Chloroplast genetic engineering, with several advantages over nuclear genetic engineering, is now regarded as an attractive new technology in basic and applied research, including deepening our understanding of plasti...Chloroplast genetic engineering, with several advantages over nuclear genetic engineering, is now regarded as an attractive new technology in basic and applied research, including deepening our understanding of plastid genome, engineering plant metabolic system, generating transplastomic plants with higher resistance to insect, disease, drought and herbicide and bioproducing of antibodies and vaccines. In this review, the principle and operating system for chloroplast genetic engineering and its application in higher plants have been discussed.展开更多
Plastid transformation offers valuable benefits in plant biotechnology,such as high-level transgene expression and the absence of gene silencing.Here we describe the first protocol of a plastid transformation system f...Plastid transformation offers valuable benefits in plant biotechnology,such as high-level transgene expression and the absence of gene silencing.Here we describe the first protocol of a plastid transformation system for a woody vine(liana)kiwifruit(Actinidia chinensis).The transgenic DNA carries a spectinomycin-resistance gene(aadA)cassette and a green fluorescent protein(GFP)reporter gene cassette,flanked by two adjacent kiwifruit plastid genome sequences,thereby allowing targeted insertion between the trnfM and trnG genes.Six spectinomycin-resistant shoots were obtained out of 12 plates subjected to bombardment,and two were positive events,confirmed through PCR and Southern blot analyses.The GFP was localized to plastids as monitored by confocal laser scanning microscopy and reached 2.5%of leaf total soluble protein.Success in kiwifruit extends transplastomic technology of woody species beyond poplar,and will provide an attractive biosynthetic chassis for molecular farming.展开更多
Plant derived protease inhibitors(PIs)are a promising defensin for crop im-provement and insect pest management.Although agronomist made significant efforts in utilizing PIs for managing insect pests.the potentials of...Plant derived protease inhibitors(PIs)are a promising defensin for crop im-provement and insect pest management.Although agronomist made significant efforts in utilizing PIs for managing insect pests.the potentials of PIs are still obscured.Insect ability to compensate nutrient starvation induced by dietary PI feeding using different strategies,that is,overexpression of PI-sensitive protease,expression of PI-insensitive proteases,degradation of PI,has made this innumerable collection of PIs worthless.A practical challenge for agronomist is to identify potent PI candidates,to limit insect compensatory responses and to elucidate insect compensatory and resistance mechanisms activated upon herbivory.This knowledge could be further efficiently utilized to identify potential targets for RNAi-mediated pest control.These vital genes of insects could be functionally anno-tated using the advanced gene-editing technique,CRISPR/Cas9.Contemporary research is exploiting different in silico and modern molecular biology techniques to utilize PIs in controlling insect pests efficiently.This review is structured to update recent advancements in this field,along with is chronological background.展开更多
Chloroplast integration and expression vector containing expression cassettes for phbB, phbA, phbC and aadA genes was constructed and bombarded into the tobacco chloroplast genome. Transplastomic plants were analyzed ...Chloroplast integration and expression vector containing expression cassettes for phbB, phbA, phbC and aadA genes was constructed and bombarded into the tobacco chloroplast genome. Transplastomic plants were analyzed with PCR and Southern blot. Their homoplastomy was also judged. Northern dot and RT-PCR analysis were employed to investigate transgene expression at transcriptional level. The results indicate that the chloroplast transformation system is compatible for poly-3-hydroxybutyrate (PHB) production.展开更多
文摘Chloroplast genetic engineering, with several advantages over nuclear genetic engineering, is now regarded as an attractive new technology in basic and applied research, including deepening our understanding of plastid genome, engineering plant metabolic system, generating transplastomic plants with higher resistance to insect, disease, drought and herbicide and bioproducing of antibodies and vaccines. In this review, the principle and operating system for chloroplast genetic engineering and its application in higher plants have been discussed.
基金supported by grants from the National Natural Science Foundation of China(32271912,32071477).
文摘Plastid transformation offers valuable benefits in plant biotechnology,such as high-level transgene expression and the absence of gene silencing.Here we describe the first protocol of a plastid transformation system for a woody vine(liana)kiwifruit(Actinidia chinensis).The transgenic DNA carries a spectinomycin-resistance gene(aadA)cassette and a green fluorescent protein(GFP)reporter gene cassette,flanked by two adjacent kiwifruit plastid genome sequences,thereby allowing targeted insertion between the trnfM and trnG genes.Six spectinomycin-resistant shoots were obtained out of 12 plates subjected to bombardment,and two were positive events,confirmed through PCR and Southern blot analyses.The GFP was localized to plastids as monitored by confocal laser scanning microscopy and reached 2.5%of leaf total soluble protein.Success in kiwifruit extends transplastomic technology of woody species beyond poplar,and will provide an attractive biosynthetic chassis for molecular farming.
文摘Plant derived protease inhibitors(PIs)are a promising defensin for crop im-provement and insect pest management.Although agronomist made significant efforts in utilizing PIs for managing insect pests.the potentials of PIs are still obscured.Insect ability to compensate nutrient starvation induced by dietary PI feeding using different strategies,that is,overexpression of PI-sensitive protease,expression of PI-insensitive proteases,degradation of PI,has made this innumerable collection of PIs worthless.A practical challenge for agronomist is to identify potent PI candidates,to limit insect compensatory responses and to elucidate insect compensatory and resistance mechanisms activated upon herbivory.This knowledge could be further efficiently utilized to identify potential targets for RNAi-mediated pest control.These vital genes of insects could be functionally anno-tated using the advanced gene-editing technique,CRISPR/Cas9.Contemporary research is exploiting different in silico and modern molecular biology techniques to utilize PIs in controlling insect pests efficiently.This review is structured to update recent advancements in this field,along with is chronological background.
基金This work was supported by the State "863" High-Tech Program (Grant No. 101-04-03-04)the National Natural Science Foundation of China (Grant No. 59673008).
文摘Chloroplast integration and expression vector containing expression cassettes for phbB, phbA, phbC and aadA genes was constructed and bombarded into the tobacco chloroplast genome. Transplastomic plants were analyzed with PCR and Southern blot. Their homoplastomy was also judged. Northern dot and RT-PCR analysis were employed to investigate transgene expression at transcriptional level. The results indicate that the chloroplast transformation system is compatible for poly-3-hydroxybutyrate (PHB) production.
