The oxygen and carbon dioxide compensation points of C3 plants: possible role in regulating atmospheric oxygen.

The O2 and CO2 compensation points (O2 and CO2) of plants in a closed system depend on the ratio of CO2 and O2 concentrations in air and in the chloroplast and the specificities of ribulose bisphosphate carboxylase/oxygenase (Rubisco). The photosynthetic O2 is defined as the atmospheric O2 level, with a given CO2 level and temperature, at which net O2 exchange is zero. In experiments with C3 plants, the O2 with 220 ppm CO2 is 23% O2; O2 increases to 27% with 350 ppm CO2 and to 35% O2 with 700 ppm CO2. At O2 levels below the O2, CO2 uptake and reduction are accompanied by net O2 evolution. At O2 levels above the O2, net O2 uptake occurs with a reduced rate of CO2 fixation, more carbohydrates are oxidized by photorespiration to products of the C2 oxidative photosynthetic carbon cycle, and plants senesce prematurely. The CO2 increases from 50 ppm CO2 with 21% O2 to 220 ppm with 100% O2. At a low CO2/high O2 ratio that inhibits the carboxylase activity of Rubisco, much malate accumulates, which suggests that the oxygen-insensitive phosphoenolpyruvate carboxylase becomes a significant component of the lower CO2 fixation rate. Because of low global levels of CO2 and a Rubisco specificity that favors the carboxylase activity, relatively rapid changes in the atmospheric CO2 level should control the permissive O2 that could lead to slow changes in the immense O2 pool.

Oxygen Requirement and Inhibition of C4 Photosynthesis1 : An Analysis of C4 Plants Deficient in the C3 and C4 Cycles

The basis for O2 sensitivity of C4 photosynthesis was evaluated using a C4-cycle-limited mutant of Amaranthus edulis (a phosphoenolpyruvate carboxylase-deficient mutant), and a C3-cycle-limited transformant of Flaveria bidentis (an antisense ribulose-1,5-bisphosphate carboxylase/oxygenase [Rubisco] small subunit transformant). Data obtained with the C4-cycle-limited mutant showed that atmospheric levels of O2 (20 kPa) caused increased inhibition of photosynthesis as a result of higher levels of photorespiration. The optimal O2 partial pressure for photosynthesis was reduced from approximately 5 kPa O2 to 1 to 2 kPa O2, becoming similar to that of C3 plants. Therefore, the higher O2 requirement for optimal C4 photosynthesis is specifically associated with the C4 function. With the Rubisco-limited F. bidentis, there was less inhibition of photosynthesis by supraoptimal levels of O2 than in the wild type. When CO2 fixation by Rubisco is limited, an increase in the CO2 concentration in bundle-sheath cells via the C4 cycle may further reduce the oxygenase activity of Rubisco and decrease the inhibition of photosynthesis by high partial pressures of O2 while increasing CO2 leakage and overcycling of the C4 pathway. These results indicate that in C4 plants the investment in the C3 and C4 cycles must be balanced for maximum efficiency.

n-alkane carbon isotope ratios in plants and recent lake sediments from Cameroon

Trees and shrubs in tropical Africa use the C3 cycle as a carbon fixation pathway during photosynthesis, while grasses and sedges mostly use the C4 cycle. Leaf-wax lipids from sedimentary archives such as the long-chain n-alkanes (e.g., n-C27 to n-C33) inherit carbon isotope ratios that are representative of the carbon fixation pathway. Therefore, n-alkane d13C values are often used to reconstruct past C3/C4 composition of vegetation, assuming that the relative proportions of C3 and C4 leaf waxes reflect the relative proportions of C3 and C4 plants. We have compared the d13C values of n-alkanes from modern C3 and C4 plants with previously published values from recent lake sediments and provide a framework for estimating the fractional contribution (areal-based) of C3 vegetation cover (fC3) represented by these sedimentary archives. Samples were collected in Cameroon, across a latitudinal transect that accommodates a wide range of climate zones and vegetation types, as reflected in the progressive northward replacement of C3-dominated rain forest by C4-dominated savanna. The C3 plants analysed were characterised by substantially higher abundances of n-C29 alkanes and by substantially lower abundances of n-C33 alkanes than the C4 plants. Furthermore, the sedimentary d13C values of n-C29 and n-C31 alkanes from recent lake sediments in Cameroon (-37.4 per mil to -26.5 per mil) were generally within the range of d13C values for C3 plants, even when from sites where C4 plants dominated the catchment vegetation. In such cases simple linear mixing models fail to accurately reconstruct the relative proportions of C3 and C4 vegetation cover when using the d13C values of sedimentary n-alkanes, overestimating the proportion of C3 vegetation, likely as a consequence of the differences in plant wax production, preservation, transport, and/or deposition between C3 and C4 plants. We therefore tested a set of non-linear binary mixing models using d13C values from both C3 and C4 vegetation as end-members. The non-linear models included a sigmoid function (sine-squared) that describes small variations in the fC3 values as the minimum and maximum d13C values are approached, and a hyperbolic function that takes into account the differences between C3 and C4 plants discussed above. Model fitting and the estimation of uncertainties were completed using the Monte Carlo algorithm and can be improved by future data addition. Models that provided the best fit with the observed d13C values of sedimentary n-alkanes were either hyperbolic functions or a combination of hyperbolic and sine-squared functions. Such non-linear models may be used to convert d13C measurements on sedimentary n-alkanes directly into reconstructions of C3 vegetation cover.

Late Quaternary vegetational and climatic changes from tropical peats in southern India - An extended record up to 40,000 years BP

Stable carbon isotope ratios of peats dated (by C-14) back to 40 kyr BP from the montane region (> 1800 m asl) of the Nilgiris, southern India, reflect changes in the relative proportions of C3 and C4 plant types, which are influenced by soil moisture (and hence monsoonal precipitation), From prior to 40 kyr BP until 28 kyr BP, a general decline in delta(13)C values from about - 14 per mil to - 19 per mil suggests increased dominance of C3 plants concurrent with increasingly moist conditions, During 28-18 kyr BP there seems relatively little change with delta(13) C of - 19 to - 18 per mil, At about 16 kyr BP a sharp reversal in delta(13)C to a peak of - 14.7 per mil indicates a clear predominance of C4 vegetation associated with arid conditions, possibly during or just after the Last Glacial Maximum, A moist phase at about 9 kyr BP (the Holocene Optimum) with dominance of C3 vegetation type is observed, while arid conditions are re-established during 5-2 kyr BP with an overall dominance of C4 vegetation, New data do not support the occurrence of a moist phase coinciding with the Mediaeval Warm Period (at 0.6 kyr BP) as suggested earlier, Overall, the climate and vegetation in the high altitude regions of the southern Indian tropics seem to have responded to past global climatic changes, and this is consistent with other evidences from India and other tropical regions.

Stable carbon isotope ratios in Asian elephant collagen: implications for dietary studies

Stable carbon isotope ratios in bone collagen have been used in a variety of dietary studies in modern and fossil animals, including humans. Inherent in the stable isotope technique is the assumption that the isotopic signature is a reflection of the diet and is persistent in collagen because this is a relatively inert protein. Carbon isotope analyses of bones from a southern Indian population of Asian elephant (Elephas maximus), a long-lived mammal that alternates seasonally between a predominantly C3 (browse) and C4 (grass) plant diet, showed two patterns that have important implications for dietary interpretation based on isotopic studies. Relative to the quantity of the two plant types consumed on average, the ?13C signal in collagen indicated that more carbon was incorporated from C3 plants, possibly due to their higher protein contribution. There was a much greater variance in ?13C values of collagen in sub-adult (range -10.5� to-22.7�, variance=14.51) compared to adult animals (range -16.0� to -20.3�, variance=1.85) pointing to high collagen turnover rates and non-persistent isotopic signatures in younger, growing animals. It thus seems important to correct for any significant relative differences in nutritive value of food types and also consider the age of an animal before drawing definite conclusions about its diet from isotope ratios.

谷子和甘蔗C4型磷酸烯醇式丙酮酸羧化酶基因的克隆及其转基因水稻光合效率的提高

水稻是我国重要的粮食作物之一，它是一种典型的C3植物。与其它C3作物不一样的是，水稻的生长需要相对较高的温度和充足的阳光照射。然而高温和高光强的生长环境更加适合于C4植物的生长，更加有利于发挥C4植物高光合效率的特点。因此本论文希望将C4植物中固定CO2的酶磷酸烯醇式丙酮酸羧化酶基因导入水稻，获得一种更加适合高温和高光强生活环境的“C4型”水稻，这对于提高水稻的产量，满足人口增长对粮食需求具有重大意义。 本论文从C4植物谷子和甘蔗中克隆了其C4型磷酸烯醇式丙酮酸羧化酶cDNA基因，获得了具有自主知识产权的基因克隆，并将它们导入粳稻品种中花8号，进而对转基因材料的光合生理特性进行了研究。结果如下： 首次从谷子中得到了ppc基因两个cDNA克隆，分别命名为Mppc1和Mppc2。前者是一个C3型的ppc基因，它可能属于在根中特异表达的C3-2型ppc基因;后者是在绿色叶片中大量表达的C4型ppc基因。它们所编码的蛋白的氨基酸残基数分别为961和964，序列同源性为82.5%。C4型PEPC多出的3个氨基酸位于N末端。利用RACE的方法我们得到了谷子C4型ppc基因完整的cDNA序列，包括63bp的5'非编码区，2895bp的编码区和256bp的3'非编码区。 首次获得了甘蔗C4型ppc基因完整的cDNA序列的克隆，命名为Sppc。它包括95bp的5'非编码区、2886bp的编码区，和224bp的3'非编码区。 利用所克隆的基因，分别连上强组成型启动子Ubiquitin启动子和强光调控启动子Rubisco小亚基启动子后，再插入两个标记基因不同的表达载体pCB和pPCB的多克隆位点中，构建了八个含有外源ppc基因的植物表达载体pCB-Pubi-Mppc、pCB-Pubi-Sppc、pCB-PrbcS-Mppc、pCB-PrbcS-Sppc、pPCB-Pubi-Mppc、pPCB-Pubi-Sppc、pPCB-PrbcS-Mppc和pPCB-PrbcS-Sppc。再加上含有玉米完整的C4型ppc 核基因的载体pCB-ZMppc，共有9个载体。利用农杆菌介导法进行了水稻的转化，各个载体都获得了大量的转基因植株。对标记基因潮霉素磷酸转移酶基因hpt和磷酸甘露糖异构酶基因pmi以及导入的目的ppc基因的PCR扩增检测，结果显示绝大多数转基因植株都能扩增出目的片段，而未转化的植株则没有扩增产物。对部分转基因水稻的Southern和Western杂交以及RT-PCR分析都表明，无论从DNA水平、mRNA水平，还是从蛋白质水平上都证明外源ppc基因都成功地导入了水稻，并获得了正确的表达。 对各载体转基因植株PEPC活性大规模的测定表明，转入玉米完整C4型PEPC核基因（有内含子）的水稻表现出极大的表达效率，大多数转基因材料的PEPC活性为对照的10-20倍，其活性最高可达到对照的44倍。转入谷子和甘蔗PEPC基因cDNA的水稻，表达的效率很低，多数材料活性增加仅为对照的2-5倍，但也有极少数材料活性增加了10倍以上。用Rubisco小亚基启动子控制的ppc基因在水稻的表达活性要略高于Ubiquitin启动子控制的ppc基因。以上结果说明ppc基因的内含子在其转录或mRNA的稳定上起着重要作用。 对部分转基因材料气体交换特征的研究发现，随着转基因水稻PEPC活性的增加，净光合速率也有逐渐增加的趋势。其中PEPC活性最大的ZM24株系的三个单株净光合速率比对照增加了39.8%、13.7%和28.6%，而它们的PEPC活性比对照分别增加了21.2、21.9和23.6倍。 转PEPC水稻的净光合速率与气孔导度具有显著的相关性。这说明表达的外源ppc 基因产物PEPC参与了转基因水稻的气孔运动，使气孔开放程度增加。更有意义的是过表达PEPC的水稻具有更高的水分利用效率，这就增加了其耐旱能力。在光抑制条件下转基因水稻也具有更高的光合能力。这些特征表明转ppc基因的水稻比对照更加适合于水稻高温高光强和干旱的原生环境。

CO2浓度升高对红桦幼苗生理与生长的影响

大气CO2浓度的增加已经成为不可争议的事实。预计本世纪末大气CO2浓度将增加到约700µmol mol-1。森林年光合产量约占陆地生态系统年光合产量的70%。森林树木是一个巨大的生物碳库，约占全球陆地生物碳库的85%。森林树木对CO2的固定潜力是缓解由大气CO2浓度升高引起的未来全球气候变化问题的决定性因子之一。红桦（Betula albosinensis Burk.）是川西亚高山采伐迹地自然或人工恢复的重要树种。本研究以1a红桦幼苗为模式植物，采用人工模拟的方法，研究CO2浓度升高对不同种内竞争强度（种群水平）下红桦幼苗的生理特征、生长、干物质积累及其分配的影响，探讨在种内竞争生长条件下红桦幼苗的“光合适应机理”与生长特征，为西南亚高山森林生产力对未来全球变化的预测提供重要参考。 本研究的主要结果如下： 1）在种内竞争生长条件下红桦幼苗经过CO2浓度升高熏蒸4个月后，叶片出现“光合适应”现象。与对照相比，低种植密度（28株m-2）和高种植密度（84株m-2）条件下的红桦幼苗净光合速率（A）、气孔导度（gs）、蒸腾速率（E）、表观量子产量（AQY）和羧化速率（CE）显著降低，而水分利用效率（WUE）则显著提高。CO2浓度升高处理的红桦幼苗叶片Rubisco活性、单位叶面积N浓度、叶绿素a、叶绿素b和类胡萝卜素浓度都显著降低。但CO2浓度对红桦幼苗的叶绿素a与叶绿素b的比值没有显著影响。CO2浓度升高显著增加红桦幼苗单位叶面积的非结构性碳水化合物（TNC）浓度，结果是红桦幼苗的比叶面积（SLA，cm2 g-1）显著降低。 2）与对照相比，CO2浓度升高处理的红桦幼苗高、基径、单叶面积和侧枝的相对生长速率（R GR）显著提高，尤其在试验处理的早期。CO2浓度升高既增加单株红桦幼苗总叶片数量又增加单叶面积，结果是单株红桦幼苗的总叶面积比对照显著增加。 3）CO2浓度升高处理显著增加红桦幼苗干物质积累（尤其是细根生物量），改变了红桦幼苗生物量的分配格局。与对照相比，CO2浓度升高处理的红桦幼苗叶重比（LWR）、叶面积比（LAR）、叶根重比（Wl/Wr）和源汇重比（leaf weight to non-leaf weight ratio, Wsource/Wsink）显著下降（高种植密度的LWR除外），而根冠比（R/S）则显著增加。在两种种植密度条件下，CO2浓度升高显著增加红桦幼苗根生物量的分配比率，显著降低叶片的生物量分配比率，对主茎、侧枝以及地上生物量的分配比率不变或约有下降。 总之，长期生长在CO2浓度升高条件下的红桦幼苗光合能力下降，并伴随Rubisco活性、叶N浓度、光合色素浓度的显著降低以及TNC浓度的显著增加。支持树木光合速率下降与Rubisco活性、叶N浓度下降以及TNC浓度增加紧密相关的假设。CO2浓度升高处理红桦幼苗的早期相对生长速率大大高于对照，而后期迅速下降，说明红桦幼苗生物量的显著增加主要归功于CO2浓度升高的早期促进作用和叶面积的显著增加。CO2浓度升高显著增加红桦幼苗根系生物量和根冠比，表明红桦幼苗“额外”固定的C向根系转移。 The steady increae of atmospheric CO2 concentration（[CO2]）has been inevitable fact. Models predict that the atmospheric [CO2] will increase to about 700µmol mol-1 at the end of the twenty-first century. As trees constitute a majoor carbon reservoir–85% of total plant carbon is found in forest, and their ability to sequester carbon is a key determinant of future global change problems caused by increases in atmospheric CO2. In addition to the role of forests in the global carbon cycle, inceased growth could be of economic benefit, for example, offsetting deleterious effects of climatic changes. Betula albosinensis (Burk.) usually emerges as the pioneer species in initial stage and as constructive species in later stages of forest community succession of mountain forest area, and also is one of important tree species for afforestation in logged area, in southwesten China. In this experinment, Betula albosinensis seedling (one-year-old) was used as the model plant. B. albosinensis seedlings were grown under two all-day [CO2], ambient (about 350 µmol·mol-1) and elevated [CO2] (about 700 µmol·mol-1), and two planting densities of 28 plants per m2 and 84 plants per m2. The objectives were to characterize birch mature leaf photosynthesis, growth, mass accumulation and allocation responses to long-tern elevated growth [CO2] under the influences of neighbouring plants, and to assess whether elevated [CO2] regulated birch mature leaf photosynthetic capacity, in terms of leaf nitrogen concentration (leaf [N]), activity of ribulose bisphosphate carboxygenase (Rubisco), Rubisco photosynthetic efficiency, and total nonstructural carbohydrates (TNC) concentration, and also to provide a strong reference to predict the productivity of subalpine forests under the future global changes. The results are as follows: 1) B.albosinensis seedlings exposed to elevated [CO2] for 120 days, photosynthetic acclimation phenomena occurred. At two planting densities, leaves of birch seedlings grown under elevated [CO2] had lower net photosynthetic rate (A), stomatal conductance (gs), transpiration (E), apparent quantum yield (AQY) and carboxylated efficiency (CE) and higher water use efficiency (WUE), compared to those of B.albosinensis seedlings grown under ambient [CO2]. Based on the leaf area, leaf [N], Rubisco activity and photosynthetic pigments concentrations of B. albosinensis seedlings grown under elevated [CO2] were significantly lower than those grown under ambient [CO2]. The ratio of chlorophyll a to chlorophyll b concentration was not affected by elevated [CO2]. Under elevated [CO2], the TNC concentration per unit leaf area significantly increased, resulting in significant decrease in specific leaf area. Thus leaf photosynthetic capacity of B. albosinensis seedlings would perform worse under rising atmospheric [CO2] and the influences of neighbouring plants. 2) Under elevated [CO2], the relative growth rate (RGR) of B. albosinensis seedlings height, basal diameter, a leaf area and branch length significantly increased, especially at the initial stage of exposure to elevated [CO2], and a leaf area and leaf numbers per B. albosinensis seedling also significantly increased. Thus the total leaf area per B. albosinensis seedling was significantly increased under elevated [CO2]. 3) As the increase of RGR and total leaf area, biomass of B. albosinensis seedling grown elevated [CO2] was higher, compared to that of B.albosinensis seedlings grown at ambient [CO2]. Elevated [CO2] changed the biomass allocation pattern of B. albosinensis seedling. At two planting densities, B. albosinensis seedlings grown elevated [CO2] had lower leaf weight to total weight ratio (LWR), leaf area to total weight ratio (LAR) and leaf weight to non-leaf weight ratio (Wsource/Wsink), but higher root weight to shoot weight ratio (R/S), compared to those of B.albosinensis seedlings grown at ambient [CO2]. Under elevated [CO2], roots biomass to total biomass ratio was signigicantly increased, leaves biomass to total biomass ratio was significantly decreased. The main stem and branch biomass to total biomass ratio were not affected by elevated [CO2]. In conclusion, our results supported the hypothesis that the decline in photosynthetic capacity of C3 plants will appear after long-term exposure to elevated [CO2], accompanying with the significant decrease in Rubisco activity, leaf N concentration, photosynthetic pigments concentration, and significant increase in total non-structural carbohydrates concentration. Our results also have shown that the increase of biomass of B. albosinensis seedlings should be attributed to initial stimulation on RGR and total leaf area resulted from elevated [CO2]. Under elevated [CO2], the extra carbon sequestered by B.albosinensis seedlings transferred into under-ground part because of increase in root biomass and R/S.

水稻同源四倍体三系法杂种优势利用研究

水稻是重要的粮食作物，其产量的增加和品质的改良都是关系国计民生的大事。就我国现阶段的国情而言，水稻产量在现有水平上稳步提升仍是未来十几年甚至几十年农业生产最重要的目标之一。尽管根据“超级杂交水稻育种”的战略设想和水稻育种实践，通过不断地改进育种技术可望在更高的产量水平上进行水稻杂种优势利用，在稻属植物内还具有很大的产量潜力可以挖掘。然而，仅仅从现有的种质基础出发，要更大幅度提高水稻单产，实现“超级杂交稻”的目标也存在一些困难：现有的推广品种是二倍体，尽管种类众多，但是其基因组的来源相对单一；同时，水稻基因组DNA含量也是作物中最少的，基因组内寻求开发潜力有一定困难；水稻作为C3植物，光合利用效率不高也是制约水稻产量提高的因素之一。因此，寻求常规手段以外的技术突破或者方法创新，是实现“超级杂交稻”的目标的迫切需求。本研究利用秋水仙素能抑制细胞分裂中纺锤丝的收缩、使细胞染色体加倍的作用，对水稻幼穗诱导的愈伤组织细胞进行加倍，并分化出再生植株；创制出水稻同源四倍体新的种质材料，在此基础上选育水稻同源四倍体雄性不育三系材料，并实现水稻同源四倍体的三系配套，开展水稻同源四倍体杂种优势利用和四倍体杂交水稻选育研究，建立水稻同源四倍体杂种优势利用的新技术体系。这不仅有助于倍性水平杂种优势的开拓和利用，同时也将为我国新世纪“超级稻”育种研究开辟一条新的技术途径。 水稻幼穗诱导愈伤组织并分化成苗是一项成熟、简单的组织培养技术。本研究以普通二倍体水稻亲本为材料，用秋水仙素进行水稻的多倍体化诱导，创制同源四倍体水稻三系亲本材料并对其进行鉴定。多倍体化以秋水仙素诱导的愈伤组织培养为基础，研究不同秋水仙素浓度梯度和愈伤组织诱导培养基组合对诱导四倍体植株的影响。结果表明在MS+2,4 D 1.0mg/L+ KT0.2mg/L+ IAA0.2mg/L 和500mg/L的秋水仙素处理下，水稻愈伤组织染色体加倍（有最高的效率）效果较好，平均加倍频率可达25.26%，其中，材料CDR22和IR26诱导较易成功，加倍频率分别达到75%和26.5%；相对材料94109 1.3%加倍频率和冈46B 10.8%加倍频率，诱导率差异极显著。 对水稻四倍体材料进行了形态学鉴定结果表明，与二倍体水稻对照相比其株高、穗长、花粉育性等主要农艺性状，确定四倍体材料在穗长和千粒重两方面极显著提高，种子的长度和宽度也显著增长。对花粉育性鉴定，确认水稻四倍体不育系材料仍为不育，保持系材料自交和杂交可育，恢复系材料自交和杂交可育。对四倍体材料进行细胞形态、染色体数目等方面进行细胞学鉴定，经核型分析表明水稻四倍体材料具有48条染色体，是二倍体水稻的两倍。水稻四倍体材料根尖分生组织细胞与二倍体的根尖分生组织细胞相比，细胞体积、细胞核和核仁显著增大。四倍体三系材料在细胞有丝分裂中期均可规则排列在赤道板，并能均等地移向两极；后期观察中没有发现染色体分离滞后现象，分裂末期细胞能够形成大小相对均一的子细胞。水稻同源四倍体三系材料细胞分裂未见异常，植株生长发育正常。 从1996年至2006年，针对结实率、有效分蘖、着粒数和穗长等主要农艺性状，通过系谱选育的方法，对培育的同源四倍体水稻亲本材料进行了连续选择和改良，取得较好成效。表现为结实率的改良效果极佳，所有改良材料的平均结实率均呈上升趋势，如D237（29.70％→72.70％）、DTB（19.55％→53.21％）等。有效分蘖总体呈现上升趋势，但在不同的年份，如1998和2002存在较大的负向波动。部分材料改良效果明显，如D19B（5.87→13.50）、D什香 (7.00→12.00)等；同时一些材料如DTB和D明恢63虽然总体略有提高，但在不同的年份波动很大，因此存在较大改良阻力，原因还有待进一步研究。着粒数的改良上升趋势比较显著，除保持系的DTB之外，其余材料的平均着粒数有显著提高。穗长的改良阻力较大，虽然不同材料总体上有所提高，但效果并不显著，并且不同年份有较大负向波动（2001）。此外还对株高、剑叶长等性状也进行了选择，但效果不显著，原因有待进一步提高。同源四倍体材料产量相关性状遗传改良幅度不一致，保持系和恢复系间的遗传改良效果也存在差异。这为同源四倍体水稻的进一步利用打下了良好的基础。 籼稻和粳稻亚种间杂交及杂种优势利用的主要障碍就是其低的结实率。而同源四倍体杂交水稻的研究为提高杂交水稻的杂种优势利用创造了新的途径。本研究通过随机区组设计方案，挑选性状优良的二倍体水稻材料，包括雄性不育系，保持系和恢复系进行秋水仙素诱导加倍，从而获得同源四倍体水稻对应的三系材料。利用选育的优良水稻同源四倍体三系材料，配制7个杂交组合，杂交F1代与其恢复系亲本进行比较，用于计算超亲优势（HB）；而杂交F1代与生产上大面积推广的二倍体杂交品种汕优63进行比较，用于计算杂种优势。结果显示，同源四倍体杂交水稻的超亲优势表现为：每株有效穗变化幅度为1.4%至105.9%，总粒数为0.5%至74.3%，每穗实粒数为17.6%至255.7%，结实率为9.6%至130.4%。这些农艺性状的改良使得这7个杂种F1的理论产量的超亲优势高达64.8%至672.7%。小区试验中四倍体杂交水稻组合T461A/T4002和T461A/T4193分别比二倍体对照汕优63提高46.3%和38.3%以上，除一个品种以外所有品种产量均接近或高于汕优63的产量。同源四倍体水稻强大的杂种优势表明，亚种间杂交育性低的问题可通过四倍体化及强化选择来解决。此外，同源四倍体杂交水稻器官的巨大性也是其产量提高的有利因素，水稻同源四倍体三系杂种优势利用研究具有一定的理论价值和商业生产潜力。 Rice is one of the major food crops, the improvement of the production and quality of it is an important thing related to the people's livelihood. On China's current national conditions, steadily increase of the rice yield based on the current level is still one of the most important goals in the next decade or even decades of agricultural production. According to the "super hybrid rice breeding" the strategic and rice breeding practice, improvement of the use of hybrid rice heterosis through continuous improvements in breeding technology is expected to get a higher level of rice yield, there are also a great yield potential can be exploited. However, there are also some difficulties to increase rice yield obviously and implement the goal of "super hybrid rice" based on the existing germplasm: Rice varieties in promotion are diploid, although there are many varieties, but their genome are from a comparatively single source; Meanwhile, the rice genome DNA are the least among the crops, it is difficult to exploit the development potential within the genome; Rice as C3 plants, photosynthetic efficiency is not high, it is one of the factors constraint rice yield. Therefore, seeking technological breakthroughs or innovative methods different from conventional means is the urgent needs to reach the target of "super hybrid rice". Using colchicine inhibit spindle contraction during cell division, double the cell chromosome, we induced callus cells from rice panicle to be doubled, and differentiated regeneration; we created a new autotetraploid rice germplasm material, and on that basis we bred male sterility three line autotetraploid rice materials, and the achieved the three line rice autotetraploid matchmaking, researched in autotetraploid rice heterosis usage and tetraploid hybrid rice breeding, constituted a new technology system of autotetraploid hybrid rice heterosis utilization. This not only helps the tetraploid rice heterosis exploration and use, but also inaugurates a new technical means for China in the new century "super rice" breeding research. We chose ordinary diploid rice as materials, using colchicine to induce the polyploidization, created the autotetraploid rice three-line materials and identified them. The polyploidization was based on the colchicine-induced callus tissue culture, and we experimented different colchicine concentrations and culture mediums to induce tetraploid plants, confirmed that the optimal concentration for inducement was 500 mg/L, the average induce rate was 25.26 %. Among all the materials, CDR22 and IR26 had higher induced rate; in contrary, 94109 and GANG46B had lower induced rate, the difference was significant. Autotetraploid materials was identified of both morphological and cytological, compared plant height, length of pollen sterility, and other major agronomic traits with a diploid rice as the control plant, identified that the autotetraploid materials had very significant advantages in ear length and thousand-grain weight, as well as the size of the seeds. Cytology identification included observation of the cell morphology, the number of chromosomes, and karyotype analysis on the autotetraploid materials confirmed that their chromosome number was 48, twice of the diploid rice. Mitoses in the three lines were common: chromosomes arrayed normally in metaphase and separated balanced into the two poles, chromosome moved without lagging in anaphase and daughter cells normally formed in telophase except one. It has been proved that tetraploid rice has normal meiosis as their diploid relatives, which usually including series of sub-phases as interphase, prophase I (five sub-phases), prophase II, metaphase I, II, anaphase I, II and telophase I, II. However, abnormal phenomena, such as formation of tetravalent, trivalent and univalent, chromosome lagging and so on, which would finally block meiosis. Configurations of chromosome in metaphaseⅠwere versatile in structure and form accept the bivalent. That condition varied in different strain, suggesting more complex paring configurations and more versatile genetic characters in tetraploid rice. All these abnormalities in meiosis contributed to low fertility of gamete and might consequently resulted in low seed setting. Successive selection and improvement on seed set, productive tiller per plant, total grains per panicle, panicle length and so on had been carried out from 1996 to 2006. The raise of seed sets was significant in both restorers and maintainers. Seed sets of some strains were improved more significantly than others, for example D237（29.70％→72.70％）、DTB（19.55％→53.21％）and et al.. Productive tiller per plant was improved to some extant. The tendency of improvement was rising on the whole but changed in some years such as 1998 and 2002. Part of the stains increased greatly, such as D19B（5.87→13.50）、Dshixiang (7.00→12.00) and so on, but some strains including DTB and Dminghui63 only increased little and decreased in some years by unknown reason. Total grains per panicle increased significantly and all strains except DTB increased. Improvement of panicle length termed to be hard. Different strains showed different capacities for improvement and floating existed in different years for example 2001. It has been proved that other agronomical traits including plant length, flag leaf length and so on could be improved but not significantly by selection also. In a word, agronomical traits could be raised by successive selection that is prerequisite for further utility of autotetraploid rice. Poor fertility is the main barrier for utilizing heterosis between the two rice (Oryza stiva L.) sub-species, indica and japonica. Recently, the development of autotetraploid hybrids (2n=4x=48) has been suggested as a new method for increasing heterosis in hybrid rice. Using standard experimental protocols, the elite diploid rice male sterile, maintainer, and restorer lines were colchine-doubled and autotetraploid counterparts were obtained. Seven resulting hybrids were analyzed for heterobeltiosis (HB), where the F1 was compared to the male parent, and the degree of heterosis, where the F1 was compared to the diploid commercial hybrid, Shanyou 63. The HB among the autotetraploid hybrids ranged from 1.4 to 105.9% for the productive panicles per plant, 0.5 to 74.3% for total kernels per panicle, 17.6 to 255.7% for filled kernels per panicle, and 9.6 to 130.4% for seed set. Improvements in these yield components resulted in the HB for kernel yield ranging from 64.8 to 672.7% among the seven hybrids. Hybrids T461A/T4002 and T461A/T4193 yielded 46.3 and 38.3% more, respectively than Shanyou 63, and all other hybrids but one yielded the same or more than Shanyou 63. The high heterosis for yield suggests that hybrid sterility between two rice sub-species may be overcome by using tetraploid lines followed by intensive selection. Also, the gigantic features of the autotetraploid hybrids may establish a plant structure able to support the higher yield.

碳、氧稳定同位素分析在古环境研究中的应用

A number of proxy records of paleoenvironment using stable isotopes could show the history of past environmental changes. These archives include peat and lake sediments, loess-paleosot sequence, fossil mammals and stalagmite, and so on. The stable isotopic composition of carbonate and organic matter and frequency magnetic susceptibility from Tianshuigou and Yuanlei loess-palesol sequence can be used to give estimates of the paleoenvironmental history of Dali, and even of the whole Chinese Loess Plateau during the last 250ka. Features of the High Temperature and Large Precipitation Event in the Tibet Plateau and its adjacent area during 40~30kaBP had been studied by Professor Shi Y. In this dissertation, its impact on Chinese Loess Plateau has been discussed again. Carbon and oxygen isotopic ratios, magnetic susceptibility and frequency magnetic susceptibility in Tianshuigou and Yuanlei profiles show that the Event in this area is not so stronger as the Tibet Plateau. The carbon isotopic composition of organic matter in Tianshuigou, Yuanlei, dingcun and Jingcun loess-palesol sequences are indicative of major changes in the paleovagetation between terrace and plain of the Chinese Loess Plateau. Water is one of the most important factors adjusting the relative biomass of C4 plant in terrestrial ecosystems. Stable carbon isotope ratio of vertebrate tooth enamel is used increasingly to reconstruct environmental and ecological information modern and ancient ecosystems. The SI3C value of tooth enamel bioapatites can distinguish between browsers and grazers. Data from typical grassland of Inter Mongolia, the Alpine meadow of Qinghai-Tibet Plateau and the Yaluzangbu Great Canyon indicate that diets of mammals could record the relative biomass of C4 plant only in the C4 dominated ecosystem. In a C3 dominated ecosystem, diet of mammals would include more C3 plants than vegetation. According to Professor Cerling, proxy records from North and South America, Africa and Pakistan show that at the end of the Miocene (between 8Ma to 6 Ma) there was a global expansion of CA biomass, probably when atmospheric CO2 levels declined. Thus, "C4 world" and "CO2 starvation" are put forward. In this dissertation, carbon isotopes of fossil tooth such as Equus sanmeniensis and Hipparion chiai from Linxia, China reveal that there is a C3 dominated ecosystem in the late Miocene. Diets of ancient mammals in Linxia are not evidence of global expansion of C4 biomass.

Detection of testosterone administration based on the carbon isotope ratio profiling of endogenous steroids: international reference populations of professional soccer players.

BACKGROUND AND OBJECTIVES: The determination of the carbon isotope ratio in androgen metabolites has been previously shown to be a reliable, direct method to detect testosterone misuse in the context of antidoping testing. Here, the variability in the 13C/12C ratios in urinary steroids in a widely heterogeneous cohort of professional soccer players residing in different countries (Argentina, Italy, Japan, South Africa, Switzerland and Uganda) is examined. METHODS: Carbon isotope ratios of selected androgens in urine specimens were determined using gas chromatography/combustion/isotope ratio mass spectrometry (GC-C-IRMS). RESULTS: Urinary steroids in Italian and Swiss populations were found to be enriched in 13C relative to other groups, reflecting higher consumption of C3 plants in these two countries. Importantly, detection criteria based on the difference in the carbon isotope ratio of androsterone and pregnanediol for each population were found to be well below the established threshold value for positive cases. CONCLUSIONS: The results obtained with the tested diet groups highlight the importance of adapting the criteria if one wishes to increase the sensitivity of exogenous testosterone detection. In addition, confirmatory tests might be rendered more efficient by combining isotope ratio mass spectrometry with refined interpretation criteria for positivity and subject-based profiling of steroids.

Immigration et alimentation à Montréal aux XVIIe et XVIIIe siècles : essai d’interprétation à partir d’analyses isotopiques sur des populations archéologiques

Afin d’étudier l’influence de la migration sur l’alimentation à Montréal aux XVIIe et XVIIIe siècles, 64 individus de la collection du cimetière Notre-Dame, daté de 1691 à 1796, ont fait l’objet d’analyses ostéologiques et isotopiques. Les analyses isotopiques ont portées sur le carbone (d13C) et l’azote (d15N) du collagène des os, ainsi que sur le d13C et l’oxygène (d18O) du carbonate de l’apatite des os et des dents (prémolaires et troisièmes molaires). Le d18O des dents a permis de définir approximativement trois régions d’origine (région de Montréal, région enrichie en 18O (i.e. Acadie, Louisiane, Nouvelle-Angleterre, France, Antilles et Afrique) et région appauvrie en 18O (intérieur des terres et plus au nord) pour 58 individus, et sept possibles parcours migratoire (N=27). Plus de la moitié de l’échantillon est composé d’individus possiblement natifs de Montréal (55 %). De plus, les résultats indiquent que les gens étaient peu mobiles avant l’âge de 16 ans. Toutefois, 12 individus ont entrepris des déplacements entre 7 et 16 ans, majoritairement d’un environnement enrichi vers Montréal (N=5) ou de Montréal vers une région appauvrie (N=5). L’âge de recrutement des mousses sur les navires, la traite de la fourrure, la coupe du bois et possiblement aussi l’esclavage pourraient expliquer cette « jeune » migration. Sur le plan alimentaire, les végétaux de type C3, la viande nourrie aux ressources C3 et le poisson faisaient partie du menu montréalais. Les plantes C4 (majoritairement maïs mais aussi sucre de canne [rhum]) étaient consommées en quantité variable. La question de l’influence de la migration sur l’alimentation n’a pu être explorée en profondeur en raison de contraintes liées à la contamination du d18O du carbonate des os. La combinaison des données ostéologiques et isotopiques à la distribution spatiale des sépultures, a permis d’étudier un aspect de l’archéologie funéraire à l’échelle individuelle (identité possible), sans toutefois fournir de résultats probants, à l’échelle du cimetière et de son organisation globale.

Vegetation and environmental dynamics in the southern Black Sea region since 18 kyr BP derived from the marine core 22-GC3

Sediments from the Black Sea, a region historically dominated by forests and steppe landscapes, are a valuable source of detailed information on the changes in regional terrestrial and aquatic environments at decadal to millennial scales. Here we present multi-proxy environmental records (pollen, dinoflagellate cysts, Ca, Ti and oxygen isotope data) from the uppermost 305 cm of the core 22-GC3 (42°13.53′N, 36°29.55′E) collected from a water depth of 838 m in the southern part of the Black Sea in 2007. The records span the last ~ 18 kyr (all ages are given in cal kyr BP). The pollen data reveal the dominance of the Artemisia-steppe in the region, suggesting rather dry/cold environments ~ 18–14.5 kyr BP. Warming/humidity increase during melt-water pulses (~ 16.1–14.5 kyr BP), indicated by δ18O records from the 22-GC3 core sediment and from the Sofular Cave stalagmite, is expressed in more negative δ13C values from the Sofular Cave, usually interpreted as the spreading of C3 plants. The records representing the interstadial complex (~ 14.5–12.9 kyr BP) show an increase in temperature and moisture, indicated by forest development, increased primary productivity and reduced surface run-off, whereas the switch from primary terrigenous to primary authigenic Ca origin occurs ~ 500 yr later. The Younger Dryas cooling is clearly demonstrated by more negative δ13C values from the Sofular Cave and a reduction of pines. The early Holocene (11.7–8.5 kyr BP) interval reveals relatively dry conditions compared to the mostly moist and warm middle Holocene (8.5–5 kyr BP), which is characterized by the establishment of the species-rich warm mixed and temperate deciduous forests in the low elevation belt, temperate deciduous beech-hornbeam forests in the middle and cool conifer forest in upper mountain belt. The border between the early and middle Holocene in the vegetation records coincides with the opening of the Mediterranean corridor at ~ 8.3 kyr BP, as indicated by a marked change in the dinocyst assemblages and in the sediment lithology. Changes in the pollen assemblages indicate a reduction in forest cover after ~ 5 kyr BP, which was likely caused by increased anthropogenic pressure on the regional vegetation.

Stable isotope and modelling evidence for CO2 as a driver of glacial–interglacial vegetation shifts in southern Africa

Atmospheric CO2 concentration is hypothesized to influence vegetation distribution via tree–grass competition, with higher CO2 concentrations favouring trees. The stable carbon isotope (δ13C) signature of vegetation is influenced by the relative importance of C4 plants (including most tropical grasses) and C3 plants (including nearly all trees), and the degree of stomatal closure – a response to aridity – in C3 plants. Compound-specific δ13C analyses of leaf-wax biomarkers in sediment cores of an offshore South Atlantic transect are used here as a record of vegetation changes in subequatorial Africa. These data suggest a large increase in C3 relative to C4 plant dominance after the Last Glacial Maximum. Using a process-based biogeography model that explicitly simulates 13C discrimination, it is shown that precipitation and temperature changes cannot explain the observed shift in δ13C values. The physiological effect of increasing CO2 concentration is decisive, altering the C3/C4 balance and bringing the simulated and observed δ13C values into line. It is concluded that CO2 concentration itself was a key agent of vegetation change in tropical southern Africa during the last glacial–interglacial transition. Two additional inferences follow. First, long-term variations in terrestrial δ13Cvalues are not simply a proxy for regional rainfall, as has sometimes been assumed. Although precipitation and temperature changes have had major effects on vegetation in many regions of the world during the period between the Last Glacial Maximum and recent times, CO2 effects must also be taken into account, especially when reconstructing changes in climate between glacial and interglacial states. Second, rising CO2 concentration today is likely to be influencing tree–grass competition in a similar way, and thus contributing to the "woody thickening" observed in savannas worldwide. This second inference points to the importance of experiments to determine how vegetation composition in savannas is likely to be influenced by the continuing rise of CO2 concentration.

Evidence of a universal scaling relationship for leaf CO2 drawdown along an aridity gradient

The leaf carbon isotope ratio (δ13C) of C3 plants is inversely related to the drawdown of CO2 concentration during photosynthesis, which increases towards drier environments. We aimed to discriminate between the hypothesis of universal scaling, which predicts between-species responses of δ13C to aridity similar to within-species responses, and biotic homoeostasis, which predicts offsets in the δ13C of species occupying adjacent ranges. The Northeast China Transect spans 130–900 mm annual precipitation within a narrow latitude and temperature range. Leaves of 171 species were sampled at 33 sites along the transect (18 at ≥ 5 sites) for dry matter, carbon (C) and nitrogen (N) content, specific leaf area (SLA) and δ13C. The δ13C of species generally followed a common relationship with the climatic moisture index (MI). Offsets between adjacent species were not observed. Trees and forbs diverged slightly at high MI. In C3 plants, δ13C predicted N per unit leaf area (Narea) better than MI. The δ13C of C4 plants was invariant with MI. SLA declined and Narea increased towards low MI in both C3 and C4 plants. The data are consistent with optimal stomatal regulation with respect to atmospheric dryness. They provide evidence for universal scaling of CO2 drawdown with aridity in C3 plants.

Diet and lifestyle in Bronze Age Northwest Spain: the collective burial of Cova do Santo

A multidisciplinary investigation of the collective burial of Cova do Santo is presented as a novel approach to understand daily life during the Bronze Age in Northwest Iberia. The research is focused on three main aspects: i) taphonomy and patterns of disposal, ii) paleopathology and -demography as indicators of health status and lifestyle, and iii) stable isotope analysis to reconstruct paleodiet and to investigate the timing of the introduction of millet to the Iberian Peninsula. Osteological analyses were performed on 64 bones (61 human and 3 animal); additionally, bone collagen was extracted from 15 samples (13 human and 2 animal) and analyzed for its carbon and nitrogen stable isotopes composition. The radiocarbon age of the human remains is consistent with the Middle Bronze Age (c. 1890 to 1600 cal BC). The recovered remains belonged to a minimum number of 14 individuals with an estimated age at death of forty years or younger. This relatively young age is in contrast to a high prevalence of degenerative joint disease in the group. The isotopic results suggest a very homogeneous diet, which was almost exclusively based on C3 plants and terrestrial animal products. Overall, the data suggest that the studied population belonged to a period prior to the introduction of spring or summer-grown crops such as millets. The collective burial from the cave of Cova de Santo, Galicia, currently represents the largest assemblage of prehistoric human remains from Northwest Spain and the relatively good preservation of the bones offers a unique opportunity to investigate daily life in Northern Iberia during the Bronze Age.