青稞胆碱单加氧酶基因cDNA全长开放阅读框克隆及序列分析

甜菜碱是一种存在于多种生物体内的季胺类高效渗透调节剂,是由甜菜碱合成酶系催化的。为了研究甜菜碱合成酶系基因的表达和青稞高抗逆性之间的关系,我们以经200mmol/L NaCl预处理72h的青稞幼苗叶片及根中提取的总RNA为模板,采用RT-PCR技术,扩增得到全长1224bp,推测编码407个氨基酸残基的青稞胆碱单加氧酶(choline monooxygenase,cmo)基因cDNA全长开放阅读框(open reading frame,ORF),将其命名为hvcmo1并在GenBank中注册,获得登录号为GU810840。生物信息学分析表明,推定的氨基酸序列中含有与已报道高等植物胆碱单加氧酶氨基酸序列中的Rieske型铁硫簇结合区和单核铁结合基序等特征性结构域具有高度保守性的特征性结构域。与多种高等植物cmo基因进行序列同源性比对分析结果显示:hvcmo1的核苷酸序列与甜菜、盐角草、菠菜、辽宁碱蓬、拟南芥、玉米、水稻、高粱、羊草和大麦等植物的cmo mRNA编码区相似性分别为55.3%、55.4%、55.6%、5.9%、61.3%、82.6%、83.3%、83.4%、95.5%和99.5%。实验过程中,我们发现,青稞hvcmo1基因的表达必须被一定浓度的NaCl所诱导,说明hvcmo1可能在调节环境胁迫下青稞细胞内甘氨酸甜菜碱合成方面起着关键的作用,同时它可以作为研究环境胁迫下甘氨酸甜菜碱合成调节的模型。本结果为研究青稞甘氨酸甜菜碱合成酶系基因的诱导表达及其与青稞强抗逆性之间的关系奠定了基础。     

72份青稞氨基酸组成与营养价值评价

为了解青稞氨基酸的组成及营养品质,本研究测定了72份青稞材料籽粒的17种氨基酸(色氨酸除外)含量,通过对供试青稞材料氨基酸含量进行聚类分析,并比较育成品种与地方品种的氨基酸组成差异,对青稞的氨基酸进行了营养价值评价。结果表明,青稞氨基酸总含量平均为87.454 mg/g DW,变幅为47.8~178.7 mg/g DW,其中必需氨基酸含量占37.15%,谷氨酸的含量最高且变异大,蛋氨酸含量最低,赖氨酸含量变异小,91.67%的青稞材料的第一限制性氨基酸为赖氨酸。青稞氨基酸的贴近度为0.903,与世界粮农组织(FAO)、世界卫生组织(WHO)、联合国大学(UNU)提出的模式蛋白质的必需氨基酸组成较接近,氨基酸比值系数分SRC(73.14)较高。非必需氨基酸占青稞总氨基酸含量的62.85%,其中鲜味和甜味氨基酸含量分别为26.58 mg/g DW和21.85 mg/g DW,分别占总氨基酸含量的30.04%和24.43%。不同青稞材料的氨基酸含量和营养价值有很大的差异,地方品种各氨基酸含量均高于育成品种。供试材料中有4份青稞的氨基酸营养价值及风味氨基酸含量均较高。研究结果可为优质氨基酸组成的青稞选育及青稞加工提供指导。     

New material ofChalicotherium from the Tsaidam Basin in the northern Qinghai-Tibetan Plateau, China

New material of a chalicothere was recently found from Tsaidam Basin, the only late Tertiary locality within the northern Qinghai-Tibetan Plateau established byBirger Bohlin during the Sino-Swedish Expeditions in 1927–33. A partial lower jaw with two molars helps to identify it asChalicotherium cf.C. brevirostris (Colbert), a species found in Chinese Tunggurian age (middle Miocene). Occurrence ofChalicotherium in the Tsaidam Basin indicates the presence of a wooded environment along freshwater streams, a condition much milder than the desert environment at the present time. The Tsaidam fauna thus provides a reference point in the current debate on the timing of environmental deterioration during the uplift of Tibetan Plateau.

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Development of a Core Set from a Large Rice Collection using a Modified Heuristic Algorithm to Retain Maximum Diversity

A new heuristic approach was undertaken for the establishment of a core set for the diversity research of rice. As a result, 107 entries were selected from the 10 368 characterized accessions. The core set derived using this new approach provided a good representation of the characterized accessions present in the entire collection. No significant differences for the mean, range, standard deviation and coefficient of variation of each trait were observed between the core and existing collections. We also compared the diversity of core sets established using this Heuristic Core Collection （HCC） approach with those of core sets established using the conventional clustering methods. This modified heuristic algorithm can also be used to select genotype data with allelic richness and reduced redundancy, and to facilitate management and use of large collections of plant genetic resources in a more efficient way.     

Carbon Dioxide Dynamics and Controls in a Deep-water Wetland on the Qinghai-Tibetan Plateau

To initially characterize the dynamics and environmental controls of CO₂, ecosystem CO₂ fluxes were measured for different vegetation zones in a deep-water wetland on the Qinghai-Tibetan Plateau during the growing season of 2002. Four zones of vegetation along a gradient from shallow to deep water were dominated, respectively by the emergent species Carex allivescens V. Krez., Scirpus distigmaticus L., Hippuris vulgaris L., and the submerged species Potamogeton pectinatus L. Gross primary production (GPP), ecosystem respiration (Re), and net ecosystem production (NEP) were markedly different among the vegetation zones, with lower Re and GPP in deeper water. NEP was highest in the Scirpus-dominated zone with moderate water depth, but lowest in the Potamogeton-zone that occupied approximately 75% of the total wetland area. Diurnal variation in CO₂ flux was highly correlated with variation in light intensity and soil temperature. The relationship between CO₂ flux and these environmental variables varied among the vegetation zones. Seasonal CO₂ fluxes, including GPP, Re, and NEP, were strongly correlated with aboveground biomass, which was in turn determined by water depth. In the early growing season, temperature sensitivity (Q₁₀) for Re varied from 6.0 to 8.9 depending on vegetation zone. Q₁₀ decreased in the late growing season. Estimated NEP for the whole deep-water wetland over the growing season was 24 g C m⁻². Our results suggest that water depth is the major environmental control of seasonal variation in CO₂ flux, whereas photosynthetic photon flux density (PPFD) controls diurnal dynamics.     

超临界CO_2萃取青稞麸皮油对高血脂症大鼠降脂作用研究

为进一步开发利用青稞麸皮油,研究超临界CO2(SC-CO2)萃取青稞麸皮油对高血脂症大鼠的降脂作用。SC-CO2萃取青稞麸皮油得率为18.53%;用GC-MS分析其脂肪酸成分,主要脂肪酸为亚油酸(59.57%)、十八碳烯酸(17.57%)、棕榈酸(15.07%)。通过饲喂高脂饲料建立高血脂症模型大鼠并同时用青稞麸皮油灌胃,三周后测定大鼠血清TG、TC、LDL-C、HDL-C和AI。SC-CO2萃取青稞麸皮油显著(P<0.05或P<0.01)降低高血脂症大鼠TG、TC、AI水平。说明SC-CO2萃取青稞麸皮油对实验性高血脂症大鼠的血脂有很明显的调节作用,可预防动脉粥样硬化的发生,为实现青稞麸皮油的产业化研究奠定实验基础。     

青藏高原高山流石滩特有植物绵参的谱系地理学研究

绵参(Eriophyton wallichii)是中国青藏高原地区高山流石滩特有植物.为探讨第四纪冰期气候波动对青藏高原高山流石滩植物居群遗传结构以及谱系历史的影响,检测了绵参20个居群187个个体的核糖体内转录间隔区(ITS)的序列变异,共发现19个单倍型,且结果表明大部分居群拥有独自的单倍型.AMOVA分析表明,居群间的变异占了总变异量的89.54％,居群遗传分化指数很高(GST=0.863,NST=0.957),但无明显的谱系结构.我们推测这种独特的谱系地理结构可能主要是由于高山流石滩独特的地形和环境及“孤岛效应”形成的,与先前研究过的青藏高原植物不同,绵参呈现出高度分化的遗传结构,常常呈现出“一个居群,一种单倍型”的模式,这可能是由于第四纪冰期及间冰期尽管绵参的分布范围有扩大和收缩,但始终由于特殊的高山生境和流石滩基质,导致了该物种长期以来居群间很少有基因交流并一直处于隔离状态,造成了现在这样的谱系地理结构.     

Distribution of carbon isotope composition of modern soils on the Qinghai-Tibetan Plateau

This paper presents a large data set on carbon isotope composition (¹³C) of modern soils which were collected under the main vegetation communities along an altitude of 1250–5500m above sea level in the Qinghai-Tibetan Plateau. The ¹³C values of 198 samples range from –28.6 to –15.1 versus PDB and exhibit a clean relation to different vegetation communities from forest (–25.9±1.2) to shrub (–24.7±1.4), steppe (–23.1±1.3), alpine meadow (–23.6±0.7), alpine desert steppe (–21.3±1.6), and alpine desert (–18.9±2.5). We attributed the observed variability in ¹³C values to that the mean annual precipitation (MAP) and the mean annual temperature (MAT) are the main factors controlling the distribution of vegetation types in the Tibetan Plateau, which causes the change in carbon isotope composition of modern soils at any given altitude. The result of both linear and nonlinear regression analyses also confirms that MAP and MAT are the major factors affecting the ¹³C values of surface soils. In the absence of favorable moisture and temperature conditions, low pCO₂ alone is not sufficient to cause the distinct changes in carbon isotope composition of modern soils in the Tibetan Plateau. This study provides some fundamental information on the carbon isotope composition of terrestrial carbon pools and bears some practical significance for the use of carbon isotope data to document vegetation changes and environmental conditions of the high plateau in the past.     

Indehiscent sporangia enable the accumulation of local fern diversity at the Qinghai-Tibetan Plateau

ABSTRACT: BACKGROUND: Indehiscent sporangia are reported for only a few of derived leptosporangiate ferns. Their evolution has been likely caused by conditions in which promotion of self-fertilization is an evolutionary advantageous strategy such as the colonization of isolated regions and responds to stressful habitat conditions. The Lepisorus clathratus complex provides the opportunity to test this hypothesis because these derived ferns include specimens with regular dehiscent and irregular indehiscent sporangia. The latter occurs preferably in well-defined regions in the Himalaya. Previous studies have shown evidence for multiple origins of indehiscent sporangia and the persistence of populations with indehiscent sporangia at extreme altitudinal ranges of the Qinghai-Tibetan Plateau (QTP). RESULTS: Independent phylogenetic relationships reconstructed using DNA sequences of the uniparentally inherited chloroplast genome and two low-copy nuclear genes confirmed the hypothesis of multiple origins of indehiscent sporangia and the restriction of particular haplotypes to indehiscent sporangia populations in the Lhasa and Nyingchi regions of the QTP. In contrast, the Hengduan Mountains were characterized by high haplotype diversity and the occurrence of accessions with and without indehiscent sporangia. Evidence was found for polyploidy and reticulate evolution in this complex. The putative case of chloroplast capture in the Nyingchi populations provided further evidence for the promotion of isolated but persistent populations by indehiscent sporangia. CONCLUSIONS: The presented results confirmed the hypothesis that indehiscent sporangia promote the establishment of persistent population in different regions of the QTP. These results are consistent with the expectations of reproductive reassurance by promotion of self-fertilization that played a critical role in the assembly of populations in isolated locations and/or extreme habitats.     

Comprehensive assessments of root biomass and production in a Kobresia humilis meadow on the Qinghai-Tibetan Plateau

Alpine meadow covers ca. 700,000 km² with an extreme altitude range from 3200 m to 5200 m. It is the most widely distributed vegetation on the vast Qinghai-Tibetan Plateau. Previous studies suggest that meadow ecosystems play the most important role in both uptake and storage of carbon in the plateau. The ecosystem has been considered currently as an active “CO₂ sink”, in which roots may contribute a very important part, because of the large root biomass, for storage and translocation of carbon to soil. To bridge the gap between the potential importance and few experimental data, root systems, root biomass, turnover rate, and net primary production were investigated in a Kobresia humilis meadow on the plateau during the growing season from May to September in 2008 and 2009. We hypothesized that BNPP/NPP of the alpine meadow would be more than 50%, and that small diameter roots sampled in ingrowth cores have a shorter lifespan than the lager diameter roots, moreover we expected that roots in surface soils would turn over more quickly than those in deeper soil layers. The mean root mass in the 0–20 cm soil layer, investigated by the sequential coring method, was 1995?±?479 g?m^?2 and 1595?±?254 g?m^?2 in growing season of 2008 and 2009, respectively. And the mean fine root biomass in ingrowth cores of the same soil layer was 119?±?37 g?m^?2 and 196?±?45 g?m^?2 in the 2 years. Annual total NPP was 12387 kg?ha^?1?year^?1, in which 53% was allocated to roots. In addition, fine roots accounted for 33% of belowground NPP and 18% of the total NPP, respectively. Root turnover rate was 0.52 year^?1 for bulk roots and 0.74 year^?1 for fine roots. Furthermore, roots turnover was faster in surface than in deeper soil layers. The results confirmed the important role of roots in carbon storage and turnover in the alpine meadow ecosystem. It also suggested the necessity of separating fine roots from the whole root system for a better understanding of root turnover rate and its response to environmental factors.     

Shiga Toxin-Producing Escherichia coli in Yaks (Bos grunniens) from the Qinghai-Tibetan Plateau,China

Shiga toxin (Stx)-producing Escherichia coli (STEC) are recognized as important human pathogens of public health concern. Many animals are the sources of STEC. In this study we determined the occurrence and characteristics of the STEC in yaks (Bos grunniens) from the Qinghai-Tibetan plateau, China. A total of 728 yak fecal samples was collected from June to August, 2012 and was screened for the presence of the stx₁ and stx₂ genes by TaqMan real-time PCR after the sample was enriched in modified Tryptone Soya Broth. Of the 138 (18.96%) stx₁ and/or stx₂-positive samples, 85 (61.59%) were confirmed to have at least 1 STEC isolate present by culture isolation, from which 128 STEC isolates were recovered. All STEC isolates were serotyped, genotyped by pulsed-field gel electrophoresis (PFGE) and characterized for the presence of 16 known virulence factors. Fifteen different O serogroups and 36 different O:H serotypes were identified in the 128 STEC isolates with 21 and 4 untypable for the O and H antigens respectively. One stx₁ subtype (stx_1a) and 5 stx₂ subtypes (stx_2a, stx_2b, stx_2c, stx_2d and stx_2g) were present in these STEC isolates. Apart from lpfA_O157/OI-141, lpfA_O157/OI-154, lpfA_O113, katP and toxB which were all absent, other virulence factors screened (eaeA, iha, efa1, saa, paa, cnf1, cnf2, astA, subA, exhA and espP) were variably present in the 128 STEC isolates. PFGE were successful for all except 5 isolates and separated them into 67 different PFGE patterns. For the 18 serotypes with 2 or more isolates, isolates of the same serotypes had the same or closely related PFGE patterns, demonstrating clonality of these serotypes. This study was the first report on occurrence and characteristics of STEC isolated from yaks (Bos grunniens) from the Qinghai-Tibetan plateau, China, and extended the genetic diversity and reservoir host range of STEC.     

环境变异对青稞热稳定蛋白质含量及蛋白质Z的影响

热稳定蛋白是衡量麦芽品质的重要指标,为探明青稞籽粒和麦芽热稳定蛋白的含量、蛋白质Z的组成特征以及影响条件。本研究以3份青稞和1份对照大麦品种Gairdner为试验材料,对青稞籽粒及其麦芽的热稳定蛋白进行分析与鉴定,研究了不同生态环境下青稞热稳定蛋白质含量和蛋白质Z的组成特征,同时筛选出了优异啤用品质青稞品种(品系)。结果表明,青稞发芽温度为20℃,发芽时间为72 h,培养溶液PH为5时,发芽及焙焦条件下最有利于青稞热稳定蛋白总含量及蛋白质Z的累积。利用该发芽条件筛选种植于西宁、湟源和海晏的青稞资源,发现种植于西宁的青稞种子和发芽后热稳定蛋白质总含量最低,但是焙焦后热稳定蛋白质和蛋白质Z含量最高;同时从150份青稞资源中筛选出热稳定蛋白质含量及蛋白质Z条带清晰、含量高的优异资源15份。本研究结果为酿造青稞品种选育、啤用青稞和麦芽质量评价指标提供理论依据。     

Flavobacterium qiangtangensis sp. nov., Isolated from Qiangtang Basin in Qinghai-Tibetan Plateau,China

A flexirubin-type yellow-pigmented, non-gliding, non-flagellated, gram-negative bacterium strain, designated F3^T, was isolated from a drilling core sample of the Qiangtang basin, Qinghai-Tibetan plateau, China. Phylogenetic analysis based on 16S rRNA gene sequences showed that the strain F3^T belongs to the genus Flavobacterium, with the highest 16S rRNA gene sequence similarity to the Flavobacterium noncentrifugens CGMCC 1.10076^T (94.92 %). Strain F3^T grew optimally at temperature about 20 °C, at pH about 7.0–8.0, at NaCl concentration 0 % (w/v). The DNA G+C content of the isolate was 35.5 mol%. The major polar lipid was phosphatidylethanolamine, predominant cellular fatty acids of the strain was iso-C_15:0 (22.02 %), while the major menaquinone was menaquinone 6. Due to the phenotypic and genetic distinctiveness and several other characteristic studied in this article, we consider F3^T as a novel species of the genus Flavobacterium, and propose to name it Flavobacterium qiangtangensis sp. nov. The type strain is F3^T (=CGMCC 1.12706^T = JCM 19739^T).     

青藏高原六种棘豆属植物的染色体数目及核型报道

对6种棘豆属植物（Oxytropis）的13个居群样进行细胞学研究,其中黄花棘豆（O．ochrocephala）、绢毛棘豆（O．tatarica）、甘肃棘豆（O．kansuensis）和铺地棘豆（0．humifusa）为首次报道染色体数目（2n=16）和核型;在胀果棘豆（O．stracheyana）中（2n=48）首次发现B染色体。现有的细胞学资料表明：棘豆属植物中多倍体占总报道数的58％,这说明多倍化在本属植物的进化过程中起着非常重要的作用,但青藏高原仅有一种植物发现多倍体,多倍化并不占主导地位,而主要表现为二倍体水平上的结构变异,即核型不对称性的变化。     

The USDA Barley Core Collection: Genetic Diversity,Population Structure,and Potential for Genome-Wide Association Studies

New sources of genetic diversity must be incorporated into plant breeding programs if they are to continue increasing grain yield and quality, and tolerance to abiotic and biotic stresses. Germplasm collections provide a source of genetic and phenotypic diversity, but characterization of these resources is required to increase their utility for breeding programs. We used a barley SNP iSelect platform with 7,842 SNPs to genotype 2,417 barley accessions sampled from the USDA National Small Grains Collection of 33,176 accessions. Most of the accessions in this core collection are categorized as landraces or cultivars/breeding lines and were obtained from more than 100 countries. Both STRUCTURE and principal component analysis identified five major subpopulations within the core collection, mainly differentiated by geographical origin and spike row number (an inflorescence architecture trait). Different patterns of linkage disequilibrium (LD) were found across the barley genome and many regions of high LD contained traits involved in domestication and breeding selection. The genotype data were used to define ‘mini-core’ sets of accessions capturing the majority of the allelic diversity present in the core collection. These ‘mini-core’ sets can be used for evaluating traits that are difficult or expensive to score. Genome-wide association studies (GWAS) of ‘hull cover’, ‘spike row number’, and ‘heading date’ demonstrate the utility of the core collection for locating genetic factors determining important phenotypes. The GWAS results were referenced to a new barley consensus map containing 5,665 SNPs. Our results demonstrate that GWAS and high-density SNP genotyping are effective tools for plant breeders interested in accessing genetic diversity in large germplasm collections.     

青藏高原东缘高寒草甸植物群落的开花物候

利用2008年整个生长季的地面花期物候观测数据,初步研究了青藏高原东缘高寒草甸植物群落开花物候的动态、格局、参数间的联系及其与物种共存的关系.结果表明:植物群落的花期从5月初开始,8月上旬进入开花数目的高峰期,到10月初结束,遍及整个生长季节;不同物种进入花期的时间、峰值期及持续时间不同,各物种花期物候生态位相分离,但群落中大部分物种开花峰值期主要集中在7-8月;不同物种的开花峰值期时间与花期持续时间有负相关关系:开花越早的植物其花期持续时间越长;物种间的开花物候差异主要是由植物本身的特性和环境条件共同决定的,而群落水平上的开花物候格局主要受非生物因素的控制.

Abstract：

Based on the observation on the flowering phenology in the whole growth season of 2008, the dynamics, patterns, and relationships among various parameters of flowering phenology of alpine meadow plant community in eastern Qinghai-Tibetan Plateau were studied, and the potential effects of above-mentioned factors on the species coexistence of the plant community were approached. The flowering period of the plant community started from early May, came into peak period in the first ten days of August, and ended in early October, extending all over whole growth period. Different species had their different first flowering date, peak flowering period, and flowering duration, and their flowering phenologieal niches differentiated obviously. Most species had their peak flowering period centralized in July-August. There was a negative rela-tionship between peak flowering period and flowering duration: the earlier the flowering, the lon-ger the flowering duration. The differences in flowering phonology among the species were mainly determined by species per se characteristics and environmental conditions, but the flowering phe-nological pattern on community level was primarily controlled by abiotic factors.