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.     

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.     

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.     

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.     

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.     

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.     

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.     

Dissolved organic carbon and nitrogen in precipitation, throughfall and stemflow fromSchima superba andCunninghamia lanceolata plantations in subtropical China

Despite growing attention to the role of dissolved organic carbon (DOC) and dissolved organic nitrogen (DON) in forest nutrient cycling, their monthly concentration dynamics in forest ecosystems, especially in subtropical forests only were little known. The goal of this study is to measure the concentrations and monthly dynamics of DOC and DON in precipitation, throughfall and stemflow for two plantations ofSchima superba (SS) and Chinese fir (Cunninghamia lanceolata, CF) in Jianou, Fujian, China. Samples of precipitation, throughfall and stemflow were collected on a rain event base from January 2002 to December 2002. Upon collection, all water samples were analyzed for DOC, NO₃ ⁻−N, NH₄ ⁺−N and total dissolved N (TDN). DON was calculated by subtracting NO₃ ⁻−N and NH₄ ⁺−N from TDN. The results showed that the precipitation had a mean DOC concentration of 1.7 mg·L⁻¹ and DON concentration of 0.13 mg·L⁻¹. The mean DOC and DON concentrations in throughfall were 11.2 and 0.24 mg·L⁻¹ in the SS and 10.3 and 0.19 mg·L⁻¹ in the CF respectively. Stemflow DOC and DON concentrations in the CF (19.1 and 0.66 mg·L⁻¹ respectively) were significantly higher than those in the SS (17.6 and 0.48 mg·L⁻¹ respectively). No clear monthly variation in precipitation DOC concentration was found in our study, while DON concentration in precipitation tended to be higher in summer or autumn. The monthly variations of DON concentrations were very similar in throughfall and stemflow at both forests, showing an increase at the beginning of the rainy season in March. In contrast, monthly changes of the DOC concentrations in throughfall of the SS and CF were different to those in stemflow. Throughfall DOC concentrations were higher from February to April, while relatively higher DOC concentrations in stemflow were found during September–November period. Foundation item: This study was supported by the Teaching and Research Award program for MOE P.R.C. (TRAPOYT). Biography: Guo Jian-fen (1977-), female, Ph. Doctor in College of Life Science, Xiamen University, Xiamen 361005, P.R. China. Responsible editor: Zhu Hong     

Establishment of a transgenic system in fast-growing black locust （Robinia pseudoacacia L.）

The AhDREB1 gene, cloned from Atriplex hortensis L., was transferred into black locust （Robiniapseudoacacia L.） by an Agrobacterium-mediated transformation. The results suggest that stems of black locust sub-cultured in vitro for 20 d are suitable for genetic transformation. The optimum concentrations of kanamycin and cefotaxime were 30 and 150 mg.L-1, respectively. Important factors affecting the transformation efficiency were studied by means of a L9（3^4） orthogonal design. An effective system for genetic transformation in black locust was developed as follows： the stems were pre-cultured for 2 d, immersed in the Agrobacterium solution （OD600 = 0.7） with 10 mg·L^-1 acetosyringone for 21 min and then co-cultured for 2 d. The selection pressures, changing from low to high, could improve transformation efficiency. The transgenic plants were identified by a PCR method. The PCR results indicated that the AhDREB1 gene had been integrated into the genome of black locust and two lines of the transgenic plants were obtained.     

Effects of elevated CO2 concentrations on photosynthesis, dark respiration and RuBPcase activity of three species seedlings in Changbai Mountain

Two-year-old seedlings ofPinus koraiensis, Pinus sylvestriformis andFraxinus mandshurica were treated in open-top chambers with elevated CO₂ concentrations (700 μL·L⁻¹, 500 μL·L⁻¹) and ambient CO₂ concentrations (350 μL·L⁻¹) in Changbai Mountain from June to Sept. in 1999 and 2001. The net photosynthetic rate, dark respiration rate, ribulose-1,5-bisphosphate carboxlase (RuBPcase) activity, and chlorophyll content were analyzed. The results indicated the RuBPcase activity of the three species seedlings increased at elevated CO₂ concentrations. The elevated CO₂ concentrations stimulated the net photosynthetic rates of three tree species exceptP. sylvestriformis grown under 500 μL·L⁻¹ CO₂ concentration. The dark respiration rates ofP. koraiensis andP. sylvestriformis increased under concentration of 700 μL·L⁻¹ CO₂, out that ofF. mandshurica decreased under both concentrations 700 μL·L⁻¹ and 500 μL·L⁻¹ CO₂. The seedlings ofF. mandshurica decreased in chlorophyll contents at elevated CO₂ concentrations. Foundation item: This paper was supported by the National Natural Science Foundation of China (No. 30070158). Knowledge Innovation Item of Chinese Academy of Sciences (KZCX2-406) and “Hundred Scientists” Project of Chinese Academy of Sciences. Biography: Zhou Yu-mei (1973-) Ph. Doctor, Assistant Research fellow Institute of Applied Ecology. Chinese Academy of Sciences. Shenyang 110016. P.R. China. Responsible editor: Song Funan     

Distribution of gymnemic acid in various organs of Gymnema sylvestre

The gymnemic acid contents in various organs of Gymnema sylvestre were investigated by High Performance Liquid Chromatography (HPLC) method. The results shows that the content of gymnemic acid in various organs, obviously different, was 54.29, 31.66, 28.82, 27.67, 25.39, 20.56 and 1.31 mg·g⁻¹ DW in shoot tips, flowers, nodes, leaves, internodes, roots and seeds, respectively. The highest gymnemic acid content (54.29 mg·g⁻¹ DW) was found in shoot tip, 1.96 fold higher than that in leaves (27.67 mg·g⁻¹ DW). Maximum quantity of gymnemic acid (35.39 mg·g⁻¹ DW) was observed in the young leaves, which was 1.52 times higher than that in old leaves (23.07 mg·g⁻¹ DW). The content of gymnemic acid in young, middle and old internodes was 26.47, 25.77 and 23.94 mg·g⁻¹ DW, respectively, all lower than that in leaves (27.67 mg·g⁻¹ DW), whereas the content of gymnemic acid in young, middle and old nodes was 27.96, 28.81 and 29.66 mg·g⁻¹ DW, respectively, all higher than that in leaves. The study provides the scientific evidences for the rational development and utilization of Gymnema sylvestre resources, since over exploitation of natural stands has caused depletion of these plants in nature. Foundation project: This work was supported by University Grants Commission, New Delhi (No. 33-88/2007 (SR))     

Response of seedlings growth ofPinus sylvestriformis to atmospheric CO2 enrichment in Changbai Mountain

The biomass and ratio of root-shoot ofPinus sylvestriformis seedlings at CO₂ concentration of 700 μL·L⁻¹ and 500 μL·L⁻¹ were measured using open-top chambers (OTCs) in Changbai Mountain during Jun. to Oct. in 1999. The results showed that doubling CO₂ concentration was benefit to seedling growth of the species (500 μL·L⁻¹ was better than 700 μL·L⁻¹) and the biomass production was increased in both above-ground and underground parts of seedlings. Carbon transformation to roots was evident as rising of CO₂ concentration. This project is supported by Chinese Academy of Sciences Responsible editor: Chai Ruihai     

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.     

Effect of natural brassinolide on germination of Ailanthus altissima seeds

The germination capacity of Ailanthus altissima seeds improved after the seeds were soaked with different concentrations of natural brassinolide (NBR). The germination rate and germination energy of the seeds increased by 17.6% and 18.8%, and the mean germination speed (i.e., germination time) of the seeds was shortened by 1.4 d under the optimal concentration (0.4 mg.L⁻¹) treatment, compared with the control. After hypocotyls of A. altissima were treated with NBR, the elongation of the hypocotyls increased. Among different concentrations of the NBR, 0.4 mg·L⁻¹ NBR appeared to be the optimal concentration for the elongation of A. altissima hypocotyls.     

外源糖溶液和高浓度CO2处理对白桦叶片糖和蛋白质含量的影响

3年生白桦同时接受3种外源糖溶液(蔗糖、果糖、葡萄糖)和3种高浓度CO2(700、1400、2100μL·μL-1CO2)处理.处理1个月后,测定了叶片的总糖、蔗糖、果糖和蛋白质含量.结果表明:在700μL·L-1和1400μL·L-1 CO2下,外源糖溶液增加了叶片的可溶性糖和蛋白质含量,其中外源蔗糖的效果最好:外源糖溶液与2100μL·L-1CO2结合,会抑制叶片积累总糖和蛋白质:在700μL·L-1和1400μL·L-1CO2下,喷施葡萄糖、果糖的叶片在蛋白质含量上没有明显差别:同700、1400μL·L-1CO2相比,除喷施果糖植株外,2100μL·L-1 CO2明显增加了叶片的总糖、蔗糖、果糖和蛋白质含量:在喷施同种外源糖溶液的情况下,叶片的糖含量与CO2浓度呈正相关性.图6参7.     

Adventitious shoot regeneration of Platanus acerifolia Willd. facilitated by Timentin, an antibiotic for suppression of Agrobacterium tumefaciens in genetic transformation

The effects of Timentin and cefotaxime (Cef) on shoot regeneration of the London plane tree (Platanus acerifolia Willd.) and their use for the suppression of Agrobacterium tumefaciens in Agrobacterium-mediated genetic transformation were compared. Shoot regeneration was significantly reduced on the media with Cef at concentrations from 100 to 500 mg·L–1. Timentin showed negative effect on plant regeneration at concentrations of 100 and 500 mg·L–1; however, 300 mg·L–1 Timentin was shown to facilitate shoot r...     

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     

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     

Photosynthetic response ofPinus sylvestriformis to elevated carbon dioxide and its influential factor analysis

The photosynthetic response of 12-year oldPinus sylvestriformis to elevated CO₂ and its influential factors were tested and analyzed in the forest region of Changbai Mountain in 1999. Trees grown at the natural condition were controlled at three levels of CO₂ concentration (350 μL·L⁻¹, 500 μL·L⁻¹ and 700 μL·L⁻¹) by CO₂ rich settlement designed by us. Net photosynthetic rates (NPR), temperature, relative humidity, stomatal conductance, intercellular CO₂ concentration and photosynthetic active radiation (PAR) were measured at 6:00, 8:00, 10:00, 14:00, 16:00 and 18:00 hours a day. Experimental results showed that the NPR ofPinus sylvestriformis increased by 32.6% and 123.0% at 500 μL·L⁻¹ and 700 μL·L⁻¹ CO₂ concentration respectively, compared to ambient atmospheric CO₂ concentration (350 μL·L⁻¹). The relations between NPR and influential factors, including temperature, relative humidity, intercellular CO₂ concentration and photosynthetic active radiation, were analyzed respectively by regression analysis at different CO₂ concentrations. Foundation Item: This project was supported by Chinese Academy of Sciences. Biography: WANG Chen-rui (1970-), male, Assistant Research Fellow in Institute of Applied Ecology, Chinese Academy of Sciences. Responsible editor: Chai Ruihai