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...     

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.     

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 different treatments on seed germination and breaking seed dormancy of <Emphasis Type="Italic">Prosopis koelziana</Emphasis> and <Emphasis Type="Italic">Prosopis Juliflora</Emphasis>

For improving seed germination of Prosopis koelziana and Prosopis juliflora, different treatments of seeds were conducted, including scarification with sulfuric acid 98% for 10 and 15 min, sandy paper, hot water for 5 and 10 min, potasium nitrate 0.1%, gibberellic acid at 250 mg·L⁻¹ and 500 mg·L⁻¹ and combinational treatment of scarification with gibberellic acid of 250 mg·L⁻¹ and 500 mg·L⁻¹. The results show that scarifications with sandy paper and sulfuric acids 98% were the most effective treatments on breaking seed dormancy and seed germination induction. Scarification with sulfuric acid 98% for 15 min was the best treatment. According to the positive effect of scarification and lack of reaction of seeds against KNO₃ and gibberellic acid, the kind of seed dormancy was determined as exogenous.     

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.     

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 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.     

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.     

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.     

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.     

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.     

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.     

Contamination and browning in tissue culture of Platanus occidentalis L.

Twigs of 2–3-year-old Platanus occidentalis L. were used as experimental material to find the causes for the contamination and browning in the initial stages of tissue cultures. To compare the degree of browning of explants picked off from different growing seasons, the experimental material was excised from trees on each of the first ten days in January, March, May and July, 2006. The results indicated that the contamination and browning rates of the material cut off in January (14.2% and 30.6%, respectively) and March were somewhat lower than those in July. The pretreatment of soaking the explants in different anti-oxidants and absorbents at the same time could diminish some side effects. The pretreatment of using 10 g·L–1 vitamin C reduced the contamination and brown-ing rate effectively. An orthogonal experiment showed that the optimal factor and level arrangement is 0.5 mg·L–1 BA, 2.0 g·L–1 ac-tive carbon and 1.5 g·L–1 PVP which resulted in a browning rate of only 16.5%. In general, sampling period, physical properties and pretreatment of explants are the main factors responsible for the contamination and browning of material in the initial stages of P. occidentalis tissue cultures.     

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.     

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     

冷湿层积法对印度冷杉(Abies pindrow)和长叶云杉(Picea smithiana）种子萌发的影响

研究赤霉素(GA3),冷湿和温度 对五个种源的印度冷杉(Abies pindrow)和长叶云杉(Picea smithiana)种子萌发的影响.种子被浸泡在GA3 (10 mg(L-1)中24小时,然后在3(5(C温度的条件下冷藏15天.设计4个温度(10(C, 15(C, 20(C 和 25(C)条件来促进种子的萌发.结果表明,浸泡和冷湿处理明显增加种子的萌发率.在10 (C时种子的萌发率最高.总体结果表明,种子被浸泡在GA3 (10 mg(L-1)中24小时,冷湿藏15天,可以有效地促进印度冷杉和长叶云杉的种子萌发.统计数据表明,浸泡处理、温度和种源以及与温度的相互作用都对种子的萌发有明显的影响.     

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     

Effect of NaCl on growth and ion relations in two salt-tolerant strains of Paxillus involutus

The effect of NaCl on growth, biomass and ion relations of two salt-tolerant isolates of Paxillus involutus, MAJ and NAU were investigated. The two Paxillus strains were exposed to the following concentrations of NaCl： 0, 100, 200 and 500 mmol·L^-1. Growth of MAJ and NAU was enhanced by 100 mmol·L^-1 NaCl but severely inhibited at the concentration of 500 mmol·L^-1. NAU exhibited a greater capacity to exclude Na^＋ and Cl^- under all salinity levels, whereas the salt-includer MAJ had a higher capacity in nutrient uptake under salt stress. The ratios Na^＋/K^＋, Na^＋/Ca^2＋ and Na^＋/Mg^2＋ in NaCl-treated MAJ and NAU did not increase at levels of 100 and 200 mmol·L^-1 NaCl but markedly increased at 500 mmol·L^-1. This suggests that the two strains, especially MAJ, enhanced nutrient uptake corresponding to the increased Na^＋ influx at moderate salinity. We conclude that both MAJ and NAU are able to tolerate 500 mmol·L^-1 NaCl but there are species-specific differences in retaining ionic homeostasis in the two Paxillus strains. NAU is a salt-excluder, MAJ is a salt-includer but retains a high capacity in nutrient selectivity under saline conditions. Their definite mechanisms to enhance salt tolerance of mycorrhizal hosts need further study.     

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))