Summer Warming Limited Bud Output Drives a Decline in Daughter Shoot Biomass through Reduced Photosynthetis of Parent Shoots in Leymus chinensis Seedlings

Understanding how summer warming influences the parent and daughter shoot production in a perennial clonal grass is vital for comprehending the response of grassland productivity to global warming.Here,we conducted a simulated experiment using potted Leymus chinensis,to study the relationship between the photosynthetic activ-ity of parent shoots and the production of daughter shoots under a whole(90 days)summer warming scenario(+3°C).The results showed that the biomass of parents and buds decreased by 25.52%and 33.45%,respectively,under warming conditions.The reduction in parent shoot biomass due to warming directly resulted from decreased leaf area(18.03%),chlorophyll a(18.27%),chlorophyll b(29.21%)content,as well as a reduction in net photosynthetic rate(7.32%)and the maximum quantum efficiency of photosystem II(PSII)photochemistry(4.29%).The decline in daughter shoot biomass was linked to a decrease in daughter shoot number(33.33%)by warming.However,the number of belowground buds increased by 46.43%.The results indicated that long-term summer warming reduces biomass accumulation in parent shoot by increasing both limitation of stoma and non-stoma.Consequently,the parent shoot allocates relatively more biomass to the belowground organs to maintain the survival and growth of buds.Overall,buds,as a potential aboveground population,could remedy for the cur-rent loss of parent shoot density by increasing the number of future daughter shoots if summer warming subsides.

AFLP Variation Analysis on the Germplasm Resources of Leymus chinensis

The fingerprints of 27 Leymus chinensis (Trin.) Tzvel accessions from different regions in China were generated and their genomic DNA variations were assayed via amplified fragment length polymorphism (AFLP). A total of 537 DNA fragments were amplified ranging from 75-530 bp using 8 AFLP primer combinations, among which 89 (16.6%) were monomorphic and 329 (61.3%) were polymorphic. The average number of DNA band produced by each primer combination was 66.13 and the total polymorphic rate was 78.84%. PICs (polymorphic information contents) scores for AFLP ranged from 0.0-0.5 and the mean PIC was 0.216. PIC scores were maximum (0.5) for 8.5% of the AFLP markers. All the results revealed that there was rich polymorphism in genomic DNA of L. chinensis . A DNA molecular dendrogram was established for 27 L. chinensis genotypes based on genetic variation and UPGMA cluster analysis of 537 AFLP markers amplified by 8 primer combination, suggesting that the genetic variation with high frequency showed close correlation with their ecotopes and geographical distributions. Twenty_seven L. chinensis genotypes could be divided into four groups. Among them the lower genetic relationship was shown. The reason of high genetic variation frequency in L. chinensis species and formation of varieties was discussed.

Variation of Water-Use Efficiency of Leymus chinensis and Cleistogenes squarrosa in Different Plant Communities in Xilin River Basin, Nei Mongol

Water is usually considered to be a key limiting factor for the growth and reproduction of steppe plants in the Xilin River Basin, Nei Mongol. Foliar delta C-13 values, an indicator of long-term intercellular carbon dioxide concentration and thus of water-use efficiency (WUE) in plants, were measured on Leymus chinensis (Trin.) Tzvel. and Cleistogenes squarrosa (Trin.) Keng. in six communities of different habitats in die Xilin River Basin. The foliar delta C-13 values of both species tended to increase with decreasing soil water content (SWC) and a significant negative correlation was found between foliar delta C-13 Values and SWC in different soil layers, indicating that the two species could change WUE according to water availability. We also found relatively constant leaf water contents (LWC) of the two species in different habitats. Our results implied that the two steppe species might have adapted to different soil water regimes either through adjusting stomatal conductance to get a proper WUE, or through enhancing the osmosis-regulating ability to keep a relatively stable LWC. Our findings could partially explain why the two plant species have a wide distribution range and become dominant in the Xilin River Basin.

Water Use of Leymus chinensis Community

Soil moisture of Leymus chinensis (Trin.) Tzvel. community has obviously stratified phenomena: the layer (0-40 cm) in which roots are concentrically distributed is directly influenced by precipitation and evapotranspiration. It can be called interaction layer of precipitation and evapotranspiration. The layer (40-120 cm), where water-storage capacity exchange lagged exchange of the root-layer water-storage capacity and the community evapotranspiration, can be called major water-storage layer. The layer (under 120 cm) can be called water relatively stable/balanced layer. The year 1996 was a normal flow year, and soil water had a surplus of 18 mm at the end of the growing season. The year 1998 was a high flow year, because leakage took place under continuous heavy rainfall, soil water had a deficit of 15 mm at the end of the growing season. Transpiration to evapotranspiration ( T/ET) value reflected not only the luxuriance degree of the community, but also the water use regime of the environmental resources. T/ET value was low (0.5) in May 1998, reaching 0.7 in June, then decreasing to 0.6 in July, due to the impact of rainfall inclining, while August reached the maximum (0.9), and September decreased to 0.6. Water use efficiency (WUE) was mainly restricted by the growing rate of plants under sufficient water condition (1998). Its seasonal changes were coincident with the grand period of growth of the plants. When both meanings of WUE and T/ET were analyzed profoundly, the concept of evapotranspiration efficiency (ETE) which can all-side reflect utilization regime of the environmental water resources was advanced.

Molecular Cloning and Sequence Analysis of Class Ⅱ Chitinase Gene in Leymus chinensis

[ Objective] The aim of this study was to clone Class Ⅱ chitinase gene in Leymus chinensis grown in saline land in Heilongjiang Province and analyze its sequence, which provided a foundation for further study on the biological function and application of chitinasa gene. [ Method] cDNA library of Leymus chinensis leaves were constructed, and its DNA sequence was determined or analyzed, while the homology of chitinasa gene and amino acid sequence was compared with that in GenBank. [ Result] One full length cDNA fragment with length of 996 bp was cloned from cDNA library of Leymus chinensis leaves. The length of ORF was 768 bp encoding 225 amino acids （GenBank accession number： EU344908）. The encoding products lacked CBD and C-terminal extension region from the view of structure, but had structural characters of Class Ⅱ chitinase gene, which indicated that amino acid sequence had high homology compared with Class Ⅱ chitinase gene of rye and wheat. The constructed recombinant vector pQE-LcChi2 could express a protein of 27 kD through induction, which was consistent with the deduced encoding product of pQE-LcChi2 gene. [ Conclusion] LcChl2 gene is an expression gene, which can express in E. coll.

Research on Phenotypic Diversity of Different Populations of Leymus Chinensis in Zhalong Wetlands

[Objective] This paper aimed at revealing the rules of phenotypic variation of Leymus chinensis in Zhalong wetlands and providing the scientific and theoretical bases for devising protection strategies of Leymus chinensis populations in Zhalong National Nature Reserve.[Method] Four phenotypic traits of five Leymus chinensis populations in Zhalong Wetlands were comparison and analysis using principal component analysis method and UPGMA cluster analysis method.[Result] There were significant differences among the four phenotypic traits of five populations,the order of coefficients of variation were node number〉plant height〉leaf width〉leaf length;the order of variation degrees of the populations from large to small was KQH,MD,JZ,ZK,TTG population.Results of principal component analysis suggested that the four traits were all factors led to the phenotypic differences among the populations of Leymus chinensis.Leymus chinensis of five populations were divided into three groups by cluster analysis,MD,JZ and TTG were classified as one group,KQH and ZK population were individually classified as one group.[Conclusion] Leymus chinensis had large-scale phenotypic variations and showed strong adaptability to different habitats,which was very important for selection,breeding of fine varieties and germplasm conservation.

Diurnal and Seasonal Dynamics of Soil Respiration at Temperate Leymus Chinensis Meadow Steppes in Western Songnen Plain, China

To evaluate the diurnal and seasonal variations in soil respiration （Rs） and understand the controlling factors, we measured carbon dioxide （CO2） fluxes and their environmental variables using a LI-6400 soil CO2 flux system at a temperate Leymus chinensis meadow steppe in the western Songnen Plain of China in the growing season （May-October） in 2011 and 2012. The diurnal patterns of soil respiration could be expressed as single peak curves, reaching to the maximum at 11：00-15：00 and falling to the minimum at 21：00-23：00 （or before dawn）. The time-window between 7：00 and 9：00 could be used as the optimal measuring time to represent the daily mean soil CO2 efflux. In the growing season, the daily value of soil CO2 efflux was moderate in late spring （1.06-2.51μnol/（m2.s） in May）, increased sharply and presented a peak in summer （2.95-3.94 μmol/（m2.s） in July）, and then decreased in autumn （0.74-0.97 μmol/（m2.s） in October）. Soil temperature （Ts） exerted dominant control on the diurnal and seasonal variations of soil respiration. The temperature sensitivity of soil respiration （Q10） exhibited a large seasonal variation, ranging from 1.35 to 3.32, and decreased with an increasing soil temperature. Rs gradually increased with increasing soil water content （Ws） and tended to decrease when Ws exceeded the optimum water content （27%） of Rs. The Ts and Ws had a confounding effect on Rs, and the two-variable equations could account for 72% of the variation in soil respiration （p 〈 0.01）.

Linking nutrient strategies with plant size along a grazing gradient： Evidence from Leymus chinensis in a natural pasture

St udying the changes in nutrient use strategies induced by grazing can provide insight into the process of grassland degradation and is important for improving grassland quality and enhancing ecosystem function. Dominant species in meadow steppe can optimize their use of limiting resources; however, the regulation of nutrient use strategies across grazing gradients is not fully understood. Therefore, in this study, we report an in situ study in which the impact of grazing rates on nutrient use strategies of Leymus chinensis, the dominant plant species in eastern Eurasian temperate steppes, was investigated. We conducted a large randomized controlled experiment（conducted continuously for five years in grassland plots in a natural pasture in Ha ilar, eastern Mongolia Plateau, China） to assess the effects of grazing rate treatments（0.00, 0.23, 0.34, 0.46, 0.69, and 0.92 adult cattle unit（AU） ha-1） on L. chinensis along a grazing gradient and employed a random sampling approach to compare the accumulation, allocation, and stoichiometry of C, N, and P in leaves and stems. Ou r findings demonstrated the follows：（i） The height of L. chinensis decreased with an increase in the grazing gradient, and the concentrations of C, N, and P significantly increased;（ii） the accumulation of C, N, and P per individual was negatively correlated with the concentration of aboveground tissues, suggesting that there was a tradeoff in L. chinensis between nutrient accumulation and concentration at the individual scale;（iii） the leaf-to-stem ratio of C, N, and P accumulation increased with grazing intensity, indicating a tradeoff in nutrient allocation and plant size at the individual plant level; and（iv） grazing rates were negatively correlated with the ratios of C：N and C：P in the stem; however, these ratios in leaves significantly increased with grazing intensity. Our findings suggest that L. chinensis in meadow steppe adapts to grazing disturbance through tradeoffs between plant size and nutrient use strategies. Moreover, our results imply that grazing produces a compensatory effect on nutrient use efficiency between the stems and leaves of L. chinensis.

Osmotic Regulation of Betaine Content in Leymus chinensis Under Saline-alkali Stress and Cloning and Expression of Betaine Aldehyde Dehydrogenase(BADH) Gene

The potted Leymus chinensis seedlings were treated with saline-alkali solution of six different（from Ⅰ to Ⅵ） concentrations. The results demonstrate that the betaine content and Betaine-aldehyde dehydrogenase（BADH： EC 1.2.1.8） activities have a direct relation with increased stressing time in the same treatment; both exhibit a single peak with increasing the concentration of saline-alkali solution, and number V shows the highest value. The BADH gene of Leyrnus chinensis was cloned by RT-PCR and RACE technology and was designated as LcBADH. The cDNA sequence of LcBADH was 1774bp including the open reading frame（ORF） of 1521bp（coding 506 amino acids）. The vector of prokaryotic expression was constructed by inserting the LcBADH gene fragmcnt into pET30a（＋） and transformed into E. coli BL21（DE3）. The result of SDS-PAGE shows that the idio-protein with a molecular mass of 56.78 kDa was effectively expressed in the recombinant bacteria induced by isopropyl fl-D-thiogalactoside（IPTG）.

Long-term effects of mowing on plasticity and allometry of Leymus chinensis in a temperate semi-arid grassland,China

Mowing is an important land management practice for natural semi-arid regions. A growing body of empirical evidence shows that different mowing regimes affect the functioning of grassland ecosystems. However, the responses of plant functional traits to long-term mowing and their allometric scaling under long-term mowing are poorly understood. For a better understanding of the effects of mowing on grassland ecosystems, we analyzed the allometric traits of leaves and stems of Leymus chinensis （Trin.） Tzvel., a dominant grass species in eastern Eurasian temperate grassland, at different mowing intensities （no clipping, clipping once every two years, once a year and twice a year）. Experiments were conducted on plots established over a decade ago in a typical steppe of Xilinhot, Inner Mongolia, China. Results showed that most of the functional traits of L. chinensis decreased with the increased mowing intensity. The responses of leaves and stems to long-term mowing were asymmetric, in which leaf traits were more stable than stem traits. Also significant allometric relationships were found among most of the plant functional traits under the four mowing treatments. Sensitive traits of L. chinensis （e.g. leaf length and stem length） were primary indicators associated with aboveground biomass decline under high mowing intensity. In conclusion, the allometric growth of different functional traits of L. chinensis varies with different long-term mowing practices, which is likely to be a strategy used by the plant to adapt to the mowing disturbances.

Effect of Nitrogen Fertilizer on Photosynthetic Rate of Leymus chinensis in Grassland of Different Degrading Degrees

[Objective] This study aimed to investigate the effect of nitrogen fertilizer on photosynthetic rate of Leymus chinensis in the grasslands of different degrading degrees. [Method] With the L. chinensis in Inner Mongolia Baiyinxile Ranch as the research object, different rations of nitrogen fertilizer were applied to the grassland （0, 30, 50, 80 g/m^2）. The effect of different gradients of nitrogen fertilizer on photo- synthetic rate of Leymus chinensis, and the effect on grasslands of different degrading degrees were analyzed. [Result] The photosynthetic rate of L. chinensis in- creased with the increase of nitrogen gradients; in the grassland communities with different degrading degrees, the responses of the photosynthetic rate of L. chinensis to nitrogen fertilizer were different, and the response in the grassland with severe degradation was the best. [Conclusion] Nitrogen fertilizer played an important role in enhancing the restoration degree of grassland.

Responses of chlorophyll fluorescence and nitrogen level of Leymus chinensis seedling to changes of soil moisture and temperature

Controlled experiment of Leymus chinensis seedlings grown in the environmental growth chambers at 3 soil moisture levels and 3 temperature levels was conducted in order to improve the understanding how leaf photosynthetic parameters will respond to climatic change. The results indicated that soil drought and high temperature decreased the photochemical efficiency of photosystem(F v/F m), the overall photochemical quantum yield of PSII(yield), the coefficient of photochemical fluorescence quenching(q\-P), but increased the coefficient of non-photochemical fluorescence quenching(q\-N). Severe soil drought would decrease F v/F m and yield by 3.12% and 37.04% under 26℃ condition, respectively, and 6.60% and 73.33% under 32℃ condition, respectively, suggesting that higher temperature may enhance the negative effects of soil drought. All the soil drought treatments resulted in the decline in leaf nitrogen content. There was no significant effect of temperature on leaf nitrogen level, but higher temperature significantly reduced the root nitrogen content and the ratio of root nitrogen to leaf nitrogen, indicating the different strategies of adaptation to soil drought and temperature. It was also implied that higher temperature would enhance the effect of soil drought on leaf photosynthetic capacity, decrease the adaptability of Leymus chinensis to drought.

Effects of Vitamin E on the Activities of Protective Enzymes and Membrane Lipid Peroxidation in Leymus Chinensis under Drought Stress

Leymus chinensis seedlings were treated with 0.05--10 mmol/L vitamin E under osmotic stress in the presence of polyethylene glycol（PEG） as the stress reagent. The effects of the different concentrations of exogenous vitamin E on the activities of SOD, POD and free proline, and the MDA contents under drought stress were examined so as to ascertain the mechanism of Leymus chinensis resistance to drought stress and explore the possible preventive measures. The results indicate that the activities of SOD and POD decreased but the free proline and MDA contents increased as drought stress was accentuated, showing an enhancement of oxidative stress that may cause a decline in membrane stabilization. However, the activities of SOD and POD and the free proline content increased, whereas the MDA content reduced in Leymus chinensis pretreated with vitamin E in comparison with that of the control. This indicates that exogenous vitamin E enhanced the antioxidation of Leymus chinensis seedlings. It suggests that cytomembrane can be protected from damage by increasing the free proline content and the activities of SOD and POD that result in enhancing the drought resistance of Leymus chinensis seedlings.

Dynamics of Zn between soil and plant in Northeast Leymus chinensis grassland

A study was conducted to determine the dynamic of Zn content between soil and plant in the natural protection zone of Leymus chinensis grassland in Changling County （44°30′-44°45′N; 123°31 ′-124°10′E）, Jilin Province, China. Results showed that the total Zn content was lower, available Zn content had a moderate level in the soil, and the plants was not lack of Zn. During the growing season, content of total Zn and available Zn in soil showed a down-trend distribution along the soil profile. Content of total Zn had a significantly positive correlation with that of the organic matter, but it was negatively correlated to soil pH. Monthly dynamic of the average content of total Zn showed a ＂V＂ type curve in the growing season from May to August, and July was the nadir. The trend of the average content of available Zn was similar to the content of total Zn, but was down after August; Zn content variation in the organs and litter of L. chinensis was great, with the order of root〉rhizome 〉leaf〉stem〉litter. The ratio of available Zn content in A layer versus B layer was more than 2 times that of the total Zn, which indicated that the soil of A layer had higher enrichment capacity of available Zn. The enrichment of Zn in the root of L chinensis was 44.17 times as that in the soil. The absorbing intensity of root had a significantly negative correlation with the activity of Zn in the soil （r=-0.8800, p〈0.01）.

Comparative Analysis of Polysaccharides from Two Ecological Types of Leymus chinensis

Leymus chinensis（Trin.） Tzvel.,widely distributed at eastern Eurasian steppe and divided into gray-green type and yellow-green type,has different stress resistance to environment.In the present study,the water-soluble polysaccharides from two ecotypes of L.chinensis were analyzed in detail,and the differences between polysaccharides from the two ecotypes of L.chinensis in the yield,monosaccharide composition,molecular weight and structure were clarified.The polysaccharides of L.chinensis were composed of both neutral and acidic polysaccharides.The neutral polysaccharides contained mannose,glucose,galactose,xylose and arabinose,and mainly consisted of β-1,4-Glcp,α-1,3-Galp and α-1,2-Xylp residues.The acidic polysaccharides contained mannose,rhamnose,glucuronic acid,galacturonic acid,glucose,galactose,xylose and arabinose.However,the yields,monosaccharides contents and the molecular weights of the polysaccharides from the two ecotypes of L.chinensis were different.Moreover,the resistance type（gray-green type） of L.chinensis contained a number of α-1,3-Manp and reducing end of β-Glcp residues,and much more O-methyl groups than normal type（yellow-green type） of L.chinensis.The differences of the polysaccharides of the two ecotypes of L.chinensis might be due to the long-term environmental adaptability of plant,and the differences of the polysaccharides might influence the stress resistance of L.chinensis.

Intra-specific variations of two Leymus chinensis divergence populations in Songnen Plain, Northeast China

Population demography, seed production, biomass allocation, net photosynthesis and transpiration of two Leymus chinensis divergent populations and between two years in Songnen plain, northeast China were compared. Strong differences between the dry 1997 and moist 1998 occurred in vegetative shoot and sexual shoot densities, sexual differentiation and tiller densities, as well as in the lengths of inflorescence, seed numbers per inflorescence, seed weights and biomass allocation in each population respectively ( P < 0.01). While strong differences between the two populations occurred in vegetative shoot densities, sexual shoot densities, sexual differentiation and seed weights in each year ( P < 0.01). The differences between the two populations in tiller densities and in biomass allocation to sexual shoots were significant ( P <0.05). But there were no significant differences between the two populations in the lengths of inflorescence, seed numbers per inflorescence and biomass allocation to rhizomes and vegetative shoots ( P >0.05). Excepting the transpiration rate in the early June, the differences between the two populations in net photosynthesis and transpiration rate of vegetative shoots and sexual shoots were strongly significant in the early June and July respectively ( P <0.01). Relative stable variations in population demography and physiological traits between the two populations indicated that they are divergently in the Songnen Plain.

Effect of Different Proportions of Leymus chinensis in Diet on Plasma Enzymes Activities of Wulong Geese

Eight-month-old Wulong geese were fed with the diet with different proportions of Leymus chiesensis at the same energy and protein level. The activities of plasma Alkaline Phosphatase （AKP）, Acid Phosphatase （ACP）, Choline Esterace （CHE）, Lactic Dehydrogenase （LDH）, Glutamicoxalacetic Transaminase （GOT） and Glutamatepyruvate Transaminase （GPT） were determined and then their correlations with fiber digestibility and the apparent digestibility of Ca, R Met and Cys were analyzed. The results showed that the activity of AKP took on a significant downtrend as the increasing of Leymus chinensis proportion （P〈0.05）. There was a negative correlation between the AKP activity and the digestibility of crude fiber （CF）, neutral detergent fiber （NDF） and acid detergent fiber （ADF） （P〈0.05）. However, there was a significant positive correlation between the AKP activity and the apparent digestibility of Ca, Met and Cys （P〈0.05）. Moreover, the activities of the other five enzymes had no significant correlation with the digestibility of CF, NDF, ADF, and the apparent digestibility of Ca, Met and Cys （P〉0.05）.

Genetic Dissimilarity of Clonal Populations of Leymus chinensis (Trin.) Tzvel in Heterogeneous and Homogeneous Habitats in Southwestern Heilongjiang Province of China

Leymus chinensis （Trin.） Tzvel is the characteristic of the Eastern Asia, and is a perennial herbaceous rhizome species on L. chinensis steppe of the Northern China, the Eastern Mongolia, and area of Outer-Baikal of Russia. It has both vegetative and sexual reproductions, and results in complexity of the population structure. Thus, it is difficult to identify distinct clones. Author used the randomly amplified polymorphic DNA （RAPD） technique to identify the clonal structure of heterogeneous and homogeneous habitats in two populations ofLeymus chinensis, analyzed 260 ramets samples using 12 primers and 60 polymorphie markers （reproducible RAPD bands）. The results proved elonal diversity described using PD, D, and E was higher in the HCS habitat than that in the HPS habitat. The PD, D, and E in the HCS of Songzhan population were 0.21, 0.90, and 0.91, respectively, whereas the PD, D, and E in the HPS of Songzhan population were only 0.09, 0.68, and 0.77, respectively; the PD, D, and E in the HCS of Yongle population were 0.19, 0.92 mad 0.90, respectively, correspondingly, these measures in the HPS of Yongle population were only 0.09, 0.63, and 0.79, respectively. The results also indicated that clonal population ofL. chinensis in HPS habitat had lower genetic dissimilarity index than that in the HCS habitat.

Seed Germination and Callus Induction in Leymus chinensis of Different Seed Colors

[ Objective] To explore the effects of different seed colors on seed germination and callus induction of Leymus chinensis. [ Method ] According to the color of mature seeds of Leymus chinensis, they were divided three color types, namely, pale yellow, yellow and black. Their seed germination and callus induction were observed. [ Result] The pale yellow Leymus chinensis seeds had higher germination rate （32.5%） and rate of callus induction （28.5%）. Based on the results of the TI＇C method and electrical conductivity, the seed viability was decreased with the deepening seed color. [ Conclusion] The study laid the foundation for establishing highly efficient plant regeneration system of Leymus chinensis.

Influence of litter layer on ecological environment in northeastern leymus chinensis grassland of China

In this paper, we adopted simulation method to discuss influences of litter layer on plants habitat in grassland. Results indicated that ground surface evaporation, soil moisture, surface temperature, soil pH, soil bulk density and soil porosity were all strongly related to the litter quantity. Potassium （K） and organic materials in the soil covered by litter layer were higher than those in the soil uncovered by litter layer. With 100 g.ln-z increase of litter, the percentage of organic materials increased by 17.9%, nitrogen （N） increased by 7.6%, phosphor （P） increased by 26.4%, and K increased by 3.8%. With the litter accumulation amounting up to 600 g-m-2, the percentage of organic materials increased by 1.8 times, N increased by 81.5%, P increased by 1.8 times and K increased by 26.4%. According to the expected coefficient method of optimization, a mathematical model was established about the optimal accumulation quantity of litter.