Determination of drought tolerance using root activities in Robinia pseudoacada ''''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.     

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.     

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.     

转基因多抗杨3号新品种选育

Populus euramericana‘MR3’is a transgenic clone transformed with five foreign genes (JERF36, SacB, vgb, and BtCry3A+ OC-1) via genetic engineering approach using Populus ×euramericana‘Guariento’as the receptor. This variety inherits the features of fast-growing and broad adaptation that the receptor possessed and further enhances the tolerance to various stresses including drought, salt, flooding, and insects feeding. The experiments in greenhouse for stress tolerance assays indicated that the expression of foreign genes led to improved tolerance to drought, salt, flooding, and insects (mainly Plagiodera versicolora). Field trial in an area with salinized soil in Shandong Province showed a 7.77% increase in stem volume than that of the control clones of 5-year-old saplings. Another field trial at Linghai in Liaoning Province indicated a 15.41% increase in stem volume than that of the control clones of 2-year-old. In the experiments in greenhouse, laboratory, and field, the P. euramericana‘MR3’showed enhanced characteristics such as fast-growing, higher tolerance to drought, salt, insects, and flooding stress, and is expected to be one of the ideal new poplar varieties for wood industry and ecosystem conservation in North China.     

Regeneration and gene transformation systems of Robima 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 concentration of it was 8 mg·L-1.     

Osmoregulation mechanism of drought stress and genetic engineering strategies for improving drought resistance in plants

Drought, one of the main adverse environmental factors, obviously affected plant growth and development. Many adaptive strategies have been developed in plants for coping with drought or water stress, among which osmoregulation is one of the important factors of plant drought tolerance. Many substances play important roles in plant osmoregulation for drought resistance, including proline, glycine betaine, Lea proteins and soluble sugars such as levan, trehalose, sucrose, etc. The osmoregulation mechanism and the genetic engineering of plant drought-tolerance are reviewed in this paper.     

Drought tolerance of Acacia karroo, a native tree species to South Africa, in Beijing

Acacia karroo is an indigenous, strong drought-tolerant and widely planted tree in South Africa. In order to match the plant with its adaptation to forest sites in China, it is important to know the drought-tolerance of this species under Chinese forest site conditions. Our study was carried out in cinnamon soils in Beijing and with Platycladus orientalis, the most drought-tolerant tree in Beijing, as control. Physiological parameters of young plants of the trees were measured and divided into three groups in response to soil drying. The first group included root activity, ABA content in roots and peroxidase (POD) activity in leaves. Their values changed similarly to those of P. orientalis and dramatically increased to maximum values when the soil water content (SWC) decreased from its normal level of 20% to 7.6% where the three indices decreased dramatically when SWC was reduced further from 7.6%. The second group of parameters included the contents of proline, soluble protein and malondialdehyde (MDA). Their values showed greater variation than those of P. orientalis and significantly increased when SWC decreased from 20% in response to soil drying. However, the proline and soluble protein contents fluctuated in A. karroo at a lower level than those in P. orientalis. In contrast, the content of MDA in A. karroo was much higher than that in P. orientalis which almost did not increase in the drying soil. The third group of parameters consisted of soluble sugar, superoxide dismutase (SOD) and catalase (CAT). These parameters showed greater diversity and fluctuations among the three varieties of A. karroo compared with the control in response to soil drying. These results suggest that A. karroo is characterized as a strong, drought tolerant species in most of physiological parameters but weaker than P. orientalis in membrane lipid antioxidation.     

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.     

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.     

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.     

裳凤蝶污斑亚种的生物学与规模化饲养的初步研究

The largest and most precious Chinese butterfly Troides helena spilotia Rothschild is occurring all year round with overlapping 6 generations in Hainan Jianfengling forest each year. The appropriate growth temperature for it is 25－30 ℃, and the humidity is 70%－80%. Large-scale artificial breeding of Troides helena spilotia Rothschild need to comply with the requirements of their breeding rooms by planting host plants for their eggs. Dilution of the concentration of honey and water is 10%－15% in summer and 5% in winter; the eggs should be moisturizing when collecting indoor and outdoor. The larvae should be transferred to the host plants planted in the field with net bags; and transfer them to different host plants timely according to their instars and food intake; the indoor population density should be controlled. Due to the 4－5 instars larvae will ingest a great deal of food, they need to be fed 3 times a day in the morning, afternoon and the evening. The ways to suspend the pupa when feeding indoor and outdoor vary according to the breeding facilities. The pupa should be collected and put in the emergence room and hung them up according to their natural shape. The suitable temperature for it is 25－28 ℃, and humidity is 60%－80%.     

Expression of a carrot 36 kD antifreeze protein gene improves cold stress tolerance in transgenic tobacco

Antifreeze proteins （AFPs） enable organisms to survive under cold conditions, and have great potential in improving cold tolerance of cold-sensitive plants, In order to determine whether expression of the carrot 36 kD antifreeze protein gene confers improved cold-resistant properties to plant tissues, we tried to obtain transgenic tobacco plants which expressed the antifreeze protein. Cold, salt, and drought induced promoter Prd29A was cloned using PCR from Arabidopsis. Two plant expression vectors based on pBI121 were constructed with CaMV35S：AFP and Prd29A：AFP. Tobacco plantlets were transformed by Agrobacterium-medicated transformation. PCR and Southern blotting demonstrated that the carrot 36 kD afp gene was successfully integrated into the genomes of transformed plantlets. The expression of the afp gene in transgenic plants led to improved tolerance to cold stress. However, the use of the strong constitutive 35S cauliflower mosaic virus （CaMV） promoter to drive expression of afp also resulted in growth retardation under normal growing conditions. In contrast, the expression of afp driven by the stress-inducible Prd29A promoter from Arabidopsis gave rise to minimal effects on plant growth while providing an increased tolerance to cold stress condition （2℃）. The results demonstrated the prospect of using Prd29A-AFP transgenic plants in cold-stressed conditions that will in turn benefit agriculture.     

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.     

Capacity of Stem Water Conductivity for Two Eucalyptus (Eucalyptus urophylla) Plantations in South China

The sap flux density (SFD) was used as a measure of water capacity through stems of eucalyptus in this paper. It was found that daily SFD increased with daily vapor pressure deficit (VPD) in nonlinear regression Y = AX³ + BX² + CX + D (R² = 0.702 1, n = 135, p = 0.01) at both sites (Hetou and Jijia) in the Leizhou Peninsula, Guangdong Province, China, where Y was daily SFD, X was mean daily VPD, A, B, C, D were constants. But extremely high VPD might limit stem water conductivity. The environmental factors, such as air vapor pressure deficit (VPD), solar radiation (RAD), etc., were the main determinants of SFD for E. urophylla plantations. The upper threshold of diurnal SFD was 51.55–55.65 mL&#8226;cm^–2&#8226;h^–1 under the selected extremely high environmental conditions.     

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 ofβ-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 blotting. 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.     

Photosynthetic stimulation of saplings by the interaction of CO_2 and water stress

Climate change necessitates research into interactions between elevated carbon dioxide(CO2)concentrations and drought on plant photosynthetic physiology.This study describes the physiological properties of Platycladus orientalis(Chinese thuja)and Quercus variabilis(Chinese cork oak)saplings cultivated through orthogonal treatments of four CO2 concentrations combined with five soil volumetric water contents(SWC).It highlights the differences between the interactive effects from the treatments.Water stress had little effect on photosynthetic traits until the soil volumetric water contents exceeded 70–80 or 100%.Similar variations in carbon-13 isotope abundance(δ^13C)of water soluble compounds(δ^13CWSC)extracted from leaves of two species have been observed.Whether soil volumetric water contents exceeded or fell below the water threshold values(70–80%of field capacity for P.orientalis and 100%of field capacity for Q.variabilis),instantaneous water use efficiency decreased.Elevated carbon dioxide could increase iWUE and enhance drought tolerance,depending on stimulating net photosynthetic rates and declining stomatal conductance and transpiration rates.Augmenting either drought,excess water,or ambient carbon dioxide could alleviate the physiological inhibition caused by the stresses described above.     

Response of soil respiration to a severe drought in Chinese Eucalyptus plantations

Extreme droughts can adversely affect the dynamics of soil respiration in tree plantations. We used a severe drought in southwestern China as a case study to estimate the effects of drought on temporal variations in soil respiration in a plantation of Eucalyptus globulus. We documented a clear seasonal pattern in soil respiration with the highest values(100.9 mg C–CO_2 m~(-2)h~(-1)) recorded in June and the lowest values(28.7 mg C–CO_2 m~(-2)h~(-1)) in January. The variation in soil respiration was closely associated with the dynamics of soil water driven by the drought. Soil respiration was nearly twice as great in the wet seasons as in the dry seasons. Soil water content accounted for 83–91% of variation in soil respiration, while a combined soil water and soil temperature model explained 90–99% of the variation in soil respiration. Soil water had pronounced effects on soil respiration at the moisture threshold of 6–10%. Soil water was strongly related to changes in soil parameters(i.e., bulk density, p H,soil organic carbon, and available nitrogen). These strongly influenced seasonal variation in soil respiration. We found that soil respiration was strongly suppressed by severe drought. Drought resulted in a shortage of soil water which reduced formation of soil organic carbon, impacted soil acid–base properties and soil texture, and affected soil nutrient availability.     

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.     

Review on Factors Affecting Plant Competition

The competition refers to the interactions of two or more individuals which compete for resources,and is a key process of plant populations and communities.A comprehensive understanding of plant competition is necessary to predict the responses of ecological systems to environmental changes.The inside factors affecting competition include plant density,biological factors and the biomass.The environmental factors related to competition include light,soil moisture,soil nutrient and altitude.High density affects plant competition by controlling plant growth.Allelochemicals produced by plants directly affect their neighbors. Trees and grasses may compete for resources where their root systems overlap.Herbivorous animals chose to eat some plants,resulting in reduction of competition ability of the plants.Plants make morphological shifts and alter the competitive ability between above-ground and below-ground parts in response to the environmental changes.When shade- tolerant species are absent in mixed forest,less shade tolerant species could maintain their dominance for a long period.In contrast,mixed forest with tolerant species would reduce the dominance of less-tolerant species,maintain and probably increase the dominance through the decline of less-tolerant species.Smaller plants should have longer survival time due to their smaller total water requirements when water is scarce,and the longer survival of plants with higher root allocation could be due to a relative reduction in transpiring surface per unit root length.In nutrient-deficient soils,plants are stressed directly by the lack of adequate nutrients and competitive interactions may be controlled by a plant’s ability to efficiently take up available nutrients.In nutrient-sufficient soils,plants with the highest maximum growth rates may well be the superior competitors.Intensity of light competition declines with altitude.The future research about competition will focus on large scale,long term and multi-population research,the multi-disciplinary cooperat