杜仲内生真菌DZJ07的GFP标记及在小麦植株中的定殖

GFP-Tagging and Colonization Observation of the Endophytic Fungus DZJ07 from Eucommia ulmoides

杜仲内生真菌DZJ07对小麦纹枯病菌有较好的防治作用,为了阐明其在小麦植株中的生态适应性,通过农杆菌介导转化法将绿色荧光蛋白基因(gfp)表达载体质粒pDHt-bar-gfp转入DZJ07菌株中,获得了带绿色荧光且遗传稳定的转化子,比较了荧光转化子DZJ07-6和原始菌株在菌落形态、生长特性、拮抗特性以及对小麦纹枯病防治效果等方面的特性,结果发现,荧光转化子DZJ07-6和原始菌株在上述特性方面无明显差别。在此基础上,温室条件下研究了荧光转化子DZJ07-6在小麦植株的定殖动态,荧光显微观察发现荧光转化子DZJ07-6可侵染小麦根系,在小麦根和茎组织的细胞间隙和细胞内大量定殖,平板稀释结果表明,荧光转化子DZJ07-6在小麦根际、根和茎内均能定殖,截止生长的第34天,小麦根际、根内以及茎组织的DZJ07-6菌量分别为2.72×10~6,0.62×10~4,2.26×10~3cfu/g。结果为深入研究DZJ07菌株内生性及生防作用机理奠定了基础。

Colletotrichum sp.DZJ07,a biological control strain isolated from the Eucommia ulmoides,shows potential for control of wheat sharp eyespot.To elucidate the ecological adaptability of DZJ07 of wheat,the green fluorescent protein gene（gfp） was inserted into the DZJ07 genome by Agrobacterium-mediated transformation method,and a GFP-tagged DZJ07-6 strain was constructed.Colony shape,growth rate,antagonistic effect and efficacy for biological control of wheat sharp eyespot were analyzed with DZJ07 and the GFP-tagged strain DZJ07-6.No significant differences were obtained for any of the characteristics tested between the two strains.Then colonization ability of the GFPtagged strain DZJ07-6 on wheat was assessed under greenhouse conditions.And observing by fluorescence microscope showed that the marked strain was able to colonize in the root and stem of wheat,and the population of DZJ07-6 in wheat rhizosphere,root and stem after 34 days were 2.72 × 10^6,0.62 × 10^4 and 2.26 × 10^3 cfu/g respectively.The results afforded scientific basis for its colonization and the further research on bio-controlling mechanism.