fertility and seed viability were not the major causes of extinction. The high reproductive performance of L. minoricensis is in striking contrast to its status as a wild-extinct plant, suggesting that extrinsic factors were responsible for its extinction.     

Population differentiation and gene flow within a metapopulation of a threatened tree, Magnolia stellata (Magnoliaceae)

We examined genetic differentiation among eight local populations of a metapopulation of Magnolia stellata using 10 nuclear and three chloroplast microsatellite (nSSR and cpSSR) markers and evaluated the influence of historical gene flow on population differentiation. The coefficient of genetic differentiation among populations for nSSR (F(ST) = 0.053) was less than half that for cpSSR (0.137). An isolation-by-distance pattern was detected for nSSRs, but not cpSSRs. These results suggest that pollen flow, as well as seed dispersal, has significantly reduced genetic differentiation among populations. We also examined patterns of contemporary pollen flow by paternity analysis of seeds from nine seed parents in one of the populations using the nSSR markers and found it to be greatly restricted by the distance between parents. Although most pollen flow occurred within the population, pollen flow from outside the population accounted for 2.5% of the total. When historical and contemporary pollen flows among populations were compared, the levels of pollen flow seem to have declined recently. We conclude that to conserve M. stellata, it is important to preserve the whole population by maintaining its metapopulation structure and the gene flow among its populations.