水稻同源四倍体高代自交系农艺性状分析及遗传差异研究

课题组在不断地创制新的同源四倍体材料的同时，连续多年以提高结实率为目的培育、筛选自交系材料，已获得自交繁殖十二年的高代自交系材料。相对于诱导创制初期，材料表现出的结实率低，同种系单株间的差异较大；高代材料已表现出较显著的结实率提升和较一致的农艺性状表型。 本实验选取课题组多年培育的同源四倍体水稻高代自交系材料，通过形态学、农艺性状和细胞遗传学比较，研究水稻同源四倍体与二倍体之间的异同。结果显示，所有同源四倍体材料的染色体组成均为2N=4X=48，花粉母细胞(PMC)减数分裂行为较正常，99%以上的染色体都能在减数分裂中期I（MI）发生联会、配对，形成四价体和二价体，这与理论染色体组构成相符。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。统计分析表明，二价体和四价体的比例对结实率没有显著影响，但是三价体的数目对结实率有一定影响。这一结果表明了结实率和细胞减数分裂行为可能存在相关性，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依据。 然后，对同源四倍体水稻高代自交系材料进行农艺性状和品质性状的统计与分析。主要针对结实率、每穗实粒数、有效分蘖和穗长等主要农艺性状，以及直链淀粉含量这一重要的品质性状进行统计。将统计结果与1996年诱导加倍的初代材料的数据相对比分析，结果显示所有材料经过多代选育培养，其农艺性状已经有了较显著的提高，同时同源四倍体材料的农艺性状稳定性也有了较显著的提升。如结实率的提高幅度较大，所有材料的平均结实率均显著高于加倍初代，而同种材料不同单株间的结实率差异也显著地减少，变异系数（CV）的平均值由1996年的41.15%减少到了2008年的28.81%。其他重要农艺性状也有不同程度的改良，种内变异系数也相应地降低。此外，实验测量了同源四倍体材料和来源二倍体材料的直链淀粉含量。分析结果显示，部分材料的直链淀粉含量与二倍体亲本产生了较显著的差异，这可能是诱导加倍过程中的遗传变异造成的；同源四倍体材料的种内变异系数（CV）平均值由1996年的6%下降到了2008年的3.88%，显示出在品质性状方面，同源四倍体材料的遗传稳定性也有较显著的增加。同源四倍体材料农艺性状经过多年的选育，表现出一定的提升，同时，经过多年自交纯化，所有材料种系内的性状差异逐渐缩小，说明同源四倍体水稻的遗传稳定性随着自交纯化而增强，这为同源四倍体水稻的进一步选育打下了良好的基础。 最后，通过测量连续两年的自交系材料的遗传多态性，分析材料间遗传差异和种群遗传结构，深入研究连续两代材料间的遗传差异，研究同源四倍体水稻与二倍体材料遗传稳定性之间的差异。实验采用18对SSR微卫星标记对连续两代15个材料，共94份样本进行差异分析。通过扩增条带长度多态性分析，计算不同材料以及同种材料不同世代间的遗传距离，构建同源四倍体和二倍体水稻的分子指纹库，并绘制聚类图。结果显示，同源四倍体和二倍体不同材料间的遗传差异比较大，遗传距离处于0.4757至0.2816之间；而相同品种不同世代材料间的遗传差异较小，但也表现出一定的遗传差异。同种同源四倍体材料不同世代间的遗传差异比二倍体材料更大，两代四倍体材料间遗传距离处于0.1359至0.0485之间；而两代二倍体材料间的遗传距离处于均小于0.0388。结果表明，同源四倍体水稻高代材料具有一定的遗传稳定性，但与来源二倍体材料相比，其世代间的遗传变异性仍然较强。这种结果说明，经过多代的自交纯化培育，同源四倍体水稻材料能够建立起相对稳定的遗传结构，同时，其强于二倍体亲本的变异性有能够为新品种的选育，农艺性状、品质性状的改良提供一定的遗传基础。此外，分析结果表明通过分子标记辅助检验，水稻材料间的遗传多态性能够有效地区分不同的品种，这为水稻品种的分子鉴定提供了一定的依据。 本研究从细胞学鉴定，农艺性状统计分析以及分子标记辅助聚类分析多方面地对同源四倍体水稻高代系进行了研究，对探究同源四倍体水稻的遗传规律，进一步揭示其遗传特性、农艺性状的遗传构成，为进一步选育优质的多倍体水稻提供了一定的理论依据。 This group insists on creating new Autotetraploid Rice (Oryza sativa L.) materials, while improving the seed-setting of them for many years, cultivated and selected the inbred line materials, has obtained the high generation inbred lines after twelve years cultivation. Compared to the early induced materials, which shown the low seed setting, and the large difference between the different plants in the same germ-line; the high generation materials have shown significant improvement in seed setting and more uniform phenotype agronomic traits. The autotetraploid rice high generation inbred lines material, which has been cultivated for more than 12 years, was chose in this experiment. The similarities and differences between autotetraploid and diploid rice was studied through morphological, agronomic and cytogenetic ways. The results showed that all the chromosome of autotetraploid materials are composed of 2N=4X=48, the pollen mother cells (PMC) meiosis behavior is normal, more than 99% chromosomes in metaphase I(MI) were federated and paired to form tetravalents or bivalents, which constitutes a consistent theory of genome. In the meiosis process, the material with a higher seed setting showed less chromosome abnormal than the material whose seed setting is lower. However, statistical analysis showed that the bivalent and tetravalent rate had no significant impact on seed setting, but the number of trivalent had a certain impact on seed setting. The result shows that the seed setting may be related to the meiosis behavior, which provides a basis to cultivate new autotetraploid germ line with high seed setting through the meiotic behavior. Furthermore, the agronomic and quality traits of autotetraploid rice high generation inbred material were statistically analyzed. The statistically analysis was focused on major agronomic traits such as: seed setting, grains per panicle, effective tillers and panicle length, as well as the important quality trait amylose content. The statistic data was compared with the data in 1996, when the first induced generation of autotetraploid material, and the result shows that after a multi-generation breeding, the agronomic traits has been significantly improved in all the materials, while the stability of agronomic traits also significant upgraded. For instant, the seed setting increased significantly, the average seed setting of all materials was significantly higher than the first induced generation, and the differences between different plants in the same species also significantly reduced, the average of the coefficient of variation (CV) was reduced from 41.15% in 1996 to 28.81% in 2008. Other important agronomic traits had improved in different degrees; the coefficient of variation within species is also reduced accordingly. In addition, the amylose content of autotetraploid and diploid materials was measured in this experiment. The results shows that the amylose content of some of the material differed from diploid parents significantly, it may caused by the genetic change during the inducing, autotetraploid materials intra-specific coefficient of variation (CV) average reduced from 6% in 1996 to 3.88% in 2008, shows that this is a significant increase of quality traits stability in autotetraploid rice. Agronomic traits of autotetraploid material shows some improvement after years of breeding, at the same time, after years of purification, all material within the germ-line gradually narrow the differences in traits indicates that autotetraploid rice genetic stability was enhanced, which laid a good foundation for the further autotetraploid rice breeding. Finally, this experiment studied the genetic differences between materials of two generations and researched the difference of genetic stability between diploid and autotetraploid rice materials through investigating the genetic polymorphism, genetic differences between materials and population genetic structure of inbred line materials of two consecutive years.18 pairs of SSR microsatellite markers for 15 materials of two generations were used in this experiment, and the total of 94 samples were analyzed. Through the amplification length polymorphism analysis of different materials and materials in different generations, the genetic distance between materials and generations was analyzed, a diploid and autotetraploid rice molecular fingerprint database and map rendering cluster were constructed. The result shows that the genetic distance is between 0.4757 to 0.2816 among different autotetraploid and diploid materials; the genetic distance between different generations of same species was less, but also shows a certain degree of genetic differences. The inter-generational genetic differences of autotetraploid materials were greater than of the diploid materials, which are 0.1359 to 0.0485 as the genetic distance; comparing with the 0.0388 of diploid materials. The result shows that high generation inbred autotetraploid rice material has a certain genetic stability, but the genetic variation between generations is still strong comparing with the source diploid materials. It indicates that, after many generations of purification cultivation, autotetraploid rice materials established a relatively stable genetic structure, at the same time, stronger variability than its diploid parents are useful in the breeding of new varieties, provides a genetic foundation to the agronomic and quality traits improvement. In addition, the analysis result shows that the through the molecular marker-assisted testing, rice genetic polymorphism between materials can effectively distinguish different species, provides a certain basis for molecular identification of varieties of rice. A series of investigation such as cytological identification, statistical analysis of agronomic traits, molecular marker-assisted cluster analysis was applied in this experiment to research genetic pattern of autotetraploid rice high generation inbred lines, revealed the genetic characteristics and the genetic composition of agronomic traits, provides a theoretical basis for the further selection of high quality autotetraploid rice.

密度对三种豆科牧草生产力和水分利用率的影响

研究播种密度对3种豆科牧草生产力和水分利用效率的影响,以探讨其在草地农业系统中的合理优化模式。结果表明:沙打旺(Astragelus adsurgens)在高密区第2年生产力最高(18321 kg.hm-2),第3年开始下滑;苜蓿(Medicago sativa)在中、高密区生产力的增幅和增量差别不显著,以第3年最高(22563和22108kg.hm-2);胡枝子(Lespedeza daurica)在中密区第3年最高(7856 kg.hm-2);加大密度使沙打旺提早进入生长盛期,第2年即达到高生产力和较高水分利用效率;苜蓿以中密区(20~30株/m2)效果最好;达乌里胡枝子的生产力和水分利用率偏低,在黄土塬区草地农业中没有优势。

不同轮作施肥对土壤有机氮分解特征的影响

在各长期轮作施肥系统中,耕层土壤矿化培养过程的pH及剖面矿化势的变化,说明了长期不同的轮作和施肥方式引起土壤有机氮数量和性质的变化。pH变化具有一共同特征,在0~4 周,各处理间差异显著;而后期变化趋势趋于一致,向微碱性环境发展。各处理的剖面矿化势的变化反映了有机氮可矿化数量的改变。小麦连作施肥和苜蓿连作施肥均说明施用有机肥对土壤有机氮水平有较好的提高作用。粮豆轮作3 a对土壤供氮能力的提高作用大于粮饲轮作4 a和裸地处理。

黄土塬区3种豆科牧草对土壤水分的消耗利用研究

在田间完全旱作条件下采用3个密度和2种播种方式观察了3种多年生豆科牧草生长第2年对土壤水分的消耗利用情况。结果表明:苜蓿主要耗水深度在2~3 m,最深可达5 m,其中、高密度处理3 m以上土壤水分含量都在稳定田间持水量之下,已经开始形成土壤下伏干层;沙打旺耗水深度在0~2 m,最低含水量(11.61%)处于80~100 cm,在雨季可以恢复到稳定田间持水量之上;达乌里胡枝子主要耗水深度在1 m以上,最低含水量也在稳定田间持水量之上。单播沙打旺、苜蓿和达乌里胡枝子全生长期内对土壤水分的消耗分别为249.9、180.2和136.6 mm,水分利用效率分别是29.39、26.04和8.91 kg.mm-1.hm-2。混播、加大播种密度都会增加3种牧草土壤水分消耗,降低土壤储水量,提高干草产量和水分利用效率,但影响程度因牧草种类、播种方式以及不同的生长时段而异。

苜蓿干草和秸秆组合体外发酵营养特性及其利用研究

应用体外发酵产气技术,评价了苜蓿Medicago sativa干草和玉米Zea mays、小麦Triticum aestivum秸秆分别以0∶100、25∶75、50∶50、75∶25和100∶0进行两两组合时的发酵特性。结果表明：不同比例组合产气量（GP）、理论最大产气量（A）、产气速率（b）及产气延滞时间（LAG）变化趋势不同;苜蓿干草与玉米秸秆按50∶50的比例或苜蓿干草与小麦秸秆按75∶25的比例组合时的效应明显好于其他组合。48 h产气量与粗蛋白（CP）（P〈0.05）及中性洗涤可溶物（NDS）含量呈正相关关系,而与中性洗涤纤维（NDF）、酸性洗涤纤维（ADF）、半纤维（HC）含量及NDS/CP（P〈0.01）呈负相关关系;理论最大产气量与CP、NDS的含量呈正相关关系,与NDF、ADF、HC和NDS/CP（P〈0.01）呈负相关关系;产气速率与CP（P〈0.01）、HC（P〈0.01）、NDS（P〈0.01）呈极显著正相关关系,分别与NDF（P〈0.01）、ADF（P〈0.01）、NDS/CP（P〈0.05）呈负相关关系;产气延滞时间与饲草料的主要营养成分的相关关系不明显,只与NDS/CP（P〈0.05）呈显著正相关关系。结论认为,饲草中非结构性碳水化合物与蛋白质比例决定了体外发酵产气的特性。生产实践中应针对低质粗饲料营养特性,适当添补易发酵或高蛋白牧草,提高粗饲料利用效率。

青海东部4种作物群落蒸散规律研究

采用改进的离体连续快速称重法对7~9月燕麦(Avena Sativa)、豌豆(Pisum Sativum)、无芒雀麦(Bromus Innermis)、紫花苜蓿(Medicago Sativa)枝和叶片的蒸腾速率、蒸腾量及土面蒸发量进行了测定。结果表明,豌豆的枝蒸腾速率和叶片蒸腾速率均最高,其次为紫花苜蓿和燕麦,无芒雀麦最低。在7~9月中,8月4种作物整株和叶片的平均日蒸腾量均最高,9月次之,7月最低。4种作物群落土面蒸发量的日变化无明显规律性。

The Genomes of Oryza sativa: A history of duplications

We report improved whole-genome shotgun sequences for the genomes of indica and japonica rice, both with multimegabase contiguity, or almost 1,000-fold improvement over the drafts of 2002. Tested against a nonredundant collection of 19,079 full-length cDNAs, 97.7% of the genes are aligned, without fragmentation, to the mapped superscaffolds of one or the other genome. We introduce a gene identification procedure for plants that does not rely on similarity to known genes to remove erroneous predictions resulting from transposable elements. Using the available EST data to adjust for residual errors in the predictions, the estimated gene count is at least 38,000 - 40,000. Only 2% - 3% of the genes are unique to any one subspecies, comparable to the amount of sequence that might still be missing. Despite this lack of variation in gene content, there is enormous variation in the intergenic regions. At least a quarter of the two sequences could not be aligned, and where they could be aligned, single nucleotide polymorphism ( SNP) rates varied from as little as 3.0 SNP/kb in the coding regions to 27.6 SNP/kb in the transposable elements. A more inclusive new approach for analyzing duplication history is introduced here. It reveals an ancient whole-genome duplication, a recent segmental duplication on Chromosomes 11 and 12, and massive ongoing individual gene duplications. We find 18 distinct pairs of duplicated segments that cover 65.7% of the genome; 17 of these pairs date back to a common time before the divergence of the grasses. More important, ongoing individual gene duplications provide a never-ending source of raw material for gene genesis and are major contributors to the differences between members of the grass family.