The plasmid of pCDMARUBA-Hyg, which contained two insect-resistance genes, sbk （modified from CrylA（c）） and sck （modified from CpTI）, was transformed into an Agrobacterium EHA105 for infection of the calli of a ...The plasmid of pCDMARUBA-Hyg, which contained two insect-resistance genes, sbk （modified from CrylA（c）） and sck （modified from CpTI）, was transformed into an Agrobacterium EHA105 for infection of the calli of a super japonica rice Nanjing 45. Primarily, using polymerase chain reaction （PCR） detection with the primers of sbk and sck genes, 42 positive transgenic plants that were marker-free and contained the two target genes were selected from 97 regenerated plants. Results of southern-blotting indicated that 23, 11, 5, 2 and 1 plants had one, two, three, four and five copies of the transformed genes, respectively. Analysis of reverse transcription PCR （RT-PCR） and Bt gene testing paper showed that 28 T3 generation plants derived from four transgenic plants having a single copy were insect-resistant. Feeding experiment with rice stem borer revealed that the insect resistance was greatly increased with the larva mortality ranging from 94% to 100%. In addition, among the transgenic plants, three T3 transgenic plants possessed some desirable characteristics for breeding and production, such as plant height, seed-setting rate, 1000-grain weight and larva mortality. The mechanism of insect resistance of Bt .qene and its application in rice transgenic research were also briefly discussed.展开更多
The feeding and oviposition behavior of the brown planthopper (BPH), Nilaparvata lugens on two transgenic indica rice homogenous genotypes (B1 and B6) with cry1Ab gene from Bacillus thuringiensis and trans...The feeding and oviposition behavior of the brown planthopper (BPH), Nilaparvata lugens on two transgenic indica rice homogenous genotypes (B1 and B6) with cry1Ab gene from Bacillus thuringiensis and transgenic restored line of hybrid rice (MSA) with SCK gene (a modified CpTI gene) were measured, compared with those on their corresponding non transgenic parental cultivars Jiazao935 and Minghui86 performed by BPH. Under the selection condition of host plants by BPH, loading percentage, oviposition preference and laying egg number of BPH both on transgenic cry1Ab rice and transgenic SCK rice were not significantly different from those on their controls, while their total number of probing wound caused by PBH expect for feeding on B1 plants was markedly more than that on the control. In contrast, under the non selection condition, total number of probing wound caused by BPH on either transgenic cry1Ab rice or transgenic SCK rice was pronouncedly more than those on their controls. Conversely, their honeydew amount excreted by BPH after feeding for 24 h was significantly less than those on the control. As a conclusion, three tested transgenic rice genotypes with insect resistance acted adverse effect on BHP feeding, and no marked effect on BPH oviposition.展开更多
基金supported by the Natural Science Foundation of Jiangsu Province, China (Grant No. BK2008348)the China National Science & Technology Major Project for Breeding New Plant Varieties of Genetically Modified Organisms (GMOs) (Grant No. 2008ZX08001-004)
文摘The plasmid of pCDMARUBA-Hyg, which contained two insect-resistance genes, sbk （modified from CrylA（c）） and sck （modified from CpTI）, was transformed into an Agrobacterium EHA105 for infection of the calli of a super japonica rice Nanjing 45. Primarily, using polymerase chain reaction （PCR） detection with the primers of sbk and sck genes, 42 positive transgenic plants that were marker-free and contained the two target genes were selected from 97 regenerated plants. Results of southern-blotting indicated that 23, 11, 5, 2 and 1 plants had one, two, three, four and five copies of the transformed genes, respectively. Analysis of reverse transcription PCR （RT-PCR） and Bt gene testing paper showed that 28 T3 generation plants derived from four transgenic plants having a single copy were insect-resistant. Feeding experiment with rice stem borer revealed that the insect resistance was greatly increased with the larva mortality ranging from 94% to 100%. In addition, among the transgenic plants, three T3 transgenic plants possessed some desirable characteristics for breeding and production, such as plant height, seed-setting rate, 1000-grain weight and larva mortality. The mechanism of insect resistance of Bt .qene and its application in rice transgenic research were also briefly discussed.
文摘The feeding and oviposition behavior of the brown planthopper (BPH), Nilaparvata lugens on two transgenic indica rice homogenous genotypes (B1 and B6) with cry1Ab gene from Bacillus thuringiensis and transgenic restored line of hybrid rice (MSA) with SCK gene (a modified CpTI gene) were measured, compared with those on their corresponding non transgenic parental cultivars Jiazao935 and Minghui86 performed by BPH. Under the selection condition of host plants by BPH, loading percentage, oviposition preference and laying egg number of BPH both on transgenic cry1Ab rice and transgenic SCK rice were not significantly different from those on their controls, while their total number of probing wound caused by PBH expect for feeding on B1 plants was markedly more than that on the control. In contrast, under the non selection condition, total number of probing wound caused by BPH on either transgenic cry1Ab rice or transgenic SCK rice was pronouncedly more than those on their controls. Conversely, their honeydew amount excreted by BPH after feeding for 24 h was significantly less than those on the control. As a conclusion, three tested transgenic rice genotypes with insect resistance acted adverse effect on BHP feeding, and no marked effect on BPH oviposition.
