芍药栽培品种ISSR反应体系的优化和应用

以栽培芍药为材料,通过对ISSR-PCR反应体系中的Mg~(2 )、dNTP和Tag DNA聚合酶浓度的优化,以改进芍药的ISSR反应体系;从55对引物中筛选出4对有稳定扩增结果的引物;用这4对引物对22个芍药栽培品种的DNA进行扩增,其所得扩增产物与所选芍药性状经过数据处理后,用MATLAB软件分别进行聚类分析,结果表明供试芍药品种皆可分为3个类群,其分子标记与性状的聚类分析间有较强的相关性。     

水稻维生素C合成相关基因的表达分析

根据拟南芥中参和维生素C合成和代谢关键酶的氨基酸序列,经在公共数据库中搜索比对获得了水稻中参与维生素C合成相关基因的信息.在此基础上,利用实时定量PCR技术对这些基因在水稻不同组织器官以及发育种子中的表达进行分析.结果表明:水稻维生素C合成相关基因在叶中的表达量最高,其次是叶鞘和籽粒,茎和根中的表达量最低,而幼嫩叶片中...     

Molecular-Cytological Identification and Chromosome Behavior Analysis of Telotetrasomic in Rice

From the progenies of a telotrisomic of chromosome 9 short arm of an indica rice variety, Zhongxian 3037, a phenotypical variant was selected. The variant plant had rolled leaves, dispersed plant type, as well as a low seed-setting rate. Cytological and molecular cytological investigations revealed two extra chromosomes, which were the shortest in somatic cells of the variant. Fluorescent in situ hybridization (FISH) analysis using a rice centromere specific DNA (RCS2) and a DNA sequence specific for chromosome 9 on premetaphase and pachytene chromosomes showed that these two chromosomes were the short arms of chromosome 9. That is to say, the variant was a telotetrasomic of chromosome 9. Among the 25 pachytene cells, the two telosomic chromosomes paired each other to form a bivalent and didn't pair with other normal chromosome 9 as multivalents in 96% cells. However, the bivalent was easy to disassociate in advance.     

不同转化方法培育无抗性选择标记转基因水稻效率的比较

 为研究不同的转化方法培育无抗性选择标记转基因水稻的效果,以4个籼粳稻品种为受体材料,比较了用农杆菌介导的双菌双载体和双T DNA单载体两种共转化技术的共转化频率及随后获得的无抗性选择标记基因转化子的效率。结果表明,在双菌双载体介导的转化中,4个水稻品种的平均共转化频率为10.1%,其中55.6%的共转化植株自交后代中可筛选出无抗性选择标记的转基因水稻植株。在单载体双 T DNA介导的共转化中,平均共转化频率为45.0%,从其中600%的共转化植株自交后代中可获得无抗性选择标记的转基因植株,即双T DNA单载体法的去选择标记的效率（270%）要较双菌双载体法（5.6%）高。     

Breeding of Selectable Marker-Free Transgenic Rice Lines Containing AP1 Gene with Enhanced Disease Resistance

In order to obtain marker-free transgenic rice with improved disease resistance, the AP1 gene of Capsicum annuum and hygromycin-resistance gene （HPT） were cloned into the two separate T-DNA regions of the binary vector pSB130, respectively, and introduced into the calli derived from the immature seeds of two elite japonica rice varieties, Guangling Xiangjing and Wuxiangjing 9, mediated by Agrobacterium-mediated transformation. Many cotransgenic rice lines containing both the AP1 gene and the marker gene were regenerated and the integration of both transgenes in the transgenic rice plants was confirmed by either PCR or Southern blotting technique. Several selectable marker-free transgenic rice plants were subsequently obtained from the progeny of the cotransformants, and confirmed by both PCR and Southern blotting analysis. These transgenic rice lines were tested in the field and their resistance to disease was carefully investigated, the results showed that after inoculation the resistance to either bacterial blight or sheath blight of the selected transgenic lines was improved when compared with those of wild type.     

抗水稻纹枯病遗传研究的改良方法及其验证

 报道了抗水稻纹枯病遗传研究的一种新方法体系，包括采用新的接种方法、评定病级指标和适宜调查时间，采用永久性遗传分析群体设置有重复的田间试验，田间保持中肥水平，以及浅水灌溉不脱水等内容。用F2无性系群体和重组自交系群体，对新方法进行了4年的验证试验，证明新方法有相当好的重演性。     

Efficiencies of Generating Selectable Marker-Free Transgenic Rice with Different Transformation Methods

To study the efficiency of generating selectable marker-free (SMF) transgenic rice, two transformation methods were employed for four rice varieties (Wuxiangjing 9, Longtefu, Xieqingzao and Zhenshan 97). One method is by using a single twin T-DNA binary vector pYH592 in one Agrobacterium strain, which is composed of two separate T-DNA regions (one carrying an antisense Wx gene and the other carrying a HPT gene). The other one, named as two-strain/ two-vector system, is by using two separate binary vectors in two separate Agrobacterium cultures. The results indicated that the average co-transformation frequencies of the antisense Wx gene and the HPT gene were 10.1% and 45.0%, respectively, for the four rice varieties. And the SMF transgenic plants selected from the offsprings of co-transformants were 55.6% and 60.0% in the two-strain/two-vector and twin T-DNA vector binary systems, respectively.     

水稻异淀粉酶基因ISA1及其启动子的表达特性分析

 利用实时定量RT-PCR方法研究了水稻异淀粉酶基因ISA1在各组织及不同发育阶段籽粒中的表达模式,结果表明该基因只在发育籽粒中表达。同时,克隆了ISA1基因起始密码子上游1.1 kb和2.1 kb两个不同长度的启动子片段,并分别与GUS报告基因融合,经农杆菌介导转入水稻中。对转基因水稻植株中GUS活性的定性与定量结果表明,2.1 kb的ISA1启动子具有很好的胚乳表达特异性,与内源基因定量表达分析的结果一致;而1.1 kb的ISA1启动子则不具备该表达特性,它在胚乳、茎、茎节及谷壳中都有很高的表达。这可能是因为 2.1 kb启动子中含有抑制目的基因在茎等组织或器官中表达的顺式作用元件。     

红莲型不育细胞质在杂交粳稻育种中的应用

为了探讨红莲型不育细胞质应用于杂交粳稻育种的可能性,利用包台(BT)型、红莲(HL)型、茶野(CL)型、野败(WA)型等4种细胞质的同核异质不育系六千辛A与181个粳稻亲本测交,根据测交F1的小穗育性,筛选HL型粳稻恢复系。在此基础上,对筛选出的HL型粳稻恢复系与更多HL型不育系复测,并进行不同细胞质杂交粳稻的比对试验。研究结果表明:1)HL型和CL型不育系的恢、保关系十分一致,测交F1小穗育性间的相关性达极显著水平。2)HL型和CL型粳稻不育系的可恢性虽不如BT型不育系,但明显优于WA型粳稻不育系;以测交F1小穗育性达到85%以上作为选择标准,从BT型恢复系和广亲和恢复系中筛选出25个HL型(CL型)粳稻恢复系。3)不育系的核背景对杂种育性有影响,HL型六千辛A可恢性最好,其次为HL型陵香A,HL型珍5A可恢性最差。4)与BT型不育系配制的杂种相比,HL型和CL型不育系配制的杂种育性稳定性相对较差。5)HL型不育系配制的杂种与BT型、WA型不育系配制的杂种在播种至抽穗历期、株高、产量及品质性状上均无明显差异;HL型不育系配制的杂交粳稻结实率正常,与BT型不育系配制的杂种无明显差异,明显高于WA型不育系配制的杂种。说明HL型不育细胞质应用于杂交粳稻育种是可行的。