Evaluation of cold resistance of ornamental species for planting as urban rooftop greening

Rooftop gardening or green roof establishment, one of the methods of afforestation for urban greening, has many benefits for the environment, the economy and urban landscapes. Various environmental stresses including heat, strong wind, sunshine and cold prevent most plants from growing well in extensive green roof systems. For the establishment of urban rooftop gardening in Beijing, we evaluated 13 ornamental plant species [Berberis thunbergii ‘Atropurea Nana’, Euonymus fortune （Turcz.） Hand., Viburnum dilatatum Thunb., Cotinus coggygria Scop., Swida alba Opiz., Weigela florida （Bunge.） A. DC., Jasminum nudiflorum Lindl., Euonymus japonicus Thunb., Euonymus japonicus ‘Cuzhi’, Buddleja davidii Franch., Pyracantha fortuneana （Maxim.） Li., Ligustrum lucidum Ait. and Spiraea salicifolia L.] for greening of flat rooftops. We obtained three indices concerning cold resistance, i.e., electrical conductivity of leaves, amounts of soluble sugars and amounts of free proline, providing a theoretical basis for selection of plants fit for roof and balcony greening. The results show that the six species P. fortuneana （Maxim.） Li., B. thunbergii ‘Atropurea Nana’, J. nudiflorum Lindl., E. fortune （Turcz.） Hand., E. japonicus Thunb. and E. japonicus ‘Cuzhi’ are excellent plants for rooftop gardening, judging from their strong ability in cold resistance. Plants such as L. lucidum Ait., C. coggygria Scop. and W. florida （Bunge） A. DC. showed a medium ability in cold resistance. However, these species can be used in roof and balcony greening after intensive management or acclimatization of variety. The ability to resist cold of plants such as W. florida （Bunge） A. DC., S. salicifolia L., V. rhytidophyllum and S. alba Opiz. （Cornus alba L.） was found to be rather weak. Therefore, cold-proof measures are recommended when using these species.

Cross Breeding of Populus and Its Hybrids for Cold Resistance

Populus tomentosa was crossed with P.tremuloidis, P.grandidentata, P.alba×P.grandidentata and P.alba×Ulmuspumila 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 2744 hybrid seedlings were obtained. MX4×P.grandidentata （G-1-58）, MX3×P.tremuloidis （T-44-60）, MX2×P.tremuloidis （1-13-87-37） 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.

Molecular Cloning of a Cytosolic G6PDH Gene from Populus Suaveolens and Its Expression to Improve the Cold Resistance of Tobacco Plants

Glucose-6-phosphate dehydrogenase (G6PDH,EC 1.1.1.49) is the first and main regulated enzyme of oxidative pentose phosphate pathway (OPPP),catalyzing the conversion of glucose-6-phosphate to 6-phospho-gluconolactone and playing important roles in the growth and development of plants. It is preciously reported that the enhancement of freezing resistance of Populus suaveolenscuttings is clear related to the distinct increase in cytosolic G6PDH activity. Here,a 1697 bp cDNA fragment (PsG6PDH) is amplified by RT-PCR from cold-induced total RNA of the freezing-tolerant P. suaveolens. A sequence analysis showed that PsG6PDH coding region had 1 530 bp and encoded 510 predicted amino acid residues. Genomic Southern analysis revealed that the isoform is encoded by a few copies of the gene in the poplar genome. The cloned gene PsG6PDHis cloned into binary vector pBI121 and used to transform tobacco. PCR and Southern blotting results verified integration of this gene into the genome of tobacco. Moreover,cold treatment experiments and membrane defense enzymeactivity analysis confirmed that overexpression of the PsG6PDHgene could enhance the tolerance to cold or frigid stresses in transgenic plants.

A Review of the Methods of Identifying and Testing Plant Cold Resistance

Low-tempemture stress is a major abiotic stress influencing plant growth, economic yield and quality worldwide. Precise and reliable identification and test of plant cold resistance is a foundation for exploring low-temperature damages, cold resistance mechanism of plants, cultivating and innovating quality cold-re- sistant germplasm resources, which is rather important for woody plants because their perennial and large-sized characteristics. Generally, most researches on plant cold-resistant physiology focused on identification and utilization of cold resistance worldwide, and a few on test methods of cold resistance, but cold resistance mechanism has been less reported. Therefore, researches on methods of identifying and testing plant resistance have achieved considerable progress, accumulated much experience and formed characteristics gradually in terms of methodology. Current methods of identifying plant cold resistance include open field/outdoor culti- vation evaluation, cold and frost damage investigation, simulated cold weather, and mathematical model forecast. And the methods of testing plant cold resistance include growth condition test, frozen index investigation, electrolyte leakage, bioelectrical impedance mapping and so on. Synthesizing the methods of identifying and testing plant cold resistance should be a key matter in the present and future researches and application of plant cold resistance physiology, improvement and in- novation of the identification and test will be an important direction of the future researches. Identification and test of plant cold resistance based on genomies, meta- bonomics and proteomics should be the new trend of the researches on physiology and ecology of plant cold resistance.

Physiological mechanisms of resistance to cold stress associated with 10 elite apple rootstocks

A study was conducted in attempting to identify the cold-resistant apple rootstocks and to establish a comprehensive evaluation system. In this study, 10 elite apple dwarfing rootstocks(GM256, JM7, M26, M7, SC1, SH1, SH38, SH6, M9, and T337) were employed for the experiment and the following parameters were investigated under different low temperature stress conditions(0, –15, –20, –25, –30, and –35°C): the changes of the relative electrical conductivity(REC), anthocyanin content, protein content, soluble sugar content, soluble starch content, proline content, malondialdehyde(MDA) content, superoxide dismutase(SOD) activity, and peroxidase(POD) activity of the dormant branches. The inflection temperature that could represent the plant tissue semi-lethal temperature(LT) was obtained by the measurements of REC. The LTwas used to evaluate eight other indices. The results showed that there was no significant correlation between LTand POD activity as well as between the soluble sugar, protein and proline contents at 0 and –15°C. Soluble starch content at 0 and –15°C and anthocyanin content at –15–(–30)°C were significantly but negatively correlated to the LT50 and the MDA content at 0–(–20)°C was significantly positively correlated to the LT. Statistical analysis based on principal component analysis and LT50 showed that cold resistant apple rootstocks in the decreasing order from high to low as GM256, SH6, SH38, SH1, SC1, M26, M7, JM7, T337, and M9.

Comparison of transcriptome and metabolome analysis revealed differences in cold resistant metabolic pathways in different apple cultivars under low temperature stress

Freezing injury in winter is an important abiotic stress that seriously affects plant growth and development.Deciduous fruit trees resist freezing injury by inducing dormancy.However,different cultivars of the same species have different cold resistance strategies.Little is known about the molecular mechanism of apple trees in response to freezing injury during winter dormancy.Therefore,in this study,1-year-old branches of the cold-resistant cultivar‘Hanfu’(HF)and the cold-sensitive cultivar‘Changfuji No.2’(CF)were used to explore their cold resistance through physiological,biochemical,transcriptomics,and metabolomics analyses.Combining physiological and biochemical data,we found that HF had a stronger osmotic regulation ability and antioxidant enzyme activity than CF,as well as stronger cold resistance.The functional enrichment analysis showed that both cultivars were significantly enriched in pathways related to signal transduction,hormone regulation,and sugar metabolism under freezing stress.In addition,the differentially expressed genes(DEGs)encoding galactinol synthase,raffinose synthase,and stachyose synthetase in raffinose family oligosaccharides(RFOs)metabolic pathways were upregulated in HF,and raffinose and stachyose were accumulated,while their contents in CF were lower.HF accumulated 4-aminobutyric acid,spermidine,and ascorbic acid to scavenge reactive oxygen species(ROS).While the contents of oxidized glutathione,vitamin C,glutathione,and spermidine in CF decreased under freezing stress,consequently,the ability to scavenge ROS was low.Furthermore,the transcription factors apetala 2/ethylene responsive factor(AP2/ERF)and WRKY were strongly induced under freezing stress.In summary,the difference in key metabolic components of HF and CF under freezing stress is the major factor affecting their difference in cold resistance.The obtained results deepen our understanding of the cold resistance mechanism in apple trees in response to freezing injury during dormancy.

Integrative transcriptomic and metabolomic analyses reveal the flavonoid biosynthesis of Pyrus hopeiensis flowers under cold stress

Low temperature is among the most restrictive factors to limit the yield and distribution of pear. Pyrus hopeiensis is a valuable wild resource.PCA showed that P. hopeiensis had strong cold resistance. In this study, the mRNA and metabolome sequencing of P. hopeiensis flower organs exposed to different low temperatures were performed to identify changes of genes and metabolites in response to low-temperature stress. A total of 4 851 differentially expressed genes(DEGs) were identified. Trend analysis showed that these DEGs were significantly enriched in profiles 19, 18, 7, 14, 1, 4 and 11. And the KEGG enrichment analysis showed that the DEGs in profile 18 were significantly enriched in flavone and flavonol biosynthesis. Besides, the expressed trends as well as GO and KEGG functional enrichment analyses of DEGs under cold and freezing stress showed significantly difference. Analyses of flavonoid-related pathways indicated that flavonoid structural genes had undergone significant changes. Correlation analysis showed that b HLH and MYB TFs may affect flavonoid biosynthesis by regulating structural gene expression. And PhMYB308 and PhMYB330 were likely candidate repressors of flavonoid biosynthesis by binding to a specific site in bHLH proteins. In total, 92 differentially accumulated metabolites(DAMs) were identified in P. hopeiensis flowers including 12 flavonoids. WGCNA results showed that coral 1, pink and brown 4 modules were closely associated with flavonoids and 11 MYBs and 15 bHLHs among the three modules may activate or inhibit the expression of 23 structural genes of flavonoid biosynthesis. Taken together, the results of this study provided a theoretical basis for further exploration of the molecular mechanisms of flavonoid biosynthesis and cold resistance of P. hopeiensis flower organs and our findings laid a foundation for further molecular breeding in cold-resistant pear varieties.

Development of Hybrid Rice Variety FY7206 with Blast Resistance Gene Pid3 and Cold Tolerance Gene Ctb1

Hybrid rice Fanyou 7206（FY7206）, derived from the cross between a sterile line Fanyuan A and a restorer line Fuhui 7206, was bred by the Rice Research Institute, Fujian Academy of Agricultural Sciences, China. FY7206 was characterized by moderate blast resistance, cold tolerance, as well as wide adaptability, and high yields. The blast resistance results indicated that the frequencies of blast races in race B, race C and the total resistance frequency for FY7206 were 95.5%, 100.0% and 97.2%, respectively. The disease resistance results showed that the leaf blast grade for FY7206 was level 1 and panicle blast was level 5. The indoor spray results indicated that FY7206 was resistant to 11 isolates of Magnorpathe oryzae. The blast resistance of FY7206 might be derived from the high expression of blast resistance gene Pid3. The results for simulated cold resistance in an artificial climate chamber indicated that the cold tolerance for FY7206 was moderate at the booting and flowering stages. The cold tolerance results also indicated that FY7206 could be tolerant to temperatures as low as 10 °C at the seedling stage. The q RT-PCR results showed that the expression of cold tolerance gene Ctb1 in FY7206 was relatively high. These results suggested that FY7206 is a hybrid indica rice variety with good comprehensive characteristics, including blast resistance and cold tolerance.

Physiological Changes and Cold Tolerance of Three Camphor Species During Natural Winter Temperature Fluctuations

The dynamic changes in the malondialdehyde（MDA）, superoxide dismutase（SOD）, soluble sugar, proline, and soluble protein contents, as well as the relative electrolyte conductivity and the corresponding cold resistance, of Cinnamomum bodinieri Level., C. camphora L., and C. caudiferum Kisterm were investigated during the winter months of October 2009 to April 2010. During the short period of temperature decline that lasted until mid-December, the changes in the relative electrolyte conductivity and MDA content with temperature were insignificant. In January, SOD activity continued to increase and then peaked as a result of rapid increases in soluble sugar, proline, soluble protein, as well as the inhibition of the relative electrolyte conductivity and decrease in MDA content. These physiological changes protected the camphor trees from cold damage during winter. From February to March, SOD activity and the soluble protein and proline contents increased with the increase in temperature. However, the relative electrolyte conductivity and MDA content decreased, indicating that the cell membrane damaged by low temperature was gradually being repaired. The cold dip in April led to slight increases in the relative electrolyte conductivity and MDA content. Using a fuzzy mathematics method, the cold resistance adaptability of the camphor trees was divided into three periods namely, the enhancement setting stage, the vigorous stage, and the reducing stage. The cold tolerance abilities were ranked as the following order： C. bodinieri Level〉C. camphora L.〉C. caudiferum Kisterm.

High-precision Thickness Setting Models for Titanium Alloy Plate Cold Rolling without Tension

Due to its highly favorable physical and chemical properties,titanium and titanium alloy are widely used in a variety of industries.Because of the low output of a single batch,plate cold rolling without tension is the most common rolling production method for titanium alloy.This method is lack of on-line thickness closed-loop control,with carefully thickness setting models for precision.A set of high-precision thickness setting models are proposed to suit the production method.Because of frequent variations in rolling specification,a model structural for the combination of analytical models and statistical models is adopted to replace the traditional self-learning method.The deformation resistance and friction factor,the primary factors which affect model precision,are considered as the objectives of statistical modeling.Firstly,the coefficient fitting of deformation resistance analytical model based on over-determined equations set is adopted.Additionally,a support vector machine（SVM）is applied to the modeling of the deformation resistance and friction factor.The setting models are applied to a 1450 plate-coiling mill for titanium alloy plate rolling,and then thickness precision is found consistently to be within 3%,exceeding the precision of traditional setting models with a self-learning method based on a large number of stable rolling data.Excellent application performance is obtained.The proposed research provides a set of high-precision thickness setting models which are well adapted to the characteristics of titanium alloy plate cold rolling without tension.

Low Cycle Fatigue and Strengthening Mechanism of Cold Extruded Large Diameter Internal Thread of Q460 Steel

large diameter internal thread of high-strength steel（LDITHSS） manufactured by traditional methods always has the problems of low accuracy and short life. Compared with traditional methods, the cold extrusion process is an effective means to realize higher accuracy and longer life. The low-cycle fatigue properties of LDITHSS are obtained by experiments, and the initiation and propagation of fatigue cracks are observed by scanning electron microscope（SEM）. Based on the mechanical properties, surface microstructure and residual stress, the strengthening mechanism of cold extruded large diameter internal thread（LDIT） is discussed. The results show that new grains or sub-grains can be formed on the surface of LDIT due to grain segmentation and grain refinement during cold extrusion. The fibrous structures appear as elongated and streamlined along the normal direction of the tooth surface which leads to residual compressive stress on the extruded surface. The maximum tension stress of LDIT after cold extrusion is found to be 192.55 k N. Under low stress cycling, the yield stress on thread increases, the propagation rate of crack reduces, the fatigue life is thus improved significantly with decreasing surface grain diameter and the average fatigue life increases to 45.539×10~3 cycle when the maximum applied load decreases to 120 k N. The low cycle fatigue and strengthening mechanism of cold extruded LDIT revealed by this research has significant importance to promote application of internal thread by cold extrusion processing.

山东茶树叶片解剖结构分析

本文描述了茶树叶片的形态及解剖结构研究意义和现状,比较了 51 个茶树品种、育种材料的解剖结构,分析比较了不同品种、育种材料以及不同地区间的抗寒性,为茶树的北方地区茶树引种栽培、新品种和抗寒类型的筛选提供了理论依据。

Effects of Low Temperature Stress and INA Bacteriaon Chlorophyll a Fluorescence Induction Kineticsin Young Fruit of Two Apricot Cultivars

Effects of low temperature and INA bacteria on the change of chlorophyll a fluorescence in young fruit from two apricot cultivars were investigated. Low temperature decreased the potential activity (Fv/ Fo), conversion efficiency of primary light energy (Fv/Fm)of PS II and photochemical quenching (qP) in young fruit of two apricot cultivars. Low temperature enhanced non-photochemical quenching qN, decreasing the quantum yield of photosynthetic electron transfer. The presence of ice nucleating active (INA) bacteria intensified the effects of low temperature, raised the injury temperature threshold from - 4℃ to - 2 - - 3℃. INA bacteria can be a factor to induce frost susceptibility of apricot fruit. The amount of damaged PS I activity center was related to apricot fruit size and cultivar.

Studies on Cloning and Transformation of CBF1 Gene of Maize Grass

[Objective]It is revealed whether the similar maize transcriptional activator in CBF1 gene is regulatory cold resistance gene to lay the foundation for breeding new transgenic Forage Maize Varieties with high cold resistance ability.[Methods]In the present paper,the transcriptional factor gene CBF1 was Successfully cloned by PCR from the leaves of Arabidopsis.The sequence was preliminarily analyzed and plant expression vector was constructed.Then with agrobacterium-mediated transgene technique,CBF1 gene was introduced into maize SAUMZ1.[Results]PCR assay revealed that the CBF1 gene was integrated in the maize grass SAUMZ1 genome.Under different low temperature treatment,the relative electrolyte leakage percentage of transgenic plant was lower than Control.[Conclusion] The results showed that the cold-resistance of maize grass SAUMZ1 enhanced after transforming CBF1 gene.

Analysis on Membrane Stability of Crown in Winter Wheat in Frigid Region in China

Four varieties of winter wheat with different return green rates were used to analyze the plasma membrane stability and the factors that affect winter wheat in the frigid region during winter. The removal of reactive oxygen species, the degree of plasma membrane impairment, water composition and content, and the changes in cell viability in the crowns, which contain the growing point, were studied during the period from cold acclimation to the deep freezing. The results showed that electrical conductivity which reflects the degree of plasma membrane damaged under low temperature was significantly correlated with the free water and the total water content. The malondialdehyde （MDA） content, which reflects the degree of membrane peroxidation, was very significantly correlated with superoxide dismutase （SOD）, peroxidase, and ascorbic acid. During the deep freezing period, the SOD activity and glutathione （GSH） content of the winter wheat varieties were relative to their cold resistance. During this period, the MDA stability, SOD, GSH, and the total water and the free water content might be used to identify the cold resistance of winter wheat varieties.

The Changes in Anti-oxidant Activity of Roots in Wintering Period under Single Sowing of Alfalfa and Mixed Sowing of Alfalfa and Bromus inermis Leyss

This paper aimed to study the change in MDA content and antioxidant enzyme activity of roots in wintering period under single sowing of alfalfa and mixed sowing of alfalfa and Bromus inermis Leyss.By the single sowing of 4 alfalfa varieties with different cold resistance and mixed sowing of alfalfa varieties and Bromus inermis Leyss,the changes in MDA content,SOD,POD and CAT activity of alfalfa roots throughout the wintering period were measured.Results indicated that after single sowing of alfalfa and mixed sowing of alfalfa and Bromus inermis Leyss,the MDA content of roots showed an up-down-up trend with temperature; the CAT activity increased with the decreasing temperature but decreased when the temperature rose in the spring of the following year; the SOD and POD activity showed an increasing trend with sharp decline in temperature,and decreased when the temperature continued to decline,but increased with alfalfa reviving in the following year.The enzymic activity of the same variety under mixed sowing was higher than under single sowing.The membership function was used for comprehensive evaluation of cold resistance,and the cold resistance under different treatments was in the order of Wega7F + Bromus inermis Leyss >Wega7F > Xunlu + Bromus inermis Leyss > Xunlu > Aohan + Bromus inermis Leyss > Aohan > WL319HQ + Bromus inermis Leyss > WL319HQ.This was of great significance to the study on cold resistant alfalfa breeding and cultivation in cold areas of northern China.

Cold acclimation increases Asian citrus psyllid Diaphorina citri (Hemiptera: Liviidae) survival during exposure to freezing temperatures

The Asian citrus psyllid,Diaphorina citri,is the vector of the pathogenic bacteria that causes Huanglongbing,the most devasting disease affecting citrus worldwide.As cultivation of citrus tends to expand northward,the tolerance of psyllids to freezing temperatures needs to be investigated.While mortality of D.citri to cold temperature has been previously studied,much less is known regarding the acclimation potential of psyllids to cold temperatures.We first evaluated cold resistance of D.citri depending on color morph and sex.Subsequently,we compared mortality of D.citri at−4°C for 10 hours between unacclimated psyllids,psyllids that were gradually acclimated to cold over 1 or 2 weeks,and psyllids intermittently acclimated also for 1 or 2 weeks.We did not find major differences in cold resistance between color morphs and sex.We found that after 1 week of gradual acclimation,the survival of psyllids at−4°C increased to 86%,compared to 20%in the control group.Survival did not increase after the second week of gradual acclimation.Although intermittent acclimation improved survival compared to the control group,it was less effective than gradual acclimation with a survival at 30%and 70%after 1 and 2 weeks of acclimation,respectively,although this difference with gradual acclimation was not significant at week 2.These data show that gradual cold acclimation allows D.citri to survive brief periods below freezing.It may serve as a mechanism responsible for increasing northern establishment of this pest.

Variation in adult stress resistance does not explain vulnerability to climate change in copper butterflies

Ongoing climate change is a major threat to biodiversity. However, although many species clearly suffer from ongoing climate change, others benefit from it, for exam- ple, by showing range expansions. However, which specific features determine a species＇ vulnerability to climate change？ Phenotypic plasticity, which has been described as the first line of defence against environmental change, may be of utmost importance here. Against this background, we here compare plasticity in stress tolerance in 3 copper butter- fly species, which differ arguably in their vulnerability to climate change. Specifically, we investigated heat, cold and desiccation resistance after acclimatization to different temper- atures in the adult stage. We demonstrate that acclimation at a higher temperature increased heat but decreased cold tolerance and desiccation resistance. Contrary to our predictions, species did not show pronounced variation in stress resistance, though plastic capacities in temperature stress resistance did vary across species. Overall, our results seemed to reflect population--rather than species-specific patterns. We conclude that the geographical ori- gin of the populations used should be considered even in comparative studies. However, our results suggest that, in the 3 species studied here, vulnerability to climate change is not in the first place determined by stress resistance in the adult stage. As entomological studies focus all too often on adults only, we argue that more research effort should be dedicated to other developmental stages when trying to understand insect responses to environmental change.