转多基因库安托杨非靶目标材性性状分析

In this study, the transgenic poplar (Populus × euramericana‘Guariento’) harboring five exogenous genes (SacB, Vgb, BtCry3A, OC-I and JERF36^#) and the control plants were selected, comparing the wood property variation of 5-year-old trees, and trying to understand whether the transgenic of exogenous genes produce unintended effects. The results showed that the transgenic of exogenous genes caused a wide range of variation in non-target wood properties, the variations were found not only between transgenic and wild-type lines, but also among transgenic lines. Compared with the control, the transgenic lines, the contents of holocellulose and cellulose increased, the total lignin and acid soluble lignin decreased, the fiber length, the fiber width, the microfibril angle and the acid insoluble lignin varied uncertainly. It is concluded that the introduced exogenous gene could generate unintended effects on corresponding non-target target traits.     

Determination of drought tolerance using root activities in Robinia pseudoacacia ‘Idaho’ transformed with mtl-D gene

Idaho locust (Robinia pseudoacacia ‘Idaho’) is an exotic multi-purpose tree used in landscaping, soil and water conservation, fodder sources and others. To improve its drought tolerance for reclaiming arid land, five lines of transformed mtl-D gene, as osmotic regulator in plant cells, have been selected and managed to determine their drought tolerance under experimental conditions. Qualitative and quantitative variables of transformed plants were studied. The critical value of drought tolerance was determined by detecting the 2,3,5-triphenyl tetrazolium chloride (TTC) reductants in roots and soil water content (SWC). The critical value for drought tolerance was SWC 6% while for the control plants the critical SWC was 8%; a moderate level of SWC is 13% and the highest SWC for plant endurance was 18%. The method proved to be reliable and sensitive in the evaluation of drought tolerance for forest trees.     

Drought stress tolerance analysis of Populus ussuriensis clones with different ploidies

The selection of drought-tolerant plants is an important aspect of plant breeding.We studied physiological and biochemical mechanisms of different ploidies of Populus ussuriensis Kom.that relate to drought stress tolerance.We used a 5%(v/v)polyethylene glycol(PEG-6000)solution to simulate drought stress.We recorded leaf phenotypes including color,dry area and curl degree.We evaluated sequential variations in some drought stress tolerance-related physiological and biochemical indices and compared these among diploid clones(CK),triploid clones(T12)and tetraploid clones(F20).T12 leaves exhibited slightly more drought stress damage than CK and F20 leaves.CK leaves suffered the most severe drought stress damage.The physiological and biochemical indices of the different ploidies differed significantly 12 days after drought stress treatment.The activities of superoxide dismutase,peroxidase,catalase and proline in the triploid(T12)leaves were the highest.The relative electric conductivity and malondialdehyde content of T12 leaves were the lowest.The index values of F20 were between those ofthe diploid and triploid.In consideration of these results,the drought resistance of the three different ploidies of P.ussuriensis can be ranked as T12>F20>CK.We speculate that the gene expression patterns of polyploid clones of poplar will change after genome doubling and that some of the drought stress tolerance-related physiological and biochemical indices will be improved,resulting in greater drought tolerance of polyploid clones.     

Identification of <Emphasis Type="Italic">CpTI</Emphasis> gene integration for 2-year-old transgenic poplars at DNA level

The putative transgenic hybrid triploid poplars [(P. tomentosa × P. bolleana) × P. tomentosa] with CpTI gene have been outplanted in test field for 2 years. Although the authors’ previous studies have proved that they are highly resistant to 3 species of poplar-threatening insect pests and contain high content of CpTI protein in foliage, incorporation status of foreign CpTI gene in poplar genome is uncertain. In this present study, the incorporation of foreign CpTI gene in genome of 5 transgenic poplars was confirmed by PCR and Southern blotting analysis. DNA amplification showed that there were clear DNA bands of about 450bp specific to CpTI gene in transgenic lanes, while no corresponding band in non-transgenic lane was observed. Correspondingly, clear DNA hybridization signals and no signal were exhibited on film for DNA Southern blotting analysis in transgenic lanes and non-transgenic lane, respectively, which further confirmed the stable integration of foreign CpTI gene in genome of 2-year-old transgenic poplar.     

杨树伐桩嫁接与植苗林的生长及碳储量的比较

The leaf nutrient, biomass and carbon reserve of 9-year-old Populus×euramericana cv. ‘74/76’ stump grafting stand with 26-year-old P.×xiaozhuanica W.Y.Hsu et Liang cv.‘Popularis’as rootstock, 26-year-old P.×xiaozhuanica W.Y.Hsu et Liang cv.‘Popularis’ and 9-year-old P.×euramericana cv. ‘74/76’ planting stands were compared. The result showed that the average increment and the annual average increment of stump grafting stand were greater than the two planting stands. The total nitrogen content in leaves of stump grafting stand was the highest, and it was 1.26 times and 1.05 times as much as that of the planting stands of Populus×xiaozhuanica W.Y.Hsu et Liang cv.‘Popularis’ and P.×euramericana cv. ‘74/76’ separately. The total phosphorus content of stump grafting stand was 1.3 times as much as that of planting stand of 26-year-old P.×xiaozhuanica W.Y.Hsu et Liang cv.‘Popularis’, but there was little difference between stump graftin stand and planting stand of 9-year-old P.×euramericana cv. ‘74/76’. The total potassium of stump grafting stand was 46.6% and 95.8% lesser than that of the planting stands of P.×xiaozhuanica W.Y.Hsu et Liang cv.‘Popularis’ and P.×euramericana cv. ‘74/76’. The biomass of aerial parts and the biomasses of trunk, branches and leaves of stump grafting stand were 1.84, 1.80, 1.76, and 3.00 times as much as that of planting stand of P.×euramericana cv. ‘74/76’ separately. The total root biomass of the stump grafting stand was 1.42 times and 7.68 times as much as that of planting stands of 26-year-old P.×xiaozhuanica W.Y.Hsu et Liang cv.‘Popularis’ and 9-year-old P.×euramericana cv. ‘74/76’. There was an obvious advantage in carbon reserve from all the parts of the stump grafting stand and the carbon sequestration capacity was the highest, which was 2.66 times and 1.62 times as much as that of planting stands of 26-year-old P.×xiaozhuanica W.Y.Hsu et Liang cv.‘Popularis’ and 9-year-old P.×euramericana cv. ‘74/76’. These showed that the stump grafting stand had more advantages than planting stands.     

Successful <Emphasis Type="Italic">Agrobacterium</Emphasis>-mediated transformation of <Emphasis Type="Italic">Populus tomentosa</Emphasis> with apple <Emphasis Type="Italic">SPDS</Emphasis> gene

The problem of salinized soils has become one of the most serious constraints to agricultural and forest productivity. With the purpose of enhancing salt stress tolerance of Populus tomentosa, we transformed this tree species with spermidine synthase （SPDS） genes derived from an apple by an Agrobacterium-mediated method. Four transgenic clones were confu＇med by PCR and Southern blot analysis. As well, the expression of introduced SPDS genes was analyzed by real-time quantitative PCR.     

High level expression of glucose-6-phosphate dehydrogenase gene <Emphasis Type="Italic">PsG6PDH</Emphasis> from <Emphasis Type="Italic">Populus suaveolens</Emphasis> in <Emphasis Type="Italic">E. coli</Emphasis>

In order to investigate the functions of the gene PsG6PDH and the mechanisms underlying freezing tolerance of Populus suaveolens, the recombinant expression vector pET-G (pET30a-G6PDH), which contained full encoding region of PsG6PDH gene, was established. The recombinant was identified by lawn-PCR and double enzyme digestion and then transformed into expression host XA90 and induced by isopropyl-â-D-thiogalactoside (IPTG) to express 100 kD polypeptide of G6PDH fusion protein. The results showed that the expressed amount of the fusion protein culminated after 1 mmol&#8226;L^–1 IPTG treatment for 4 h and that pET-G product was predominately soluble and not extra-cellular secreting.     

Verification of the resistance of a LEA gene from Tamarix expression in Saccharomyces cerevisiae to abiotic stresses

The role of late embryogenesis abundant （LEA） proteins in stress tolerance was examined by using a yeast expression system. LEA protein tolerance to the abotic stresses in plants involved in salt, drought and freezing stresses and additional tolerance to heat, NaHCO3 （salt-alkali） and ultraviolet radiation was also investigated. The transgenic yeast harboring the Tamarix LEA gene （DQ663481） was generated under the control of inducible GAL promoter （pYES2 vector）, yeast cells transformed with pYES2 empty vector were also generated as a control. Stress tolerance tests showed that LEA yeast transformants exhibited a higher survival rates than the control transformants under high temperature, NaHCO3, ultraviolet radiation, salt （NaCl）, drought and freezing, indicating that the LEA gene is tolerant to these abiotic stresses. These results suggest that the LEA gene is resistant to a wider repertoire of stresses and may play a common role in plant acclimation to the examined stress conditions.     

利用蒲螨和管氏肿腿蜂防治双条杉天牛幼虫

Pyemotes sp. and Scleroderma guani were used to control the larvae of Semanotus bifasciatus. The results as follows: (1) the suitable control time of Pyemotes sp.was from early April to early May. The reason was that in this time, the larvae of S. bifasciatus were young and not bored into xylem that was so easy to be parasitized by Pyemotes sp. With the parasitic rate 80% in average. Indoor experiment showed that the Pyemotes sp. had the same tropism selection to every instar for the host, but the parasitic rate was higher in low instar than other instars.(2)Using Scleroderma guani to control S. bjfasciatus, the suitable time was from mid May to mid June, the optimal ratio of host and parasite was 1∶ 1. Indoor experiment showed that Scleroderma guani had strong tropism toward middle-aged and elder larvae, the parasite linger around them and choose suitable position on hosts where they could lay eggs successfully. This behavior showed that Scleroderma guani had strict selectivity for their hosts. In experiment, the authors found a phenomenon that in one trap wood, both the Pyemotes sp. and Scleroderma guani could be found at the same time, they themselves did not affect each other and otherwise they could increase the parasitic rate for host. So, the authors suggested that in practice to control pests especially for boring pests, the Scleroderma guani and Pyemotes sp could be used at the same time.     

Over expression of TaFer gene from Tamarix androssowii improves iron and drought tolerance in transgenic Populus tomentosa

Ferritin, a universal intracellular protein, can store large amounts of iron and improve plant resistance to abiotic and biotic stress. In this study, a ferritin gene(TaFer) from Tamarix androssowii Litv. was transferred into Populus tomentosa Carr. cv 'BJR01' via Agrobacterium. Six independent transgenic lines were obtained with a tolerance to kanamycin and three were randomly selected for further analysis. The PCR and RT-PCR results indicate that the TaFer gene had been integrated into the poplar genome. The effect of the gene on abiotic stress tolerance was tested, and the results show that transgenic plants improve growth, had higher chlorophyll and lower MDA contents, and higher relative electrical conductivity,fewer changes of SOD and POD activities, higher iron content, higher root ferric reductase activity and lower levels of ROS accumulation and cell death in response to drought, Fe-insufficient or Fe-excess tolerance. These results indicate that the TaFer gene can improve abiotic stress tolerance in transgenic Populus tomentosa.     

Morphogenetic responses of Populus alba L. under salt stress

The morphogenetic responses to salt stress of Tunisian Populus alba clones were studied in order to promote their plantation in damaged saline areas.One year-old plants of three P.alba clones(MA-104,MA-195 and OG)were subjected to progressive salt stress by irrigation during two consecutive years.The plants were grown in a nursery,inside plastic receptacles containing sandy soil and were irrigated with tap water(control)or 3–6 g/l NaCl solution.During this study,leaf epinasty,elongation rate,vigor,internode length,plant architecture,and number of buds were evaluated.Test clone response was highly dependent on the applied treatment and degree of accommodation.The most pronounced alterations were induced under 6g/l of NaCl treatment including leaf epinasty,leaf elongation rate delay,vigor decrease,internode length shortening,and morphogenetic modifications.These responses were less noticeable in the MA-104 clone with respect to the two other clones.The salt effect induced a delay in the leaf elongation rate on the MA-195 and OG clones leading to an early leaf maturity.The vigour and internode length of the MA-104 clone was less affected than the other clones.The OG clone was the most salt-sensitive thus,it developed shorter branches and more buds number than MA-195 and MA-104.The effect of long-term salt stress was to induce early flowering of the P.alba clones which suggests that mechanism of salt accommodation could be developed.     

Application of Molecular Marking Technologies in Heredity Breeding of Eucalyptus

Two clonal trial stands of Chinese Fir （Cunninghamia lanceolata） were used in this study, one was 19-year-old stand which included 38 clones, and the other was 17-year-old stand including 102 clones.The statistical analyses showed that there were very significant genetic variations in height, DBH,volume and ratio of heartwood(R_hw),wood basic density（ρ_b ） of the clones in the two stands. The repeatability of clones was in median to high level,and the genetic CV was different over the all five traits.There were very significant phenotypic and genetic correlations among height,DBH and volume,and negative correlations among growth, R_hw andρ_b.The selection method experiment indicated that index selection could improve volume, R_hw andρ_b,showing synthetically superior selection effects compared to any individual trait selection methods.     

杂交常绿卫矛与其父母本的抗寒生理特性比较

Euonymus evergreen hybrid(1#, 2#, 3#) and their parents (Euonymus japonicus‘Zhuzi’, E. kiautschovicus) were studied to test whether their cold resistance were stronger than their parents in the winter. The result showed The semi-lethal temperatures (LT₅₀) were assessed separately by electrolyte leakage, the result showed that the LT₅₀ of 1#, 2#, 3#, E. japonicus‘Zhuzi’ and E. kiautschovicus were 36℃, 32.1℃, -28.5℃, -26.9℃, and -28℃ respectively. The activities of superoxide dismutase (SOD), peroxidase (POD) in 3# were higher than that of the others, but there was no significant difference among the other varieties. The contents of proline in leaves of E. japonicus‘Zhuzi’ were higher than that of Euonymus evergreen hybrids and E. kiautschovicus. The present study indicates that the hybrid evergreen euonymus and their parents had different mechanisms of cold-resistance.     

4种山茶花营养成分及有害元素含量分析

The nutritional components and poisonous elements in flowers of Camellia chekiangoleosa, C. polydonta, C. semiserrata and C. azalea were studied. The results indicated that the soluble sugar content of the four Camellia flowers had no difference. The maximum protein content was 82.9 g·kg^-1, detected in the flower of C. semiserrata. And the minimum protein content was 56.8 g·kg^-1, found in C. azalea. Vitamin C and β-carotene contents in C. azalea and C. semiserrata were higher, and in C. polydonta and C. chekiangoleosa were lower. The total amino acid contents of the four Camellia were 68.41、67.26、50.77、66.87 g·kg^-1 respectively, the essential amino acid contents were 33.86、32.70、27.21、33.79 g·kg^-1 respectively, and the proportion of essential amino acid in total amino acid of the four Camellia were 49.50%, 48.62%, 53.59% and 50.53% respectively. The contents of Ca, Mn and Cu in C. chekiangoleosa were the highest. Zn and Se contents were the maximum and Ca was the minimum in C. polydonta. The contents of Fe and Se were the lowest in C. semiserrata. Fe content were the maximum and Mn, Zn and Cu the minimum in C. azalea. The trace element contents in the flower of the four Camellia species followed the order of Mn, Fe, Zn, Cu, Se, but the content of Fe was higher than that of Mn in C. azalea. The contents of Pb, Cd, As and Hg of the four Camellia flower were all lower than the permissible amount.     

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&#8226;L^–1 6-BA benefitted callus proliferation and 0.25 mg&#8226;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&#8226;L^–1.     

Effects of soil moisture regimes on growth and photosynthesis of the riparian plant Bolboschoenus planiculmis

Plants distributed in riparian regions experience frequent episodes of flooding and drought between years, and hence, riparian plants need to be floodand drought-tolerant. Riparian plants possess various traits to survive flooding, while their sensitivity to drought has received less attention. To investigate the growth and photosynthetic responses of a riparian species (Bolboschoenus planiculmis) to flooding and drought, plants of this species were subjected to 60-d flooding or drought stress under greenhouse conditions. Growth and photosynthetic traits were measured at the end of the treatments. As well, we determined the efficiency of photosynthetic apparatus in mature leaves. Plants of B. planiculmis adequately adjusted their growth and photosynthetic traits under both flooding and drought conditions. Flooding did not affect the above-ground growth of B. planiculmis. Increased growth of roots and rhizomes and the generation of new tubers suggested a high ability of below-ground lateral growth by capturing resources under flooding conditions. Enhanced photosynthetic capacity, retained leaf pigment concentrations and chlorophyll a fluorescence capacity indicated photosynthetic adaptation to flooding. In contrast, drought significantly decreased the above-ground growth of B. planiculmis, especially the leaves, thereby minimizing water loss due to transpiration. Its increased root to shoot ratio and "phalanx" asexual propagation pattern might enhance soil water uptake ability. Although the functional leaves of B. planiculmis could retain their leaf pigment concentrations, as well as photosynthesis and chlorophyll a fluorescence, the total biomass of plants decreased, which may be a consequence of the reduced leaf area, suggesting adverse effects by drought. Therefore, both growth and photosynthetic responses of B. planiculmis are likely to contribute to the ability of this species to thrive in riparian regions, but remain susceptive to drought.     

不同品种油茶嫁接苗根系生长动态研究

The growth dynamics of root system and aboveground of the grafted seedlings of different Camellia oleifera cultivars are studied by skeleton method and root scanning. The results of 8-months’ observation on grafted seedlings of 5 C. oleifera cultivars showed that: the growth of aerial parts and root of the C. oleifera seedlings were not entirely in sync, during the time of low temperature from November to February, the root system still in sustained growth though the aerial parts of the 5 cultivars grew slowly. Remarkable differences were existed in root length, root surface area, root volume, root biomass and other indicators after 11-months’ growth. The mean value of total length of root-system was up to 243.3 cm and 143.4 cm for the minimum, the mean value of total root surface area was up to 50.2 cm² and 31.7 cm² for the minimum, the mean value of root volume of the C. oleifera seedlings was up to 0.843 cm³ and 0.578 cm³ for the minimum which lived 11 months after grafting. All the root length, root surface area, root volume of the 5 cultivars have remarkable differences.     

Transcriptome analysis reveals dynamic changes in the salt stress response in Salix

Soil salinization is a serious ecological problem worldwide and information regarding the salt tolerance mechanisms of Salix is scarce. To elucidate the dynamic changes in the molecular mechanisms of Salix under salt stress, we generated gene expression profiles and examined changes in the expression of those genes. RNA-Seq was used to produce six c DNA libraries constructed from the leaves of Salix 9 jiangsuensis CL ‘J2345' treated with Na Cl for 0, 2, 6, 12, 24 and 48 h. In total, 249 million clean reads were assembled into 12,739 unigenes, all of which were clustered into 10 profiles based on their temporal expression patterns. KEGG analysis revealed that as an early defense response, the biosynthesis pathways of cutin,suberin and wax, which are involved in cell wall structure,were activated beginning at 2 h. The expression of secondary metabolism genes, including those involved in the phenylpropanoid, flavonoid, stilbenoid, diarylheptanoidand gingerol pathways, peaked at 6 h and 24 h; the upregulated genes were mainly involved in plant hormone pathways and beta-alanine, galactose and betalain metabolism. We identified roles of key phytohormones and found ETH to be the major signaling molecule activating TFs at 12 h; ETH, ABA, IAA and SA were the key molecules at 24 h. Moreover, we found that the upregulated genes were associated with elevated levels of amino acids, sucrose, inositol, stress proteins and ROS-scavenging enzymes, contributing to the maintenance of water balance. This research constitutes the first detailed analysis of salt stress-related mechanisms in Salix and identifies potential targets for genetic manipulation to improve yields.