Establishment of in vitro culture of Populus euphratica Olivier

The purpose of our study was to establish a regeneration system for micropropagation of Populus euphratica Olivier. On the basis of an analysis of plant leaf mineral nutrients, a special medium was proposed, called MP2. In optimizing media for in vitro plant cultures including MS, B5 and MP2 media we employed hormones, auxin IAA, cytokine benzyladenine (BAP) and gibberellic acid (GA) in our factorial experiments on media. Adventitious shoots were derived from cuttings of adult plants taken from Xingjiang, west China, on selected media with MP2 0.5 mg·L-1 BA 0.1 mg·L-1 NAA. The shoots were elongated on a medium with 0.25 mg·L-1 BAP, 0.1 mg·L-1 NAA and 2 mg·L-1 GA and were then rooted on a medium with 0.2-0.5 mg·L-1 IBA. All the media were incorporated with 30 g·L-1 sucrose and an adjusted pH at 6.3.     

Inhibitive effects of some treatments on the browning rate during the in vitro culture of Acacia karroo Hayne

Acacia karroo Hayne is an arbor species widely distributed in South Africa with the characteristics of fast growth and drought resistance. The species was introduced to China recently. In vitro culture is an effective method to rapidly produce plants and a strategy to minimize somaclonal variation among regenerated plants. Browning, however, is a problem in establishing the in vitro culture system. The present study diminished the problem by selecting explants, using different browning inhibitors and chilling treatment. Results showed that the use of embryos as explants reduced the browning ratio to 46.7%, whilst stem segment explants were browned up to 56.7%. The adventitious buds were successfully induced in the modified tissue culture medium supplemented with 5.0 mg·L-1 6-BA and 0.1 mg·L-1 NAA. The proliferation coefficient of adventitious buds is 2.8.     

Cloning of plasma membrane H-ATPase gene in Populus euphratica Oliv.

For this paper, the plasma membrane (PM) H -ATPase gene has been cloned from Populus euphratica Oliv. through a homology based strategy. The isolated 3,210 bp cDNA contains a single 2,862 bp open reading frame (ORF) which encodes a putative H -ATPase protein of 953 amino acid residues, with a significant homology to plasma membrane H -ATPase of Primus persica, Phaseolus vulgaris, Sesbania rostrata and Daucus carota. The predicted protein has a molecular weight of 104,553 Da. The copy number analysis revealed multiple copies of the PM H -ATPase in the P. euphratica genome after digestion of their genomic DNA by the restriction enzymes EcoRI, NdeⅠ, FbaⅠand BglⅡ, and Southern blot.     

Agrobacterium tumefaciens-mediated GUS gene transformation of Robinia pseudoacacia ‘Idaho’

Based on the plant regeneration system, a GUS gene transformation system to Idaho locust (Robinia pseudoacacia ‘Idaho’) mediated by Agrobacterium tumefaciens was established. The successful transformation was confirmed by regenerating the shoots from the infected leaves in the presence of hygromysin; by histochemical X-gluc assays of β-glucuronidase (GUS) and by PCR and PCR-Southern blotting analysis. The ratio of positive transgenic plants is 5.8% (5 out of 86 plants). With this system, the target gene DREB was introduced into the leaves of Idaho locust. The transgenic plants regenerated, which was verified by PCR-Southern blot-ting. It is suggested that the transformation system could be a new, simple, reliable and practical route to gene transformation of R. pseudoacacia ‘Idaho’ mediated with A. tumefaciens.     

Construction and Characterization of cDNA Library from Water-Stressed Plantlets Regenerated in vitro of Populus hopeiensis

In order to isolate and clone water-stress-responsive genes, total RNA was extracted from water-stressed plantlets regenerated in vitro of Populus hopeiensis using a QIAGEN RNeasy Plant Mini Kit. CDNA, synthesized by LD-PCR with the SMART cDNA Library Construction Kit, was n vitro packaged into a phage λTriplEx2 vector. The resulting primary library and amplified library have a titer of 1.68×10i6 and 1.69×109 pfu·mL–1 respectively. The combination ratio reached 98.8% and the average size of inserts was about 800 bp. In addition, the percentage of inserted fragments (> 400 bp) was approximately 90%. The results indicate that a cDNA library has been successfully constructed.     

Rooting and acclimatization of in vitro plantlets of mtl-D gene modified Robinia pseudoacacia ‘Idaho’

Robinia pseudoacacia 'Idaho' (Robinia×ambigua 'Idahoensis', R. pseudoacacia×R. viscosa) modified by a mtl-D gene went through five lines and had characteristics of drought tolerance. Three stages of their micropropagation had been studied by previous investigators. The other two stages, in vitro shoot rooting and plantlet acclimatization, still remained unsolved in the laboratory. For this paper, we studied the later two stages based on the previous achievements. Results showed that the highest rooting rate of Idaho locust was 98.4% when the in vitro shoots, over 2.5 cm in height and 0.08 cm in diameter, were placed on a half strength MS basal medium with 0.4 mg·L-1 IB A and 0.1 mg·L-1 NAA as supplements and were solidified with 0.5% agar; the highest survival rate was 98.3% when the rooted plantlets were potted in vermiculite. All the stages for micropropagation of the Idaho locust, modified by the mtl-D gene, were assembled completely. The tissue culture plants grow well in the field.     

High-level expression of 4-coumarate:coenzyme A ligase gene Pt4CL1 of Populus tomentosa in E. coli

In order to investigate the enzymatic properties of the 4CL1 of Populus tomentosa, the recombinant expression vector pQE31-4CL1 was constructed. The recombinant was identified by three restriction endonucleases, then the vector pQE31-4CL1 was transformed into expression host M15 (pREP4) and induced by isopropyl-α-D-thiogalactoside (IPTG) to express 60 kD fused protein Pt4CL1. The biologically active Pt4CL1, expressed as soluble protein, was achieved with 0.6 mmol·L-1 IPTG induction as the ex-pression temperature declined from 37 to 28°C. The 6×His tag facilitates affinity binding to Ni2 -nitrolotriacetic acid (NTA) and enables one-step purification to acquire the molecular SDS-PAGE electrophoresis purity of the active 4CL1 protein by agarose cou-pled with Ni2 -NTA affinity chromatography. The optimal substrate for Pt4CL1 was 4-coumarate.     

Molecular Cloning of a Cellulose Synthase Gene PtoCesA1 from Populus tomentosa and Its Genetic Transformation in Tobacco

A 3 125 bp cellulose synthase gene, PtoCesA1, which has a 98% identity to PtrCesA1 from Populus tremuloides, was cloned from cDNA prepared from secondary xylem of P tomentosa. Four anti-expression vectors with different fragments of PtoCesAl, named as pBIPF, pBICC1, pBIPR and pBIBR, were constructed. Some traits of transformed tobacco of pBICC1, pBIPR and pBIBR differed from wild types, such as small leaves, ＂dwarf＂ phenotype and thinner xylem and fiber cell walls than wild plants consistent with a loss of cellulose. It indicated that the growth of transgenic tobacco was restrained by the expression of anti-PtoCesA1. Transgenic tobacco was obtained and the contents of cellulose and lignin were analyzed as well as the width and length of fiber cells, and xylem thickness for both transgenic and control plants. Transformed tobacco showed a different phenotype from control plants and it implied that PtoCesA1 was essential for the cellulose biosynthesis in poplar stems.     

Liquefaction and FTIR analysis of two desert shrubs Salix psammophila and Caragana intermedia

1 Introduction Desert shrubs are made up of natural composite mate- rials, composed of cellulose, hemicellulose and lignin. Although there are a large number of hydroxyl groups in them, these are not fully used due to the high crys- tallinity of cellulose and tight interconnection among polymeric components in wood (Cheng et al., 2002). Liquefaction of wood is a new technique in wood in- dustries. It is a novel process which effectively con- verts the chemical composition of wood into macro- …     

Natural Resistance of Two Plantation Woods Populus × canadensis cv. and Cunninghamia lanceolata to Decay Fungi and Termites

Natural durability of two plantation woods, Chinese fir and I-214 poplar, was investigated thoroughly by three testing methods, namely an accelerated laboratory decay test, a fungus cellar test and a field test. After the decay test using Postia placenta and Trametes versicolor, Chinese fir and the I-214 poplar showed 34% and 69% of mass loss, respectively, indicating they should be classified as slightly durable and non-durable wood. This conclusion was confirmed by the fungus cellar test and the field test. Like the performance in the decay test, I-214 poplar showed no resistance to termites either in the laboratory or in the field, whereas Chinese fir would be classified as moderately resistant. [Supported by Sino-Japanese Technical Cooperation Project Titled “Studies on Chinese Plantation Wood” (JICA PROJECT/033-1418-E-O)]     

Optimization of the uidA Gene Transfer of Rosa hybrida via Agrobacterium tumefaciens: an Assessment of Factors Influencing the Efficiency of Gene Transfer

To develop a transformation protocol of Rosa hybrida ‘Samantha‘ via Agrobacterium tumefaciens, the authors examined the effect of different factors on T-DNA transfer by measuring transient expression levels of an intron-containing β-glucuronidase gene. The results indicate that explant, light condition, salt concentration and acetosyringone (AS) concentration in co-culture medium are the most important factors, and factors like co-culture temperature, co-culture period and bacteria density have a strong effect on the growth of bacteria and then T-DNA transfer. Optimized co-cultivation was performed by inoculation of embryogenic callus with bacteria at a density of OD600=0.5-0.8 for 20 min and co-culture in darkness under 23 ℃ on medium with 1/2 MS salts and 300 μmol-L 1 AS for 3 d.     

Effects of Bamboo Fargesia murielae on Plant Diversity in Fir Forest on Mountain Shennongjia

Simultaneous flowering caused a gregarious dieback of umbrella bamboo (Fargesia murielae) all over the world in 1993-2000. Mountain Shennongjia in Central China‘s Hubei Province is the only native home of umbrella bamboo, where it dominates the understorey of the farges fir (Abiesfargesii) forest between 2 400 and 3 100 m, covering the ground for more than one century before a periodically flowering death. Data from 20 quadrats along a vegetation sequence revealed that the density, coverage,and height of umbrella bamboo negatively affected the species richness, diversity, and evenness of vascular plants in the forest. Local climax community of fir-bamboo was the poorest in species diversity, while the bamboo-free communities (i.e. shrubs and meadows)were bearing a higher species diversity. The simultaneous dieback of umbrella bamboo is an ecological relcase, which can pcriodically promote the plant diversity in the fir forest.     

Regeneration and gene transformation systems of Robinia pseudoacacia ‘Idaho’ mediated by Agrobacterium tumefaciens

Robinia pseudoacacia ‘Idaho’ is one of several multi-purpose trees used in ornamental, soil and water conservation, fodder and nectar sources. Plant abiotic stress tolerance transformed by genes could meet the requirements for reclamation of arid or alkalid lands and vegetation restoration. For this paper, we studied the effects of auxin and cytokine on Idaho locust in vitro regeneration and the establishment of gene transformation systems for plants mediated by Agrobacterium tumefaciens. Results showed that the ratios of cytokinin and auxin were the major factors affecting adventitious bud differentiation on a MS medium; the concentration of 0.5 mg•L^–1 6-BA benefitted callus proliferation and 0.25 mg•L^–1 IBA promoted shoot rooting; however, a higher IBA concentration will inhibit rooting. The most effective antitoxin for screening transgenic Idaho locust shoots was G418 and the most sensitive concentra-tion of it was 8 mg•L^–1.     

Lignin Characterization of Triploid Clones of Populus tomentosa Carr.

In order to understand the structural characteristics of lignin in triploid clones ofPopulus tomentosa and its changes in the processes of pulping and bleaching, milled wood lignin （MWL）, lignin carbohydrate complex （LCC） and the residual lignin from kraft pulp （KP） and sulfite pulp （SP） were isolated and analyzed by Fourier transform infrared （FTIR） spectrum and ^13C nuclear magnetic resonance （NMR）. The most diagnostic peaks were assigned and the differences were discussed. The spectral patterns reveal that triploid P tomentosa shows the specific features of hardwood from temperate areas, but in the spectrum of FTIR, the strength ratio orAl270 cm^-1 to A1226 cm^-1 is 0.88, higher than the average of hardwood from temperate areas, which will make the lignin delignification more difficult during pulping and bleaching. The LCC from triploid P tomentosa is mainly composed of xyloglucan and glucuronic acid, and other glucides have much lower ratio. In LCC FTIR, there are three peaks at 1 427, 1 329 and 1 046 cm^-1, indicating that both semi-cellulose and cellulose could exist in LCC, and that there might be relationships between cellulose and lignin. Compared with the residual lignin from KP and SP, the condensed structure in KP is more than that in SP.     

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 hypocoryls 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.     

Pistil Development in 2 Types of Flowers of Xanthoceras sorbifolia

In order to investigate flower development and female abortion during sex differentiation ofXanthoceras sorbifolia,anatomical observations and comparative study on differential proteins were carried out in different developmental stages of two types of flowers of this species. It was found that the selective abortion happened in male flower before the formation of megasporocyte. Special proteins related to the female abortion were found through 2-dimensional electrophoretic analysis. Protein Al(14.2 kD)only existed in florescence of male flower, while B1 (13.7 kD) and B2 (18.2 kD) disappeared in that stage of male flower. They were all considered to be relative to pistil abortion ofXanthoceras sorbifolia.     

Microwave vacuum drying characteristics of Pinus massoniana wood

Microwave-vacuum (MV) drying characteristics of plantation Masson pine (Pinus massoniana) were studied experimen-tally for various levels of microwave radiation time, initial moisture content (MC), vacuum level and wood thickness. The results show that the process of MV drying for wood can be significantly divided into a short accelerating rate drying period, a long constant rate drying period and falling rate drying period, and the second drying period can extend to levels of mean MC below the fiber satu-r...     

Research advances in allelopathy of Quercus L.

The term allelopathy refers to biochemical interaction between all types of plants including microorganisms through pro-duction of chemical compounds that escape into the environment and exist widely in Quercus L. The development of investigations into types of allelochemicals, their compounding and spreading pathways, as well as expression of allelopathy in Quercus L. are re-viewed in this paper. We have closely and systematically emphasized the functional mechanisms of allelopathy in forest plants for our next research project on allelopathy in Quercus L.     

Twelve Elastic Constants of Betula platyphylla Suk.

Wood elastic constants are needed to describe the elastic behaviors of wood and be taken as an important design parameter for wood-based composite materials and structural materials. This paper clarified the relationships between compliance coefficients and engineering elastic constants combined with orthotropic properties of wood, and twelve elastic constants of Betula platyphylla Suk. were measured by electrical strain gauges. Spreading the adhesive quantity cannot be excessive or too little when the strain flakes were glued. If excessive, the glue layer was too thick which would influence the strain flakes?performance, and if too little, glues plastered were not firm, which could not accurately transmit the strain. Wood as an orthotropic material, its modulus of elasticity and poissons ratios are related by two formulas:μij /Ei =μji /Ej and μij < (Ei /Ej)1/2. The results showed that the elastic constants of Betula platyphylla Suk. measured by electrical strain gauges were accurate and reliable. The results of shear elastic modulus GTL and GLR show a high linear regression correlation coefficient (> 0.95) between the reciprocal of elastic modulus MOE-1 and the square of the ratio of depth to length (h/l)2, which indicate that shear modulus values measured were reliable by three point bending experiment.     

Polyploid induction of Lespedeza formosa by colchicine treatment

Polyploid induction has been conducted by different colchicines concentrations with seed, hypocotyl and apices of Lespe-deza formosa. Morphological variation and amounts of chromosome about the polyploidy have also been analyzed. Results show that the best material for polyploid induction of L. formosa is the new apices from seed germination, with an induction rate of 44.4% at 0.1% colchicines concentration and 36 hours of treatment time. Compared with normal diploid plants, the polyploidy plants of L. for-mosa inducted in our experiments have short stature and stems with thick and wide leaves. Cytological studies show that the changed seedlings, whose morphology has changed dramatically, are tetraploid, with 44 (2n = 4x = 44) chromosomes in the somatic cells.