In vitro propagation of a medicinal plant: Tripterygium wilfordii Hook f.

In this study a reliable protocol was developed for the establishment of commercial in vitro cultures of Tripterygium wilfordii Hook f.. Juvenile shoots from one-year-old elite plants were used as the source of explants. New axillary shoots were obtained after 30 days of culture on a MS medium supplemented with BAP (2.0 mg·L^–1) and NAA (0.1 mg·L^–1). The optimal multiplication medium was a modified MS medium supplemented with BAP (1.0 mg·L^–1) and NAA (0.1 mg·L^–1). This yielded a multiplication rate of 2.4 for each subculture. Slightly more than 92% of shoots rooted when cultured on a modified MS medium containing IBA (0.2 mg·L^–1) and acti-vated charcoal (0.5 mg·L^–1). Activated charcoal promoted both a strong and a high rooting rate during the rooting phase. Plantlets were transferred to pots for a short acclimatization stage in a greenhouse where 95% of the plantlets survived. This highly reproduci-ble procedure can be adopted for large-scale propagation of T. wilfordii.     

马占相思优树组培快繁技术研究*

相思(Acacia Mill.)类树种(俗称相思树)属含羞草科(Mimosaceae)金合欢属(Acacia Mill.),在我国广东、海南、广西、福建、云南和江西等地均有栽培。相思树植株高大挺拔,根系发达具根瘤,具有耐干旱贫瘠及抗风的特性,是水土保持及改良土壤的优良树种;同时,相思树还是优良的用材树种,其生     

A study on the shade tolerance ofMuehlewbeckia complera

Muehlewbeckia complera was introduced to China in 2002 as indoor-hanging ornamental foliage plant. The experiment of the shade tolerance for this species was carried out in different light intensities (0.14–946.00 μmol·m⁻²·s⁻¹). After 40 days in experimental areas, leaf photosynthentic characteristics indexes ofM. complera in different photosynthesis active radiation (PAR) were measured with LI-COR6400 apparatus, such as the light compensation point, light saturation point, and maximum net photosynthesis rate, at the same time, the increments of total leaf area and leaf amount were measured. The results showed that the optimum light intensity range forM. complera was from 9.26 μmol·m⁻²·s⁻¹ to 569.00 μmol·m⁻²·s⁻¹ (463–28150 lx, relative humidity (RH) for 46–60%, temperature at 16–22°C). Under this condition, leaf photosynthetic efficiency was tiptop. AlthoughM. complera belonged to the moderate sun-adaptation plant species, the plant growth was inhibited when PAR increased to the level of 569.000 μmol·m⁻²·s⁻¹ or above.M. complera could sprout new leaves in photosynthesis active radiation of 0.16–19.22 μmol·m⁻²·s⁻¹ (8–961 lx), or 10 μmol·m⁻²·s⁻¹ for above 6 h. Foundation item: This study was supported by the Research Foundation of Northeast Forestry University. Biography: YUE Hua (1962-), female, Associate professor in Northeast Forestry University, Harbin 150040, P. R. China. Responsible editor: Zhu Hong     

Effects of Thidiazuron,basal medium and light quality on adventitious shoot regeneration from <Emphasis Type="Italic">in vitro</Emphasis> cultured stem of <Emphasis Type="Italic">Populus alba</Emphasis>×<Emphasis Type="Italic">P. berolinensis</Emphasis>

The effect of Thidiazuron （TDZ）, basal media and light quality on adventitious shoot regeneration from in vitro cultured stem of Populus albaxP berolinensis were determined to establish a high efficiency shoot regeneration system from stem explants of P. alba~P berolinensis. Stems ofPopulus alba~P berolinensis were collected from cultured shoots in vitro derived from dormancy buds of 3-year-old seedlings. The stem explants were cultured on MS medium containing 0.02-mg·L-1NAA （naphthaleneacetic acid）, and 0.1, 0.3, 0.5 and 1.0 mg·L-1 concentrations of TDZ to determine the effect of TDZ on shoot regeneration. Three basal media, i.e. MS, woody plant medium （WPM） and B5, were used to test their influences of different media on adventitious shoot regeneration. Green, red, blue and yellow plastic films in comparison with florescent light as control were used to observe their effects on shoot regeneration. The results showed that differ- ent concentrations of TDZ had an evident influence on shoot regeneration. Lower concentration of TDZ （0.1 mg·L-1） resulted in more ad- ventitious shoot regeneration and higher concentration of TDZ （〉0.1 mg·L-1） inhibited shoot regeneration. Among different media, MS medium exhibited a high efficiency for shoot regeneration, followed by WPM medium, while B5 medium inhibited shoot regeneration. Normal light and yellow light exhibited better effects on shoot regeneration, compared with other light.     

Effects of mother tree ages,different rooting mediums,light conditions and auxin treatments on rooting behaviour of <Emphasis Type="Italic">Dalbergia sissoo</Emphasis> branch cuttings

Dalbergia sissoo Roxb. is one of the promising multipurpose tree species of South Asia. Most of the plantations of D. sissoo from seeds are facing severe threats due to the die-back disease, which ultimately causes death of this potential tree-species within a few months. Vegetative propagation could avoid the die-back disease. Thirty mother trees of different age-groups of D. sissoo were selected for evaluating the rooting behaviour of branch cuttings from D. sissoo as influenced by auxins (IAA or IBA at 100, 200, 500 mg·L⁻¹), ages of mother trees (10, 4 and 2 years old) and different environment conditions, i.e., different mediums (soil and sand) or light conditions (in shade and open condition). The results show that application of IAA and IBA induced more numbers of cuttings (collected from 10-year-old mother trees) to root compared to control. Branch cuttings of D. sissoo collected from 10-year-old mother trees and planted in soil bed in open conditions had 100.0% of cuttings to root in IAA (100 mg·L⁻¹) and IBA (200 mg·L⁻¹) treatments. Both rooting medium (Soil and sand) influenced significantly (p<0.05) on rooting response of branch cuttings. Soil medium was found to achieve maximum no. of branch cuttings to root, compared to sand medium.     

Embryo culture and rapid propagation of Syringa

Embryo of lilacs (Syringa L) culture in vitro and the rapid propagation were studied. The orthogonal experiments, including the selection of basal medium, embryo age and other factors such as sugar, benzyladenine (BA), naphthalene acetic acid (NAA) and glutamine (Gln), were carried out. The results indicated that the optimal medium for embryo culture was Monnier medium supplemented with NAA (0.001 mg.L^-1), BA (0.1 mg.L^-1), sugar (50 g.L^-1), and Gin (400 mg.L^-1), with a germination rate of 91.7% at least; the optimal embryo age was 50 d; and Gln had significant effects on the germination rate of embryo.Moreover, the optimal medium for subculture was MS BA (2 mg.L^-1) NAA (0.001 mg.L^-1) Gln (0.5 mg.L^-1), with the propagation coefficient of 3.6 at least.     

Micropropagation of mature Terminalia catappa (Indian Almond), a medicinally important forest tree

We report an efficient in vitro propagation method for Terminalia catappa using nodal segments of a 15-year-old mature tree. The nodal segments were cultured on MS medium supplemented with 6-benzyladenine (BA; 0.5–3.0 mg l⁻¹) or Kinetin (Kn; 0.5–3.0 mg l⁻¹) for bud breaking and multiple shoot induction. About 85% of the explant responded (2.8 ± 0.41 shoots per node with 2.7 ± 0.14 cm length) within 15 days of inoculation in Murashige and Skoog medium fortified with 2.0 mg l⁻¹ of BA. Further shoot multiplication was achieved by repeated transfer of mother explants and subculturing of in vitro-produced shoots on medium supplemented with various concentrations of BA (0.25–1.5 mg l⁻¹) or Kn (0.25–1.5 mg l⁻¹) or on their combinations. Optimal number of shoots and shoot length were recorded on MS medium supplemented with 0.25 mg l⁻¹ of BA and 0.25 mg l⁻¹ of Kn. The multiplied shoots were used for ex vitro rooting after treatment for 4 min with indole-3-butyric acid (IBA; 50–500 mg l⁻¹) or α-naphthalene acetic acid (NAA; 50–500 mg l⁻¹). About 80% of the shoots treated with 200 mg l⁻¹ of IBA produced ex vitro roots with an average of 2.8 roots per shoot. Nearly 75% of these plantlets could be acclimatized within 5 weeks and successfully established in the field. This is the first report on micropropagation of T. catappa, which can be applied for further genetic transformation assays and pharmaceutical purposes.     

连香树嫩枝扦插繁殖技术研究

以连香树(Cercidiphyllum japonicum)当年生枝条为材料,研究了不同植物生长调节剂、不同IBA浓度及其处理时间对插穗生根的影响。结果表明:连香树属于皮部生根型,只有极少数为愈伤组织生根。植物生长调节剂处理是提高嫩枝插穗生根能力的一个必要措施,其中IBA对连香树插条生根效果最好,NAA次之,IAA最差。采用IBA 100 mg.L-1+NAA 100mg.L-1混合处理嫩枝插条效果最好,生根率达到81.67%,且幼苗长势良好。IBA 500 mg.L-1快蘸处理8 s,生根率达到71.67%;IBA 200 mg.L-1浸泡6 h,生根率为77.78%。综合各项生根指标,采用IBA 100 mg.L-1+NAA 100 mg.L-1浸泡嫩枝插条4 h,最适宜连香树插条生根。     

Micropropagation of <Emphasis Type="Italic">Salvadora</Emphasis><Emphasis Type="Italic">persica</Emphasis>-a tree of arid horticulture and forestry

A micropropagation method for Jaal (Salvadora persica)—a tree of arid horticulture and forestry has been developed. Nodal segments of fresh shoot sprouts originated from axillary buds obtained from a plant around 35–40 years old lopped plant were used as explants for establishment of in vitro cultures. Surface-sterilized explants produced optimum number of shoots through activation of axillary buds on Murashige and Skoog’s (MS) medium containing 8.88 μM BA (6-benzyladenine) + additives (25 mgl⁻¹ each of adenine sulphate, arginine, citric acid, 50 mgl⁻¹ ascorbic acid). The shoot multiplication was influenced by the successive transfer of the mother explants for 4–5 passages. The maximum number (23.1 ± 0.73 shoots per explant) of shoots were regenerated on MS supplemented with 1.11 μM BA + 1.16 μM Kn (Kinetin) + 0.54 μM NAA (α-naphthalene acetic acid). About 90% shoots pulse-treated with a combination of 2460.27 μM Indole-3-butyric acid (IBA) + 494.56 μM NOA (2-naphthoxy acetic acid) were rooted ex vitro on soilrite within 15–18 days. Over 80% cloned plantlets were hardened successfully in a green house and transferred to polybag/pots.     

东北刺人参组培快繁培养基的筛选

东北刺人参(Oplopanax elatusNakai.)又称刺参,五加科(Araliaceae)多年生落叶灌木,其干燥根和茎均可入药,是一种用途广泛的中药材。近些年来,医药科研工作者对其成分及药理进行了大量的试验和研究,其结果表明,根、茎入药有类似于人参的作用,可用于治疗神经衰弱、低血压和风湿性     

Establishment of an in vitro plant regeneration protocol for Casuarina cunninghamiana Miq. via indirect organogenesis

An in vitro plant regeneration protocol via indirect organogenesis from morphogenetic callus was established for Casuarina cunninghamiana Miq. Effects of plant growth regulator NAA (naphthaleneacetic acid) and BAP (6-benzylaminopurine), sucrose and AgNO₃ on callus induction, adventitious bud differentiation and shoot development were examined. Explants used were epicotyl fragments from 45-day-old seedlings. The largest callus (4.29 mm in diameter) was obtained after 1 month on a basic culture medium consisting of Murashige and Skoog ? macro- and full strength micro- elements, Nitsch and Nitsch vitamins, supplemented with 0.54 μM NAA, 3.30 μM BAP, and 30 g L⁻¹ sucrose. The calli were subcultured in the same medium above for 2 months. They were then cultured for another 2 months for adventitious bud differentiation and shoot development. The highest mean adventitious bud differentiation, number of shoots formed per callus and number of shoots ≥2 cm long per callus (47.50%, 27.38 and 4.75, respectively) were achieved on the above medium modified with NAA at 0.27 μM and supplemented with AgNO₃ 1 mg L⁻¹. Shoots were successfully rooted without plant growth regulator and the rooted plantlets survived and grew normally. This protocol for in vitro plant regeneration provides a tool not only for vegetative propagation but also for plant genetic transformation and gene function studies of C. cunninghamiana.     

余甘子离体快速繁殖技术的初步研究

余甘子 (ＰｈｙｌｌａｎｔｈｕｓｅｍｂｌｉｃａＬ .)为大戟科 (Ｅｕｐｈｏｒｂｉａｃｅａｅ) ,叶下珠属 (Ｐｈｙｌｌａｎｔｈｕｓ) ,落叶小乔木或灌木。在我国主要分布于南亚热带地区[1] 。余甘子果可生食、渍制或榨取果汁 ,果具清热解毒、降血压、防治肝胆病、收敛止泻作用 ,是重要的中药材和制药原料[2 ] ,同时树皮为重要的栲胶原料。余甘子在干热河谷、石质山地等生态脆弱区已作为优良的生态经济兼用造林树种。我国余甘子良种化起步晚 ,虽然只有少数地区开始采用良种 ,但发展势头很快。近几年引进的国外品种已表现出较好的适应性和生长…     

Photosynthetic induction responses of Pinus koraiensis seedlings grown in different light environments

The time processes of photosynthetic induction responses to various irradiances in Korean pine (Pinus koraiensis) seedlings grown in open-light environments and in understory of forest were studied in an area near the Research Station of Changbai Mountain Forest Ecosystems, Jilin Province, China from July 15 to August 5, 1997. The results showed that at 200 μmol·m⁻²·s⁻¹ photosynthetic photon flux density (PPFD) and 500 μmol·m⁻²·s⁻¹ PPFD, the induction time for the photosynthetic rates of understory-grown seedlings to reach 50% and 90% steady-state net photosynthetic rates was longer than that of the open-grown seedlings. The induction responses of open-growth seedlings at 500 μmol·m⁻²·s⁻¹ PPFD were slower than those at 200 μmol·m⁻²·s⁻¹ PPFD, but it was the very reverse for understory-growth seedlings, which indicates that the photosynthetic induction times of Korean pine seedlings grown in the understory depended on the sunfleck intensity. Biograph: ZHOU Yong-bin (1970-), female, associate professor of Shenyang Agricultural University, Shenyang 110161, P.R. China. Responsible editor: Song Funan     

Efficient in vitro regeneration of <Emphasis Type="Italic">Leucaena leucocephala</Emphasis> using immature zygotic embryos as explants

A method for efficient in vitro regeneration of Leucaena leucocephala cv. K636 was developed using immature zygotic embryos as the explant. Multiple shoot induction was achieved by culturing the explants on half-strength Murashige and Skoog (MS) medium supplemented with thidiazuron (TDZ) (0.03 to 1.5 μM). The maximum number of shoots per explants was achieved in 0.26 μM TDZ-supplemented media. TDZ concentration significantly influenced the induction of multiple shoots as well as the shoot-length. TDZ at 0.35 μM or higher concentrations resulted in abnormal stunted shoots. Full-strength MS media devoid of any plant growth regulator were used for shoot elongation. In vitro root induction was achieved in half-strength MS media supplemented either with 0.54 μM 1-naphthaleneacetic acid (NAA) or with 14.76 μM indole-3-butyric acid (IBA) in combination with 0.23 μM kinetin (Kn). Media supplemented with 14.76 μM IBA in combination with 0.23 μM Kn induced twofold–threefold more roots than media supplemented with 0.54 μM NAA. However, the average root length was lower in 14.76 μM IBA in combination with 0.23 μM Kn than in 0.54 μM NAA. Regenerated plants were maintained under normal condition after hardening. Plants, which were rooted on media supplemented with 14.76 μM IBA in combination with 0.23 μM Kn showed higher survival rate during hardening than those rooted on 0.54 μM NAA supplemented media.     

Composition and bioactivity of secretion from Stauronematus compressicornis (Fabricius)

We analyzed the composition and studied the bioactivity of secretion from the poplar sawfly, Stauronematus compressicornis (Fabricius). A leaf-sandwich method was applied to test the bioactivity of the secretion. Coomassie brilliant blue, gas chroma-tography (GC) and other methods were used to analyze and carry out protein assays, amino acid analyses, fatty acid and cholesterol detection of the secretion. Results show that the secretion had a significant antifeeding effect on 4th instar larvae of the armyworm (Pseudaletia separata Walker). Dissolved in 0.27 mol·L^–1 HCl, 0.1 mol·L^–1 NaOH and 0.05 mol·L^–1 Na₂HPO₄ separately, the secre-tions had antifeeding rates ranging from 22.7% to 49.6%. Meanwhile, the weights of the secretion-treated insects were significantly lower than those of the control. Biochemical detection showed that the proteins contained in the secretion were composed of 16 amino acids. The secretion also contained small amounts of cholesterol and fatty acids, which included lauric acid, myristic acid, myristoleic acid, seventeen carbonic acids and oleic acid.     

Variation in rooting behavior of stem cuttings in relation to their origin in Taxus wallichiana Zucc.

The present study investigates optimal conditions for the vegetative propagation of Himalayan yew Taxus wallichiana Zucc., an important medicinal tree, during spring. Effect of four treatments: (a) sex of donor plant (male and female), (b) age/type of shoot (1, 2, 3 year old, long and dwarf shoots), (c) auxin treatment (IBA and NAA at 0, 0.5, 1.25, 5.0 & 50.0 mM) and (d) rooting environment (raised beds/polythene bags) on percentage rooting in stem cuttings was studied. Randomized complete block (RBD) designs were used for experimentation. Rooting ability of cuttings was significantly influenced by all these treatments. The overall rooting response was higher in long shoot cuttings taken from female tree. Age of shoot also influenced the rooting response and was highest in 1 year old long shoot cuttings of female tree. Exogenous application of auxin, α-naphthalene acetic acid (NAA) and indole-3 butyric acid (IBA), had significant positive effect on the percentage rooting. IBA significantly enhanced the rooting percentage in 1 year old long and dwarf shoots at lower doses and 2 and 3 year old long shoots from female tree at higher doses. Maximum percent rooting (90% ± 2.8) was obtained with interactive effect of 0.5 mM, NAA (22 h) × 1 year old long shoot from female tree; followed by the interactive effect of 50 mM IBA (5 s) × 3 year old long shoot from female tree (83% ± 4.1). Cuttings planted in soil: sand medium in polythene bags showed earlier rooting response (12 weeks) than cuttings planted in raised nursery beds (24 weeks). Overall, the findings of this study suggest that 0.5 mM IBA treatment is suitable for enhancing adventitious rooting in 1 year old long and dwarf shoots of male and female trees. IBA at higher doses is suitable for enhancing the rooting percentage of 2 and 3 year old long shoots from female tree. This study provides a significant lead towards the development of a simple and inexpensive technique for large scale propagation, aforestation of elite genotypes and raising of bush type plantation under ex-situ conditions.     

An efficient micropropagation system for Celastrus paniculatus Willd.: a vulnerable medicinal plant

A micropropagation protocol was developed for Celastrus paniculatus, a vulnerable medicinal plant. Cultures were initiated from nodal explants collected from young shoots of a 12-year-old plant in MS basal medium. An average of five shoots were produced in MS medium supplemented with 1.5 mg l⁻¹ benzyl adenine (BA) and 0.1 mg l⁻¹ naphthalene acetic acid (NAA) after two subculture cycles with a 30-day interval. Continuous subculture in the same medium for three more cycles resulted in reduction of the number of multiple shoots (2 or 3 shoots), vitrification of the shoots, and callus formation. Vitrification of cultures could be overcome by the use of MS medium supplemented with lower concentrations of BA (0.05 mg l⁻¹) and NAA (0.01 mg l⁻¹). Among the various rooting trials, ex vitro rooting of shoots with simultaneous hardening was most efficient. The method standardized in the present study is simple, as it eliminated separate steps for in vitro rooting and hardening. Qualitative chemical similarity of the tissue culture regenerants with the mother plant was confirmed using high performance thin-layer chromatographic (HPTLC) profiling.     

Ecophysiological responses and carbon distribution ofPinus koraiensis seedlings to elevated carbon dioxide

The net CO₂ assimilation rate, stomatal conductance, RuBPcase (ribulose 1,5-biphosphate carboxylose) activity, dry weight of aboveground and belowgroud part, plant height, the length and diameter of taproot ofPinus koraiensis seedlings were measured and analyzed after six-week exposure to elevated CO₂ in an open-top chamber in Changbai Mountain of China from May to Oct. 1999. Seedlings were planted in four different conditions: on an open site, control chamber, 500 μL·L⁻¹ and 700 μL·L⁻¹ CO₂ chambers. The results showed that the total biomass of the seedlings increased whereas stomatal conductance decreased. The physiological responses and growth to 500 μL·L⁻¹ and 700 μL·L⁻¹ CO₂ varied greatly. The acclimation of photosynthesis was downward to 700 μL·L⁻¹ CO₂ but upward to 500 μL·L⁻¹ CO₂. The RuBPcase activity, chlorophyll and soluble sugar contents of the seedlings grown at 500 μL·L⁻¹ CO₂ were higher than that at 700 μL·L⁻¹ CO₂. The concentration 500 μL·L⁻¹ CO₂ enhanced the growth of aboveground part whereas 700 μL·L⁻¹ CO₂ allocated more carbon to belowground part. Elevated CO₂ changed the carbon distribution pattern. The ecophysiological responses were significantly different between plants grown under 500 μL·L⁻¹ CO₂ and 700 μL·L⁻¹ CO₂. Foundation Item: This paper was supported by Chinese Academy of Sciences. Biography: HAN Shi-jie (1956-), male, Ph. Doctor, Professor in Laboratory of Ecological Process of Trace Substance in Terrestrial Ecosystem, Institute of Applied Ecology, Chinese Academy of Sciences. Responsible editor: Chai Ruihai     

卷荚相思组培快繁技术研究

[目的]建立16年生卷荚相思优树组培快繁技术体系。[方法]以16年生卷荚相思优树的当年新生枝条带腋芽茎段为材料,对卷荚相思外植体进行消毒、初代培养、增殖培养、生根培养和移植等。[结果]表明:通过对16年生卷荚相思成年优树采条进行扦插,以扦插苗建立采穗圃,选取采穗圃中当年生健康无病虫害枝条的中段为外植体,最佳消毒方式为75%的酒精处理0.5 min和0.1%的升汞处理18 min,其存活率达69.33%,芽诱导率达86.67%;最佳初代培养基为改良MS+蔗糖40 g·L-1,出芽率为91.33%。最佳增殖培养基为MS+6-BA 0.5 mg·L-1+NAA 0.1 mg·L-1+蔗糖30 g·L-1,35 d增殖倍数可达3.50倍;最佳生根培养基为1/2 MS+IBA 0.25 mg·L-1+NAA 0.5 mg·L-1+蔗糖40 g·L-1,15 d生根率为96.11%;将生根苗移植至以沙为基质的营养杯中,存活率为71.11%。[结论]研究解决了16年生卷荚相思成年优树外植体污染率高和芽诱导率低等问题,建立的组培快繁技术体系对今后加快卷荚相思良种选育及优质苗木大量扩繁具有重要作用。     

Micropropagation and callus culture of <Emphasis Type="Italic">Saussurea laniceps</Emphasis>, an alpine medicinal plant

Cottonhead windhairdaisy (Saussurea laniceps Hand.-Mazz.) is one of the most famous and important medicinal herbs in China. Illegal collection from wild populations is increasingly threatening the present environment of S. laniceps. Establishment of an efficient method for micropropagation is the best way to change its endangered situation. When mature seeds of S. laniceps were cultured on hormone-free MS medium, plantlets were formed from germinated seeds in 7–10 d. Then 0.5 cm × 0.5 cm leaf explants were transplanted to MS medium supplemented with 1-naphthalene-acetic acid (NAA)/2,4-D and benzyladenine (BA)/KT and callus was achieved 10 d after transfer. Shoot bud regeneration occurred from callus cultured on MS medium supplemented with different growth regulators 20 d after culturing. The regeneration percentages varied with the different components of plant growth regulators. The percent regeneration from callus pretreated at low temperature of 5°C increased significantly compared with those incubated at 23/20°C directly. Optimal regeneration was observed with explants on media supplemented with 1.5 mg&#8226;L^–1 BA plus 0.2 mg&#8226;L^-1 NAA. In the presence of 0.2 mg&#8226;L^–1 NAA in half-strength MS, 78% of the shoots formed roots. Plantlets from explants showed 63% survival after acclimatization.