同源四倍体与二倍体水稻的遗传差异及产量、品质性状研究

本文以中国科学院成都生物研究所培育的同源四倍体水稻和二倍体水稻为材料，进行遗传差异及产量、品质性状的研究：1、以二倍体水稻为对照，研究了同源四倍体水稻在2004 年和2005 年的结实情况。结果同源四倍体的花粉育性、结实率均不同程度下降，尤其低代材料更是大幅度下降。F 检验表明，同源四倍体不同个体间的各产量性状均差异显著，说明其具有很大的遗传改良潜力。从1996 年到2005 年对部分同源四倍体水稻进行了连续选择改良，T 检验表明经过9 年的选择改良，其结实率显著提高。本文还对同源四倍体水稻各产量性状间的相关性进行了分析,结果结实率与花粉育性、穗着粒数、穗实粒数极显著相关；理论产量与花粉育性、有效穗数、穗着粒数、穗实粒数、结实率及千粒重极显著相关。i2、用(CT)n 微卫星标记和PCR-Acc Ι分子标记对40 份同源四倍体和14 份二倍体水稻Wx 基因进行研究。结果，(CT)n 微卫星标记检测，Wx 基因呈Wx1、Wx2 和Wx3 3 种多态性；PCR-Acc Ι 检测，Wx 基因表现为G-型和T-型。测定稻米直链淀粉含量(AC)、胶稠度(GC)和糊化温度(GT)，并探讨其与Wx 基因的关系，结果，二倍体和同源四倍体水稻均存在：Wx 基因型相同，AC 差异较小，Wx 基因型不同时，AC 差异较大，Wx1 基因型品种AC 最高， Wx2 基因型品种AC 次之，Wx3 基因型品种AC 最低；基因型相同时，同源四倍体AC 低于二倍体；同源四倍体与对应二倍体间，Wx 基因型相同时，AC 差异很小；而Wx 基因型发生变异时，AC 差异很大。同时，进行相关性分析，结果二倍体和四倍体水稻均存在AC、GC 与Wx 基因密切相关；而GT 与Wx 基因相关不显著。综合分析，(CT)n微卫星标记与PCR-Acc Ι 分子标记检测的相关系数为0.842，呈极显著正相关，可以将其结合起来进行同源四倍体新品种的选育和改良。3、利用RAPD 技术，对同源四倍体和二倍体水稻的遗传差异进行分析。17条引物在同源四倍体中扩增出178 条带（PPB＝81.5%），在二倍体中扩增出173条带（PPB＝76.3%）；同源四倍体和二倍体的Shannon 遗传多样性指数分别为0.4848 和0.4679，多态信息量分别为0.3301 和0.3216，遗传距离分别为0.3572和0.3460；同源四倍体与其对应二倍体间遗传距离为0.1914-0.4633，平均为0.2914。表明同源四倍体的遗传多样性高于二倍体，且同源四倍体与其二倍体之间存在较大的遗传差异，这些将为水稻品种改良和新品种选育提供科学依据。上述产量、品质性状及遗传差异分析的结果，不仅有利于加快同源四倍体水稻的遗传改良进程，而且为进一步研究、利用同源四倍体水稻奠定了初步基础。 AbstractStudy on genetic diversity, yield characters and quality traitsof autotetraploid and diploid riceLiu Yuhua (Botany)Directed by Associate Prof. Tu ShengbinIn this study, diploid and autotetraploid rice, which were cultivated in ChengduInstitute of Biology were analyzed in genetic diversity, yield characters and qualitytraits.In the study, 2 diploid and 29 autotetraploid rice(2n=4x=48) materials, including4 preliminary and 25 advanced generation, were investigated for yield characters.Compared with diploid check, the pollen fertility and seed set of autotetraploiddeclined dramatically, especially in preliminary generation. F-test indicated that therewere remarkable differences among different varieties, showing that autotetraploidmaterials had strong potential for improvement. From 1996 to 2005, someautotetraploid rice had been selected and improved. T-test showed that seed setincreased obviously. The relationships among yield characters of autotetraploid ricewere analyzed. Seed set was strongly correlated with pollen fertility, total grainnumber per panicle and productive grain number per panicle; theoretical yield wasstrongly correlated with pollen fertility, productive panicle number per plant, totalgrain number per panicle, productive grain number, seed set and 1000-grain weight.Wx genotypes of 40 autotetraploid rice and 14 diploid rice were tested by usingthe (CT)n microsatellite marker and a cleaved amplified polymorphic sequence(CAPS) molecular marker named PCR-Acc Ι. Three microsatellite alleles wereproduced, i.e. Wx1, Wx2 and Wx3 both in autotetraploid and in diploid rice.Comparatively, PCR- Acc Ι molecular marker produced two genotypes, G-type andT-type for both autotetraploid and diploid rice. In this study, amylose content (AC), gel of consistency (GC) and gelatinization temperature (GT) of rice grain weremeasured and their relationships with Wx alleles were analyzed. The results showedthat variation of AC between autotetraploid and diploid rice was small when they hadthe same Wx genotype. However, variation of AC turned to be large when the Wxgenotypes were different. Actually, AC met the maximum value in Wx1 varieties andWx2 varieties the middle and Wx3 varieties the minimum. And AC was lower inautotetraploid than in diploid. Correlation analysis was done in this experiment. ACand GC of rice grain were probably controlled by Wx gene or other gene whoselocation was strictly linked to Wx gene, while GT of rice was not. The correlationcoefficient between Wx genotypes which revealed by (CT)n microsatellite marker andPCR-Acc Ι molecular marker was 0.842 with significant level. That revealed aconsistent result between the two types of markers. So it was possible to utilize boththe two types of markers to select and promote germplasm of autotetraploid rice.RAPD molecular markers were used to analyze the genetic diversity betweendiploid and autotetraploid rice. 178 repeatable bands were detected through 17 RAPDprimers with percentage of polymorphic bands was 81.5% in autotetraploid rice while173 repeatable bands were detected with percentage of polymorphic bands was 76.3%in diploid rice. According to the measurement of Shannon index, polymorphicinformation content and genetic distance, genetic diversity of autotetraploid was on ahigher level, genetic variation between autotetraploid and diploid rice was relativelyhigh. All these contributed to the genetic selection and improvement in rice breeding.As mentioned above, the results are not only helpful to promote the process ofrice improvement, but also to confirm the basic for further study of autotetraploid rice.

ACHT操作引起黑麦(Secale cereale)染色体的畸变和重建

用ACHT处理黑麦萌动种子，对修复前后材料的观察和分析结果表明：1. ACHT操作引起染色体数目变化和染色体断裂损失。在一定 条件和范围内，不同处理引起的这种变化具有显著差异，条件越剧烈，染色体数目变化的范围和频率愈大，断片发生的数量和频率 也愈高，同时修复前后染色体数目的变化范围和频率与断片发生的数量和频率以及它们的修复频率均表现明显的相关性。2. ACHT 操作引起染色体畸变的多样性。经ACHT处理后，胚根细胞染色体有4种断裂方式，包括着丝粒断裂、次溢痕断裂、长臂断裂和短臂 断裂等，其中着丝断裂频率最高；产生6种断片类型，包括有着丝粒和端粒的、有着丝粒而无端粒的、有部分着丝粒和端粒的、有 部分着丝粒而无端粒的、只有端粒的、既无着丝粒也无端粒的断片等。3. ACHT操作引起遗传结构重建的多样性。经ACHT处理后， 对修复24-72小时材料进行核型比较(按Stebbins 和 Levan 标准)和随体分析，处理细胞在染色体数目、大小、形态、位置等方面 均发生显著变化，说明ACHT处理使这些细胞的染色体结构和染色体组型发生了深刻变化。进一步通过Giemsa C— 带分析，观察到 多种重建染色体类型，包括易位型染色体、附加型染色体、无着丝粒染色体、化染色体、增加的m染色体以及某些带型特异的染色 体等。4. RAPD 分析从分子水平上验证了ACHT能有效地引起遗传结构的改变。所用10种引物对处理和对照材料基因组DNA的扩增产 物在条带数目、条带位置及带型特征等方面均有明显差异，其中4种引物出现条带减少，6种引物出现条带增加，后者还包括一个带 位移动。这说明两种材料的基因组DNA具有明显的RAPD反应多态性差异。This paper descripes some results draw on the basis of the observation and analysis on the rye before and after repaired through treating its budding seeds by ACHT in contrast to without ACHT: 1. ACHT manipulation caused the number variation and breakage damage of rye chromosome. Within certain conditions and timits, this phenomenon caused by different treats had signifcant difference: the more the treatment condition is drastie, the more the chageable range and frequence of rye chromosomae number, and so is the produced fragments. Meanwhile, there existed striking relationship among the changeable range and frequence of rye chromosome, the produced number and frequence of fragments and repairing frequence. 2. ACHT manipulafion engendered the diversify of rye chromosomal aberration. Four breakage patterns and six sorts of fragment were observed by watching the chromosome of the rye radicle treated by ACHT, including centric breakage (occuring in the highest frequence), secondary constriction breakage, long arm breakage and short arm breakage to the former, Comprising that with both centromere and telomere, that with centromere and without telomere, that with partial centromere and with telomere, that with partfial ceetromere and without telomere, that only with telomere and that neither with centromere nor with telomere, etc. 3. ACHT manipulation engendered the diversify or rye genetic structs reconstruction. Karureotype analysis(according to Stebbins and Levan) and satellite anaeysis were carried out to rye radicle through 24-72-hour-long recoverage after ACHT manipulation, which showed remarkable change happened on the rye chromosomal number、shape、arm ration and pattern, etc. and also on the satellite number、size、shape and location etc. Those indicated that ACHT manipulation engendered violent changes to rye chromatin structure and chromosome type. Further Giemsa C-banding analysis showed many types of reconstructed chromosome, such as translocation、addition、without centromere、st and other chromosome. 4. RAPD analysis checked the validity of ACHT on changing genetic structure of rye on the level of molecular biology. The treated and recovered rye has different amplifying band pattern by using IO valid arbitary primers selected from 40 comparing with the control.

碳离子辐射大丽花矮化突变体的RAPD分析

大丽花经兰州重离子加速器提供的80MeV/u12C6+离子束辐照后产生矮化突变体,用随机扩增多态性(Random Amplified Polymorphic,RAPD)DNA技术对野生型和突变体进行检测分析。结果表明,在所用的25条引物中,1.80×108/cm2剂量辐照后有18条引物扩增出现多态性片断,扩增条带多态率19.57%;1.08×108/cm2剂量辐照后仅有6条引物扩增出现多态性片断,扩增条带多态率5.76%。用Jaccard公式对扩增产物进行统计分析,结果表明,两种剂量C6+辐照后与对照相似性系数分别为0.65和0.92。高剂量辐射后DNA易发生突变,在品种改良和诱变育种中相对较高剂量的选择可能更为有效。

重离子束辐照对苜蓿外植体离体培养的影响及下胚轴再生体的RAPD分析

采用初始能量为100MeV/u的重离子束12C6+对紫花苜蓿下胚轴及子叶外植体进行辐照处理,研究重离子辐照对愈伤组织诱导状态及诱导率、愈伤组织相对生长率、体细胞胚诱导率及植株再生的影响。结果表明,重离子辐照对下胚轴及子叶愈伤组织的诱导具有抑制作用,且出愈率随着辐照剂量的增大而降低;在继代培养过程中,其愈伤组织相对生长率均高于对照组,外植体本身对重离子辐照所造成的损伤具有恢复能力;辐照处理对体细胞胚胎诱导也有影响,30Gy时,下胚轴诱导的体细胞胚胎发生较对照组早,数量多,较早地得到再生植株;而50Gy时,所得到的体细胞胚未能发育成再生植株。同时应用随机扩增多态性DNA(Random amplified polymorphic DNA,RAPD)技术对重离子辐照处理下胚轴所得再生植株进行检测分析,结果表明:所采用的20条随机引物中有11条在对照及处理组所得再生植株之间扩增出差异性多态条带,表明了重离子辐照引起苜蓿再生植株基因组DNA发生变异。

~(12)C~(6+)重离子辐照胡麻种子初步研究

本文研究了6×108cm-2、1.8×109cm-2和3.6×109cm-2的12C6+重离子束辐照对胡麻种子M1代生物学性状和DNA分子多态性等方面的影响。6×108cm-2辐照处理可引起胡麻发芽率提高,促进植株株高,增强花粉活力。同时辐照处理使胡麻种子千粒重和含油量有不同程度提高,辐射剂量越高,两者数值越大,3.6×109cm-2辐射剂量的胡麻种子千粒重和含油量与对照组的相比分别高出了16.5%和19.9%,此外在此剂量处发现了花粉发生了形态变化。辐照处理对胡麻DNA分子也产生了影响,筛选出的14个随机引物可以扩增出清晰、稳定、重复性好的DNA片段,有52个是多态性DNA片段,比率为52.5%。

用随机扩增多态性DNA技术对重离子辐照大丽花花色突变体的初步研究

品系为“大雪青”的大丽花幼枝,经兰州重离子加速器提供的80MeV/u12C6+束辐照后,种植在甘肃省定西市临洮新兴花卉公司基地。辐照8Gy的幼枝有一株花色突变体,用随机扩增多态性DNA(Random amplified polymorphic DNA,RAPD)技术对突变体和野生型进行检测分析。琼脂糖凝胶电泳结果表明,所用的10条引物中,有7条引物共扩增出78条带,其中5条引物扩增出了11条多态性片段,从而在DNA水平上证实了两者之间存在着差异,为进一步探讨重离子诱变机理打下基础。

空间搭载水稻种子后代基因组多态性及其与空间重离子辐射关系的探讨

神舟三号飞船搭载带核径迹辐射探测器的水稻种子装置,回收后应用随机扩增多态性DNA(randomamplified polymorphic DNA,RAPD)技术,分析了201粒升空种子长出植株的基因组多态性。在检测的189个基因座位范围内,30.2%的植株中发现与地面对照不同的扩增带,单株的多态性座位数为1￣25。特异扩增带的测序及单核苷酸多态性(single-nucleotide polymorphism,SNP)分析进一步证明了空间搭载水稻种子确实可导致当代植株基因组发生变异。同一技术分析个别种子连续世代的基因组多态性,结果显示,当代的部分多态性可遗传至后代。7粒受空间高原子序数、高能粒子轰击的种子,在当代植株均显示不同程度的基因组多态性,从胚受粒子击中的3粒种子后代中,筛选出农艺性状明显变异的突变株系,初步暗示了空间高能重离子辐射对诱导基因组的多态性,乃至遗传性表型变异的有效性。

Flower color chimera and abnormal leaf mutants induced by C-12(6+) heavy ions in Salvia splendens Ker-Gawl.

The effect of C-12(6+) heavy ions bombardment on mutagenesis in Salvia splendens Ker-Gawl. was studied. Dose-response studies indicated that there was a peak of malformation frequency of S. splendens at 200 Gy. Abnormal leaf mutants of the bileaf, trileaf and tetraleaf conglutination were selected. Meanwhile, a bicolor flower chimera with dark red and fresh red flower was isolated in M1 generation of S. splendens. Random amplified polymorphic DNA (RAPD) analysis demonstrated that DNA variations existed among the wild-type, fresh and dark red flower shoots of the chimera. The dark red flower shoots of the chimera were conserved and cultivated at a large-scale through micropropagation. MS supplemented with 2.0 mg/L BA and 0.3 mg/L NAA was the optimal medium in which the maximum proliferation ratio (5.2-fold) and rooting rate (88%) were achieved after 6 weeks. Our findings provide an important method to improve the ornamental quality of S. splendens.

Study on DNA Damage Induced by Neon Beam Irradiation in Saccharomyces Cerevisiae

Yeast strain Saccharornyces cerevisiae was irradiated with different doses of 85 MeV/u Ne-20(10+) to investigate DNA damage induced by heavy ion beam in eukaryotic microorganism. The survival rate, DNA double strand breaks (DSBs) and DNA polymorphic were tested after irradiation. The results showed that there were substantial differences in DNA between the control and irradiated samples. At the dose of 40 Cy, the yeast cell survival rate approached 50%, DNA double-strand breaks were barely detectable, and significant DNA polymorphism was observed. The alcohol dehydrogenase II gene was amplified and sequenced. It was observed that base changes in the mutant were mainly transversions of T-->G and T-->C. It can be concluded that heavy ion beam irradiation can lead to change in single gene and may be an effective way to induce mutation.

Nucleation and growth of glycine crystals with controllable sizes and polymorphs on Langmuir-Blodgett films

Control of crystal polymorph and size is very important in many application fields. Herein we demonstrate that Langmuir-Blodgett (LB) films of stearic acid (SA) and octadecylamine (ODA) can serve as templates and generate different polymorphs of glycine crystals. In the neutral aqueous solutions, gamma-glycine crystallizes on LB films of ODA while the polymorphic outcome becomes the (x-form on LB films of SA. These observed results could be explained by the electrostatic interactions and geometric lattice matching at the LB film/crystal interfaces, respectively. By keeping the appropriate supersaturation, we have successfully controlled the number of crystals grown on LB films; for example, in some certain cases, only one piece of crystal was grown on LB films in solution. Therefore, large crystals of centimeter size could be prepared. These experimental results suggest a new approach to produce an organic crystal with bulk scale.

Supramolecular isomerism in zinc hydroxide coordination polymers with pyridine-2,4-dicarboxylic acid: Two polymorphs with centrosymmetric two-dimensional and acentric three-dimensional coordination networks

Reactions of Zn(BF4)(2) and pyridine-2,4-dicarboxylic acid (2,4-pydcH(2)) in the presence of 1,2-bis( 4-pyridyl) ethylene or 1,3-bis(4-pyridyl) propane under hydro(solvo) thermal conditions yielded two polymorphic metal-organic coordination polymers formulated as Zn-2(OH)(2)(2,4-pydc) (1 and 2). Polymorph 1 features a two-dimensional (2-D) layer-like structure that is constructed by 2,4-pydc ligands bridging between the Zn-OH-Zn double-chain units. Each single Zn-OH-Zn chain is composed of mu(2)-OH groups connecting trigonal bipyramidal and tetrahedral Zn centers. Polymorph 2 is a 3-D coordination polymer containing 2-D Zn-OH-Zn sheets that consist of mu(2)- and mu(3)-OH groups and trigonal bipyramidal Zn centers. The sheets are pillared by 2,4-pydc ligands to form an acentric structural architecture. 1 and 2 are rare examples that the two polymorphs exhibit a centrosymmetric 2-D coordination network and an acentric 3-D coordination network, respectively. The different structures lead to differences in photoluminescent properties and thermal stabilities for 1 and 2.

A new oxide ion conductor: La3GaMo2O12

A new oxide ion conductor, La3GaMo2O12, with a bulk conductivity of 2.7 X 10(-2) S.cm(-1) at 800 degrees C in air atmosphere was prepared by the traditional solid-state reaction. The room temperature X-ray diffraction data could be indexed on a monoclinic cell with lattice parameters of a=0.5602(2) nm, b=0.3224(1) nm, c= 1.5741(1) nm, beta= 102.555(0)degrees, V=0.2775(2) nm(3) and space group Pc(7). Ac impedance measurements in various atmospheres further support that it is an oxide ion conductor. This material was stable in various atmospheres with oxygen partial pressure P(O-2) ranging from 1.0 X 10(5) to 1.0 X 10(-7) Pa at 800 degrees C. A reversible polymorphic phase transition occurred at elevated temperatures as confirmed by the differential thermal analysis and dilatometric measurement.

Single crystal structure of form I syndiotactic poly(butene-1)

The structures of single crystals of syndiotactic poly(butene-1) in form I, produced by thin-film growth, are studied by transmission electron microscopy and electron diffraction. Bright-field electron microscopy observation shows that the single crystal exhibits a regular rectangular shape with the long axis along its crystallographic b-axis. Electron diffraction results indicate an isochiral C-centered packing of a-fold helical chains in an orthorhombic unit cell corresponding to the C222(1) space group, according to the model proposed in the literature. The differences with the polymorphic behavior of syndiotactic polypropylene concerning the formation and the stability of the isochiral mode of packing are outlined.

Effect of lanthanum ions on the lipid polymorphism of phosphatidylethanolamines

The influence of lanthanum ions on the polymorphic phase of egg phosphatidylethanolamine and dielaidoylphosphatidylethanolamine (PE and DEPE) has been investigated by means of P-31-nuclear magnetic resonance (P-31-NMR) and high sensitivity differential scanning calorimetry (DSC) techniques. P-31-NMR experiments show that lanthanum ions promote the formation of the hexagonal II phase at temperatures lower than those of the pure egg PE, DSC results also show that lanthanum ions induce the formation of hexagonal II phase in DEPE liposomes even al very low ion concentration, The effect of lanthanum ions on the polymorphism of PE liposomes is much greater than that of calcium.

Growth rates of different polymorphs from interspherulitic boundary profiles

Quantitative data on the crystallization kinetics of polymorphic polymers can be derived from the investigation of gross spherulitic morphology formed in isothermal conditions. Depending on distance between centers, and the time lag between their formation and relative growth rates, various types of boundary lines can be generated by the impinging of two spherical bodies whose radii increase linearly with time, In polymorphic polymers, different types of spherulites often develop simultaneously at different rates from sporadic or predetermined nuclei. In same cases, the so-called growth transformation, in which a nucleus of the fast growing specie is formed at the tip of an advancing lamella of the slower crystal form, provides an alternative mode of nucleation, It is shown that if only one event of growth transformation takes place at the front of a slow growing body, the fast growing spherulite swallows the parent one and the resultant shape of interspherulitic boundary is described by two symmetrical logarithmic spirals whose parameters can be extracted from micrographs taken at the end of crystallization. These concepts are applied to determine the radial growth rate of gamma form spherulites of polypivalolactone in a wide range of temperatures through analysis of the alpha/gamma interspherulitic profiles formed in isothermal conditions and direct measurement of the growth rate of the alpha counterparts at the same temperature.