Inhibition of tobacco NADH-hydroxypyruvate reductase by expression of a heterologous antisense RNA derived from a cucumber cDNA: Implications for the mechanism of action of antisense RNAs

Tobacco plants were genetically transformed to generate antisense RNA from a gene construct comprised of a full-length cucumber NADH-dependent hydroxypyruvate reductase (HPR) cDNA placed in reverse orientation between the cauliflower mosaic virus 35S promoter and a nopaline synthase termination/polyadenylation signal sequence. In vivo accumulation of antisense HPR RNA within eight independent transgenic tobacco plants resulted in reductions of up to 50% in both native HPR activity and protein accumulation relative to untransformed tobacco plants (mean transgenote HPR activity=67% wild type, mean transgenote HPR protein=63% wild type). However, in contrast to previous reports describing antisense RNA effects in plants, production of the heterologous HPR antisense RNA did not systematically reduce levels of native tobacco HPR mRNA (mean transgenote HPR mRNA level=135% wild type). Simple regression comparison of the steady-state levels of tobacco HPR mRNA to those of HPR antisense RNA showed a weak positive correlation (r value of 0.548, n=9 ; n is wild type control plus eight independent transformants; significant at 85% confidence level), supporting the conclusion that native mRNA levels were not reduced within antisense plants. Although all transgenic antisense plants examined displayed an apparent reduction in both tobacco HPR protein and enzyme activity, there is no clear correlation between HPR activity and the amount of either sense (r=0.267, n=9) or antisense RNA (r=0.175, n=9). This compares to a weak positive correlation between HPR mRNA levels and the amount of HPR activity observed in wild-type SRI tobacco plants (r=0.603, n=5). The results suggest that in vivo production of this heterologous HPR antisense RNA is inhibitory at the level of HPR-specific translation and produces its effect in a manner not dependent upon, nor resulting in, a reduction in steady-state native HPR mRNA levels. In this context, the observed antisense effect appears to differ mechanistically from most antisense systems described to date.     

Evidence for nuclear internalization of exogenous DNA into mammalian sperm cells

Mature sperm cells have the spontaneous capacity to take up exogenous DNA. Such DNA specifically interacts with the subacrosomal segment of the sperm head corresponding to the nuclear area. Part of the sperm-bound foreign DNA is further internalized into nuclei. Using end-labelled plasmid DNA we have found that 15–22% of the total sperm bound DNA is associated with nuclei as determined on isolated nuclei. On the basis of autoradiographic analysis, nuclear permeability to exogenous DNA seems to be a wide phenomenon involving the majority of the sperm nuclei. In fact, the foreign DNA, incubated with sperm cells for different lengths of time, is found in 45% (10 min) to 65% (2 hr) of the sperm nuclei. Ultrastructural autoradiography on thin sections of mammalian spermatozoa, preincubated with end-labelled plasmid DNA, shows that the exogenous DNA is internalized into the nucleus. This conclusion is further supported by ultrastructural autoradiographic analysis on thin sections of nuclei isolated from spermatozoa preincubated with end-labelled DNA. © 1993 Wiley-Liss, Inc.     

中国优质水果资源的分布与适宜生态环境

根据农业部在80年代两次组织评选出的全国189个优质水果产地的生态环境资料,用微型电子计算机系统建立数据库,统计分析柑桔、苹果和梨优质产品的构成比例、产区分布地域及其适宜的环境指标和主栽品种的生态适应性,为果树良种区域化栽培与选育提供依据。     

Isolation of RNA from apple skin

A method has been developed for the isolation of RNA from apple skin. The method involves an adaptation of the Manning (1991) method and includes a high-salt extraction step and a final purification step through a CsCl cushion. The RNA isolated was of high quality and produced good hybridization signals in northern blot analyses.     

Oviposition preference of Anthocoris nemorum and A. nemoralis for apple and pear

Anthocoris nemorum L. and Anthocoris nemoralis Fabricius (Heteroptera: Anthocoridae) are important predators of insect pests in pome fruit. Females insert their eggs in leaf tissue. The females’ choice of oviposition site is important for the subsequent distribution of nymphs on host plants. Oviposition preference for apple and pear leaves was tested in the laboratory in four experiments (experiments 1–4). In three experiments it was tested whether simulated insect damage to leaves (experiments 5 and 6) or the presence of prey (experiment 7) influenced oviposition preference. The effect of the presence of prey was only tested for A. nemorum on apple leaves. There was a highly significant anthocorid species × plant interaction for the number of eggs laid on apple and pear leaves. Anthocoris nemorum laid more eggs on apple than on pear leaves, while A. nemoralis preferred pear. Anthocoris nemorum's preference for apple increased over the 6‐week period in which experiments 1–4 were performed, from 66% to 91% eggs laid on apple leaves. No change over time in preference was found for A. nemoralis. Across experiments 1–4, the majority of A. nemorum eggs were laid near leaf margins, whereas eggs of A. nemoralis were more commonly found in the leaf centre, 5 mm or more from the margin, with a highly significant leaf region × species interaction. There was no significant difference in preference for leaf side between A. nemorum and A. nemoralis, but there was a highly significant plant × leaf side × experiment interaction. Thus, more eggs were laid on the ventral than on the dorsal side of pear leaves in experiment 4, while significantly more eggs were laid on the dorsal side of apple leaves in experiments 3 and 4. Choice tests between damaged and healthy leaves showed that A. nemorum laid significantly more eggs on the damaged leaves, while A. nemoralis preferred healthy leaves. Anthocoris nemorum showed a near‐significant preference for ovipositing on leaves with eggs of Operophtera brumata (Lepidoptera: Geometridae). The oviposition preferences found correspond to the natural distribution of these predators in apple and pear orchards. The preference of A. nemorum for leaf margins, and of A. nemoralis for the leaf centre as an oviposition site, supports earlier observations. A preference for leaf side for oviposition site has not been reported earlier. Preference for damaged leaves could help A. nemorum to locate prey in a field situation.     

Summer Epidemics of Apple Scab: The Relationship between Measurements and their Implications for the Development of Predictive Models and Threshold Levels under Different Disease Control Regimes

A 2‐year study on epidemic progress of apple scab was conducted at Randwijk, the Netherlands, in 1998 and 1999. The summer epidemic caused by conidia was studied instead of the well‐described spring season epidemic originating from ascospores. The aim was to investigate relationships between disease measurements, i.e. disease incidence and severity measures of apple scab, and their implications for the development of predictive models and threshold levels. The study characterized good relationships between the measurements on cultivar Jonagold using regression analyses in three disease control regimes (untreated, organic and integrated). For fruit quality prediction, the relationship between fruit incidence (I_f) and leaf incidence (I_l) in the organic control regime was given by I_f = 1.966 + 0.402 × (I_l) (R² = 0.92). As a result of low level of disease in the integrated control regime, shoot incidence (I_s), with higher values than leaf incidence, was better suited for prediction. The relationship was given by I_f = ?0.162 + 0.028 × (I_s) (R² = 0.91). For the integrated control regime, disease threshold levels were constructed for timing of the final fungicide application. If an apple grower wants to keep fruit infection under 1% incidence (harvest scab threshold), the timing of the final fungicide application (action threshold) should correspond to 4% shoot scab incidence at the beginning of August. The results are compared with similar studies and their biological interpretation is discussed.     

Epidemiology of apple proliferation (AP) in northwestern Italy: evaluation of the frequency of AP-positive psyllids in naturally infected populations of Cacopsylla melanoneura (Homoptera: Psyllidae)

Adults of Cacopsylla melanoneura, vector of the apple proliferation (AP) phytoplasma, were collected every 2 weeks from January until May in 2000 and 2001 by the beating tray method in eight apple orchards of the Aosta Valley (northwestern Italy). Total DNA was extracted from batches of five insects and amplified with the universal phytoplasma primers P1/P7 in direct PCR. A nested PCR assay was then performed on P1/P7 amplicons using the primers fO1/rO1, specific for the AP‐ phytoplasma group. The digestion of fO1/rO1 amplicons with Ssp I restriction endonuclease confirmed that C. melanoneura adults harboured the AP phytoplasma. The data obtained with PCR were used to estimate the proportion of AP‐positive insects in over wintered and offspring adults. Percentages of AP‐positive insects of 3.6% and 0.8% were estimated in 2000 among over wintered and offspring psyllids respectively. In 2001 only the over wintered insects were found infected, with an estimated proportion of 2.8%. The seasonal abundance of the vector was measured using yellow sticky traps. C. melanoneura was always present at a low population level, and the highest density was recorded from mid‐February until mid‐March in both years. The results show that the overwintered population is higher and spends a longer period in apple orchards, suggesting the crucial role of the overwintered adults in vectoring AP.     

复制型HBV转基因小鼠的遗传与繁育研究

本文采用类似系祖建系的方法,对复制型ＨＢＶ转基因小鼠模型的１７、２１、２５三个系列进行了繁育,通过ＰＣＲ和Ｓｏｕｔｈｅｒｎ分子杂交的方法,检测三个系列小鼠尾组织和血清的ＤＮＡ样品,以确定ＨＢＶ基因的整合和复制情况。结果表明,ＨＢＶ基因已稳定地遗传至第五代（Ｆ５）,且各世代小鼠血清中都存在ＨＢＶＤＮＡ,此外,整合阳性小鼠的血清中能测出ＨＢＶＤＮＡ的比率很高,Ｆ１代为９４．４４％（１０２／１０８）,Ｆ２、Ｆ３代为９５．３１％（６１／６４）,故在繁育中可以通过测血清中的ＨＢＶＤＮＡ来确定小鼠是否整合。