Pollinator shift and reproductive performance of the qinghai-tibetan plateau endemic and endangered Swertia przewalskii (Gentianaceae)

Reproductive failure results in many plant species becoming endangered. However, little is known of how and to what extent pollinator shifts affect reproductive performance of endangered species as a result of the artificial introduction of alien insects. In this study we examined breeding systems, visitor species, visiting frequency and seed set coefficients of Swertia przewalskii in two years that had different dominant pollinator species (native vs. alien). Flowers of this species were protandrous and herkogamous and insects were needed for the production of seeds. The stigmatic receptivity of this species was shorter than for other gentians. No significant difference in seed set coefficient was found for hand-pollinated plants between the two years, indicating that pollinator shift only had a minor effect on this plant's breeding system. The commonest pollinators in 2002 were native bumblebees, alien honeybees and occasional solitary bees, however, only alien honeybees were observed in 2004. The flower visitation rate in both years was relatively high, although the total visit frequency decreased significantly in 2004. The control flowers without any treatment produced significantly fewer seed sets in 2004 than in 2002. In the past decade the seed production of this species may have partly decreased due to pollination by alien honeybees, however, we suggest that they might have acted as alternative pollinators ensuring seed production of S. przewalskii when native pollinators were unavailable. The main reason that this plant is endangered is probably the result of habitat destruction, but changes in land use, namely intensified agricultural practice and unfavorable animal husbandry have also contributed to its decline. We recommend that in-situ conservation, including the establishment of a protected area, is the best way to preserve this species effectively.

Determination of free fatty acids in soil and bryophyte plants by precolumn derivatization via HPLC with fluorescence detection and tandem mass spectrometry (HPLC-MS/MS)

A sensitive method for the determination of 30 kinds of free fatty acids (FFAs, C-1-C-30) with 1-[2-(p-toluenesulfonate)-ethyl]-2-phenylimidazole-[4,5-f] 9,10-phenan- threne (TSPP) as labeling reagent and using high performance liquid chromatography with fluorescence detection and identification by online postcolumn mass spectrometry with atmospheric pressure chemical ionization (APCI) source in positive-ion mode (HPLC/MS/APCI) has been developed. TSPP could easily and quickly label FFAs in the presence of K2CO3 catalyst at 90 degrees C for 30 min in N,N-dimethylformamide (DMF) solvent, and maximal labeling yields close to 100% were observed with a 5-fold excess of molar reagent. Derivatives were stable enough to be efficiently analyzed by high performance liquid chromatography. TSPP was introduced into fatty acid molecules and effectively augmented MS ionization of fatty acid derivatives and led to regular MS and MS/MS information. The collision induced cleavage of protonated molecular ions formed specific fragment ions at m/z [MH](+)(molecular ion), m/z [M'+CH2CH2](+)(M' was molecular mass of the corresponding FFA) and m/z 295.0 (the, mass of protonated molecular core structure of TSPP). Fatty acid derivatives were separated on a reversed-phase Eclipse XDB-C-8 column (4.6 x 150 mm, 5 mu m, Agilent) with a good baseline resolution in combination with a gradient elution. Linear ranges of 30 FFAs are 2.441 x 10(-3) to 20 mu mol/L, detection limits are 3.24 similar to 36.97 fmol (injection volume 10 mu L, at a signal-to-noise ratio of 3, S/N 3:1). The mean interday precision ranged from 93.4 to 106.2% with the largest mean coefficients of variation (R.S.D.) < 7,5%. The mean intraday precision for all standards was < 6.4% of the expected concentration. Excellent linear responses were observed with correlation coefficients of > 0.9991. Good compositional data could be obtained from the analysis of extracted fatty acids from as little as 200 mg of bryophyte plant samples.Therefore, the facile TSPP derivatization coupled with HPLC/MS/APCI analysis allowed the development of a highly sensitive method for the quantitation of trace levels of short and long chain fatty acids from biological and natural environmental samples.

Determination of long-chain fatty acids in bryophyte plants extracts by HPLC with fluorescence detection and identification with MS

A sensitive method for the determination of long-chain fatty acids (LCFAs) (>C20) using 1-[2-(p-toluenesulfonate)-ethyl]-2-phenylimidazole-[4.5-f]-9,10-phenanthrene (TSPP) as tagging reagent with fluorescence detection and identification with post-column APCI/MS has been developed. The LCFAs in bryophyte plant samples were obtained based on distillation extraction with 1: 1 (v/v) chloroform/methanol as extracting solvent. TSPP could easily and quickly label LCFAs at 90 degrees C in the presence of K2CO3 catalyst in DMF. Eleven free LCFAs from the extracts of bryophyte plants were sensitively determined. Maximal labeling yields close to 100% were observed with a five-fold excess of molar reagent. Separation of the derivatized fatty acids exhibited a good baseline resolution in combination with a gradient elution on a reversed-phase Eclipse XDB-C-8 column. Calculated detection limits from 1.0 pmol injection, at a signal-to-noise ratio of 3, were 26.19-76.67 fmol. Excellent linear responses were observed with coefficients of >0.9996. Good compositional data were obtained from the analysis of the extracted LCFAs containing as little as 0.2 g of bryophyte plant samples. Therefore, the facile TSPP derivatization coupled with HPLC/APCI/MS analysis allowed the development of a highly sensitive method for the quantitation of trace levels of LCFAs from biological and natural environmental samples. (c) 2006 Elsevier B.V. All rights reserved.

Independence of alpine meadow plants from soil organic matter for nitrogen supply: evidence from stable nitrogen isotopes

Stable nitrogen isotope signatures of major sources of mineral nitrogen ( mineralization of soil organic nitrogen, biological N-2 fixation by legumes, annual precipitation and plant litter decomposition) were measured to relatively define their individual contribution to grass assimilation at the Haibei Alpine Meadow Ecosystem, Qinghai, China. The results indicated that delta N-15 values (- 2.40 parts per thousand to 0.97 parts per thousand) of all grasses were much lower than those of soil organic matter (3.4 +/- 0.18 parts per thousand) and mineral nitrogen ( ammonium and nitrate together,7.8 +/- 0.57 parts per thousand). Based on the patterns of stable nitrogen isotopes, soil organic matter (3.4 +/- 0.18 parts per thousand), biological N-2 fixation (0 parts per thousand), and precipitation (- 6.34 +/- 0.24 parts per thousand) only contributed to a small fraction of nitrogen requirements of grasses, but plant litter decomposition (- 1.31 +/- 1.01 parts per thousand) accounted for 67%.

Floral closure in response to temperature and pollination in Gentiana straminea Maxim. (Gentianaceae), an alpine perennial in the Qinghai-Tibetan Plateau

Floral closure may be induced by pollination and various other factors, but is rarely studied comprehensively. Different kinds of floral closure should have various effects on reproductive fitness of plants. Two contrasting types of floral closure were observed in the flowers of Gentiana straminea Maxim. in the eastern Qinghai-Tibetan Plateau. The first type occurred prior to pollination during both gender phases, in response mainly to decreasing air temperatures. Flowers closed when decreasing temperatures approached 20 degrees C and subsequently began to reopen the following day during mid-morning when air temperatures warmed to approximately 13-15 degrees C. This kind of floral closure can protect pollen grains on either stamens or stigmas, increasing fitness of both male and female. Following pollination, permanent floral closure occurred, although there was a delay between the dates of pollination and permanent closure, during which flowers continued to show temporary closure in response to low temperature episodes. The time required for permanent, pollination-induced closure varied according to the age of the gender phase, including a prolonged time before closure if pollination occurred early in the female phase. The retaining of permanent closed flowers increased both approaching (to inflorescences) and visiting (to unpollinated flowers) frequencies of individual plants when with fewer open flowers and the persisting corolla is further beneficial for seed sets of these pollinated flowers. Thus, two separate types of floral closure, one in response to environmental cues and the other in response to the age of each gender stage, appeared to have a strong influence on reproductive fitness in this species. These results revealed a different adaptive strategy of alpine plants in the sexual reproduction assurance in addition to the well-known elevated floral longevity, dominant role of more effective pollinators and increased reproduction allocation in the arid habitats.

Development of a sensitive fluorescent derivatization reagent 1,2-benzo-3,4-dihydrocarbazole-9-ethyl chloroformate (BCEOC) and its application for determination of amino acids from seeds and bryophyte plants using high-performance liquid chromatography with fluorescence detection and identification with electrospray ionization mass spectrometry

A pre-column derivatization method for the sensitive determination of amino acids and peptides using the tagging reagent 1,2-benzo-3,4dihydrocarbazole-9-ethyl chloroformate (BCEOC) followed by high-performance liquid chromatography with fluorescence detection has been developed. Identification of derivatives was carried out by liquid chromatography/electrospray ionization mass spectrometry (LC/ESI-MS/MS). The chromophore of 2-(9-carbazole)-ethyl chloroformate (CEOC) reagent was replaced by 1,2-benzo-3,4-dihydrocarbazole functional group, which resulted in a sensitive fluorescence derivatizing reagent BCEOC. BCEOC can easily and quickly label peptides and amino acids. Derivatives are stable enough to be efficiently analyzed by high-performance liquid chromatography. The derivatives showed an intense protonated molecular ion corresponding m/z (M + H)(+) under electrospray ionization (ESI) positive-ion mode with an exception being Tyr detected at negative mode. The collision-induced dissociation of protonated molecular ion formed a product at m/z 246.2 corresponding to the cleavage of C-O bond of BCEOC molecule. Studies on derivatization demonstrate excellent derivative yields over the pH 9.0-10.0. Maximal yields close to 100% are observed with a 3-4-fold molar reagent excess. Derivatives exhibit strong fluorescence and extracted detzvatization solution with n-hexane/ethyl acetate (10:1, v/v) allows for the direct injection with no significant interference from the major fluorescent reagent degradation by-products, such as 1,2-benzo-3,4-dihydrocarbazole-9-ethanol (BDC-OH) (a major by-product), mono- 1,2-benzo-3,4-dihydrocarbazole-9-ethyl carbonate (BCEOC-OH) and bis-(1,2-benzo-3,4-dihydrocarbazole-9-ethyl) carbonate (BCEOC)(2). In addition, the detection responses for BCEOC derivatives are compared to those obtained with previously synthesized 2-(9-carbazole)-ethyl chloroformate (CEOC) in our laboratory. The ratios AC(BCEOC)/AC(CEOC) = 2.05-6.51 for fluorescence responses are observed (here, AC is relative fluorescence response). Separation of the derivatized peptides and amino acids had been optimized on Hypersil BDS C-18 column. Detection limits were calculated from 1.0 pmol injection at a signal-to-noise ratio of 3, and were 6.3 (Lys)-177.6 (His) fmol. The mean interday accuracy ranged from 92 to 106% for fluorescence detection with mean %CV < 7.5. The mean interday precision for all standards was < 10% of the expected concentration. Excellent linear responses were observed with coefficients of > 0.9999. Good compositional data could be obtained from the analysis of derivatized protein hydrolysates containing as little as 50.5 ng of sample. Therefore, the facile BCEOC derivatization coupled with mass spectrometry allowed the development of a highly sensitive and specific method for the quantitative analysis of trace levels of amino acids and peptides from biological and natural environmental samples. (c) 2005 Elsevier B.V. All rights reserved.

The effect of land management on carbon and nitrogen status in plants and soils of alpine meadows on the Tibetan plateau

Large-scale grassland rehabilitation has been carried out on the severely degraded lands of the Tibetan plateau. The grasslands created provide a useful model for evaluating the recovery of ecosystem properties. The purposes of this research were: (1) to examine the relative influence of various rehabilitation practices on carbon and nitrogen in plants and soils in early secondary succession; and (2) to evaluate the degree to which severely degraded grassland altered plant and soil properties relative to the non-disturbed native community. The results showed: (1) The aboveground tissue C and N content in the control were 105-97 g m(-2) and 3.356gm(-2), respectively. The aboveground tissue C content in the mixed seed treatment, the single seed treatment, the natural recovery treatment and the severely degraded treatment was 137 per cent, 98 per cent, 49 per cent and 38 per cent, respectively, of that in the control. The corresponding aboveground tissue N content was 109 per cent, 84 per cent, 60 per cent and 47 per cent, respectively, of that in the control. (2) Root C and N content in 0-20 cm depths of the control had an 2 2 average 1606 gm(-2) and 30-36 gm(-2) respectively. Root C and N content in the rehabilitation treatments were in the range of 26-36 per cent and 35-53 per cent, while those in the severely degraded treatment were only 17 per cent and 26 per cent of that in the control. (3) In the control the average soil C and N content at 0-20 cm was 11307 gm(-2) and 846 gm(-2), respectively. Soil C content in the uppermost 20 cm in the seeded treatments, the natural recovery treatment and the severely degraded treatment was 67 per cent, 73 per cent and 57 per cent, respectively, while soil N content in the uppermost 20cm was 72 per cent, 82 per cent and 79 per cent, respectively, of that in the control. The severely degraded land was a major C source. Restoring the severely degraded lands to perennial vegetation was an alternative approach to sequestering C in former degraded systems. N was a limiting factor in seeding grassland. It is necessary for sustainable utilization of seeding grassland to supply extra N fertilizer to the soil or to add legume species into the seed mix. Copyright (c) 2005 John Wiley & Sons, Ltd.

陆生蜗牛壳体同位素组成及其对生长季节的响应

Stable isotope compositions of land snail shells have a great potential as an indicator of paleoclimatic and paleoenvironmental changes. However, some key issues, such as the relationship of carbon isotope between snail food and local vegetation, and the uncertainty of the dominant factors about snail body fluid changes in oxygen isotope composition, remain less well known, strongly limiting shell isotopic application. In this study, we measure the stable isotope compositions on the shells of both live snails and fossils collected from the Chinese Loess Plateau and a loess sequence at Mangshan, Xingyang, respectively. Based on the analyses, the association of the stable isotope compositions of land snail shells with their growing seasons is investigated. In addition, the climatic and environmental significances of isotopic differences among several snail species are discussed. The main results and conclusions are presented as follows: 1. δ18O values for the shell lip samples of Bradybaena ravida redfieldi range from -6.79‰ to -1.92‰, and parallels to the monthly changes of local rain water δ18O, temperature and humidity. The compatibility of shell lip δ18O with monthly modeled shell δ18O indicates that the shell lip δ18O changes are mainly resulted from the 18O variations of rain-water. The shells of a land snail growing in spring could be enriched in 18O, and those growing in summer depleted in 18O. 2. Carbon isotope compositions of snail shells are controlled by their diet, which is affected by the relative proportion of C3 to C4. There are some differences in carbon isotopic compositions among different snail species, especially between P. orphana and V. tenera or P. aeoli. Shell δ13C for P. orphana is the most positive with an average of -5.88 ± 2.54 ‰. The C4 plant fraction of the food for “cold-aridiphilous” taxa, P. aeoli and V. tenera, is distinctly lower than that for “thermo-humidiphilous” taxa, P. orphana, indicating that summer is likely to be the main active season of P. orphana and spring of P. aeoli and V. tenera. Therefore, some discrepancy of carbon isotopic compositions among different species may be related to snail active season. 3. δ13C values among different species have a certain degree of positive correlation, which may be influenced by local vegetation ecosystem. δ13C value of the snail shells (especially P. orphana) shows an eastward increasing trend and consists with the variations of C4 plants biomass in Loess Plateau. The result shows that the carbon isotope in local vegetation ecosystem is one of the main factors influencing δ13C values of snail food. Therefore, both carbon isotopes of local vegetation ecosystem and snail active season contribute to the carbon isotopic differences among different snail species and in different areas. 4. δ13C values of living snail shells and soil organic matter have a positive correlation with each other, which further supports the view that carbon isotope in local vegetation ecosystem is one of the main factors influencing δ13C values of snail food. However, the range of δ13C values of snail food for various species in response to carbon isotope in local vegetation ecosystem is different. It is suggested that 13C enrichment of snail shells relative to local vegetation ecosystem has a potential to indicate snail active season and the degree of climate temperature and humidity. 5. There is a significant negative correlation between carbon and oxygen isotopic compositions of living snail shells in Loess Plateau. This result further supports that snail active season can be inferred based on the shell carbon and oxygen isotopic compositions. Moreover, there are some positive correlations between mean annual temperature and differences of shell δ13C values ( 13CV. tenera-P. orphana) and that of δ18O values ( 18OV. tenera-P. orphana) for P. orphana, a typical “thermo-humidiphilous” taxa, and V. tenera, a typical “cold-aridiphilous” taxa, respectively. It shows that  13CV. tenera-P. orphana and  18OV. tenera-P. orphana may have a potential to indicate mean annual temperature or the length of biological growing season. 6. Stable isotopes of land snail shell in the Mangshan loess sequence show that the shell δ18O value of “cold-aridiphilous” taxa V. tenera is more positive than “thermo-humidiphilous” taxa P. orphana and δ13C value of the former is more negative than the latter. In addition, the shell δ18O value of V. tenera varies significantly in different period. During the last glacial maximum, its δ18O value with an average of -7.89 ‰ is more negative than that (-5.88 ‰) from the last deglaciation to the early Holocene. This phenomenon indicates that its growing season during different period is significantly different. It tends to grow in summer in last glacial maximum. With climate warming, it prefers growing in spring with relatively low temperature. While the shell δ18O value of P. orphana varies in a little range, which shows that its activity season is shorter and mainly in summer. These results further support that the change of the snail growing season is one of the main factors of differences of carbon isotopic compositions among different snail species and varies with time. Furthermore, it is consistent that changes in magnetic susceptibility and trend of differences of shell δ18O values and δ13C values respectively between the two snail fossils. It is further testified that 13CV. tenera-P. orphana and  18OV. tenera-P. orphana may have a potential to indicate mean annual temperature or the length of biological growing season.

中国北方草本植物及表土有机质碳同位素组成

Recently, more and more attention has been paid to stable isotope ratios in terrestrial depositional systems. Among them, δ~(13)C value is mainly determined by the surface vegetation, while vegetation is directly related to climate, therefore, carbon isotope ratio in soil organic matter and pedogenic carbonate has been employed as an important paleoecological indicator. In order to test the paleoecological information extracted from stable isotope ratios in terrestrial depositional systems, it is necessary to study the relationships between δ~(13)C value in standing terrestrial plants and today climate, as well as between δ~(13)C value in modern surface soil organic matter and standing vegetation. Thus, these relationships were studied in this paper by means of analysing δ~(13)C in standing plants and modem surface soil organic matter in North China. The main results and conclusions are presented as following: 1. According to their δ~(13)C values, 40 C-4 species represent about 16% of the 257 plant species sarnpled from the North China. C-4 photosynthesis mainly occurs in Poaceae, Cyperaceae and Chenopidaceae families, and percentage representation of C-4 photosynthesis is up to 56% in Poaceae family. 2. The δ~(13)C values of C-3 plant species in North China vary from -21.7‰ to -32.0‰ with an average of -27.1‰, and 93% focus on the range of -24.0‰ ～ -30.0‰; δ~(13)C values of C-4 plant species in North China are between -10.0‰ ～ -15.5‰ with an average of -12.9‰, and 90% concentrate on the range of -11.0‰ ～ -15.0‰. 3. The δ~(13)C composition of C-3 plant species collected from Beijing, a semi-moist district, mainly vary between -27.0‰ ～ -30.0‰, and the average is -28.7‰; the δ ~(13)C values of plants in the semi-arid district, east and west to the Liu Pan Moutain, focus on the range of-26.0‰ ～ -29.0‰ and -25.0‰ ～ -28.0‰, respectively, with the mean value of -27.6‰ and -26.6‰, respectively; the δ~(13)C composition in the arid district dominantly vary from -24.0‰ to -29.0‰, with the average of -26.2‰, and among them, the δ~(13)C values of C-3 plant species in deserts are often between - 22‰ ～ -24‰; the δ~(13)C values in the cold mountain district concentrate on the range of -24.0‰ to -29.0‰, with the average of -26.3‰. 4. The main range of δ~(13)C composition of C-4 plant species, derived from Beijing, a semi-moist district, are -13.0‰ ～ -15.5‰; the semi-arid district, -11.0‰ ～ -14.0‰; the arid district, -11.0‰ ～ -14.0‰. The mean values of them are -14.0‰, -12.4‰,-12.7‰, respectively. 5. From east to west in North China, δ~(13)C values of C-3 plant species increase with longitude. The correlation between δ~(13)C ratios of C-3 plant species and longitude is linear. Changing temperate and precipitation and changing atmosphere pressure are spossible explanations. 6. Almost all C-3 plant species have the trends that their δ~(13)C values gradually increase with decreasing precipitation, decreasing temperature and increasing altitude. Our results show the increases of the δ~(13)C value by 0.30 ～ 0.45‰, 0.19 ～ 0.27‰ and 1.1 ～ 1.2‰ per 100 mm, I℃ and 1000 m, respectively, for all C-3 plant species together. 7. The δ~(13)C values of all C-3 plant species together and a part of C-3 species show highly significant linear correlation with the mean annual temperature, the mean annual precipitation and the altitude, and the results suggest that they can be used as proxies of these environmental variables, while, those without highly significant correlation, may be not suitable as the proxies. 8. The extent, which of responses of δ~(13)C composition to environmental variables, is different for each C-3 plant specie. 9. The δ~(13)C variations along altitude and longitude may be non-linear for C-4 p1ant species in North China. The mean annual temperature may be not important influential factor, thus, it suggests that the δ~(13)C composition of C-4 plant species may be not suitable as the proxy of the mean annual temperature. The influences of summer temperature on δ~(13)C values are much bigger than that of annual temperature, among them, the influence of September temperature is biggest. The mean annual precipitation may be one of the dominant influential factors, and it shows a highly significant non-linear relationship with δ~(13)C values, and the result indicates that δ~(13) C composition of C-4 plant species can be employed as the proxy of the mean annual precipitation. 10. The variations of δ~(13)C ratios do not show systematic trends along longitude, latitude and altitude for modern surface soil organic in Northwest China. ll. The δ~(13)C ratios of modern surface soil organic do not exhibit systematic patterns with temperature and precipitation in Northwest China, it suggests that, unless soil organic is transferred from pure C-3 or C-4 vegetation, the δ~(13)C composition of soil organic may be not used as proxies of climatic variables. 12. The δ~(13)C values of modem surface soil organic are heavier than that of standing vegetation, and the difference ofrnean δ~(13)C between them is -2.18‰. 13. Without considering the δ~(13)C difference between vegetation and soil organic, as well as the δ~(13)C drift in various enviromnent, we may not obtain the valuable information of C-3, C-4 relative biomass in vegetation. 14. The C-4 biomass contribution in vegetation increase with decreasing latitude, increasing longitude and decreasing altitude in Northwest China. The C-4 biomass almost are zero in those regions north to 38 ° N, or west to 100°E, or above 2400 m. 15. The C-4 relative biomass in vegetation increase with growing temperature and precipitation. and, C-4 plants are rare at those regions where the mean annual temperature is less 4 ℃, or the mean annual precipitation is less 200 mm, and their biomass contribution in vegetation are almost zero. Both the mean annual temperature and the mean annual average precipitation may be the important influential factors of C-4 distribution, but the dominant factors.

中国北方典型生态区表土孢粉组合特征及其与现代植被和气候的关系

Surface pollen assemblages and their relationhips with the modern vegetation and climate provide a foundation for investigating palaeo-environment conditions by fossil pollen analysis. A promising trend of palynology is to link pollen data more closely with ecology. In this study, I summarized the characteristics of surface pollen assemblages and their quantitative relation with the vegetation and climate of the typical ecological regions in northern China, based on surface pollen analysis of 205 sites and investigating of modern vegetation and climate. The primary conclusions are as follows:The differences in surface pollen assemblages for different vegetation regions are obvious. In the forest communities, the arboreal pollen percentages are more than 30%, herbs less than 50% and shrubs less than 10%; total pollen concentrations are more than 106 grains/g. In the steppe communities, arboreal pollen percentages are generally less than 5%; herb pollen percentages are more than 90%, and Artemisia and Chenopodiaceae are dominant in the pollen assemblages; total pollen concentrations range from 103 to 106 grains/g. In the desert communities, arboreal pollen percentages are less than 5%. Although Chenopodiaceae and Artemisia still dominate the pollen assemblages, Ephedra, Tamaricaceae and Nitraria are also significant important in the pollen assemblages; total pollen concentrations are mostly less than 104grains/g. In the sub-alpine or high and cold meadow communities, arboreal pollen percentages are less than 30%. and Cyperaceae is one of the most significant-taxa in the pollen assemblages. In the shrub communities, the pollen assemblages are consistent with the zonal vegetation; shrub pollen percentages are mostly less than 20%, except for Artemisia and Hippophae rhamnoides communities.There are obvious trends for the pollen percentage ratios of Artemisia to Chenopodiaceae (A/C), Pinus to Artemisia (P/A) and arbor to non-arbor (AP/NAP) in the different ecological regions. In the temperate deciduous broad-leaved forest region, the P/A ratios are generally higher than 0.1, the A/C ratios higher than 2 and the AP/NAP ratios higher than 0.3. In the temperate steppe regions, the P/A ratios are generally less than 0.1, the A/C ratios higher than 1 and the AP/NAP ratios less than 0.1. In the temperate desert regions, the P/A ratios are generally less than 0.1, the A/C ratios less than 1, and the AP/NAP ratios less than 0.1.The study on the representation and indication of pollen to vegetation shows that Pinus, Artemisia, Betula, Chenopodiaceae, Ephedra, Selaginella sinensis etc. are over-representative in the pollen assemblages and can only indicate the regional vegetation. Some pollen types, such as Quercus, Carpinus, Picea, Abies, Elaeagus, Larix, Salix, Pterocelis, Juglans, Ulmus, Gleditsia, Cotinus, Oleaceae, Spiraea, Corylus, Ostryopsis, Vites, Tetraena, Caragana, Tamaricaceae, Zygophyllum, Nitraria, Cyperaceae, Sanguisorba etc. are under-representative in the pollen assemblages, and can indicate the plant communities well. Populus, Rosaceae, Saxifranaceae, Gramineae, Leguminosae, Compositae, Caprifoliaceae etc. can not be used as significant indicators to the plants.The study on the relation of pollen percentages with plant covers shows that Pinus pollen percentages are more than 30% where pine trees exist in the surrounding region. The Picea+Abies pollen percentages are higher than 20% where the Picea+Abies trees are dominant in the communities, but less than 5% where the parent plants are sparse or absent. Larix pollen percentages vary from 5% to 20% where the Larix trees are dominant in the communities, but less than 5% where the parent plants are sparse or absent. Betula pollen percentages are higher than 40% where the Betula trees are dominant in the communities" but less than 5% where the parent plants are sparse or absent. Quercus pollen percentages are higher than 10% where the Quercus trees are dominant in the communities, but less than 1% where the parent plants sparse or absent. Carpinus pollen percentages vary from 5% to 15% where the Carpinus trees are dominant in the communities, but less than 1% where the parent plants are sparse or absent. Populus pollen percentages are about 0-5% at pure Populus communities, but cannot be recorded easily where the Populus plants mixed with other trees in the communities. Juglans pollen accounts for 25% to 35% in the forest of Juglans mandshurica, but less than 1% where the parent plants are sparse or absent. Pterocelis pollen percentages are less than 15% where the Pterocelis trees are dominant in the communities, but cannot be recorded easily where the parent plants are sparse or absent. Ulmus pollen percentages are more than 8% at Ulmus communities, but less than 1% where the Ulmus plants mixed with other trees in the communities. Vitex pollen percentages increase along with increasing of parent plant covers, but the maximum values are less than 10 %. Caragana pollen percentages are less than 20 % where the Caragana plant are dominant in the communities, and cannot be recorded easily where the parent plants are sparse or absent. Spiraea pollen percentages are less than 16 % where the Spiraea plant are dominant in the communities, and cannot be recorded easily where the parent plants are sparse or absent.The study on the relation of surface pollen assemblages with the modern climate shows that, in the axis 1 of DCA, surface samples scores have significant correlation with the average annual precipitations, and the highest determination coefficient (R2) is 0.8 for the fitting result of the third degree polynomial functions. In the axis 2 of DCA, the samples scores have significant correlation with the average annual temperatures, average July temperatures and average January temperatures, and the determination coefficient falls in 0.13-0.29 for the fitting result of the third degree polynomial functions with the highest determination coefficient for the average July temperature.The sensitivity of the different pollen taxa to climate change shows that some pollen taxa such as Pinus, Quercus, Carpinus, Juglans, Spiraea, Oleaceae, Gramineae, Tamariaceae and Ephedra are only sensitive to the change in precipitation.