棉花黄萎病抗性的分子研究进展

本文综述了棉花黄萎病菌的分子鉴定，棉花黄萎病抗性的遗传基础，分子标记在棉花抗黄萎病方面的应用，棉花黄萎病抗性基因的克隆及转化等方面的研究进展。     

大豆基因组SSR和EST-SSR在黄芪中的通用性分析

分子标记的种间通用性可降低其开发成本,提高利用效率,促进遗传研究较薄弱物种的分子遗传学研究。为开发黄芪SSR分子标记,本研究利用大豆的66对基因组SSR(G-SSR)和43对EST-SSR引物在黄芪基因组中进行通用性分析,并选出其中条带清晰易辨的23对引物对6种不同来源的黄芪进行遗传分析。结果表明:大豆G-SSR、EST-SSR引物在黄芪中的通用性比例分别为31.82%、76.74%,多态性引物分别占18.18%、53.49%,大豆EST-SSR在黄芪的通用性高于基因组SSR,且通用的大豆SSR标记可以用于不同来源黄芪的遗传多样性分析。本研究发掘的多态性SSR引物可以有效用于黄芪的分子遗传多样性研究。     

大白菜抗根肿病基因位点CRa和CRb的分子标记鉴定

利用大白菜抗根肿病基因CRa和CRb分子标记(SC2930和KBr H129J18R)引物组,对78份大白菜材料进行抗根肿病分子标记鉴定。结果表明,在这78份材料中,有34份材料含有SC2930-T(CRa抗病标记)标记,其中杂合抗病位点材料17份,纯合抗病位点材料17份。有37份材料含有KBr H129J18R抗病标记,其中纯合抗病位点材料15份,杂合抗病位点材料22份。有20份材料不含有CRa和CRb所对应的抗病标记,23份材料含有2个抗病标记。该研究初步明确了78份参试大白菜材料所含抗根肿病基因CRa和CRb类型,为大白菜抗根肿病育种提供材料选择依据。     

Fusarium wilt of chickpea: physiological specialization, genetics of resistance and resistance gene tagging

Chickpea wilt caused by Fusarium oxysporum f. sp. ciceris is one of the major yield limiting factors in chickpea. The disease causes 10–90% yield losses annually in chickpea. Eight physiological races of the pathogen (0, 1A, 1B/C, 2, 3, 4, 5 and 6) are reported so far whereas additional races are suspected from India. The distribution pattern of these races in different parts of the world indicates regional specificity for their occurrence leading to the perception that F. oxysporum f. sp. ciceris evolved independently in different regions. Pathogen isolates also exhibit differences in disease symptoms. Races 0 and 1B/C cause yellowing syndrome whereas 1A, 2, 3, 4, 5 and 6 lead to wilting syndrome. Genetics of resistance to two races (1B/C and 6) is yet to be determined, however, for other races resistance is governed either by monogenes or oligogenes. The individual genes of oligogenic resistance mechanism delay onset of disease symptoms, a phenomenon called as late wilting. Slow wilting, i.e., slow development of disease after onset of disease symptoms also occurs in reaction to pathogen; however, its genetics are not known. Mapping of wilt resistance genes in chickpea is difficult because of minimal polymorphism; however, it has been facilitated to great extent by the development of sequence tagged microsatellite site (STMS) markers that have revealed significant interspecific and intraspecific polymorphism. Markers linked to six genes governing resistance to six races (0, 1A, 2, 3, 4 and 5) of the pathogen have been identified and their position on chickpea linkage maps elucidated. These genes lie in two separate clusters on two different chickpea linkage groups. While the gene for resistance to race 0 is situated on LG 5 of Winter et al. (Theoretical and Applied Genetics 101:1155–1163, 2000) those governing resistance to races 1A, 2, 3, 4 and 5 spanned a region of 8.2 cM on LG 2. The cluster of five resistance genes was further subdivided into two sub clusters of 2.8 cM and 2.0 cM, respectively. Map-based cloning can be used to isolate the six genes mapped so far; however, the region containing these genes needs additional markers to facilitate their isolation. Cloning of wilt resistance genes is desirable to study their evolution, mechanisms of resistance and their exploitation in wilt resistance breeding and wilt management.     

The long and winding road leading to the successful introgression of downy mildew resistance into onion

Downy mildew resistance originating from Allium roylei Stearn provides a complete resistance to onions and is based on one, dominant gene. Since A. roylei can successfully be hybridized with onion (A. cepa L.), a breeding scheme aimed at the introgression of this gene was initiated ca. 20 years ago. Several setbacks in this programme were encountered, firstly the identified molecular marker linked to the downy mildew resistance locus became increasingly difficult to use and finally lost its discriminating power and secondly the final step, making homozygous introgression lines (ILs), turned out to be more difficult then was hoped. GISH analysis showed that the chromosomal region harbouring the resistance locus was the only remaining piece of A. roylei in the nuclear background of onion and it also confirmed that this region was located on the distal end of chromosome 3. It was hypothesized that some factor present in the remaining A. roylei region was lethal when homozygously present in an onion genetic background. The identification of an individual with a smaller and more distally located introgression fragment and homozygous ILs in its progeny validated this hypothesis. With the help of these nearly isogenic lines four AFLP^® markers closely linked to the resistance gene were identified, which can be used for marker-aided selection. The introduction of downy mildew resistance caused by Peronospora destructor into onion is a significant step forward in the development of environmentally-friendly onion cultivars.     

Development of Elongated Uppermost Intemode CMS Lines for Hybrid Rice Breeding in India

Development of cytoplasmic male sterile （CMS） lines with elongated uppermost internode （EUI） trait provides a genetic option to eliminate the use of GA3 in hybrid rice. During the past two decades, extensive work has been carried out on the subject that resulted in identification of several mutants with EUI trait for use in developing cytoplasmic/thermo-sensitive genic male-sterile lines with complete panicle exsertion （Okuno and Kawai, 1978; Rutger and Carnahan, 1981; Shen et al., 1987; Yang et al., 2000; 2002; Gangashetti et al., 2004）. Two recessive genes （eui-1 and eui-2） mapped on chromosome 5 and 10 respectively have so far been reported to control the internode elongation in rice （Librojo and Khush, 1986; Yang et al., 2001; Ma et al., 2004）. Considerable progress has been made in China in exploiting ＇eur gene for development of hybrid rice parental lines （Yang et al., 2000; 2002; Zhang et al., 2002）.     

北方粳稻分子育种研究进展

本文介绍了北方粳稻分子育种课题的研究情况和所取得的研究成果。目前主要进行两方面的研究，一方面为特异基因型的鉴定：主要有筛选鉴定出抗旱性材料86份，苗期抗寒性材料40份，后期抗寒性材料629份，抗稻瘟病材料28份，对筛选出的抗旱性材料进行了第二轮聚合杂交，共配组合171个。另一方面是关于遗传与生理基础研究，对穗部性状、粒形、产量和品质以及它们之间的关系进行了深入的研究。另外本文对北方粳稻分子育种材料构建过程中存在的问题进行了综合讨论，并讨论北方粳稻分子育种今后的发展方向。     

RAPD and ISSR molecular markers in <Emphasis Type="Italic">Olea europaea</Emphasis> L.: Genetic variability and molecular cultivar identification

Thirty Portuguese and eight foreign olive (Olea europaea L.) cultivars were screened using Random Amplified Polymorphic DNA (RAPD) and Inter-Simple Sequence Repeat (ISSR) markers. Twenty RAPD primers amplified 301 reproducible bands of which 262 were polymorphic; and 17 ISSR primers amplified 204 bands of which 180 were polymorphic. The percentage of polymorphic bands detected by ISSR and RAPD was similar (88 and 87%, respectively). The genetic variability observed was similar in the Portuguese and foreign olive cultivars. Seven ISSR and 12 RAPD primers were able to distinguish individually all 38 olive cultivars. Twenty specific molecular markers are now available to be converted into Sequence Characterised Amplified Region (SCAR) markers. Relationships among Portuguese and foreign cultivars is discussed.     

Fatty acid composition of various ectomycorrhizal fungi and ectomycorrhizas of Norway spruce

Whole cell fatty acid (WCFA) compositions of three different structures of ectomycorrhizal (ECM) fungi: sporocarps, pure culture mycelia and ectomycorrhizas were analysed to evaluate the potential use of fatty acid profiles as biomarkers for ECM fungi and ectomycorrhiza-associated bacteria. Sporocarps of Amanita muscaria, Amanita rubescens, Lactarius rufus, Lactarius thejogalus, Leccinum scabrum, Paxillus involutus, Russula foetens, Russula rosea, Russula vesca, Suillus grevillei, Tylopilus felleus, Xerocomus badius, Xerocomus subtomentosus, pure cultures of A. muscaria, P. involutus, X. badius, X. subtomentosus, Suillus bovinus Suillus luteus and seven ectomycorrhizal morphotypes of Norway spruce were examined. Our results revealed species-specific composition of fatty acids of fungal sporocarps and pure culture mycelia. Ectomycorrhizal morphotypes distinguished and identified by morphological and molecular methods (PCR-RLFP and sequencing) created specific fatty acid profiles. The dominating fatty acids in pure cultures and sporocarps were 18:2ω6,9, 18:1ω9 and 16:0, whereas ectomycorrhizas also contained plant and bacterial specific fatty acids. Especially, fatty acids specific to Gram-positive bacteria 15:0 anteiso and 17:0 anteiso were present in relatively high amounts and suggested that these bacteria are dominating in the examined Norway spruce mycorrhizosphere. In conclusion, our results show that fatty acid based methods can be useful in studies of ectomycorrhizal fungi, both as a quick method for differentiation of fungal species and also in studies of mycorrhiza-associated microorganisms in the field.     

Inter Simple Sequence Repeat (ISSR) Analysis of Diploid Coffee Species and Cultivated Coffea arabica L. from Tanzania

Inter simple sequence repeat (ISSR) markers were used to evaluate levels of genetic similarity among Coffea arabica L. accessions from Tanzania and to estimate levels of genetic similarities in C. arabica and diploid coffee species. The six ISSR primers used generated a total of 82 fragments and the dissimilarity values ranged from 0.21 to 1. Mean dissimilarity values between provenances (0.56–0.85) were higher than within provenances (0.37–0.68). Cluster analysis based on Nei’s genetic distances showed C. arabica provenances grouping based on geographical origin. Two major clusters were formed that constituted of provenances from Kilimanjaro and Arusha in one sub-cluster; Tanga and Morogoro in the other; the second cluster had Mbeya provenances and diploid species, respectively. The implication is that Mbeya provenances are different from the rest of Tanzanian C. arabica. A principal coordinate analysis (PCA), whose first three coordinates explained 43% of the variation, showed similar groupings as in the cluster analysis. A separate cluster analysis of diploid species showed a distinct separation of the three species used. ISSR data gave results similar to previous findings from random amplified polymorphic DNA(RAPD) analysis. The results also confirm the limited diversity present in cultivated C. arabica in Tanzania