中国沙棘(Hippophae rhamnoides subsp. sinensis)天然群体的遗传变异分析

近十年，植物群体遗传学的研究飞速发展，然而与海拔相关的植物群体遗传结构和遗传变异研究却相对较少。到目前为止，还不清楚遗传变异与海拔之间是否有一个通用的格局。在山区，各种生态因子，如温度、降水、降雪、紫外线辐射强度以及土壤成分都随海拔梯度急剧变化，造成了即使在一个小的空间区域，植被类型变化显著，这种高山环境的异质性和复杂性为我们研究植物群体遗传结构和分化提供了方便。沙棘(Hippophea)属于胡颓子科(Elaeagnaceae)为多年生落叶灌木或乔木，雌雄异株，天然种群分布极为广泛。中国沙棘(H. rhamnoides subsp. sinensis)是沙棘属植物中分布较广的一个亚种，种内形态变异非常丰富，加之其具有独特的繁育系统和广泛的生态地理分布，是研究沙棘属植物遗传变异和系统分化的理想材料。本文从1,800 m 到3,400 m 分5 个海拔梯度进行取样，用RAPD 和cpSSR 分子标记研究了卧龙自然保护区中国沙棘天然群体的遗传结构和遗传变异。5 个取样群体依次标记为A、B、C、D 和E，它们分别代表分布在海拔1,800，2,200，2,600，3,000 和3,400 m 的5 个天然群体。RAPD实验用11 条寡核苷酸引物，扩增得到151 个重复性好的位点，其中143 个多态位点，多态率达94.7%。在5 个沙棘群体中，总遗传多样性值(HT)为0.289，B群体内的遗传多样性值为0.315，这完全符合沙棘这种多年生、远交的木本植物具有高遗传变异的特性。5 个群体内遗传多样性随海拔升高呈低－高－低变异趋势，在2,200 m海拔处的B群体遗传多样性达最大值0.315，3,400 m海拔处的E群体则表现最小仅0.098。5 个群体间的遗传分化值GST＝0.406，也即是说有40.6%的遗传变异存在于群体间，1,800 m海拔处的A群体与其它群体的明显分离是造成群体间遗传分化大的原因。UPGMA聚类图和PCoA散点图进一步确证了5 个群体间的关系和所有个体间的关系。最后，经过Mantel检测，遗传距离与海拔表现了明显的相关性(r = 0.646, P = 0.011)。cpSSR 实验中，经过对24 对cpSSR 通用引物筛选，11 对引物能扩增出特异性条带，只有2 对引物(ccmp2 和ARCP4)呈现多态性。4 个等位基因共组合出4 种单倍型，单倍型Ⅰ出现在A 群体的所有个体和B 群体的8 个个体中，C、D、E 三个群体均不含有，而单倍型Ⅱ出现在C、D、E 三个群体的所有个体及B 群体的18 个个体中，A 群体不含有。另外两种单倍型Ⅲ和Ⅳ为稀有类型，仅B 群体中的4 个个体拥有。这种单倍型分布模式和TFPGA 群体聚类图揭示了，C、D、E 群体可能来源于同一祖先种，而A 群体却是由另一祖先种发展起来的，B 群体则兼具了这两种起源种的信息，这可能是因为在历史上的某一时期，在中国沙棘群体高山分化的过程中，B 群体处某个或者某些个体发生了基因突变，具备了适应高海拔环境的能力，产生了高海拔沙棘群体的祖先种。 In recent ten years, studies about population genetics of plants developed rapidly,whereas their genetic structure and genetic variation along altitudinal gradients have beenstudied relatively little. So far, it is uncleared whether there is a common pattern betweengenetic variation and altitudinal gradients. In the mountain environments, importantecological factors, e.g., temperature, rainfall, snowfall, ultraviolet radiation and soil substratesetc., change rapidly with altitudes, which cause the vegetation distribution varying typically,even on a small spatial scale. The mountain environments, which are heterogeneous andcomplex, facilitate and offer a good opportunity to characterize population genetic structureand population differentiation.The species of the genus Hippophae L. (Elaeagnaceae) are perennial deciduous shrubs ortrees, which are dioecious, wind-pollinated pioneer plants. The natural genus has a widedistribution extending from Northern Europe through Central Europe and Central Asia toChina. According to the latest taxonomy, the genus Hippophae is divided into six species and12 subspecies. The subspecies H. rhamnoides ssp. sinensis shows significant morphologicalvariations, large geographic range and dominantly outcrossing mating system. Thesecharacteristics of the subspecies are favourable to elucidate genetic variation and systemevolution. To estimate genetic variation and genetic structure of H. rhamnoides ssp. sinensisat different altitudes, we surveyed five natural populations in the Wolong Natural Reserve at altitudes ranging from 1,800 to 3,400 m above sea level (a.s.l.) using random amplifiedpolymorphic DNA markers (RAPDs) and cpSSR molecular methods. The five populations A,B, C, D, and E correspond to the altitudes 1,800, 2,200, 2,600, 3,000 and 3,400 m,respectively.Based on 11 decamer primers, a total of 151 reproducible DNA loci were yielded, ofwhich 143 were polymorphic and the percentage of polymorphic loci equaled 94.7%. Amongthe five populations investigated, the total gene diversity (HT) and gene diversity within population B equaled 0.289 and 0.315, respectively, which are modest for a subspecies of H.rhamnoides, which is an outcrossing, long-lived, woody plant. The amount of geneticvariation within populations varied from 0.098 within population E (3,400 m a.s.l.) to 0.315within population B (2,200 m a.s.l.). The coefficient of gene differentiation (GST) amongpopulations equaled 0.406 and revealed that 40.6% of the genetic variance existed amongpopulations and 59.4% within populations. The population A (1,800 m a.s.l.) differed greatlyfrom the other four populations, which contributes to high genetic differentiation. A UPGMAcluster analysis and principal coordinate analyses based on Nei's genetic distances furthercorroborated the relationships among the five populations and all the sampling individuals,respectively. Mantel tests detected a significant correlation between genetic distances andaltitudinal gradients (r = 0.646, P = 0.011).Eleven of the original 24 cpSSR primer pairs tested produced good PCR products, onlytwo (ccmp2 and ARCP4) of which were polymorphic. Four total length variants (alleles) werecombined resulting in 4 haplotypes. The haplotype was present in all individuals of Ⅰpopulation A and 8 individuals of populations B, the other three populations (C, D and Epopulations) did not share. The haplotype was present in all individuals of populations C, D Ⅱand E and 18 individuals of populations B, population A did not share. The other twohaplotypes and were rare haplotypes, which were only shared in 4 individuals of Ⅲ Ⅳpopulation B. The distribution of haplotypes and TFPGA population clustering map showedthat the populations C, D and E might be origined from one ancestor seed and population Amight be from another, whereas population B owned information of the two ancestor seeds. Itwas because that gene mutation within some individual or seed in the location of population Bwas likely to happen in the history of H. rhamnoides, which was the original ancestor of thehigh-altitude populations.

不同小麦品种（系）中主要矿质元素的含量比较及与品质的关系

采用微波消解、电感耦合高频等离子体原子发射光谱（ICP-AES）的方法,对62份不同小麦品种（系）中锌、铁、铜、钙、钠和钾的含量进行了测定。同时利用红外线品质测定仪对主要品质指标粗蛋白、湿面筋、沉降值进行了测定。结果表明，不同小麦品种（系）中各种矿质元素的含量存在差异，2006年小麦品种中铁含量变幅为18.55-58.19 ug/g，平均为30.83ug/g ，最高与最低的相差39.64ug/g；锌含量变幅为5.70-25.80 ug/g，平均为15.13ug/g ，最高与最低相差20.10ug/g。2008年小麦品种（系）中铁含量变幅为16.68-52.25 ug/g，平均为30.10ug/g，最高与最低相差35.58ug/g；锌含量变幅为12.29-33.47 ug/g，平均为21.11ug/g，最高与最低相差21.18ug/g；钙含量变幅为167.53-348.80ug/g，平均为248.59ug/g，最高与最低相差192.59ug/g；铜含量变幅为2.32-5.83 ug/g，平均为2.98ug/g，最高与最低的相差3.61ug/g；钾含量变幅为1822.71-4414.91 ug/g，平均为2617.87ug/g，最高与最低的相差2634.72ug/g；钠含量变幅为10.25-39.82 ug/g，平均为23.05ug/g，最高与最低的相差29.57ug/g。 两年不同小麦品种（系）中矿质元素的含量分析结果表明：铁、铜、钙、钠和钾含量年际变化不明显,说明小麦对铁、铜、钙、钠和钾的吸收较稳定；锌含量变化较大，可能受环境的影响比较大。分析各矿质元素含量与粗蛋白、湿面筋、沉降值及元素之间的相关关系，结果表明，锌含量与粗蛋白含量呈极显著正相关关系，相关系数为0.317，与湿面筋含量之间呈显著正相关，相关系数达到0.246；铁含量与粗蛋白含量呈显著的正相关关系，相关系数是0.262；铜、钙、钠和钾含量与粗蛋白含量、湿面筋和沉降值之间存在正相关，但不显著，其中钠与沉降值之间为负相关。表明施锌或铁对提高小麦粗蛋白和湿面筋有显著效应，其余矿质元素有促进作用但不明显。 利用RAPD分子标记技术对川育23、41058、川育20及其父母本进行分析，力图从分子水平找到小麦矿质元素含量之间的差异性，琼脂糖电泳结果表明不同的小麦品种（系）间扩增出了差异条带。 以上研究结果，将对筛选“微量营养强化型”小麦新材料，选育“微量营养强化型”小麦新品种奠定基础。 62 different wheat cultivars was digested with HNO3 in a tightly closed vessel heated under micro-wave,then contents of zinc，iron,copper，calcium，sodium and potassium were determined by inductively coupled plasma-atomic emission spectroscopy(ICP-AES)．The main indexes of wheat quality such as total protein、wet glu and sedimentation volume were detected by Infratec 1255 Food & Feed Analyzer at the same time.The obtained results showed that variation for all of the mineral elements concentrations among different cultivars were observed .In 2006, the amplitude variation of the iron content was 18.55-58.19 ug/g,the average value was 30.83ug/g,and 39.64ug/g between the highest-content cultivar and the lowest one; the amplitude variation of the zinc content was 5.70-25.8 ug/g,the average value was 15.13ug/g,and 20.10ug/g between the highest-content cultivar and the lowest one.In 2008, the amplitude variation of the iron content was 16.68-52.25 ug/g,the average value was 30.10ug/g,and 35.58ug/g between the highest-content cultivar and the lowest one; the amplitude variation of the zinc content was 12.29-33.47 ug/g,the average value was 21.11ug/g,and 21.18ug/g between the highest-content cultivar and the lowest one; the amplitude variation of the calcium content was 167.53-348.80ug/g,the average value was 248.59ug/g,and 192.59ug/g between the highest-content cultivar and the lowest one; the amplitude variation of the copper content was 2.32-5.83 ug/g,the average value was 2.98ug/g,and 3.61ug/g between the highest-content cultivar and the lowest one; the amplitude variation of the potassium content was 1822.71-4414.91 ug/g,the average value was 2617.87ug/g,and 2634.72ug/g between the highest-content cultivar and the lowest one; the amplitude variation of the sodium content was 10.25-39.82 ug/g,the average value was 23.05ug/g,and 29.57ug/g between the highest-content cultivar and the lowest one. Analysis was made on the annual variation of mineral elements content in different Wheat cultivars ,the result shows:there is no obvious difference of iron ,copper ,sodium、calcium and potassium concentrations in wheat cultivars, suggesting the absorption of the iron, copper, sodium、calcium and potassium by wheat are relatively steady ,but zinc concentrations change obviously ,maybe influenced heavily by environment . The correlation between mineral elements 、mineral elements and total protein、mineral elements and sedimentation volume as well as mineral elements and wet glut were analysed in this paper, the result showed that there was significant positive correlation between zinc content and total protein (the correlation coefficient is 0.317), positive correlation between zinc content and wet glu (the correlation coefficient is 0.246), positive correlation between iron content and total protein (the correlation coefficient is 0.262). there was positive but not obvious correlation between the contents of copper, calcium, sodium or potassium and total protein, wet glut or sedimentation volume,among which was negative correlation between sodium and sedimentation volume.It was indicated zinc or iron fertilization has prominent effects in improving the total protein in wheat, the rest mineral elements have Non- obvious facilitation. The study then forecasted the genetic difference of different wheat by the molecular marker of RAPD in order to find differences in molecular level. Chuanyu23、41058、chuanyu20 as well as their male and female parents were analysed by RAPD markers,Agarose gel electrophoresis of DNA revealed the appearance of differential bands . The above-mentioned results of this study establish the foundation to screening the new materials of wheat of " strengthening type of micro- nutrition ", and to breeding the new wheat cultivars of" strengthening type of micro- nutrition ".

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.

小麦中微量元素锌铁硒的含量的分析

本研究应用微波消解ICP-AES 法对62 个小麦品种及3 个地区土壤的锌铁硒含量进行了分析测定，发现不同小麦品种中微量元素含量差异很大，姊妹系间也存在差异。含铁量最高与最低的小麦品种铁含量相差29.68mg/kg。含锌量最高与最低的小麦品种锌含量相差46.70 mg/kg。含硒量最高与最低的小麦品种硒含量相差0.056 mg/kg。对不同地点的小麦及土壤中锌铁硒含量进行方差分析，发现双流和西昌两地种植小麦的铁含量和硒含量均有显著差异，西昌和荣县种植的锌含量有显著差异。在3 个地点中双流种植小麦硒含量最高，西昌种植小麦的铁和锌含量最高。 通过对小麦微量元素含量与土壤中微量元素含量进行了相关性分析，结果表明：小麦中的锌铁含量与土壤中的锌铁含量呈显著正相关，土壤中铁与锌含量呈极显著正相关，小麦中铁与锌含量也呈极显著正相关。随着土壤微量元素锌铁的提高，小麦中的锌铁元素含量同时提高，而且小麦对两种元素的吸收互相促进。土壤中的硒含量与锌铁含量呈负相关。小麦中硒含量也与锌铁含量也呈负相关。说明锌和铁与硒互相拮抗。小麦硒含量与土壤硒含量呈正相关，但不显著。表明土壤硒含量可以影响小麦硒含量，但不是决定因素，小麦硒含量与小麦自身因素有关。 对姊妹系G290（高硒含量）和G289（低硒含量）进行抗重金属胁迫和抗旱性实验发现，高硒品种G290的抗逆性优于低硒品种G289。 利用RAPD 技术对7 个姊妹系进行遗传差异分析发现，高硒材料G290出现了特异条带，分别标为1、2、3、4，其他姊妹系品种中未发现特异条带，回收4 条特异条带并连接转化，得到目的片段1、2、3 的重组子，进行测序。NCBI 中结果显示没有找到植物中的同源序列，说明特异序列可能是未发现的基因片段，推测可能与小麦硒含量有关，有待进一步研究。 以上研究结果，对小麦营养研究及功能性小麦的筛选和栽培具有指导作用。 In this study, we determinated the contents of zinc, iron, selenium in 62 wheat cultivars and soil samples of three regions by method of microwave digestion/ ICPAES,found that there was great difference of zinc, iron, selenium contents in different wheat cultivars as well as different sister lines. Iron content difference was 29.68 mg/kg between the highest-iron-content cultivar and the lowest one, and zinc content difference was 46.70 mg/kg , selenium content difference was 0.056 mg/kg. Anova analysis was made on contents of zinc, iron, selenium in wheat and soil samples of different locations, significant differences of Fe and Se contents were found between wheat in Shuangliu and Xichang, significant difference of Zn content was found between wheat in Xichang and Rongxian. Se content in wheat of Shuangliu was highest, Fe and Zn contents in wheat of Xichang were highest. Relativity analysis was made on three trace elements in Wheat and in soil, the result showed that there was significant positive correlation of zinc, iron content between in Wheat and in soil, as well as between Fe and Zn both in wheat and in soil. With the improving of Zn, Fe contents in soil, contents of Zn and Fe in wheat increased and absorption of Zn and Fe in wheat will mutual promote. Negative correlation of Se and Zn contents was found in wheat and soil, but not significant, that meant the antagonism of Se and Zn. Positive correlation of Se content in wheat and soil was found. High selenium content G290 and low selenium content G289 in sister lines were selected for heavy metal stress and drought resistance experiments, the result showed that the resistance of high-selenium-content cultivar was better than low selenium one. Analysis on genetic difference was made by RAPD, and specific bands were selected, marked 1,2,3,4, no more specific bands were found in other sister lines.4 bands were recovered, ligated to T-vector and transformed E.coli. Three recombinant plasmids were obtained and sequenced. NCBI Blast showed there was no homology with other plants. It implied that these fragments probably be new genes and maybe were related to selenium in wheat. It needs further research. This paper would be useful for the study of wheat nutrition as well as selection and cultivation of functional wheat.

~(12)C~(6+)重离子辐照油菜诱变效应研究

用30Gy、90Gy和180Gy 12C6+重离子辐照油菜干种子,研究其对油菜M1代的诱变效应。结果表明不同剂量12C6+重离子辐照使油菜的出苗率、株高和开花率有不同程度的提高,并使开花期提早;30Gy辐照使单株角果数和单株产量有了一定程度的提高;三种辐照剂量都造成了花粉生活力、千粒重和含油量的降低。RAPD扩增结果表明,42个随机引物中有13个引物扩增出差异条带,30Gy、90Gy和180Gy引起的RAPD变异率分别为22.1%、23.7%和36.2%。研究表明,12C6+重离子辐射能有效地引起油菜DNA序列发生改变,从而诱导基因变异,为油菜育种提供丰富的种质材料。

~(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。

重离子辐照对红酵母的诱变作用

应用重离子加速器的 50MeV/u12C6+重离子对胡萝卜素生产菌-红酵母(Rhodotorula RYStrain )进行辐照处理,经酵母发酵实验,发现 50MeV/u12C6+重离子对胡萝卜素生产菌-红酵母具有诱变作用。初步筛选到了胡萝卜素产量有变化的辐照变异菌株,并对这些辐照变异菌株进行了 RFLP(限制性片段长度多态性)和 RAPD(随机扩增 DNA 多态性)分析,这些工作为工业上利用重离子对胡萝卜素生产菌进行诱变育种展现了新的前景。

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.

~(12)C~(6+)高能重离子辐照大葱损伤及其分子生物学效应；Cytological Damage and Molecular Biology Effect of ~(12)C~(6+) Heavy Ions on Allium fistulosum L.

利用30,90,180Gy3种剂量的12C6+重离子束辐照大葱种子,研究其在细胞水平和农艺性状的诱变效应并进行RAPD分析。通过与M1代的研究结果比较后表明:经过不同剂量12C6+重离子照射后能有效地诱导大葱细胞形成微核和染色体畸变,这种诱变效应,在M2代仍然有所表现。M1代大葱结果期的株高、白长、花序直径和种子产量随辐照剂量增加产生明显差别,其中30Gy辐照组增幅最大。大葱总水溶性蛋白质和维生素C的含量在30Gy组中积累最多,在90Gy组有明显下降。与M1代一致,M2代中大葱染色体微核率及RAPD分析所得的DNA多态性比率仍然与辐照剂量呈正相关,但比率整体下降;说明高能量重离子辐照造成的DNA变异在M2代被修复和淘汰。

重离子辐照甜高粱新品种选育及其诱变机理研究

利用重离子辐照技术对甜高粱种子进行不同剂量的诱变处理，并分析辐照后代的农艺性状、生理生化特性及基因组DNA的多态性差异，旨在选育出含糖量高、生物量高及抗逆性强的新品种，为发展生物质燃料乙醇产业提供优质的原料，并阐明重离子对甜高粱的诱变机理。主要结果如下： 1.甜高粱在田间的存活曲线表现为“类马鞍型”，随着辐照剂量的增加，其存活率先降后升再下降。 2.筛选出株高、单秆重、糖锤度、早熟型、茎粗等突变类型的材料，尤其是80Gy辐照剂量下从BJ0602中得到的早熟突变材料KFJT-1，生育期缩短了20天左右。 3.和未辐照株KFJT-CK相比，辐照突变株KFJT-1的萌芽指标表现为极显著差异（p＜0.01），其发芽势、发芽指数、活力指数和子叶长度、胚根鲜重及子叶鲜重分别下降了24％、12.69％、0.8108％和15.32％、76.27％、27.08％。 4. 利用RAPD技术对不同剂量的辐照处理检测出的多态性差异表明，不同剂量的碳离子束辐照后，不同辐照剂量对应的5种处理材料的DNA突变率分别0％、11.4％、12.2％、18.7％和17.7％。 重离子辐照可引起甜高粱各个方向的突变，有些突变材料生物量和含糖量均高，而有些突变材料表现出生长点消失、叶片扭曲、黄化等表型性状

重离子辐照普通小麦间接效应的研究

应用不同能量、不同剂量的碳离子和氧离子加照普通小麦，研究其M1代和M2代的生物学（形态）性状、染色体畸变率及微核率、生理生化和DNA分子等方面的变化。并对这些变化的生物学机理进行了探讨，结果如下：1.75MeV/u~(16)O~(8+)离子经过适当降能后注入小麦种子的不同部位和贯穿整个种子，对其M1代幼苗的可溶性蛋白质组分以及电脉中不同区段蛋白质组分相对含量加以分析，结果表明：1）同对照相比，随着辐照剂量的增加，所有辐照材料（包括贯穿和注入）第二区段（分子量较高区段）蛋白质组分的相对含量下降；而第五区段（分子量最小区段）蛋白质组分的相对含量升高。2）种子的贯穿处理，同时也引起第一区段（分子量最高区段）和第三区段蛋白质组分相对含量的下降以及第四区段的升高；其中第三、四区段的变化明显区别于注入效应。3）注入胚和胚乳的区别在于后者第一区段蛋白质组分相对含量的较大下降和第五区段的较大提高。4）小剂量辐照的材料，可溶性蛋白质组分的变化异常，可能与低剂量辐射兴奋效应有关。2.碳离子注入胚乳引起胚的后代发生变异，是典型的重离子诱变产生间接效应的结果。主要表现为：1）M1代的生物学性状、幼苗的抗氧化酶活性、MDA含量、蛋白质含量和染色体畸变率及微核率等发生了较大的变化。2）对其矮杆变异株和紫色茎杆突变体（M2）和RAPD和AFLP分子鉴定等研究，结果显示了矮杆变异体和紫色茎杆突变体的DNA序列发生了改变，而非仅仅产生生理损伤；同一种变异体或突变体，在M2代单株间也存在着明显的差异，表明它们可能还处于分离状态。于是，将M2代以上高代再种，以观察遗传能否稳定。3.重离子辐照普通小麦种子的M1代正常（它们可以完成生长发育的过程）植株同对照材料做正、反交，获得染色体组等来源不同的材料。接着对对照、M2代和正、反交材料DNA进行RAPD和AFLP多态性研究，并将实验结果作统计分析，结果表明：在细胞结构层次上，染色体组作为靶物质，产生的直接效应主要引起随机突变；而非染色体组的细胞组成的间接效应主要引起非随机突变。