1.Brassica juncea WRKY12 mediates bolting and flowering by interacting with the SOC1 and FUL promoters.
Yifang HUANG ; Yue DONG ; Yue YU ; Dakun LIU ; Qinlin DENG ; Yuanda WANG ; Dayong WEI ; Zhimin WANG ; Qinglin TANG
Chinese Journal of Biotechnology 2025;41(7):2818-2828
Flowering and bolting are important agronomic traits in cruciferous crops such as Brassica juncea. Timely flowering can ensure the crop organ yield and quality, as well as seed propagation. The WRKY family plays an important role in regulating plant bolting and flowering, while the function and mechanism of WRKY12 in B. juncea remain unknown. To explore its function and mechanism in bolting and flowering of B. juncea, we cloned and characterized the BjuWRKY12 gene in B. juncea and found that its expression levels were significantly higher in flowers and inflorescences than in leaves. BjuWRKY12 belonged to the Ⅱc subfamily of the WRKY family, and subcellular localization indicated that the protein was located in the nucleus. Ectopic overexpression of BjuWRKY12 in transgenic lines promoted bolting and flowering, leading to significant increases in the expression levels of flowering integrators SOC1 and FUL. Furthermore, yeast one-hybrid and dual luciferase reporter system assays confirmed that BjuWRKY12 directly bound to the promoters of BjuSOC1 and BjuFUL, undergoing protein-DNA interactions. This discovery gives new insights into the regulation network and molecular mechanisms of BjuWRKY12, laying a theoretical foundation for the breeding of high-yield and high-quality varieties of B. juncea.
Mustard Plant/metabolism*
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Flowers/growth & development*
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Plant Proteins/physiology*
;
Promoter Regions, Genetic/genetics*
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Gene Expression Regulation, Plant
;
Plants, Genetically Modified/genetics*
;
Transcription Factors/metabolism*
;
MADS Domain Proteins/metabolism*
2.Application and prospects of synthetic biology in the genetic improvement of rice.
Luyao TANG ; Yiting WEI ; Yuqing XU ; Yuexing WANG ; Yuchun RAO
Chinese Journal of Biotechnology 2025;41(10):3840-3862
Synthetic biology, recognized as one of the most revolutionary interdisciplinary fields in the 21st century, has established innovative strategies for the genetic improvement of rice through the integration of multidisciplinary technologies including genome editing, genetic circuit design, metabolic engineering, and artificial intelligence. This review systematically summarizes recent research advancements and breakthrough achievements in the application of synthetic biology in the genetic improvement of rice, focusing on three critical domains: yield improvement, nutritional quality fortification, and reinforcement of disease resistance and abiotic stress tolerance. It elucidates that synthetic biology enables precise genomic and metabolic pathway engineering through modular, standard, and systematic approaches, effectively overcoming the limitations of conventional breeding methods characterized by prolonged cycles and restricted trait modification capabilities. The implementation of synthetic biology has facilitated synergistic improvement of multi-traits, thereby providing critical technical references for developing elite rice cultivars with superior productivity and nutritional value. These technological breakthroughs hold significant implications for ensuring global food security and promoting green and sustainable development of agriculture.
Oryza/growth & development*
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Synthetic Biology/methods*
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Metabolic Engineering
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Plant Breeding/methods*
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Gene Editing
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Genetic Engineering/methods*
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Plants, Genetically Modified/genetics*
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Disease Resistance/genetics*
3.Advantages and potential ecological risks of genetically modified crops.
Qingjie CHEN ; Yuqing CHENG ; Yu MA ; Ning XU
Chinese Journal of Biotechnology 2025;41(10):3891-3906
Genetically modified (GM) crops, as a pivotal innovation in modern agriculture, exhibit significant advantages such as pest and disease resistance, herbicide tolerance, stress tolerance, and yield enhancement. However, their widespread adoption has been associated with potential ecological risks, including weediness of transgenic plants, gene flow, emergence of novel viral strains in virus-resistant crops, impacts on non-target organisms and soil ecosystems, and evolution of target pest resistance. This review focuses on the dual characteristics of GM crops, systematically examining their agronomic benefits and the underlying mechanisms of ecological risks. This review provides a theoretical foundation for optimizing the development of GM crops and ecological risk management, facilitating sustainable agricultural practices.
Plants, Genetically Modified/growth & development*
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Crops, Agricultural/growth & development*
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Ecosystem
;
Ecology
4.Disruption of OsRhoGDI2 by CRISPR/Cas9 technology leads to semi-dwarf in rice.
Kaijie WANG ; Wenjing AN ; Yafei LIU ; Di LIU ; Lianjie FENG ; Junjie WANG ; Junjun HUANG ; Xiaofei LIU ; Weihong LIANG
Chinese Journal of Biotechnology 2020;36(4):707-715
OsRhoGDI2 was isolated as a putative partner of Rho protein family member OsRacD from rice panicles by yeast two-hybrid, but its function remains unknown. In order to identify the function of OsRhoGDI2, OsRhoGDI2 knockout mutants were created by CRISPR/Cas9 technology. The results showed that two different homozygous mutants were obtained in T0 generation, and eight kinds homozygous mutants were identified in T1 generation. Sequence analysis revealed that the base substitution or base deletion occurred near the editing targets of the gene in knockout rice, and it could be expected that the truncated OsRhoGDI2 proteins lacking the RhoGDI conserved domain would be generated. Phenotype analysis showed that the OsRhoGDI2 knockout rice plants were significantly lower than the control plants. Statistical analysis confirmed that the significant decrease of plant height was due to the shortening of the second and third internodes, suggesting that OsRhoGDI2 gene may be related with rice height control.
CRISPR-Cas Systems
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Genes, Plant
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genetics
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Oryza
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genetics
;
growth & development
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Plants, Genetically Modified
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rho Guanine Nucleotide Dissociation Inhibitor beta
;
genetics
5.Construction and function of a root-specific promoter SRSP.
Wenwen CUI ; Jing CHI ; Yanfang FENG ; Lili GENG ; Rongmei LIU
Chinese Journal of Biotechnology 2020;36(4):700-706
The responsibility of root is absorbing water and nutrients, it is an important plant tissue, but easily to be affected by biotic and abiotic stresses, affecting crop growth and yield. The design of a synthetic root-specific promoter provides candidate promoters for the functional analysis and efficient expression of stress-related genes in crop roots. In this study, a synthetic root-specific module (pro-SRS) was designed using tandem four-copies of root specific cis-acting elements (OSE1ROOTNODULE, OSE2ROOTNODULE, SP8BFIBSP8AIB, and ROOTMOTIFAPOX1), and fused with minimal promoter from the CaMV 35S promoter to synthesize an artificially synthetic SRSP promoter. The SRSP promoter was cloned in pCAMBIA2300.1 by replacing CaMV 35S promoter so as to drive GUS expression. The constructs with SRSP promoter were transformed in tobacco by Agrobacterium-mediated method. SRSP promoter conferred root-specific expression in transgenic tobacco plants through Real-time PCR (RT-PCR) analysis and GUS histochemical staining analysis. It is indicated that the repeated arrangement of cis-acting elements can realize the expected function of the promoter. This study laid a theoretical foundation for the rational design of tissue-specific promoters.
Agrobacterium
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genetics
;
Cloning, Molecular
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Gene Expression Regulation, Plant
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Plant Roots
;
genetics
;
Plants, Genetically Modified
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Promoter Regions, Genetic
;
genetics
;
Stress, Physiological
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Tobacco
;
genetics
;
growth & development
;
Transformation, Genetic
6.Research progress of genetic engineering on medicinal plants.
China Journal of Chinese Materia Medica 2015;40(4):594-601
The application of genetic engineering technology in modern agriculture shows its outstanding role in dealing with food shortage. Traditional medicinal plant cultivation and collection have also faced with challenges, such as lack of resources, deterioration of environment, germplasm of recession and a series of problems. Genetic engineering can be used to improve the disease resistance, insect resistance, herbicides resistant ability of medicinal plant, also can improve the medicinal plant yield and increase the content of active substances in medicinal plants. Thus, the potent biotechnology can play an important role in protection and large area planting of medicinal plants. In the development of medicinal plant genetic engineering, the safety of transgenic medicinal plants should also be paid attention to. A set of scientific safety evaluation and judgment standard which is suitable for transgenic medicinal plants should be established based on the recognition of the particularity of medicinal plants.
Genetic Engineering
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Plant Diseases
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genetics
;
prevention & control
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Plants, Genetically Modified
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chemistry
;
genetics
;
growth & development
;
metabolism
;
Plants, Medicinal
;
chemistry
;
genetics
;
growth & development
;
metabolism
7.Induction and in vitro culture of hairy roots of Dianthus caryophyllus and its plant regeneration.
Heping SHI ; Yuanfeng ZHU ; Bei WANG ; Jiangbing SUN ; Shengqin HUANG
Chinese Journal of Biotechnology 2014;30(11):1742-1750
To use Agrobacterium rhizogenes-induced hairy roots to create new germplasm of Dianthus caryophyllus, we transformed D. caryophyllus with A. rhizogenes by leaf disc for plant regeneration from hairy roots. The white hairy roots could be induced from the basal surface of leaf explants of D. caryophyllus 12 days after inoculation with A. rhizogenes ATCC15834. The percentage of the rooting leaf explants was about 90% 21 days after inoculation. The hairy roots could grow rapidly and autonomously in liquid or solid phytohormone-free MS medium. The transformation was confirmed by PCR amplification of rol gene of Ri plasmid and silica gel thin-layer chromatography of opines from D. caryophyllus hairy roots. Hairy roots could form light green callus after cultured on MS+6-BA 1.0-3.0 mg/L + NAA 0.1-0.2 mg/L for 15 days. The optimum medium for adventitious shoots formation was MS + 6-BA 2.0 mg/L + NAA 0.02 mg/L, where the rate of adventitious shoot induction was 100% after cultured for 6 weeks. The mean number of adventitious shoot per callus was 30-40. The adventitious shoots can form roots when cultured on phytohormone-free 1/2 MS or 1/2 MS +0.5 mg/L NAA for 10 days. When the rooted plantlets transplanted in the substrate mixed with perlite sand and peat (volume ratio of 1:2), the survival rate was above 95%.
Agrobacterium
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Chromatography, Thin Layer
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Culture Media
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Dianthus
;
growth & development
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Plant Growth Regulators
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Plant Leaves
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Plant Roots
;
growth & development
;
Plants, Genetically Modified
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Rhizobium
;
Tissue Culture Techniques
;
Transformation, Genetic
8.Issues of large scale tissue culture of medicinal plant.
Dong-Mei LV ; Yuan YUAN ; Zhi-Lai ZHAN
China Journal of Chinese Materia Medica 2014;39(17):3413-3415
In order to increase the yield and quality of the medicinal plant and enhance the competitive power of industry of medicinal plant in our country, this paper analyzed the status, problem and countermeasure of the tissue culture of medicinal plant on large scale. Although the biotechnology is one of the most efficient and promising means in production of medicinal plant, it still has problems such as stability of the material, safety of the transgenic medicinal plant and optimization of cultured condition. Establishing perfect evaluation system according to the characteristic of the medicinal plant is the key measures to assure the sustainable development of the tissue culture of medicinal plant on large scale.
Drug Industry
;
methods
;
standards
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Medicine, Chinese Traditional
;
methods
;
standards
;
Plants, Genetically Modified
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Plants, Medicinal
;
genetics
;
growth & development
;
Quality Control
;
Tissue Culture Techniques
;
methods
;
standards
9.Induction of polyploid in hairy roots of Nicotiana tabacum and its plant regeneration.
Lili HOU ; Heping SHI ; Wu YU ; Po Keung Eric TSANG ; Cheuk Fai Stephen CHOW
Chinese Journal of Biotechnology 2014;30(4):581-594
By genetic transformation with Agrobacterum rhizogenes and artificial chromosome doubling techniques, we studied the induction of hairy roots and their polyploidization, and subsequent plant regeneration and nicotine determination to enhance the content of nicotine in Nicotiana tabacum. The results show that hairy roots could be induced from the basal surface of leaf explants of N. tabacum 8 days after inoculation with Agrobacterium rhizogenes ATCC15834. The percentage of the rooting leaf explants was 100% 15 days after inoculation. The hairy roots could grow rapidly and autonomously on solid or liquid phytohormones-free MS medium. The transformation was confirmed by PCR amplification of rol gene of Ri plasmid and paper electrophoresis of opines from N. tabacum hairy roots. The highest rate of polyploidy induction, more than 64.71%, was obtained after treatment of hairy roots with 0.1% colchicine for 36 h. The optimum medium for plant regeneration from polyploid hairy roots was MS+2.0 mg/L 6-BA +0.2 mg/L NAA. Compared with the control diploid plants, the hairy roots-regenerated plants had weak apical dominance, more axillary buds and more narrow leaves; whereas the polyploid hairy root-regenerated plants had thicker stems, shorter internodes and the colour, width and thickness of leaves were significantly higher than that of the control. Observation of the number of chromosomes in their root tip cells reveals that the obtained polyploid regenerated plants were tetraploidy, with 96 (4n = 96) chromosomes. Pot-grown experiments showed compared to the control, the flowering was delayed by 21 days in diploid hairy roots-regenerated plants and polyploid hairy root-regenerated plants. GC-MS detection shows that the content of nicotine in polyploid plants was about 6.90 and 4.57 times the control and the diploid hairy roots-regenerated plants, respectively.
Agrobacterium
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Plant Roots
;
growth & development
;
Plants, Genetically Modified
;
growth & development
;
Polyploidy
;
Regeneration
;
Tobacco
;
genetics
;
growth & development
;
Transformation, Genetic

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