抗阿特拉津转基因大豆植株后代的遗传分析

本试验用阿特拉津溶液涂抹、荧光诱导动力学检测、分子杂交等方法对抗阿特拉津转基因大豆植株的后代进行了鉴定,在第二代及第三代中检测到了抗性基因的存在,表明从龙葵中得到的此抗阿特拉津 psbA 基因不仅能导人大豆叶绿体基因组中获得表达,而且可以遗传到后代。     

Regulation of expression of a wound-inducible tomato inhibitor I gene in transgenic nightshade plants

A wound-inducible proteinase Inhibitor I gene from tomato containing 725 bp of the 5 region and 2.5 kbp of the 3 region was stably incorporated into the genome of black nightshade plants (Solanum nigrum) using an Agrobacterium Ti plasmid-derived vector. Transgenic nightshade plants were selected that expressed the tomato Inhibitor I protein in leaf tissue. The leaves of the plants contained constitutive levels of the inhibitor protein of up to 60 g/g tissue. These levels increased by a factor of about two in response to severe wounding. Only leaves and petioles exhibited the presence of the inhibitor, indicating that the gene exhibited the same tissue specificity of expression found in situ in wounded tomato leaves. Inhibitor I was extracted from leaves of wounded transformed nightshade plants and was partially purified by affinity chromatography on a chymotrypsin-Sepharose column. The affinity-purified protein was identical to the native tomato Inhibitor I in its immunological reactivity and in its inhibitory activity against chymotrypsin. The protein exhibited the same M _r of 8 kDa as the native tomato Inhibitor I and its N-terminal amino acid sequence was identical to that of the native tomato inhibitor I, indicating that the protein was properly processed in nightshade plants. These expriments are the first report of the expression of a member of the wound-inducible tomato Inhibitor I gene family in transgenic plants. The results demonstrate that the gene contains elements that can be regulated in a wound-inducible, tissuespecific manner in nightshade plants.     

Persistence of the flowerbud weevil Anthonomus santacruzi in optimal versus marginal areas: implications for the biological control of the invasive tree Solanum mauritianum in South Africa

Anthonomus santacruzi Hustache (Coleoptera: Curculionidae) was released in South Africa to offset the extensive reproductive output of the invasive tree Solanum mauritianum Scopoli (Solanaceae). Widespread establishment has occurred predominantly in the coastal areas of KwaZulu-Natal province, with limited success in higher-altitude inland areas. Irrespective of location, populations exhibit peaks in the austral autumn and decline during winter. In this study, we evaluated the persistence of A. santacruzi populations in climatically-optimal coastal areas versus climatically-marginal inland areas. The weevil’s pre- and post-winter abundance was surveyed at six coastal and six inland sites during 2018, and compared between areas and seasons. The 2018 pre-winter data were also compared to 2016 pre-winter data collected at the same sites. During 2018, pre- and post-winter numbers were six times and 22 times higher, respectively, at optimal sites than at marginal sites, with substantial winter declines at all sites. Post-winter weevil numbers at optimal sites were significantly higher than pre-winter numbers at marginal sites. Pre-winter numbers at optimal sites were not significantly different between 2016 and 2018, but at marginal sites were significantly lower in 2018. Inflorescences of S. mauritianum at marginal sites contained significantly more floral material and fruit than those at optimal sites, during both seasons in 2018. Significant negative correlations between A. santacruzi numbers and floral/fruit production suggest some impact on the reproductive output of S. mauritianum. Since A. santacruzi populations are barely persisting in marginal areas, releases in other South African provinces should target locations that are below 300?m in altitude.     

Biology and host range of Platyphora semiviridis, a leaf beetle evaluated as a potential biological control agent for Solanum mauritianum in South Africa

The South American tree Solanum mauritianum is a major environmental weed in the high-rainfall regions of South Africa and has been targeted for biological control. Potential agents included five species of the genus Platyphora, which were imported from South America in 1994. Platyphora species associated with Solanaceae reputedly have very specific habitat requirements and host plant preferences in the field. Despite this, host-specificity tests on one species, Platyphora semiviridis, revealed a broad physiological host range. Although laboratory tests showed that P. semiviridis is confined to Solanum species and cannot survive on solanaceous crops outside that genus, it developed on potato and cultivated eggplant (aubergine) as well as on 10 native South African Solanum species. With few exceptions, there were no consistent differences in survival and duration of development on these compared with S. mauritianum. Furthermore, at least six of these non-target species, including potato and eggplant, supported breeding colonies of the beetles in cages. During choice tests in both small and larger cages, P. semiviridis avoided potato but did not consistently discriminate between S. mauritianum, eggplant and six native solanums for larviposition. Despite these findings, P. semiviridis has never been recorded on either potato or eggplant in South America, where it was only observed to feed on S. mauritianum. Although there are several reasons why P. semiviridis is unlikely to attack non-target Solanum species in the field, it will not be released in South Africa because there are other imported agents which have displayed narrower physiological host ranges and which may be more effective.     

Patterns of clonal diversity in three species of sub-arctic willow (Salix lanata, Salix lapponum and Salix herbacea)

Sub-arctic willow scrub is an endangered habitat in Britain, and typically occurs on steep crags inaccessible to grazing animals. These willows can reproduce both sexually and asexually, although the relative importance of each is unknown. Knowledge of reproductive mode is important for the design of grazing management and restoration programmes. Accordingly, clonality was assessed in the largest stand of sub-arctic willow scrub in the UK, focusing on Salix lanata and S. lapponum. Little evidence of clonal growth was detected; most individuals possessed distinct multi-locus genotypes. Thus despite the capacity for vegetative reproduction, and seedlings being rarely observed, sexual reproduction is the predominant means of perpetuation and dispersal at this site. We also examined clonal growth in a common willow species (Salix herbacea) that occupies a different habitat type (exposed mountain tops and ridges). Multiple individuals shared identical genotypes up to 7 m apart, suggesting an important role for clonal growth in local patch formation in this species.     

The use of mutagens to increase the efficiency of the androgenic progeny production in <Emphasis Type="Italic">Solanum nigrum</Emphasis>

Pollen embryogenesis was successfully induced in Solanum nigrum L. (2n=6×=72). Stimulation of androgenesis expressed as the frequency of androgenic responsive anthers was observed after 10 and 20 mM ethyl methanesulphonate (EMS), 10 and 20 mM sodium azide (NaN₃) and 0.2 mM N-nitroso-N-methylurea (MNU) treatment applied on seeds for 24 h. The frequency of androgenesis on the medium with sucrose was higher than on the medium with maltose. Androgenic regenerants originated also in the anthers collected from donor plants where survival after mutagenic treatment was lower than 50 %. Green haploid (3x), aneuploid (to 8x) and dihaploid (6x) plants were obtained. The high frequency of aneuploids among androgenic plants is explained by cell division irregularities in microsporial calli.     

A proteinase inhibitor II of Solanum americanum is expressed in phloem

Although proteinase inhibitor proteins are known to confer insect resistance in transgenic plants, their endogenous roles remain undefined. Here, we describe the expression of a proteinase inhibitor II (PIN2) protein from Solanum americanum in phloem of stems, roots and leaves suggesting a novel endogenous role for PIN2 in phloem. The phloem consists of parenchyma cells, sieve elements (SE), and companion cells (CC) which are in close association with SE. We isolated two cDNAs encoding PIN2, SaPIN2a and SaPIN2b, from a S. americanum cDNA library using a tomato PIN2 cDNA as hybridization probe. SaPIN2a shows 73.6% identity to SaPIN2b. Southern blot analysis confirmed that two genes occur in S. americanum. Northern blot analysis showed that both are wound-inducible and are expressed in flowers. Unlike SaPIN2b and other previously characterized plant PIN2 proteins, SaPIN2a is abundantly expressed in stems. In situ hybridization studies on stem sections showed that SaPIN2a mRNA is expressed in CC and some SE, likely the immature developing SE, of external and internal phloem. Western blot analysis using SaPIN2a-specific antibodies showed SaPIN2a accumulation in stems, leaf midribs and fruits. Immunohistochemical localization, using these antibodies, revealed SaPIN2a expression in external and internal phloem of stem. Immunoelectron microscopy of stem, root and leaf sections further localized SaPIN2a to the CC and predominantly to the SE, particularly the parietal cytoplasm adjacent to the cell wall, the lumen and the sieve-area pores. These results suggest that, other than a possible role in plant defense, SaPIN2a could be involved in regulating proteolysis in the SE.     

Bioherbicidal enhancement and host range expansion of a mycoherbicidal fungus via formulation approaches

Eastern black nightshade (Solanum ptycanthum; EBN) is a problematic weed partly due to its tolerance or resistance to certain herbicides. We examined the effects of an invert emulsion (IE) on the host range and weed control efficacy of the fungus Colletotrichum coccodes (NRRL strain 15547) for biocontrol of EBN. Greenhouse tests demonstrated that several other solanaceous weeds were also infected and killed, and field tests revealed >90% EBN control and dry weight reduction in plants treated with the fungus-IE formulation. These results demonstrate that this IE formulation can promote the efficacy of this bioherbicidal pathogen.     

Effects of potnetial trap crops and planting date on soil infestation with potato cyst nematodes and root-knot nematodes

In 1997 and 1998 the stimulation of hatch of potato cyst nematodes (PCN) by a trap crop was studied at various times during the growing season in a container and a field experiment. Solanum nigrum‘90‐4750‐188’was used as the trap crop in both experiments and was sown on 1 May, 16 June or 1 August in two successive years on different plots. Neither experiment revealed much seasonal variation in hatchability of PCN juveniles under a trap crop. In the container experiment, the hatch of the Globodera pallida Pa3 population was equally and strongly stimulated (89%) at all sowing dates in both years, except for the 1 August sowing in 1998 (when the hatch was 77% under extremely wet soil conditions). In the control treatment with non‐hosts (flax followed by barley) the total spontaneous hatch was 50% over 2 yr. In the field experiment, the hatch of PCN, averaged over the four populations, was also equally stimulated (71%) at all sowing dates in both years. In the control treatment with non‐hosts (flax‐barley) the total spontaneous hatch was 36% over 2 yr. Total hatch under the trap crop over 2 yr varied between the four PCN populations from 63% to 80%. In 1998 and 1999, control of potato cyst nematodes (PCN) by the potential trap crops Solanum sisymbriifolium and S. nigrum‘90‐4750‐188’was studied in the field. Potato was also included as a trap crop. In the 1998 experiment, potato, S. sisymbriifolium and S. nigrum strongly stimulated the hatch of PCN compared with the non‐host white mustard (Sinapis alba). Roots of potato and white mustard were mainly found in the top 10 cm of soil, whereas roots of S. sisymbriifolium and S. nigrum were also abundant at depths of 10–20 cm and 20–30 cm. In the 1999 experiment, soil infestation with PCN decreased markedly with potato and S. sisymbriifolium as trap crops. In plots moderately to severely infested with 2‐yr old cysts (2–29 juveniles ml^?1 air dried soil), potato reduced soil infestation by 87% and S. sisymbriifolium by 77%. In plots moderately to severely infested with 1‐yr old cysts the reductions were 74% and 60%, respectively. The reduction was least on plots very severely infested with PCN (110–242 juveniles ml^?1 soil): 69% and 52% for potato and S. sisymbriifolium, respectively. Soil infestations of plots that were initially slightly to severely infested with the root‐knot nematode Meloidogyne hapla were greatly reduced under fallow and S. sisymbriifolium but increased under potato. From these and previous experiments it was concluded that, for several reasons, S. sisymbriifolium is a promising trap crop.     

Assessing the risks of releasing a sap-sucking lace bug, <Emphasis Type="Italic">Gargaphia decoris</Emphasis>, against the invasive tree <Emphasis Type="Italic">Solanum mauritianum</Emphasis> in New Zealand

The South American tree Solanum mauritianum Scopoli (Solanaceae), a major environmental weed in South Africa and New Zealand, has been targeted for biological control, with releases of agents restricted to South Africa. The leaf-sucking lace bug, Gargaphia decoris Drake (Tingidae), so far the only agent released, has become established in South Africa with recent reports of severe damage at a few field sites. To evaluate the insect’s suitability for release in New Zealand, host-specificity testing was carried out in South Africa in laboratory and open-field trials, with selected cultivated and native species of Solanum from New Zealand. No-choice tests confirmed the results of earlier trials that none of the three native New Zealand Solanum species are acceptable as hosts. Although the cultivated Solanum muricatum Aiton and S. quitoense Lam. also proved unacceptable as hosts, some cultivars of S. melongena L. (eggplant) supported feeding, development and oviposition in the no-choice tests. Although eggplant was routinely accepted under laboratory no-choice conditions in this and previous studies, observations in the native and introduced range of G. decoris, open-field trials and risk assessment based on multiple measures of insect performance indicate that the insect has a host range restricted to S. mauritianum. These results strongly support the proposed release of G. decoris in New Zealand because risks to non-target native and cultivated Solanum species appear to be negligible. An application for permission to release G. decoris in New Zealand will be submitted to the regulatory authority. Handling editor: John Scott.