Genome size microscale divergence of <Emphasis Type="Italic">Cyclamen persicum</Emphasis> in Evolution Canyon,Israel

Using DAPI flow cytometry, we examined genome size divergence of the Persian violet, Cyclamen persicum (Primulaceae) (2n=48) on close opposite slopes of Evolution Canyon (EC), Mt. Carmel, Israel. The range of genome size variation detected among measured cyclamens was 6.41% in relation to the smallest measured DNA content. Our data on C. persicum at EC showed that local variability in the 2C-value exists. Significantly less DNA was recorded in plants growing in one station of the African savannah-like south-facing slope (AS) but not in the remaining two stations of the same slope. We were not able to reject the null hypothesis that there are no significant interslope differences in the genome size between the temperate European garrigue-like north-facing slope (ES) and the drier AS. In spite of the nonsignificant interslope trend for the higher genome size in C. persicum, the data-fusion (meta-analysis) test using correlations between C-values in C. persicum, and earlier studied carob tree (Ceratonia siliqua), trifoil (Lotus peregrinus) and a beetle (Oryzaephilus surinamensis) and their distribution along the aridity gradient indicates a positive relationship between drought and genome size at the microsite.     

基于流式细胞术的海菜花属植物基因组大小测定

利用改良的裂解液P1,以中国古代莲(Nelumbo nucifera Gaertn.Fruct.et Semin)为外标,采用流式细胞术(FCM)对海菜花属(Ottelia Pers.)6个代表性物种及3个存疑类群的基因组大小(C值)进行测定,并对海菜花属系统发育关系进行评估。结果显示:所测定的材料中,水菜花(Ottelia cordata(Wall.)Dandy)C值最小(6.759 pg),灌阳水车前(O.guanyangensis Z.Z.Li,S.Wu&Q.F.Wang)C值最大(12.929 pg);对基因组大小与该属系统发育树进行比较分析,结果发现该属植物基因组大小与系统发育关系具有一致性;对海菜花属3个存疑类群进行分子系统学研究,结果发现存疑类群与嵩明海菜花(Ottelia acuminata var.songmingensis Z.T.Jiang,H.Li&Z.L.Dao)及灌阳水车前的关系最近,而与水菜花的关系较远,这与基因组大小变异相一致。根据基因组大小进一步推测3个存疑类群很可能为二倍体。本研究结果可为海菜花属植物的系统学研究提供新资料,同时为该属植物基因组学研究提供基础数据。     

Quantification of nuclear DNA and G-C content in marine macroalgae by flow cytometry of isolated nuclei

Summary The amounts of nuclear DNA in ten species of seaweeds belonging to the Rhodophyceae, Phaeophyceae, and Chlorophyceae were determined by flow cytometric analysis of nuclei isolated from protoplasts. Genome size was determined from the fluorescence of the nuclei stained with ethidium bromide. The size of the nuclear genome ranged from 0.13 pg per cell in the 1 C population ofUlva rigida to 3.40 pg per cell in the 2 C population ofSphacelaria sp. GC% analysis was based on staining with either Hoechst 33342 or mithramycin A, two fluorochromes specific for the bases A-T and G-C, respectively. Two models were used for the estimation of the proportion of guanine plus cytosine in the nuclear genome. The first one was based on the linear relationships mithramycin A fluorescence/G-C content and ethidium bromide fluorescence/total DNA content. The second model, based on the curvilinear relationships Hoechst 33342 fluorescence/A-T content and mithramycin A fluorescence/G-C content, resulted in comparatively more homogenous and consistent data and appears more accurate. Comparison with previous reports from other methods for the physical investigation of nuclear genomes shows that flow cytometry of nuclei isolated from protoplasts is an accurate, convenient and robust technique to assay for genome sizes and base pair composition in marine macroalgae.Abbreviations A-T nucleic bases adenine and thymine - CRBC chicken red blood cell - FALS forward-angle light scatter - G-C nucleic bases guanine and cytosine - SEIM sorbitol enzymatic incubation medium - SWIM sea water incubation medium - Tm thermal denaturation temperature of DNA     

Relationships between nuclear DNA content and seed and leaf size in soybean

 A correlation between genome size and agronomically important traits has been observed in many plant species. The goal of the present research was to determine the relationship between genome size, seed size, and leaf width and length in soybean [Glycine max (L.) Merr.] Twelve soybean strains, representing three distinct seed size groups, were analyzed. Flow cytometry was used to estimate their 2C nuclear DNA contents. Data on seed size and leaf size of the 12 strains were obtained from 1994 and 1995 field experiments. Variation of 2C nuclear DNA among the 12 soybean strains was 4.6%, ranging from 2.37 pg for a small-seed strain to 2.48 pg for a large-seed strain. Strain seed size was positively associated with leaf width (r=0.92) and leaf length (r=0.93). Genome size was highly correlated with seed size (r=0.97), leaf width (r=0.90) , and leaf length (r=0.93). The results of our study indicate that there is a significant correlation between genome size and leaf and seed size in soybean. It is possible that selection for greater seed size either leads to, or results from, greater genome size. If so, this relationship might be worth exploring at a more fundamental level. Received: 5 April 1997 / Accepted: 9 January 1998     

Phylogenetic characterisation of picoplanktonic populations with high and low nucleic acid content in the North Atlantic Ocean

In flow cytometric analyses of marine prokaryotic picoplankton often two populations with distinct differences in their apparent nucleic acid content are discernable, one with a high and one with a low nucleic acid content (HNA and LNA, respectively). In this study we determined the phylogenetic composition of flow cytometrically sorted HNA and LNA populations, collected at six stations along a transect across three oceanic provinces from Iceland to the Azores. Catalysed reporter deposition fluorescence in situ hybridisation (CARD-FISH) analysis of sorted cells revealed distinct differences in phylogenetic composition between the LNA and HNA populations with only little overlap. At all stations the LNA population was dominated by the alphaproteobacterial clade SAR11 (45–74%). Also, Betaproteobacteria were always present at 2–4%. While the LNA composition was rather stable, the HNA populations were composed of distinct phylogenetic clades in the different oceanic provinces of Arctic and Tropics. For example Cyanobacteria dominated the North Atlantic Gyre HNA population (29–44%) with Prochlorococcus as the major clade (34–44%), but were low in Arctic and Polar waters (1% and 5%, respectively). In contrast, Bacteroidetes accounted for the majority of HNA cells in the Polar and Arctic province (26% and 32%, respectively), but were low in the Gyre region (3–10%). The DNA content of the HNA population was about 3.5 times higher than that of the LNA populations. This reflects differences in the genome sizes of closely related cultured representatives of HNA clades (3–6 Mbp) and LNA clades (1.3–1.5 Mbp).     

The Determination of Nuclear DNA Content (2C value) on Some Representative Genus and Species of Magnoliaceae

The plant nuclear DNA content (2C value) is a principal characteristic parmeter to describe biodiversity of species,which has important significance on the study of plant resources. In this study, we choosed 23 species from 10 representative genera (Magnolia、Michelia、Manglietia、Liriodendron、Talauma、 Paramichelia、Tsoongiodendron、Manglietiastrum、Kmeria、Parakmeria) of Magnoliaceae in China. All samples were determined using a flow cytometry technique with a standard of Zea mays (545pg/2C). The amount of nuclear DNA among these species ranged from 325pg (317850Mbp) to 1361pg (1331058Mbp) for Pgrandiflora, Mofficinalis subsp. biloba respectively, and the coefficient of variation (CV) were less than 5%. The results of the study will not only provide references for determination of the nuclear DNA content of Magnoliaceae and other plants, but also lay the foundation for the utilization and conservation of Magnoliaceae plant resources.     

Plant DNA flow cytometry and estimation of nuclear genome size

BACKGROUND: DNA flow cytometry describes the use of flow cytometry for estimation of DNA quantity in cell nuclei. The method involves preparation of aqueous suspensions of intact nuclei whose DNA is stained using a DNA fluorochrome. The nuclei are classified according to their relative fluorescence intensity or DNA content. Because the sample preparation and analysis is convenient and rapid, DNA flow cytometry has become a popular method for ploidy screening, detection of mixoploidy and aneuploidy, cell cycle analysis, assessment of the degree of polysomaty, determination of reproductive pathway, and estimation of absolute DNA amount or genome size. While the former applications are relatively straightforward, estimation of absolute DNA amount requires special attention to possible errors in sample preparation and analysis. SCOPE: The article reviews current procedures for estimation of absolute DNA amounts in plants using flow cytometry, with special emphasis on preparation of nuclei suspensions, stoichiometric DNA staining and the use of DNA reference standards. In addition, methodological pitfalls encountered in estimation of intraspecific variation in genome size are discussed as well as problems linked to the use of DNA flow cytometry for fieldwork. CONCLUSIONS: Reliable estimation of absolute DNA amounts in plants using flow cytometry is not a trivial task. Although several well-proven protocols are available and some factors controlling the precision and reproducibility have been identified, several problems persist: (1) the need for fresh tissues complicates the transfer of samples from field to the laboratory and/or their storage; (2) the role of cytosolic compounds interfering with quantitative DNA staining is not well understood; and (3) the use of a set of internationally agreed DNA reference standards still remains an unrealized goal.     

20个赤苍藤种质资源表型性状的遗传多样性分析及综合评价

分析木本蔬菜赤苍藤不同种质资源的表型性状特点,为筛选出叶大、枝条生长旺盛的赤苍藤优良种源奠定基础,该研究以来自越南及中国3省(区)(广西、广东、福建)的20个赤苍藤种源为对象,对12个叶性状及4个枝条性状进行测定及计算,对各性状进行描述统计、方差分析及性状相关性分析,采用主成分分析法计算各种源主成分得分并对各种源进行聚类,对不同赤苍藤种源表型性状进行统计、分类及评价。结果表明:(1)各赤苍藤种源间大多数叶、枝条性状存在显著或极显著差异。种源内各性状变异系数变化幅度不尽相同。种源间各性状变异系数变化幅度依次为叶功能性状(15.42%～70.01%)>枝条性状(20.57%～71.71%)>叶形态性状(3.39%～20.01%); 种源内表型变异更突出。(2)就性状间相关性而言,叶形态性状及叶功能性状指标间多为极显著相关,新枝数量与节间数、新叶数极显著相关,但与叶形态性状的相关关系不明显。(3)16个表型性状共可提取4个主成分,累计贡献率达85.528%。4个主成分分别体现出叶的形态、枝叶萌发生长、叶形状及叶干物质积累及枝条增粗状况。(4)对20个种源进行聚类分析可分为三大类,一类为叶大、生长良好的种源,一类为叶较小、枝条生长旺盛的种源,还有一类种源综合表现不突出。各大类种源中的小类在地理分布上较为接近。(5)福建安溪及福清的种源可选择为大叶种源; 广西大新、上思和桂平的种源可选择为枝条生长旺盛种源。综合来看,中国福建安溪种源综合表现最佳,其次为中国福建福清、中国广东海丰、中国广西南宁、越南茶陵。广西昭平、宜州的两种源综合表现最差,不适宜广西南宁地区引种栽培。部分种源中存在生长表现突出的优良单株,可将此类植株开发为无性系,进行更深入的种源试验。该研究结果为赤苍藤种源表型性状分析及广西南宁引种初步表现提供了科学依据,为筛选、培育高产赤苍藤品种奠定了基础。     

小城镇土地利用变化的生态效应分析

采用系统生态学方法,尤其是应用了Simpson生物多样性指数法,前后相隔10年为时间跨度,对小城镇发展典型地区浙江省绍兴县某镇土地利用变化导致的生态效应进行了初步定量分析与研究.结果表明,由于土地使用功能的改变,导致了以绿色植物为计数的Simpson指数增加近2倍,而以土壤微生物为参照的工业和居住用地的Simpson指数均高于农业用地,反映该小城镇发展过程中生物多样性明显降低,并出现了局部气候的恶化(表现为镇区内年平均气温上升,而年平均相对湿度下降)等不良生态效应.     

The origin, evolution and proposed stabilization of the terms 'genome size' and 'C-value' to describe nuclear DNA contents

BACKGROUND: Perusing the literature on nuclear 'genome size' shows that the term is not stabilized, but applied with different meanings. It is used for the DNA content of the complete chromosome complement (with chromosome number n), for which others use 'C-value', but also for the DNA content of the monoploid chromosome set only (with chromosome number x). Reconsideration of the terminology is required. AIM: Our purpose is to discuss the currently unstable usage of the terms 'genome size' and 'C-value', and to propose a new unified terminology which can describe nuclear DNA contents with ease and without ambiguity. PROPOSALS: We argue that there is a need to maintain the term genome size in a broad sense as a covering term, because it is widely understood, short and phonetically pleasing. Proposals are made for a unified and consensual terminology. In this, 'genome size' should mean the DNA content based on chromosome number x and n, and should be used mainly in a general sense. The necessary distinction of the kinds of genome sizes is made by the adjectives 'monoploid' and the neology 'holoploid'. 'Holoploid genome size' is a shortcut for the DNA content of the whole chromosome complement characteristic for the individual (and by generalization for the population, species, etc.) irrespective of the degree of generative polyploidy, aneuploidies, etc. This term was lacking in the terminology and is for reasons of linguistic consistency indispensable. The abbreviated terms for monoploid and holoploid genome size are, respectively, Cx-value and C-value. Quantitative data on genome size should always indicate the C-level by a numerical prefix, such as 1C, 1Cx, 2C, etc. The proposed conventions cover general fundamental aspects relating to genome size in plants and animals, but do not treat in detail cytogenetic particularities (e.g. haploids, hybrids, etc.) which will need minor extensions of the present scheme in a future paper.     

The effects of nuclear DNA content (C-value) on the quality and utility of AFLP fingerprints

BACKGROUND AND AIMS: Nuclear DNA content (C-value) varies approximately 1000-fold across the angiosperms, and this variation has been reported to have an effect on the quality of AFLP fingerprints. Various methods have been proposed for circumventing the problems associated with small and large genomes. Here we investigate the range of nuclear DNA contents across which the standard AFLP protocol can be used. METHODS: AFLP fingerprinting was conducted on an automated platform using the standard protocol (with 3 + 3 selective bases) in which DNA fragments are visualized as bands. Species with nuclear DNA contents ranging from 1C = 0.2 to 32.35 pg were included, and the total number of bands and the number of polymorphic bands were counted. For the species with the smallest C-value (Bixa orellana) and for one of the species with a large C-value (Damasonium alisma), alternative protocols using 2 + 3 and 3 + 4 selective bases, respectively, were also used. KEY RESULTS: Acceptable AFLP traces were obtained using the standard protocol with 1C-values of 0.30-8.43 pg. Below this range, the quality was improved by using 2 + 3 selective bases. Above this range, the traces were generally characterized by a few strongly amplifying bands and noisy baselines. Damasonium alisma, however, gave more even traces, probably due to it being a tetraploid. CONCLUSIONS: We propose that for known polyploids, genome size is a more useful indicator than the 1C-value in deciding which AFLP protocol to use. Thus, knowledge of ploidy (allowing estimation of genome size) and C-value are both important. For small genomes, the number of interpretable bands can be increased by decreasing the number of selective bases. For larger genomes, increasing the number of bases does not necessarily decrease the number of bands as predicted. The presence of a small number of strongly amplifying bands is likely to be linked to the presence of repetitive DNA sequences in high copy number in taxa with large genomes.     

Ecological effects of cell-level processes: genome size,functional traits and regional abundance of herbaceous plant species

Background and Aims

Genome size is known to be correlated with a number of phenotypic traits associated with cell sizes and cell-division rates. Genome size was therefore used as a proxy for them in order to assess how common plant traits such as height, specific leaf area and seed size/number predict species regional abundance. In this study it is hypothesized that if there is residual correlation between genome size and abundance after these traits are partialled out, there must be additional ecological effects of cell size and/or cell-division rate.

Methods

Variation in genome size, plant traits and regional abundance were examined in 436 herbaceous species of central European flora, and relationships were sought for among these variables by correlation and path analysis.

Key Results

Species regional abundance was weakly but significantly correlated with genome size; the relationship was stronger for annuals (R² = 0·145) than for perennials (R² = 0·027). In annuals, genome size was linked to abundance via its effect on seed size, which constrains seed number and hence population growth rate. In perennials, it weakly affected (via height and specific leaf area) competitive ability. These relationships did not change qualitatively after phylogenetic correction. In both annuals and perennials there was an unresolved effect of genome size on abundance.

Conclusions

The findings indicate that additional predictors of regional abundance should be sought among variables that are linked to cell size and cell-division rate. Signals of these cell-level processes remain identifiable even at the landscape scale, and show deep differences between perennials and annuals. Plant population biology could thus possibly benefit from more systematic use of indicators of cell-level processes.     

Ecological engineering, design, and construction considerations for marsh restorations in Delaware Bay, USA

Public Service Electric & Gas of New Jersey is restoring approximately 4050 ha of salt marsh along Delaware Bay, USA, to offset possible effects on fish populations in the Bay from their existing once-through cooling system. Planning for this effort started with addressing three questions: Do marshes contribute significantly to fish production? How much marsh produces how much fish? Which marshes should be restored? There is ample evidence that salt marshes produce fish. The area of marsh necessary to offset potential losses was calculated from a simple aggregated food chain model and multiplied by four to provide a comfort level to the regulatory agencies. Marshes chosen for restoration were former salt marshes at appropriate tidal elevations. Planning involved experts in marsh ecology, hydrology, and engineering working with the company and regulatory agencies to establish clearly defined goals for the project. Design followed the advice of the experts and construction was overseen to follow the design. Long-term follow up is through adaptive management that is scheduled to continue for about a decade, depending upon progress of the restoration toward its goals.     

Modeling habitat distribution of Cornus officinalis with Maxent modeling and fuzzy logics in China

Aims Predicting suitable habitat distribution is an effective way to protect rare or endangered medicinal plants. Cornus officinalis is a perennial tree growing in forest edge and its air-dried pericarp is one of the traditional Chinese medicines (TCM) with significant medicinal values. In recent years, C. officinalis has undergone severe degeneration of its natural habitat owing to growing market demands and unprecedented damage to the forests. Moreover, the degeneration of suitable habitat has threatened the supply of medicinal materials, and even led to the extinction of some engendered medicinal plant species. In this case, there is a great risk to introduce and cultivate medicinal plants if planners determine the suitable cultivation regions based on personal subjective experience alone. Therefore, predicting suitable potential habitat distribution of medicinal plants (e.g. C. officinalis) and revealing the environmental factors determining such distribution patterns are important to habitat conservation and environmental restoration.Methods In this article, we report the results of a study on the habitat distribution of C. officinalis using maximum entropy (Maxent) modeling and fuzzy logics together with loganin content and environmental variables. The localities of 106 C. officinalis in China were collected by our group and other researchers and used as occurrence data. The loganin content of 234 C. officinalis germplasm resources were tested by high-performance liquid chromatography (HPLC) and used as content data. 79 environmental variables were selected and processed with multicollinearity test by using Pearson Correlation Coefficient (r) to determine a set of independent variables. The chosen variables were then processed in the fuzzy linear model according to the cell values (maximum, minimum) of localities with estimated loganin content. The SDMtoolbox was used to spatially rarefy occurrence data and prepare bias files. Furthermore, combined Maxent modeling and fuzzy logics were used to predict the suitable habitat of C. officinalis. The modeling result was validated using null-model method.Important findings As a result, six environmental factors including tmin3, prec3, bio4, alt, bio12 and bio3 were determined as key influential factors that mostly affected both the habitat suitability and active ingredient of C. officinalis. The highly suitable regions of C. officinalis mainly distribute in a 'core distribution zone' of the east-central China. The statistically significant AUC value indicated that combined Maxent modeling and fuzzy logics could be used to predict the suitable habitat distribution of medicinal plants. Furthermore, our results confirm that ecological factors played critical roles in assessing suitable geographical regions as well as active ingredient of plants, highlighting the need for effective habitat rehabilitation and resource conservation.     

中国稻种资源农艺性状鉴定、编目和繁种入库概况

我国是亚洲栽培稻起源和分化中心之一.稻作历史悠久,稻区分布广泛,类型繁多.经考察和收集,尤其通过国家"七五"、"八五"和"九五"三个五年计划科技攻关协作,编入目录稻种资源75965份,鉴定和繁种67856份入国家种质库保存,获得150余万个性状数据,提供给国家作物种质资源数据库,为今后深入研究和育种利用奠定了丰富的物质基础.本文就我国稻种资源的考察和收集、农艺性状鉴定和编目、繁种与保存、存在问题加以论述,并对今后的稻种资源研究提出一些建议.     

Chromosome identification and karyotype analysis of Podophyllum hexandrum Roxb. ex Kunth using FISH

Podophyllum hexandrum is an important high altitude medicinal plant from Himalaya. Somatic chromosomes of this species were studied to delineate and physical mapping of repetitive rDNA sites to provide landmarks in chromosome identification. The karyotype formula of this species was found to be 6m + 2sm + 2st + 2t with secondary constriction in the chromosome 1 and 7. The FISH analysis of rDNA sites showed 4 sites for 18S rDNA and 2 sites for 5S rDNA. The chromosome number 1, 2, 5 and 6 can be identified based on 18S rDNA sites in their short arm and chromosome 1 and 2 can be identified by 5S rDNA site in the centromere region. The estimated genome size of this plant is 16.07 pg (1C).     

2-苯乙醇对酿酒酵母生理生化特性影响

【目的】研究在不同浓度2-苯乙醇作用下,酵母生理生化特性的变化规律,为优化2-苯乙醇生物合成过程提供重要依据。【方法】透射电镜观察细胞形态;流式细胞术检测细胞膜渗透性、胞内ROS浓度、线粒体膜电位;实时荧光定量PCR检测关键酶基因表达。【结果】随着2-苯乙醇浓度增加(从0到4.0 g/L),酵母细胞分解代谢能力、细胞膜渗透性及aro10基因表达量逐渐降低;线粒体膜电位逐渐增加;胞内ROS浓度先增加后减少。当2-苯乙醇浓度从2.4 g/L增加到3.0 g/L,酵母的分解代谢能力、细胞膜渗透性、aro10基因表达水平等生理生化特性都发生较为显著的变化。【结论】产物原位转移过程中水相2-苯乙醇浓度可考虑控制在2.4 3.0 g/L。     

An exploration of genome size diversity in dragonflies and damselflies (Insecta: Odonata)

Like most insect orders, the Odonata (dragonflies and damselflies) remain poorly studied from the perspective of genome size. They exhibit several characteristics that make them desirable targets for analysis in this area, for example a large range in body size, differences in developmental rate, and distinct modes of flight – all of which are related to genome size in at least some animal taxa. The present study provides new genome size estimates and morphometric data for 100 species of odonates, covering about 1/5 of described North American diversity. Significant relationships are reported between genome size and body size (positive in dragonflies, negative in damselflies), and there is also indication that developmental rate and flight are related to genome size in these insects. Genome size is also positively correlated with chromosome number across the order. These findings contribute to an improved understanding of genome size evolution in insects, and raise several interesting questions for future research.     

The nuclear genome of the phytoseiid <Emphasis Type="Italic">Metaseiulus occidentalis</Emphasis> (Acari: Phytoseiidae) is among the smallest known in arthropods

The genome size of the phytoseiid Metaseiulus (=Typhlodromus or Galendromus) occidentalis (Nesbitt) needs to be estimated before the whole nuclear genome can be sequenced. Two different procedures were used to estimate the genome size of M. occidentalis; (1) flow cytometry (Marescalchi et al. in Genome 33:789–793, 1990) and (2) quantitative real-time PCR (qRT-PCR) (Wilhelm et al. in Nucleic Acids Res 31:e56, 2003). Fluorescence intensity of propidium iodide-stained nuclei of M. occidentalis was measured by flow cytometry using females, males, and eggs. Only the eggs yielded peaks, which ranged in size from 35 to 160 Mb, with a tall peak of 140 Mb in 1-day-old eggs and 65 Mb in 2-day-old eggs, respectively. However, the peaks are broad and do not provide an accurate estimate. The qRT-PCR procedure required single-copy nuclear gene sequences from this phytoseiid. This was accomplished by designing degenerate primers, amplifying the Actin and EF1α sequences from M. occidentalis, and then designing M. occidentalis-specific primers that amplified a unique sequence. The standard qRT-PCR protocol was inefficient and amplification failed frequently, so we developed a high-fidelity qRT-PCR protocol, which utilizes a mix of two DNA polymerases (Taq and a proof-reading Tgo or ACCUZYME) to consistently amplify sequences. This allowed us to estimate the nuclear genome size of M. occidentalis as 88–90 ± 5 Mb. When compared to other arthropod genomes, this appears to be very small.     

Analysis of over-consumption of natural resources and the ecological trade deficit in China based on ecological footprints

China has experienced unprecedented economic development in recent years and is now facing severe challenges caused by the over-consumption of resources and by ecological and environmental degradation. To assess the influence of resource exploitation and ecological trade, we have developed an index of excessive resource consumption based on the concepts of ecological deficit and ecological over-shoot, and we have used the ecological trade deficit to assess the pressure created by the export and import of resources and products. Our analysis indicated that China's consumption footprint surpassed its biocapacity in 1983, leading to an ecological deficit, and the production footprint surpassed its biocapacity in 1986, leading to an ecological over-shoot, as the over-consumption of natural resources grew. By 2010, 3.6 times the current area of bioproductive land was needed to provide sufficient resources to meet the consumption. China has been encouraging the development of exporting enterprises by implementing a series of financial and tax incentives, which have stimulated the economy in the short-term but have gradually increased the ecological trade deficit since 2000.