Expression vector pE14 with double resistance to virus and insect was constructed by inserting CW-cp gene and Bt-toxin gene one by into T-DNA of the same binary vector pE3. Tobacco was then transformed with Agrbacter...Expression vector pE14 with double resistance to virus and insect was constructed by inserting CW-cp gene and Bt-toxin gene one by into T-DNA of the same binary vector pE3. Tobacco was then transformed with Agrbacterium tumefaciens (At)GV311-SE carrying PE14. Nopaline assay and PCR amplification confirmed that both CW-cp gene and Bt-toxin gene had been introduced into tobacco genome by T-DNA of PE3. Test attack with virus and demonstrated, in some of the transgenic plants, double resistance to both infection by CMV and damage by Manduca sexta.展开更多
Chromium(Cr) may cause losses in the yield of field plant, which is one of the favorite habitats of Bacillus thuringiensis(Bt). The purposes of our study were to assess the Cr(VI)-resistance and Cr(VI)-reducing abilit...Chromium(Cr) may cause losses in the yield of field plant, which is one of the favorite habitats of Bacillus thuringiensis(Bt). The purposes of our study were to assess the Cr(VI)-resistance and Cr(VI)-reducing abilities of an indigenous soil isolate of Bt and to determine the factors governing Cr(VI) reduction. Towards this end a novel dichromate-reducing Bt BRC-ZYR2, characterized with insecticidal crystal proteins(ICPs), was isolated from a uranium deposit. Minimum inhibitory concentrations(MICs) of Cr(VI) were determined by broth dilution method and the concentrations of Cr(VI) and total Cr in the supernatant were quantified colorimetrically using 1,5-diphenylcarbazide(DPC) reagent and a mixture of sulfuric-nitric acids, respectively. The isolate contained five ICP genes(cry1Ba, cry1 Bb, cry1Be/cry1 Bf, cry9 Ca and cry9Da) and exhibited a high level of Cr(VI) resistance with MICs of 150 mg L-1at pH 7.0 and 30?C, and 500 mg L-1under optimal conditions(pH 9.0 and 40?C). The total Cr concentration was similar to initial concentration of Cr(VI) under the optimal condition, suggesting that the essential removal of the Cr(VI) was dependent on Bt reduction. Under optimal conditions, the initial Cr(VI) concentrations from 25 to 75 mg L-1significantly decreased in 24 h after incubation. Addition of Mn2+, Co2+, Mo2+and Cu2+activated Bt-mediated Cr(VI) reduction, while Zn2+, Ni2+and glucose were found to inhibit the reduction. Our results indicated that this isolate could be a promising biopesticide with the potential for both insect biocontrol and Cr bioremediation in the field.展开更多
文摘Expression vector pE14 with double resistance to virus and insect was constructed by inserting CW-cp gene and Bt-toxin gene one by into T-DNA of the same binary vector pE3. Tobacco was then transformed with Agrbacterium tumefaciens (At)GV311-SE carrying PE14. Nopaline assay and PCR amplification confirmed that both CW-cp gene and Bt-toxin gene had been introduced into tobacco genome by T-DNA of PE3. Test attack with virus and demonstrated, in some of the transgenic plants, double resistance to both infection by CMV and damage by Manduca sexta.
基金Supported by the National Science and Technology Project in Rural Area during the"2th Five-Year Plan",China(Nos.2011AA10A203 and 2011BAD03B02-2)the National Natural Science Foundation of China(No.31201574)+3 种基金the Project of Fujian Provincial Department of Science and Technology,China(No.2011N5003)the State Key Laboratory for Biology of Plant Diseases and Insect Pests,Institute of Plant Protection,Chinese Academy of Agricultural Sciences(No.SKL2012OP05)the Talent Training Program for Distinguished Young Scholars in Universities of Fujian Province,China(No.JA12092)the Science Fund for Distinguished Young Scholars of Fujian Agriculture and Forestry University,China(No.xjq201203)
文摘Chromium(Cr) may cause losses in the yield of field plant, which is one of the favorite habitats of Bacillus thuringiensis(Bt). The purposes of our study were to assess the Cr(VI)-resistance and Cr(VI)-reducing abilities of an indigenous soil isolate of Bt and to determine the factors governing Cr(VI) reduction. Towards this end a novel dichromate-reducing Bt BRC-ZYR2, characterized with insecticidal crystal proteins(ICPs), was isolated from a uranium deposit. Minimum inhibitory concentrations(MICs) of Cr(VI) were determined by broth dilution method and the concentrations of Cr(VI) and total Cr in the supernatant were quantified colorimetrically using 1,5-diphenylcarbazide(DPC) reagent and a mixture of sulfuric-nitric acids, respectively. The isolate contained five ICP genes(cry1Ba, cry1 Bb, cry1Be/cry1 Bf, cry9 Ca and cry9Da) and exhibited a high level of Cr(VI) resistance with MICs of 150 mg L-1at pH 7.0 and 30?C, and 500 mg L-1under optimal conditions(pH 9.0 and 40?C). The total Cr concentration was similar to initial concentration of Cr(VI) under the optimal condition, suggesting that the essential removal of the Cr(VI) was dependent on Bt reduction. Under optimal conditions, the initial Cr(VI) concentrations from 25 to 75 mg L-1significantly decreased in 24 h after incubation. Addition of Mn2+, Co2+, Mo2+and Cu2+activated Bt-mediated Cr(VI) reduction, while Zn2+, Ni2+and glucose were found to inhibit the reduction. Our results indicated that this isolate could be a promising biopesticide with the potential for both insect biocontrol and Cr bioremediation in the field.
