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genetic transformation of Tobacco with the Trehalose Synthase Gene from Grifola frondosa Fr. Enhances the Resistance to Drought and Salt in Tobacco

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Journal of Integrative Plant Biology 2005年第5期47卷 579-587页

作者： Shu-ZhenZHANG Ben-PengYANG Cui-LianFENG Huo-LongTANG InstituteofTropicalBiotechnology ChineseAcademyofTropicalAgriculturalSciencesHaikou571101China StateKeyLaboratoryofTropicalCropBiotechnology ChineseAcademyofTropicalAgriculturalSciencesHaikou571101China／／InstituteofTropicalBiotechnologyChineseAcademyofTropicalAgriculturalSciencesHaikou571101China

Trehalose is a non-reducing disaccharide of glucose that functions as a protectant in the stabilization of biological structures and enhances the tolerance of organisms to abiotic stress. In the present study, we report on the expression of the Grifola frondosa Fr. Trehalose synthase (Tsase) gene for manipulating abiotic stress tolerance in tobacco (Nicotiana tabaccum L.). The expression of the transgene was under the control of two tandem copies of the CaMV3 5 S promoter and was transferred into tobacco by Agrobacterium tumefaciens EHA105. Compared with non-transgenic plants, transgenic plants were able to accumulate high levels of products of trehalose, which were increased up to 2.126–2.556 mg/g FW, although levels were undetectable in non-transgenic plants. This level of trehalose in transgenic plants was 400-fold higher than that of transgenic tobacco plants cotransformed with Escherichia coli TPS and TPP on independent expression cassettes, twofold higher than that of transgenic rice plants transformed with a bi functional fusion gene (TPSP) of the trehalose-6-phosphate (T-6-P) synthase (TPS) and T-6-P phosphatase (TPP) of E. coli, and 12-fold higher than that of transgenic tobacco plants transformed the yeast TPS1 gene. It has been reported that transgenic plants with E. coli TPS and/or TPP were severely stunted and had morphological alterations of their roots. Interestingly, our transgenic plants have obvious morphological changes, including thick and deep-coloured leaves, but show no growth inhibition; moreover, these morphological changes can restore to normal type in T2 progenies. Trehalose accumulation in 35S–35S:Tsase plants resulted in increased tolerance to drought and salt, as shown by the results of tests on drought, salt tolerance, and drought physiological indices, such as water content in excised leaves, malondialdehyde content, chlorophyll a and b contents, and the activity of superoxide dismutase and peroxidase in excised leaves. These results

关键词： drought and salt tolerance genetic transformation Grifola frondosa Fr. Nicotiana tabaccum L. trehalose synthase gene

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genetic transformation of marine cyanobacterium Synechococcus ***9311(Cyanophyceae) by electroporation

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Chinese Journal of Oceanology and Limnology 2013年第2期31卷 416-420页

作者：陈华新林瀚智姜鹏李富超秦松 Key Laboratory of Experimental Marine Biology Institute of OceanologyChinese Academy of Sciences Graduate University of Chinese Academy of Sciences Yantai Institute of Coastal Zone Research Chinese Academy of Sciences

Synechococcus ***9311 is a marine cyanobacterium characterized by type IV chromatic acclimation(CA).A genetic transformation system was developed as a first step to elucidate the molecular mechanism of *** results show that Synechococcus ***9311 cells were sensitive to four commonly used antibiotics:ampicillin,kanamycin,spectinomycin,and *** integrative plasmid to disrupt the putative phycoerythrin lyase gene mpeV,using a kanamycin resistance gene as selectable marker,was constructed by recombinant polymerase chain *** plasmid was then transformed into Synechococcus ***9311 via *** transformation efficiency was achieved at a field strength of 2 kV/*** analysis showed that mpeV was fully disrupted following challenge of the transformants with a high concentration of *** addition,the transformants that displayed poor growth on agar SN medium could be successfully plated on agarose SN medium.

关键词： chromatic acclimation electroporation genetic transformation Synechococcus sp. CC9311

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genetic transformation of Nannochloropsis oculata with a Bacterial Phleomycin Resistance Gene as Dominant Selective Marker

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Journal of Ocean University of China 2016年第2期15卷 351-356页

作者： MA Xiaolei PAN Kehou ZHANG Lin ZHU Baohua YANG Guanpin ZHANG Xiangyang Foundation College Jining Medical University Laboratory of Applied Microalgae Biology Key Laboratory of Aquaculture of Ministry of EducationOcean University of China

The gene ble from Streptoalloteichus hindustanus is widely used as a selective antibiotic marker. It can control the phleomycin resistance, and significantly increase the tolerance of hosts to zeocin. The unicellular marine microalga Nannochloropsis oculata is extremely sensitive to zeocin. We selected ble as the selective marker for the genetic transformation of N. oculata. After the algal cells at a density of 2× 10^7 cells mL-1 was digested with 4% hemicellulase and 2% driselase for 1 h, the protoplasts accounted for 90% of the total. The ble was placed at the downstream of promoter HSP70A-RUBS2 isolated from Chlamydomonas reinhardtii, yielding a recombinant expression construct pMS 188. The construct was transferred into the protoplasts through electroporation （1 kV, 15 μS）. The transformed protoplasts were cultured in fresh f/2 liquid medium, and selected on solid f/2 medium supplemented with 500 ng mL-t zeoein. The PCR result proved that ble existed in the transformants. Three transformants had been cultured for at least 5 generations without losing ble. Southern blotting analysis showed that the ble has been integrated into the genome of N. oculata. The ble will serve as a new dominant selective marker in genetic engineering N. oculata.

关键词： Nannochloropsis oculata genetic transformation ble electroporation

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genetic transformation of loblolly pine using mature zygotic embryo explants by Agrobacterium tumefaciens

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Journal of Forestry Research 2000年第4期11卷 215-222页

作者：唐巍 North Carolina State University Forest Biotechnology Group Department of Forestry Raleigh USA (GRID:grid.40803.3f) (ISNI:***)

Agrobacterium tumefaciens strain LBA 4404 carrying pBI121 plasmid was used to transform mature zygotic embryos of three genotypes (E-Hb, E-Ma, and E-Mc) of loblolly pine. The results demonstrated that the expression frequency of (-glucuronidase reporter gene (GUS) varied among genotypes after mature zygotic embryos were infected with Agrobacterium tumefaciens cultures. The highest frequency (27.8%) of GUS expressing embryos was obtained from genotype E-Mc with mean number of 21.9 blue GUS spots per embryo. Expression of (-glucuronidase reporter gene was observed on cotyledons, hypocotyls, and radicles of transformed mature zygotic embryos, as well as on organogenic callus and regenerated shoots derived from co-cultivated mature zygotic embryos. Nineteen regenerated transgenic plants were obtained from GUS expression and kanamycin resistant calli. The presence and integration of the GUS gene was confirmed by polymerase chain reaction (PCR) and Southern blot analysis. These results suggested that an efficient Agrobacterium tumefaciens-mediated transformation protocol for stable integration of foreign genes into loblolly pine has been developed and that this transformation system could be useful for the future studies on transferring economically important genes to loblolly pine.

关键词： Pinus taeda L. genetic transformation Agrobacterium tumefaciens (-glucuronidase gene Polymerase chain reaction Southern blot

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Cassava genetic transformation and its Application in Breeding

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Journal of Integrative Plant Biology 2011年第7期53卷 552-569页

作者： Jia Liu Qijie Zheng Qiuxiang Ma Kranthi Kumar Gadidasu Peng Zhang National Key Laboratory of PlantMolecular genetics institute of Plant Physiology and Ecology Shanghai Institutes for Biological Sciences Chinese Academy of Sciences Shanghai 200032 China Key Laboratory of Synthetic Biology Institute of Plant Physiology and Ecology Shanghai Institutes for Biological Sciences Chinese Academy of Sciences Shanghai 200032 China plant Biotechnology Research Unit Department of Biotechnology Kakatiya University Warangal 506009 India Shanghai Chenshan Plant Science Research Center Chinese Academy of Sciences Shanghai Chenshan Botanical Garden Shangha 201602. China

As a major source of food, cassava （Manihot esculenta Crantz） is an important root crop in the tropics and subtropics of Africa and Latin America, and serves as raw material for the production of starches and bioethanol in tropical Asia. Cassava improvement through genetic engineering not only overcomes the high heterozygosity and serious trait separation that occurs in its traditional breeding, but also quickly achieves improved target traits. Since the first report on genetic transfor- mation in cassava in 1996, the technology has gradually matured over almost 15 years of development and has overcome cassava genotype constraints, changing from mode cultivars to farmer-preferred ones. Significant progress has been made in terms of an increased resistance to pests and diseases, biofortification, and improved starch quality, building on the fundamental knowledge and technologies related to planting, nutrition, and the processing of this important food crop that has often been neglected. *** has great potential-in food security and bioenergy development worldwide.

关键词： Application genetic transformation Manihot esculenta Crantz molecular breeding.

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Current status of genetic transformation technology developed in cucumber(Cucumis sativus L.)

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Journal of Integrative Agriculture 2015年第3期14卷 469-482页

作者： WANG Shun-li Seong Sub Ku YE Xing-guo HE Cong-fen Suk Yoon Kwon Pil Son Choi Key Laboratory of Biology and genetic Improvement of Horticultural Crops Ministry of Agriculture/Institute of Vegetables and Flowers Chinese Academy of Agricultural Sciences Department of Oriental Pharmaceutical Development Nambu University Institute of Crop Science Chinese Academy of Agricultural Sciences/National Key Facility for Crop Gene Resources and Genetic Improvement School of Sciences Beijing Technology and Business University Plant Systems Engineering Research Center

genetic transformation is an important technique for functional genomics study and genetic improvement of plants. Until now, Agrobacterium-mediated transformation methods using cotyledon as explants has been the major approach for cucumber, and its frequency has been up to 23%. For example, significantly enhancement of the transformation efficiency of this plant species was achieved from the cotyledon explants of the cultivar Poinsett 76 infected by Agrobacterium strains EHA105 with efficient positive selection system in lots of experiments. This review is to summarize some key factors influencing cucumber regeneration and genetic transformation, including target genes, selection systems and the ways of transgene introduction, and then to put forward some strategies for the increasing of cucumber transformation efficiency. In the future, it is high possible for cucumber to be potential bioreactor to produce vaccine and biomaterials for human beings.

关键词： Cucumis sativus L. plant regeneration genetic transformation positive selection system

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Simultaneous genetic transformation and genome editing of mixed lines in soybean(Glycine max)and maize(Zea mays)

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aBIOTECH 2024年第2期5卷 169-183页

作者： Michelle Valentine David Butruille Frederic Achard Steven Beach Brent Brower-Toland Edward Cargill Megan Hassebrock Jennifer Rinehart Thomas Ream Yurong Chen Bayer Crop Science 700 Chesterfield Parkway WChesterfieldMO 63017USA

Robust genome editing technologies are becoming part of the crop breeding ***,genome editing is usually conducted either at a single locus,or multiple loci,in a variety at one *** parallel genomics platforms,multifaceted genome editing capabilities,and flexible transformation systems enable targeted variation at nearly any locus,across the spectrum of genotypes within a *** demonstrate here the simultaneous transformation and editing of many genotypes,by targeting mixed seed embryo explants with genome editing machinery,followed by re-identification through genotyping after plant *** and Editing of Mixed Lines(TREDMIL)produced transformed individuals representing 101 of 104(97%)mixed elite genotypes in soybean;and 22 of 40(55%)and 9 of 36(25%)mixed maize female and male elite inbred genotypes,*** of edited genotypes for the regenerated individuals identified over 800 distinct edits at the Determinate1(Dt1)locus in samples from 101 soybean genotypes and 95 distinct Brown midrib3(Bm3)edits in samples from 17 maize *** results illustrate how TREDMIL can help accelerate the development and deployment of customized crop varieties for future precision breeding.

关键词： Maize Soybean Seed embryo explants TREDMIL genetic transformation Genome editing

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Enhancing wheat regeneration and genetic transformation through overexpression of TaLAX1

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Plant Communications 2024年第5期5卷 9-24页

作者： Yang Yu Haixia Yu Jing Peng Wang Jinsong Yao Yi Peng Wang Feng Li Zhang Shi Rong Wang Yajie Zhao Xiang Yu Zhao Xian Sheng Zhang Ying Hua Su State Key Laboratory of Crop Biology College of Life SciencesShandong Agricultural UniversityTai’anShandong 271018China

In the realm of genetically transformed crops,the process of plant regeneration holds utmost ***,the low regeneration efficiency of several wheat varieties currently restricts the use of genetic transformation for gene functional analysis and improved crop *** research explores overex-pression of TaLAX PANICLE1(TaLAX1),which markedly enhances regeneration efficiency,thereby boost-ing genetic transformation and genome editing in *** noteworthy is the substantial increase in regeneration efficiency of common wheat varieties previously regarded as recalcitrant to genetic *** study shows that increased expression of TaGROWTH-REGULATING FACTOR(TaGRF)genes,alongside that of their co-factor,TaGRF-INTERACTING FACTOR 1(TaGIF1),enhances cytokinin accumulation and auxin response,which may play pivotal roles in the improved regeneration and transfor-mation of TaLAX1-overexpressing wheat *** of TaLAX1 homologs also significantly in-creases the regeneration efficiency of maize and soybean,suggesting that both monocot and dicot crops can benefit from this *** shed light on a gene that enhances wheat genetic trans-formation and elucidate molecular mechanisms that potentially underlie wheat regeneration.

关键词： plant regeneration genetic transformation TaLAX1 TaGRF4–TaGIF1 cytokinin auxin wheat

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Advancements in plant regeneration and genetic transformation of grapevine(Vitis spp.)

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Journal of Integrative Agriculture 2021年第6期20卷 1407-1434页

作者： ZHANG Xiu-ming WU Yi-fei LI Zhi SONG Chang-bing WANG Xi-ping Key Laboratory of Horticultural Plant Biology and Germplasm Innovation in Northwest China Ministry of Agriculture/State Key Laboratory of Crop Stress Biology in Arid Areas/College of HorticultureNorthwest A&F UniversityYang ling 712100P.R.China College of Biological Science and Engineering North Minzu UniversityYinchuan 750021P.R.China

Grapevine(Vitis spp.)is one of the most economically important fruit crops worldwide,and there is considerable interest in improving its major agronomic and enological traits in response to ever-changing agricultural environments and consumer *** genetic techniques in particular,associated with rapid technological advancements,provide an attractive alternative to conventional breeding approaches for developing new grapevine varieties with enhanced yield performance,quality,stress tolerance and disease *** date,several grapevine varieties have been transformed with genes associated with diverse functions through biolistic bombardment and/or Agrobacterium-mediated transformation,and transgenic grape lines have been obtained using established regeneration ***,a wide range of factors,including genotype,explant source and culture medium,have been shown to affect the efficiency of plant ***,the selection and use of acceptor materials,bacterial strain and cell density,selectable markers and selection methods also influence transformation *** paper provides an overview of recent advances in grapevine regeneration and genetic transformation and in-depth discussion of the major limiting factors,and discusses promising future strategies to develop robust plant regeneration and genetic transformation in grapevine.

关键词： grapevine organogenesis somatic embryogenesis plant regeneration genetic transformation

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Establishment of A Simple and Efficient Agrobacterium-mediated genetic transformation System to Chinese Cabbage(Brassica rapa ***)

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Horticultural Plant Journal 2021年第2期7卷 117-128页

作者： Xiaonan Li Haiyan Li Yuzhu Zhao Peixuan Zong Zongxiang Zhan Zhongyun Piao Department of Horticulture Shenyang Agricultural UniversityShenyangLiaoning 110866China

Chinese cabbage,belonging to Brassica rapa species,is an important vegetable in Eastern *** is well known that Chinese cabbage is quite recalcitrant to genetic transformation and the transgenic frequency is generally *** lack of an efficient and stable genetic transformation system for Chinese cabbage has largely limited related gene functional *** this study,we firstly developed a regeneration system for Chinese cabbage by optimizing numerous factors,with 93.50%regeneration rate on *** on this,a simple and efficient Agrobacteriummediated genetic transformation methodwas established,without pre-culture procedure and concentration adjustment of hormone and AgNO_(3) in co-cultivation and selection *** this system,transformants could be obtained within 3.5–4.0 *** transformation frequency is up to 10.83%.The establishment of this simple and efficient genetic transformation method paved the way for further gene editing and functional studies in Chinese cabbage.

关键词： Chinese cabbage Agrobacterium tumefaciens genetic transformation

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