岷江干旱河谷三种蔷薇植物繁殖更新研究

繁殖更新是植物生活史的重要阶段，在退化生态系统中，植物繁殖更新能力往往较差，是植被恢复的限制环节，因而也成为恢复研究重点和核心。本研究选择岷江干旱河谷广泛分布的三种蔷薇：多苞蔷薇（R. multibracteata）、黄蔷薇（R. hugonis）和川滇蔷薇（R. soulieana）为研究对象，通过野外调查，在查明其生长、繁殖更新状况的基础上，采用控制和模拟实验，对种子和幼苗阶段进行了深入研究，综合分析更新潜力，并提出相对应的促进更新和植被恢复措施。主要结论如下： 1）三种蔷薇在岷江干旱河谷广泛分布，生长和繁殖状况良好，结实量大。各生长指标：株高、基径和冠幅，繁殖指标：结实数量、重量和单果重量都具有显著的空间差异性。基径对多苞蔷薇结实量影响最大；而冠幅对黄蔷薇结实量影响最大。海拔和纬度是对蔷薇生长和繁殖影响最大的环境因素，随着海拔和纬度的升高，植株生长更高大，结实量增加；坡度和坡向对其生长和繁殖也有一定影响，随着坡度 和坡向增加，蔷薇生长和结实受到抑制。 2）三种蔷薇在岷江干旱河谷更新现状不佳, 但更新潜力大。活力种子比率低，动物取食以及两年生幼苗的大量死亡是蔷薇更新的主要限制因素。多苞蔷薇和黄蔷薇的结实率低，川滇蔷薇较高。三种蔷薇种子产量大，但种子质量较差，更新具有充足的种源。三种蔷薇都能形成持久种子库，种子库中种子总量大，但有效种子少，黄蔷薇被动物啃食的比例很高，多苞蔷薇和川滇蔷薇也有一部分种子受到动物破坏。三种蔷薇幼苗库组成特征表现为，当年生幼苗所占比例很高，年龄较大幼苗所占比例小。 3）三种蔷薇都具有不同程度休眠，未经处理种子的发芽率极低。黄蔷薇休眠程度最深，为深度生理休眠；多苞蔷薇为中度生理休眠；川滇蔷薇为非深度生理休眠。三种蔷薇种子在形态上发育成熟，种皮具有透水性。蔷薇果果肉和瘦果中含有抑制物质，其浸泡液抑制了油菜种子萌发，果肉抑制作用更强，果肉和瘦果浸泡液的抑制程度分别为：川滇蔷薇>黄蔷薇>多苞蔷薇。切割和硫酸腐蚀提高了川滇蔷薇种子的发芽能力，而对多苞蔷薇和黄蔷薇没有影响。完全去除瘦果果皮和种皮提高了多苞蔷薇种子发芽率，但对黄蔷薇没有影响。赤霉素和烟水对蔷薇种子萌发没有促进作用。三种蔷薇打破休眠所需低温层积时间分别为：黄蔷薇>多苞蔷薇>川滇蔷薇。对于多苞蔷薇和川滇蔷薇，层积前对种子进行硫酸腐蚀或暖温层积能缩短低温层积时间，提高发芽率。对于多苞蔷薇，变温层积中暖温层积和低温层积具有一定的负补性，即延长暖温层积可以缩短种子萌发对低温层积的需要。 4）多苞蔷薇种子形态特征和种子休眠与萌发在不同海拔梯度间存在较大差异。种子采集时间、采集季节和干藏影响多苞蔷薇和川滇蔷薇的种子休眠。多苞蔷薇果实大小、种子大小和千粒重、种皮厚度随海拔升高而增加，而种子饱满率和活力随海拔升高而降低，种子休眠程度也随海拔升高而增加。种皮厚度与种子大小、千粒重成正相关关系，硫酸腐蚀后的种子经过不同时间的低温层积后，种子发芽率与种皮厚度、种子大小、千粒重、海拔成正相关关系。2006 年采集川滇蔷薇和多苞蔷薇种子休眠程度较2005 年低。种子休眠随种子年龄增加而减弱。高温和干旱能减轻多苞蔷薇和川滇蔷薇种子休眠。 5）三种蔷薇的生长和生物量积累在干旱胁迫条件下受到抑制，而生物量分配、叶片形态特征和水分利用特征等都发生了变化。三种蔷薇的根、茎、叶各器官生物量以及总生物量等在干旱胁迫下明显减小，叶片脱落数量增加。在干旱胁迫条件下，较多的生物量分配到地下部分，从而这使R/S 明显增加。比叶面积（SLA）和冠层面积比（LAR）对干旱胁迫的反应不敏感，仅有部分物种在干旱胁迫条件下发生了变化，并且其变化特点在不同年龄幼苗之间有一定差异。干旱胁迫对WUE 的影响在不同物种间存在差异。多苞蔷薇和黄蔷薇的WUE 随着干旱胁迫的增加而增大， 而川滇蔷薇的WUE 则随干旱胁迫增加而减小。在干旱胁迫条件下，多苞蔷薇和黄蔷薇叶片脱落量和生物量减小幅度较川滇蔷薇大，表明其抗旱能力较强。在干旱胁迫条件，三种蔷薇两年生幼苗的生物量减小幅度较当年生幼苗小，表明两年生幼苗的抗旱能力更强。 6）两种植被恢复措施中，幼苗移栽比播种具有更好的植被恢复效果。播种后，蔷薇种子的发芽率较高，但出苗率都很低，即使出苗，幼苗也几乎在一月内全部死亡。 三种微生境条件下（灌木、半灌木和裸地），种子出苗和幼苗成活没有差异。移栽幼苗总体死亡率都比较低，小于20％。特别是两年生幼苗死亡率更低，小于2％。移栽后的幼苗生长状况良好，在整个生长季中，各生长指标不断增加。生境对幼苗的存活率没有显著影响，但对于幼苗的生长和生物量积累有一定影响，裸地更有利于幼苗生长和生物量积累。与当年生幼苗相比，两年生幼苗具有更高的成活率。总之，三种蔷薇在干旱河谷分布广泛、生长繁殖状况良好，结实量大，具有丰富种源，繁殖更新潜力大，但繁殖更新状况不佳；种子散布后动物对种子的取食、种子的深度休眠过程、种子出苗以及当年生幼苗的存活和定居是更新的主要限制环节。水分是影响结实、种子休眠解除和萌发，幼苗存活和定居的最主要的限制因素。在植被恢复中，应在种子成熟季节大量采集种子，在室内打破休眠后进行人工播种，培育两年生幼苗，通过幼苗移栽方式进行植被恢复。川滇蔷薇应栽种在相对湿润的过渡区，而多苞蔷薇和黄蔷薇可以应用于核心区植被恢复。 Regeneration is an important phase in plant life cycle. It has been a key component of ecological restoration in degradation ecosystem in which plants commonly has poor regeneration. In this paper, we investigated the natural growth, propagation and regeneration status of native three rose species, Rosa multibracteata, R. hugonis and R. soulieana, and analysis the limitation in seed germination and seedling establishment stages. Advice on facilitating the use of these plants in restoration based on the results has been proposed. The results were as follows: 1) Three rose plants widely distributed in the dry valley of the Minjiang River, and made a good performance in growth and propagation. There were significant spatial differences in each growth parameter, such as ramet height, basal diameter, crown diameter and propagation parameters including hip number of a clump, hip mass of a clump and a hip mass. Basa diameter was the most important growth parameter influencing fruit number for R. multibracteata and crown diameter was for R. hugoni. Altitude and latitude had the greatest effect on the growth and propagation of rose plants among environmental conditions. Each parameter of growth and propagation increased with the increase of altitude and latitude. In addition, the increase of slope and aspect limited the growth and propagation. 2) Three rose plants had poor natural regeneration, but great regeneration potential. Low seed viability, predation and higher mortality of current year old seedlings were the limitation in regeneration. R. multibracteata and R. hugonis had higher fruiting rates than R. souliean. All three plants produced a great number of seeds, while their viability was poor. Three rose plants had persistent seed banks, with high total seed number but very low viable seed density. Predation was most severe in R. hugonis, and it also existed to some degree in R. multibracteata and R. soulieana. The seedling age-structure was characteristic of current-year seedlings predominating and few older seedlings were observed. 3) Three rose seeds were dormant and untreated seeds germinated with very low germination percentages. The rose seeds had morphological mature embryos, and achenes were permeable. Some inhabit substances existed in hips and achenes for the extracts of hips and achenes inhibited germination of Brassica campestris. The inhibition effect of the extracts of three rose hip and achenes was R. soulieana>R. hugonis>R. multibracteata. Mechanical and H2SO4 scarification increased R. soulieana germination but had no effect on germination of R. hugonis and R. multibracteata seeds. Full removal of pericarp and testa improved the germination of R. multibracteata but did not affect R. hugonis germination. GA3 and smoke water had no positive effect on rose seed germination. The periods of cold stratification required to released seed dormancy was R. hugonis > R. soulieana >R. multibracteata. H2SO4 scarification and warm stratification shortened cold stratification to release dormancy for R. soulieana and R. multibracteata. Warm stratification had complementary effect for cold stratification, i.e. the longer warm stratification seeds received, the shorter cold stratification were required to obtain the same germination percentage. Three rose seeds had different kinds of dormancy; R. hugonis has deep physiological dormancy, R. multibracteata with intermediate physiological dormancy and R. souliean non-deep physiological dormancy. 4）The seeds traits and dormancy of R. multibracteata showed significant difference across altitudes. Year and season of seed collection had significant effect on seed dormancy for both R. souliean and R. multibracteata. Hip size, seed size, seed weight, seed coat thickness and seed dormancy level increased with the increase of the altitude. There were positive relations between seed coat thickness with seed size and seed weight. Germination percentage of seeds treated with H2SO4 scarification following different periods of cold stratification showed positive relation with seed coat thickness, seed size, seed weight and altitude. Seeds of R. souliean and R. multibracteata collected in 2006 had low dormancy level than those collected in 2005. Seed dormancy decreased with increasing seeds age. High temperature and drought were associated with low dormancy level. 5) Seedling growth, the total dry mass and their components of seedlings were reduced, while leaf senescence accelerated under drought stress. More biomass allocation to root system resulted in higher R/S ratio under drought. Water-use efficiency (WUE) of R. multibracteata and R. hugonis increased, while it declined for R. soulieana under drought stress. R. soulieana seedlings had poor drought-resistance capacity it had more senescent leaves, and its reduction of biomass was stronger than two other rose plants under drought. The reduction degree of one year old seedlings under drought stress was slighter than that of current year seedlings. Therefore, one year old seedling was more drought-resistent compared to current year seedlings. 6）Planting seedlings may have better effect in comparison with direct seeding. Most seeds germinated after seeding, but seedling emergence was very low. More than 80 % seedlings from direct seeding died within a months after emergence. Seedling emergence and survival rate did not show difference among microhabitats. Mortality rates of seedlings artificially planted in microhabitats were general lower than 20 %, and the mortality rate of one year old seedlings was lower than 2 %. Each grow parameter including plant height, leaf number and branch number continually increased after planting. Microhabitat type had effect on the growth parameter and biomass production, but it did not influence the seedling survival. Bare land tended to facilitate seedling growth. One year old seedlings had higher survival rate than current year seedlings. In conculsion, the three rose had wide distribution in the dry valley of the Minjiang River. They produced many seeds and had tolerance to drought stress to some degree. But they had poor regeneration in habitats may be caused by predation, seed dormancy，and high mortality in current year seedlings. We recommend that rose plants should be utilized in restoration by planting two-year old seedlings in spring. A large quantity of seeds should be collected artificially in autumn, release seed dormancy in room, and then cultivate two-year old seedlings by seeding in particular container. R. soulieana seedling probably be planted in transition area, and R. multibracteata and R. hugonis can be used in core area of the dry valley of the Minjiang River.

九寨沟核心景区湖泊硅藻的组成及其密度与环境因子的关系

九寨沟自然保护区地处四川盆地北部岷山山脉南段，属川西高原气候中的暖温带。保护区内分布着118 个串珠状排列的高山湖泊，核心景区共有湖泊20 个（组），集中在海拔2000 米到3000 米之间。水资源是九寨沟的灵魂，保护水生生态系统是九寨沟生态旅游可持续发展的立足之本。本研究以九寨沟湿地生物多样性保护项目为依托，于2007 年5 月～2008 年4 月对九寨沟核心景区湖泊水体硅藻植物进行调查，分析了其密度与环境因子的相互关系，旨在为九寨沟湖泊生态系统保护提供科学依据和基础资料。主要有以下几点结论： （1）九寨沟硅藻植物丰富，调查统计为2 纲6 目10 科35 属159 种（包括变种和变型），水体硅藻中真性浮游种类较少，着生种类多，羽纹纲的脆杆藻属、舟形藻属、桥弯藻属、异极藻属、曲壳藻属的种类最多，常为优势属组成湖泊硅藻群落，只在长海是小环藻属占绝对优势。 （2）九寨沟水体中硅藻细胞密度不大，多在4×104～30×104 个/L，长海最高，五花海最低。在年变化上，不同的湖泊水体中硅藻的生长高峰季节不尽相同，在夏季、秋末和冬末春初均有可能出现高密度。在空间上，整体看来密度与海拔有一定的正相关，但由于水温不是单一主导因素，密度受多种环境因子影响，在海拔梯度上呈现一定的波浪状变化。 （3）影响九寨沟湖泊水体硅藻细胞密度较重要的环境因子是海拔、水温和pH。硅藻细胞密度年变化幅度大的湖泊水体，年平均细胞密度大。因此，环境因素不太稳定的湖泊水体，更可能出现高的硅藻细胞密度。11 月份响应于水量剧烈的减少，细齿菱形藻大量出现在五彩池成为优势种群，造成水体硅藻细胞密度的大幅增加。 （4）九寨沟湖泊水体水质几乎达到国家地表水I 类标准，但是，硅藻商指数以及代表多样性的Cairns 指数计算的结果不能反映水质的情况，这是由于水质状况不是决定九寨沟湖泊水体中硅藻生存的种类和数量的主要因素。因此在用各种指数分析水质的时候，都应该先判断计算该指数的参数是否主要由水质状况决定，然后再综合理化性质等其他指标分析和比较。 （5）人类的活动对九寨沟湖泊水生态系统有一定的影响。一方面，在低海拔景点人类活动的干扰比海拔高处相对要大，干扰引起低海拔景点湖泊硅藻Cairns指数较高；另一方面，旺季的影响比淡季大，水体中出现了典型富营养种类。因此，应加强低海拔景点和旺季的管理，并限制淡季的人类活动以保证九寨沟生态系统的自我恢复。 Jiuzhaigou nature reserve is located in north Sichuan, at the south section of Minshan Mountain, having a warm temperate climate which belongs to West Sichuan plateau climate. It includes 118 mountain lakes, and 20 of them between 2000 and 3000 altitude degree constitute the core area of Jiuzhaigou. The water is Jiuzhaigou's soul, so it is the basis of sustainable tourism development of Jiuzhaigou to protect the water ecological environment. Research depends on Jiuzhaigou wetland biodiversity protection program in this paper, invested the diatom plants in 17 lakes at the core area from May 2007 to April 2008. The composition and the correlation of diatom density and the environment were studied, which could provide scientific and basic information for the protection of the ecological environment of Jiuzhaigou. The main conclusions were: 1 The diatom species were in abundance, investigation showed that there were 159 species belong to 10 families and 35 genera. Most diatoms were benthos, euplankton were less. Genera Fragilaria, Navicula, Cymbella, Gomphonema and Achnanthes had the most species, which always were the dominant genera. Only in The Long Lake genus Cyclotella possessed a predominant status. 2 The diatom cell density was not very high, always between 4×104 cell/L and 30×104 cell/L, highest in The Long Lake and lowest in The Colorful Lake. In one year's time, diatom in different lakes had different growth type. The maximum could be in summer, late autumn and late winter early spring. The cell density showed a positive correlation to altitude in the whole, but fluctuated in details along altitude because of comprehensive influence by many environmental factors. 3 Altitude, water temperature and pH were the more important factors to determine the cell density. There was a positive correlation between the range of cell density in one year and the average cell density. So high annual cell average density probably will appear in lakes which has an unstable environmental condition. The boom of Denticula elegans in The Jade-Colored Lake in November was corresponding to the large decrease in water quantity. 4 The water quality of Jiuzhaigou was near the National Surface Water Standard rank I, but the Diatom Quotient and the Cairns Index didn't reflect the water quality accordingly. It was because the water quality wasn't the most important factor here which determined the species number and species quantity, so the index based on these items would not reflect it. Therefore, it is recommended that, when we use indices to evaluate the water quality, we should know first whether the water quality is the main determinative factor inflects the items used to compute the indices, and then combined with other chemical indices to analyze and do comparison. 5 The tourists' activities had influenced the water ecosystem of Jiuzhaigou. On the one hand, disturbance was relatively high at the lower altitude, it resulted in higher Cairns index; on the other hand, influence in high season was more severe than in low season, some typical species indicating eutrophication were observed in the water. It is suggested that the management at lower altitude and in high season must be strengthen, and less people activity in low season is necessary to fulfill the recovery of the water ecosystem.

川西亚高山两种针叶树幼苗对增温和施氮的响应

人类活动引起全球大气中温室气体（CO2、CH4、NOx）浓度不断增加，致使地球表面温度在过去的100 年中已经增长了0.74 ± 0.18℃，预计到本世纪末将会增加1.1-6.4℃。此外，氮沉降也是当今社会的重要环境问题，随着经济发展的全球化, 高氮沉降也呈现出全球化趋势。全球气候变暖和氮沉降给陆地生态系统的地上、地下生物学和生物地球化学过程所带来巨大影响越来越引起人们的关注。 本文以川西亚高山针叶林的两个重要树种云杉和油松幼苗为研究对象，采用红外辐射增温（空气增温2.1℃，土壤增温2.6℃）和根部施氮（施氮量25 g N m-2yr-1）的方法，从生长形态、光合作用、抗氧化能力和矿质营养等方面研究这两种幼苗对气候变暖和氮沉降的响应。该实验为室外控制实验，包括四个处理：(1)不增温+不施氮（UU）；(2) 不增温+施氮（UF）；(3) 增温+不施氮（WU）；(4) 增温+施氮（WF）。本研究旨在从生理生化、物质代谢 、生长及形态等不同水平上研究模拟增温和施氮对两种树苗的联合效应，提高我们对全球变化下亚高山针叶林早期更新过程的理解，同时也为森林管理提供科学依据。具体研究结果如下： 单独增温处理显著提高了云杉和油松幼苗的地茎、叶重、茎重、根重以及总生物量；单独施氮处理也增加了两种幼苗的株高和总生物量。而增温和施氮联合作用对两种幼苗生长的影响并不相同，联合作用对云杉幼苗生长指标的正效应显著低于单独施氮处理，但是联合作用比单独增温或施氮更大程度的促进了油松幼苗生物量的积累。 单独增温和施氮都有利于提高云杉和油松叶片中叶绿素含量、净光合速率(A)、最大净光合速率(Amax)、表观量子效率（Φ）、最大光能转化效率（Fv/Fm）和量子产量（Y）。与对两种幼苗生长指标的影响相似，加氮和增温共同作用下油松幼苗的以上光合指标比在单独增温或施氮处理下有更大程度的提高；而联合作用下云杉幼苗叶绿素含量、净光合速率、最大净光合速率、表观量子效率、最大光能转化效率以及量子产量比单独施氮处理明显地降低。 增温和施氮都显著地降低了云杉和油松幼苗针叶组织中活性氧和丙二醛的积累。交互作用降低了云杉幼苗叶片的抗氧化酶活性、脯氨酸和ASA 的含量，却显著提高了油松幼苗SOD、POD、APX 等抗氧化酶的活性，并且对油松幼苗脯氨酸和ASA 积累的促进作用比单一因子更加明显。因此，增温和施氮共同作用下油松幼苗叶片中O2-产生速率、H2O2 及MDA 含量明显降低，而云杉叶片中只有O2-产生速率出现降低趋势。 增温和施氮都降低了云杉体内的P、Ca、Mg 元素的含量，增加了Cu、Zn、Mn 在各器官内的积累。对油松幼苗而言，增温和加氮单独作用也显著降低了Ca 含量增加了Cu、Zn、Mn 的积累，但是不同于云杉幼苗的是P、Mg 也显著增加。增温和施氮联合作用对云杉幼苗体内元素的影响与单一施氮处理或增温处理相似，不同的是比单一因子作用更为明显降低了P、Ca、Mg 含量，增加了植株中N、Cu、Zn、Mn 的含量，但是油松矿质元素含量在联合作用下并没有产生类似于云杉幼苗的双因子叠加效应。 总之，尽管单独增温或者施氮都有利于云杉和油松幼苗生长指标、光合能力以及抗氧化能力的提高。但是，增温和施氮对云杉幼苗生长生理的促进效应非但没有在交互作用下有更大的提高，反而低于单独氮处理。与此不同的是，增温和施氮联合作用比单因子作用更有利于油松幼苗生长及生理指标的提高。 With the continued increase in atmospheric concentrations of greenhouse gases (CO2、CH4、NOx), the mean global surface temperature has increased by about 0.74 ± 0.18℃ over the past century and is predicted to rise by as much as 6.4℃ during this century. Besides global warming, nitrogen deposition is another serious environmental problem caused by human activities, and high nitrogen load has become globalization as a result of global economy development. Global climate warming and nitrogen deposition have induced dramatic alternations in above - and below- ground biology and biogeochemistry process in terrestrial ecosystems, and more and more attention has been invited to those problems. This experiment mainly studies two important species Picea asperata and Pinus tabulaeformis in subalpine coniferous forest of western Sichuan, China. Infared heaters are induced to increase both air and soil temperature by 2.1℃ and 2.6 ℃, respectively. Ammonium nitrate solution (for a total equivalent to 25 g N m-2 year-1) is added to soil surface. There are four treatments in this study: (1) unwarmed unfertilized (UU); (2) unwarmed fertilized (UF); (3) warmed unfertilized (WU); (4) warmed fertilized (WF). This study is conducted to determine the influences of experimental warming and nitrogen fertilization on physiolchemistry, nutrition metabolism, growth and morphology in the two coniferous species seedlings. The current study is favorable for increasing our understanding on the early phase of regeneration behavior in subalpine coniferous forest, and it also provide scientific direction for forest management under future global changes. The results are as follows: Artificial warming alone significantly increased basal diameter, leaf mass, stem mass, root mass and total biomass for Picea asperata and Pinus tabulaeformis seedlings, and single nitrogen fertilization are also favorable for growth of the two species and stimulate plant hight and total biomass. The two species seedlings respond differently to the combination of elevated temperature and nitrogen addition. Warming combined with nitrogen fertilization weakens the positive effects of nitrogen addition for growth of Picea asperata seedlings. However, the combination of elevated temperature and nitrogen fertilization further increase biomass accumulation of Pinus tabulaeformis seedlings. Both elevated temperature alone and nitrogen fertilization alone can increase photosynthetic pigments contents, net photosynthetic rate (A), maximum net photosynthetic rate (Amax), apparent quantity yield (Φ), maximum photochemical efficiency of photosystem II (Fv/Fm) and effective quantum yield (Y). Similarly with growth parameters, the combination of warming and nitrogen addition induced more increment of these above photosynthetic parameters for Pinus tabulaeformis seedlings. However, these photosynthetic parameters of Picea asperata seedlings under the combination of warming and nitrogen addition are lower than those under nitrogen fertilization alone. The levels of active oxygen species (AOS) and malodiadehyde (MDA) in needles of the two coniferous species seedling are obviously decreased by experimental warming or additional nitrogen. Warming combined with nitrogen fertilizer reduces the activities of SOD, CAT and APX, and the contents of proline and ASA of Picea asperata seedlings, but the combination significantly increases activities of these antioxidant enzymes in needlels of Pinus tabulaeformis seedlings and further improves the accumulation of proline and ASA compared to either artificial warming or nitrogen addition. Therefore, the rate of O2 - production, the contents of H2O2 and MDA in needles of Pinus tabulaeformis seedlings are remarkably reduced by the combination of warming and nitrogen addition, but the combination only significantly decreased the rate of O2 - production of Picea asperata seedlings. Elevated temperature or nitrogen fertilization decrease the contents of P, Ca, Mg but increase Cu, Zn, Mn contents for Picea asperata seedlings. For Pinus tabulaeformis seedlings, elevated temperature alone and nitrogen fertilization alone decreased Ca, but increased P, Mg, Cu, Zn, Mn contents. The effects of the combination of warming and nitrogen addition on these element contents in needles of Picea asperata seedlings are added or multiplied the effects of warming and nitrogen addition alone, resulting in less contens of P, Ca, Mg and more contents of Cu, Zn, Mn than either elevated temperature or nitrogen fertilization. Howere, these adding or multipluing single-factor effects on contents of these elements are not observed in the case of Pinus tabulaeformis seedlings. In conclusion, growth parameters, photosynthetic capacities and antioxidant abilities of Picea tasperata and Pinus abulaeformis seedlings are improved by experimental warming or nitrogen fertilization. Interestingly, the positive effects of warming and nitrogen addition on growth and physiological performances are not multiplied by the combination of elevated temperature and nitrogen fertilization, even dempened for Picea asperata seedlings. However, for Pinus tabulaeformis seedlings, growth and physiological performances are further improved by the combination.

青稞B组醇溶蛋白基因与上游调控区的克隆及基因表达

高等植物种子胚乳贮藏蛋白是种子发芽时的主要氮源，也是人类和动物食用植物蛋白的主要来源。大麦种子胚乳贮藏蛋白主要是醇溶蛋白（hordeins），占大麦胚乳总蛋白的50–60%。根据大麦醇溶蛋白的大小和组成特点，大麦醇溶蛋白被划分为三种类型：富硫蛋白亚类(B，γ-hordeins)、贫硫蛋白亚类（C-hordeins)以及高分子量蛋白亚类(D-hordeins)。B组和C组醇溶蛋白是大麦胚乳的两类主要贮藏蛋白，它们分别占大麦总醇溶蛋白成分的70–80%和10–12%。遗传分析表明，大麦B、C、D和γ-组醇溶蛋白分别是由位于大麦第五染色体1H（5）上的Hor2、Hor1、Hor3和Hor5位点编码。Hor2位点编码大量分子量相同但组成不同的B组醇溶蛋白（B-hordein）。B-hordein的种类、数量和分布是影响大麦酿造、食用及饲养品质的重要因素之一。为深入了解B-hordein基因家族的结构和染色体组织，探明Hor2位点基因表达的发育调控机制，最终达到改良禾谷类作物籽粒品质的目的，本研究以青藏高原青稞为材料，采用同源克隆法，分别克隆B-hordein基因和启动子，通过原核生物表达验证B-hordein基因功能，并利用实时定量PCR探索B-hordein基因表达时空关系，取得如下研究结果： 1. 以具有特殊B组醇溶蛋白亚基组成的9份青藏高原青稞为材料，根据GenBank中三个B-hordein基因序列（GenBank No. X03103, X53690和X53691）设计一对引物，通过PCR扩增，获得23个B-hordein基因克隆并对其进行了序列分析。核苷酸序列分析表明，所有克隆均包含完整的开放阅读框。有11个克隆都存在一个框内终止密码子，推测这11个克隆可能是假基因。推测的氨基酸序列分析表明，所有大麦B-hordein具有相似的蛋白质基本结构，均包括一个高度保守的信号肽、中间重复区以及C-端结构域。不同大麦种重复区内重复基元的数目有较大差异。青稞材料Z07–2和Z26的B-hordeins仅具有12个重复基元结构，更接近于野生大麦。这些重复基元数目的差异导致了重复区序列长度和结构的变异。这种现象极可能是由于醇溶谷蛋白基因在进化过程中染色体的不平衡交换或复制滑动所造成的。对所克隆基因和禾本科代表性醇溶谷蛋白基因进行聚类分析，结果表明所有来自栽培大麦的B-hordeins聚类成一个亚家族，来自野生大麦的B-hordeins以及普通小麦的LMW-GS聚类成另外一个亚家族，表明这两个亚家族的成员存在显著差异。此外，我们发现B-hordein基因推测的C-末端序列具有一些有规律的特征：即具有相同C-末端序列的B-hordein基因在系统发生树中聚类为同一个亚组（除BXQ053，BZ09-1，BZ26-5分别单独聚为一类外）。这个特征将有助于我们对所有B组醇溶蛋白基因家族成员进行分类，避免了在SDS-PAGE电泳图谱上仅依靠大小分类的局限性。 2. 根据上述克隆的青稞B-hordein基因的5’端序列设计三条基因特异的反向引物，以青稞Z09和Z26的基因组DNA为模板，采用SON-PCR和TAIL-PCR技术分离克隆出8个B-hordein基因的上游调控序列（命名为Z09P和Z26P）。序列分析表明，推测的TATA box位于–80 bp，CAAT–like box位于–140 bp处。此外，Z09P和Z26P中有六个序列在–300 bp处均存在一个由高度保守的EM基序和类GCN4基序构成的胚乳盒(Endosperm Box，EB)，在约–560 bp处存在一个胚乳盒类似结构。而Z09P-2和Z26P-3不存在保守的胚乳盒或其类似结构，预示着这两个启动子所调控的基因表达可能受不同类型反式作用因子的调节，推测该启动子对基因的表达调控具有多样性。 3. 将B-hordein基因的开放阅读框定向克隆到表达载体pET-30a中，将其导入大肠杆菌表达菌株BL21中进行外源基因的诱导表达以验证所克隆基因的功能。结果表明仅含重组子pET-BZ07-2和pET-BZ26-5的BL21细菌有目的表达蛋白产生。在诱导3 h时的蛋白表达量最高；3 mM IPTG诱导的蛋白表达量要高于1 mM IPTG诱导的表达量。这为分离纯化B-hordein蛋白以及进一步研究其对大麦籽粒品质的影响奠定基础。 4. 根据从青稞Z09和Z26中分离克隆的B-hordein基因序列设计一对基因特异的引物，同时，选择大麦α-微管蛋白基因（GenBank no. U40042）为看家基因并设计特异引物，利用实时荧光定量PCR检测了青稞籽粒4个胚乳发育时间段的B-hordein基因表达，荧光定量结果显示：两份材料中B-hordein基因的表达量均随发育过程的进行而逐渐升高。Z09中B-hordein基因在开花后7天开始转录，而Z26开花4天后就有低水平B-hordein的表达，这表明Z26中B-hordein基因可能比Z09表达的较早或者Z09中B-hordein基因表达水平较低以致于不能被检测到。此外，在4个不同的胚乳发育时期中，Z26中B-hordein基因的表达量均高于Z09材料。在开花12天到18天的过程中，Z09和Z26中B-hordein基因的表达水平有一个急剧性的升高。这说明在不同胚乳发育时期，Hor2位点的B-hordein等位基因变异体存在mRNA的差异表达。 Seed endosperm storage proteins in higher plants are the main resources of nitrogen for germinating and plant proteins for human and animals. Barley prolamins (also called hordeins) are the major storage proteins in the endosperm and account for 50–60% of total proteins. Hordeins are classically divided into three groups: sulphur-rich (B, γ-hordeins), sulphur-poor (C-hordeins) and high molecular weight (HMW, D-hordeins) hordeins based on the size and composition. B-hordeins and C-hordeins are two major groups and each respectively account for about 70-80% and 10-12% of the total hordein fraction in barley endosperm. Genetic analysis showed that B-, C-, C-, γ-hordeins are encoded by Hor2, Hor1, Hor3 and Hor5 locus on the chromosome 1H (5). Hor2 locus is rich in alleles that encode numerous heterogeneous B-hordein polypeptides. It is reported that B-hordein species, quantity and distribution are significant factors affecting malting, food and feed quality of barley. To understand comprehensively the structure and organization of B-hordein gene family in hull-less barley and explore the developmental control mechanisms of Hor2 locus gene expression and eventually to better exploitation in crop grain quality improvement, we isolated and cloned B-hordein genes and promotors of hull-less barley from Qinghai-Tibet Plateau by PCR, and testified their expression founction in bacteria expression system and explore their spatial and temporal expression pattern by quantitative real time PCR. Our results are as followed, 1. Twenty-three copies of B-hordein gene were cloned from nine hull-less barley cultivars of Qinghai-Tibet Plateau with special B-hordein subunits and molecularly characterized by PCR, based on three B-hordein genes published previously (GenBank No. X03103, X53690 and X53691). DNA sequences analyses confirmed that the six clones all contained a full-length coding region of the barley B-hordein genes. Eleven clones all contain an in-frame stop codon and they are probably pseudogenes. The analysis of deduced amino acid sequences of the genes shows that they have similar structures including signal peptide domain, central repetitive domain, and C-terminal domain. The number of the repeats was largerly variable and resulted in polypeptides in different sizes or structures among the genes. Twelve such repeated motifs were found in Z07–2 and Z26, and they are close to those of the wild barleys, and it is most probably caused by unequal crossing-over and/or slippage during replication as suggested for the evolution of other prolamins. The relatedness of prolamin genes of barley and wheat was assessed in the phylogenetic tree based on their polypeptides comparison. Our phylogenetic analysis suggested that the predicted B-hordeins of cultivated barley formed a subfamily, while the B-hordeins of wild barleys and the two most similar sequences of LMW-GS of T. aestivum formed another subfamily. This result indicated that the members of the two subfamilys have a distinctive difference. In addition, we found the B-hordeins with identical C-terminal end sequences were clustered into a same subgroup (except BXQ053，BZ09-1 and BZ26-5 as a sole group, respectively), so we believe that B-hordein gene subfamilies possibly can be classified on the basis of the conserved C-terminal end sequences of predicted polypeptide and without the limit of SDS-PAGE protein banding patterns. 2. The specific primers were designed according to the published sequences of barley B-hordein genes from Z09 and Z26. Using total DNA isolated from them as the templates, eight clones (designated Z09Pand Z26P) of upstream sequences of the known B-hordein genes was obtained by TAIL-PCR and SON-PCR. Sequences analysis shows that the putative TATA box was present at position –80 bp and CAAT-like box at position –140 bp. Besides, a putative Endosperm Box including an Endosperm Motif (EM) and a GCN4-Like Motif was found at position –300 bp in six clones, and another Endosperm-like box was found at positon –560 bp. While the Endosperm Box or Endosperm-like box was not found in Z09P-2 and Z26P-3. This may indicate that gene expression drived by the two promtors was probably controlled by different trans-acting factors and the genetic control mechanism of corresponding gene expression may be diverse. 3. The B-hordein genic region coding for the mature peptide was cloned into expression vector pET-30a and transformed into bacterial strain BL21 for identifying gene expression fountion. Protein SDS–PAGE analysis showed that only the transformed lysate with the pET-BZ07-2 and pET-BZ26-5 constructs produced proteins related to B-group hordeins of barley, and the mounts of proteins induced by 3 mM IPTG and 3 h were higher than other conditions. This established a base for isolating and putifying B-hordein and further exploring their effects on barley grain quality. 4. The gene-specific primers of B-hordein genes from Z09 and Z26 were used for the quantification of B-hordein gene expression. The α-tubulin gene from Hordeum vulgare subsp. vulgare (GenBank accession number U40042) was used as a control gene. The result shows the transcription of the B-hordein genes in Z09 was found 7 days after flowering, while the transcription of the B-hordein genes in Z26 was found 4 days after flowering, but at a very low level, and it suggested that the B-hordein genes in Z26 probably expressed earlier than those in Z09, or the B-hordein genes in Z09 expressed at so a lower level than Z26 that it can not detected. In addition, B-hordein genes in Z26 accession showed higher expression levels than those in Z09 in four developing stages. Furthermore, a progressive increase in the expression levels of the B-hordein genes between 12 and 18 days after anthesis was observed in both Z09 and Z26. It implies that the B-hordein allelic variants encoded by Hor2 locus exist the differential expression in mRNA levels of during barley endosperm development.

不同小麦品质类型与生态环境关系的研究

小麦（Triticum aestivum L.）是世界上种植面积最大，总产量最高，食物加工种类最丰富的粮食作物，占世界人口35 ％－40 ％的人们以此为主要食物。因此小麦产量的高低和品质的优劣直接影响人们对食物需求的安全和满意程度，也影响着人类的营养平衡以及面粉和食品加工业的发展。随着生活水平的提高，人们对于小麦的品质越来越重视。培育优质专用小麦新品种，制定优质专用小麦品种品质生态区划，从而在不同程度上实现小麦的区域化种植和产业化经营具有重要的意义。 影响小麦品质的因素主要是遗传因素和环境因素，其中环境因素又包括各种自然生态因素和人为因素。研究表明，小麦品质的环境间的差异大于品种间的差异，气候条件是影响小麦品质的最重要的因子，小麦品质的地域间的差异反映出了小麦的品质区域分布规律。为了满足市场对不同品质小麦的需求，对小麦进行区域化研究具有重要的理论和现实意义。本研究结合四川的地理、气候特点，研究不同品质类型与生态环境的关系，为在复杂的生态环境内进行品质区划提供依据。 本研究首先根据四川省小麦种植区域的生态特点，在四川省多个典型生态区：川南丘陵的荣县、川西南高原的西昌、川西平原的双流布点种植，采用的小麦试验材料为不同品质类型：中筋小麦川育12、川育14、川育16由本所提供；弱筋小麦川麦32和强筋小麦川麦36由四川省农科院作物所提供。通过研究品质性状与品种及各个生态因子包括地点、土壤土质差异等的关系，明确不同生态环境中适宜种植的小麦品种类型，强筋小麦、中筋小麦更适合于在荣县、双流地区种植，弱筋小麦更适合于在西昌地区种植，为品种品质区划奠定基础。 其次，选择了本课题组育成的稳定中间品系，对其品质性状SDS沉降值进行了多年测定。分析了品质性状SDS沉降值与多种气候因子的相关性，结果表明SDS沉降值与日均温、日照时数成正相关，与降水量成负相关，为品质育种提供了理论依据。 此外，以中筋小麦新品种小麦川育14为材料，应用三元二次正交旋转回归模型设计试验，研究主要栽培因子播期、密度和施肥量对产量的影响，并建立函数模型。经计算机模拟寻优，筛选出了高产高效栽培组合措施，并确定了置信域。结合四川省不同的地理情况，在平原和丘陵地区分别进行实验，并各自建立了高产高效栽培组合措施，为川育14品种的推广提供了理论指导。 Wheat is one of the most important crops in the world. About 35%-40% people all over the world, take the wheat as their most important food. So the quality, as well as the quantity of the wheat makes a direct effect on people’s demands of food and their satisfaction. It also effects on human’s healthy, and the development of the Food processing industry. With the development of the living standard, people pay more attention to the quality of wheat. So, we set a special ecology zoning for wheat. It is significant to carry out planting the wheat in special zoning in varying degrees. The main factors affecting wheat quality are heredity and environment including many ecological factors and the factors in cultivation. As to the quality，the difference between ecology and cultivation is more important than the difference between special wheat. In so many factors, climate is the most important one. From the difference in quality between different zones，we can conclude the rule of distribution abort quality of wheat. Finding out the intersection of numerous wheat not only can meet the demand of food production，but also has important signification in theory and realism。In our research, according to the complex geography in Sichuan province, we study the relationship between numerous kinds of quality characters in wheat and the ecology. So, we can set a foundation for more research. In this research, firstly, we plant wheat in some typical ecological regions of SICHUAN province: RONGXIAN（south of SC）、XICHANG（south of SC）, SHUANGLIU(west of SC). The materials of the experiments: ChuanYu12, ChuanYu14, ChuanYu16(from our institute), Chuanmai32, Chuanmai36 (from the Chinese academy of agriculture sciences of Sichuan. Through the research on the relationship between the quality of wheat and those ecology factors, we can make a definition that which area is perfect matched with which kind of wheat. And it can satisfy the demand of people. Secondly, select many sorts of wheat from our research group. All of them are selected and bred more than 3 years(2003-2005). And we make every-year determination as well. We’ve gotten SDS value from those 9, and various data on factors of climate. We also got to know the relation ship between those numbers. Thirdly, use Chuanyu14 as material, the mathematical model of the relation between the production of wheat and main agricultural measures such as date, density and fertilizer. The model was established by association of three elements two return, rotate and regression. We set a suitable model and get a suitable method which can make high harvest. Based on various kinds of geographical regions in Sichuan province, we set different models which can be used in plain and hill. So, we can plant Chuan Yu 14 in Sichuan province under the result in research.

小麦胚乳蛋白组分和Glu-B3位点基因核心编码区差异与小麦品质关系的研究

小麦加工品质改良已成为我国小麦育种的主要目标之一。特别是我国加入WTO以后，对小麦产品的质量提出了更高的要求，小麦品质改良的任务将更加艰巨和重要，小麦胚乳蛋白是影响小麦加工品质性状的重要因素。因此，深入了解小麦胚乳蛋白对加工品质性状的影响及其分子基础，为品质改良提供理论依据和科学指导，对加速我国小麦品质育种和优质小麦生产具有重要意义。本研究选用在麦谷蛋白5个基因位点(Glu-A1、Glu-B1、Glu-D1、Glu-B3和Glu-D3)上均含不同等位基因的小麦品种99G45和京771及Pm97034和京771杂交F9代共164个麦谷蛋白纯合系，及228个中国推广普通小麦品种和高代育成品系为试材，研究了麦谷蛋白Glu-1和Glu-3位点基因等位变异对籽粒蛋白、湿面筋含量、Zeleny沉降值和SDS沉降值间的关系；本研究还利用小麦A、B和D基因组中低分子量麦谷蛋白亚基（LMW-GS）基因特异引物，通过PCR方法克隆了1个Glu-A3位点和3个Glu-B3位点LMW-GS基因片段，在此基础上分析了不同等位基因对品质造成差异的分子基础；另外，本研究对中国近年推广的部分品种和育成的高代品系资源的多样性进行了分析。现将主要研究结果简述如下： 1. 对来自三个麦区的148份材料的醇溶蛋白组成进行了分析，结果表明，各麦区醇溶蛋白模式具有较大差异。在ω区,A7、B、E、F、G、J、P、Q、S和U仅存在于西南秋播麦区；A3、M、N、R、W和X仅存在于黄淮特种麦区；K仅存在于北方冬麦区；A6是北方冬麦区出现频率最高的带型模式，而西南秋播麦区中D出现的频率最高。ω-区的E、H和M几种模式是以前国内外未曾报道的。且初步确定，这些模式对品质性状具有正效应。至于γ区,A、B、D、E和F在各区均有出现，其中B和E在各区出现的频率都很高，在26.1-39.6％之间。相反，H 仅出现在黄淮特种麦区，J仅限于西南秋播麦区。对于β-区醇溶蛋白，B型模式在所有区中都相当高，而模式A仅存在于第三区.对于α-区，模式A在Ⅲ区而模式D在Ⅱ区出现的频率很高。1BL.1RS易位系在中国小麦品种中出现频率高达41.2％，在I, II和Ⅲ麦区的出现频率分别为 45.5、43.5和35.2%。各生态区模式的差异可能是品种适应不同生态条件和人为选择的结果，但这有待进一步证明。由于醇溶蛋白位点（Gli-1）与LMW-GS位点（Glu-3）紧密连锁，本结果可为下面确定普通小麦LMW-GS等位基因变异所用。 2. 利用Gli-1与Glu-3的紧密连锁，以228个小麦品种/系为材料，首次对中国小麦品种麦谷蛋白亚基的6个位点进行综合分析，研究小麦籽粒蛋白与品质性状间的关系，结果表明6个高分子量（HMW）和低分子量（LMW）麦谷蛋白位点对蛋白质含量的效应大小为，Glu-D1>Glu-B3>Glu-A1=Glu-B1> Glu-A3=Glu-D3；对GMP含量的效应大小为, Glu-A3>Glu-B3>Glu-D1> Glu-B1>Glu-A1>Glu-D3；对湿面筋含量的效应大小为, Glu-B1>Glu-B3= Glu-D3>Glu-A3>Glu-A1>Glu-D1；对Zeleny沉降值的效应大小为, Glu-A1> Glu-B3>Glu-D3>Glu-D1>Glu-B1>Glu-A3；对SDS沉降值的效应大小为, Glu-B3>Glu-A1=Glu-D1=Glu-A3>Glu-D3>Glu-B1。对蛋白含量而言，各位点的最佳组合方式为1、17+18、5+10、Glu-A3e、Glu-B3g、Glu-D3b；对湿面筋含量而言，各位点的最佳组合方式为1、6+8、5+10、Glu-A3d、Glu-B3c、Glu-D3b；对Zeleny沉降值而言，各位点的最佳组合方式为N、17+18、5+10、Glu-A3d、Glu-B3d、Glu-D3b；对SDS沉降值而言，各位点的最佳组合方式为1、7+8、2.2+12、Glu-A3b、Glu-B3g、Glu-D3b。另外，分析了稀有亚基对5+12与2.2+12与品质性状的关系，认为5+12对品质有负效应，2.2+12对品质有正效应。在品质育种时，应对优异组合或优异亚基加以利用。 3. 首次利用重组自交系（RILs）为材料，研究麦谷蛋白亚基表达量与品质性状的关系，通过对重组自交系中各HMW-GS表达量的分析，认为，就单个亚基的表达量而言，7亚基最高；其次为2亚基、5亚基、12亚基和10亚基；亚基9和1的表达量最小；N亚基不表达。对成对出现的亚基对而言，x型和y型亚基的总表达量2+12>5+10>7+9>17+18。就单个亚基与品质性状的关系而言，仅有10亚基的表达量与蛋白含量的相关性达5%的显著水平，2亚基的表达量与湿面筋含量呈负相关，显著水平也达5%，其余单个亚基对品质性状均无显著影响；就x型/y型亚基的比例来看，2/12和5/10对湿面筋含量都有显著的负效应；对某一位点等位基因控制的亚基表达总量来看，2+12对SDS沉降值有显著负效应。另外，本研究得出：2＋12的亚基对的负效应主要体现在2亚基上，且在同一位点上，x型亚基的表达量大于y型。所以推导稀有亚基组合2+10很可能也是劣质亚基。 4. 以 Glu-A1、Glu-B1、Glu-D1、Glu-B3和Glu-D3作为5个因素对99G45/京771和Pm97034/京771杂交后代的蛋白质含量和SDS沉降值进行多因素方差分析。结果表明，Glu-A1和Glu-D3对蛋白含量的加性效应达5％显著水平；Glu-D1 * Glu-D3对蛋白质含量的互作效应也达5%显著水平；其余位点的加性和互作效应对蛋白质含量的影响均不显著。对SDS 沉降值而言，Glu-D1的加性效应最大,贡献率为4.2 % ,达1 %显著水平,其次是Glu-B1位点,贡献率为3.3% ,达5%显著水平。其余位点对SDS 沉降值的加性和互作效应均未达5%显著水平。总体而言, 各位点对蛋白含量的效应大小为Glu-D3 > Glu-A1 > Glu-D1>Glu-B1>Glu-B3；对SDS沉降值的效应大小为Glu-D1>Glu-B1> Glu-D3>Glu-A1> Glu-B3。Glu-D1和Glu-D3位点上等位基因变异对蛋白含量有显著或极显著影响，含Glu-D1d和Glu-D3 GD、Glu-D3 JD基因的株系分别比含Glu-D1a和Glu-D3 PD基因的株系有较高的蛋白含量；在该遗传背景下，麦谷蛋白各基因位点对蛋白含量的效应大小依次排列为：Glu-A1位点1>N；Glu-B1位点7+9>17+18>14+15；Glu-D1位点5+10>2+12；Glu-B3位点GB>JB>PB；Glu-D3位点GB>JB>PB。对SDS沉降值的效应大小依次排列为：Glu-A1位点1>N；Glu-B1位点7+9=17+18>14+15；Glu-D1位点5+10>2+12；Glu-B3位点GB>JB>PB；Glu-D3位点GB>JB>PB。所以，对蛋白含量和SDS沉降值均较好的组合为1，7+9，5+10，GB，GD。 5. 因为GB和PB对品质的效应有显著差异，选取LMW-GS位点特异扩增引物对京771、99G45和Pm97034的Glu-B3位点进行扩增，结果得到三个不一样的扩增片段（Genebank号为DQ539657－DQ539659），得到的基因片段与Genebank中已报道的同类序列高度同源。通过克隆片段组成的分析，发现对Pm97034的序列较京771和99G45段少一个7氨基酸的重复单元，这可能是它较另外两个片段对面筋强度影响小的主要原因；另外，在99G45的序列中，124位处出现L（亮氨酸）代替P（脯氨酸），158位处出现了T（苏氨酸）代换M(蛋氨酸)，这可能是99G45Glu-B3位点序列对SDS沉降值的效应显著优于Pm97034的原因。 6．通过对RILs各位点同普通小麦品种（系）各位点与品质关系的比较，发现对SDS沉降值的效应，各位点在不同研究材料中是不同的，普通小麦中：Glu-B3>Glu-A1=Glu-D1=Glu-A3>Glu-D3>Glu-B1，RILs中：Glu-D1>Glu-B1> Glu-D3>Glu-A1> Glu-B3。利用重组自交系材料（完全排除了1BL/1RS易位干扰）所得到的结果与Gupta and MacRitchie (1994)所得结论一致。进一步证实了1BL/1RS易位对小麦品质的重要影响。对蛋白含量而言，普通小麦品种（系）中，Glu-D1>Glu-B3>Glu-A1=Glu-B1> Glu-A3=Glu-D3，RILs中，Glu-D3 > Glu-A1 > Glu-D1>Glu-B1>Glu-B3，和对SDS沉降值的效应一样，推断在非1BL/1RS易位的情况下，各位点对其效应应为Glu-D3 > Glu-A1 > Glu-D1>Glu-B1>Glu-B3。 对同一位点的等位基因而言，普通小麦和重组自交系中Glu-A1和Glu-D1上的等位基因对品质性状的贡献是一致的，但Glu-B1上的等位基因对SDS沉降值的贡献发生了变化，普通小麦中17+18>7+9，RILs中7+9>17+18，这可能也是1BL/1RS造成的。 Baking quality improved is one of the main object of wheat bread in China. The overall objective of the present studies was to increase the understanding about protein quality in wheat, i.e. to make it possible to improve the production of wheat with desired quality for different end-uses. With the analysis of gluten protein in RILs, 99G45/Jing 771 and Pm97034/Jing, and 228 wheat cultivars or lines in China, the correlations between glutenin compositions and protein content, glutenin macropolymer(GMP), wet gluten content, Zeleny sedimentation value and SDS sedimentation value contentand breadmaking quality were studied. Also a rapid and efficient detection method of geneticpolymorphism at Glu-B3 loci in wheat was established using polymerase chain reaction(PCR).The results obtained were as follows: 1. Cultivated Chinese wheat germplasm has been a valuable genetic resource in international plant breeding. Patterns of gliadin among cultivated Chinese accessions are unknown, despite the proven value and potential novelty. The objective of this work was to analyse the diversity within improved Chinese wheat germplasm. The electrophoretic banding patterns of gliadin in common wheat cultivars and advanced lines were determined by acid-polyacrylamide gel electrophoresis. For 148 leading commercial cultivars and promising advanced lines used in our study, 48 patterns were identified, 29 corresponding to ω-gliadin, 9 to γ-gliadin, 5 to β-gliadin and 5 to α-gliadin. The most frequent patterns were A6 in ω; B in γ; B in β and A in the region of α. 116 band types appeared in the148 samples: 94 accessions had unique gliadin types, and 22 gliadin types while not unique were found in 54 accessions. The gliadin patterns of Chinese wheat cultivars and lines greatly differed from the patterns of wheat lines from other countries. Three patterns, E, J, H, M, N and O in the ω-zone had not previously been reported. Three wheat zones，the Northern Winter Wheat Region, the Yellow and Huai Valley River valleys Winter Wheat Region and the Southwestern Winter Wheat Region，in China showed different frequencies in their gliadin patterns. This information can be used to monitor genetic diversity with Chinese wheat germplasm. 2. To analyse the relationship between the loci and characteristics quality, we utilized the 228 cultivars/lines. The results showed that : For protein content, Glu-D1 >Glu-B3>Glu-A1=Glu-B1>Glu-A3=Glu-D3. For GMP content, Glu-A3>Glu-B3 >Glu-D1>Glu-B1>Glu-A1>Glu-D3. For wet gluten content, Glu-B1>Glu-B3= Glu-D3>Glu-A3>Glu-A1>Glu-D1. For Zeleny sedimentation value, Glu-A1>Glu-B3> Glu-D3>Glu-D1>Glu-B1>Glu-A3, For SDS sedimentation value, Glu-B3>Glu-A1= Glu-D1= lu-A3>Glu-D3>Glu-B1。For protein content, the best combination of 6 loci is (1,17+18,5+10,Glu-A3e, Glu-B3g,Glu-D3b). For wet gluten content, the best combination of 6 loci is (1,6+8,5+10,Glu-A3d,Glu-B3c,Glu-D3b). For Zeleny sedimentation value, the best combination of 6 loci is (N,17+18,5+10,Glu-A3d, Glu-B3d, Glu-D3b). For SDS sedimentation value, the best combination of 6 loci is(7+8,2.2+12,Glu-A3b, Glu-B3g,Glu-D3b)。Additional, we analysed the relationship between the subunits 5+12 and 2.2+12, think that 5+12 was negative for quality, 2.2+12 is postive for quality. It should be effective utilized. 3. It’s the first time to utilize RILs to study the relationship between subunits expression quantity and characteristics quality. The results showed that: For single subunit, the expression quantity of 7 is the highest. Then the 2, 5, 12 and 10. The expression of subunit 9 and 1 is the lowest. Subunit N is not expressed. For subunits, the expression quantity of x type and y type are 2+12>5+10>7+9>17+18. The significant relation of 5% only showed between the expression quantity of subunit 10 and protein content. The relationship between expression quantity of others and characteristic quality was not significant. For x type/ytype, 2/12 and 5/10 is negative relation insignificant level. For the subunit(s) in a loci, Only 2+12 effect SDS sedimentation value negative in significant level. 4. With RILs 99G45/Jing 771 and Pm97034/Jing 771, we found that: The effective of Glu-A1, Glu-D3 and Glu-D1 * Glu-D3 for protein content is significant at 5% level. The effect of other loci for protein wre not significant. For SDS sedimentation value, the effect of Glu-D1is the highest, which contribution is 4.2 % .Then the Glu-B1, contribution is 3.3%. The effect of other loci for SDS sedimentationvalue were not significant. In total, for protein content: Glu-D3 > Glu-A1 > Glu-D1>Glu-B1>Glu-B3; for SDS sedimentationvalue: Glu-D1>Glu-B1> Glu-D3>Glu-A1>Glu-B3. The effect of alleles in Glu-D1 and Glu-D3 loci are significant at 1% or 5%. In Glu-A1, 1>N; Glu-B1, 7+9>17+18>14+15; Glu-D, 5+10>2+12; Glu-B3, GB>JB>PB; Glu-D3, GB>JB>PB. For SDS sedimentation, Glu-A1, 1>N; Glu-B1, 7+9=17+18>14+15; Glu-D1, 5+10>2+12; Glu-B3, GB>JB>PB; Glu-D3, GB>JB>PB. The best combinations for SDS sedimentation value is 1，7+9，5+10，GB，GD. 5. Because of the difference of GB and PB for SDS sedimentation value, we selected the specific primer for LMW-GS loci to amplified the Glu-B3 of Jing771, 99G45and Pm97034. We got 3 amplify fragment (Gene Bank accession number are DQ539657－DQ539659）. We found that the fragment of Pm97034 were deleted a repetitive 7 amino acid domain, which is perhaps the reason effect the gluten strength. Furthermore, in the position 124 of sequence 99G45, L has been replaced with P. Position 158, T replaced M, which may be the reason why the Glu-B3 locus of 99G45 is prefer to Pm97034 when refer to SDS sedimentation value. 6. Comparing the results of RILs and common wheat, we found that perhaps just the1BL/1RS made the difference of loci in different accession.

无介体双室微生物燃料电池产电性能初步研究

本文对无介体双室微生物燃料电池的产电性能进行了初步研究，并根据不同运行阶段产电性能的优劣，对其中微生物的差异性进行了比较分析。全文分为两个部分： 第一部分：以乙酸钠为阳极原料构建双室微生物燃料电池（MFC），研究不同阴极受体、外接电阻、乙酸钠浓度和pH等因素对电池产电性能的影响，研究结果表明：在500mL的阴阳极反应体系中，选用乙酸钠作为阳极底物，质量浓度为6.46 g/L， pH 7.0，接入500Ω外电阻，阴极电子受体选择高锰酸钾的情况下，微生物燃料电池产电性能最好，最大电功率密度达到294.72 mW/m2，库伦效率能达到25.87%。在确定最适外接电阻阻值的同时对MFC内阻进行测定，阻值为871.87Ω。 第二部分：微生物燃料电池运行中，比较以厌氧污泥作为接种源的第一阶段和只接入附着有大量微生物电极的第二阶段的产电性能，得出第二阶段产电性能优于第一阶段，最大电功率密度达到353.57mW/m2，比第一阶段提高58.85 mW/m2；库伦效率为39.35%，增幅达52％左右；针对微生物燃料电池运行过程中，底物CH3COONa可能存在其它的代谢途径，本实验进行了第二阶段产电性能与CH3COONa消耗率关系以及阳极液面上方气体成分和含量的研究，发现第二阶段50h前CH3COONa的大量消耗主要用于微生物的生长，在整个运行过程中，阳极液面上方含有CH4和CO2；对气体测定的同时还发现，振荡能增强电功率密度的输出；通过对电极上和污泥中微生物差异性分析得出，δ-变形菌纲、β-变形菌纲和拟杆菌门的菌种更适应微生物燃料电池的运行环境，能在电极上大量富集，提高电池的产电性能，只接入附着有大量微生物的电极能有效降低热袍菌纲的菌种数量，降低了CH3COONa的无为消耗，有效提高了电池的库伦效率。 Electricity production in the mediator-less two-chambered microbial fuel cell(MFC) was researched. Based on the result in the different operation phase in the MFC, the microbial diversity was analysed. The paper involved two parts: Part 1: A two-chambered microbial fuel cell (MFC) was constructed with high-concentration sodium acetate as fuel in the anode. The influence of different electron acceptors in the cathode, external resistance value, pH value and concentration of sodium acetate on electricity generation in MFC was investigated. The result showed that the maximum power density of 294.72 mW/m2 and the coulombic efficiency of 25.87% was achieved at sodium acetate concentration of 6.46 g/L, pH 7.0, external resistance 500Ωin the anode and when using potassium permanganate as electron acceptor in the cathode. While decided the value of resistor, we found that shaking has effect on electricity production in the MFC. Part 2: Comparing the electricity production in different operation phases when using anaerobic sludge as inoculum in the first phase and microbes in the anodic electrode as inoculum in the second phase, the result showed that electricity production in the second phase was more than that in the first phase, the maximum power density of 353.57 mW/m2 and the coulombic efficiency of 39.35% was achieved, 58.85 mW/m2 and 52％ more than that in the first phase, respectively. According to the fact that CH3COONa might be metabolized in other pathway in the running process in the MFC, we determining the relationship between electricity production and CH3COONa consumption, and the gas content in the anode, we found that CH3COONa was mainly used for microbe growth before 50h, and the anode contained CH4 and CO2. At the same time, we found that shaking could improve power density. The analysis on diversity of microbe in the anodic electrode and anaerobic sludge showed that δ-proteobacterium, β-proteobacterium and Bacteroidetes adapted themselves to the running environment of MFC. The anode could enrich them to improve the electricity production while reduced the quantity of Thermotogales, which were obligately anaerobic organotrophs with a fermentative metabolism, to increase the coulombic efficiency effectively.

碱法草浆造纸蒸煮黑液厌氧生物降解性及其主要微生物类群研究

造纸行业是造成我国水环境有机污染物的重要污染源之一，其水污染的特点是小厂多、草浆多、工艺落后、污染扩散面广、造成废 水排放量大，每年排放的废水量约39亿立方米，占全国工业废水排放量的1/6，其中有机污染物（以BOD5计）160万吨左右，约占全 国工业废水中有机污染物总量的1/4。尤以占全国制浆造纸行业90%以上的碱法草浆造纸厂的蒸煮黑液量大面广，除含有机物外，还 含有木质素、残碱、硫化物、氯化物等污染物，属于PH值高、色度深、难于治理的高浓度有机废水，对水体污染特别严重，各地要 求治理呼声很高，急待研究并尽快找出各种有效的治理途径。对于碱法草浆蒸煮，黑液高浓度废水的治理，有各种方法，根据国内 的研究进展和我们已有试验工作表明，最经济有效，具有实用价值，在生产上可获得成功是厌氧处理法。近10多年来，国外关于高 效厌氧处理技术研究进展迅速，并出现了多种多样的工艺设备，如高效厌氧生物反应器，并在实用化方面取得了很大成绩，建立了 生产性装置，达到了高负荷运行，效果良好。本试验是根据我们已有研究基础，针对我国国情，对小型制浆造纸厂水污染防治除了 开发碱回收及各种综合利用技术外，要特别加强废水(废液)实用技术研究的指导思想，本试验采用改进型的上流式厌氧污泥床反应 器，设计了两种试验方案，通过试验结果如下。1. 试验方案I—碱法草浆黑液酸化和厌氧发酵I号UASB反应器动态模型试验结果表 明：(1). 采用中温35℃±1℃高效厌氧反应器USAB内装有填料(陶粒)和三相分离器，具有保持高浓度生物量和防止污泥流失的特点 ，污泥浓度Vs 可达30%以上，因而具有高效、节能、产能、滞留期短的优点，当进水CODcr在7500-10000mg/l，HRT由7天缩短到3天 ，有机容积负荷在1.22gCODcr/l·d-3.43gCODcr/l·d时，CODcr平均去除率可达55%-45.5%，最高CODcr去除率可达60.2-63.5%， BOD5去除率可达75.9-83.2%,沼气容积产气率可达0.29-0.67l/l·d，每克CODcr转化为沼气产率达0.39-0.48l/gCODcr·d，CH4含量 65.8-75.5%。厌氧发酵出水再用化学法进行后处理脱除难降解的木质素，CODcr总去除率达80%以上。(2). 动态试验结果表明：采 用酸化—厌氧发酵处理黑液工艺合理，技术路线可行。2. 试验方案II—黑液用化学法(Hcl)去除木质素进行厌氧发酵，II号UASB反 应器动态模型试验结果表明：(1). 采用中温35℃±1℃高效厌氧反应器UASB(内有软填料)，当进水CODcr7000-13000mg/l左右，HRT 由6天缩短到1天，有机负荷由0.98gCODcr/l·d增加到11gCODcr/l·d时，COD平均去除率均可稳定在70-77%，BOD5去除率为87.3- 93.1%，沼气容积产气率0.21-2.6l/l·d，每克CODcr转化为沼气产率为0.39-0.48l/gCODcr·d，高的可达0.53l/gCODcr·d，转化 率较高，CH4含量63-70%。(2). 试验证明碱法草浆黑液物化预处理—厌氧发酵处理的技术路线也是可行的，工艺合理、效果较好。 在有条件的工厂可采用。3.厌氧发酵阶段几大类群微生物计数表明：(1). 当发酵工艺和运行处于相对稳定状态时，微生物群体的 组成也达到相对的稳定，各类微生物之间保持动态平衡关系。当产乙酸菌的数量为107-108个/ml时，产甲烷菌的数量为105-106 个/ml，当产乙酸菌数量为106-107个/ml时，产甲烷菌的数量为103-105个/ml。(2).稳态运行条件下，黑液预处理为甲烷发酵创造 了有利的生态环境，获得了较好的处理效果和较高的COD转化为沼气的产率0.39-0.48l/g·CODcr·d，反应器中形成较为稳定而数 量较下水污泥中高1-2个数量级的厌氧发酵微生物区系组成。这一结果为黑液厌氧发酵提供了微生物理论依据。Paper industry is one of the important pollution source of water environment in our country. Its character of water pollution is many small factories, much grass pulp, disadvantageous technique, large preading area of pullution. Its effluent makes up 1/6 of whole country's industry wastwater. Its organic pollutant accounts 1/4 of whole country's. Alkaline grass paper pulp effluent with pollutants such as ligoin, remaining alkali sulfide, chloride besides organic material, is a kind of high concentrate organic wastewater which has high PH walug, dark colour and is difficult in treatment. There is urgent require to find ways to treat the wastewater. There are different ways to treat alkaline paper grass pulp effluent. According to the research advances and our experiment work, the most economical and useful way is anaerobic degradation which was advanced quick in last ten years. In the control of waste water of small pulp paper mill, the study of wastewater utilization technology should be emphasized, besides alkaline retrieving and different kinds of comprehensive utilization technology. Our experiment used modified UASB(Upflow Anaerobic Sludge Blanket Reactor). Two kinds of plan were disgned. The results are lined below. 1. The first experiment plant-aciding black pulp effluent and methanogenic digestion. The dynamic model experiment results of I-UASB reactor showed: (1)The mesophilic(35℃±1℃)high effect UASB reactor having haydite and threee state seperation in it had the character of keeping high bioimass concentration and preventing losss of sludge. It had advantages of high effect, energe saving, energe prodcing and short HRT(Hydroulic retention time). When the influent COD was 7500-10000mg, HRT was shortened from 7 days to 3days, organic loading rate was 1.22g-3.43COD/l· d, the average COD removal efficiency was 55%-45%. The highest COD efficiency was 60.2-63.5%, BOD removal of 75.9 -83.4% was achieved. Biogass production rate were up to 0.29-0.67l/l·d. Biogass converted efficiency from every gram of COD could reach 0.39-0.48l/gCOD·d. Methane content was 65.0-75.5%. Chemical method was used to deplate lignin in anaerobic digestion effluent. Total COD removal efficiency could be more than 80%. (2)Using aciding annaerobic digestion to treat the black effluent was reseanable in technique and technology. 2. The second experiment plan-anaerobic digestion was used after the chemical method was used to deplate lignin in the black effluent. The result of dynamic experiment of II-UASB reactor showed: (1)High effect mesophilic (35℃±1℃)UASB reactor having soft slaffing in was used. When influent COD was about 7000-13000mg/l, HRT was shortened from 6 days to 1 day and organic loading rate was increased from 0.90 to 11g COD /l·d, average COD removal efficiency remained stable on 70-77%. BOD, removal efficiency was between 87.3-93.1%. Biogass production rate was 0.2-2.6l/l ·d .Biogass converted efficiency from a gram of COD was 0.39-0.481/gCOD·d with the high value of 0.53l/gCOD·d. Methane content was 63-70%. (2)The way that using physical, chemical Pre-treatment-anaerobic digestion to treat alkaline black effluent is feasible and can be used in some factories when the condition exists. 3. Counting of several class of microoganisms in anaerobic digestion stage showed: (1)As the disgestion was in stable motion, the compositon of microorganic colony could get relative stable. Dynamic balance was remaining among different kinds of microorganism such as methanogenic bacteria, Acidogenic bacteria, sulfate reducing bacteria, and heterotrophic bacteria. (2)Under stable motion, the pre-treatment of black effluent produced favourable eco-enviroment for methanegenic digestion. Good treatment effect and high biogass convertent efficiency from COD(0.39-0.48l/g·COD· d)were gotten. Some stable and high quantity(10-100times more than sewage sludge)microorganism colony were formed in the reactor. This result provided theoretical basis for anaerobic digestion of black effluent.

微生物法沥取重金属污泥中铬的研究

本文研究了两种微生物及其组合沥取、回收用微生物法治理电镀铬废水产生的铬污泥中的铬。铬污泥富含C、N、O，含铬量为13%， 经X-光电子能谱分析铬以三价态(氢氧化铬)存在。二种微生物分别从一酸性矿水和酸性污泥中分离筛选得到，经鉴定为硫杆菌属 (Thiobacillus Beijerinek)的两个不同种，一为氧化亚铁硫杆菌(Thiobacillu ferrooxidans, TF), 另一为氧化硫硫杆菌 (Thiobacillus thiooxidans, TT)。研究并比较了不同微生物对污泥中铬的沥取能力，结果表明，TT菌沥取铬效率最高。振荡、动 态淋滤、静置等沥取方式经过研究表明动态淋滤为最佳，室温条件下(15-20℃)，污泥浓度为20g/L时，总铬沥出率达60%时所需时 间：动态淋滤为48.5h，振荡和静置方式分别为91.22，81.6h。研究了不同温度、不同起始PH、不同污泥浓度及非成熟菌液对微生 物沥取能力的影响：(1) 沥取前期，温度对铬的沥出影响较大；微生物沥取反应基本属一级反应；温度与反应速率的关系基本符合 Arrhenius方程，但沥取后期这一特点并不突出。(2) 沥取液最适起始PH为菌液自然PH；PH值的人为改变将使铬的沥出大大降低。 (3) 污泥浓度与铬的沥出呈正相关，但浓度高于30g/L时，铬的沥出量不再增加。(4) 非成熟菌液沥出铬的能力较差，但沥取液中 微生物生长繁殖较为活跃。总结微生物沥取反应最佳沥取条件为：TT成熟菌液、污泥浓度10g/L、温度25-36℃、动态淋滤方式，此 时铬几乎可100%从污泥中沥出。经扫描电镜分析，沥取开始时，微生物紧密吸附于污泥颗粒表面上，表面紧密吸附为微生物发挥功 能提供了基础。微生物沥取污泥中铬的反应机理推测为：硫细菌代谢产硫酸或氧化Fe2+成Fe3+，利用酸，Fe3+ 及自身氧化酶系统 氧化污泥中Cr3+为Cr6+,Cr6+溶出结晶为CrO3。This paper has studied bioleaching and recovery of Chronium(Cr)from electroplating sludge by two consortum of bacteria and their combination, with sludge produced by microbiological process treating electroplating wastewater containing Cr as material. The share of Cr is 13% and its state is Cr (OH)3 in the sludge. One of the bacteria in the paper was isolated from acid sewage sludge and the other was from acid mineral water. The former was tested and determined as Thiobacillus ferroxidans(TF) and the latter was Thiobacillus thiooxidans(TT). Different microorganisms, responsible for the metal leaching activity, have great influence on the efficiency of leaching. The results showed that TT has biggest power. Experiments were conducted to examined effects of three different ways of leaching(Shaking, Down-leaching, Static-leaching). When temperature was in-door's (15-20℃)and concentration of the sludge was 20g/L, the bioleaching time required to reach 60% of Cr solubilization with the above three ways were 91.2, 48.5, 81.6h respectively. Down-leaching was proved to be the most efficient. The influence of different temperature, initial PH, concentration of the sludge and non-mature inoculum had been studied. The results obtained reveal that: (1) The variation of temperature is important during the time from initial to middle of leaching. The reaction of bioleaching belongs to first-order. The relation between the bioleaching rate constant(In k)and temerature can be expressed by Arrhenius function. (2) The fittest initial PH is the nature PH of mature inoculum. Any alteration with it could cause clearly negative effection. (3) The concentration of the sludge can make strong influence on the bioleaching efficiency. But when the concentration is above 30g/L, the increasing of Cr in the solution is little. (4) If non-mature inoculum acts as the bioleachin microorganism, little quantity of Cr would be gained from the sludge. But the micormass in the solution is very active. The results from electron microscope showed that microorganisms adhered to the surface of the sludge and the adherence was the first stage of the bioleaching. Some salts of Cr can be obtained afer the water of the bioleaching solution being evaporated. By analysing the results of experiment with X-Ray spectroscopy, the salt was identified as CrO3. The recovery rate of Cr is 78.4%.

有机碳对混合菌群亚硝酸盐氮氧化的影响

自养硝化过程在自然界氮素循环和污水处理系统脱氮过程中起着关键作用。因此，了解有机碳对硝化的影响和硝化菌与异养菌之间的竞争对微生物生态学和污水处理系统设计都很重要。目前对氨氧化到硝酸盐氮过程的研究文献很多，但对亚硝酸盐氧化过程在异养菌的存在下如何受到有机碳影响的研究甚少。本文从生理生化指标、基因组学、蛋白组学三方面考察了在实验室条件下有机碳（乙酸钠）对硝化细菌和异养菌组成的混合菌群的硝化性能、菌群结构及代谢功能的变化的影响。 全文分为两大部分： 第一部分为乙酸钠对游离态硝化混合菌群的硝化性能和菌群结构的短期影响。混合菌株先在自养条件下进行连续培养，两个月后硝化速率达到20 mg N/(L·d)；而后离心收集菌体进行批式实验。在批式反应器中，初始亚硝氮均为126mg N/ L，乙酸钠-C 与亚硝酸盐-N 的比分别为0，0.44，0.88，4.41，8.82。结果表明：在低C/N 比（0.44 和0.88）时，亚硝酸盐去除速率比C/N=0 下高，细菌呈现一次生长；而在高C/N 比（4.41 和8.82）时，出现连续的硝化反硝化，亚硝酸盐去除率仍比对照下高，细菌呈现二次生长。不同C/N 比下微生物群落明显不同，优势菌群从自养和寡营养细菌体系（包括亚硝酸盐氧化菌，拟杆菌门,α-变形菌纲,浮霉菌门和绿色非硫细菌下的一些菌株）过渡到异养和反硝化菌体系 （γ-变形菌纲的菌株尤其是反硝化菌Pseudomonas stutzeri 和P. nitroreducens 占主导）。 第二部分为乙酸钠对硝化混合菌群生物膜的硝化性能和菌群结构的长期影响。接种富集的硝化混合菌群于装有组合式填料的三角瓶中，于摇床中自养培养；两个月后填料上形成生物膜的硝化速率达到20 mg N/ (L·d)；而后进行长期实验，每12 小时更换混合营养培养基（亚硝氮约200 mg N/ L，C/N 比同上）。结果显示：相较于C/N 比=0 时的亚硝酸盐氧化反应来说，低C/N 比出现了部分的反硝化，而高C/N 比则是几乎完全的反硝化。与对照比，C/N=0.44 时亚硝酸盐氧化速率并未受乙酸钠的影响，反而上升了，但C/N=0.88 时亚硝酸盐氧化速率有所下降。菌群结构分析表明自养对照与混合营养下微生物群落的不同；PCR-DGGE未检测出混合营养下硝化杆菌的存在，而显示异养菌尤其是反硝化菌的大量存 在。荧光定量PCR 结果表明随C/N 比上升，硝化杆菌数量从2.42 × 104 下降到1.34× 103 16S rRNA gene copies/ ng DNA，反硝化菌由0 增加至2.51 × 104 nosZgene copies/ ng DNA。SDS-PAGE 的结果表明不同C/N 比下的蛋白组较为复杂且呈现一定的差异性。 有机碳对亚硝氮氧化及微生物群落的影响很复杂，本文分别讨论了对游离态和生物膜固定态两种状态的混合菌群相应的短期和长期影响研究。研究发现，有机碳并非一定带来硝化的负影响，如果控制在适当的C/N 比范围，有机碳是有利于亚硝氮氧化的。这些发现阐明了有机碳和硝化反硝化的关系，填补了硝化微生物生态学上的空白，对污水处理系统中减少异养菌的影响并提高氮去除率有一定理论指导意义。 Nitrification plays a key role in the biological removal of nitrogen in both nature and wastewater treatment plant (WWTP). So, understanding of the effect of organic carbon on nitrification and the competition between nitrifying bacteria and heterotrophic bacteria is important for both microbial ecology and WWTP design and operation. Despite the fact that the nitrification process of ammonia to nitrate has been extensively investigated, it is not known how the process of nitrite oxidization is affected by organic carbon when heterotrophic bacteria are present. By measuring different physiological and biochemical parameters, as well as using genomic DNA and proteome analysis, we investigated the influence of organic (acetate) on nitrite oxidizing performance, community structure and metabolic function of nitrite-oxidizing and heterotrophic bacteria under laboratory conditions. The dissertation involves two parts: Part one deals with the effect of organic matter on functional performance and bacterial community shift of nitrite-oxidizing and heterotrophic bacteria under suspended state. The bacteria were prepared in a continuous-flow stirred reactor under autotrophic condition; after two months, the nitrification rate of the culture reached about 20 mg N/ (L·d); then the bacteria were harvested for the next batch experiments. The initial concentrations of nitrite were 126 ± 6 mg N/ L in all flasks, and sodium acetate (C) to nitrite (N) ratios were 0, 0.44, 0.88, 4.41, and 8.82, respectively. The results showed that at low C/N ratios (0.44 or 0.88), the nitrite removal rate was higher than that obtained under autotrophic condition and the bacteria had single growth phase, while at high C/N ratios (4.41 or 8.82), continuous aerobic nitrification and denitrification occurred besides higher nitrite removal rates, and the bacteria had double growth phases. The community structure of total bacteria strikingly varied with the different C/N ratios; the dominant populations shifted from autotrophic and oligotrophic bacteria (NOB, and some strains of Bacteroidetes, Alphaproteobacteria, Actinobacteria, and green nonsulfur bacteria) to heterotrophic and denitrifying bacteria (strains of Gammaproteobacteria, especially Pseudomonas stutzeri and P. nitroreducens). Part two describes the influence of acetate on nitrite oxidizing performance, community structure and metabolic function of nitrite-oxidizing and heterotrophic bacteria in biofilms. Bacterial enrichments was transferred into flasks with polypropylene carriers and cultured under agitated and autotrophic condition. After two month, the biofilms grown on the carriers had a nitrification rate of about 20 mg N/ (L·h); then the biofilms were refreshed with mixotrophic medium (nitrite were 200 mg N/ L in all flasks, and C/N ratios was the same as above) every 12 h. the results show: normal nitrite oxidization reactions were performed when C/N = 0, but nitrite oxidization and partial denitrification occurred with low C/N ratios (0.44 or 0.88). At high C/N ratios (4.41 or 8.82), we mainly observed denitrification. In contrast to C/N = 0, the nitrite oxidization rate was unaffected when C/N = 0.44, but decreased with C/N = 0.88. The structure of bacterial communities varied significantly between autotrophic and mixotrophic conditions. Nitrobacter was hard to detect by PCR-DGGE while heterotrophs and especially denitrifiers were in the majority under mixotrophic conditions. Real-time PCR indicated that the Nitrobacter population decreased from 2.42 × 104 to 1.34 × 103 16S rRNA gene copies/ ng DNA, while the quantity of denitrifiers obviously increased from 0 to 2.51×104 nosZ gene copies/ ng DNA with an increasing C/N ratio. SDS-PAGE indicated the complexity of and a certain difference between the proteome of nitrite-oxidizing and heterotrophic bacteria at different C/N ratios. We conclude that the influence of organic matter on nitrite oxidation and the community structure of NOB and heterotrophic bacteria is complex. In this dissertation, we focused on how sodium acetate influenced the system both under suspended state and in biofilms. We observed that acetate did not necessarily have a negative impact on nitrification. Instead, an appropriate amount of acetate benefited both nitrite oxidization and denitrification. These findings provide a greater understanding about the relationship between organics and nitrification; they fill the gaps in the field of microbial ecology of nitrifying bacteria; they also provide insight into how to minimize the negative impact of heterotrophic bacteria and maximize the benefit of nitrogen removal in biological treatment systems.

投加溶胞菌群对剩余污泥消化影响的初步研究

活性污泥法是目前世界上普遍应用的污水生物处理工艺，其在运行过程中产生大量的剩余污泥。由于剩余污泥处理费用巨大及污泥最终处置对环境具有潜在危害问题，污泥的处理和处置已经成为水处理领域关注的焦点。本文利用实验室筛选的溶胞菌群，在好氧消化的同时对污泥进行前处理，促进剩余污泥的破解与溶胞，再通过两相厌氧处理对污泥进行进一步消化，以研究投加溶胞菌对剩余污泥消化的影响。 本研究中溶胞菌污泥减量化技术分为两个部分，第一，污泥在溶胞菌作用下的好氧消化与污泥传统好氧消化的对比研究，利用取自成都三瓦窑污水处理厂剩余污泥，向好氧污泥消化反应器中投加溶胞菌，检测各项污泥指标，并通过同传统好氧污泥消化对比，以研究溶胞菌对污泥好氧消化的影响。第二， 经过溶胞菌处理后好氧消化的剩余污泥进行两相厌氧处理研究。通过建立好氧溶胞联合两相厌氧消化系统的来处理剩余污泥，并与相同条件运行的两相厌氧消化系统做对比，检测运行过程中系统中物质成分变化，研究了其处理能力和运行稳定性，探索了两相厌氧消化系统中的发酵类型差别，验证了好氧溶胞对剩余污泥的破解效果。 研究结果表明：污泥在溶胞菌作用下的好氧消化效果和消化效率均优于传统好氧消化。在溶胞菌群存在的情况下，剩余污泥的TSS和VSS去除率达到40%和53%，远高于传统好氧消化的12%和20%。污泥经过溶胞及好氧消化后，TCOD去除率达到54.4%。经过溶胞菌处理后的剩余污泥再进入两相厌氧处理系统，进入厌氧处理系统的剩余污泥的VSS／TSS比值约为0.62。在两相厌氧处理水力停留时间(HRT)为8d时，溶胞处理污泥厌氧消化后VSS去除率达到55.17%，对照组两相厌氧系统的VSS去除率平均值为18.53%。经过溶胞处理的两相厌氧系统的污泥减量了能力远高于对照组。两相厌氧系统的pH值和碱度说明系统运行较为稳定。产酸相的有机酸中乙酸含量高于丙酸和丁酸，说明发酵末端产物以乙酸为主。在20天的试验周期内，污泥溶胞处理后、两相厌氧系统产甲烷相产气量累积产气量为1.2L，对照组只有375ml。气体中甲烷含量都在55%左右。该研究结果表明，好氧溶胞对污泥有破解能力，溶胞处理对两相厌氧中产酸相水解污泥细胞有明显的促进作用，提高了产酸相的水解酸化能力和效率。该研究对于利用生物溶胞途径提高污泥消化效率具有重要意义。 The actived sludge process has been used more and more extensively, but the procedure will lead to a large quantity of excess sludge. The treatment of Excess activated sludge has becomes a focuses not only for it is a seriously negative effect on environment but also for the costly disposal comes subsequently. The cell lysing bacterium was keeped in our lab to joined in the digestion of the excess activated sludge which was carrying at the same time with pre-processing of sludge to investigated the influence of cell lysing bacterium on excess sludge. There are two part in the method of cell lysing bacterium digesting sludge technology, the first, comparison of excess sludge digestion between anaerobic Cell-lysing Pretreatment and Conventional Aerobic Process. The sludge which was collected from San Wanyao disposal plant in Chengdu was thrown into the aerobic process system with cell-lysing bacterium, then, the indexes were detected to compare the difference between the cell-lysing bacterium in aerobic process and the traditional method to determine the influence of cell-lysing bacterium on aerobic process ; The second, the research on the sludge which was pro-treated with cell-lysing and aerobic digestion in the diphase of anaerobic digestion system. The system of cell-lysing combined with diphase of anaerobic digesting was created to compare to the diphase of anaerobic digested system, the changes of mass constituent was detected to study the ability and steady of disposal. Moreover, the research explored the difference among the types of fermentation. The efficacious of aerobic process was been proved. The result shows that the digesting rate of aerobic process with cell-lysing bacterium was higher than the traditional process. The ratio of sludge is reach to 40%~53%, which was far more effectively than the traditional process rate of 12%~20%. The TCOD of sludge which was treated with cell lysing bacterium and Aerobic Process is reach to 54.4%. Then, the sludge was thrown into the diphase of anaerobic digesting system. VSS／TSS of sludge is 0.62, HRT is 6d, the reduction of VSS is reach to 40.8%. The pH and alkalinity indicate the steady running of the diphase anaerobic digest system. In the acerbity phasing, the content of acetic acid was more than butanoic acid and propanoic acid in organic acid, it is demonstrated that the main composition of final production of fermentation was Acetic Acid. During the 20d of experiment, methylhydride phasing of diphase anaerobic digest system produced 1.2L methylhydride, however, there is only 375ml in CK, the content of methylhydride in all gas phase was around the rate of 55%. The average ratio of VSS was 18.53% in CK diphase anaerobic digest system which was far more unavailable than the mass sludge rate of 55.17%. Results demonstrated that aerobic cell-lysing digested the sludge, the treat of cell-lysing could obviously promoted the hydrolyzeing of sludge cell in the acerbity phasing, which improved the ability and rate of hydrolization and acidification. This study is significant in inhenceing the rate of sludge digestion in the method of cell-lysing bacterium.

制革废水生物强化处理及中水回用试验研究

制革行业是轻工行业中仅次于造纸业的高耗水、重污染行业，作为劳动密集型行业，在解决大量人口就业问题的同时，也对所在地区环境造成了严重污染。目前我国制革行业每年排放废水8,000~12,000万吨，废水中含铬约3,500 t，SS为1.2×105 t，COD为1.8×105 t，BOD为7×104 t，对水体污染严重。 本研究在对厌氧酸化工艺进行研究、一级好氧处理段进行工艺比选研究的基础上，获得了匀质调节—SBBR—BAF的生物处理工艺，并依托该工艺进行了生物强化处理的研究，考察了菌剂的强化运行效果及其处理水回用的可行性。 研究表明，在进水COD＞3,000 mg/L，厌氧酸化具有很好的抗冲击作用，保证了好氧工艺出水COD＜200 mg/L；在进水COD＜3,000 mg/L，可只通过好氧处理实现出水COD＜200 mg/L。厌氧酸化停留时间选择不当，会导致厌氧出水硫化物浓度升高，严重影响好氧系统，会使好氧活性污泥因中毒而解絮。 研究表明，当进水COD为2,000~2,500 mg/L，NH4+-N为130~146 mg/L时，COD、NH4+-N去除率SBBR分别为93.8%~96.6%和14.5%~55.9%，SBR分别为88.8%~94.9%和13%~50.7%，表明SBBR优于SBR。同时，研究发现SBBR污泥增长率为0.05 kgVSS/kgCOD，仅为SBR0.57 kgVSS/kgCOD的8.8%。此外，研究发现SBBR在停止运行后经3个运行周期可回复原油能力，而SBR池经9个周期培养也不能恢复，说明SBBR恢复能力明显优于SBR。 研究表明，以匀质调节—SBBR—BAF为主的制革废水处理工艺，出水水质稳定，进水COD 801~2,834 mg/L、NH4+-N 87~203 mg/L，出水COD＜80 mg/L、NH4+-N＜10 mg/L，基本达到中水回用标准；操作简单灵活，没有污泥回流系统，污泥产率低，污泥处理费用低；工艺基本不需要添加化学药剂，既节约成本、又避免了二次污染；两级生物膜使得该工艺具有很强的耐冲击负荷能力，特别适合制革废水水质水量波动大的特点。 研究表明，高效菌对系统的启动具有一定的促进作用，强化系统生物膜6天可以成熟，对照系统生物膜9天可以成熟。同时高效菌能加速COD降解，缩短停留时间，强化系统6~8 h可使COD＜200 mg/L，对照系统8~10 h可使COD＜200 mg/L。长期运行表明，强化系统的SBBR在COD和NH4+-N的去除率都优于对照系统的SBBR。最终出水COD强化系统平均为53 mg/L、对照系统为74 mg/L。在模拟循环过程中，强化系统均有更高的稳定性。可实现8次理论循环，而对照系统只能实现4次理论循环。 研究表明，通过合理的工艺设计，可以实现猪皮制革废水达到《污水综合排放标准GB8976-1996》一级标准，同时满足工厂部分用水要求。通过添加高效微生物，可提高生物处理系统处理能力，使处理水能够满足工厂的多次回用。 As a labour-intensive industry, tanning has created large amount of working opportunities as well as caused severe contamination to environment. And it is one of the highest water-consuming and polluting industry, only second to manufacturing. At present time, Chinese leather industry emits wastewater about 80,000,000~120,000,000 t annually, which contains chromium about 3,500 t, SS 1.2×105 t, COD 1.8×105 t, BOD 7×104 t and ambient riverhead has been polluted greatly. Based on the research of anaerobic acidification and comparison of SBBR and SBR, biotreatment process (Homogenization—SBBR—BAF) had been established to amend the disadvantages of traditional sewage treatment such as too much sludge, high cost of advanced treatment and NH4+-N can not reach the emission standard. Research on the bioaugmentation was also been carried out. Researches showed, when COD of influent was beyond 3,000 mg/L, anaerobic acidification could resist strong impact, thus COD of effluent was less than 200 mg/L; when COD of influent was less than 3,000 mg/L, only throughout aerobic sewage treatment could COD of effluent beless than 200 mg/L. False residence tiome of anaerobic acidification would lead to the higher effluent concentration of sulfide and disintegration of aerobic activated sludge. Researches showed SBBR worked a better than SBR: when influent between 2,000 and 2,500 mg/L, NH4+-N between 130 mg/L and 146 mg/L, COD, NH4+-N removal rate of SBBR was 93.3%~96.6%, 14.5%~55.9% respectively while COD, NH4+-N removal rate of SBR was 88.8%~94.9%, 13%~50.7% respectively. Sludge growth rate of SBBR was 8.8% of that of 0.05 kgVSS/kgCOD. Besides, SBBR could recovered after 3 operating periods while SBR worked no better after 9 operating periods.Therefore, SBBR excelled SBR. Researches showed, effluent quantity of tannery wastewater treatment process (Homogenization—SBBR—BAF) was stable. When COD of influent was between 801 and 2,834 mg/L, NH4+-N was between 87 mg/L and 203 mg/L, COD of effluent was less than 80 mg/L, NH4+-N was less than 10 mg/L, which achieved the standard of reuse. This biotreatment was featured in low cost, easy and flexible management, less sludge, no inverse sludge system. Besides, this technique required no chemical, which could lower the cost and avoid secondary pollution. Great resistant of impact due to two membranes and was suitable for tannery wastewater which was featured by fluctuation of influent quality and quantity. Researches showed effective microorganisms promotes the startup of the process.Biofilm in the bioaugmentation process matured with 6 days while biofilm in normal process matured with 9 days. Effective microorganisms could accelerate the degradation of COD and shorten the residence time. Aggrandizement system could make COD<200 mg/L with 6 to8 hours while cntrolling system could make COD<200 mg/L with 8 to 10 hours. Long-term operating shows that SBBR in the bioaugmentation system worked better than the normal system in the treatment of COD and NH4+-N. The average COC of effluent in bioaugmentation system was 53 mg/L, normal system was 74 mg/L. In the simulative circulation process,aggrandizement process, which could fulfill 8 times theoretical circulation, works more stably than controlling process which could only fulfill 4 times theoretical circulation. Researches showed that reasonable design could make the wastewater meet the first grade of discharging standard of National Integrated Wastewater Discharge Standard (GB8976-1996), and partially meet the demand of water using of the factory. Adding effective microorganisms could enhance the biotreatment and make the effluents reuse many times.

Influence of iron promoter on catalytic properties of Rh-Mn-Li/SiO2 for CO hydrogenation

The effect of iron promoter on the catalytic properties of Rh-Mn-Li/SiO2 catalyst in the synthesis Of C-2 oxygenates from syngas was investigated by means of the following techniques: CO hydrogenation reaction, temperature-programmed reduction (TPR), temperature-programmed desorption and reaction of adsorbed CO (CO-TPD and TPSR) and pulse adsorption of CO. The results showed that the addition of iron promoter could improve the activity of the catalysts. Unexpectedly, the yield of C-2 oxygenates increased greatly from 331.6 up to 457.5 g/(kg h) when 0.05% Fe was added into Rh-Mn-Li/SiO2 catalyst, while no change in the selectivity to C-2 oxygenates was observed. However, the activity and selectivity Of C-2 oxygenates were greatly decreased if the Fe amount exceeded 1.0%. The existence of a little iron decreased the reducibility of Rh precursor, while the reduction of Fe component itself became easier. CO uptake decreased with increasing the quantity of Fe addition. This phenomenon was further confirmed by CO-TPD results. The CO-TPD and TPSR results showed that only the strongly adsorbed CO could be hydrogenated, while the weakly adsorbed CO was desorbed. We propose that Fe is highly dispersed and in close contact with Rh and Mn; such arrangements were responsible for the high yield Of C-2 oxygenates. (C) 2002 Elsevier Science B.V. All rights reserved.

ZnO clusters encapsulated inside micropores of zeolites studied by UV Raman and laser-induced luminescence spectroscopies

Using microporous zeolites as host, sub-nanometric ZnO clusters were prepared in the micropores of the host by the incipient wetness impregnation method. A small amount of sub-nanometric ZnO clusters were introduced into the channels of HZSM-5 zeolite, whereas a large quantity of sub-nanometric ZnO clusters can be accommodated in the supercages of HY zeolite and no macrocrystalline ZnO exists on the extra surface of the HY material. The vibrations of the zeolite framework and ZnO were characterized by UV Raman spectroscopy. The optical properties of these ZnO clusters were studied by UV-visible absorption spectroscopy and laser-induced luminescence spectroscopy. It is found that there are strong host-guest interactions between the framework oxygen atoms of zeolite and ZnO clusters influencing the motions of the framework oxygen atoms. The interaction may be the reason why ZnO clusters are stabilized in the pores of zeolites. Different from bulk ZnO materials, these sub-nanometric ZnO clusters exhibit their absorption onset below 265 nm and show a purple luminescence band (centered at 410-445 nm) that possesses high quantum efficiency and quantum size effect. This purple luminescence band most likely originates from the coordinatively unsaturated Zn sites in sub-nanometric ZnO clusters. On the other hand, the differences in the pore structure between HZSM-5 and HY zeolites cause the absorption edge and the purple luminescence band of ZnO clusters in ZnO/HZSM-5 show a red shift in comparison with those of ZnO clusters in ZnO/HY.

Natural 13C abundance as a tracer for studies of soil organic matter dynamics

A method for measuring the long- and medium-term turnover of soil organic matter is described. Its principle is based on the variations of 13C natural isotope abundance induced by the repeated cultivations of a plant with a high 13C/12C ratio (C4 photosynthetic pathway) on a soil which has never carried any such plant. The 13C/12C ratio in soil organic matter being about equal to the 13C/12C ratio of plant materials from which it is derived, changing the 13C content of the organic inputs to the soil (by altering vegetation from C3 type into C4 type) is equivalent to a true labelling in situ of the organic matter. Two cases of continuous corn cultivation (Zea mays: δ13C = −12%.) on soils whose initial organic matter average δ13C is −26%. were studied. The quantity of organic carbon originating from corn (that is the quantity which had turned-over since the beginning of continuous cultivation) was estimated using the 13C natural abundance data. After 13 yr, 22% of total organic carbon had turned-over, in the system studied. Particle size fractions coarser than 50μm on the one hand, and finer than 2μm on the other. contained the youngest organic matters. The turnover rate of silt-sized fractions was slower