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- …     

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.     

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.     

Cloning and RNAi Construction of a LEAFY Homologous Gene from Populus tomentosa and Preliminary Study in Tobacco

PtLFY, a LEAFY （LFY） gene, was cloned from Populus tomentosa （LM50） by PCR. Sequencing analysis indicated that PtLFY was 2629 bp long, composed of three exons and two introns and encoded 378 amino acids. The splice donor sites and the splice acceptor sites were in identical positions to the LFY and its homologues. The amino acid sequence inferred was 68%-99% homologous to those of LFY and its homologues by blast analysis in GenBank. The Southern blot analysis indicated that there was a single copy of the PtLFY gene in genomic DNA of male and female P. tomentosa （LM50 and 5082）. The pBI121-Ptalfy （reverse）-intron-Ptlfy-GUS-nos was constructed using RNA interference （RNAi） technique and verified by PCR and digestion identification and transformed into tobacco. Some transgenic tobacco plants were obtained by PCR and PCR-Southern identification. The growth was generally repressed in transgenic tobacco plants compared with wild-type ones and some phenotypic differences were observed.     

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.     

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.     

Agrobacterium tumefaciens-mediated GUS gene transfer to Sophora japonica L.

Agrobacterium-mediated genetic transformation of Sophora japonica was standardized using the Agrobacterium tumefaciens strain LBA4404 that harbored the binary vector pBI121 containing genes forβ-glucuronidase (GUS) and neomycin phos-photransterase (nptⅡ). S. japonica transformants were selected by the ability of the leaf explants to produce kanamycin-resistant calli that regenerated into kanamycin-resistant plantlets. Successful transformation was confirmed by histochemical assay for GUS activity, PCR analysis and Southern blot. The period of nearly two months was required for the regeneration of transgenic plantlets from the explants. The transformed plants resembled their parents in morphology.     

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.     

Cross Breeding of Populus and Its Hybrids for Cold Resistance

Populus tomento a was crossed with P. tremuloidis, P grandiden ata, P. lba × P. grandidentata and P. alba × Ulmus s . ta pumila in order to maintain its rapid growth and high wood quality and improve its resistance to cold. Two methods were used to increase the germination rate from 1.5% to 41.1% and the remaining rate from 1.7% to 44.2%. Forty crossing combinations were conducted and 2 744 hybrid seedlings were obtained. MX. t4 × P grandiden ata (G-1-58), MX3 × P. tremuloidis (T-44-60), MX2 × P. tremuloidis (l-13-87-37) and MX2 × (P. alba × P. grandidentata) were regarded as superior combinations after analysis and selection. Thirty seedlings of these combinations and 11 triploid seedlings identified by counting their chromosomes were selected as super plants.     

Floral display and breeding system of Jatropha curcas L.

Plant flowering and breeding characteristics are important for us to understand the reproduction of plant populations. In this paper, we studied the reproduction characteristics of Jatropha curcas in Yuanjiang County (23°36′N, 101°00′E), Yunnan Province. The plant produces flowers in dichasial inflorescences. Normally, the flowers are unisexual, and male and female flowers are produced in the same inflorescence. Only a few male flowers are produced in an inflorescence, and fruits are produced only through pollination between different flowers from the same or different plants. By the treatments of emasculation, bagging and artificial pollination in this experiment, there were few but same fruit set ratios when the inflorescences were emasculated, bagged, or bagged with net, except artificial pollination treatments, which showed that Jatropha curcas could produce fruit through apomixis but not wind pollination. When the inflorescences were unbagged, unemasculated and with free pollination treatments, or bagged, emascu- lated and with artificial cross-pollination treatments, or unbagged, emasculated and with free pollination treatments, there were many fruits produced. It showed that Jatropha curcas shows outcrossing, is self-compatible, and demanding for pollinators. Normally, the male flowers open first and a few flowers bloom in one day in a raceme. These flowers last a long time in bloom. However, a large number of female flowers open from the third to the fifth day, with some female flowers opening first in a few raceme. This shows a tendency to promote xenogamy and minimize geitonogamy.     

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.     

Cold tolerance of Acacia karroo and A. nilotica, two tree species from South Africa, in Beijing

Fynbos plant communities developed under geographic and climatic conditions unique to South Africa. Acacia karroo is the most widespread tree species, well known for its drought tolerance. A. nilotica exudes a good quality gum which is edible and suitable for confectionery. The two species have been introduced to China but little is known about their adaptability under the new environmental conditions. For our present study, the growth potential and physiological and biochemical parameters of the two species in response to cold stress were investigated. The results, measured as seed qualities (purity, thousand seed weights, germination, etc.) show differences between A. karroo and A. nilotica. The seed germination rate was between 10% and 60%. Seedlings were established in fields, pots and plugs. Those in the field grew well during the growing season but completely died in the winter. Those in pots or plugs were moved to and managed in the greenhouse. The plants were green in winter. The electrical conductivity of cells, the contents of water soluble sugars, proteins, malondialdehyde (MDA) increased during eight hours of low temperature treatment at 4℃. The activities of superoxide dismutase (SOD), peroxidase (POD) and catalase (CAT) decreased markedly during the eight hours of 4℃treatment. We conclude that A. karroo and A. nilotica have little adaptability to the cold winter weather and cannot survive in the outdoors in the Beijing area, but could grow well indoors as ornamentals.     

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.     

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.     

High-efficiency somatic embryogenesis and morphohistology and histochemistry of somatic embryo development in Larix leptolepis Gordon

A high-efficiency somatic embryogenesis protocol of Japanese larch (Larix leptolepis Gordon) has been established in our investigation. Calli were induced from immature zygotic embryos of female cones of L. leptolepis and then subcultured regularly on to a modified Gupta and Durzan (DCR) basal medium for 5 years. Embryogenic tissues showed distinct morphological changes dur-ing somatic embryo development when they were transferred to a maturation medium supplemented with abscisic acid (ABA) com-pared with the morphology in a medium lacking ABA. Histological observations indicated that polyembryony was a characteristic feature during early embryogeny and somatic embryos at later stages showed normal histodifferentiation. In addition, histochemical analysis revealed that abundant starch granules and proteins accumulated in mature embryos, indicating that they played important roles in the development and regeneration of normal plantlets from somatic embryos on hormone-free germination media     

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.     

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.     

Variation Analysis of Seed and Seedling Traits of Cross Combination Progenies in Populus

Twenty-five species and hybrids in Populus were used as parents, and 26 cross combinations, including more than 5 000 seedlings, were obtained by artificial cross breeding. The length of infructescence, number of seeds per infructescence, thou-sand-seed weight, germination rate of seeds among these cross combinations were tested. The results indicated that the cross combi-national effects were significant for these traits, and demonstrated that the length of infructescence, thousand-seed weight were posi-tively affected by female parent. In addition, seedling height, diameter above ground, diameter at breast height (DBH) of 17 cross combination progenies were investigated. The analysis of mean and standard deviation of these three traits showed that seedling height, diameter above ground, DBH had extensive variation among combinations and individuals within combination. Variance analysis and estimate of heritability indicated that the three traits had wide variation and were controlled by heredity. It was feasible to select superior cross combinations and seedlings. Further more, the result of multiple comparison showed that P. deltoides ‘Lux’ × P. deltoides ‘D324’, P ussuriensis cl. ‘U4’ × P. deltoides ‘T66’, P. ussuriensis cl. ‘U4’ × P deltoides ‘T26’, P. deltoides ‘Lux’ × P.. . ussuriensis cl. ‘U3’, (P. m tos × P. bolleana) × (P alba × P. glandulosa), (P. alba × P. tomentosa) × (P. al × P. glandulosa ), and to en a . ba (P. alba × P. glandulosa ‘No. 2’) × P. tomentosa ‘Lumao 50’ were superior cross combinations with higher growth rate. Finally, 123 elite seedlings were selected for further test.