DNA分子标记在果树种质资源遗传多样性研究中的应用

简要介绍了DNA分子标记的主要类型、原理及特点,重点综述了RFLP、RAPD、AFLP、SSR、EST-SSR和SNP标记技术在果树种质资源遗传多样性研究中的应用现状。     

Identification of molecular markers associated with sugar-related traits in a <Emphasis Type="Italic">Saccharum</Emphasis> interspecific cross

The cultivated sugarcane (Saccharum spp. hybrids, 2n = 100–130) is one crop for which interspecific hybridization involving wild germplasm has provided a major breakthrough in its improvement. Few clones were used in the initial hybridization event leading to a narrow genetic base for continued cultivar development. Molecular breeding would facilitate the identification and introgression of novel alleles/genes from the wild germplasm into cultivated sugarcane. We report the identification of molecular markers associated with sugar-related traits using an F₁ population derived from a cross between S. officinarum ‘Louisiana Striped’ × S. spontaneum ‘SES 147B’, the two major progenitor species of cultivated sugarcane. Genetic linkage maps of the S. officinarum and S. spontaneum parents were produced using the AFLP, SRAP and TRAP molecular marker techniques. The mapping population was evaluated for sugar-related traits namely, Brix (B) and pol (P) at the early (E) and late (L) plant growing season in the plant cane (04) and first ratoon (05) crops (04EB, 04LB, 04LP, 05EB and 05EP). For S. officinarum, combined across all the traits, a total of 30 putative QTLs was observed with LOD scores ranging from 2.51 to 7.48. The phenotypic variation (adj. R²) explained by all QTLs per trait ranged from 22.1% (04LP) to 48.4% (04EB). For S. spontaneum, a total of 11 putative QTLs was observed with LOD scores ranging from 2.62 to 4.70 and adj. R² ranging from 9.3% (04LP) to 43.0% (04LB). Nine digenic interactions (iQTL) were observed in S. officinarum whereas only three were observed in S. spontaneum. About half of the QTLs contributed by both progenitor species were associated with effects on the trait that was contrary to expectations based on the phenotype of the parent contributing the allele. Quantitative trait loci and their associated effects were consistent across crop-years and growing seasons with very few QTLs being unique to the early season. When the data were reanalyzed using the non-parametric discriminant analysis (DA) approach, significant marker-trait associations were detected for markers that were either identical to or in the vicinity of markers previously identified using the traditional QTL approach. Discriminant analysis also pointed to previously unidentified markers some of which remained unlinked on the map. These preliminary results suggest that DA could be used as a complementary approach to traditional QTL analysis in a crop like sugarcane for which saturated linkage maps are unavailable or difficult to obtain.     

Cultivar identification and genetic map of mango (Mangifera indica)

Amplified Fragment Length Polymorphism (AFLP) information was used for identification of mango (Mangifera indica L.) cultivars, for studying the genetic relationship among 16 mango cultivars and seven mango rootstocks and for the construction of a genetic linkage map. Six AFLP primer combinations produced 204 clear bands and on the average 34 bands for each combination. The average Band-Sharing between cultivars and rootstocks was 83% and 80%, respectively. The average Band-Sharing for mango is 81%. The probability of obtaining a similar pattern for two different mango cultivars and rootstocks is 6 × 10⁻³and 2 × 10⁻³, respectively. A preliminary genetic linkage map of the mango genome was constructed, based on the progeny of a cross between ‘Keitt’ and ‘Tommy-Atkins’. This linkage map consists of 13 linkage groups and covers 161.5 cm defined by 34 AFLP markers. This revised version was published online in August 2006 with corrections to the Cover Date.     

亚洲棉种质资源的SSR遗传多样性分析

 对我国棉花中期库保存的200份不同地理来源的亚洲棉代表性样本进行了SSR遗传多样性分析,结果表明：亚洲棉分子水平的遗传多样性较高。83个多态性位点共检测到368个等位基因变异,其中多态性的等位基因数为329个,平均每个SSR位点3.964个。位点多态性信息量（PIC）变幅为0.010～0.882,平均0.578,PIC值大于0.7的标记有33个（占39.8%）。基因多样性（H′）变幅为0.031～2.163,有效等位基因数（Ne）变幅为1.010～8.496。华南棉区基因遗传多样性最高,其次为长江流域棉区、黄河流域棉区,从理论上支持被广泛接受的亚洲棉在我国的传播路线是由南到北,华南棉区是中棉种系的遗传多样性富集中心。利用软件NYSTS-pc2.20,采用类平均法(UPGMA)进行聚类分析,种质间SSR相似系数变幅为0.58～0.997,平均0.745,在阈值0.73处200份亚洲棉聚为8个类群,贵池小子棉白子单独聚为一群,与其他种质遗传距离较远。遗传距离和地理距离没有必然联系,但种质间亲缘关系处于极端远或极端近时,则地理距离一般也趋于较远或较近。     

功能标记及在品种鉴定和辅助育种中的应用前景

功能标记是根据功能基因内部引起表型性状变异的多态性基序开发出来的一种新型分子标记.由于来自基因内的功能性基序,此类标记不需要进一步验证就可以在不同的遗传背景下确定目标等位基因的有无.本文详细描述了功能标记的概念及与其它几种类型遗传标记相比存在的优势:提出了功能标记开发的基本条件和发展策略.利用关联分析方法和近等基因系途径可以获得直接或间接的功能标记.结合本单位的研究方向,探讨了功能标记在分子标记辅助育种和品种鉴定中的应用潜力和开展相关工作的一些设想.     

Molecular tagging of Asiatic soybean rust resistance in exotic genotype EC 241780 reveals complementation of two genes

Asian soybean rust (ASR) caused by Phakopsora pachyrhizi severely reduces seed yield in soybean. Molecular tagging of ASR resistance can help in the process of resistance breeding. In this study, an F₂ population of cross (susceptible cultivar ‘NRC 7’ × resistant exotic genotype EC 241780) was used for bulked segregant analysis (BSA) with 25 SSR (simple sequence repeat) primers linked with six Rpp genes. Among them, five polymorphic SSR markers, viz., Sct 187, SSR 1859, Satt 191 (Rpp1b like loci) and Satt 215, Sat_361 (Rpp2 loci) distinguished the ASR resistant and susceptible bulks and individuals. In combined marker analysis, the markers Satt 191 (Rpp1b like loci) and Satt 215 (Rpp2 loci) were linked with ASR severity score and were also confirmed in individual 110 F₂ segregants. Hence, these markers could be utilized in the marker assisted rust resistance breeding of Rpp1b like and Rpp2 genes. In silico candidate gene analysis for hypersensitive response revealed that Satt 191 linked region was rich in genes encoding apoptotic ATPase having leucine‐rich repeat (LRR) domain.     

Analysis of genetic relationships among 22 European barley varieties based on two PCR markers

Two long primers of 19 (F17) and 20 (F13) nucleotides, respectively,were used in polymerase chain reactions to amplify DNA from differentcultivated barley accessions. These primers can distinguish closely relatedvarieties and, with a unique primer, all the barley accessions analysedshowed a characteristic fingerprint. Sixty per cent and 76% of thefragments generated using F13 and F17, respectively, were polymorphic.The genetic similarity values between accessions were estimated from F13and F17 data. The dendrogram and principal coordinate analysis performedwith F13 data revealed a clear separation of these varieties in accord withtheir pedigree relationships.     

Genetic analysis of a CSR10 (indica) × Taraori Basmati F3 population segregating for salt tolerance using ISSR markers

Segregation for salinity tolerance and ISSR markers based molecular polymorphism were investigated in a F₃ plant population raised via single-seed descent method from a cross between salt-tolerant indica rice variety CSR10 and salt-susceptible premium traditional Basmati rice variety Taraori Basmati HBC19. A total of 130 F₃plants were evaluated individually for salinity tolerance on 1–9 scale on the basis of seedling growth parameters; the average score ranged between 1.7 to 8.3. Frequency distribution curve obtained using the salinity tolerance data of F₃ population and a chi-square analysis, showed a good fit to a normal distribution. Eleven plants each in the category of salt-tolerant and salt-susceptible were selected from the segregating F₃ population for ISSR marker analysis. A total of 149 bands (4–11 bands per primer) ranging from 200 to 3530 bp were scored for the two rice varieties and the selected CSR10 × HBC19 segregating F₃ plants using 26 ISSR primers. Of these, 89 were monomorphic and 60 were polymorphic. Of the 60 polymorphic bands,36 and 20 bands were specific to CSR10 andHBC19 respectively. The remaining four bands were amplified using UBC primers 810,848, 853 and 886 and present in only some of the CSR10 × HBC19 F₃ plants. Notably, ISSR primers with dinucleotide repeat motif and 5'-anchored end amplified more number of bands (7.0 bands/primer) compared to3'-anchored dinucleotide primers (5.4bands/primer), but 3'-anchored dinucleotide primers revealed higher level of polymorphism (2.6 polymorphic bands/primer) compared to 5'-anchoreddinucleotide primers (1.43 polymorphic bands/ primer). While distribution of majority of the polymorphic bands were more or less in the expected ratios in salt-tolerant and/or salt-sensitive F₃segregating plants, but some of the bands amplified using UBC ISSR primers 823, 825,826, 849, 853, 864, 866 and 884 showed highly skewed distribution. Such polymorphic bands stand greater chances of having a linkage with the genes/ QTLs for salinity tolerance and shall be the target for further studies. This revised version was published online in July 2006 with corrections to the Cover Date.     

用RAPD标记研究中国木本植物胶孢炭疽菌的群体分化

收集并鉴定了43个胶孢炭疽菌菌株，用随机扩增多态性DNA(RAPD)研究了胶胞炭疽菌种内的遗传分化。结果表明供试的43个菌株间遗传相似度变化较大。RAPD分子标记将43个胶胞炭疽菌菌株分为2个类群，其种内遗传分化与寄主没有相关性。因此认为根据寄主种类划分林木胶孢炭疽菌种内专化型不合适，用分子生物学方法更准确。     

生物技术在橡胶树育种中的应用

综述了生物技术在橡胶树育种中的应用，主要包括组织培养技术、染色体工程技术、分子标记技术、基因工程技术等方面的内容；并展望了其在橡胶树育种中的前景。