辣椒疫霉漆酶基因Pclac3克隆表达

辣椒疫霉是一种侵染性植物病原,能引起多种茄科及葫芦科植物的病害。利用同源克隆法从辣椒疫霉基因组内克隆了漆酶基因Pclac3,该基因全长1 881个核苷酸,编码626个氨基酸,与已知的真菌漆酶基因具有较高的同源性,并且具备漆酶基因的保守区域。利用PCR将该基因克隆于pPIC9K载体,并转化于毕赤酵母GS115,经过1%甲醇诱导表达获得其异源表达产物。将表达产物进行SDS-PAGE检测,获得分子量大约为90 kDa的特异蛋白。利用ABTS法对Pclac3的表达产物粗酶液进行酶活性分析发现,其活性在第11天时最大,达到45 U/mL。这为研究辣椒疫霉漆酶基因家族及探索卵菌漆酶生物活性及其潜在的应用提供了理论依据。     

重庆地区致病疫霉交配型测定

通过对峙培养在RSA和RTA上对重庆地区主要马铃薯和番茄产区的致病疫霉交配型进行了测定。结果显示,分离自重庆地区8个县(区)的186个致病疫霉菌株均为A1交配型,没有检测到A2交配型。这一结果表明该地区的致病疫霉仍以A1交配型为主。     

Genetic linkage of QTLs for late blight resistance and foliage maturity type in six related potato progenies

A set of test crosses of diploid potatoes was used to identify QTLs for foliage resistance against Phytophthora infestans and QTLs for foliage maturity type, and to assess their genetic relationship. The most important locus for both traits was found on chromosome 5 near marker GP21: the allele of marker GP21 that is associated with resistance to late blight is also associated with late foliage maturity. An additional QTL with a small effect on foliage maturity type was identified on chromosome 3, and additional QTLs for late blight resistance were found on chromosomes 3 and 10. Another QTL was detected on chromosome 7 when resistance was adjusted for the effect of foliage maturity type. The additional QTLs for resistance against P. infestans on chromosomes 3 and 10 seem to be independent of foliage maturity type and are not affected by epistatic effects of the locus on chromosome 5. The effects of the additional QTLs for resistance are small, but early maturing genotypes that necessarily have the allele for susceptibility for late blight on chromosome 5 may benefit from the resistance that is provided by these QTLs on chromosomes 3 and 10.     

Identification of pathogenic races ofPhytophthora fragariae Hickman in Germany

Summary The fungusPhytophthora fragariae Hickman is the causal agent of red stele disease in strawberry (Fragaria × ananassa), this being a major disease in many areas with cool and moist conditions. Success of resistance breeding can be nullified by the appearence of specific races of the fungus capable of overcoming the introduced resistance. In some countries (USA, UK, Japan and Canada) races were identified by using a differential set of strawberry cultivars. The absence of an international standard differential set and the use of different test methods make a comparison of the identified races difficult or even lead to contradictory results for one variety/race combination. The aim of this study was to obtain information about the spectrum of pathogenic races in Germany as a basic contribution to research on resistance breeding against the fungus. The susceptibility of different strawberry cultivars to German isolates ofP. fragariae was evaluated. The inoculation was done by modifying a method described by Milholland et al. (1989). Rootedin vitro plants, four weeks after transferring them to the soil, were used for the investigation. Inoculation was done with a zoospore suspension of defined concentration. Up to now three German isolates, G-1, G-2, and G-3, can be separated by their ability to infect and produce oospores in the roots of the strawberry cultivars ‘Senga Sengana’, either ‘Saladin’, ‘Redgauntlet’ or ‘Climax’ andFragaria chiloensis clone ‘YaquinaB’. These results confirm the existence of pathogenic races ofP. fragariae in Germany and should be taken into consideration for resistance breeding against the fungus.     

Resistance to late blight in potato. A putative gene that suppresses R genes and is elicited by specific isolates

Eleven progenies, ten bi-parental and one selfed, of tetraploid potato were assessed for phenotypic segregation ratios of resistance to susceptible individuals against two isolates of P. infestans, one avirulent to all 11 known potato R genes (avirulent pathotype) and one virulent to eight R genes (virulent pathotype). Assuming Mendelian inheritance of independent R genes, the numbers of individuals segregating for resistance to both races could not be explained sufficiently for all progenies. However, the assumed presence of dominant, specific suppressors of the R gene function that were elicited by specific isolates of the fungus and that segregated in the host independently from R genes explained all frequencies of resistant and susceptible individuals in each of the progenies studied. This revised version was published online in July 2006 with corrections to the Cover Date.     

Multi-trait selection in a diallel crossing scheme of cocoa

Cocoa breeding seeks to increase the yield by area unit. In this context, one must consider the resistance to Phytophthora disease, the most serious one in cocoa culture. Thus, herein two traits were used: yield, i.e. number of healthy pods obtained in 7 years and Phytophthora resistance, estimated by pod rot rate. A combined individual‐family selection is proposed on the basis of the total genetic value in order to spread clonal material. This selection method was applied in a 6 × 6 diallel scheme in Cameroon. The heritability of Phytophthora resistance (near 0.2) was higher than for yield (0.13). A selection index was constructed to obtain good genetic gains on each of the two traits (yield and Phytophthora resistance). To improve prediction of genetic values, a third trait, potential yield, was used. With the chosen weight, the expected genetic gain was about 35% for resistance at 1% level of selection. This selection was applied and a confirmation trial is being established with the selected trees.     

Differentiation of the high molecular weight glutenin subunit D t x 2.1 of Aegilops tauschii indicated by partial sequences of its encoding gene and SSR markers

The devastating late blight pathogen Phytophthora infestans infects foliage as well as tubers of potato. To date, resistance breeding has often focused on foliage blight resistance, but tuber blight resistance is becoming more and more important in cultivated potatoes. In this study, a reliable tuber assay for resistance assessment was developed and foliage and tuber blight resistance (R) was compared in four mapping populations. In the RH4X-103 population, tuber blight resistance inherited independently from foliage blight resistance. Three specific R genes against P. infestans were segregating. The Rpi-abpt and R3a genes function as foliage-specific R genes, whereas the R1 gene acts on both foliage and tuber. In the segregating populations SHRH and RH94-076, tuber and foliage blight resistance correlated significantly, which suggests that resistance in foliage and tuber is conferred by the same gene (could be R3b) and quantitative trait loci (QTL), respectively. In the CE population neither tuber nor foliage resistance was observed.     

Identification of somatic hybrids of dihaploid Solanum tuberosum lines and S. brevidens by species specific RAPD patterns and assessment of disease resistance of the hybrids

Summary Symmetric somatic hybrids were produced by electrofusion of protoplasts of two dihaploid tuber-bearing potato (Solanum tuberosum L.) lines and Solanum brevidens Phil., a diploid non-tuber-bearing wild potato species. A total of 985 plants was obtained. Verification of nuclear hybridity of putative hybrids was based on additive RAPD patterns, general morphological characteristics and chromosome counts. 53 (90%) calli regenerated into plants which were identified as somatic hybrids. Most of the hybrids were aneuploids at the tetraploid (4×) or hexaploid (6×) level. The 20 hybrids tested expressed a high level of resistance to potato virus Y (PVY ^N ) characteristic of the S. brevidens parent. Resistance to late blight (Phytophthora infestans (Mont.) de Bary) varied between hybrids, but was on average better than that of the fusion parents. Resistance of hybrids to bacterial stem rot (Erwinia carotovora subsp. atroseptica (van Hall) Dye) was not superior to that of commercial potato cultivars.     

Linkage and quantitative trait locus mapping of foliage late blight resistance in the wild species Solanum vernei

The global cultivation of potato (Solanum tuberosum) is threatened by epidemics caused by new variants of the late blight pathogen, Phytophthora infestans. New sources of durable late blight resistance are urgently needed and these may be found in wild Solanum species. The diploid wild species, S. vernei, has not previously been subjected to mapping of quantitative trait loci (QTLs) for late blight resistance. Two populations designated HGIHJS and HGG, originating from a cross between a clone of S. vernei and two different S. tuberosum clones were evaluated in field trials for late blight infestation. The relative area under the disease progress curve (RAUDPC) was estimated and used for QTL mapping. A linkage map of S. vernei, comprising 11 linkage groups, nine of which could be assigned to chromosomes, was constructed. Results indicated that the resistance in S. vernei was quantitatively inherited. Significant QTLs for late blight resistance were identified on chromosomes VIII (HGG), VI and IX (HGIHJS). In addition, potential QTLs were detected on chromosomes VII (HGIHJS) and IX (HGG). A putative and a significant QTL for tuber yield were found on chromosomes VI and VII in HGG, but no linkage between yield and resistance was indicated. The QTL for late blight resistance, which mapped to chromosome IX, could be useful for late blight resistance breeding as it was located close to the microsatellite marker STM1051 in both populations.     

Gamma Ray-Induced Field Tolerance to Phytophthora Blight in Sesame

Due to the non-availability of sources of resistance in Sri Lanka to Phytopbthora blight in sesame (Sesamum indicum) caused by Phytopbthora nicotianae var. parasitica, a mutation breeding programme was initiated. Seeds of three genotypes were subjected to six doses of gamma rays from 100 Gy to 750 Gy from a ⁶⁰Co source. Seeds of M₂ bulks, sampled by variety and treatment from the first five capsules formed on M₁ plants, were grown in a field with a history of repeated incidence of the disease. The best 21 lines, having the highest survival and seed production, identified in the screening of M₃ and M₄ progeny rows of selected M₂ single plants were then tested in a replicated field trial with the recommended cultivar ‘MI 3’ as a control. The plant survival in selected lines averaged 43.3 percent as against 7.2 percent in the recommended variety. Eight selections recorded significantly higher seed yield than ‘MI 3’ at P < 0.01 and another three at P < 0.05. Gamma ray treatments of 450 Gy and 600 Gy produced more lines tolerant to the disease than the other doses used.