Hu Gang, St Leger Raymond J
Department of Entomology, University of Maryland, College Park, Maryland 20742.
Appl Environ Microbiol. 2002 Dec;68(12):6383-7. doi: 10.1128/AEM.68.12.6383-6387.2002.
In the summer of 2000, we released genetically altered insect-pathogenic fungi onto a plot of cabbages at a field site on the Upper Marlboro Research Station, Md. The transformed derivatives of Metarhizium anisopliae ARSEF 1080, designated GPMa and GMa, carried the Aequorea victoria green fluorescent protein (gfp) gene alone (GMa) or with additional protease genes (Pr1) (GPMa). The study (i) confirmed the utility of gfp for monitoring pathogen strains in field populations over time, (ii) demonstrated little dissemination of transgenic strains and produced no evidence of transmission by nontarget insects, (iii) found that recombinant fungi were genetically stable over 1 year under field conditions, and (iv) determined that deployment of the transgenic strains did not depress the culturable indigenous fungal microflora. The major point of the study was to monitor the fate (survivorship) of transformants under field conditions. In nonrhizosphere soil, the amount of GMa decreased from 10(5) propagules/g at depths of 0 to 2 cm to 10(3) propagules/g after several months. However, the densities of GMa remained at 10(5) propagules/g in the inner rhizosphere, demonstrating that rhizospheric soils are a potential reservoir for M. anisopliae. These results place a sharp focus on the biology of the soil/root interphase as a site where plants, insects, and pathogens interact to determine fungal biocontrol efficacy, cycling, and survival. However, the rhizospheric effect was less marked for GPMa, and overall it showed reduced persistence in soils than did GMa.
2000年夏天,我们将经过基因改造的昆虫病原真菌释放到马里兰州上马尔伯勒研究站一块地里的卷心菜上。金龟子绿僵菌ARSEF 1080的转化衍生物,命名为GPMa和GMa,单独携带维多利亚多管发光水母绿色荧光蛋白(gfp)基因(GMa)或带有额外的蛋白酶基因(Pr1)(GPMa)。该研究(i)证实了gfp在长期监测田间种群中病原体菌株方面的效用,(ii)表明转基因菌株几乎没有传播,也没有发现非靶标昆虫传播的证据,(iii)发现在田间条件下重组真菌在1年多的时间里基因稳定,(iv)确定转基因菌株的投放并未抑制可培养的本地真菌微生物群落。该研究的重点是监测田间条件下转化体的命运(存活率)。在非根际土壤中,GMa的数量从0至2厘米深度处的10⁵个繁殖体/克在几个月后降至10³个繁殖体/克。然而,GMa在根内际的密度保持在10⁵个繁殖体/克,这表明根际土壤是金龟子绿僵菌的一个潜在储存库。这些结果突出了土壤/根系界面的生物学特性,作为植物、昆虫和病原体相互作用以确定真菌生物防治效果、循环和存活的场所。然而,GPMa的根际效应不太明显,总体而言,它在土壤中的持久性比GMa低。