An efficient method for clonal propagation and in vitro establishment of softwood shoots from epicormic buds of teak (Tectona grandis L.)

Softwood shoots were produced from 40 cm long stem segments placed horizontally in flat trays containing sterilized sand under natural light or shade conditions for subsequent rooting and micropropagation studies in teak (Tectona grandis L.). Higher number of shoots (6.17) per log was produced under natural light as compared to shade conditions. Forcing was also better in natural light as compared to shade in terms of shoot length, number of nodes or leaves. For rooting, 2–4 cm long softwood shoots were excised and treated with either indole-3-butyric acid (IBA) or α-naphthyl acetic acid (NAA) at 0, 1000, 2000 or 3000 μmol·L^–1 each or with combinations (1000 + 1000, 2000 + 2000 or 3000 + 3000 μmol·L^–1) and then placed in flat trays containing autoclaved sand at 25 ± 2ºC in 16 h photoperiod at 35 µmol·m^–2·s^–1. After 28 days, softwood cuttings treated with IBA + NAA (3000 + 3000 μmol·L–1) had highest rooting percentage (89.3%) with 5.5 mean roots. Shoot apex and nodal explants of softwood cuttings were pretreated with 0.1% (w/v) ascorbic acid, boric acid, activated charcoal, citric acid, glutamine or polyvinylpolypyrollidone (PVP) for 24 h to remove phenolic compounds before surface disinfestation. Glutamine (Gl) and PVP were equally effective resulting in 60% establishment of shoot apices on MS medium supplemented with 10 μmol·L–1 6-benzylaminopurine (BAP) + 5 μmol·L–1 NAA. Using shoot apices, highest (42.80) number of multiple shoots with 54.33 mm shoot length were obtained on MS + BAP (8.8 μmol·L–1) + IBA (2 μmol·L–1) after 45 days. Shoots were successfully rooted and acclimatized to greenhouse     

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.     

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.     

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.     

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     

Somatic embryogenesis and histological analysis from zygotic embryos in Vitis vinifera L. ＇Moldova＇

We examined the somatic embryogenesis from and histological studies of zygotic embryos of seeds in European Grape ＇Moldova＇ （Vitis vinifera U ＇Moldova＇）. Primary calli were initiated on Nitsch and Nitsch （NN） medium supplemented with 1.0 mg·L^-1 2,4-D and 0.5 mg·L^-1 6-BA. Embryogenic calli were produced upon transfer to a NN medium with 0.5 mg·L^-1 6-BA and 2 mg·L^-1 NAA and somatic embryos were obtained on a half strength MS medium without plant growth regulators. During the somatic embryo germination, an addition of 1.0 mg·L^-1 6-BA in the medium could accelerate somatic embryos to develop into normal plants and increase the conversion rate from 0 to 43.3%. Histological studies of embryogenic calli and somatic embryos demonstrated dynamic changes of proteins and starch grains. The developmental processes of somatic embryos were similar to those of zygotic embryos, including typical epiderma, cotyledon primordium and vascular tissue.     

Establishment of cell suspension line of <Emphasis Type="Italic">Populus tomentosa</Emphasis> Carr.

Leaves of fine Populus tomentosa genotype TC152 were used as explants to establish cell suspension lines. The effects of plant growth regulators on callus induction and establishment of cell suspension lines were studied. The callus induction rate was the highest on a MS solid medium supplemented with 1.0 mg·L^-1 2,4-D. A cell suspension line could be obtained by inoculating calli which were not subcultured into a MS liquid medium supplemented with 1.5 mg·L^-1 2,4-D. The best subculture medium was MS ＋ 0.8 mg＇L-1 2,4-D ＋ 30 g·L^-1 sucrose with a subculture cycle of seven days.     

Regeneration of <Emphasis Type="Italic">Melia volkensii</Emphasis> Gürke (Meliaceae) through direct somatic embryogenesis

Experiments were conducted to study plant regeneration through direct somatic embryogenesis using mature zygotic embryo and cotyledonary explants from seeds of Melia volkensii stored for <3 and >12 months. Explants were cultured on Murashige and Skoog (MS) medium supplemented with BAP, NAA and 2,4-D (0.5, 1.0 and 2.0 mg l⁻¹) alone, and BAP (0.5, 1.0, 2.0 and 4.0 mg l⁻¹) in combination with 2,4-D or NAA (0.2 and 0.5 mg l⁻¹). After 4 weeks in culture, up to 60% of cotyledonary explants from the seeds stored for <3 months produced direct somatic embryos on BAP (0.5–4.0 mg l⁻¹) in combination with 2,4-D (0.2 mg l⁻¹). The number of somatic embryos ranged from 5 to 14 per explant in BAP (0.5 mg l⁻¹) and 2,4-D (0.2 mg l⁻¹) combination. Only 20% of cotyledonary explants from seeds stored for >12 months produced somatic embryos. Mature zygotic embryos failed to produce any somatic embryos. Subcultures of somatic embryos from cotyledonary explants of seeds stored for <3 months formed clusters of shootlets on semi solid MS and 1/2 MS media. After 6 weeks of subculture on multiplication MS media augmented with BAP (0.5 mg l⁻¹) and IAA (0.2 mg l⁻¹), 70% of the shoot tips formed 4–7 shoots per explant. Up to 33% of the multiplied shoots were rooted in MS medium supplemented with 2.0 mg l⁻¹ IBA. Plantlets developed normally into seedlings in the greenhouse.     

美国枫香茎段组织培养与植株再生

美国枫香(Liquidambar styraciflua L.)为金缕梅科(Hamanelidaceae)枫香属(liquidambar L.)落叶乔木。生长迅速,树高18～20m,夏末叶片变为红、黄、紫等多种混合颜色,红叶期长,喜阳,适合酸性至中性土壤,广泛分布于北美南部以及墨西哥、中美洲至洪都拉斯的高海拔山区,是胶合板和造纸的良好用材^[1],也是良好的造林绿化树种,很有发展前景。     

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…     

白花泡桐优树试管嫁接幼化及组培快繁技术研究

以白花泡桐优树‘白优2号’为试验材料,通过组织培养和试管嫁接方法,对白花泡桐优树材料的幼化技术进行了研究.结果表明:外植体初代培养萌发的嫩芽为最适合的接穗;‘建始桐3号’为试管嫁接较合适的砧木;采用劈接进行;MS+ NAA0.3 mg·L-1+蔗糖30 g·L-1为试管嫁接培养基;继代增殖和生根培养基分别为1/2MS ...     

Establishment of genetic transformation system via Agrobacterium in tall fescue cultivar

Tall fescue (Festuca arundinacea Schreb.) is a cool-season turfgrass used on fairways in golf courses. The object of this study was to develop a more efficient, reliable, and repeatable approach in transforming the grass using Agrobacterium (EHA105), where β-glucuronidase gene (uidA) was used as a reporter and hygromycin phosphotransferase gene (hyg) as a selectable marker. An effective expression of transgene was observed in transforming 2-month-old calli derived from mature seeds (cv. Bingo) cultured on MS medium supplemented with 9 mg·L⁻¹ 2, 4-D. A two-step solid medium selection with increasing hygromycin concentration (from 30 to 50 mg·L⁻¹) was used to obtain resistant calli. Transgenic plants have been produced from many independent transformed calli. The presence of functional β-glucuronidase gene (uidA) was detected in hygromycin-resistant calli. Transgenic plants were regenerated and PCR and Southern blot confirmed transgene integration in the tall fescue genome. Biography: QIAN Hai-feng (1973–), male, Ph.D., associate professor in Zhejiang University of Technology.     

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.     

孟加拉哈提亚地区海岸造林对土壤性质的影响

在孟加拉诺阿卡利地区及相临裸地,对海岸植被(12年和17年生无瓣海桑Sonneratia apetala)进行探索性研究,以便了解海岸造林对土壤特性的影响.在三种不同地带(内陆、中部、海边),在12年生和17年生无瓣海桑林下,土壤深度分别为0-10,10-30和30-40cm,土壤湿度、土壤粒度、有机质、C含量、总N、pH、有效P、K、Na、Ca和Mg含量明显高(p≤0.05,p≤0.01,p≤0.001)于其相临裸地的数据,土壤含盐量明显(p≤0.01)低于其相临裸地的数据.在内陆CharAlim植被,土壤表面的土壤湿度,土壤粒度,有机质,C含量、总N、pH、土壤含盐量、有效P、K、Na、Ca和Mg含量分别为:31.09%、2.24 g·cm-3、2.41%、4.14%、0.58%、7.07、O.09 dS·cm-1、28.06 mg·L-1、O.50 mg·L-1、11.5 mg·L-1、3.30 mg·L-1和2.7 mmol·kg-1;而在相邻的Char Rehania贫瘠地区的相同土壤深度,其相关值分别为:16.69%、1.25g·cm-3、O.43%、0.74%、O.25%、6.57、0.13 dS·cm-1、13.07mg·L-1、O.30mg·L-1、1.4 mg·L-1、O.30 mmol·kG-1和0.50 nag·L-1.然而,在小内陆到海边的植被中,土壤湿度、土壤密度、有机质、C含量、总N、pH、有效P、K、Na和Ca含量逐渐降低,而土壤含盐量、Na和Mg含量却逐渐增加.虽然,在植被与相临裸地的不同土壤深度中土壤质地不同,植被地中砂土份额明显(p≤0.01)低于相临裸地,而粉砂土份额则明显(p≤0.001)高于裸地.在本研究中,所有参数的评价也在为其他地区相关研究得到应用.     

Somatic embryogenesis from cell suspension cultures of aspen clone

Suspension cultures initiated from callus derived from petiole explants of aspen hybrid (Populus tremuloides × P. tremula) produced somatic embryos. Callus was induced on a MS medium supplemented with 5 mg·L–1 2,4-D and 0.05 mg·L–1 zeatin under light conditions. Embryogenic calli were obtained when a subsequent subculture of calli was suspended in the same basal me-dium with 10 mg·L–1 2,4-D. The highest number of globular embryos were induced from embryogenic calli by cell suspension cul-ture in a MS liquid medium supplemented with 10 mg·L–1 2,4-D. Genotype and 2,4-D concentration were vital to the induction of embryogenic calli producing competent cells. Embryogenic calli for each genotype were heterogeneous. Green calli with gel-like consistency could yield more competent cells than light yellow embryogenic calli. However, some globular embryos broke into slices and some developed abnormally after one month of culture under the same or other hormonal conditions.     

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.     

Micropropagation of Selected Genotype of Aloe vera L.—An Ancient Plant for Modern Industry

Aloe vera Linn. (Syn. Aloe barbadensis Mill; Gwar-patha in Hindi) belongs to family Liliaceae. The plant, for its medicinal properties, has commercial value. Some of the genotypes of Aloe vera are consumed as a vegetable and processed to make curry and other edible products. We report here on the development of an efficient method for rapid clonal propagation by shoot proliferation from axillary meristem(s) of selected germplasm of Aloe vera. Explants were pretreated with 0.1% aqueous solution of both streptomycin and bavistin separately, each for 15 min. These were surface sterilized with 0.1% aqueous solution of mercuric chloride (HgCl₂) for 4–5 min and washed several times with autoclaved water. These were kept in a chilled, sterile antioxidant (200.0 mg L^?1 of ascorbic acid, 50.0 mg L^?1 of citric acid, and 25.0 mg L^?1 of polyvinylpyrrolidone; PVP) solution and cultured on semi-solid Murashige and Skoog's (MS) medium. The bud explants produced multiple (10.3 ± 0.675/explant) shoots on MS medium containing 13.32 μM of 6-Benzylaminopurine (BAP) and 100.0 mg L^?1 of ascorbic acid, 50.0 mg L^?1 each of citric acid and PVP, with 25.0 mg L^?1 each of arginine and adenine sulphate as additives. The shoots were further multiplied by (a) repeated transfer to fresh MS medium with additives + 13.32 μM BAP, and (b) subculturing on MS medium with a lower (4.44 μM) concentration of BAP. On MS medium containing 4.44 μM of BAP and additives, a maximum number (27.8 ± 0.63) of shoots were produced. In liquid MS medium with 4.44 μM of BAP, the rate of shoot multiplication increased and the vigor of the shoots improved. One hundred percent of the cloned shoots rooted under in vitro conditions on hormone-free half-strength MS salts containing 200.0 mg L^?1 of activated charcoal at 32 ± 2°C. The cloned shoots treated with 2.46 mM of indole-3-butyric acid (IBA) or 2.473 mM of β-naphthoxyacetic acid (NOA) for 5 min rooted under ex vitro conditions in the greenhouse. The rooted plants were hardened in the greenhouse and stored under an agro-net house. The cloned plants were transferred under different field conditions at various sites in Western Rajasthan. These plants grew normally. The higher rate of shoot multiplication and easier approach of direct rooting and hardening make this method superior to the methods previously reported on cloning/tissue culture of Aloe species. From a single shoot bud, approximately 5000 plants can be produced within 180 days.     

Clonal propagation ofGmelina arborea Roxb. byIn vitro culture

In vitro propagation technique ofGmelina arborea multipurpose and a fast growing tree species was studied. Nodal segment including axillary bud was used as a explant. They were cultured on MS media containing various concentrations (0–10 mg/l) of BAP alone or in combination with 0.002 mg/l of IBA. Nodal segments showed axillary bud proliferation in almost all media tested. MS media containing 0.22 mg/l of BAP alone and 2 mg/l of BAP in combination with 0.002 mg/l of IBA were effective for inducing multiple shoots and shoot elongation. MS medium supplemented with 0.02 mg/l of NAA and 1 mg/l of IBA gave the best result for rooting. The regenerated plantlets were potted and acclimatized successfully in a growth chamber and then moved to the green house. Adventitious shoots production from stem explants that were taken from regenerated plantletin vitro was also discussed. Stem segments were tested for their morphogenetic potential on MS media with various combinations and concentrations of BAP, zeatin and TDZ. Successfull result was obtained on MS media supplemented with 2 mg/l of BAP and 1 mg/l of zeatin or supplemented with 0.5 mg/l of BAP and 0.5 mg/l of TDZ. The shoots obtained on MS media containing 2 mg/l of BAP and 1 mg/l of zeatin rooted on MS media containing 0.02 mg/l of NAA and 1 mg/l of IBA, and plantlets were successfully obtained. A part of this paper was presented at the 109th Annual Meeting of the Japanese Forest Society (1998).     

墨兰离体快繁研究

采用墨兰的茎尖和芽为外植体,研究墨兰的组织培养要点。通过对墨兰组织培养中外植体的选择、芽的诱导、继代增殖、壮苗培养、生根诱导和移植进行研究,运用正交试验筛选出芽快速生芽的最佳培养基为:MS+6 BA4.0mg·L-1+NAA1.0mg·L-1+琼脂8g·L-1+蔗糖30g·L-1,生根的最佳培养基为:1/2MS+NAA3.0mg·L-1。分析比较出芽长为1.0cm,叶长为1.5cm,叶数为2～3条的兰花苗有利于生根。