来源于中国春×洛夫林10的DH群体的磷利用效率的鉴定

黑麦(Secale cereale L.)1R染色体上带有磷高效基因,含有1RS/1BL易位的洛夫林10号是-个磷高效的品种.证明P效率的基因是否位于1RS上和洛夫林10号的P高效基因是否来源于1RS具有重要意义.为了验证该假设正确与否,调查了一个DH群体的61个系的P利用效率,该群体来自于高效吸收磷的品种洛夫林10号和低效吸收磷的中国春的F1代花药培养,是否带有1RS/1BL易位和不含有1RS/1BL易位的DH系间的磷利用效率存在差异?应用酸性聚丙烯酰胺凝胶电泳(A-PAGE)和基因组原位杂交(GISH)鉴定DH群体中的1RS/1BL易位系.A-PAGE分析表明61个DH系中有34个含有1RS/1BL易位染色体,因为它们含有1RS特有的Sec-1醇溶蛋白带纹.进一步用GISH证明了34个系中33个含有一对1RS/1BL纯合易位,只有一个是1RS/1BL单体.田间试验在缺磷土壤中进行,调查各DH系以及它们的亲本在-p(不施P肥)和 P(60kg P/hm-2)条件下的籽粒产量、生物量、每株穗数、磷吸收效率和磷利用效率.结果表明,土壤缺磷降低洛夫林10号的前四个性状的值,但中国春的降低更加严重.洛夫林10号无论在 P和-p条件下所有测试性状的值都高于中国春,但磷的利用效率二者相似.在-P和 P条件下,所测定的五个性状存在分离,且在DH系之间有显著差异;虽然DH系间的变异超出双亲,但是所有DH系的平均值介于双亲之间.前述五个性状的平均值和耐低磷值(-P/ P的相对籽粒产量)在含有1RS/1BL易位和无1RS/1BL易位的DH系之间没有差异.这表明在洛夫林10号中的1RS与磷的利用效率和耐低磷能力没有关系,1RS上可能没有磷高效基因.因此,有可能从洛夫林10号的后代中选出高品质、高效利用磷和耐低磷的优良品种.     

119份小麦种质资源醇溶蛋白遗传多样性分析

利用A-PAGE对119份小麦种质资源进行了醇溶蛋白遗传多样性分析。结果显示,共分离出蛋白带1 938条,迁移率不同的谱带类型116种,其中编号为10和14的2种谱带出现频率最高,分别为54.62%和96.44%,其余114种谱带类型具有较强的多态性;每个种质材料的醇溶蛋白谱带数为10~24条,大部分为13~19条;供试种质间遗传距离在0.24~0.83,平均为0.54;聚类分析将所测材料分为6大类,与种质资源所反映的系统关系类似,表明醇蛋白在一定程度上能反映种质间的亲缘关系。     

Identification and Characterization of Phosphorus Use Efficiency in a Doubled Haploid Population of Chinese Spring×Lovrin No.10

It is reported that chromosome 1R of rye (Secale cereale L.) convey phosphorus use efficient gene (s), and 1RS/1BL translocation genotype Lovrin No.10 is P use efficient. So we hypothesized whether P efficient gene(s) locate on 1RS, and the high P efficiency of Lovrin No.10 is from 1RS? To test this hypothesis, we investigated the P use efficiency (PUE) of a doubled haploid (DH) population with 61 lines derived from anther culture of F₁ hybrid between Lovrin No.10 and phosphorus uptake inefficient genotype Chinese Spring to see whether PUE differs between DH line with and without 1RS/1BL translocation. Acidic polyacrylamide-gel electrophoresis (A-PAGE) of gliadin and genomic DNA in situ hybridization (GISH) were employed to discriminate 1RS/1BL translocation DH lines from the normal 1B DH lines. Among the 61 DH lines investigated, A-PAGE analysis showed that 34 lines contained the 1RS/1BL translocation chromosome, which was characterized by the presence of a 1RS-specific Sec-1 marker bands. Further verification with GISH proved that 33 in the 34 lines contained a pair of homozygous 1RS/1BL translocation chromosomes, only one line was a 1RS/1BL monosomic line. A field experiment was carried out on P deficient soil to investigate grain yield, biomass, numbers of spikes per plant (SPP), P uptake efficiency (PUpE), and P utilization efficiency (PUtE) of the DH lines and their parents under -P (nil P applied) and +P (60 kg P/hm² applied) at maturity. Results showed soil P deficiency decreased the values of the first four traits in Lovrin No.10, but were more severe for Chinese Spring. Lovrin No.10 had higher values of all the above tested traits at both -P and +P than Chinese Spring did, but had similar PUtE with Chinese Spring. These five traits segregated, and differed greatly among DH lines under both -P and +P conditions. Although the variations among DH lines exceeded the difference between the two parents, the average values of the DH lines were between the two parents. The average of the above five traits, and P deficiency tolerance (PDT) (measured by relative grain yield of -P/+P) were not different between the DH lines with and without 1RS/1BL translocation. This indicated that there was no association between 1RS and PUE and PDT in Lovrin No.10, and 1RS may not have P efficient gene(s). Therefore, in the offspring of Lovrin No.10, it is possible to combine high PUE and PDT with good quality without the negative effect of 1RS on flour quality.     

冰草属(Agropyron Gaertn.)植物遗传多样性取样策略基于醇溶蛋白的研究

对冰草属(Agropyron Gaertn.)植物的5个种22个居群分8个取样梯度,分别随机取3、6、9、12、15、18、21、24粒种子的混合样进行醇溶蛋白的A-PAGE分析,结果发现:平均Simpson指数依取样量的增加表现增大,以3粒种子0.922表现最低,以21粒0.933表现最高,变异系数(CV%)为0.41.醇溶蛋白谱带数依所提取的混合样的籽粒数目发生显著变化,冰草属内5个种表现趋势相同,22个居群不同取样整体平均以3粒种子提取的醇溶蛋白谱带数18.38为最少,以18粒种子提取的醇溶蛋白谱带28.24为最多,CV%为13.21.取样量达到12粒以上醇溶蛋白谱带差异不明显(P=0.01),图谱表现基本一致.建议在利用生化指纹进行冰草属居群间及种间的遗传多样性研究中,混合取样量最低应保持在12个个体及以上方能代表居群整体,反映居群的整体遗传特性.     

Encoding genes for endosperm proteins in Hordeum chilense

Summary The endosperm proteins encoded by the genome Hch in Hordeum chilense, Tritordeum (amphiploid Hordeum chilense x Triticum turgidum), common wheat-H. chilense addition lines, and the segregating plants resulting from the cross Tritordeum x T. turgidum, were fractionated by three electrophoretical techniques: SDS-PAGE, A-PAGE, and bidimensional PAGE. Prolamin subunits with a high molecular weight (HMW) were well visualized by SDS-PAGE, the A-PAGE technique permitted good resolution for many hordeins and gliadins, and two-dimensional electrophoresis allowed new sets of bands coded by gene complexes from H. chilense chromosomes to be distinguished. The loci Hor-Hch1 (up to 11 subunits belonging to the -, — and -hordeins), Glu-Hch1 (one HMW prolamin subunit), Hor-Hch2 (one -hordein), and Hor-Hch3 (up to four -hordeins) were located on the H. chilense chromosomes 1Hch, 5Hch, and 7Hch.     

K、V、T型小麦细胞质雄性不育系籽粒的醇溶蛋白分析

目的:通过对K、V、T型细胞质雄性不育系,保持系及其杂交种籽粒的醇溶蛋白进行A-PAGE分析,以寻找与细胞质雄性不育基因表达有关的特异蛋白。方法:A-PAGE电泳分析,凝胶浓度6.72%。结论:太911289不育系的电泳图谱与保持系相比在β区缺失一条谱带,但是在冀5418不育系的图谱中并未发现与保持系区别的谱带,这与小麦醇溶蛋白基因表达的品种特异性有关;通过对杂交种图谱的分析发现,杂交种图谱与其父本差异很大,具有很强的偏母性,同时各不育类型的杂交种间也存在一定差异。     

来源于中国春×洛夫林10的DH群体的磷利用效率的鉴定(英文)

黑麦(SecalecerealeL.)1R染色体上带有磷高效基因,含有1RS/1BL易位的洛夫林10号是一个磷高效的品种。证明P效率的基因是否位于1RS上和洛夫林10号的P高效基因是否来源于1RS具有重要意义。为了验证该假设正确与否,调查了一个DH群体的61个系的P利用效率,该群体来自于高效吸收磷的品种洛夫林10号和低效吸收磷的中国春的F1代花药培养,是否带有1RS/1BL易位和不含有1RS/1BL易位的DH系间的磷利用效率存在差异?应用酸性聚丙烯酰胺凝胶电泳(A-PAGE)和基因组原位杂交(GISH)鉴定DH群体中的1RS/1BL易位系。A-PAGE分析表明61个DH系中有34个含有1RS/1BL易位染色体,因为它们含有1RS特有的Sec-1醇溶蛋白带纹。进一步用GISH证明了34个系中33个含有一对1RS/1BL纯合易位,只有一个是1RS/1BL单体。田间试验在缺磷土壤中进行,调查各DH系以及它们的亲本在-P(不施P肥)和 P(60kgP/hm-2)条件下的籽粒产量、生物量、每株穗数、磷吸收效率和磷利用效率。结果表明,土壤缺磷降低洛夫林10号的前四个性状的值,但中国春的降低更加严重。洛夫林10号无论在 P和-P条件下所有测试性状的值都高于中国春,但磷的利用效率二者相似。在-P和 P条件下,所测定的五个性状存在分离,且在DH系之间有显著差异;虽然DH系间的变异超出双亲,但是所有DH系的平均值     

高粱属A-PAGE 图谱的建立及在种子鉴定中的应用

以高粱、苏丹草等5种高粱属植物的种子为材料,利用酸性聚丙烯酰胺凝胶电泳(A-PAGE)检测其醇溶蛋白.同时研究了丙酮处理、样品上样量和凝胶浓度对电泳的影响.结果表明:样品经丙酮处理后,在上样量为32μg、凝胶质量分数为15%时电泳可以得到清晰的A-PAGE图谱,据此可以鉴定5种高粱属植物.     

中药桑寄生凝集素的分离及体外抗肿瘤活性的研究

采用硫酸铵分级沉淀和酸化的Sepharose-4B亲和层析分离得到桑寄生凝集素;通过A-PAGE和SDS-PAGE电泳分析,确定所得凝集素组分和亚基情况;通过MTT法对所得到的凝集素进行体外抗肿瘤活性研究。研究结果表明,桑寄生凝集素包含两种组分,共含有四个亚基,亚基分子量分别为36.8 kDa、33.4 kDa,31.3 kDa和29.3 kDa;凝集素具有抗肿瘤效果,对肝癌BEL-7402细胞和胃癌MGC-823细胞的IC50分别为24.2μg/mL和20.9μg/mL。     

应用A-PAGE技术研究玉米自交系资源种子蛋白的遗传多样性

采用酸性聚丙烯酰胺凝胶电泳(A-PAGE)技术对260份玉米自交系种质资源的种子储藏蛋白进行检测,分析了这组种质在蛋白水平上的遗传多样性。结果表明,在供试自交系中,种子盐溶和酸溶蛋白电泳谱带均显示出较高的多态性,并存在一致性。其中盐溶蛋白电泳谱带数目多于酸溶蛋白,且95%为多态性谱带,表明盐溶蛋白在供试自交系资源中具有丰富的多样性。利用盐溶蛋白多态性完成的聚类分析结果和自交系系谱及杂种优势群之间没有明显的对应关系,表明储藏蛋白A-PAGE方法对于大规模玉米种质资源的遗传多样性分析和类群划分可能存在局限性。