New genes for disease resistances through somatic hybridization

Somatic hybridization, a process of combining protoplasts from different plants, can provide new sources of disease resistances for plants. In the case of wild and cultivatedSolanum species, the hybrids express resistances from each partner in the fusion and can often be crossed with cultivars to improve agronomic characteristics of the tubers. Restriction fragment length polymorphism (RFLP) analyses can provide a means for determining that the plants being investigated are actually hybrids as well as a means for following the introgression of DNA into progeny lines. These points are addressed in this paper with specific reference to somatic hybrids betweenSolanum brevidens and potato.     

The effects of agricultural fields and human settlements on the use of rivers by wildlife in the mid-Zambezi valley,Zimbabwe

After the eradication of the Tse-Tse fly in the Mid-Zambezi valley, human settlements and fields extended mainly along the main rivers. In order to investigate the consequences of this human development on wildlife diversity we monitored three rivers of the Mid-Zambezi valley in Zimbabwe: Angwa, Manyame and Kadzi. The rivers were divided in segments of 200 m which were checked for spoors in order to assess the number of species and the number of individuals that used the segments. Human settlements were also recorded. We used a GIS to define the spatial characteristics of the fields present along the rivers, and related them to the distribution and abundance of wild species spoors in the river beds and banks. Our results show that the number of species in one segment of the river decreased with the increasing size of the field area bordering the segment. For all the major ungulate species, the numbers of individuals recorded per segment decreased with increasing field area. A similar trend was observed for small and medium-sized carnivores, though they were in lower numbers when present. Our analyses thus confirm that the extension of human agriculture in wildlife areas has an impact on most wild species, but we also define some threshold value of field size above which there seem to be an acceleration of the decrease in wildlife density and diversity: 3.2 ha for medium and small herbivores and carnivores; only the elephant seem to tolerate larger field area with a threshold value of 32 ha.This revised version was published online in May 2005 with corrections to the Cover Date.     

Effects of land ownership and landscape-level factors on rare-species richness in natural areas of southern Ontario,Canada

Surprisingly few studies have considered the extent to which the nature of the ownership of land is associated with differences in biodiversity. We analysed ownership and other landscape-level effects on rare-species richness for both globally- and regionally-rare biota (including birds, herpetofauna, butterflies, mammals, and plants) in 289 designated natural areas (NAs) in southern Ontario, Canada. Information about each NA −including area, number of plant communities, ownership status and details of species diversity were collected from published sources. Length of perimeter of NA, relative isolation, and an estimate of fragmentation were measured using image analysis and GIS techniques. NAs were in general relatively small, with mean area of 158 ha (median 85 ha, range from 0.9 to 1278 ha) for private NAs; public NAs had mean area of 132 ha (median 16 ha, range from 0.1 to 1481 ha). Mean number of plant communities was 4.6 (median 4, range 1- 13) at private NAs and 3.8 (median 3, range 1-16) at public NAs. Our results show that, of several landscape-level factors, area had the greatest effects on rare-species richness and other biotic indices. Effects of area were followed by effects of plant community diversity, however this was itself significantly affected by area and the extent of perimeter of the NA. Both these factors were followed by effects of ownership of the NA and by effects of isolation of the NA (represented by minimum distance to nearest NA and by number of NAs in 10 km radius). Other landscape- level factors did not appear to have overall significant effects. Variation in area accounted for 0.1% to 29% of variation in number of rare species, with lower values for globally-rare, than for regionally-rare taxa. For all biotic groups, public ownership of NAs was associated with significantly greater rare-species richness compared to private ownership, even after other factors such as area were controlled. For all globally-rare biota except butterflies, area of NA had greater effects on rare-species richness than did ownership. Richness of regionally- rare birds was more affected by plant community diversity than by area of NA. Number of recorded plant communities accounted from 2.1% of variation in number of globally-rare plant species to as high as 31% of variation in regionally-rare butterflies. The diversity of plant communities was itself influenced by total site area (accounting for 45% of variation), extent of elongation of the NA, and both external- and interior- edge perimeters. Public NAs had greatest numbers of rare biota and so should be a significant focus for conservation programs. Smaller, privately-owned patches of natural area dominate (by number and area) in this densely populated region and their significance should not be overlooked. This revised version was published online in July 2006 with corrections to the Cover Date.     

Detection of the Defoliating Pathotype of Verticillium dahliae in Infected Olive Plants by Nested PCR

Spread of Verticillium wilt into newly established olive orchards in Andalucía, southern Spain, has caused concern in the olive industry in the region. This spread may result from use of Verticillium dahliae-infected planting material, which can extend distribution of the highly virulent, defoliating (D) pathotype of V. dahliae to new areas. In this study, a molecular diagnostic method for the early in planta detection of D V. dahliae was developed, aimed especially at nursery-produced olive plants. For this purpose, new primers for nested PCR were designed by sequencing a 992-bp RAPD marker of the D pathotype. The use of the specific primers and different nested-PCR protocols allowed the detection of V. dahliae pathotype D DNA in infected root and stem tissues of young olive plants. Detection of the pathogen was effective from the very earliest moments following inoculation of olive plants with a V. dahliae pathotype D conidia suspension as well as in inoculated, though symptomless, plants.     

Observations of Phytophthora spp. in Water Recirculation Systems in Commercial Hardy Ornamental Nursery Stock

Samples of water and sediment were taken from drains, reservoirs and wells from four commercial hardy ornamental nurseries with water recirculation systems. The samples were taken on seven different dates throughout a single year from August 1994 to July 1995. The samples were screened for Phytophthora species using five different methods: direct plating, three bait tests (using lupin seedlings, apples and Rhododendron leaves) and a DAS-ELISA (double-antibody sandwich enzyme-linked immunosorbent-assay) with two antisera. In the nurseries with old water recirculation systems, Phytophthora species were detected in the drains and in the reservoirs. In the nursery with a new recirculation system, the pathogens were only present in the drains. None of the water samples from wells in any of the nurseries were contaminated. Phytophthora species were present in the water as well as in the sediment samples from drains and reservoirs. They were detected in the water recirculation systems irrespective of the season. The number of isolates increased about sevenfold between late summer and spring. At least 12 different Phytophthora species were identified: some isolates were previously unrecorded species. The epidemiology of the pathogens in outdoor water recirculation systems as well as the importance of the results for commercial nurseries is discussed.     

Induction of an antioxidant enzyme system and other oxidative stress markers associated with compatible and incompatible interactions between chickpea (Cicer arietinum L.) and Fusarium oxysporum f. sp.ciceris

To ascertain if active oxygen species play a role in fusarium wilt of chickpea caused by Fusarium oxysporum f. sp. ciceris, the degree of lipid peroxidation (malondialdehyde formation) and the activity levels of diamine oxidase (DAO), an apoplastic H₂O₂-forming oxidase, and several antioxidant enzymes, namely ascorbate peroxidase (APX), catalase (CAT), glutathione reductase (GR), guaiacol-dependent peroxidase (GPX) and superoxide dismutase (SOD), were determined spectrophotometrically in roots and stems of ‘WR315’ (resistant) and ‘JG62’ (susceptible) chickpea cultivars inoculated with the highly virulent race 5 of the pathogen. Moreover, APX, CAT, GPX and SOD were also analysed in roots and stems by gel electrophoresis and activity staining; and the protein levels of APX and SOD in roots were determined by Western blotting. In roots, infection by the pathogen increased lipid peroxidation and CAT and SOD activities, although such responses occurred earlier in the incompatible compared with the compatible interactions. APX, GPX and GR activities were also increased in infected roots, but only in the compatible interaction. In stems, infection by the pathogen increased lipid peroxidation and APX, CAT, SOD and GPX activities only in the compatible interaction, and DAO activity only in the incompatible one. In general, electrophoregrams agreed with the activity levels determined spectrophotometrically and did not reveal any differences in isoenzyme patterns between cultivars or between infected and non-infected plants. Further, Western blots revealed an increase in the root protein levels of APX in the compatible interaction and in those of SOD in both compatible and incompatible interactions. In conclusion, whereas enhanced DAO activity in stems, and earlier increases in lipid peroxidation and CAT and SOD activities in roots, can be associated with resistance to fusarium wilt in chickpea, the induction of the latter three parameters in roots and stems along with that of APX, GR (only in roots) and GPX (only in stems) activities are rather more associated with the establishment of the compatible interaction.     

ITS—RFLP分析进行昆虫病原线虫斯氏属和异小杆属的分类鉴定

本研究通过PCR扩增和六种限制性内切酶（AluⅠ，HinfⅠ，MboⅠ，RsaⅠ，HaeⅢ和PvuⅡ）酶切，对国内害虫防治上常用的几种昆虫病原线虫，包括斯氏属S.car-pocapsae,S.feltiae和S.glaseri以及异小杆属H.bacteriophora,H.zealandica,H.indicus和H.megidis等8个品系rDNA-ITS进行分析。建立起可以区分各线虫种的标准RFLP图谱。该方法快速简便，稳定可靠，需要的样品量少。可以用于新鲜的，或冻存的样品，甚至分析单条的线虫，不仅可进行昆虫病原线虫的快速分类鉴定。而且进一步可以应用于线虫田间释放的辅助监测。实际田间感染率的测定和线虫毒力的比较。     

山西菜田捕食性食蚜蝇优势种及其对菜蚜的控制效果

对山西菜田捕食性食蚜蝇优势种及对菜蚜的控制效果进行了研究，菜田常见捕食性食蚜蝇有黑带食蚜蝇，大灰食蚜蝇，斜斑鼓额食蚜蝇，野食蚜蝇，四条小食蚜蝇，短翅细腹食蚜蝇6种，以黑带食蚜蝇种群数量最大，平均占全年发生总量的36.3%，通过对食蚜蝇产卵量，卵化率，幼虫期捕食量和羽化率测试，筛选出的黑带食蚜蝇，大灰食蚜蝇，斜斑鼓额食蚜蝇幼虫对菜蚜均有较强的捕食能力，其中黑带食蚜蝇为山西菜田菜蚜捕食性食蚜蝇优势种，用黑带食蚜蝇三龄幼虫对菜蚜进行防治试验，结果以1：180防效最佳，72h后控制效果达93.5%。