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Advances in plastid transformation for metabolic engineering in higher plants

aBIOTECH 2022年第3期3卷 224-232页

作者： Sheng Yang Yi Deng Shengchun Li State Key Laboratory of Biocatalysis and Enzyme Engineering School of Life SciencesHubei UniversityWuhan430062China

The plastid(chloroplast)genome of higher plants is an appealing target for metabolic engineering via genetic transformation.Although the bacterial-type plastid genome is small compared with the nuclear genome,it can accommodate large quantities of foreign genes that precisely integrate through homologous recombination.Engineering complex metabolic pathways in plants often requires simultaneous and concerted expression of multiple transgenes,the possibility of stacking several transgenes in synthetic operons makes the transplastomic approach amazing.The potential for extraordinarily high-level transgene expression,absence of epigenetic gene silencing and transgene containment due to the exclusion of plastids from pollen transmission in most angiosperm species further add to the attractiveness of plastid transformation technology.This minireview describes recent advances in expanding the toolboxes for plastid genome engineering,and highlights selected high-value metabolites produced using transplastomic plants,including artemisinin,astaxanthin and paclitaxel.

关键词： plastid transformation Metabolic engineering Multiple transgenes High-level expression

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Complete protection from Henosepilachna vigintioctopunctata by expressing long double-stranded RNAs in potato plastids

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Journal of Integrative Plant Biology 2023年第4期65卷 1003-1011页

作者： Wenbo Xu Miao Zhang Yangcun Li Wanwan He Shengchun Li Jiang Zhang State Key Laboratory of Biocatalysis and Enzyme Engineering Hubei Hongshan LaboratorySchool of Life SciencesHubei UniversityWuhan 430062China Agricultural Genomics Institute at Shenzhen Chinese Academy of Agricultural SciencesShenzhen 518000China

RNA interference(RNAi)has emerged as a powerful technology for pest management.Previously,we have shown that plastid-mediated RNAi(PM-RNAi)can be utilized to control the Colorado potato beetle,an insect pest in the Chrysomelidae family;however,whether this technology is suitable for controlling pests in the Coccinellidae remained unknown.The coccinellid 28-spotted potato ladybird(Henosepilachna vigintioctopunctata;HV)is a serious pest of solanaceous crops.In this study,we identified three efficient target genes(β-Actin,SRP54,and SNAP)for RNAi using in vitro double-stranded RNAs(dsRNAs)fed to HV,and found that dsRNAs targetingβ-Actin messenger RNA(dsACT)induced more potent RNAi than those targeting the other two genes.We next generated transplastomic and nuclear transgenic potato(Solanum tuberosum)plants expressing HV dsACT.Long dsACT stably accumulated to up to 0.7%of the total cellular RNA in the transplastomic plants,at least three orders of magnitude higher than in the nuclear transgenic plants.Notably,the transplastomic plants also exhibited a significantly stronger resistance to HV,killing all larvae within 6 d.Our data demonstrate the potential of PM-RNAi as an efficient pest control measure for HV,extending the application range of this technology to Coccinellidae pests.

关键词： double‐stranded RNA Henosepilachna vigintioctopunctata pest control plastid transformation RNA interference

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Control of a sap-sucking insect pest by plastidmediated RNA interference

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Molecular Plant 2022年第7期15卷 1176-1191页

作者： Yi Dong Mengting Wu Qi Zhang Jinqiu Fu F.Vanessa Loiacono Yong Yang Zican Wang Shengchun Li Ling Chang Ralph Bock Jiang Zhang State Key Laboratory of Biocatalysis and Enzyme Engineering Hubei Hongshan LaboratorySchool of Life SciencesHubei UniversityWuhan 430062China Max-Planck-Institut fur Molekulare Pflanzenphysiologie Am Mühlenberg 1D-14476 Potsdam-GolmGermany

Expression of double-stranded RNAs in plastids offers great potential for the efficient control of chewing insects.However,many insect pests do not consume plant tissue but rather feed on the host plant by sucking sap from the vascular system.Whether or not plastid-mediated RNA interference(RNAi)can be employed to control sap-sucking insects is unknown.Here,we show that five species of sap-sucking hemipteran insects acquire plastid RNA upon feeding on plants.We generated both nuclear transgenic and transplastomic tobacco plants expressing double-stranded RNAs targeting the MpDhc64C gene,a newly identified efficient target gene of RNAi whose silencing causes lethality to the green peach aphid Myzus persicae.In a whole-plant bioassay,transplastomic plants exhibited significant resistance to aphids,as evidenced by reduced insect survival,impaired fecundity,and decreased weight of survivors.The protective effect was comparable with that conferred by the best-performing nuclear transgenic plants.We found that the proportion of aphids on mature leaves of transplastomic plants was significantly lower compared with that of nuclear transgenic plants.When aphids were allowed to infest.only the mature leaves,transplastomic plants grew significantly faster and were overall better protected from the pest compared with nuclear transgenic plants.When monitored by electrical-penetration-graph analyses and aphid avoidance response experiments,the insects displayed remarkable alterations in feeding behavior,which was different in nuclear transgenic and transplastomic plants,likely reflecting specific avoidance strategies to toxic RNA molecules.Taken together,our study demonstrates that plastid-mediated RNAi provides an efficient strategy for controlling at least some sap-sucking insect pests,even though there is most likely no or only very little chloroplast RNA in the sap.

关键词： plastid transformation RNA interference pest control sap-sucking insect aphid hemiptera

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The Translational Apparatus of plastids and Its Role in Plant Development

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Molecular Plant 2014年第7期7卷 1105-1120页

作者： Nadine Tiller Ralph Bock Max-Planck-Institut for Molekulare Pflanzenphysiologie Am Muhlenberg 1 D-14476 Potsdam-Golm Germany

Chloroplasts （plastids） possess a genome and their own machinery to express it. Translation in plastids occurs on bacterial-type 70S ribosomes utilizing a set of tRNAs that is entirely encoded in the plastid genome. In recent years, the components of the chloroplast translational apparatus have been intensely studied by proteomic approaches and by reverse genetics in the model systems tobacco （plastid-encoded components） and Arabidopsis （nucleus-encoded components）. This work has provided important new insights into the structure, function, and biogenesis of chloroplast ribosomes, and also has shed fresh light on the molecular mechanisms of the translation process in plastids. In addition, mutants affected in plastid translation have yielded strong genetic evidence for chloroplast genes and gene products influencing plant develop- ment at various levels, presumably via retrograde signaling pathway（s）. In this review, we describe recent progress with the functional analysis of components of the chloroplast translational machinery and discuss the currently available evidence that supports a significant impact of plastid translational activity on plant anatomy and morphology.

关键词： plastid translation ribosome ribosomal protein evolution plastid transformation retrograde signaling leaf development palisade cell.

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