Effect of harvesting on temporal papyrus (<Emphasis Type="Italic">Cyperus papyrus</Emphasis>) biomass regeneration potential among swamps in Winam Gulf wetlands of Lake Victoria Basin,Kenya

This study established for the first time the impact of harvesting on post-harvest papyrus (Cyperus papyrus L.) biomass regeneration potential, with two harvesting regimes compared. Above-ground papyrus biomass was determined. Biomass varied with site. Site had no effect on regeneration potential, but monthly harvesting reduced papyrus biomass regeneration potential among sites. However, seasonal (6-monthly) harvesting did not appear to affect papyrus biomass regeneration potential. Exponential and polynomial trend analyses revealed a consistent downward trend for monthly harvest biomass, and the polynomial trend was more linear (F = 97.913; P < 0.001) than periodic (F = 9.617; P < 0.05). The polynomial trend scenario indicated how papyrus biological dynamics are likely to behave as monthly harvests are repeated. This suggests that regeneration potential is significantly reduced with successive monthly harvest, leading to weak spatial connectivity, papyrus stand fragmentation, and increased landscape patchiness. A 6-month harvest regime can be established to regenerate more biomass between harvests than is currently the case, with positive implications for wetland conservation and carbon sequestration. Papyrus harvesters can be kept off the swamps by establishing a riparian buffer zone of agro forestry trees and shrubs which can substitute for the papyrus as it is left to mature. However, while the information presented is useful for papyrus wetland management strategies, it is recognized that the study period was too short to permit a generalized recommendation.     

Management Impacts on Stand Structure of Lakeshore Phragmites Reeds

Many lakeside Phragmites australis reeds are frequently harvested or burnt in winter for several purposes like extraction of raw material, natural preservation or facilitation of professional fishing or fish breeding. The question is, whether these practices support a sustainable development of the reeds. Data from a long term monitoring program at Lake Constance (Germany) demonstrate that culm morphology, stand structure and aboveground peak biomass are strongly influenced by the treatment. Winter harvested and burnt reeds exhibited higher culm densities (+76% of the untreated controls), and a higher peak biomass (+13%). The mean culm height was reduced by −13%. The percentage of insect infested shoots was 8.8% of the total shoot population, compared with 20.1% of the untreated controls. Consecutive harvesting treatments intensified these effects. In the first two years after the treatment had stopped the reed stands showed an ‘overshoot’ biomass production, and a strong trend towards the features of the untreated controls. The possible mechanisms (mechanical damage, frost kill of the growth tips and breaking of the apical dominance) are discussed. It is concluded that winter harvesting and burning treatments make the reeds more susceptible against some environmental hazards, and should therefore applied with precaution on the basis of a close examination of risks and benefits.     

Primary productivity of papyrus (Cyperus papyrus) in a tropical swamp; Lake Naivasha,Kenya

Papyrus (Cyperus papyrus) standing biomass and the primary productivity of undisturbed and previously harvested areas of papyrus was measured in Lake Naivasha swamp, Kenya. Papyrus culm density in undisturbed swamp was estimated to be 13·1±1·9 culms m⁻² and aerial biomass was 3602 g m⁻². In undisturbed swamp the aerial productivity was 14·1 g m⁻² day⁻¹ while the previously harvested swamp reached a peak of 21·0 g m⁻² after 6 months. The annual aerial production rate of papyrus in Lake Naivasha was estimated to be 5150 g m⁻² year⁻¹. To sustain yields of regularly harvested papyrus swamps, the harvest intervals should exceed 1 year.     

Primary production in papyrus (<Emphasis Type="Italic">Cyperus papyrus</Emphasis> L.) of Rubondo Island,Lake Victoria,Tanzania

Above- and below-ground production and morphological characteristics of papyrus wetlands were measured at monthly intervals from July 2005 to June 2006 at Rubondo Island, Lake Victoria, Tanzania. The average value of live culm biomass (5,789 ± 435 g DW m⁻²) was higher than that of umbel biomass (2,902 ± 327 g DW m⁻²) by 50%. Root to rhizome means biomass value ratio was 1:1.8, rhizome biomass (4,144 ± 452 g DW m⁻²) being higher than roots biomass (2,254 ± 314 g DW m⁻²) by 45%. Direct proportion was observed between shoot density and culm–unit (culms and umbels) biomass. The average value of detritus/litter biomass (1,306 ± 315 g DW m⁻²) was less than total aerial biomass by 86%. The values of biomass are average of 12 sampling months from July 2005 to June 2006.     

Miscanthus×giganteus plant regeneration: effect of callus types,ages and culture methods on regeneration competence

The perennial rhizomatous grass, Miscanthus×giganteus is an ideal biomass crop due to its rapid vegetative growth and high biomass yield potential. As a naturally occurring sterile hybrid, M. ×giganteus must be propagated vegetatively by mechanically divided rhizomes or from micropropagated plantlets. Plant regeneration through somatic embryogenesis is a viable approach to achieve large‐scale production of plantlets in tissue culture. Effect of the callus types, ages and culture methods on the regeneration competence was studied to improve regeneration efficiency and shorten the period of tissue culture in M. ×giganteus. Shoot‐forming calli having a yellow or white compact callus with light‐green shoot‐like structures showed the highest regeneration frequency. Percentage of shoot‐forming callus induction from immature inflorescence explants was 41% on callus induction medium containing 13.6 μM 2,4‐d and 0.44 μM benzyladenine (BA). The use of a regeneration medium containing 1.3 μM NAA and 22 μM BA was effective at shortening the incubation period required for plantlet regeneration, with 69% of total regenerated plantlets obtained within 1 month of incubation on regeneration medium. Embryogenic‐like callus morphotype could maintain regeneration competency for up to 1 year as suspension cultures. Field grown regenerated plants showed normal phenotypic development with DNA content and plant heights comparable to rhizome propagated plants. Winter survival rates of the regenerated plants planted in 2006 and 2007 at the University of Illinois South Farm, Urbana‐Champaign, Illinois, were 78% and 56%, respectively.     

The potential use of papyrus (Cyperus papyrus L.) wetlands as a source of biomass energy for sub‐Saharan Africa

Four of five people in sub‐Saharan Africa rely on the traditional use of solid biomass, mainly fuelwood, for cooking. In some areas, the current rate of fuelwood consumption will exhaust biomass reserves within the next decade or two. A largely unrecognized source of biomass are tropical wetland ecosystems which have been shown to be some of the most productive ecosystems globally, exhibiting rates of net primary productivity comparable with high‐input, intensively managed agricultural systems. Papyrus (Cyperus papyrus L.) is an emergent sedge with C₄ photosynthesis which is native to the wetlands, river valleys and lakes of central, eastern and southern Africa. The mean standing dry matter of culms and umbels measured at a number of locations throughout East Africa is 38.3 ± 21.6 tDM ha^?1, and the aerial net primary productivity ranges between 25.9 and 136.4 tDM ha^?1 yr^?1. Papyrus vegetation can be harvested by hand and stacked on the rhizome mat for partial air‐drying, and it has been demonstrated that an annual harvesting regime has no negative impacts on long‐term productivity. The use of papyrus as a biofuel for cooking and heating depends on converting it to a suitably combustible form, such as compressed or carbonized briquettes with a calorific value approximately one‐third less than wood charcoal. While papyrus has significant potential as a biofuel, we argue that an integrated management and decision‐making framework for the sustainable utilization of papyrus wetlands is required, in which all ecosystem services including the provision of biomass energy need to be assessed. Sustainability of papyrus wetlands requires management which combines the strength of traditional communal governance and modern legislation to promote its utilization. In this way, local communities can benefit from the inherent advantages of tropical wetlands as very productive ecosystems.     

Phenology, starch allocation, and environmental effects on Myriophyllum aquaticum

Seasonal biomass and starch allocation patterns were determined from natural populations of Myriophyllum aquaticum that were sampled monthly from January 2006 to December 2007 in Mississippi. Water temperature, water depth, light irradiance, light transmittance, pH, and conductivity were also recorded during biomass harvests. Overall, few significant relationships were observed between the environmental factors tested and seasonal biomass. Submersed shoot biomass was negatively related (p < 0.01) with water temperature. Stolons accounted for 40–95% of total biomass followed by emergent shoot, submersed shoot, and root biomass. Percent starch in plant tissues was positively related to water temperature. Starch allocation was greatest in stolons where up to 16.3% of total starch was stored. Submersed shoots stored 0.6–11.0% of total starch followed by emergent shoots (0.4–7%). The roots of M. aquaticum stored less than 3.8% of total starch throughout the study period. Reduced biomass and starch storage occurred from October to March in both 2006 and 2007. Management strategies for this species could utilize an integrated approach to exploit times of low energy reserves (fall and winter), or to remove emergent shoots to gain access to the stolons and other submersed tissues.     

<Emphasis Type="Italic">In vitro</Emphasis> propagation of northern red oak (<Emphasis Type="Italic">Quercus rubra</Emphasis> L.)

In vitro propagation of northern red oak (Quercus rubra) shoots was successful from cotyledonary node explants excised from 8-wk-old in vitro grown seedlings. Initially, four shoots per explant were obtained on Murashige and Skoog (MS) medium supplemented with 4.4 μM 6-benzylaminopurine (BA), 0.45 μM thidiazuron (TDZ), and 500 mg l⁻¹ casein hydrolysate (CH) with a regeneration frequency of 64.7% after 3 wk. Subculturing explants (after harvesting shoots) to fresh treatment medium significantly increased shoot bud regeneration (16.6 buds per explant), but the buds failed to develop into shoots. A higher percentage (73.3%) of the explants regenerated four shoots per explant on woody plant medium (WPM) supplemented with 4.4 μM BA, 0.29 μM gibberellic acid (GA₃), and 500 mg l⁻¹ CH after 3 wk. Explants subcultured to fresh treatment medium after harvesting shoots significantly increased shoot regeneration (16 shoots per explant). Shoot elongation was achieved (4 cm) when shoots were excised and cultured on WPM supplemented with 0.44 μM BA and 0.29 μM GA₃. In vitro regenerated shoots were rooted on WPM supplemented with 4.9 μM indole-3-butyric acid. A higher percentage regeneration response and shoot numbers per explant were recorded on WPM supplemented with BA and GA₃, than on MS medium containing BA and TDZ. Lower concentrations of BA and GA₃ were required for shoot elongation and prevention of shoot tip necrosis. Each cotyledonary node yielded approximately 20 shoots within 12 wk. Rooted plantlets were successfully acclimatized.     

Micropropagation through excised root culture of Clitoria ternatea and comparison between in vitro–regenerated plants and seedlings

An efficient protocol has been developed for high‐frequency shoot regeneration and plant establishment of Clitoria ternatea – a potential medicinal legume. Adventitious shoots were regenerated from young excised root segments of aseptic seedlings on Murashige and Skoog (MS) medium supplemented with various concentrations of 6‐benzyladenine (BA), kinetin, α‐naphthalene acetic acid (NAA) or 2,4‐dichlorophenoxy acetic acid (2,4‐D) either singly or in various combinations. The highest frequency (100%) of shoot regeneration and maximum number (16.4 ± 0.24) of shoots per explant was obtained on MS medium supplemented with 20 μm BA and 2.0 μm NAA. Organogenic calli were produced on a medium containing 2,4‐D (10 or 20 μm ) and BA (5.0 μm ). The calli were differentiated into multiple shoots on MS medium supplemented with 2.5–10 μm BA and 2.0 μm NAA. The microshoots were rooted on half‐strength MS medium supplemented with 5.0 μm indole‐3‐butyric acid and transplanted successfully in field conditions. After 12 months of transfer to ex vitro conditions, the performance of micropropagated plants were evaluated on the basis of some physiological and biochemical parameters and compared with the in vivo–grown plants of the same age. The sodium dodecyl sulphate polyacrylamide gel electrophoresis protein profile was same between regenerated and naturally growing shoots. Total soluble protein in aerial part as well as in seeds of in vitro–regenerated and in vivo–grown plants was almost the same. The mitotic study showed normal chromosomal movement and numbers (2x = 16).     

Factors influencing plant regeneration from seedling explants of Hairy nightshade (<Emphasis Type="Italic">Solanum sarrachoides</Emphasis>)

Hairy nightshade (Solanum sarrachoides) has the potential to be a model system for the study of plant-pathogen interactions, however, the availability of tissue culture and transformation methods would strengthen its utility. For the development of tissue culture methods, we investigated, explant type (cotyledons, hypocotyls, roots), hypocotyl explant origin, cotyledon orientation (abaxial vs. adaxial) in direct contact with the medium, gelling agents (agar and agargel) and cytokinins (zeatin and 6-benzyladenine) at different concentrations. Cotyledon explants resulted in the greatest biomass as compared to root and hypocotyl. As for hypocotyl explant origin, explants proximal to the cotyledons had a significant effect on plant regeneration. However, cotyledon orientation and gelling agent had no effect on plant regeneration. Medium supplemented with either zeatin or 6-benzyladenine at 1 mg L⁻¹ resulted in significant shoot regeneration. Shoots rooted readily when cultured on a non-hormone based rooting medium.     

Light capture efficiency decreases with increasing tree age and size in the southern hemisphere gymnosperm Agathis australis

We investigated leaf and shoot architecture in relation to growth irradiance (Q_int) in young and mature trees of a New Zealand native gymnosperm Agathis australis (D. Don) Lindl. to determine tree size-dependent and age-dependent controls on light interception efficiency. A binomial 3-D turbid medium model was constructed to distinguish between differences in shoot light interception efficiency due to variations in leaf area density, angular distribution and leaf aggregation. Because of the positive effect of light on leaf dry mass per area (M_A), nitrogen content per area (N_A) increased with increasing irradiance in both young and mature trees. At a common irradiance, N_A, M_A and the components of M_A, density and thickness, were larger in mature trees, indicating a greater accumulation of photosynthetic biomass per unit area, but also a larger fraction of support biomass in older trees. In both young and mature trees, shoot inclination angle relative to horizontal, and leaf number per unit stem length decreased, and silhouette to total leaf area ratio (S_S) increased with decreasing irradiance, demonstrating more efficient light harvesting in low light. The shoots of young trees were more horizontal and less densely leafed with a larger S_S than those of mature trees, signifying greater light interception efficiency in young plants. Superior light harvesting in young trees resulted from more planar leaf arrangement and less clumped foliage. These results suggest that the age-dependent and/or size-dependent decreases in stand productivity may partly result from reduced light interception efficiency in larger mature relative to smaller and younger plants.     

Moderate desiccation dramatically improves shoot regeneration from maize (<Emphasis Type="Italic">Zea mays L.</Emphasis>) callus

Tissue and cell culture systems are vital to many areas of maize research and improvement. Efficient shoot regeneration remains a limiting factor for most elite lines. To enhance shoot regeneration, calluses derived from immature embryos of four genotypes were subjected to 6, 12, 24, 48, and 72 h of desiccation on sterile filter paper before shoot induction. We achieved up to 32% desiccation (measured as mass) and 48% imbibition (measured as mass gain in 48 h incubation) which was 3.4-fold higher than nondesiccated controls. This rapid desiccation/imbibition procedure enhanced shoot regeneration and development, but its efficiency was crucially dependent on the degree and duration of desiccation. All four genotypes calluses regenerated shoots best after 48 h of desiccation corresponding to a 23.7% average desiccation percentage, and regeneration frequencies reached 42–74%, which increased 1.5–2.1-fold compared with that of nondesiccated calluses. The effect of desiccation treatment on the shoot regeneration increase took place mainly during the early phase of induction. The weekly average shoot regeneration frequency of calluses with 48 h desiccation in the first wk reached 8.8% (the control was no regeneration); then it increased 3.0 and 2.3-fold in the second and third wk induction, respectively. We demonstrate that the moderate desiccation of calluses increases the yield of shoot regeneration and speeds up regeneration course in maize.     

The effects of genotype,inflorescence developmental stage and induction medium on callus induction and plant regeneration in two <Emphasis Type="Italic">Miscanthus</Emphasis> species

Several grass species of the genus Miscanthus are considered to be outstanding candidates for a sustainable production of biomass to generate renewable energy. The purpose of this study was to investigate the effects of genotype, the developmental stage of the explant donor inflorescence and the induction medium on the success rate of micropropagation. The experiments were conducted on three genotypes of M. sinensis and one of M. x giganteus. Explants from the youngest inflorescences (0.1–2.5 cm in length) showed a significantly higher callus induction rate than those from more developed inflorescences (2.6–5 cm in length). In addition, cultures initiated from explants from the youngest inflorescences showed significantly the highest rates of callus regeneration and the highest shoot regeneration rate. Three out of the four genotypes tested showed the best shoot regeneration from calli initiated from the youngest inflorescences when cultured on the Murashige and Skoog basal medium (MS) with 5 mg l⁻¹ 2,4-dichlorophenoxyacetic acid (2,4-D) and 0.1 mg l⁻¹ 6-benzyladenine (BA). The percentages of calli from those genotypes showing regeneration ranged from 45 to 76.7%, and the corresponding shoot regeneration rates ranged from 1.85 to 6.33 shoots/callus. This demonstrates that, with some adjustments, efficient micropropagation of Miscanthus sp. is feasible.     

苦竹-杉木混交林界面区克隆分株秆形和地上生物量分配的适应策略

苦竹(Pleioblastus amarus)是优质笋材兼用竹种,分布广。为探究界面区苦竹分株秆形及地上构件生物量分配格局的变化特征,解析苦竹对异质生境适应机制,该研究选取了相邻的苦竹林和苦竹-杉木(Cunninghamia lanceolata)混交林两种林分类型,分别测定了苦竹林和混交林中心区及界面区不同龄级立竹秆形和秆、枝、叶的生物量,分析立竹秆形及地上构件生物量积累、分配、异速生长关系的差异。结果表明:(1)界面区1 a立竹生物量积累及分配差异增大,其中苦竹林界面区各构件相对生物量和叶生物量分配比例提高,而混交林界面区各构件相对生物量和叶生物量分配比例降低; 2 a立竹生物量积累及分配比例的差异缩小,界面区两边2 a立竹各构件相对生物量和生物量分配比例均无明显差异。(2)界面区立竹秆形特征及1 a立竹各构件生物量异速生长关系均无明显变化,而苦竹林界面区2 a立竹秆的增长速率提高,枝、叶的增长速率降低。综上认为,苦竹通过权衡资源分配关系,明显改变界面区立竹秆形及生物量分配格局,以提高克隆分株对异质环境的适合度。     

Phenotypic plasticity of <Emphasis Type="Italic">Spartina alterniflora</Emphasis> and <Emphasis Type="Italic">Phragmites australis</Emphasis> in response to nitrogen addition and intraspecific competition

Phenotypic plasticity of the two salt marsh grasses Spartina alterniflora and Phragmites australis in salt marshes is crucial to their invasive ability, but the importance of phenotypic plasticity, nitrogen levels, and intraspecific competition to the success of the two species is unclear at present. Spartina alterniflora Loisel. is an extensively invasive species that has increased dramatically in distribution and abundance on the Chinese and European coasts, and has had considerable ecological impacts in the regions where it has established. Meanwhile, Phragmites australis Cav., a native salt marsh species on the east coast of China, has replaced the native S. alterniflora in many marshes along the Atlantic Coast of the US. This study determined the effects of nitrogen availability and culm density on the morphology, growth, and biomass allocation traits of Spartina alterniflora and Phragmites australis. A large number of morphological, growth, and biomass parameters were measured, and various derived values (culm: root ratio, specific leaf area, etc.) were calculated, along with an index of phenotypic plasticity. Nitrogen addition significantly affected growth performance and biomass allocation traits of Spartina alterniflora, and culm density significantly affected morphological characteristics in a negative way, especially for Spartina alterniflora. However, there were no significant interactions between nitrogen levels and culm density on the morphological parameters, growth performances parameters, and biomass allocation parameters of the two species. Spartina alterniflora appears to respond more strongly to nitrogen than to culm density and this pattern of phenotypic plasticity appears to offer an expedition for successful invasion and displacement of Phramites australias in China. The implication of this study is that, in response to the environmental changes that are increasing nitrogen levels, the range of Spartina alterniflora is expected to continue to expand on the east coast of China.     

Above- and below-ground competition between the liana <Emphasis Type="Italic">Acacia kamerunensis</Emphasis> and tree seedlings in contrasting light environments

Proliferation of lianas in canopy gaps can restrict tree regeneration in tropical forests through competition. Liana effects may differ between tree species, depending on tree requirements for above- and below-ground resources. We conducted an experiment in a shade house over 12 months to test the effect of light (7 and 27% external irradiance) on the competitive interactions between seedlings of one liana species and three tree species and the contribution of both above- and below-ground competition. Seedlings of the liana Acacia kamerunensis were grown with tree seedlings differing in shade tolerance: Nauclea diderrichii (Pioneer), Khaya anthotheca (Non-Pioneer Light Demander) and Garcinia afzelii (Non-Pioneer Shade Bearer). Trees were grown in four competition treatments with the liana: no competition, root competition, shoot competition and root and shoot competition. Both root and root–shoot competition significantly reduced relative growth rates in all three tree species. After one year, root–shoot competition reduced growth in biomass to 58% of those (all species) grown in no competition. The root competition treatment had a more important contribution in the effect of the liana on tree growth. Tree seedlings did not respond to competition with the liana by altering their patterns of biomass allocation. Although irradiance had a great effect on tree growth and allocation of biomass, the interaction between competition treatments and irradiance was not significant. Nauclea diderrichii, the tree species which responded most to the effects of competition, showed signs of being pot-bound, the stress of which may have augmented the competition effects. The understanding of the interaction of above- and below-ground competition between lianas and trees and its moderation by the light environment is important for a proper appreciation of the influence of lianas on tropical forest regeneration.     

Stand and self-thinning dynamics in natural <Emphasis Type="Italic">Abies</Emphasis> stands in northern Hokkaido,Japan

Stand dynamics and self-thinning were analyzed in relation to the dynamics of above-ground biomass in natural Abies sachalinensis stands growing on sand dunes in northern Hokkaido, Japan. This was done in order to examine wave-type regeneration in the stands. Fifty-two plots were established in almost pure Abies stands that ranged from saplings to the mature and collapsing growth stages. Above-ground biomass and tree height reached asymptotic levels prior to the collapsing phase, unlike wave-regeneration Abies stands in central Japan and North America. Stand density was high in the young growth stages, but the self-thinning rate, that is, the density decrease per biomass growth in the study stands was greater than in wave-regeneration stands in central Japan, as indicated by a large self-thinning exponent (–1.26 by reduced major axis regression). The range of tree height distribution was very narrow, and the stands vertical structure was typically single-layered. The slenderness ratio of trees was large, except in young stands. In mature and collapsing stands, advanced seedling density increased markedly. These stand and tree characteristics were considered to be correlated with the wave-type regeneration in the study stands, and it is assumed that prevailing winds affect tree mortality.     

模拟增温和降水变化对北京东灵山辽东栎种子出苗和幼苗生长的影响

气候变化将增加地表平均气温、改变降水格局, 会影响到种子出苗和幼苗生长, 进而影响物种的更新动态。为探讨增温和降水变化对东灵山地区建群树种辽东栎(Quercus mongolica)种子出苗和一年生幼苗生长和适应状况的影响, 该文利用环境控制生长箱开展了温度和降水量的双因素控制实验, 温度设置3个梯度: 月平均气温(对照)、增温2 ℃和增温6 ℃; 降水量设置3个梯度: 月平均降水量(对照)、减水30%和加水30%。结果表明: 1)辽东栎的种子出苗率和一年生幼苗的生长对增温和降水变化的响应不一致, 种子出苗率主要受到降水及其与温度交互作用的影响, 幼苗生长仅受到温度和降水独立作用的影响; 2)春季增温2 ℃或降水量增加均使辽东栎种子出苗期提前; 增温6 ℃与降水量减少的水热组合延迟了种子出苗期并使其存活率和出苗率显著降低, 但在此温度下增加降水量则增加了出苗速率和出苗率。3)增温2 ℃对其生长无显著影响, 增温6 ℃则在不同水分条件下显著地增加了幼苗的比叶面积、抑制了叶的伸长生长, 同时也显著降低了各器官生物量积累, 并减少了幼苗生物量向根的分配; 降水量减少降低了幼苗根生物量, 但未影响总生物量和根冠比, 降水量增加显著促进了幼苗地上部分的生长, 特别是叶的生长。因此, 适当地增温或增加降水量将增加辽东栎幼苗的更新潜力, 但增温和降水量减少导致的干旱化将显著降低幼苗的更新潜力。     

<Emphasis Type="Italic">In vitro</Emphasis> regeneration of <Emphasis Type="Italic">Echinacea pallida</Emphasis> from leaf explants

Summary A method has been developed for the induction of adventitious shoots from leaf tissue of Echinacea pallida with subsequent whole-plant regeneration. Proliferating callus and shoot cultures were derived from leaf tissue explants placed on Murashige and Skoog medium supplemented with 6-benzylaminopurine and naphthaleneacetic acid combinations. The optimum shoot regeneration frequency (63%) and number of shoots per explant (2.3 shoots per explant) was achieved using media supplemented with 26.6 μM 6-benzylaminopurine and 0.11 μM naphthaleneacetic acid. Rooting of regenerated shoot explants was successful on Murashige and Skoog medium, both with and without the addition of indole-3-butyric acid. All plantlets survived acclimatization, producing phenotypically normal plants in the greenhouse. This study demonstrates that leaf tissue of E. pallida is competent for adventitious shoot regeneration and establishes a useful method for the micropropagation of this important medicinal plant.     

Direct shoot regeneration from leaf explants of <Emphasis Type="Italic">Rosa damascena</Emphasis> Mill.

Summary A protocol for in vitro propagation using direct induction of shoot buds from leaf explants of in vitro-raised shoots of Rosa damascena var. Jwala is reported. The present study is the first report on direct shoot regeneration in scented roses. Elite plants raised from nodal explants and maintained for over 2yr in vitro on a static liquid shoot multiplication Murashige and Skoog (MS) medium supplemented with 5.0 μM benzyladenine (BA) and 3% sucrose were used. Petioles from fully developed young leaves, obtained after 4 wk of pruning of old shoots, were found to be ideal for regeneration of shoots. Initially the explants were cultured in an induction medium [half-strength MS+3% sucrose+6.8μM thidiazuron+0.27 μM α-naphthaleneacetic acid (NAA)+17.7 μM AgNO₃] and subsequently transferred to the regeneration medium (MS+2.25 μM BA+0.054 μM NAA) after 7, 14, 21, 28, and 35d. The highest shoot regeneration response (69%) was recorded when shoots were kept in the induction medium for 21 d and later transferred to regeneration medium. Histological studies revealed direct formation of shoot buds without the intervening callus phase. In vitro rooting of micro-shoots was accomplished within 2wk on half-strength MS liquid medium supplemented with 10.0 μM IBA and 3% sucrose for 1 wk in the dark and later transferred to hormone-free medium and kept in the light. Plantlets, remaining in the latter medium for 5–6 wk when transferred to soil, showed 90% survival.