There are two 5 '-flanking regions of bkt encoding beta-carotene ketolase in Haematococcus pluvialis

The unicellular green alga Haematococcus pluvialis accumulates a commercially valuable astaxanthin, with levels reaching up to 4% dry weight under environmental stress. In recent years, much effort has been devoted to understanding the molecular mechanisms regulating astaxanthin biosynthetic pathways. Beta-carotene ketolase (bkt), with control being exhibited at the transcription level, plays an important role in astaxanthin biosynthesis by H. pluvialis. Here we demonstrate the presence of two separate 5'-flanking regions [1.5 kilobase (kb) and 2 kb] of bkt (bkt1 and bkt2) that possess regulatory elements similar to those of known stress-responsive genes in plants. Results of 5'-deletion constructs and transient beta-galactosidase expression assays demonstrate that there may be positive regulatory elements governing expression in the shorter promoter at -1060/-900 from the 1.5 kb 5' region, and in the longer promoter at -1838/-1219 and at -1046/ -734 from the 2 kb 5' region relative to each homologous ATG start codon. Furthermore, our present studies reveal that the first intron (+371/+497) downstream from the 1.5 kb 5' untranslated region of bkt1 may function as a negative regulatory element to regulate its own promoter.

Deep Fractionation of Clinopyroxene in the East Pacific Rise 13 degrees N: Evidence from High MgO MORB and Melt Inclusions

Mid-ocean ridge basalts (MORBs) from East Pacific Rise (EPR) 13 degrees N are analysed for major and trace elements, both of which show a continuous evolving trend. Positive MgO-Al2O3 and negative MgO-Sc relationships manifest the cotectic crystallization of plagioclase and olivine, which exist with the presence of plagioclase and olivine phenocrysts and the absence of clinopyroxene phenocrysts. However, the fractionation of clinopyroxene is proven by the positive correlation of MgO and CaO. Thus, MORB samples are believed to show a "clinopyroxene paradox". The highest magnesium-bearing MORB sample E13-3B (MgO = 9.52%) is modelled for isobaric crystallization with COMAGMAT at different pressures. Observed CaO/Al2O3 ratios can be derived from E13-3B only by fractional crystallization at pressure > 4 +/- 1 kbar, which necessitates clinopyroxene crystallization and is not consistent with cotectic crystallization of olivine plus plagioclase in the magma chamber (at pressure similar to 1 kbar). The initial compositions of the melt inclusions, which could represent potential parental magmas, are reconstructed by correcting for post-entrapment crystallization (PEC). The simulated crystallization of initial melt inclusions also produce observed CaO/Al2O3 ratios only at > 4 +/- 1 kbar, in which clinopyroxene takes part in crystallization. It is suggested that MORB magmas have experienced clinopyroxene fractionation in the lower crust, in and below the Moho transition zone. The MORB magmas have experienced transition from clinopyroxene+plagioclase+olivine crystallization at > 4 +/- 1 kbar to mainly olivine+plagioclase crystallization at < 1 kbar, which contributes to the explanation of the "clinopyroxene paradox".

Generation and evolution of magma beneath the East Pacific Rise: Constraints from U-series disequilibrium and plagioclase-hosted melt inclusions

Mid-ocean ridge basalt (MORB) samples from the East Pacific Rise (EPR 12 degrees 50'N) were analyzed for U-series isotopes and compositions of plagioclase-hosted melt inclusions. The Ra-226 and Th-230 excesses are negatively correlated; the Ra-226 excess is positively correlated with Mg# and Sm/Nd, and is negatively correlated with La/Sm and Fe-8; the Th-230 excess is positively correlated with Fe-8 and La/Sm and is negatively correlated with Mg# and Sm/Nd. Interpretation of these correlations is critical for understanding the magmatic process. There are two models (the dynamic model and the "two-porosity" model) for interpreting these correlations, however, some crucial parameters used in these models are not ascertained. We propose instead a model to explain the U-series isotopic compositions based on the control of melt density variation. For melting either peridotite or the "marble-cake" mantle, the FeOt content, Th-230 excess and La/Sm ratio increases and Sm/Nd decreases with increasing pressure. A deep melt will evolve to a higher density and lower Mg# than a shallow melt, the former corresponds to a long residence time, which lowers the Ra-226 excess significantly. This model is supported by the existence of low Ra-226 excesses and high Th-230 excesses in MORBs having a high Fe-8 content and high density. The positive correlation of Ra-226 excess and magma liquidus temperature implies that the shallow melt is cooled less than the deep melt due to its low density and short residence time. The correlations among Fe-8, Ti-8 and Ca-8/Al-8 in plagioclase-hosted melt inclusions further prove that MORBs are formed from melts having a negative correlation in melting depths and degrees. The negative correlation of Ra-226 excess vs. chemical diversity index (standard deviation of Fe-8, Ti-8 and Ca-8/Al-8) of the melt inclusions is in accordance with the influence of a density-controlled magma residence time. We conclude that the magma density variation exerts significant control on residence time and U-series isotopic compositions. (c) 2010 Elsevier B.V. All rights reserved.

中国对虾育种方法研究与遗传连锁图谱的构建

本研究以中国对虾为材料，以杂交育种和选择育种为目标，进行了系统的中国对虾杂交育种试验、生长性状遗传参数试验及其分子遗传连锁图谱的构建工作。结果表明以不同地理群体杂交作为基础群体，然后采用系统的选择育种方法可以获得较好的选择效果。构建的遗传连锁图谱为中国对虾分子辅助育种提供一定的基础。这些试验结果为中国对虾合理系统的育种工作提供了理论基础和数据支持。其具体结果如下： 1. 试验对中国对虾黄渤海水域乳山湾群体（WYP）和朝鲜半岛南海群体（WKN）的2个群体及其杂交后代不同月龄生长情况和存活率进行了研究，测量体长（TL）、头胸甲长（CL）、头胸甲宽（CW）、第2、3腹节高（HST）、第2、3腹节宽（WST）、体重（BW）和存活率共7个性状，计算各项指标的杂种优势率，并对各性状进行了方差分析和多重比较。其3月龄生长情况和存活率研究结果表明，存活率在乳山湾群体（WYP♀）× 朝鲜半岛南海群体（WKN♂）杂交后代出现杂种劣势外，其他指标都表现出不同程度的杂种优势(4.37%～23.96% )。除了存活率外，杂交后代生长性状均显著高于亲本，乳山湾群体（WYP♀）×朝鲜半岛南海群体（WKN♂）杂交后代高于朝鲜半岛南海群体（WKN♀）× 乳山湾群体（WYP♂）杂交后代，黄渤海水域乳山湾群体高于朝鲜半岛南海群体后代。为确定测量性状与中国对虾体重的相关程度，建立了用体长(X1)，头胸甲宽(X2)，第2、3腹节宽(X3)，头胸甲长(X4)，第2、3腹节高(X5)估计体重的多元回归方程：Y = －2.056 + 0.03X1 + 0.076X2 + 0.078X3 + 0.033X4 + 0.043X5。 2. 中国对虾黄渤海水域乳山湾群体（WYP）和朝鲜半岛南海群体（WKN）2个群体及其杂交后代在4月龄时期的6个生长指标和存活率的杂种优势范围在0.514%到14.95%之间，WYP♀×WKN♂杂交后代在这7个指标中都高于WKN♀×WYP♂杂交后代。5月龄杂交后代也表现出一定程度的杂种优势，其范围在-9.000%～19.090%之间，但头胸甲长、第2、3腹节处高和存活率3个指标出现杂种劣势。不同杂交组合各个阶段生长发育情况和存活率在杂种优势表现出一定的规律。随着月龄的增加，WKN♀×WYP♂杂交后代杂种优势率有所增加，而WYP♀×WKN♂杂交后代的却有所降低。ANOVA分析结果表明，杂交后代在存活率方面与双亲差异不显著。4月龄的分析结果发现杂交后代在WST和BW这2个指标上存在显著差异。LSD多重比较结果显示，WYP♀×WKN♂杂交后代在BW指标上与亲本存在显著差异，在WST指标方面与其他3个组合的后代差异显著。5月龄的数据分析结果发现，杂交后代除体重存在显著差异外，其他各项指标差异均不显著。LSD多重比较结果发现，WKN♀×WYP♂杂交后代体重与其亲本WKN存在显著差异。 3. 对2个野生群体——朝鲜半岛南海岸群体（WKN）和黄渤海群体（WYP）和3个养殖群体——朝鲜半岛群体的养殖一代（FKN），黄海1号（HH1）和即抗98（JK98）进行杂交试验的研究，结果表明JK98 (♀)  WKN (♂)组合在存活率方面最高，其余的依次为WYP (♀) WKN (♂)，WKN (♀) WYP (♂)，FKN (♀)HH1 (♂) 和 WYP (♀) FKN (♂)。而在体重方面FKN(♀)  HH1(♂)组合最高，其余的依次为WKN (♀) WYP (♂)，WYP (♀) WKN (♂)，WYP (♀)FKN (♂) 和 JK98 (♀)WKN (♂)。在所有生长性状方面，JK98 (♀)  WKN (♂)在5个组合中是最低的。方差检测结果表明，TL、CL、HST、LL和BW这5个指标在不同组合间存在差异，而其他指标不存在差异。多重比较结果发现JK98 (♀)WKN (♂)组合的TL与其他组合间差异极显著，HST指标与WKN (♀) WYP (♂)，FKN(♀)  HH1(♂)和 WYP (♀)  WKN (♂)这3个组合差异显著，BW指标与WKN (♀) WYP (♂) 和 FKN(♀)  HH1(♂)差异显著。 4. 通过人工授精的方法建立了中国对虾21个半同胞家系，测量了中国对虾21个半同胞（46个全同胞）家系的TL、CL、CW、HST、WST、第1腹节长（FL）、第6腹节长（LL）。利用MTDFREML软件得到生长性状遗传力在0.15~0.35之间，属于中度遗传力范围。TL的遗传力为0.34±0.071，CL的为0.30±0.070，CW为0.35±0.077，WST为0.33±0.073，HST为0.33±0.073，FL的最低为0.15±0.044，LL的为0.24±0.059。各个性状间表现出高的正相关，其中CW和TL以及HST的遗传相关最大，FL和WST的遗传相关最小。 通过以上杂交育种和选择育种的研究，认为单纯的依靠杂交育种来改善中国对虾的育种工作可能具有一定的局限性。所以在实际的育种过程中，以中国对虾不同群体的杂交后代作为基础群体，并以此为基础进行系统的选择育种应该具有更大的潜力。 5. 本试验利用中国对虾F2群体和AFLP分子标记技术进行了遗传连锁图谱的构建。利用55对AFLP引物组合对F2家系的110个个体进行了研究，结果检测到532个符合作图策略的AFLP标记。利用卡方检验检测分离位点是否符合孟德尔分离定律。对于符合3：1比例的分离位点利用F2自交模型构建性别平均连锁图谱，对于符合1：1比例的分离位点利用拟测交理论分别构建中国对虾的雌性和雄性遗传连锁图谱。雌性、雄性和性别平均遗传图谱分别有28、35和44个连锁群，图谱实际长度分别为1090、1617和1772.1 cM。中国对虾遗传连锁图谱估计基因组长度为2420 cM，符合与人类基因组相比的对虾类基因组长度。中国对虾雄性遗传连锁图谱比雌性遗传连锁图谱长32.6%，这可能说明中国对虾不同性别存在不同的重组率。通过皮尔逊相关系数检测认为AFLP标记在中国对虾图谱上分布均匀。本文利用AFLP标记构建的中国对虾遗传连锁图谱为中国对虾基因组研究和遗传改良提供一定的基础，同时也应该开发微卫星等共显性标记，为遗传连锁图谱的整合提供条件。

东太平洋海隆13°N和大西洋Logatchev热液区附近热液柱的研究

海底热液柱的研究为我们认识海底热液活动提供了一个放大镜，为测定控制各种热液种类最终散布的化学反应提供了天然实验室，是我们认识海底热液活动环境效应、寻找海底热液多金属硫化物资源、定位新的热液喷口区、表征和定量海底热液活动释放的有利工具。本文在海底对热液柱研究概况了解的基础上总结了海底热液柱的类型、运动特征、影响其扩展的因素及其地球化学特征；在对2003年11月DY105-12、14航次第六航段和2005年10月DY105-17A航次大西洋航段，在EPR12°39′N～12°54′N区域和MAR Logatchev热液区（MAR 14°45＇N）获得的CTD数据分析的基础上，讨论了在东太平洋海隆和大西洋Logatchev热液区出现的温度异常；在对EPR12°39′N～12°54′N区域内所采水样地球化学数据分析的基础上，探讨了EPR12°39′N～12°54′N上方海底热液柱的地球化学特征及其影响因素，发现该调查区内Mg亏损可能由热液流体和海水混合达到平衡后上浮的一种贫Mg液体引起，Ca亏损是一种或几种因素共同作用的结果，其中最主要的原因可能是富二氧化碳流体夹带周围海水上升过程中形成碳酸钙颗粒，从而引起Ca的减小，Cl、Br富集可能是热液活动引起相分离后后期卤水注入海水的结果。

东太平洋海隆13°N附近沉积物元素地球化学研究

对东太平洋海隆（EPR）13°N西侧2个沉积物岩芯进行了碳酸盐、常量和微量元素含量测定。E271站位沉积物中Fe含量在8.5~13.8%之间，Mn含量在1.7~3.17%之间；E272站位Fe 5.7~13%，Mn 0.12~3.31%，沉积物中高铁锰含量显示在热液柱颗粒物质在EPR13°N西翼沉降明显。E271站位CaCO3含量在5.9~27.6%之间，E272站位CaCO3含量在6.7~38.2%之间，在两个站位沉积物柱样中发现少量有孔虫化石。Cu、Pb和Zn因为热液柱颗粒物质沉降，在沉积物中富集。Cu/Fe，Pb/Fe和Zn/Fe比值比热液喷口处颗粒物中低，Pb表现出在海水中运移距离较短或者是只有少量Pb随热液柱扩散运移。热液柱沉降作用使Li，Mo和Ni在沉积物中富集。此外，铁锰氧化物颗粒在海水中吸附-共沉淀作用导致V在沉积物中富集。沉积物中Ti和Al有非常好的相关性，其Ti/Al比值为0.05左右。U在岩芯中含量与深海沉积相似。

Feeding and egg production of the planktonic copepod Calanus sinicus in spring and autumn in the Yellow Sea, China

Shipboard incubations were conducted in spring (April) and autumn (October/November) 2006 to measure the feeding and egg production rates (EPR) of Calanus sinicus in the Yellow Sea, China. The ingestion rate (2.08-11.46 and 0.26-3.70 mu g C female(-1) day(-1) in spring and autumn, respectively) was positively correlated with microplankton carbon concentrations. In the northern part of the Yellow Sea, feeding on microplankton easily covers the respiratory and production requirements, whereas in the southern part in spring and in the frontal zone in autumn, C. sinicus must ingest alternative food sources. Low ingestion rates, no egg production and the dominance of the fifth copepodite (CV) stage indicated that C. sinicus was in quiescence inside the Yellow Sea Cold Bottom Water (YSCBW) area in autumn. Calanus sinicus ingested ciliates preferentially over other components of the microplankton. The EPR (0.16-12.6 eggs female(-1) day(-1) in spring and 11.4 eggs female(-1) day(-1) at only one station in autumn) increased with ciliate standing stock. Gross growth efficiency (GGE) was 13.4% (3-39%) in spring, which was correlated with the proportion of ciliates in the diet. These results indicate that ciliates have higher nutrient quality than other food items, but the low GGE indicates that the diet of C. sinicus is nutritionally incomplete.

Seasonal variation of reproduction rates and body size of Calanus sinicus in the Southern Yellow Sea, China

Annual variations of egg production rate (EPR) and clutch size of Calanus sinicus, as well as body size of females (prosome length and dry weight), were investigated at a series of stations in the Southern Yellow Sea by onboard incubation. Calanus sinicus was spawning in all the 11 cruises investigated, and the annual variation of EPR was bimodal. Monthly average EPR was highest from May to July, respectively, 5.97, 5.36 and 6.30 eggs female(-1) d(-1), then decreased dramatically to only 1.37 eggs female(-1) d(-1) in August and attained the lowest 1.07 eggs female(-1) d(-1) in October. In November, average EPR increased again to 4.31 eggs female(-1) d(-1). Seasonal variation of clutch size was similar to EPR, except that it decreased gradually after August rather than dramatically as did EPR. Prosome length of females was maximum in May and minimum in October, but dry weight was highest in November. Monthly average EPR correlated better with prosome length than dry weight, while clutch size was rather determined by dry weight of females. It is suggested that egg production of C. sinicus was active during two discontinuous periods when both surface and bottom temperature fell into its favorite range (i.e. 10-23degreesC), and different reproductive strategies were adopted in these two reproductive peaks: other than the highest EPR, longer prosome length was also achieved by C. sinicus from May to July, while females in November developed shorter bodies but accumulated more energy for reproduction.

Spring and autumn reproduction of Calanus sinicus in the Yellow Sea

The fecundity, hatching success and naupliar survival of Calanus sinicus were studied in the Yellow Sea during research cruises in April and October 2006, with emphasis on the regulation of reproduction. During both cruises, the egg production rate (EPR) showed large spatial variations (0 to 25.4 eggs female(-1) d(-1)), generally coinciding with the food availability. In April, the abundant phytoplankton and ciliates in the study area supported active reproduction, which would probably initiate the annual population development. In October, females remained immature in the Yellow Sea Cold Water Mass, likely due to the unfavorable environment (poor food and low temperature). However, reproduction and recruitment were high in the neritic region, which may explain the local population recovery in late autumn. Hatching success varied markedly among stations in April (4 to 85%), whereas it was high overall in the neritic region during October (> 90%). Based on the potential recruitment rates, the spring recruitment would be more important for the annual population dynamics, Female gonad maturity, body size and lipid reserve were examined in relation to fecundity. Regression analyses suggested that the reproductive index (defined as the proportion of females with mature gonads) could be an indicator for the EPR of C. sinicus. Among the regulating factors, external food (ciliates and phytoplankton) seems essential for reproduction, whereas inner lipid reserve may mainly serve metabolic needs. Moreover, fecundity is positively related to body size but independent of temperature, which might exert indirect influences on reproduction.

Evolution of nutrient structure and phytoplankton composition in the Jiaozhou Bay ecosystem

The inventories of nutrients in the surface water and large phytoplankton( > 69 pm) were analyzed from the data set of JERS ecological database about a typical coastal waters, the Jiaozhou Bay, China, from 1960s for N, P and from 1980s; for Si. By examining long-term changes of nutrient concentration, calculating stoichiometric balance, and comparing diatom composition, Si limitation of diatom production was found to be more possible. The possibility of Si limitation was from 37% in 1980s to 50% in 1990s. Jiaozhou Bay ecosystem is becoming serious eutrophication, with notable increase of NO2-N, NO3-N and NH4-N from 0.1417 mumol/L, 0.5414 mumol/L, 1.7222 mumol/L in 1960s to 0.9551 mumol/L, 3.001 mumol/L, 8.0359 mumol/L in late 1990s respectively and prominent decrease of Si from 4.2614 mumol/L in 1980s to 1.5861 mumol/L in late 1990s; the nutrient structure is controlled by nitrogen; the main limiting nutrient is probably silicon; because of the Si limitation the phytoplankton community structure has changed drastically.

Seasonal and spatial variation in abundance and egg production of Paracalanus parvus (Copepoda: Calanoida) in/out Jiaozhou Bay, China

The spatial distribution of stage-specific abundance and reproduction of the copepod Paracalanus parvus were studied from October 2005 to September 2006 in the Jiaozhou Bay. This copepod occurred continuously in this bay throughout the year. The species reached the lowest abundance in April and peaked in June. From October to December, distribution center mainly occurred in offshore water and at the mouth of the bay. In winter, early copepodites and adults gradually decreased and till February, most of the population was only comprised of CIV-CV stages. Overwintering copepodites matured in March and males tended to mature before female. From May to September, each stage occurred in the population and gradually reached high abundance. Temperature and chlorophyll a (Chl-a) concentration in the three stations can't clearly explain the seasonal variation in stage-specific abundance, so we surmised the important effect of the Yellow Sea. Egg production rate (EPR) reached its lowest in winter and peaked in June at 60.8 eggs female(-1) day(-1) in nearshore water. In the warming period, EPR in nearshore water was statistically higher and EPR > 10 eggs female(-1) day(-1) lasted longer than that in offshore water, showing the importance of nearshore water for recruitment of R parvus. Our study showed that EPR was positively related to temperature and total chlorophyll a in offshore water and mouth of the bay. In nearshore water, the relationships between EPR and temperature and Chl-a in three size fractions were not the same as those in offshore water, suggesting complicated ecosystem in such a eutrophic area in warming period. (C) 2008 Elsevier Ltd. All rights reserved.

Feeding habits of Calanus sinicus (Crustacea : Copepoda) during spring and autumn in the Bohai Sea studied with the herbivore index

Pigment ingestion rate (PIR) and egg production rate (EPR) of the dominant copepod Calanus sinicus, as well as chlorophyll-a concentration and phytoplankton assemblages were measured in the Bohai Sea, North China in June 1997, October 1998 and May 1999. A herbivore index (H) was also calculated as the carbon specific ratio of PIR and EPR, in order to investigate its feeding habits in the spring and autumn phytoplankton bloom respectively. On average, chlorophyll-a concentration was relatively similar (1-1.34 mg m(-3)) in the three cruises, but PIR was quite different. It was 3.24 mu g C female(-1) d(-1) in October, equivalent to one half of the PIR for June and one third of the PIR for May. Average EPR was highest in May, and quite similar during the other two months. According to H values, herbivorous feeding contributed 100% of the egg production of C. sinicus in June, 82.5% in May, but only 47.8% in October. It is possible that omnivorous feeding of C. sinicus in October was induced by a prevalence of large-sized diatoms and sufficient non-phytoplankton food resources during the autumn bloom period.

Genetic improvement on Chinese shrimp (Fenneropenaeus chinensis): growth and viability performance in F1 hybrids of different populations

Fenneropenaeus chinensis distributed in the Yellow Sea and Bohai Sea of China and the west coast of the Korean Peninsula. Different geographical populations represent potentially different genetic resources. To learn further the characteristics of different geographical population, crosses among two wild and three farmed populations were produced. The two wild populations were from the Yellow Sea and Bohai Sea (WYP), and the west coast of the Korean Peninsula and coast (WKN). The three farmed populations included the offspring of first generation of wild shrimp from coast in Korea (FKN), the Huang Hai (the Yellow Sea in Chinese) No.1 (HH1), and JK98. The phenotypes growth and survival rates of these populations were compared to confirm the feasibility for crossbreeding. The body length (BL), carapace length (CL), carapace width (CW), height of the second and third abdominal segment (HST), width of the second and third abdominal segment (WST), length of the first abdominal segment (LF), length of the last abdominal segment (LL), live body weight (BW), and survival rate were measured. Different combinations were statistically performed with ANOVA and Duncan's Multiple Range Test. The results show that the survival rate of JK98(a (TM) Euro)xWKN(a (TM),) was the highest, followed by WYP(a (TM) Euro)xWKN(a (TM),), FKN(a (TM) Euro)xWYP(a (TM),), FKN(a (TM) Euro)xHH1(a (TM),) and WYP(a (TM) Euro)xFKN(a (TM),); the body weight of FKN(a (TM) Euro)sxHH1(a (TM),) was the highest, followed by FKN(a (TM) Euro)xWYP(a (TM),), WYP(a (TM) Euro)xWKN(a (TM),), WYP(a (TM) Euro)xFKN(a (TM),) and JK98(a (TM) Euro)xWKN(a (TM),); the total length had the same ranking as the body weight. All growth traits in hybrids JK98(a (TM) Euro)xWKN(a (TM),) were the lowest among all combinations. F1 hybrids had significant difference (P < 0.05) in BL, CL, HST, LL, and BW; and insignificant difference (P > 0.05) in other growth traits and survival rate. The results of Duncan's Multiple Range Test are that BL and CL of JK98(a (TM) Euro)xWKN(a (TM),) were significantly different from the other combinations; HST different from the combination of FKN(a (TM) Euro)xWYP(a (TM),), FKN(a (TM) Euro)xHH1(a (TM),) and WYP(a (TM) Euro)xWKN(a (TM),); and BW different from FKN(a (TM) Euro)xWYP(a (TM),) and FKN(a (TM) Euro)xHH1(a (TM),). As a whole, the results indicate that the FKN(a (TM) Euro)xHH1(a (TM),) was the best combination in all growth traits. Therefore, hybridization can introduce the variation to base populations. The systematic selection program based on additive genetic performance may be more effective than crossbreeding.

Effect of N fertilizer on the productivity and nutritive values of perennial grass mixtures in the alpine region of Qinghai-Tibetan Plateau, China

A study was conducted on grass mixtures that included smooth bromegrass (SB) + drooping wild ryegrass (DW), smooth bromegrass + Siberian wild ryegrass (SW) + crested wheatgrass (CW) and smooth bromegrass + Siberian wild ryegrass + drooping wild ryegrass + crested wheatgrass in the alpine region of Qinghai-Tibetan Plateau. The study was conducted from 1998 to 2000 to investigate the effects of N application rates and growing year on herbage dry matter (DM) yield and nutritive values. Herbage DM production increased linearly with N application rates. The effect of N application on DM yields was greater (P < 0.05) in the 2nd and 3rd production years than in the establishment year. Dry matter yields of SB + SW + CW and SB + SW + DW + CW can reach as high as 15 000 kg ha(-1) at 345 kg ha(-1) N rate in the 3rd growing year. With increased N application rates, crude protein (CP) contents and 48 h in sacco DM degradability of grasses increased (P < 0.05). No effect (P > 0.05) of N application was detected on organic matter (OM) and acid detergent fibre (ADF) concentration. It can be concluded that for increased biomass production in the alpine region of the Qinghai-Tibetan Plateau, a minimum of 345 kg N ha(-1) should be applied to grass stands in three split application of 115 kg N ha(-1), in early June, early July and late July