中籼杂交水稻亲本多态性的AFLP分析

对15个籼型杂交水稻亲本进行AFLP分析,结果表明：亲本间遗传距离小,在0.0589~0.3305之间,平均为0.2033。15个亲本按类平均法可聚为两类,Ⅰ类为不育系,Ⅱ类为恢复系。其中Ⅱ类又分为两个亚类,Ⅱ-1不含明恢63血缘、Ⅱ-2全部含明恢63血缘。Ⅰ/Ⅱ-1与Ⅰ/Ⅱ-2间的遗传距离无明显差异,揭示恢复系的遗传基础较一致,这可能是当前的品种不能超过汕优63的重要原因之一。要提高水稻的杂种优势,需丰富亲本的遗传基础,扩大其遗传差异。 Abstract:The polymorphisms were analyzed in 15 semilate indica parental materials with AFLPs.The genetic distances between parental materials were small with an average of 0.2033,ranged from 0.0589 to 0.3913.Fifteen parental materials were classified into two groups,cytoplasmic male-sterile lineⅠand restorer line Ⅱ.The latter were classified into two subgroups,Ⅱ-1 and Ⅱ-2.SubgroupⅡ-2 was consanguinity with Minghui63.The genetic distances between male-sterile line and two restorer line subgroups(Ⅱ-1 and Ⅱ-2) did not show significant difference,indicating that the genetic bases of restorer lines were similar,which maybe one of major reasons for the yield still not surpassing Shanyou63 in indica hybrid rice presently.To increase the heterosis of hybrid rice,we must enrich the genetic diversity and expand the genetic differences between parental lines.     

水稻空间技术育种的研究

为探讨空间条件对水稻育种的影响,利用返回式卫星搭载水稻干种子,返地种植,考察分析了12个亲本品种后代的性状表现及遗传与变异情况。结果表明：空间条件诱变的后代性状具有多样性变异和多向性遗传,且能稳定遗传,并有超亲遗传现象,诱变后代具有变异类型多、变异频率高、特殊突变体、育种周期短等特点。为进一步开发利用空间技术育种,在研究实践的基础上,提出了程序化、规范化、可操作性强的水稻空间育种技术规范。 Abstract:In order to inquire into the influences of space conditions on rice breeding,the dry seeds of 12 rice varieties were carried by recoverable satellite.After recovery,these seeds were sowed in the yield,the inheritance and variation of the characters of their progenies were observed and analysed.The results showed that the characters of the progenies mutated under space conditions segregated and varied in many adpects and directions,and the segregated and varied characters were heritable.These progenies posoessed significant transgressive inheritance,a plenty of variation types.and variation.was characterized by short breeding period.The resulfs in this study indicated that space technology breeding could be developed and used as a new method of mutation breeding.     

小麦耐盐突变体的分子生物学鉴定

利用F1花药培养、EMS诱变和耐盐性反复筛选后已稳定9代的小麦耐盐突变体RH8706-49、H8706-34、H8706-44、H8706-48、H8706-57及其亲本濮农3665、百农3039为材料,用生化标记（醇溶蛋白）及分子标记（RAPD）分析了各材料间的差异,发现突变体与亲本相比,不仅发生了蛋白质水平的变异,而且也在DNA水平上证明了突变的发生,从而为耐盐突变体的真实性提供了有力的证据,排除了盐适应的可能性; 经用218 个引物对5个突变体之间的多态性进行RAPD分析,结果表明,它们之间的差异很小,其遗传背景相似,因而它们是一系列耐盐性不同的近似等位基因系。 Abstract:　In this paper, 5 wheat salt tolerant mutants（H8706-34、H8706-44、H8706-48、RH8706-49、H8706-57）derived from anther culture、EMS induction and salt tolerance selection and their parents （Punong3665、Bainong3039）were used as materials, all the mutants have inherited stably for 9 generations. Differences were revealed between the mutants and their parents using chemical marker（gliadin）and molecular marker（RAPD）, the results showed that compared with the parents, the mutants varied not only on the protein level,but also on the DNA level,which supplied hard evidence of the truth of the mutants and ruled out the possibility of salt adaptation. RAPD analysis were conducted among the 5 mutants by 218 primers,which proved they were a series of near isogenic lines of different salt tolerance because of their little difference and similar genetic background.     

一种筛选拟南芥突变体的有效方法

经甲基磺酸乙酯（EMS）诱变处理的拟南芥种子,接种于MS培养基上,垂直放置培养4天后,将幼苗转移至胁迫培养基中,以倒置幼苗180°所形成的弯曲生长根作为指标筛选拟南芥耐营养胁迫突变体。利用这种方法,成功地筛选到一个耐低钾的隐性单基因拟南芥突变体。本方法同样适用于其他类型突变体的筛选。 Abstract：his paper introduces a root-bending assay for isol ation of Arabidopsis mutants tolerant to nutrition stress. Seeds of wild-ty pe Arabidopsis thaliana (ecotype Landersberg erecta) were mutagenized wi th ethyl methyl sulfide (EMS),and M2 populations were screened for mutants. Fo ur-day-old seedlings with 1-to 1.5-cm-long roots were transferred from the vertical agar plates onto to a second agar medium that was supplemented with det erminate stress. The seedlings were arranged in rows, and the plates were orient ed vertically with the roots pointing upward. After another 4 days, the root be nding seedlings were selected for putative mutants and transferred to soil to gr ow to maturity.Seeds from the putative mutants were screened again to determine the true mutants.By using this root-bending assay we have isolated a low-K+-tolerant (lkt1) mutant which is caused by single recessive nuclear mutation. F or lkt1 mutant screening,K+concentration of the medium was 100μmol/L because root growth of wild type seedlings was completely inhibited at or below this con centration.This root-bending assay is also applicable to other type of Arabid opsis mutant isolation.     

空间诱变创造克服灿粳杂种半不育性新种质和新恢源研究

利用高空气球搭载 ,使 2个对胞质雄性不育育性保持的粳稻品种“中作 59”和“海香”干种子处于海拔高度 3 0～ 3 5Km的高宽 8h后回收种植 ,从 SP2 代群体中 ,获得二类突变体 :一类为粳型亲籼并对胞质雄性不育具恢复能力的突变体 ,其突变频率依次为 8× 1 0 -3 和 6× 1 0 -3 。这类突变体的后代稳定株系与典型籼稻品种杂交 F1的结实率在 70 %以上 ,但与典型粳稻品种杂交 F1的结实率只有 3 0 %左右 ,与籼、粳不育系交配 F1的结实率也是如此。另一类为亲籼、粳并对胞质雄性不育具恢复能力的突变体 ,其突变频率依次为 3× 1 0 -3 和2× 1 0 -3 。这类突变体的后代稳定株系与典型籼、粳品种杂交 F1结实率均在 70 %以上 ,与籼、粳不育系测定 F1的结实率也是如此。从这二类突变体后代选出的生育期不同、综合农艺性状和米质均好的粳型亲籼恢复系和亲籼、粳恢复系 ,经 3年 6季与多个籼型不育系测交 F1,不仅表现出明显的籼粳杂种优势 ,而且结实率达 80～ 90 % ,籽粒充实度良好 ,千粒重在 2 3～ 3 2 g之间。本项研究结果表明 ,利用空间诱变创造新恢源和改变粳稻亲和性 ,克服籼粳杂种 F1的不孕性和籽粒充实度差的难题 ,是一条可行的新途径。     

一种紫色水稻的遗传及其在光敏不育系育种中应用的研究

研究表明,本院获得的一种紫色水稻的植株色遗传受控于C、 A、Pl3个独立基因座位上显性基因的互补作用,另有一独立的显性基因对Pl基因的表达起抑制作用。由于该抑制基因在籼稻中的高频率存在,因而,紫稻与一般绿稻品种杂交F1多表现为绿株。紫稻光敏不育(株)系的不育性表达和配合力表现,均可达到与普通光敏不育系相似的水平。本文还讨论了选育籼型紫稻光敏不育系设想的可行性。 Abstract:The inheritance of a purple rice in crosses to green rices was investigated.A group of dominant and interactive genes,C,A and Pl,was found to control the expression of the trait and the other independent inhibitor I-Pl-1 to inhibit the effect of the gene Pl.Because of the wide existence of the gene I-Pl-1 in indica rice,most of F1 plants of the crosses between purple rice and green cultivars were green.The primary study indicated that,for the purple photoperiod-sensitive genic male-sterile lines,the degrees of the sterility and its stability in the sterile stage,and of the fertility in the fertile stage,and the combining ability levels of the purple rice were as high as the degrees and levels for the green rice.A tentative idea on breeding purple photoperiod-sensitive male-sterile lines of indica rice was suggested and its feasibility and advantage was discussed.     

Diversity and stability study on rice mutants induced in space environment

To further study the characteristics of changes on the molecular level of rice mutants induced in space environment, we analyzed proteins in leaves and seeds of four rice mutants （two high-tillering and two low-tillering） in the 8th and 9th generations after a 15-day spaceflight, and compared with their ground controls by two-dimentional polyacrylamide gel electrophoresis and reverse phase liquid chromatography （RPLC）. In addition, the albumin, globulin, prolamine, glutelin, and amylose of the mutant seeds were analyzed by RPLC and ultra-violet spectrometry. The results showed that the low-abundance proteins of leaves in the peak tillering stage are more likely to he induced compared with their corresponding controls. The albumin, globulin, and prolamine of the mutant seeds revealed changes when compared with their controls, and the characteristics of changes in different mu- tants were stably inherited in the 8th and 9th generations, suggesting that they can be used as biomarkers to identity the mutants induced by spaceflight. Moreover, two proteins （SSP9111 and SSP6302） were found to be expressed with high intensity （two-fold change） in different mutants, which were both correlated with photosystem according to mass spectrometry and database searching.     

Protein Interaction Between p53 and △113p53 Is Required for the Anti-Apoptotic Function of △113p53

Zebrafish △113p53, an N-terminal truncated p53 isoform, is a p53-target gene that antagonises p53-mediated apoptotic activity.Interestingly, △113p53 does not act on p53 in a dominant-negative manner, but rather interferes with the p53 function by differentially modulating p53-target gene expression to protect cells from apoptosis. Previous studies showed that over-expressed △113p53 and p53proteins formed a complex. However, it is not known whether endogenous p53 and △113p53 proteins also interact with each other, and if this interaction is required for △113p53 to inhibit the apoptotic activity of full-length p53. In this study, we used two available zebrafish p53 antibodies to address these questions. One, Zfp53-N, only recognises full-length p53, whereas the other, Zfp53-A7C10, detects both full-length p53 and △113p53. Using Zfp53-N for immunoprecipitation and Zfp53-A7C10 for detection, we demonstrated that endogenous △113p53 and full-length p53 induced by a DNA-damaging drug formed a complex in vivo. Furthermore, of the six △113p53 mutants we generated with different point mutations in the oligomerisation domain, two failed to interact with p53 and lost the ability to modulate p53-target gene expression and inhibit p53-induced cell apoptosis. However, those △113p53 mutants that could interact with p53 retained the ability to antagonise the apoptotic activity of p53. Therefore, our data demonstrated that proteineprotein interaction between △113p53and p53 is essential for the anti-apoptotic function of △113p53. In addition, the two △113p53 mutants that failed to interact with p53 are also useful for the study of the mechanisms of other functions of △113p53.     

Protein Interaction Between p53 and △113p53 Is Required for the Anti-Apoptotic Function of △113p53

Zebrafish △113p53, an N-terminal truncated p53 isoform, is a p53-target gene that antagonises p53-mediated apoptotic activity. Interestingly, △113p53 does not act on p53 in a dominant-negative manner, but rather interferes with the p53 function by differentially modulating p53-target gene expression to protect cells from apoptosis. Previous studies showed that over-expressed △113p53 and p53 proteins formed a complex. However, it is not known whether endogenous p53 and △113p53 proteins also interact with each other, and if this interaction is required for △113p53 to inhibit the apoptotic activity of full-length p53. In this study, we used two available zebrafish p53 antibodies to address these questions. One, Zfp53-N, only recognises full-length p53, whereas the other, Zfp53-A7C10, detects both full-length p53 and △113p53. Using Zfp53-N for immunoprecipitation and Zfp53-A7C 10 for detection, we demonstrated that endogenous △113p53 and full-length p53 induced by a DNA-damaging drug formed a complex in vivo. Furthermore, of the six △113p53 mutants we generated with different point mutations in the oligomerisation domain, two failed to interact with p53 and lost the ability to modulate p53-target gene expression and inhibit p53-induced cell apoptosis. However, those △113p53 mutants that could interact with p53 retained the ability to antagonise the apoptotic activity of p53. Therefore, our data demonstrated that protein--protein interaction between △113p53 and p53 is essential for the anti-apoptotic function of △113p53. In addition, the two △113p53 mutants that failed to interact with p53 are also useful for the study of the mechanisms of other functions of △113p53.     

杂交水稻产量性状配合力研究

利用 9个不育系和 7个恢复系配制 40个杂交组合 ,对 6个产量性状的亲本一般配合力和组合特殊配合力效应进行分析。结果表明 ,杂种一代 (F1)各性状的形成同时受亲本一般配合力和组合特殊配合力的影响。单株穗数、每穗总粒数、千粒重三性状主要受一般配合力作用 ;每穗实粒数、结实率、单株粒重三性状则一般配合力和组合特殊配合力的作用同样明显 ,或特殊配合力作用更明显些。不育系珍汕 97A、龙特浦A、K1 8A和恢复系明恢 63、直龙、1 0 2 5的一般配合力高 ,利用它们容易配制出高产组合。两年试验结果千粒重、单株粒重二性状的形成均是不育系的作用大于恢复系 ,近期育种工作重点应放在不育系的选育上。     

一个CMS水稻育性回复突变体的RAPD分析

以籼稻细胞质雄性不育系Ⅱ－３２Ａ、保持系Ⅱ－３２Ｂ、恢复力由单基因控制的育性回复突变体（Ｇ３和Ｅ１５）,以及两个恢复系（明恢６３和ＩＲ２４）的基因组ＤＮＡ为材料,用ＲＡＰＤ方法比较分析了各材料间的差别和亲缘关系。发现可育回复突变体和不育系间有一定的差异,但远远小于两个恢复系和不育系间的差异,甚至小于保持系和不育系间的差异,从而在ＤＮＡ水平上为育性回复突变体的真实性提供了有力的证据,排除了机械混杂或惭复系串粉的可能性。实验也说明辐射可引起基因组内广泛的变异,但大部分遗传的变异并不影响生物的性状和生活力。另外发现突变体中出现的多态片段有二部分也存在于其他品种中,说明辐射引起的突变在整个水稻种群中并不一定是全新的,有的可能属于回复突变     

Features of the expressed sequences revealed by a large-scale analysis of ESTs from a normalized cDNA library of the elite indica rice cultivar Minghui 63

The indica subspecies of cultivated rice occupies the largest area of rice production in the world. However, a systematic analysis of cDNA sequences from the indica subspecies has not been performed. The aim of the present study was to collect and analyze the expressed sequence tags (ESTs) of indica rice on a large scale. A total of 39 208 raw sequences were generated from a normalized cDNA library prepared by use of 15 different tissues of the indica cultivar Minghui 63. After trimming, processing and analysis, 17 835 unique sequences were obtained, each of which presumably represents a unique gene. Of these sequences, 2663 were novel, and at least 70 were indica specific. Comparison of the Minghui 63 sequences with the ESTs/full-length cDNAs in GenBank revealed a large number of deletion/insertion/substitution (DIS) at both the inter- and intra-subspecific levels. The overall number of polymorphisms in the expressed sequences was higher in the inter-subspecific comparisons than in the intra-subspecific comparisons. However, the extent of DIS-based polymorphism was highly variable among different rice varieties. In total, 15 726 unique sequences, including 697 novel sequences, were assigned to regions where large numbers of quantitative trait loci (QTLs) for agronomic traits had been detected previously. These results may be useful for developing new molecular markers for genetic mapping, detecting allelic polymorphisms associated with phenotypic variations between rice varieties, and facilitating QTL cloning by providing the starting points for candidate-gene identification.     

水稻转基因系"明恢63-Xa21"的基因组分析

植物基因工程研究已经建立了多种转基因的方法 ,这些方法包括农杆菌侵染[1 ] 、粒子轰击[2 ] 、电激转化和原生质体培养[3 ] 等。研究人员希望通过这些方法 ,将功能外源基因整合到受体基因组 ,而同时不引起其它性状的变化。已有的研究表明 ,各种转基因系统均能成功地将外源基因整合到受体基因组并能稳定地遗传到后代[1～ 3 ] 。然而通常情况下人们主要关注目标性状的变化 ,而对受体基因组的其它变化研究较少。事实上许多转基因植物发生了不希望出现的变异[4,5] 。已建立的各种分子标记如ＳＳＲＰ(简单序列重复多态性 ) [6] 、ＲＡＰＤ(随机…     

Expression profile analysis of the polygalacturonase-inhibiting protein genes in rice and their responses to phytohormones and fungal infection

Polygalacturonase-inhibiting proteins (PGIPs) are typically leucine-rich repeat (LRR) proteins that can inhibit the activity of fungal polygalacturonases (PGs). In this study, two new Ospgip genes, named Ospgip6 and Ospgip7 with consensus sequence of ten imperfect LRR motif located on rice chromosomes 8 and 9, were identified using BLAST analysis. Both of them appear to be extracellular glycoproteins. To have a global view of the dynamic gene expression pattern, seven Ospgip genes were first analyzed using the Affymetrix rice genome array data from online resource. All of these seven Ospgip genes showed variable expression patterns among tissues/organs. In order to further investigate the potential function of these Ospgip genes, the responses of Ospgip genes to the treatment of various phytohormones (abscisic acid, brassinosteroid, gibberellic acid, 3-indole acetic acid, jasmonic acid, kinetin, naphthalene acetic acid and salicylic acid) as well as fungal infection were analyzed by real-time PCR using time course array. Generally, all the Ospgip genes were slightly up-regulated in the indica rice cultivar Minghui 63 under GA(3), KT and NAA treatments (except Ospgip2, which was down-regulated under KT treatment). In the japonica rice cultivar Zhonghua 11, Ospgip genes were regulated by most treatments with the response time variability. We also analyzed putative cis-elements in the promoter regions of Ospgip genes. This dataset provided a versatile resource to understand the regulatory network of Ospgip genes during the process of phytohormones treatment and fungal infection in the model monocotyledonous plant, rice, and could aid in the transgenic breeding against rice fungal diseases. KEY MESSAGE: All the seven Ospgip genes showed variable expression patterns in Minghui 63 and their expressions were regulated by different phytohormone treatments or fungal infection in Minghui 63 and Zhonghua 11.     

通过转PSAG12—IPT基因培育延缓叶片衰老水稻

利用农杆菌介导的遗传转化方法将PSAG12 _IPT导入籼稻品种明恢 6 3。在获得的 6 1个独立的转基因植株中 ,有一些表现出叶片衰老显著延缓 ,对选择的两个纯合转基因株系的小区试验的结果显示 :(1)转基因植株倒三叶的持绿性极显著延长 ;(2 )两个纯合转基因株系比原品种的结实率极显著提高 ,株高极显著降低 ;(3)两个纯合转基因株系的有效穗数比原品种极显著或显著提高。     

Field performance of transgenic elite commercial hybrid rice expressing bacillus thuringiensis delta-endotoxin

Here we describe development of transgenic elite rice lines expressing a Bt fusion gene derived from cryIA(b) and cryIA(c) under the control of rice actinI promoter. The lines used in the study were indica CMS restorer line of Minghui 63 and its derived hybrid rice Shanyou 63. The level of Bt fusion protein CryIA(b)/CryIA(c) detected in Minghui 63 (T51-1) plants was 20 ng/mg soluble protein. The Bt Shanyou 63 was field-tested in natural and repeated heavy manual infestation of two lepidopteran insects, leaffolder and yellow stem borer. The transgenic hybrid plants showed high protection against both insect pests without reduced yield.     

Comparison of quantitative trait loci controlling seedling characteristics at two seedling stages using rice recombinant inbred lines

A better understanding of the genetics of seedling characteristics in rice could be helpful in improving rice varieties. Zhenshan 97 and Minghui 63, the parents of Shanyou 63, an elite hybrid developed during the last decade in China, vary greatly with respect to their physiological and morphological traits at the seedling growth stage. In this study, we used a population of 240 recombinant inbred lines derived from a cross between Zhenshan 97 and Minghui 63 to identify quantitative trait loci (QTL) for seedling characteristics. All plant material was grown in hydroponic culture. Data for the following characters were collected at 30 days and 40 days post-sowing: plant height, shoot dry matter weight (SDW), maximum root length, root dry weight (RDW), total dry weight , and root-shoot ratio (the ratio of SDW to RDW). Analysis using composite interval mapping detected 16 QTL for the six traits in 30-day-old seedlings. Of these 16 QTL, Minghui 63 alleles increased trait values at only two of them. The QTL in the vicinity of R3166 on chromosome 5 simultaneously influenced PH, SDW, MRL, RDW, and TDW in the same direction. Twenty QTL were detected for the same traits in the 40-day-old seedlings. However, at this stage Minghui 63 alleles increased trait values at eight QTL. The QTL linked to R3166 also affected PH, SDW, MRL, RDW, and TDW. Only four QTL were common to the two stages. These results clearly indicate that different genes (QTL) control the same traits during different time intervals. Zhenshan 97 alleles had positive effects during the first 30 days of seedling growth, but thereafter the positive effects of Minghui 63 alleles on seedling growth gradually became more pronounced.     

利用SSR标记定位明恢63的2对恢复基因

选取珍汕97A和明恢63杂交组合的F2高可育和高不育单株构建基因池,利用302对SSR引物对其进行了多态性分析。结果表明,位于第1染色体上的RM1和位于第10染色体上的RM258,RM304在亲本,基因池间表现多态性,用F2单株验证证明它们与野败型恢复基因连锁,完全不育株分析表明,与恢复基因间的遗传距离分别为1.9,2.9和0.0cM,野败型,红莲型,BT型3种不育胞质恢复基因在第10染色体上可能为同一基因或家族成员。     

不同水稻品种上白背飞虱取食行为的EPG分析

【目的】为探明不同水稻品种（丰源优272、R9810-T、华恢1号、明恢63、麻糯谷和Rathu Heenati）对白背飞虱Sogatella furcifera的抗性差异机理,利用刺吸电位技术(electronic penetration graph, EPG)记录了该虫在这6个品种三叶期稻苗上的取食行为。【方法】结合特定的EPG波形,考察了9个非韧皮部指标和22个韧皮部指标。【结果】在总记录时间8 h内,白背飞虱在品种Rathu Heenati上非刺探波np的总时间最长,其次是麻糯谷上非刺探波np的总时间,这两者间差异显著且都显著地长于其他4个品种的np总时间(P<0.05);该虫在Rathu Heenati上路径波Nc的总时间为8 523.41 s,是感虫品种明恢63 Nc总时间的2.24倍;该虫在Rathu Heenati上吸食韧皮部汁液N4-b的总时间显著地比其他品种短(P<0.05)。华恢1号和R9810-T上所有的EPG指标和感虫品种明恢63的EPG指标都没有显著的差异(P≥0.05)。白背飞虱在丰源优272上单次分泌水溶性唾液N4-a的平均时间更长,并伴随长时间的韧皮部取食。【结论】由此推测,品种Rathu Heenati可能存在不利于白背飞虱取食的忌避成分,并且在韧皮部外组织和韧皮部组织都存在抗性因子;在麻糯谷上仅存在忌避成分;然而,品种华恢1号和R9810-T可能对白背飞虱不具有明显的抗性;丰源优272可能是比明恢63更为感虫的品种。结合介体昆虫取食行为与传播持久性病毒的关系,这些结果也为利用抗白背飞虱品种控制南方水稻黑条矮缩病提供了参考。