橡胶的分子缠结和性能的研究

本工作由两部分组成：（I）以苯乙烯为溶剂时顺丁胶溶液缠结效应的研究；（II）稀土催化合成丁二烯与异戊二烯2二嵌段共聚物（PB－b-PI）的结构与性能的研究。以Ostward粘度计测定了不同胶样的Logη与LogC的关系曲线，研究临界缠结浓度C＊和在C＞C＊区域内的直线的斜率K与分子结构，温度和分子量分布间的依赖关系，用管子模型的GDE理论进行了分析。用TEM和WAXD方法研究了PB－b-PI二嵌段共聚物的低温结晶性能。在用TEM观察样品的结晶形态时，发现在所有共聚比范围内（PB／PI＝70／30～20／80），样品均具有结晶性。当用WAXD研究结温度和共聚组成对样品的结晶性能的影响时。发现：随着结晶湿度的降低和PB含量的增高，样品的结晶性能变好，结晶高增高。以单向拉伸研究了PB－b-PI和PB／PI共混物的力学性能。根据两相相分离的实验事实，我们提出了双缠结网络模型，其基本思想为：相分离使得两相分子各自形成缠结网络，由于两个缠结网络柔性的差异，外力使柔性大的PB缠结网络将首先产生，这种将随外力的增加而增加，当形变到某一定值时，PB缠结网络将自动产生松驰，通过嵌断点（或化学交联点），将力传递到刚性较大的PI缠结网络，使之产生形变，这种力的传递和一直到样品产生特大的形变而断裂。根据这一模型分析了有效缠结点，嵌段点和化学交联点在两相系中力的传递作用，并用Mooney-Rivlin方程给予了证明。上述模型成功地解释了本工作的实验现象。

间同立构聚丙烯的结晶行为和形态结构的研究

通过在不同温度下等温结晶,我们得到了sPP (ttgg)_2 螺旋构象的三种晶型(晶型I、晶型II和晶型III）。 晶型I和晶型II结构的晶体是亚稳态结构的晶体，在提高退火温度的条件下，能转变成稳定的晶型III结构的晶体。在室温下进行单轴拉伸时，无论是从玻璃态还是从螺旋构象的晶体，当拉伸比超过100%时，我们可以清楚地观察到(ttt)全反式构象晶体的生成。以上的实验结果在熔体取向薄膜的再拉伸中得到了证实。当由拉伸制得的(ttt)全反式构象晶体在不同温度下进行退火时，当退火温度超过46℃时，这种全反式构象的晶体能转变成(ttgg)_2螺旋构象的晶体。如果固定样品的长度进行退火时，这种转变直到退火温度超过100℃时才发生。提高退火温度有利于(ttgg)_2螺旋构象晶体的生成，而张力则有利于全反式构象的保持。根据已有的实验结果，我们提出了合理的转变机理。在不同温度下等温结晶，无论从玻璃态还是从熔体，我们都得到了不同形态结构的晶体，从球晶，孪晶到单层规则的长方形单晶，以及一种较少见球晶的生成过程，并分析了它的结晶机理。另外还研究了结晶温度对电子衍射中h10衍射条纹强度变化的影响。对比熔体结晶和玻璃态结晶，发现二者没有本质的差别，只是从玻璃态结晶所需的时间较长、结晶不够完善。这是由于处于玻璃态中的分子链活动能力较差的缘故。本文利用DSC等手段系统地研究了两种间同立构聚丙烯的结晶行为。利用Avrami方程作图求得 Avrami指数 n、结晶速率常数k;作图求得整体熔体结晶活化能ΔE；通过Hoffman-Weeks 和Thomson-Gibbs方程，作图求得平衡熔点T_m~o; 利用Hoffman和 Lauritzen的球晶生长速率方程作图求得成核常数K_g,发现 sPP1的结晶生长过程分成两个阶段，阶段III和阶段II，其K_g(III)/ K_g(II) = 1.9, 与理率基本吻合。同时也计算出折叠表面自由能σ_e和侧表面自由能σ以及链折叠功q。最后本文利用DSC分析了熔融双峰现象，研究了结晶速度，结晶温度和升温速率对熔融双峰的影响，提出了可能的解释。

真核生物FBPase和SBPase的进化研究

迄今为止，卡尔文循环是真核生物中唯一的二氧化碳固定途径。卡尔文循环利用NADPH和ATP将CO2转变成有机物，是自然界中有机物的主要来源。虽然，卡尔文循环中的酶促反应过程早在1956年就已经阐明，多数参与光合真核生物卡尔文循环的酶/基因的起源也有了较多的研究。但是，循环的5个关键酶中FBPase和SBPase的起源问题依然存在争议。本文首先对处于光合真核生物进化的关键地位的两种绿藻——衣藻和团藻中的FBPase进行了研究，进而对真核FBPase和SBPase进行了分子系统分析，以探讨光合真核生物中卡尔文循环的起源。 本研究发现：不同于一般光合真核生物中FBPase具有“胞质型”和“叶绿体型”两种亚型（分别参与糖异生途径和卡尔文循环），衣藻和团藻的基因组中只有一个编码叶绿体定位的FBPase （FBPase1）基因；多序列比对结果显示，FBPase1具有叶绿体型FBPase特有的参与光调节的氨基酸片段插入。再结合别人的“衣藻中的FBPase1的酶活性受光调节”的实验证据，本文认为该FBPase1为叶绿体型FBPase。有意思的是，通过搜索衣藻和团藻的基因组，本文发现了一个新型的FBPase酶基因（fbp2）。RT-PCR结果和EST数据均显示该基因在这两种绿藻中具转录活性。通过组装EST序列，获得了衣藻fbp2的cDNA和相应的蛋白序列。分析显示两种绿藻fbp2编码的氨基酸序列包含Li+-敏感磷酸酶基序（motif）和II类FBPase特有的FBPase_glpX结构域（domain）。这表明，该基因编码的蛋白（FBPase2）是II类FBPase。这是第一次在真核生物中鉴定得到这种原核型II类FBPase。分子系统分析进一步揭示了衣藻和团藻的共同祖先可能通过一次古老的水平基因转移事件，从放线杆菌亚纲的共同祖先中获得了该基因。由于放线杆菌亚纲II类FBPase具有胞质型FBPase的重要特征，因此推测所发现的FBPase2具有胞质型FBPase的特征。软件预测该FBPase2具有约20aa的信号肽，为叶绿体定位。再加上有研究表明衣藻的糖异生途径主要发生在叶绿体中。因此本文认为衣藻和团藻中也有两个FBPase同功酶；但与其它光合真核生物不同的是，胞质型FBPase发生了丢失，取而代之的是原核型II类FBPase参与其叶绿体中的糖异生途径。这种FBPase的不同情形很可能与这两种绿藻中独特的代谢途径区室化有关。 I 在细菌中，果糖-1，6-二磷酸和景天庚酮糖-1，7-二磷酸的去磷酸化是由果糖-1，6-/景天庚酮糖-1，7-二磷酸酶（F/SBPase）双功能酶催化的；但是，在光合真核生物中却是由底物特异的叶绿体型FBPase和SBPase分别催化的。通过结构域分析，本文发现细菌F/SBPase双功能酶可以划分为进化关系很远的两类（I类和II类）。通过选取来自更多细菌类群的代表序列，与真核FBPase和SBPase一起进行分子系统分析。结果显示，FBPase和SBPase既不是起源于I类F/SBPase双功能酶，也不是起源于II类F/SBPase双功能酶；而是分别起源于不同的真细菌I类FBPase。真核FBPase并没有与α-变形菌或蓝细菌FBPase聚在一起，却与不同类群细菌FBPase形成的clade形成姐妹枝。因此，尚不能明确真核FBPase起源于那类真细菌。真核SBPase形成的clade与ε-变形菌FBPase形成的clade在一起形成姐妹枝，表明SBPase很可能是通过一种未知机制起源于ε-变形菌的FBPase。 最后，基于上述研究结果并结合其它研究事实，本文还对光合真核生物中卡尔文循环的起源进行了探讨。

核仁蛋白基因组的原核起源与核仁蛋白网络进化组装分析

核仁是真核细胞间期核中最明显的结构，其主要功能是作为rDNA 的转录加 工和核糖体亚基合成、组装的场所。这一重要功能是包括原核生物和真核生物在 内的所有细胞生物都具有的，但核仁这一结构却只存在于真核细胞中。那么在从 原核到真核的进化过程中，核仁是如何进化而来，核仁的蛋白成分又是通过怎样 的方式从简单到复杂地组装起来，这些问题到目前为止都还没有得到深入的了 解。随着 “组学”的发展，核仁蛋白基因组的研究逐渐开展起来。越来越多的 核仁蛋白基因组数据为我们从“组学”的角度来探讨核仁的起源及其蛋白网络的 进化组装等问题奠定了基础。 1）本文首先通过比较目前仅有的三种真核生物（人，拟南芥，芽殖酵母） 的核仁蛋白基因组数据库，将三个核仁蛋白基因组共享的直系同源蛋白簇提取出 来构建出真核生物基本核仁蛋白基因组I (EBNP I)。由于拟南芥和芽殖酵母核仁 蛋白数据库明显偏小，为避免它们可能不全而带来的影响，进一步将人的核仁蛋 白基因组与拟南芥和芽殖酵母的全蛋白基因组共享的直系同源蛋白簇提取出来 构建出真核生物基本核仁蛋白基因组II（EBNP II）。EBNP 中的人核仁蛋白质序 列用作搜索序列去BLASTP 原核基因组，并构建同源关系矩阵进行聚类分析。 结果发现核仁蛋白基因组原核起源部分具有复合起源的特性，一部分来源于古细 菌，一部分来源于真细菌。并对EBNP II 中人核仁蛋白进行功能划分和对不同的 功能类群进行了起源分析，结果发现‘与核糖体有关’这一功能类群起源于古细 菌，‘与mRNA 有关’和‘与翻译有关’功能类群可能起源于古细菌，并在真核 阶段招募了大量蛋白成分，‘与染色质有关’这一功能类群可能起源于真细菌， 随后在真核阶段此功能类群招募了大量蛋白成分。 2）本文利用芽殖酵母的蛋白质相互作用数据，通过在原核生物基因组和原 生生物基因组的同源搜索，将芽殖酵母核仁蛋白分成不同进化时期产生的三个部 分：原核起源蛋白，原生生物起源蛋白，酵母特异蛋白。通过分析各部分蛋白质 的GO 注释，结果发现，原核起源蛋白主要行使与核糖体亚基合成与组装等核仁最基本的功能。通过比较各个部分之间以及各部分内蛋白质相互连接情况，结果 发现，芽殖酵母核仁蛋白网络按照不对称原则将招募的新蛋白组装入网络，整个 芽殖酵母核仁蛋白网络以原核起源蛋白所构建的框架为基础，原核起源蛋白起着 核心的作用。

竹叶青（Trimeresurus steinegeri）蛇毒凝血酶样酶研究

用MS-Sephadex C-25分离竹叶青粗毒，再经DEAE - Sephadex A-50和CM-Sephadex C-25纯化得到凝血酶样酶组分I。用DEAE-Sephadex A-50分离竹叶青粗毒，再经DEAE-Sepharose CL-6B纯化得到凝血酶样酶组分II。用聚丙烯酰胺凝胶电泳和SDS-聚丙烯酰胺凝胶电泳检查，组分I在两种电泳中均为一条带；组分II在两种电泳中均为二条带（靠得很近）。用聚丙烯酰胺凝胶电泳切割法证明了组分II的二条带都是凝血酶样酶。理化性质测定证明：组分I分子量为54，500，大约由261个氨基酸残基组成，其中Asp和Glu含量较高，含有14%的中性已糖，13.1%的已糖胺和14.7%的唾液酸，等电点为3.5，在230nm波长处有典型的蛋白质吸峰，在280nm处的消光系数为OD_(1cm)~(0.1%) = 0.855，组分I对温度对酸碱都有较好的稳定性。组分II用SDS-聚丙烯酰胺凝胶电泳测定，其分子量分别为54，000和47，000两种。凝胶电泳后用过碘酸-Schiff's试剂染色，证明组分II与组分I都是糖蛋白。

鞭毛虫尾藻（Oxyrrhis marina）细胞核的有丝分裂

尖尾藻细胞核的有丝分裂在涡鞭毛虫中间是非常独特的，可能对于了解有丝分裂的进化具有重要的意义。利用低速离心处理和低温（4℃）正常温度（19 ± 1℃）周期性自理相结合的办法，成功地实现了尖尾藻的同步化培养。以同步化培养的群体作为材料，用电子显微镜研究了尖尾藻细胞核有丝分裂过程中的超微结构变化。根据核中染色体的形态，间期细胞核可以分为两种类型-I型和II型。在II型间期核中，染色体比I型间期核中的染我体粗，其中的一部份呈棒状，另一部分形状不规则。在这两种类型的间期核中，细胞核和核仁都作园形。尖尾藻的分裂态Y形和U形染色体是出现在分裂间期细胞核中。前期时，核变成卵形，核仁伸长，在核膜上出现核小板（Nuclear Plaqes）；从核小板向核内发出微管。我们的观察表明在尖尾藻的核分开明过程中，虽然没有赤道板出现，但中期仍然是存在的。与许多高等真核生物一样，类尾藻的后期也可以分成后期A和后期B。后期A时，核内染色体向两极度移动，细胞核没有显著的伸长。染色体的极度向运动可能是由于与染色体相连的牵引微管的缩短和位于正在移向两极度的两条染色体之是的微管的伸长两种因素相互协调地起作用而引起的。在后期A细胞核中，位于两极度的微管组织者是多个位于核膜上的核小板，它们并未融合成为统一的微管组织中心。在后期B时，在细胞核的中间共很少见到微管，细胞核伸长，到末期开始时，核的长度大约是后期A细胞核的两倍。这种伸长很可能是由于中间区核膜的生长。在整个核分裂过程中，始终有核膜存在。我们发现微管也会与分裂中的核仁相连，这表明微管在核仁分裂中可能起着重要的作用。根据已有的资料，讨论了有丝分裂的机制和有丝分裂的进化。

森林管理对动物生境影响的模拟研究——以小兴安岭友好林业局为例

理解森林管理对野生动物生境的累积效应，需要对不同森林管理预案作出预测、分析和评价。本文，以小兴安岭友好林业局为研究区，利用空间直观景观模型LANDIS模拟其在6种森林管理预案—无采伐、皆伐、择伐I（禁伐区不采伐，限伐区和商品林区每10年采伐面积分别为5%和10%）、择伐II（限伐区和商品林区10%和15%）、造林I（采伐同择伐I，伐后人工更新）和造林II（采伐同择伐II，伐后人工更新）下200年内的森林景观变化。在查阅文献、野外考察和咨询相关专家的基础上，以GIS为平台，构建有着不同生境需求的3种动物：松鼠（Sciurus vulgaris）、马鹿（Cervus elaphus）和花尾榛鸡（Bonasa bonasia）在景观尺度上的生境适宜度指数（Habitat suitability index, HSI）模型，利用Arc的宏语言AML（Arc Macro Language）将LANDIS的输出结果与3种动物的HSI模型相结合，借助生境单元和类水平（Class level）上的景观指数（结合度指数等）评价动物生境，以反映不同森林管理预案对动物生境的影响。研究结果表明： 1）对于LANDIS的模拟结果而言，不同森林管理预案往往导致不同的森林演替动态：无采伐和2造林方案下，森林逐渐向成、过熟林演替，且造林方案显著增加了营林树种的分布面积；择伐I和择伐II下，由于成、过熟林被采伐，森林向后期演替的速度较为缓慢，而皆伐下，中、幼龄林的比重较大。定性而言，对于偏好早期演替的动物而言，皆伐下的生境适宜性较高；对于偏好中、后期演替森林的动物而言，2种择伐方案下生境适宜性较高；对于偏好后期演替的动物而言，无采伐和2造林方案下生境适宜性较高。 2）对于松鼠，无采伐和2造林预案下的生境单元和适宜生境（HSI≥0.5）结合度指数较高，适宜生境的平均斑块面积较大，表明这3种预案下的生境适宜性较高，而2择伐预案由于移去了大量成、过熟林，导致松鼠生境适宜性的下降，对于皆伐，尽管采伐量比2择伐预案的小，由于其不利于针叶林的天然更新，对松鼠的生境破坏更为严重。 3）对于马鹿，皆伐和2造林预案下的生境单元和结合度指数较高，而无采伐下的生境单元和结合度指数整体上又小于2择伐预案下的，这表明，适当的干扰反而利于提高马鹿的生境适宜性，因为，若没有这些采伐、造林等干扰活动，森林的郁闭度会随着树冠的封闭而逐渐加大，下层的许多不耐荫的树种就会消失或减少，降低马鹿的食物来源。 4）对于花尾榛鸡，若不考虑营林活动对其干扰，2造林预案下的生境单元和结合度指数较高，其次为无采伐预案，而皆伐和2择伐方案下较低，这表明，单纯的采伐的将减小花尾榛鸡的生境适宜性。 生态系统是一复杂的系统，因此，不可能存在适于所有动物的森林管理预案，对于管理预案的选择取决于管理者多方面（经济、生态和社会）的权衡。本文是将LANDIS与HSI相结合模拟评估森林管理预案对动物生境影响的一次尝试，研究结果表明了这一方法的实用性。尽管这一方法存在一定的局限性，但利用这一方法，能够可视化和量化结果，并比较不同森林管理预案，从生物多样性保护出发，为森林管理提供决策支持。最后，需要声明的是，本研究侧重方法的探讨，作为实际应用，还需要大量的野外工作来验证HSI模型。

长白山森林资源变化动态与景观格局预测

长白山区是东北地区重要的生态功能区，多年以来由于不合理的森林经营导致了森林质量明显下降，生态功能显著退化。因此，分析森林资源变化动态及其影响因素，明确人类活动对森林资源的影响，并提出合理的经营方案，对该区森林资源保护和恢复具有重要意义。本文以具有典型代表性的露水河林业局为研究对象，以1987，1995和2003三期森林资源二类调查资料为主要数据来源，利用ARCGIS、ARCVIEW和FRAGSTATS等软件为技术手段，分析该局资源变化、景观特征及格局变化及其与人为干扰的关系；同时以2003年森林资源二类调查数据为基础，应用景观空间直观模型LANDIS，设定5种不同的采伐预案：对照（无采伐）、方案I（一般公益林和商品林每十年分别采伐总面积的5%和10%+天然更新）、方案II（一般公益林和商品林每十年分别采伐总面积的10%和15%+天然更新）、方案Ⅲ（采伐同方案I+人工更新）和方案Ⅳ（采伐同方案II+人工更新），模拟未来300年森林景观变化，以揭示在不同的经营方案下森林资源的长期演替动态及空间分布特征，结果表明： 1.森林资源的质量发生了显著变化。过去16年间该区森林覆盖率增加了6.9%，但是森林蓄积量下降了60.58万m3；成、过熟林面积持续增加，中龄林面积急剧减少；1987年到1995年该区森林蓄积减少了110万m3，而1995年到2003年间森林蓄积增加了49.7万m3，这些表明1995年到2003年森林经营措施更有利于森林恢复。但中、幼龄林在16年间面积、蓄积持续减少，近年来采伐的主要是中龄林，说明成、过熟林面积的增加是以牺牲中龄林为代价的。 2.森林景观破碎化程度加剧。过去的16年间景观异质性程度加大，景观斑块内部的连通性增强。而景观类型的斑块数、边界密度、蔓延度逐渐增加，散布并列指数、多样性指数和均匀度等指数在逐渐的降低。1987年到2003年斑块数增加了979块，同时最大斑块占景观面积比例逐渐下降，1987年为28.7%，2003年减少到12.7%；由于经营方式变为采阔保针，大量采伐阔叶树种补植针叶树种，景观基质由1987年的阔叶混交林，转变为1995年后的混合景观基质；在斑块水平上阔叶混交林的平均斑块面积下降最多，边界分割程度最高，这表明天然次生林类型受人类干扰最大；从景观指数变化趋势看景观破碎化在1995年到2003年的变化趋势较1987年到1995年的变化趋势有所减缓。 3.LANDIS模型对5种不同的森林管理预案模拟结果表明：无采伐和低强度择伐+人工造林方案到模拟后期不同程度的增加了成、过熟林面积，更有利于增加森林质量和维护森林景观的完整性。人工造林与不造林相比，造林方案显著增加了造林树种的分布面积；造林方案下造林树种的分布面积、成、过熟林面积较其它预案要高，因此，用原始阔叶红松林中的关键种造林，有利于森林结构的优化及向阔叶混交林景观的转化，促进森林质量的提高和景观的恢复。

土壤优先水流及溶质先迁移的研究

本文以探讨土壤优先水流的特征和非吸附性离子优先先移为主要目的，通过对土壤优先水流穿透曲线的分析，定量的论证了优先水流的特征。在分析方法上进行了一系列尝试。得出了土壤优先水流的几个典型特征，即土壤优先水流的优先穿透、穿透曲线的不对称性、穿透曲线的拖尾和出流速率的不稳定性。在土壤优先水流特征分析的基础上，提出了判断土壤优先水流的定量方法。围绕非吸附性离子的优先迁移的实验结果，分析了阶跃输入和脉冲输入的非吸附性离子在不同水力负荷下的运移特点、优先水流的侧向入渗对土体中溶质分布的影响和耕作对土壤优先水流及溶质运移的影响。研究表明原状黑土、棕壤、暗棕壤和红壤中优先水流穿透时的孔隙容积分别为 0.05、0.051、0.048 和 0.042，穿透浓度为 0.759mg/l、0.087mg/l、0.115mg/l 和 0.117mg/l，在穿透时间上分别较对应的填充土柱提前 144、104、168 和 120 小时。原状土柱出流速率波动较大，其变异系数分别较对应的填充土柱高出 5～10 倍。在不同水力负荷下，原状土柱 Br 和 NO_3~- 的累积迁移量均明显高于填充土柱。不同水力负荷条件下，Br 和 NO_3~- 累积迁移量的差异不是来源于土壤优先水流，而是来源于土壤基质流。耕作可以影响土壤优先水流的穿透规律，并对土壤溶质运移构成影响。但耕作对土壤优先水流的穿透时间、穿透时的孔隙容积、穿透峰的出现和累积迁移量的影响并不遵从同一规律，这个过程不能通过孔隙的增减来简单描述的。优先水流存在下, Br 和 NO_3~- 在土壤中的分布表现为以大孔隙为中心向外径向递减，即 I 区溶质含量 ＞ II 区 ＞ III 区。土柱中溶质的径向分布以大孔隙为中心呈辐射状，表现距大孔隙越近溶质浓度越高。土壤优先水流可以引起非吸附性离子的快速大量迁移。这种迁移为深层次土壤和地下水中溶质浓度的大幅度提高提供了条件，一方面可能增加了深层次土壤污染修复的难度，一方面可以导致水体的富营养化，造成在下水污染的危险。

辽西地区油松纯林结构调整的研究

本文通过对辽西地区现有的47.1万hm~2油松林的广泛调查研究，探索出油松纯林结构调整的三种模式，即：人促更新调整模式（I）、镶边补空调整模式（II）和带状皆伐改造调整模式（III）。首次提出了辽西地区油松林结构调整全理林分油松所占的比例、皆伐带的宽度、镶嵌斑块的面积等数量指标。即当油松纯林集中连片的面积在10－15hm~2以上时，应进行调整；调整时，油松所占的最适比例为35％左右；在实行III模式调整时，最适带宽为12－30m，有时可加大到50m；实行II模式调整时，油松镶嵌斑块的最适面积为1－3hm~2左右，砍伐的油松斑块面积在 3hm~2以下。油松纯林经调整后，其林分的水源涵养能力、土壤肥力状况、林木生长量、抗虫效应、物种多样性等都有较大改善，尤其是固 N 树种的应用，效果更加明显。调整后的林分与油松纯林比可增各种效益达 7.4万元／hm~2·a。

EDMOCSS-有毒有机化学品在土壤－植物系统中暴露动态的新模型研究－－菲（Phenanthrene)在沈阳西部城市污水慢速渗滤土地处理系统的暴露动态模拟

有毒有机污染物（TOP）在土壤－植物系统暴露动态模型是TOP生态风险评价和管理的必要手段，是目前TOP环境科学研究的活跃领域。以往主要以经典的对流－扩散方程（CDE）为基础的建模途径存在诸多局限：i.基本过程机制和理论研究尚不完善，制约着系统模型的研究水平；ii.模型过于复杂，在应用上存在很大困难，；iii.普遍缺乏野外实际观测结果的检验。需要开辟和发展新的TOP在土壤－植物系统暴露动态模型正是在此方面的突破。EDMOCSS很大程度上克服了经典建模途径的局限，既包括几乎所有基本过程及其目前普遍接受和应用的理论，模型的结构又不很复杂，具有良好行为特征，应用上较方便。以优先有机污染物－菲（phenanthrene)为供试化学品的野外慢速土地处理系统（SR－LTS）小区动态观察结果，很好地验证了EDMOCSS核心模型－耕作层土壤分室模型的结果，表明了EDMOCSS有较好模拟预测能力，为EDMOCSS的深入研究和应用奠定了基础。

青藏高原东部混农季节性放牧及其对高山林线格局的影响

混农季节性放牧（agropastoral transhumance）通过作物种植和畜牧生产相结合的方式对不同海拔高度带上的资源进行相互补充利用，在亚洲兴都库什地区、青藏高原、横断山、东部及南部非洲、南美安第斯地区等具有悠久的历史。这种传统的生计系统几千年以来一直是居住在该地区的人类社会和自然生态系统相互作用的主要形式之一。这种传统的资源利用方式与山地自然植被以及特殊的山地人类文化和社会特征具有密切的协同演变关系。认识和理解这一关系，是山地生态学和人类学的核心科学问题之一。近年来，山地生态系统的多重功能性及动态演变对山区社会经济可持续发展的重要意义受到人们的不断关注。本文通过对云南省德钦县的12个自然村的混农季节性放牧以及对云南德钦、四川壤塘等山地植被格局特别是高海拔地带植被格局的的详细调查，探讨青藏高原东缘地区混农季节性放牧的主要特征、系统构成及相互关系，及其在全球变化、经济全球化和市场化及现代化过程中的变化趋势，分析混农季节性放牧与高山林线格局及生态系统的互动关系，旨在探讨山地地区人类活动与自然生态系统之间的互动关系，从而为山区社会经济可持续发展、环境建设和生物多样性保护等国家战略提供理论依据。 调查结果表明，混农季节性放牧是一种适应青藏高原东部高山峡谷地区环境因子及自然资源呈明显的垂直分布、资源数量稀少而时空分布异质性极高的生存环境的一种传统经济形式。这种传统的畜牧业的主要生产目的仍然是提供当地基本生存所需的产品，饲养牲口的种类和数量取决于农户的当地需求并且受资源的限制，因而维持在比较低的水平的。分布在不同海拔高度的放牧资源在一年中被牲口利用的时间也不同，互为补充，共同构成混农季节性放牧的资源基础。根据各社区永久居住点的位置和该村的土地资源特别是牧草地资源的分布范围，牲口迁移的距离和格局有较大的差异。。天然牧场仍然是最主要的畜牧业生产资源。混农季节性放牧中的农业系统和牧业系统互为补充，共同构成调查地区完整的的生计系统，农耕活动为放牧活动提供精饲料如粮食等和冬季饲料如秸秆， 其数量往往成为家庭畜牧业生产规模的主要决定因子之一。 通过对牲口数量和结构、牲口的时空迁移格局、牧业活动在整个经济活动中的相对重要性以及牧业活动和作物种植的关系方面的研究分析，混农季节性放牧在近几十年发生了深刻的变化。主要表现在牲口数量总体下降，牲口组成发生变化，牲口移动性降低、牧业活动的经济重要性下降以及牧业活动和种植活动之间的相互依存度降低等。上述变化的根本驱动力是发生在当地、地区及全球尺度上的环境、政治、社会经济、技术和文化等的变化，从而造成当地群众畜牧生产目标、土地利用和劳动力的分布等发生了变化。当地生计系统发生的改变可能会带来对方面而深刻的政治、社会经济、文化和生态影响。 混农季节性放牧这种古老的传统生计策略面临着许多挑战，如冬季饲料短缺、草场退化、缺乏市场竞争力、经济重要性降低、对年轻人缺乏吸引力、国家缺乏专门的政策指导等。与此同时，经济全球化、市场经济、新技术的应用、替代生计机会的增加、国家对于山地生态系统的作用的重新定位等也为传统生计系统转型、实现社会与生态共赢创造了机遇。 混农季节性放牧活动对亚高山及树线交错带生态系统系统的互动方式主要体现在以下几个方面：（1）牲口啃食、践踏等影响森林群落更新，改变森林群落的组成和结构，从而影响森林群落的演替进程和植被格局。林线边缘是搭建夏棚的首选地点，因此林线及树线交错地带就成了牲口活动的主要场所之一；（2）利用火烧开辟、维持和改良高山牧场； 3）在亚高山火灾迹地的放牧活动能够阻止火烧迹地的顺向演替； 4）牧民在林线附近获取建材和薪材等活动影响高山林线附近森林的结构和功能。 在调查区域，梅里雪山、白马雪山、甲午雪山的林线海拔高度在4200-4300m之间； 四川雅江、理塘一线，林线位置多在4300-4400m；四川壤塘二林场一带的林线主体在4100-4200m，在个别地区达到4300m； 在贡嘎山的南坡和东坡一带，林线位置在3600-3700m；而在四川松潘一带，林线位置主体在3700-3800米左右。树线高度的分布趋势和林线一致。混农季节性放牧及其有关人类利用活动使研究地区很多地方高山林线降低、树线交错带宽变窄或消失。在研究地区，总体情况是，阳坡和半阳坡（南坡、西南坡等）的林线和树线比阴坡和半阴坡（北坡、东北坡等）低，变化幅度达20-200m。这种差异主要是为了开辟牧场而人为清除了南向坡自然林线及其以上的植被从而使林线位置下降所致。在南坡自然林线保留得比较好的地方，林线和树线依然可以达到甚至超过北坡林线和树线的高度。放牧活动抑制了高山林线带火烧迹地的天然更新，从而使林线位置保持在目前的位置。 放牧活动对高山林线带森林群落更新的影响是显著的。自然林线内的乔木个体密度特别是新生苗和幼苗的密度大大高于非自然林线。没有放牧的自然林线及树线交错带内的I级个体（新生苗）密度达到725-2917株/公顷，而与之相对的处理样地内I级个体的密度只有0-228株/公顷；II级个体（高度10-50cm）也表现出类似的趋势，在没有放牧的自然林线及树线交错带样方内，其密度达到550-5208株/，而在放牧处理样方内只有14-321株/公顷。在非自然林线带样地内，在有正常放牧的样地内，完全缺乏I级个体。 从相对比例来看，没有放牧的样方内的I、II级个体在全部个体中所占的比例显著高于有放牧活动的样方。放牧使林线交错带的乔木幼苗数量显著减少，从而影响林线及树线交错带森林群落的天然更新过程。林线和树线交错带的灌木对乔木幼苗具有重要的保护作用，能够为树线树种如冷杉等幼苗的定居体提供有利的微气候环境，同时保护苗免受牲口的啃食和践踏。火烧以后接着进行放牧能够100%地抑制高山林线带的幼苗更新。 高山牧场放牧强度降低、使用时间缩短而低海拔地带放牧强度增加是研究地区混农季节性放牧系统的一个显著变化。这种变化也必然会引起各海拔带上的生态系统的变化。放牧强度的降低、生产性用火的停止将导致原来通过人工火烧而降低并通过进一步的火烧和放牧活动来维持的林线及其以上地带的灌木盖度和高度的增加，从而为林线森林群落的扩张创造条件。 青藏高原东部高山峡谷地区是我国重要的山地生态系统，在我国的生物多样性保护、生态环境建设、社会经济可持续发展战略中具有举足轻重的作用。正确认识人类特别是当地传统的生计系统与生态环境系统的互动关系是实现上述战略目标的前提。决策者必须以综合、系统的的视角协调促进社会经济可持续发展、保护生物及文化多样性和维持人、牲口和生态系统之间的平衡的多重目标。 Agropastoral transhumance, which makes a complementary exploitation of the natural resources at different altitudinal belts through a combination of migratory animal husbandry and crop cultivation, has a long history in Hindu-Kush Himalaya, Tibet Plateau, Hengduan Ranges, eastern and southern Africa and the Andes region of south America.For millennia, this traditional livelihood strategy has been one of the main forms of interaction between human societies inhabiting in these regions and their natural ecocystems. A close co-evolutionary relationship has been developed between this indigenous resources management systems and the mountain vegetation systems on the one hand and a unique set of cultural values and social features on the other. Understanding this relationship has been one of the core scientific issues in mountain ecology and anthropology. In recent years, the importance of the multiple functions of the mountain ecosystems and their dynamic changes in the sustainable socio-economic development of the mountain regions has gained increasing attention. This paper, which is based on a detailed study on the agropastoral practices of the 12 natural villages in Deqin County of Yunnan, and the mountainnn vegetation patterns in Deqin of Yunnan and Rangtang County of Sichuan, intends to reveal the major characteristics, system composition and the inter-relations of the subsystems of the agropastoral transhumance in Eastern Tibetan Plateau as well as the trends of changes of the system within the context of global changes, economic globalization and modernity process of China and analyze the relations between agropastoral transhumance and alpine ecosystem, ao as to understand the interactions between human activities and natural ecosystems of the mountains and provide theoretical basis for the national strategies in eocioeconomic development, environmental reconstruction and biodiversity conservation in the mountain regions. Results of the survey indicate that agropastoral transhumance in the investigated area is a traditional economic form that is highly adapted to the eastern Tibet Plateau where the topography features high peaks and deep gorges and where the highly variable environmental parameters and scanty natural resources exhibit a distinct vertical spectrum of distribution and great temporal and spatial heterogeneity. The main objective of pastoral management is still aimed at the production of basic goods and services of local people and thus the type and size of animals raised for each household mainly depend on local needs and are limited by the availability of natural resources. The scale of production is relatively low. Pastoral resources at different altidudinal belts are complementarily used at different seasons of a year and thus form the resources basis for agropastoral production of the study area. Migration distances and patterns vary with the location of the permanent settlements, the elevational distribution range of the resources of the villages concerned. Natural pastures (rangelands) are the main fodder resources and sumplement feedings only account for less than 5% of the total fodder consumption. Crop cultivation and pastoral activities support each other to form a complete livelihood system. The ability of the farmig lands (crop cultivation) to provide the pastoral activities with concentrates and sumplements often becomes a main factor limiting the scale of livestock production at household level. Agropastoral transhumance is experiencing drastic changes in recent decades as is reflected in the size and composition of animals, the seasonal migration pattern, the relative importance of pastoralism in the household economy and the interplays of agricultural and pastoral elements of the system. In general, there is a decline in animal population and mobility, a shift in animal composition to meet new needs arising from changed macro-economic situation, a decrease in the relative importance in the household economy and an increasing decoupling of agro&pastoral relations. The fundamental divers of these changes can be traced to environmental, social, economic, technological and cultural changes from local to global levels and such changes have further caused local changes in livestock management objectives, land use and distribution of labor forces. Changes in local livelihood systems could have profound political, socioeconomic, cultural and ecological conseuquences. Agropastoral transhumance, as an age-old traditional livelihood strategy, is facing multifacet challenges, such as winter fodder shortage, rangeland degradation, lack of market competitiveness, decrease in economic importance, lack of appreciation among the young generation and adequate policies from the government. At the same time, economic globalization, market economy, intrdoctution of new technologies, increase of alternative income generating opportunities and the national re-oreitation of policies on mountain ecosystems have all brought about new opportunities for the transformation of the traditional livelihood system and the synchronized development of local society and the environment. Agropastoral transhumance interacts with the ecosystems at the timberline and treeline ecotone mainly through the following aspects: 1)Animal browsing and stamping affect the regeneration process of the forest communities and alters the composition and structure of the forest which in turn affect the succession process and vegetation pattern of the forest communities. Forest edges are the priority locations for summer houses and therefore the timeline and treeline area becomes the major venues of aninal activities; （2）herders create, maintain and improve pastures through burning that remove the forest communities at the timeline and treeline ecotone; 3）immediate grazing on the fire sites can significantly prevent the fire sites from perogressive succession; and 4）herders harvesting of construction timber and firewoods affects the structure and functions of the forest communities at the timberline and treeline zone. Timberline position in the survey region shows geographical variations. It is around 4200-4300m in Meilixueshan, Baimaxueshan and Jiawuxueshan in Northwest of Yunnan and rises to 4300-4400m in Yajiang County and Litang County of Sichuan. In Rangtang of Sichuan, it is between 4100-4200m, though reaching 4300m in localized sites. In the southern and eastern slopes of Gongga Mountain, the timberline is only between 3600m and 3700m and in Songpan County at the upper reach of the Minjiang River the timberline is around 3700-3800m.Treeline pattern follows similar trend. In many places, agropastoral transhumance and related human activities have lowered the timberline and treeline and narrowed or removed the treeline ecotone. In the area of survey, generally speaking, timberlines and treelines are lower on the southern slopes than on the northern slopes, with a difference between 20 and 200m. This is mainly because that the use of fires to crerate pastures has removed the forest vegetation at the previous timberline and above. In fact, in many places, well-preserved forests on the south slopes have even high timberline position that the corresponding northern slopes. At subalpine zone, grazing activities could have prohibited the natural regeneration of many forest fire sites and maintained the forest position at the present level. Grazing has a significant impact on the regernation process of forest communities at the timberline zone. Natural timberline and treeline ecotone has much higher density of treeline species individuals especially the emergents and seedlings than the timberlines that are maintained by human activities. In natural timberline and treelien ecotone without grazing interference, the density of the I Class seedlings (less than 10cm in height) ranges 725-2917 /hm2; while that in the treatment plots (with grazing disturbance) is only 0-228//hm2；II Class seedlings (10-50cm）exhibit similar density trends, reaching 550-5208//hm2 in natural timberline without grazing but only 14-321//hm2 in the plots with grazing treatment. In the man-created timberlines, there is no I Class seedling at all in plots with normal grazing activities. In relative terms, in plots without grazing activities, the propotion of I Class and II Class seedlings is much higher than that in plots with grazing. Grazing activities have significantly reduced the number of seedlings in the timberline ane treeline ecotone, and thus affect the natural regeneration process of the forests. Shrubs at the timberline and treeline ecotone can effectively protect the seedlings from severe climate and animal tramping, thus increasing the survival rate of the seedlings. Grazing following fires can completely inhibit forest regeneration process at timberline. Changes in agropastoral transhumance will have great impact on the timberline and treeline pattern of the studied area. The decrease in grazing intensity on alpine pastrues and the cessation of the use of fires will result an increase in the cover and height of shrubs above the present human-maintained treeline, which will create further condition for the expansion of timberline forest communities. Eastern Tibet Plateau harbors some most important mountain ecosystems of China that are of vital importance to the country’s strategy in biodiversity conservation, environmental construction and sustainable sociaoeconomic development. A proper knowledge of the interactions between traditional livelihood systems and the ecosystems in the region is a precondition to the realization of the above strategic goals. Therefore, the decision-makers must have a holistic and systemic perspective so as to integrate the multiple objectives of promoting sustainable socioeconomic development, conserving biological and cultural diversity and maintaining the balances among people, animal population and the ecosystems.

增温对川西亚高山针叶林不同光环境下几种幼苗生长的影响

由于人类活动所引起的地球大气层中温室气体的富集已导致全球地表平均温度在20 世纪升高了0.6 ¡æ，并预测在本世纪将上升1.4-5.8 ¡æ。气候变暖对陆地植物和生态系统产生深远影响，并已成为全球变化研究的重要议题。位于青藏高原东部的川西亚高山针叶林是研究气候变暖对陆地生态系统影响的重要森林类型。森林采伐迹地和人工云杉林下作为目前该区人工造林和森林更新的两种重要生境，二者截然不同的光环境对亚高山针叶林不同物种更新及森林动态有非常重要的影响。 本文以青藏高原东部亚高山针叶林几种主要森林树种为研究对象，采用开顶式增温法（OTCs）模拟气候变暖来研究增温对生长在两种不同光环境下（全光条件和林下低光环境）的几种幼苗早期生长和生理的影响，旨在从更新角度探讨亚高山针叶林生态系统不同树种对气候变暖在形态或生理上的响应差异，其研究结果可在一定程度上为预测气候变暖对亚高山针叶林物种组成和演替动态提供科学依据，同时也可为未来林业生产管理者提供科学指导。 1、与框外对照相比，OTCs 框内微环境发生了一些变化。OTCs 框内与框外对照气温年平均值分别为5.72 ¡æ和5.21 ¡æ，而地表温度年平均值分别为5.34 ¡æ和5.04 ¡æ，OTCs 使气温和地表年平均温度分别提高了0.51 ¡æ和0.34 ¡æ；OTCs框内空气湿度年平均值约高于框外对照，二者分别为90.4 %和85.3 %。 2、增温促进了三种幼苗生长和生物量的积累，但增温效果与幼苗种类及所处的光环境有关。无论在全光或林下低光条件下，增温条件下云杉幼苗株高、地径、分支数、总生物量及组分生物量（根、茎、叶重）都显著地增加；增温仅在全光条件下使红桦幼苗株高、地径、总生物量及组分生物量（根、茎、叶重）等参数显著地增加，而在林下低光条件下增温对幼苗生长和生物量积累的影响效果不明显；冷杉幼苗生长对增温的响应则与红桦幼苗相反，增温仅在林下低光条件下对冷杉幼苗生长和形态的影响才有明显的促进作用。 增温对三种幼苗的生物量分配模式产生了不同的影响，并且这种影响也与幼苗所处的光环境有关。无论在全光或林下低光环境下，增温都促使云杉幼苗将更多的生物量分配到植物地下部分，从而导致幼苗在增温条件下有更高的R/S 比；增温仅在林下低光条件下促使冷杉幼苗将更多的生物量投入到植物叶部，从而使幼苗R/S 比显著地降低；增温在全光条件下对红桦幼苗生物量分配的影响趋势与冷杉幼苗在低光条件下相似，即增温在全光条件下促使红桦幼苗分配更多的生物量到植物同化部分—叶部。 3、增温对亚高山针叶林生态系统中三种幼苗气体交换和生理表现的影响总体表现为正效应（Positive），即增温促进了几种幼苗的生理活动及其表现：（i）无论在全光或林下低光环境下，增温使三种幼苗的光合色素含量都有所增加；（ii）增温在一定程度上提高了三种使幼苗的PSII 光系统效率（Fv/Fm），从而使幼苗具有更强的光合电子传递活性；增温在一定程度使三种幼苗潜在的热耗散能力（NPQ）都有所增强，从而提高幼苗防御光氧化的能力；（iii）从研究结果来看，增温通过增加光合色素含量和表观量子效率等参数而促进幼苗的光合作用过程。总体来说增温对幼苗生理过程的影响效果与幼苗种类及所处的光环境有关，增温仅在全光条件下对红桦幼苗光合过程的影响才有明显的效果，而冷杉幼苗则相反，增温仅在低光条件下才对幼苗的生理过程有显著的影响。 4、增温对三种幼苗的抗氧化酶系统产生了一定的影响。从总体来说，增温使几种幼苗活性氧含量及膜脂过氧化作用降低，从而在一定程度上减轻了该区低温对植物生长的消极影响；增温倾向表明使三种幼苗体内抗氧化酶活性和非酶促作用有所提高，从而有利于维持活性氧代谢平衡。但增温影响效果与幼苗种类所处的光环境及抗氧化酶种类有关，增温对冷杉幼苗抗氧化酶活性的影响仅在林下低光环境下效果明显，而对红桦幼苗抗氧化酶活性的影响仅在全光条件下才有明显的效果。 总之，增温促进了亚高山针叶林生态系统中三种幼苗的生长和生理表现，但幼苗生长和生理对增温的响应随植物种类及所处的光环境不同而变化，这种响应差可能异赋予了不同植物种类在未来气候变暖背景下面对不同环境条件时具有不同的适应力和竞争优势，从而对亚高山针叶林生态系统物种组成和森林动态产生潜在的影响。 Enrichment of atmospheric greenhouse gases resulted from human activities suchas fossil fuel burning and deforestation has increased global mean temperature by 0.6¡æ in the 20th century and is predicted to increase it by 1.4-5.8 ¡æ. The globalwarming will have profound, long-term impacts on terrestrial plants and ecosystems.The ecoologcial consequences arising from global warming have also become thevery important issuses of global change research. The subalpine coniferous forests inthe eastern Qinghai-Tibet Plateau provide a natural laboratory for the studying theeffects of climate warming on terrestrial ecosystems. The light environment differssignificantly between clear-outs and spruce plantations, which is particularlyimportant for plant regeneration and forest dynamics in the subalpine coniferous forests. In this paper, the short-term effects of two levels of air temperature (ambient andwarmed) and light (full light and ca. 10% of full light regimes) on the early growthand physiology of Picea asperata, Abies faxoniana and Betula albo-sinensis seedlingswas determined using open-top chambers (OTCs). The aim of the present study wasto understand the differences between tree species in their responses to experimentalwarming from the perspective of regeneration. Our results could provide insights intothe effects of climate warming on community composition and regeneration behavior for the subalpine coniferous forest ecosystem processes, and provide scientificdirection for the production and management under future climate change. 1. The OTCs manipulation slightly altered thermal conditions during the growingseason compared with the outside chambers. The annual mean air temperature andsoil surface temperature was 5.72 and 5.34 ¡æ (within the chambers), and 5.21 and5.04 ¡æ (outside the chambers), respectively. The OTCs manipulation increased airtemperature and soil surface temperature by 0.51 and 0.34 ¡æ on average, respectively.Air relative humidity was slightly higher inside the OTCs compared with the controlplots, with 90.4 and 85.3 %, respectively. 2. Warming generally stimulated the growth and biomass accumulation of thethree tree species, but the effects of warming on growth and development variedbetween light conditions and species. Irrespective of light regimes, warmingsignificantly increased plant height, root collar diameter, total biomass, componentbiomass (stem, foliar and root biomass) and the number of branches in P. asperataseedlings; For A. faxoniana seedlings, significant effects of warming on all the tested parameters (plant height, root collar diameter, total biomass, and component biomass) were found only under low light conditions; In contrast, the growth responses of B.albo-sinensis seedlings to warming were found only under full light conditions. Warming had pronounced effects on the pattern of carbon allocation. Irrespectiveof light regimes, the P. asperata seedlings allocated relatively more biomass to rootsin responses to warming, which led to a higher R/S. Significant effects of warming onbiomass allocation were only found for the A. faxoniana seedlings grown under lowlight conditions, with significantly increased in leaf mass ratio (LMR) and decreasedin R/S in responses to warming manipulation. The carbon allocation responses of B.albo-sinensis seedling to warming under full light conditions were similar with theresponse of A. faxoniana seedlings grown under low light conditions. Warmingsignificantly decreased root mass ratio (RMR), and increased leaf mass ratio (LMR)and shoot/root biomass ratio (S/R) for the B. albo-sinensis seedlings grown under full light conditions. 3. Warming generally had a beneficial effect on physiological processes of dominant tree species in subalpine coniferous forest ecosystems: (i) Warming markedincreased the concentrations of photosynthetic pigments in both tree species, but theeffects of warming on photosynthetic pigments were greater under low lightconditions than under full light conditions for the two conifers; (ii) Warming tended toenhance the efficiency of PSII in terms of increase in Fv/Fm, which was related tohigher chloroplast electron transport activity; and enhance non-radiative energydissipation in terms of in increase in NPQ, which may reflect an increased capacity inpreventing photooxidation; (iii) Warming may enhance photosynthesis and advancephysiological activity in plants by increasing photosynthetic pigment concentration,the efficiency of PSII and apparent quantum yield (Φ) etc. From the results, theeffects of warming on seedlings’ physiological performance varied between lightenvironment and species. The effects of warming on photosynthesis performance of B.albo-sinesis seedlings were pronounced only under full light conditions, while thephysiological responses of A. faxoniana seedlings to warming were found only underthe 60-year plantation. These results provided further support for the observationsabove on growth responses of seedlings to warming. 4. Warming had marked effects on antioxidative systems of the three seedlings.Warming generally decreased H2O2 accumulation and the rate of O2- production, andalleviated degree of lipid peroxidation in terms of decreased MDA content, whichalleviated to some extent the negative effects of low temperature on the plant growthand development in this region; Warming tended to increase the activities ofantioxidative enzymes and stimulate the role of non-enzymatic AOS scavenging,which helped to create an balance in maintaining AOS metabolites for the threeseedlings. Nevertheless, the effects of warming on antioxidative defense systems werepronounced only under the 60-year plantation for the A. faxoniana seedlings. Incontrast, the marked effects of warming on antioxidative defense systems for the B.albo-sinesis seedlings were found only under the full light conditions. In sum, warming is considered to be generally positive in terms of growth andphysiological process. However, the responses of growth and physiology performanceto warming manipulation varied between species and light regimes. Competitive and adaptive relationships between tree species may be altered as a result of responsedifferences to warming manipulation, which is one mechanism by which globalwarming will alter species composition and forest dynamics of subalpine coniferousforest ecosystems under future climate change.

同源四倍体水稻三系亲本的遗传差异、细胞遗传学比较研究及低直链淀粉含量突变体Wx基因序列分析

同源四倍体水稻（2N=4X=48,AAAA）是由二倍体水稻（2N=2X=24,AA）通过秋水仙素诱导染色体加倍后得到的新品系，具有优良的抗病性以及较高的蛋白质含量。因此，在四倍体水平上挖掘水稻的增产潜力成为水稻育种的新手段。同源四倍体水稻具有很强的遗传可塑性和很弱的遗传保守性，利用其作为水稻远缘杂交的桥梁，从野生物种中不断地引进有益的基因，这将有助于杂交水稻的多代利用和固定水稻的杂种优势。但是迄今为止，还没有关于同源四倍体水稻遗传多样性，遗传背景的报道。目前世界关于同源四倍体水稻的研究主要集中在中国，主要研究方向为培育、筛选结实正常的亲本材料，配置和筛选结实率正常或接近正常的组合。经过几十年研究，虽然在材料构建，细胞学研究等方面取得了较大进展，但同样由于结实率低的瓶颈问题未解决，而使多倍体水稻育种未能取得实质性进展。而近年来一些关于同源四倍体水稻低结实率机理的细胞学研究也由于缺乏统计学数据而缺乏说明性。本文用SSR标记，对选取的36个结实率正常同源四倍体水稻三系亲本和14个来源二倍体亲本，分析他们的遗传差异和群体遗传结构。本文还利用我们培育的高、低结实率的同源四倍体水稻恢复系、优良保持系和杂种F1及二倍体对照为材料，进行系统深入的细胞遗传学研究，进一步探讨同源四倍体水稻有性传递后代的发育过程，探索分裂期染色体行为特征与遗传性状稳定性的关系，为进一步选育多倍体水稻品种并将其应用于生产提供理论依据。同源四倍体水稻突变株D4063-1直链淀粉含量比来源二倍体明恢63下降一半,即其直链淀粉含量为5.23%，为研究其直链淀粉含量下降的原因，本文还根据普通水稻Wx基因设计引物，扩增测序获得了D4063-1Wx基因的全序列，与已报道Wx基因进行比对分析，并根据D4063-1和籼稻、粳稻的序列差异并根据D4063-1在该片段上的特征序列位点设计了用于识别D4063-1的寡核苷酸片段,为快速、准确的鉴别低直链淀粉的D4063-1创造了条件。 SSR标记具有基因组分布广泛、数量丰富、多态性高、容易检测、共显性、结果稳定可靠、实验重现性好、操作简单、经济、易于高通量分析等许多优点，被认为是用于遗传多样性、品种鉴定、物种的系统发育、亲缘关系及起源等研究的非常有效的分子标记。本研究选取了中国科学院成都生物所培育的同源四倍体和二倍体水稻亲本，并用36个微卫星标记进行了遗传差异和种群遗传结构分析。在50个品系中，我们观察到较高水平的多态性，每基因等位基因数（Ae）分布于2至6之间（平均值3.028），多态性信息含量（PIC）分布于0.04至0.76之间（平均值0.366）；期望杂合度（He）分布于0.04至0.76之间（平均值0.370），Shannon指数（I）分布于0.098至1.613之间（平均值0.649）。同源四倍体品系的等位基因数，期望杂合性和Shannon指数都比二倍体品系高。在供试50个品系中，较多材料均发现Rare基因，根据SSR多态性指数我们构建了同源四倍体和二倍体水稻的核心指纹库。F-统计值表明遗传差异主要存在于同源四倍体品系中（Fst=0.066）。聚类分析结果表明50个品系可以分为4个组。I组包括所有的同源四倍体和二倍体籼稻保持系，以及一个同源四倍体籼稻雄性不育系及其来源二倍体。II组仅包括IR来源的品系。III组比II组和IV组更复杂，包括同源四倍体和二倍体籼稻恢复系品系。IV组包括同源四倍体和二倍体粳稻品系。此外,由于等位基因及配子的遗传差异,同源四倍体与二倍体品系中存在单位点和双位点的遗传差异。分析结果表明,二倍体和四倍体水稻基因库的不同,其中遗传变异可以区分四倍体与二倍体水稻。同源四倍体水稻具有长期而独立的遗传性，我们能够选育并得到与二倍体亲本相比有特殊优良农艺性状的品系。 本研究以高结实率的同源四倍体水稻恢复系DTP-4、D明恢63及优良保持系D46B为材料进行农艺性状及细胞遗传学比较研究。DTP-4、D明恢63及保持系D46B的的染色体组成均为2N=4X=48，花粉母细胞具有较为理想的减数分裂行为，配对染色体的比率在99%以上，这与理论染色体组构成相符。DTP-4和D明恢63PMC减数分裂各个时期单价体和三价体的比例都非常低，而在MI， PMC观察到较多的二价体和四价体且四价体多以环状形式出现，其最大频率的染色体构型分别为12II 6IV和10II 7IV。恢复系DTP-4和D明恢63在MI四价体频率分别为2.00/PMC和2.26/PMC，而保持系D46B在MI四价体频率为6.00/PMC，极显著地高于恢复系品系，表明保持系D46B具有更好的染色体配对性质；AI保持系D46B的染色体滞后频率为10.62%，远低于恢复系材料DTP－4和D明恢63的19.44％和23.14％，接近二倍体对照明恢63的7.30%水平；TI保持系D46B具有比恢复系更低频率的微核数。而在TII，D46B的正常四分小孢子比率不但高于恢复系品系甚至高于二倍体对照。对高低结实率的同源四倍体水稻恢复系和杂种F1代的花粉育性，结实率和细胞遗传学行为进行了比较研究。DTP-4, D明恢63, D46A´DTP-4和D46A´D明恢63的花粉育性和结实率比D什香和D46A´D什香显著提高。减数分裂分析的结果表明，DTP-4，D明恢63，D什香，D46A´DTP-4，D46A´D明恢63和D46A´D什香其减数分裂染色体构型分别为：0.05I +19.96 II (9.89棒状+10.07环状) +0.01III + 2.20 IV, 0.11I +19.17 II (8.90 棒状+10.37 环状) +0.09III + 2.26 IV + 0.01 VI, 1.33I +9.46 II (4.50 棒状+4.96 环状) +0.44III + 6.02 IV + 0.09VI + 0.09 VIII, 0.02I +14.36 II (6.44 棒状+7.91 环状) +0.01III + 4.80IV + 0.01VIII, 0.06 I +17.67 II (11.01 棒状+6.67 环状) +0.06 III + 3.10 IV + 0.01 VI and 1.11 I +11.31 II (5.80 棒状+5.51 环状) +0.41 III + 5.63 IV+0.03VI+0.03VIII。在同源四倍体水稻恢复系和杂种F1代材料中，最常见的染色体构型为16II +4IV和12II +6IV。在减数分裂过程中，结实率较高的材料染色体异常现象较少而结实率较低的材料染色体异常现象较严重。在杂种F1代中，二价体的比例要低于其相应的恢复系亲本，同样的，单价体，三价体和多价体的比例相比其恢复系亲本也偏低。然而，在减数分裂MI，杂种F1代中四价体的比例要显著高于其恢复系亲本。在中期I，每细胞单价体的比例和花粉育性呈现出极高的负相关(-0.996)，当单价体数目升高时，花粉育性下降。其次是每细胞三价体的比例(-0.987)，之后则是每细胞多价体的比例与花粉育性的负相关(-0.948)。但是统计分析表明，二价体和四价体的比例对花粉育性和结实率没有显著影响。这一结果表明出了花粉育性和细胞减数分裂行为的相关性，同源四倍体的减数分裂行为为筛选高结实率的同源四倍体种系提供了理论依据。 突变体是遗传学研究的基本材料。利用突变体克隆水稻基因,并进而研究基因的生物学功能是水稻功能基因组学的重要研究内容。本课题组在多年的四倍体水稻育种研究中已获得多个低直链淀粉含量突变体，其中一些突变体在直链淀粉含量下降的同时，胚乳外观也发生了显著改变，呈半透明或不透明。同源四倍体水稻突变株D4063-1直链淀粉含量比来源二倍体明恢63下降一半,即其直链淀粉含量为5.23%。为研究其直链淀粉含量下降的原因，我们根据普通水稻Wx基因设计引物，扩增测序获得了D4063-1Wx基因的全序列，与已报道Wx基因进行比对分析；同源四倍体水稻D4063-1Wx基因最显著变化为在外显子序列中发生了碱基缺失，导致移码突变，在第9外显子终止密码子提前出现。D4063-1Wx基因碱基位点的变化还导致了其序列上的酶切位点的变化,对常用限制性内切酶位点分析分析结果表明同源四倍体水稻相对于籼稻和粳稻多了2个sph1酶切位点，相对于粳稻减少了6个Acc1，增加了4个Xba1，1个Xho1，1个Pst1和1个Sal1酶切位点。聚类分析表明D4063-1Wx基因序列与籼稻亲源关系较近，由此推测D4063-1Wx基因来源于籼稻的Wxa基因型。另外，根据D4063-1Wx基因的碱基差异，我们推测D4063-1Wx基因外显子碱基变化导致的RNA加工障碍是其直链淀粉降低的主要原因，并可能与其米饭较软等品质相关。本文还根据D4063-1和籼稻、粳稻的序列差异并根据D4063-1在该片段上的特征序列位点设计了用于识别D4063-1的寡核苷酸片段,并作为PCR反应的引物命名为AUT4063-1,将该引物与我们设计的扩增普通籼稻、粳稻的Wx基因引物F5配合使用建立了识别D4063-1的显性和共显性两种检测方式的分子标记,为快速、准确的鉴别低直链淀粉的D4063-1创造了条件。 研究同源四倍体水稻基因组的遗传差异，探索同源四倍体水稻的遗传规律，研究分裂期染色体行为特征与遗传性状稳定性的关系,旨在揭示四倍体水稻中同源染色体配对能力的遗传差异，为进一步选育多倍体水稻品种并将其应用于生产提供理论依据。 Autotetraploid rice (2N=4X=48, AAAA) is a new germplasm developed from diploid rice (2N=2X=24, AA) through chromosomes doubling with colchicines and is an excellent resource for desirable resistance genes to the pathogens and high protein content. Therefore, heterosis utilization on polyploidy is becoming a new strategy in rice breeding. At present, the main research on autotetraploid rice centralizes in China. Breeding effort has been made to improve autotetraploid rice genetically, however, the progresses are limited due to higher degree of divergence between hybrid sterility and polygenic nature. But to date, almost nothing is reported about the genetic diversity, original and genetic background of autotetraploid rice. Despite several reports on cytological analysis of the mechanisms of low seed set in autotetraploid rice still the results are inconclusive due to lack the statistical evaluation. Therefore, the study on the mechanisms of low seed set in autotetraploid is a priority for rice breeding. Microsatellites or simple sequence repeats (SSRs) are the widely used marker for estimating genetic diversity in many species, including wild, weedy, and cultivated rice. In our research, genetic diversity and population genetic structure of autotetraploid and diploid populations collected from Chengdu Institute of Biology, Chinese Academy of Sciences were studied based on 36 microsatellite loci. For the total of 50 varieties, a moderate to high level of genetic diversity was observed at population levels with the number of alleles per locus (Ae) ranging from 2 to 6 (mean 3.028) and PIC ranging from 0.04 to 0.76 (mean 0.366). The expected heterozygosity (He) varied from 0.04 to 0.76 with the mean of 0.370 and Shannon’s index (I) ranging from 0.098 to 1.613 (mean 0.649). The autotetraploid populations showed a slightly higher level of effective alleles, the expected heterozygosity and Shannon’s index than that of diploid populations. Rare alleles were observed at most of the SSR loci in one or more of the 50 accessions and core fingerprint database of the autotetraploid and diploid rice was constructed. The F-statistics showed that genetic variability mainly existed among autotetraploid populations rather than among diploid populations (Fst=0.066). Cluster analysis of the 50 accessions showed four major groups. Group I contained all of the autotetraploid and diploid indica maintainer lines and a autotetraploid and its original diploid indica male sterile lines. Groups II contained only original of IR accessions. Group III was more diverse than either group II or IV and comprised of both autotetraploid and diploid indica restoring lines. Group IV included japonica cluster of the autotetraploid and diploid rices. Furthermore, genetic differences at the single-locus and two-locus levels, as well as components due to allelic and gametic differentiation, were revealed between autotetraploid and diploid varieties. This analysis indicated that the gene pools of diploid and autotetraploid rice are somewhat dissimilar, which made a variation that distinguishes autotetraploid from diploid rices. Using this variation, we can breed new autotetraploid varieties with some new important agricultural characters but the diploid rice has not. Cytogenetic characteristics in restorer lines DTP-4, DMinghui63 and maintainer line D46B of autotetraploid rices were studied. DTP-4, DMinghui63 and D46B showed the advantage of high seed set and biological yield. The meiotic chromosome behavior was slightly irregular in DTP-4, DMinghui63 and D46B. We observed less univalent, trivalent and multivalent at MI, but more bivalent and quadrivalent were observed. The most frequent chromosome configurations were 12II 6IVand 10II 7IV in restorer and maintainer lines, respectively. The quadrivalent frequency of DTP-4 and Dminghui63 at metaphase(MI) was respectively 2.00/PMC and 2.26/PMC. However that frequency of D46B was 6.00/PMC, which was greatly significantly higher than DTP-4 and Dminghui63. That indicates the maintainer D46B has better chromosome pairing capability in metaphase (MI). The frequency of lagging chromosomes of the maintainer D46B at anaphaseI (AI) was 10.62%, which was significantly lower than that of DTP-4(19.44%) and Dminghui63(23.14%) and nearly reaching the level of diploid CK(7.30%). In telophaseI (TI) maintainer D46B showed lower frequency of microkernel at TI and lower frequency of abnormal spores at telophaseII(TII). We also studied pollen fertility, seed set and cytogenetic characteristics of restorer lines and F1 hybrids of autotetraploid rice. DTP-4, DMinghui63, D46A´DTP-4 and D46A´DMinghui63 showed significantly higher pollen fertility and seed set than DShixiang and D46A´DShixiang. Pairing configurations in PMC of DTP-4, DMinghui63, DShixiang, D46A´DTP-4, D46A´DMinghui63 and D46A´DShixiang were 0.05 I+19.96 II (9.89 rod+10.07 ring)+0.01 III+2.20 IV, 0.11 I+19.17 II (8.90 rod+10.37 ring)+0.09 III+2.26 IV+0.01 VI, 1.33 I+9.46 II (4.50 rod+4.96 ring)+0.44 III+6.02 IV+0.09 VI+0.09 VIII, 0.02 I+14.36 II (6.44 rod+7.91 ring)+0.01 III+4.80 IV+0.01V III, 0.06 I+17.67 II (11.01 rod+6.67 ring)+0.06 III+3.10 IV+0.01 VI and 1.11 I+11.31 II (5.80 rod+5.51 ring)+0.41 III+5.63 IV+0.03 VI+0.03 VIII, respectively. Configuration 16 II+4 IV and 12 II+6 IV occurred in the highest frequency among the autotetraploid restorers and hybrids. Meiotic chromosome behaviors were less abnormal in the tetraploids with high seed set than those with low seed set. The hybrids had fewer frequencies of bivalents, univalents, trivalents and multivalents than the restorers, but higher frequency of quatrivalents than the restorers at MI. The frequency of univalents at M1 had the most impact on pollen fertility and seed set, i.e., pollen fertility decreased with the increase of univalents. The secondary impact factors were trivalents and multivalents, and bivalents and quatrivalents had no effect on pollen fertility and seed set. The correlative relationship between pollen fertility and cytogenetic behaviors could be utilized to improve seed set in autotetraploidy breeding. The amylose content of autotetraploid indica mutant Rice D4063-1 dropped by half than diploid Minghui 63, that is, its amylose content of 5.23%.The whole sequence of Waxy gene of D4063-1 is amplified and sequenced. And the discrepancy of bases is found comparing to the reported Waxy gene. The Waxy gene of autotetraploid Rice D4063-1 had a base deletion in exon sequence, which resulted frameshift mutation in exon 9 and termination codon occur early. The mutation of Wx also led to the change of some common restriction endonuclease sites. Results showed compared to indica and japonica, D4063-1 had two adding sph1 sites. Compared to japonica, D4063-1 had six decreasing Acc1, a adding Xho1, Pst1 and Sal1 restriction sites. Phylogeny analysis shows that the DNA sequence of Waxy gene of D4063-1 is closer to Indica, and we suppose that the Waxy gene of D4063-1 is origin from genotype Wxa. In addition, according to the base differences of Wx in D4063-1, we deduce that RNA processing obstacle led by base change of intron is the main cause to low the amylose content, and related to phenotype of its soft rice. Based on analysis of fragments of D4063-1, indica and japonica and according to the special point of the three species, primers as markers-AUT4063-I were designed for distinguishing the D4063-1 from other rice. Combining with primer pair F5, dominant and codominant ways were established for discriminating them., rapid and correct identification of D4063-1 from other rice could be done. The genetic analysis is important to ensure the original of autotetraploid rice, for maintaining the “distinctiveness” of autotetraploid varieties, and to differentiate between the various genetic background of autotetraploid rice. The autotetraploid breeding will benefit from detailed analysis of genetic diversity in the germplasm collections. Further investigation on mechanisms of meiotic stability should benefit polyploid breeding. These findings demonstrated opportunity to improve meiotic abnormalities as well as grain fertilities in autotetraploid rice.

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.