Geographic variations in seed dispersal by ants: are plant and seed traits decisive?

The effect of local ant species on the dispersal success of a myrmecochorous plant, Helleborus foetidus, was analyzed in two populations of the Iberian Peninsula (Caurel and Cazorla, respectively). The contribution of the various local ant species to dispersal was very unequal. While 5 and 19 ant taxa visited the plants of Caurel and Cazorla, respectively, most removal activity (67 and 80%) was performed by two species only (Formica lugubris and Camponotus cruentatus, respectively). Visits by dispersers were also unequally distributed between neighboring plants. While some plants were always visited during the period of seed release, others were never visited. A regression model indicated that this pattern might be explained by two plant traits: ants preferred to visit plants that released more seeds and whose elaiosomes were richer in oleic acid. Although it has long been known that this compound triggers removal by ants, it is the first demonstration that quantitative variations in elaiosome traits contribute to variation in dispersal success. Finally, other variables being equal, morphological traits (seed size, elaiosome size, and elaiosome/seed size ratio) did not affect ant behavior. Although myrmecochory has long been considered a diffuse interaction, our results support the idea that, at local scale, a limited number of ant species may be decisive to its evolution.     

A note on the effects of tropical weed decomposition on seed germination and seedling growth of some agricultural crops

Greenhouse and laboratory experiments were carried out to evaluate the allelopathic effects of Euphorbia heterophylla, Cyperus rotundus and Digitaria sanguinalis on seed germination and seedling growth of tomato, soko (feather cockscomb), pepper and sorghum. Significant reductions in germination and seedling growth of tomato, pepper and sorghum were observed when grown in pots containing increasing quantities of decomposed weeds. Decomposed Euphorbia heterophylla had the least suppressive effect on crop seeds, and soko seeds germinated without significant suppression. Seedling growth of sorghum was severely retarded when allowed to grow in leachates obtained from soil containing decomposed weeds.     

不同微塑料赋存环境对小麦萌发与幼苗生长影响研究

为揭示不同微塑料赋存环境对小麦（Triticum aestivum L.）种子萌发和幼苗生长的影响,选取聚丙烯（Polypropylene,PP）、高密度聚乙烯（HDPE）和聚乳酸生物可降解塑料（PLA）3种微塑料,并分别设置3种微塑料粒径（150μm、1000μm和4000μm）和质量浓度（0.1g/kg、0.5g/kg和1g/kg）在自然环境条件下开展盆栽试验,通过统计分析和方差分析对不同微塑料组合处理下小麦种子的发芽率、发芽抑制率、生长特征及幼苗生长趋势进行分析.结果表明：不同微塑料胁迫下均降低了小麦种子平均发芽水平,小麦种子平均发芽抑制率呈现出HDPE>PLA>PP;微塑料的粒径和质量浓度处于中粒径和中浓度条件下对小麦种子的平均发芽抑制率最大分别为5.65%和4.55%,而处于低或高粒径和质量浓度条件下微塑料对小麦种子发芽的抑制作用有所减弱;微塑料粒径对小麦种子发芽率的影响作用较大,微塑料类型和质量浓度次之.与对照组相比微塑料HDPE和PLA对小麦种子生长特征指标（发芽势、发芽指数、活力指数和平均发芽时间）的影响作用比微塑料PP的影响作用更加显著,其中小麦种子活力指数受微塑料类型、粒径和质量浓度的影响最为显著.对照组和不同微塑料类型对小麦幼苗平均生长速率呈现出CK>PP>HDPE>PLA,而不同微塑料对小麦幼苗生长的抑制作用呈现为PLA>HDPE>PP,微塑料类型对小麦幼苗生长的影响作用最显著.     

微塑料与铅复合污染对玉米种子萌发与生长的影响

为探明微塑料和重金属复合污染对作物种子萌发与生长的影响,以玉米种子为对象,探究了铅(Pb)和3种微塑料[聚乙烯(PE)、聚丙烯(PP)、聚氯乙烯(PVC)]不同单一及复合浓度暴露对玉米种子萌发及其生长的影响.结果表明,Pb单一暴露对玉米种子发芽的抑制总体上随其浓度的升高而增强;与CK相比,500、 1 000和1 500 mg·L^-1PE暴露显著抑制玉米种子发芽,100 mg·L^-1和300 mg·L^-1暴露影响不显著(第5 d除外);所有PP浓度暴露均显著抑制玉米种子发芽;相比PP和PE,PVC单一暴露对玉米发芽抑制较轻.Pb和3种微塑料复合暴露均显著抑制玉米种子发芽,随着时间的增长,抑制逐渐减弱.玉米种子的发芽指数、发芽势和活力指数均随着Pb及3种微塑料单一暴露浓度升高而减小,且在Pb和PE复合暴露下较CK显著减小,但在Pb和PP、 Pb和PVC复合暴露下均无显著变化;3种微塑料中,PVC对玉米种子活力影响相对最轻.10 mg·L^-1Pb和100 mg·L^-1的3种微塑料单...     

聚苯乙烯纳米塑料(PSNPs)对大豆(Glycine max)种子发芽和幼苗生长的影响

为探讨微塑料对农作物的生态毒理效应，研究了两种粒径（20 nm和100 nm）聚苯乙烯纳米塑料（Polystyrene Nanoplastics，PSNPs）在6个浓度梯度（0、50、100、200、500和1000 mg·L^-1）下对大豆（Glycine max）种子发芽和幼苗生长的影响.结果表明，两种粒径PSNPs均抑制种子发芽能力（发芽势、发芽指数、活力指数、平均发芽速度和发芽率），其中，发芽抑制率在一定程度上与暴露时间呈负相关关系.20 nm和100 nm PSNPs对根长、芽长和苗长的影响整体呈"低浓度促进，中高浓度抑制"的规律.20 nm PSNPs对根尖数量无显著影响，而100 nm PSNPs则表现出促进作用.20 nm和100 nm PSNPs对根直径和干重均有抑制作用.综上，PSNPs的植物毒性与粒径和浓度密切相关，在中等浓度（200 mg·L^-1）时，其对大豆幼苗生长的毒害作用最大.     

Oviposition strategies of conifer seed chalcids in relation to host phenology

Insects are considered the most important predators of seed cones, the female reproductive structures of conifers, prior to seed dispersal. Slightly more than 100 genera of insects are known to parasitize conifer seed cones. The most diverse (i.e., number of species) of these genera is Megastigmus (Hymenoptera: Torymidae), which comprises many important seed pests of native and exotic conifers. Seed chalcids, Megastigmus spp., lay eggs inside the developing ovules of host conifers and, until recently, oviposition was believed to occur only in fertilized ovules. Ovule development begins just after pollination, but stops if cells are not fertilized. The morphological stage of cone development at the time of oviposition by seed chalcids has been established for many species; however, knowledge of ovule development at that time has been documented for only one species, M. spermotrophus. Megastigmus spermotrophus oviposits in Douglas-fir ovules after pollination but before fertilization. Unlike the unfertilized ovules, those containing a M. spermotrophus larva continue to develop, whether fertilized or not, stressing the need to broaden our understanding of the insect–plant interactions for this entire genus. To achieve this task, we reviewed the scientific literature and assembled information pertaining to the timing of oviposition and to the pollination and fertilization periods of their respective host(s). More specifically, we were searching for circumstantial evidence that other species of Megastigmus associated with conifers could behave (i.e., oviposit before ovule fertilization) and impact on female gametophyte (i.e., prevent abortion) like M. spermotrophus. The evidence from our compilation suggests that seed chalcids infesting Pinaceae may also oviposit before ovule fertilization, just like M. spermotrophus, whereas those infesting Cupressaceae seemingly oviposit after ovule fertilization. Based on this evidence, we hypothesize that all species of Megastigmus associated with Pinaceae can oviposit in unfertilized ovules, whereas those exploiting Cupressaceae cannot, and thus oviposit only in already fully developed fertilized seeds. Furthermore, we predict that the presence of a larva in unfertilized ovules of all Pinaceae will influence the development of the female gametophyte by preventing its abortion. This influence on the Pinaceae can be interpreted as an ability to parasitize any of the potential seeds present in a seed cone, and as such represents a much more efficient oviposition strategy than searching and locating only fertilized seeds. Concomitantly, this ability has likely led to an overestimation of the impact of the species of seed chalcid infesting Pinaceae on seed production.     

Seed drops and caches by the harvester ant Messor barbarus: do they contribute to seed dispersal in Mediterranean grasslands?

 To determine whether the harvester ant Messor barbarus acts as a seed disperser in Mediterranean grasslands, the accuracy level of seed processing was assessed in the field by quantifying seed drops by loaded foragers. In the vicinity of exploited seed patches 3 times as many diaspores were found as in controls due to seed losses by foragers. Over trails, up to 30% of harvested seeds were dropped, singly, by workers but all were recovered by nestmates within 24 h. Seeds were also dropped within temporary caches with very few viable diaspores being left per cache when ants no longer used the trail. Globally, ant-dispersed diaspores accounted for only 0.1% of seeds harvested by M. barbarus. We discuss the possible significance for grassland vegetation of harvester-ant-mediated seed dispersal. Received: 26 January 2000 / Accepted in revised form: 20 June 2000     

Germination of Deyeuxia angustifolia as affected by soil type, burial depth, water depth and oxygen level

Deyeuxia angustifolia is a dominant species in the Sanjiang Plain, the largest freshwater marsh wetland in China. However, this species has declined because of great environment changes. In the effects of sea-level rise and greenhouse gas, magnitude and frequency of flood and rate of deposit in wetland increased greatly. Thus, in this paper, effects of soil environment (soil type, burial depth, water depth and oxygen concentration) on seed germination of D. angustifolia were investigated. Both soil type and burial depth affected seedling emergence. Seedlings emerged through 1-, 2-, 4-mm sand, and at 1-and 2-mm mud. Regardless of soil, seedling emergence time increased and emergence percentage decreased with increasing burial depth. For example, seedlings started to emerge on the 4th day and the final emergence percentage was 30% when covered with1-mm mud, whereas the initial emergence time was on the 6th day and the final emergence percentage was only 8% when covered with 2-mm mud. At the same burial depth, emergence percentage was higher in sand than in mud. Increasing water depth inhibited seed germination. Germination percentage in 2-cm water level (20%) was 1.2 times higher than that in 12-cm water level (9%). Oxygen concentration also affected germination. In the 21% oxygen concentration, germination was started on the 2nd day and the germination percentage was 36%, while seed started to germinate on the 4th day and the germination percentage was only 3% at the 7% oxygen concentration. These data indicate that seed germination of D. angustifolia was highly sensitive to soil type, burial depth, water depth, oxygen concentration, and these responses may be an explicit indication of maladjustment to environment changes, and an important reason accounting for the decline of this species in the Sanjiang Plain.     

滨海湿地盐沼植被修复中的种子产品制作过程及其有效性

针对当前滨海湿地种子流在潮汐过程中种子有效沉降降低、土壤种子库分布不均、定植效率低和栖息地裸斑现象等关键问题,以黄河三角洲盐沼湿地的盐地碱蓬（Suaeda salsa）为修复物种,研发一种应用于滨海湿地盐沼植被修复的固态种子产品,阐述固态种子产品的制作过程,并开展三种不同保水材料为核心材料（聚丙烯酰胺PAM,羟乙基纤维素HEC,羟丙基甲基纤维素HPMC）的种子产品的有效性研究。研究结果表明：（1）包装制作后的盐地碱蓬种子的吸水倍率和保水持续时间显著高于普通天然种子,表现出良好的吸水保水特性;（2）包装后的盐地碱蓬种子受盐度梯度变化的影响较小,表现出良好的耐盐抗性;（3）包装后的盐地碱蓬种子具有更高的萌发潜力,而不同保水材料的种子产品包装对盐地碱蓬种子萌发的促进作用表现为PAM>HPMC>HEC>天然种子;（4）包装后的盐地碱蓬种子具有更短的悬浮时间和较快的平均沉降速度,表明了种子产品增强了盐地碱蓬种子抵抗水动力干扰的潜力,其中以PAM为保水材料的种子产品可以直接、快速地沉降。本文首次将种子产品技术应用于滨海湿地盐沼植被修复中,为种子产品技术在退化滨海湿地的植被修复领域的应用与实践提供了理论基础。     

气温和土壤湿度变化对3种典型荒漠植物种子发芽的影响

植物种子发芽对气温和土壤湿度变化的响应是气候变化对荒漠生态系统影响的关键. 为了分析荒漠植物种子发芽对气温和土壤湿度变化的响应趋势,选择典型荒漠植物沙拐枣(Calligonum mongolica)、泡泡刺(Nitraria sphaerocarpa)和麻黄(Ephedra przewalskii),在人工气候箱中分析在3种土壤湿度(3%,5%和15%),2种气温(16 ℃/22 ℃ 和21 ℃/27 ℃)和浸种(12 h)与不浸种处理下种子的发芽趋势. 结果显示:沙拐枣和泡泡刺种子的发芽率随土壤湿度的增加而增加,浸种下随气温的升高其发芽率增幅比不浸种下大;不浸种下气温升高麻黄种子发芽率增幅比不浸种下大,浸种下土壤湿度为3%时气温升高其发芽率增幅比土壤湿度为5%和15%时大(P<0.05). 3种植物种子发芽指数随气温的升高和土壤湿度的增加而增加,不浸种下气温升高沙拐枣和泡泡刺种子的发芽指数显著增加,浸种下其发芽指数增幅不大;不浸种下气温升高麻黄种子发芽指数增幅较小,浸种下其发芽指数增幅较大. 3种植物种子均在培养初期发芽速率增加较快. 荒漠植物种子发芽受气温和土壤湿度的共同影响,不同植物种子发芽对这些因素的变化有不同的响应.      

Post-dispersal weed seed predation by avian and non-avian predators

Seed predation by avian and non-avian predators was quantified in the boundaries and cropped areas of cereal fields by presenting known quantities of seed with and without exclusion cages. Predator encounter-rates with the dishes exceeded 99%. Birds removed on average 6.7% seed from the dishes during the seven-day trials compared to 51% by non-avian predators. A comparison was made of the causal factors responsible for predation of Avena fatua, Chenopodium album and Cirsium arvense seeds. A. fatua seeds were preyed most heavily by both avian and non-avian predators. Seed removal by birds was greater in the cropped area than in the field boundary, non-avian predators being generally more active in the field boundary. Seed predation by birds was greater in spring than in any other season, whilst losses to other animals were greater during autumn and winter. Although, birds were not the main seed predators in cereal fields, they may contribute to weed seed depletion, of relevance to reduced-input farming systems where herbicides use is restricted.     

Mechanical defence in seeds to avoid predation by a granivorous ant

Harvester ants have traditionally been considered as seed predators that negatively affect plants. In some cases, however, they can also act as positive seed dispersers. During field observations, we noted that a portion of Psoralea bituminosa seeds that were collected and carried to the nest by the granivorous harvester ant Messor barbarus were discarded intact in refuse piles outside the nest. We analyzed and compared the physical characteristics of size, mass and toughness in P. bituminosa seeds from two different origins: intact seeds found in the ant's refuse piles and seeds collected directly from the plants. Seeds from refuse piles were similar in width but lighter and tougher than seeds from the plant. Our results point to a mechanical defence based on seed toughness to avoid predation by M. barbarus and suggest that an elevated proportion (~69%) of the seeds produced by P. bituminosa could be too tough to be consumed by this ant. These transported but uneaten seeds could benefit by being moved far from the mother plant and this could act as a selective evolutionary pressure towards tough seeds.     

酸雨对水稻、小麦和油菜种子萌发的影响

采用pH2.0、2.5、3.0、3.5、4.0与5.0模拟酸雨处理水稻、小麦和油菜种子的实验结果表明,3者在pH 2.0时不发芽,在pH 2.5时只有水稻和小麦异状发芽,pH≥3.0时,水稻、小麦和油菜种子的发芽率、发芽势、发芽指数、活力指数都与pH值显著正相关,水稻和小麦的异状发芽率则随pH值上升而降低.部分生理指标的实验结果表明,水稻、小麦和油菜的吸水值、呼吸速率和贮藏物质运转效率也与pH值显著正相关,水稻与小麦贮藏物质消耗率与pH值显著正相关,而油菜与pH值显著负相关;水稻、小麦和油菜的芽、根长抑制指数都与pH值显著负相关.酸雨胁迫下,种子各项指标变幅为水稻＜小麦＜油菜,这表明抗酸雨胁迫能力为水稻＞小麦＞油菜.     

1,4-二氯苯对小白菜种子萌发和幼苗的毒性

采用室内模拟染毒法,研究1,4-DCB(1,4-二氯苯)对3种小白菜种子(矮青6号、上海青和秦白2号)萌发和蔬菜幼苗的毒性作用. 结果表明:1,4-DCB作用后,3种小白菜种子的发芽势、种子发芽率、发芽指数和活力指数均降低,并呈现一定的剂量-效应关系;种子受1,4-DCB胁迫后,小白菜幼苗的根和苗生长都受到了抑制,根长和苗高降低,根苗比显著下降;随着ρ(1,4-DCB)的升高,受胁迫后的幼苗叶片的w(游离脯氨酸)呈现先升高后降低的趋势,而MDA(丙二醛)含量逐渐升高,与ρ(1,4-DCB)呈正相关;幼苗叶片SOD(超氧化物歧化酶)活性在ρ(1,4-DCB)为5 mg/L时升高,在ρ(1,4-DCB)为10和50 mg/L时明显降低;与对照组相比,各处理组幼苗叶片POD(过氧化物酶)活性均升高,但是ρ(1,4-DCB)为50mg/L时酶活性比ρ(1,4-DCB)为10 mg/L时下降. 表明水体中1,4-DCB污染对蔬菜种子萌发和幼苗生长产生毒性,其毒性机制与膜脂质过氧化有关.      

Response of peroxidase and catalase to acid rain stress during seed germination of rice, wheat, and rape

Seed germination of plants with various acid-resistance display different responses to acid rain. To understand the reason why such differences occur, the effects of simulated acid rain (pH 2.5–5.0) on the activities of peroxidase (POD) and catalase (CAT) during seed germination of rice (O. sativa), wheat (T. aestivum), and rape (B. chinensis var. oleifera) were investigated. Results indicated that the maximum change in activities of CAT and POD by acid rain treatment with different acidity and time in relation to the referent treatment without acid rain, was in the order: rice (28.8%, 31.7%) < wheat (34.7%, 48.3%) < rape (79.3%, 50.0%). The pH level for which the treatment with acid rain did not cause significant difference (p < 0.05) was in the order: rice (3.5).wheat (4.0).rape (5.0). Moreover, the change in activity of POD was higher than that of CAT, which showed that POD was more sensitive to acid rain stress than CAT. The difference in the ability of POD and CAT in removing free radicals was one reason why the germination indexes of these three species behaved differently. __________ Translated from Environmental Science, 2005, 26(6): 123–125 [译自: 环境科学]     

The action spectrum for maximal photosensitivity of germination

 Fruits of Garden Lettuce, imbibed in 0.01 M KNO₃, were depleted of maternal active phytochrome B by saturating deep-red exposure and photosensitized by chilling for 1 week at 4  °C. Twenty saturated fluence-response curves for photoinduced germination were elaborated between 300 and 800 nm, using exposure periods from 6 to 600 s at 22.5  °C; there is linear and closely parallel regression in the logarithmic probability net. The reciprocals of the half-response fluences obtained gave the apparent conversion spectrum of the controlling pigments and this was corrected for the transmittance of the seed-coat. It is a phytochrome spectrum of P _r with photoconversion cross-sections of 1.2·10⁹ and 4.5·10³ m² mol^–1 at 666 and 800 nm, respectively. This means that for half-saturated germination of sensitized seed, fewer than 1 out of 200,000 phytochrome A molecules have to be photoconverted to P _fr, and no photo-reversibility by deep-red was found. Therefore, all spectral colours of nightly moon- or skylight should stimulate the germination of sensitized weed seeds if they are exposed at the soil surface between sequential tillage operations. Received: 28 April 2000 / Accepted in revised form: 28 July 2000     

DAT-IAT工艺处理城市污水的生态风险评估

采用常规水质指标检测和种子发芽及根伸长抑制的陆生生态毒理方法,评价了DAT-IAT工艺处理过程中各工艺阶段污水的生态风险.常规水质指标检测结果显示,COD和SS可以达标排放,而氮的形态由进水时的NH⁺₄-N转变为DAT、出水中的NO^-₃-N,只是形态发生了变化,并没有从污水中去除,虽然达到了设计的NH⁺₄-N去除要求,可以达标排放,但是并未降低受纳水体发生富营养化的生态风险.生态毒理指标检测结果显示,进水、DAT、出水各阶段污水中COD对小麦种子发芽的半数抑制浓度逐渐降低,分别为249.3、 165.3和161.2 mg·L^-1;对小麦根伸长的半数抑制浓度也逐渐降低,分别为257.6、 154.0和142.1 mg·L^-1,表明DAT-IAT工艺处理过程中污水的生态毒性变化趋势为逐渐增大,处理后的出水存在一定的生态风险,有进一步进行生态风险调控的必要性.在相同水质条件下,小麦根伸长抑制率比小麦种子发芽抑制率更显著,可以把小麦根伸长作为评价污水处理生态风险的敏感指标之一.     

乙酰甲胺磷和汞单一及复合污染对黄瓜发芽及幼苗生长的影响

有机磷农药和重金属是目前我国土壤中较常见的污染物。本文以黄瓜种子为供试作物,研究了重金属汞与农药乙酰甲胺磷单一污染及复合污染对其发芽率以及幼苗生长的毒害效应。结果表明,Hg2+与乙酰甲胺磷单一污染及复合污染对黄瓜种子的发芽率、芽长和根伸长的生态毒性敏感程度依次为:根伸长>芽长>发芽率,Hg2+与乙酰甲胺磷单一污染及复合污染对黄瓜幼苗的毒害效应均较明显。复合污染对黄瓜种子发芽指标和幼苗生长的抑制效应表现为协同作用,其结果是复合污染的整体生态毒性效应增强。研究结果为进一步研究农药和重金属复合污染的生态毒性提供基础资料。     

Role of post-dispersal seed and seedling predation in establishment of dandelion (Taraxacum agg.) plants

Dandelion Taraxacum agg. (formerly Taraxacum officinale G.H. Weber ex Wiggers) is a common weed species associated with pastures, grasslands and no-tillage cropping systems throughout its native range in Europe, and more recently introduced into North America, Australasia and elsewhere. Following wind-dispersal from the parent plant, its seeds are subject to predation from a host of invertebrate predators. Similarly, seedling predation may also significantly limit dandelion recruitment. Although such post-dispersal mortality is central to our understanding of dandelion population dynamics and therefore weed control, the precise spatio-temporal role played by different putative seed and seedling predators is poorly understood. Here we studied how seed viability, and seed and seedling predation influenced dandelion recruitment at two contrasting sites in central Europe. The abundance in the field and seed and seedling consumption in the laboratory were determined for the main groups of predators—ground beetles (Coleoptera: Carabiade), terrestrial isopods (Isopoda: Oniscidea) and molluscs (Gastopoda: Pulmonata). At particular sites, seed viability and seedling predation were negatively correlated while the percentage of seeds that succumbed to seed predation was similar. Combined factors accounted for the death of 98% and 87% of exposed seeds. Ground beetles (particularly Amara spp.) and terrestrial isopods (Armadillidium vulgare) were efficient and dominant seed predators, while slugs (Arion lusitanicus) and isopods were important predators of seedlings. While there was no seasonal trend in the intensity of seed predation it decreased towards autumn in parallel with the feeding activity of the declining population of A. lusitanicus. The mortality factors thus varied in their importance, largely between sites and less with the course of the season. Although seed inviability, seed and seedling predation did not stop the recruitment of dandelion seedlings they are crucial factors limiting dandelion populations. Methods of increasing the efficiency of predation of seed as a means of managing weeds are worthy of further study, particularly in areas where dandelion is an invasive species.     

Factors affecting in vitro plant regeneration of the critically endangered Mediterranean knapweed (Centaurea tchihatcheffii Fisch et. Mey)

Habitat destruction has resulted in the extinction of many plant species from the earth, and many more face extinction. Likely, the annual endemic Mediterranean knapweed (Centaurea tchihatcheffii) growing in the Golbasi district of Ankara, Turkey is facing extinction and needs urgent conservation. Plant tissue culture, a potentially useful technique for ex situ multiplication, was used for the restoration of this ill-fated plant through seed germination, micropropagation from stem nodes, and adventitious shoot regeneration from immature zygotic embryos. The seeds were highly dormant and very difficult to germinate. No results were obtained from the micropropagation of stem nodes. However, immature zygotic embryos showed the highest adventitious shoot regeneration on Murashige and Skoog (MS) medium, containing 1 mg l⁻¹ kinetin and 0.25 mg l⁻¹ NAA. Regenerated shoots were best rooted on MS medium containing 1 mg l⁻¹ IBA and transferred to the greenhouse for flowering and seed set. As such, the present work is the first record of in vitro propagation of critically endangered C. tchihatcheffii, using immature zygotic embryos, and is a step forward towards conservation of this indigenous species.