In vitro culture of immature embryos from Koelreuteria bipinnata var. integrifoliola

For the mass production of Koelreuteria bipinnata var. integrifoliola with selected, hybrid or genetically engineered genotypes, one potentially desirable propagation strategy is based on embryo culture. The immature embryo development in vitro from K. bipinnata var. integrifoliola was studied under different conditions of embryo age, basic culture media and plant growth regulators. The results show that: 1) germination rate of grade 3 embryos in immature seeds with 0.6–0.8 cm diameter was 98.9%. The germination rate of grade 2 embryos in immature seeds with 0.4–0.6 cm diameter was 77.8% and the germination rate of grade 1 embryos in immature seeds with 0.4 cm diameter was 15.6%. 2) The amounts of macroelements in MS medium had no clear effect on the germination rate of immature grade 3 embryos and had a modest effect on plantlet growth, where the best medium was MS or 1/2 MS. The rates were all greater than 90%. 3) The germination rate of grade 3 embryos was greater than 87% when the medium contained a low concentration of NAA or no plant growth regulators at all and decreased markedly when BAP alone or BAP and NAA together were added to the media. We suggest that in vitro culture of immature embryos from K. bipinnata var. integrifoliola can be enhanced when a small amount of plant growth regulators is added. The addition of BAP has an adverse reaction to the germination and development of immature embryos.     

Improving the collection and germination of West African <Emphasis Type="Italic">Garcinia kola</Emphasis> Heckel seeds

Garcinia kola is highly used as chew-stick for dental care in West Africa, but it is nearly commercially extinct in nature. To help find methods to increase rate and percentage of germination, we investigated its seed germination responses by using several collections and pre-treatments. Seeds processed from 6-week old green fruits did not germinate, whilst those of 10-week old partly-mature, at the intermediate stage through full development, germinated 46%. Seeds from orange mature fruits reached maximum 54–62% germination. Radicles emerged 6 weeks after sowing and normal germination with emergence of the primary single shoot growth occurred after 12 weeks. There was no significant difference (P ≥ 0.05) between germination responses of seeds from Ghana and Cote d’Ivoire. Seeds did not germinate when dried to below ca. 30% MC. De-coating seeds or soaking intact seeds in a 70% ethanol solution increased germination to 82–92%, and also increased germination rate; however germination was not uniform. Because difficulties in germinating these seeds stem from the physical presence of the seed coats and the slow growth of the embryos contained therein, we recommend that fresh seeds from mature fruits be de-coated and immediately sown in preferably, an aerated medium (e.g., sawdust) for cultivation.     

Consequences of dry-season seed dispersal on seedling establishment of dry forest trees: Should we store seeds until the rains?

We examined the following hypotheses: (i) seeds of dry forest trees have high pre- and post-germination mortality by desiccation due to the time between seed dispersal and germination and to irregular rains at the onset of the rainy season; (ii) seedlings from seeds dispersed in the dry season which survive the dry spells are larger at the end of the first rainy season than those dispersed in the rainy season because the former have more time to grow. We evaluated the possible trade-off between few large seedlings (resulting from natural dispersal) × many small seedlings (resulting from delayed dispersal) on seedling survival during the dry season. We sowed eight tree species in a greenhouse in September, simulating the natural dispersal timing (before the rains), and in November, when rains are more constant. Because shading can counteract the effects of desiccation, we applied three levels of shade (10%, 40% and 72% of PPFD). From September 2005 to December 2006, we provided the daily precipitation of a median year from a major patch of dry forest in Central Brazil. At the end of the rainy season, a subset of seedlings was collected for growth measurements (dry mass) and the remainder was left to follow the dry season survivorship. The lower germination expected for seeds dispersed in the dry season and in full sun was not confirmed for species that had some dormancy. The delayed dispersal was advantageous for the initial establishment of fast germinating species, but it was irrelevant or even disadvantageous for others. Also, the greenhouse weather was certainly milder than the natural environment, reducing the potential for mortality by desiccation. The growth of the four species of higher dormancy were not affected by timing of seed dispersal, while three out of four fast germinating species had higher root biomass when dispersed in the dry season. The growth during the rainy season did not affect seedling survival during the dry season. Keeping seeds to sow when rain is constant might be a good strategy to increase the establishment of fast-germinating tree species.     

喜树种子中喜树碱和10-羟基喜树碱高效液相色谱法分析

建立了一种简便、快速、准确的测定喜树(Camptotheca acuminata Decne.)种子中喜树碱和10-羟基喜树碱含量的高效液相色谱分析方法;色谱条件为:采用日本KYA HIQ sil C18柱(250 mm×4.6 mm,5 μm),流速为1 mL/min,梯度洗脱程序为:在前 15 min流动相乙腈 -水的体积比由10 %线性增加至40%,在随后的3 min乙腈 -水的体积比线性降至10 %并保持恒定3 min,在21 min时停止该程序.检测器为二极管阵列检测器,喜树碱定量分析波长254 nm,10-羟基喜树碱定量分析波长266 nm,进样量10 μL.样品制备以60 %的乙醇作溶剂,在60 ℃下超声波提取喜树种子50 min.利用以上方法分别测定了喜树种子胚乳、胚轴、子叶和种皮中的喜树碱和10-羟基喜树碱含量,喜树碱的含量是胚乳＞胚轴,子叶＞种皮,10-羟基喜树碱的含量是胚乳＞种皮＞胚轴＞子叶.     

日本核用银杏枝叶生长及黄酮变化规律的研究

为 给银杏引 种和产 业化生产 提 供科 学依 据， 采用 日 本的 藤九 郎、 金兵 卫、 黄金 丸 和岭南４ 个大果 银杏的 标准 枝 和标 准 叶， 对 其 枝 叶生 产 及 黄硐 变 化规 律 进 行了 研 究。 结 果证 明，新梢长、 叶数 枝、叶柄 长、叶长 、叶 宽、 单叶 面 积、 单叶 干 重和 鲜 重 的年 变 化呈 Ｓ 型曲 线；新梢长 和叶数 枝分别 在７ 月４ 日和６ 月１４ 日达最 大值， 而 干重在７ 月底 达最 大值。 芽内 黄酮含量＞ 刚刚展开 的嫩叶 ＞ 成熟叶 ＞ 黄叶 ； ６ 月 初叶 内黄 酮含 量 下降 到原 初 的４０９ ％ ， 到１１ 月１６ 日又回升 到３０４ ％ ； 叶内黄酮 含量与 叶位及 叶龄 有关； 新梢 上部 叶及 幼 龄叶 单叶 面 积、干重、鲜 重低于下 部叶及老 龄叶， 但 黄酮含 量恰好相 反。     

Germination and storage characteristics of Prunus africana seeds

Prunus africana, from the moist highlands of sub-Saharan Africa, is subject to great exploitation because its bark is used in the treatment of prostate-related diseases. A better understanding of the biology of its seed could help the ex situ and in situ conservation of the species. Fifty per cent of fresh seeds (46.8% MC) from mature purple fruits harvested in Kenya germinated after 10 days incubation at 25°C. The germination rate was linearly related to constant temperature, giving a prediction for the minimum temperature for germination at ca. 0.6°C. All seeds germinated at 5°C during 4 months fully hydrated storage. P. africana seed is, thus, neither dormant nor chilling sensitive. The relationship between equilibrium moisture content and relative humidity was used to construct a sorption isotherm, against which seed desiccation and storage responses were assessed. Seventy six per cent of mature seeds survived drying to 4.5% MC (22% RH) compared to 17% of seeds from immature purple-green fruits. Although, 25–45% of mature seeds survived at 8–12% MC after 2 months, only 10% remained viable after 6-month storage at 5°C. This study proposes optimum conditions for the artificial regeneration of P. africana seeds and highlights the need for further experiments storing dry seeds at sub-zero temperatures.     

Seed dispersal phenology and dispersal syndromes in a subtropical broad-leaved forest of China

This study describes the dispersal phenology and syndromes in Gutianshan 24 ha plot in a subtropical broad-leaved forest of China. The 130 0.5 m² seed traps collected 69,115 mature seeds, representing 27 species (belonging to 24 genera, and 15 families) in 12 months. One marked peak in the number of seeds and species during the year was found in dry season (November). Zoochory was the most common dispersal syndrome (70.4%), followed by anemochory (18.5%), ballistic dispersal (11.1%). Among fruit types, berry (33%), capsule (22%), nut (18%), and drupe (11%) were common in the subtropical evergreen forest. In fruit color, brown was the commonest (40%), followed by dark brown (30%), black (15%), red (11%), and yellow (4%). Overall, the community level seed rain study revealed that one marked peak in seed number occurred in the middle of dry season; zoochory was the principal dispersal mode of woody plants in subtropical forest, and dry seasons favor seed dispersal by animal and wind.     

缙云山常绿阔叶林种子雨组成及其时空动态

【目的】分析缙云山常绿阔叶林种子雨组成及其时空动态,为常绿阔叶林种子生态学大尺度和长时序的研究提供数据支持,并为受损常绿阔叶林近自然恢复过程中的种源收集和种子撒播提供参考信息。【方法】 2014-01—2016-12,收集缙云山常绿阔叶林1 hm 2标准地中172个种子收集框内的种子,鉴定物种并计数,风干后称质量。根据果实类型及种子特征并结合野外实地观察,确定每个物种种子(果实)的散布方式。【结果】 3年间共收集到成熟种子36 298粒,种子雨密度为140.69粒·m -2 a -1;种子分属于19科21属27种,占群落木本植物物种总数的36.49%;种子雨具有明显的季节动态,在秋季出现高峰,其中有17个物种在干季(1—4和10—12月)成熟并散布,6个物种在湿季(5—9月)成熟并散布,4个物种在干、湿季均有散布;种子雨密度年际变异系数为0.36~1.73,空间变异系数为1.54~15.20。【结论】缙云山种子雨产量在常绿阔叶林中处于较低水平;优势种对群落种子雨产量贡献较大,种子雨物种组成与群落物种组成相似性较低;群落水平种子雨具有明显的季节动态,且存在很大的年际变异和空间变异,反映出亚热带常绿阔叶林种子雨时空动态的典型特征。在区域植被近自然恢复中的种子采集和撒播环节,应当充分参照不同物种的种子雨掉落的时空动态。     

The soil seed bank of the threatened plant <Emphasis Type="Italic">Magnolia stellata</Emphasis> is subordinate to the emergence of current-year seedlings

We studied seed bank formation of the threatened star magnolia, Magnolia stellata, to examine the early stage of regeneration. Forty-five seedling plots (2 × 2 m), each including a soil-sampling quadrat (40 × 40 cm), were established randomly under or around the crowns of mature M. stellata trees. Seeds of M. stellata were collected from each quadrat to a depth of 5 cm. Only four seeds of M. stellata were found (0.56 seeds/m²) and all were located under mature crowns. Current-year seedlings were abundant in water channels, on moss, or under mature crowns, suggesting that the seeds may require wet soil conditions for germination. Magnolia stellata seeds show considerable germination below the crowns of mature trees in the year following masting, while some seeds remain dormant in the soil. Considering the soil seed bank and the current-year seedling bank of M. stellata, a frequent supply of seed is essential for the regeneration of this species. Thus, it is important to maintain mature trees in addition to promoting seed production.     

华中樱桃种子生物学特性研究初探

为探究华中樱桃 Cerasus conradinae种子生物学特性,以 湖北省赤壁市的野生华中樱桃的自然成熟未落地的果实为试验 材料,观察其种子形态,并测定种子长度、宽度、厚度、千粒 重、吸水特性和发芽特性。结果表明：华中樱桃种子为卵形,长 7.64±0.73mm,宽5.28±0.55mm,厚4.39±0.47mm,千粒重 62.77±2.91g,属中粒种子;种子浸种0~4 h吸水较快,后渐趋 缓,浸种24 h近饱和,吸水率为33.23%,种子饱和状态时吸水率 为33.30%;用750 mg/L 赤霉素（GA3）溶液浸种处理的种子发芽 效果较清水浸种好,但差异不明显,两者发芽率分别为65.3%和 62.7%;华中樱桃幼苗个体间差异大。     

Pigment composition and location in honey locust (Gleditsia triacanthos) seeds before and after desiccation

Gleditsia triacanthos L. pods were harvested at San Lorenzo del Escorial, Madrid, Spain, before and after seed desiccation. Maturing green pods, harvested from trees, contained soft light-green seeds, whereas fully mature brown pods, collected on the ground, contained hard dark-green seeds. Based on visual inspection of seeds, it was determined that the green color is located in the cotyledons and embryo of soft seeds but mainly in the seed coat of hard seeds. High performance liquid chromatographic analysis indicated that both hard and soft seeds contained the same set of photosynthetic pigments as fully developed leaves, but in different proportions. The hard and soft seeds mainly differed in their chlorophyll a to chlorophyll b ratio and in the composition of the xanthophyll cycle pool of pigments. Fluorescence at -196 degrees C revealed that the molecular organization of the pigment molecules in the seed coat of hard seeds differed from that in intact cotyledons of soft seeds and intact green leaves. The -196 degrees C fluorescence spectra also revealed the presence of a small heterogenous pool of non-photoactive protochlorophyll(ide)s, similar to those found in dark-grown tissues of gymnosperms and angiosperms.     

Morphological and biochemical changes of Aesculus chinensis seeds in the process of maturation

Morphological and biochemical changes of Chinese horse chestnut (Aesculus chinensis Bge.) seeds were studied during the process of maturation. Fruit and seed diameter increased linearly from DPA (Days Post Anthesis) 60 to DPA 150 and were stable thereafter. The weight of fruit and seed increased sigmoidally between DPA 100 and DPA 160. The water contents of whole seeds, axes and cotyledons had a sharp reduction from DPA 110 to DPA 130 but changed little after DPA 130. The water contents of whole seeds, axes and cotyledons were 1.42, 1.93 and 1.30?g?g^?1DW, respectively at the end of maturation. The starch content of cotyledons from mature seeds was high on a dry weight basis (31.4%) while the total soluble sugar content varied between 12 and 20%. In both axes and cotyledons, the total soluble sugar content decreased during the process of maturation, as did all five soluble sugar components measured (glucose, sucrose, trehalose, raffinose and stachyose). Sucrose and stachyose contents remained relatively high at the time of seed maturation, followed by glucose and a trace level of trehalose. The oligosaccharide/disaccharide (O/D) ratio in the axes was 3.1 at DPA 140. The high level of sucrose and stachyose might contribute to the greater desiccation tolerance of Aesculus chinensis seeds relative to typical recalcitrant tropical seeds. High levels of ABA content in seed coats and a low GA₃/ABA ratio in both seed coats and axes might also influence the desiccation tolerance and shallow dormancy of mature Aesculus chinensis seeds.     

Plant regeneration by somatic embryogenesis from mature seeds of <Emphasis Type="Italic">Magnolia obovata</Emphasis>

We attempted to develop a method for the regeneration of plantlets from mature seeds of medically important Magnolia obovata via the induction of somatic embryogenesis in vitro. We initially cultured halves of mature seeds on either Murashige and Skoog (MS) medium or B₅ medium that contained 0, 1, 5 or 10 μM gibberellic acid (GA₃) for 1 month and then transferred the half-seeds to half-strength MS basal medium or B₅ basal medium for further culture in the absence of GA₃. Proembryogenic masses (PEMs) were observed 1 month after the transfer of the halved mature seeds to the medium without GA₃. The frequency of formation of PEMs was higher (28%) after initial culture in MS basal medium plus 1 μM GA₃ than in other tested media (0 or 4%). Somatic embryos that had been developed from PEMs were cultured on half-strength MS basal medium or B₅ basal medium for completion of maturation and then transferred to fresh aliquots of the same medium for initiation of germination. The frequency of germination, with the formation of normal primary leaves and roots, was above 80%. We transferred the somatic embryo-derived plantlets to soil for acclimatization and the plantlets continued to thrive.     

杉木种子发育生理、涩籽成因及降低败育措施的研究

对杉木种子发育生理及涩籽形成原因的研究结果表明：随着胚的分化发育,雌配子体（胚乳）和胚的干质量、体积及生物分子（总核酸、蛋白质和类脂）含量均不断增加,一直持续到种子成熟。败育的杉木涩籽中存在着凝集素类物质,它的存在与败育无关,而在贮藏蛋白的形成中的作用。败能的杉木涩籽中只积累较多的脱落酸,说明了败育涩籽的衰败和失活。根据研究结果提出了减少杉木种子败育的措施。     

Density-dependent selfing and its effects on seed performance in a tropical canopy tree species, Shorea acuminata (Dipterocarpaceae)

In the conservation and management practices of natural forests, sound reproduction and regeneration form the basis of the maintenance and viability of the tree populations. To obtain and serve biological information for sustainable forest management, we investigated reproductive biology and inbreeding depression in seeds of an important dipterocarp tree species, Shorea acuminata (Dipterocarpaceae), by both field and laboratory experiments. Results of parental analysis of immature and mature seeds showed that selfing rates varied greatly, from 7.6 to 88.4% among eight mother trees, and the mean overall selfing rate was 38.3%. Observed outcrossing events within a 40-ha study plot were predominantly (76.5%) short-distance events with a mating distance (md) ≤ 100 m. Since the selfing rate sharply decreased with increase in the number of flowering conspecifics (i.e., individuals of the same species) within a 100-m radius from the mother trees, the local density of flowering conspecifics appears to be the key factor determining the outcrossing rate in S. acuminata. However, the extremely high selfing rate (88.4%) observed for one tree could not be simply explained by the low local density of flowering conspecifics. Instead, differences in its flowering phenology (its flowering peaked ca. a week earlier than most of the other examined individuals) may have severely limited its receipt of pollen from other conspecifics, and thus promoted selfing. Since there were no significant differences in the proportion of selfed progeny between immature and mature seed stages, there was no evidence of selective abortion of selfed seeds during seed development. However, the seed mass of outcrossed progeny was heavier than that of selfed progeny, and heavier seeds showed higher success rates at germination and seedling establishment. These results suggest that inbreeding depression resulted in reductions in seed mass and may reduce the fitness of selfed seeds in S. acuminata. In addition, the outcrossing rate of S. acuminata was more sensitive to low local conspecific flowering-tree densities than that of a sympatric bee-pollinated dipterocarp species with greater pollination distances. These results suggest that the management of local adult-tree densities is important for avoiding selfing and inbreeding depression in future generations, especially in a species like S. acuminata with predominantly short-distance pollination.     

印加果种子发育过程中脂肪酸的累积规律

【目的】研究印加果种子中脂肪酸物质的累积规律,对印加果种子不同发育时期的脂肪酸物质进行解析,为印加果种植区域推广和新品种选育提供理论依据。【方法】分别选取成年印加果植株的18、36、54、72和90 DAF种子,利用液氮微量提取印加果种子的脂肪酸,通过气相色谱-质谱技术对其组分和相对含量进行动态变化的测定分析。【结果】印加果种子整个发育过程主要分离鉴定出56种物质,其中包括14种脂肪酸类化合物(80.13%),7种酯类化合物(2.70%),24种烷烃类化合物(12.40%),7种苯环类化合物(3.96%)以及4种其他化合物(0.81%)。脂肪酸类化合物主要包含亚油酸、亚麻酸、棕榈酸、硬脂酸、油酸及16-甲基十七烷酸等物质。随着成熟度增加,脂肪酸总含量逐渐从53.60%、76.30%、76.78%、96.43%增加至97.54%。其中,亚油酸和亚麻酸呈增加趋势,棕榈酸、硬脂酸、油酸及16-甲基十七烷酸等物质呈下降趋势。【结论】在印加果种子发育过程中,脂肪酸物质逐渐由饱和脂肪酸和单不饱和脂肪酸物质转化为多不饱和脂肪酸物质,相比其他油料植物具有成熟期短、不饱和脂肪酸转化效率高等优点,推测可能与其高效的光合速率相关。因此,印加果作为一种强需光性的食用油料植物,适宜在纬度较低的广东、广西、云南、福建、海南以及台湾等部分区域进行种植。     

Somatic embryogenesis from immature and mature zygotic embryos ofCryptomeria japonica I: Embryogenic cell induction and its morphological characteristics

Embryogenic cells (ECs) of sugi (Cryptomeria japonica D. Don) were induced from immature and mature zygotic embryos cultured on different concentrations of 2,4-dichlorophenoxyacetic acid (2,4-D)-containing modified Campbell and Durzan medium. The rate of induction of ECs varied depending on the stage of embryos collected. The highest percentage of induction (35%) was obtained with immature zygotic embryos collected July 18 and July 30, 1997, when 1 M of 2,4-D was added to the induction medium. The ECs easily proliferated when subcultured in a medium of the same composition as the induction medium within 3 weeks. Morphological characteristics of nonembryogenic cells and embryogenic cells of different developmental stages were studied under an inverted fluorescence microscope.Part of this work was presented at the 109th annual Japanese Forestry Society meeting at Tochigi, April 1998     

Desiccation and storage behavior of bay laurel (Laurus nobilis L.) seeds

The effect of seed moisture content (m.c.) and seed storage conditions of bay laurel (Laurus nobilis L.) was investigated in relation to seed viability. In the first experiment, the effect of drying rate on seed moisture and seed germination was investigated. Fresh seeds, with their original moisture content displayed a germination percentage of 55.1%. When the seed moisture content was reduced by 2.0% in an oven, the germination percentage rose to 81.0%. When the seed moisture content was reduced even more by using the same method, the germination percentages decreased dramatically. Reducing the seed moisture content to 28.7 and 23.5% by drying the seeds in alternating room conditions resulted in an increase of seed germinability to 84.3 and 90.9%, respectively. The drying of the seeds for 45, 60 and 75 days reduced their seed germination to 66.8, 49.4 and 48.0%, respectively. Reducing seed moisture content below 15.0% resulted in practically nullifying seed germinability. The fact that bay laurel seeds cannot retain their germinability at lower moisture contents demonstrates that it is a species with recalcitrant seeds. In the second experiment, moist and dry storage conditions were tested under different temperatures and moisture contents. The storage experiment showed that the most effective way of conserving the bay laurel seeds is moist storage at 0 ± 1°C for 4 months without previous drying of the seeds.     

Anthraquinone production by callus cultures of Rubia cordifolia

Munjistin and purpurin were identified as the major components of anthraquinone pigments produced by callus cultures of R. cordifolia. Anthraquinone content in calluses was 0.62–1.22% (by dry wt.) depending on the source of explants. Selection of coloured aggregates yielded a cell line with twofold increase in anthraquinone production.     

CELLULAR SUBSTANCE EXUDATION AND SEED VIGOUR

In this paper.seeds of pinus koraiensis which has a characteristic of deep dormancywere selected as an expriment material,and the relation of cellular substance exudation to seed vig-our is discussed based on the dormant state,the different measuring parts and the imbibed state ofseeds before testing,as well as on the respiration of embryo and endosperm.The major measured re-sults are as follows:(1)After soaking for 24h,the dry embryo conductivity of seedlot L is greaterthat than of seedlot H:(2)The substance exudation of dry embryo of seedlot H is less than that ofseedlot L;(3)After seeds imbibed,the substance exudation of dormant embryo of seedlot H is morethan that of seedlot L;(4)After germination hastening treatment,the substance exudation of H em-bryos is greater than that of L embryos;(5)After artificially accclcrated ageing treatment,the em-bryo conductivity of imbibed seeds has a positive correlation with seed vigour.