1.Advances in abscisic acid biosynthesis.
Kexin LI ; Ying WANG ; Mingdong YAO ; Wenhai XIAO
Chinese Journal of Biotechnology 2023;39(6):2190-2203
Abscisic acid, a plant hormone that inhibits growth, is a key factor in balancing plant endogenous hormones and regulating growth and metabolism. Abscisic acid can improve the drought resistance and salt tolerance of crops, reduce fruit browning, reduce the incidence rate of malaria and stimulate insulin secretion, so it has a broad application potential in agriculture and medicine. Compared with traditional plant extraction and chemical synthesis, abscisic acid synthesis by microorganisms is an economic and sustainable route. At present, a lot of progress has been made in the synthesis of abscisic acid by natural microorganisms such as Botrytis cinerea and Cercospora rosea, while the research on the synthesis of abscisic acid by engineered microorganisms is rarely reported. Saccharomyces cerevisiae, Yarrowia lipolytica and Escherichia coli are common hosts for heterologous synthesis of natural products due to their advantages of clear genetic background, easy operation and friendliness for industrial production. Therefore, the heterologous synthesis of abscisic acid by microorganisms is a more promising production method. The author reviews the research on the heterologous synthesis of abscisic acid by microorganisms from five aspects: selection of chassis cells, screening and expression enhancement of key enzymes, regulation of cofactors, enhancement of precursor supply and promotion of abscisic acid efflux. Finally, the future development direction of this field is prospected.
Abscisic Acid/metabolism*
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Plant Growth Regulators/metabolism*
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Plants/metabolism*
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Yarrowia/metabolism*
2.Advances on the microbial synthesis of plant-derived diterpenoids.
Yatian CHENG ; Hao TANG ; Lili SUN ; Yating HU ; Ying MA ; Juan GUO ; Luqi HUANG
Chinese Journal of Biotechnology 2023;39(6):2265-2283
Natural plant-derived diterpenoids are a class of compounds with diverse structures and functions. These compounds are widely used in pharmaceuticals, cosmetics and food additives industries because of their pharmacological properties such as anticancer, anti-inflammatory and antibacterial activities. In recent years, with the gradual discovery of functional genes in the biosynthetic pathway of plant-derived diterpenoids and the development of synthetic biotechnology, great efforts have been made to construct a variety of diterpenoid microbial cell factories through metabolic engineering and synthetic biology, resulting in gram-level production of many compounds. This article summarizes the construction of plant-derived diterpenoid microbial cell factories through synthetic biotechnology, followed by introducing the metabolic engineering strategies applied to improve plant-derived diterpenoids production, with the aim to provide a reference for the construction of high-yield plant-derived diterpenoid microbial cell factories and the industrial production of diterpenoids.
Diterpenes/metabolism*
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Biotechnology
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Metabolic Engineering
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Biosynthetic Pathways/genetics*
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Plants/genetics*
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Synthetic Biology
3.The role of plant WRKY transcription factors against salt stress: a review.
Xiangxiang YE ; Yongjiang BI ; Qiong RAN ; Xiaohui ZHANG ; Bangjun WANG
Chinese Journal of Biotechnology 2023;39(7):2600-2611
High salt content in soils severely hampers plant growth and crop yields. Many transcription factors in plants play important roles in responding to various stresses, but their molecular mechanisms remain unclear. WRKY transcription factors are one of the largest families of transcription factors in higher plants that are involved in and influence many aspects of plant growth and development. They play important roles in responding to salt stress. The regulation of gene expression by WRKY proteins is mainly achieved by binding to the DNA's specific cis-regulatory elements, the W-box elements (TTGACC). In recent years, there have been many studies revealing the roles and mechanisms of WRKY family members, from model plant Arabidopsis to agricultural crops. This paper reviews the latest research progress on WRKY transcription factors in response to salt stress and discusses the current challenges and future perspectives of WRKY transcription factor research.
Transcription Factors/metabolism*
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Plant Proteins/metabolism*
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Stress, Physiological/genetics*
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Salt Stress/genetics*
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Crops, Agricultural/genetics*
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Gene Expression Regulation, Plant
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Phylogeny
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Plants, Genetically Modified/genetics*
4.Soybean GmGolS2-2 improves drought resistance of transgenic tobacco.
Haiwei YU ; Shuang QIU ; Jun ZHANG ; Shanshan LI ; Tianguo SUN ; Tianyi MA ; Yan ZHAO ; Xu ZHAO ; Ying ZHAI
Chinese Journal of Biotechnology 2023;39(7):2762-2771
Galactinol synthase (GolS) genes play important roles in plant response to abiotic stress. In this research, the plant expression vector of soybean GmGolS2-2 gene was constructed and transformed into tobacco to study the drought tolerance of transgenic tobacco. A GmGolS2-2 gene with 975 bp coding sequence was cloned from soybean leaves by reverse transcription-polymerase chain reaction (RT-PCR). GmGolS2-2 was linked to the plant expression vector pRI101 by restriction enzyme sites Nde Ⅰ and EcoR Ⅰ, and transformed into tobacco by leaf disc method. Genomic DNA PCR and real-time PCR showed that three GmGolS2-2 transgenic tobacco plants were obtained. The growth status of GmGolS2-2 transgenic tobacco under drought stress was better than that of wild-type tobacco. After drought stress treatment, the electrolyte leakage and malondialdehyde content of transgenic tobacco were lower than those of wild-type tobacco, but the proline content and soluble sugar content were higher than those of wild-type tobacco. The results of real-time PCR showed that the heterologous expression of GmGolS2-2 increased the expression of stress-related genes NtERD10C and NtAQP1 in transgenic tobacco. The above results indicated that GmGolS2-2 improved drought resistance of transgenic tobacco.
Drought Resistance
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Tobacco/genetics*
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Soybeans/genetics*
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Plant Proteins/metabolism*
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Plants, Genetically Modified/genetics*
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Stress, Physiological/genetics*
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Droughts
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Gene Expression Regulation, Plant
5.Functional analysis on sucrose transporters in sweet potato.
Yiran LIU ; Zhengdan WU ; Weitai WU ; Chaobin YANG ; Cairui CHEN ; Kai ZHANG
Chinese Journal of Biotechnology 2023;39(7):2772-2793
Sweet potato is an important food crop that can also be used as an industrial raw material. Sucrose is the main form of long-distance carbohydrate transport in plants, and sucrose transporter (SUT) regulates the transmembrane transport and distribution of sucrose during plant growth and metabolism. Moreover, SUT plays a key role in phloem mediated source-to-sink sucrose transport and physiological activities, supplying sucrose for the sink tissues. In this study, the full-length cDNA sequences of IbSUT62788 and IbSUT81616 were obtained by rapid amplification of cDNA ends (RACE) cloning according to the transcripts of the two SUT coding genes which were differentially expressed in sweet potato storage roots with different starch properties. Phylogenetic analysis was performed to clarify the classification of IbSUT62788 and IbSUT81616. The subcellular localization of IbSUT62788 and IbSUT81616 was determined by transient expression in Nicotiana benthamiana. The function of IbSUT62788 and IbSUT81616 in sucrose and hexose absorption and transport was identified using yeast functional complementarity system. The expression pattern of IbSUT62788 and IbSUT81616 in sweet potato organs were analyzed by real-time fluorescence quantitative PCR (RT-qPCR). Arabidopsis plants heterologous expressing IbSUT62788 and IbSUT81616 genes were obtained using floral dip method. The differences in starch and sugar contents between transgenic and wild-type Arabidopsis were compared. The results showed IbSUT62788 and IbSUT81616 encoded SUT proteins with a length of 505 and 521 amino acids, respectively, and both proteins belonged to the SUT1 subfamily. IbSUT62788 and IbSUT81616 were located in the cell membrane and were able to transport sucrose, glucose and fructose in the yeast system. In addition, IbSUT62788 was also able to transport mannose. The expression of IbSUT62788 was higher in leaves, lateral branches and main stems, and the expression of IbSUT81616 was higher in lateral branches, stems and storage roots. After IbSUT62788 and IbSUT81616 were heterologously expressed in Arabidopsis, the plants grew normally, but the biomass increased. The heterologous expression of IbSUT62788 increased the soluble sugar content, leaf size and 1 000-seed weight of Arabidopsis plants. Heterologous expression of IbSUT81616 increased starch accumulation in leaves and root tips and 1 000-seed weight of seeds, but decreased soluble sugar content. The results obtained in this study showed that IbSUT62788 and IbSUT81616 might be important genes regulating sucrose and sugar content traits in sweet potato. They might carry out physiological functions on cell membrane, such as transmembrane transport of sucrose, sucrose into and out of sink tissue, as well as transport and unloading of sucrose into phloem. The changes in traits result from their heterologous expression in Arabidopsis indicates their potential in improving the yield of other plants or crops. The results obtained in this study provide important information for revealing the functions of IbSUT62788 and IbSUT81616 in starch and glucose metabolism and formation mechanism of important quality traits in sweet potato.
Ipomoea batatas/metabolism*
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Arabidopsis/metabolism*
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Sucrose/metabolism*
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Saccharomyces cerevisiae/metabolism*
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DNA, Complementary
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Phylogeny
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Plants, Genetically Modified/genetics*
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Membrane Transport Proteins/metabolism*
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Starch/metabolism*
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Plant Proteins/metabolism*
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Gene Expression Regulation, Plant
6.Blueberry attenuates liver injury in metabolic dysfunction-associated liver disease by promoting the expression of mitofilin/Mic60 in human hepatocytes and inhibiting the production of superoxide.
Ya REN ; Houmin FAN ; Lili ZHU ; Tao LIN ; Tingting REN
Chinese Journal of Cellular and Molecular Immunology 2023;39(4):318-324
Objective To study the effect and mechanism of blueberry on regulating the mitochondrial inner membrane protein mitofilin/Mic60 in an in vitro model of metabolic dysfunction-associated liver disease (MAFLD). Methods L02 human hepatocytes were induced by free fatty acids (FFA) to establish MAFLD cell model. A normal group, a model group, an 80 μg/mL blueberry treatment group, a Mic60 short hairpin RNA (Mic60 shRNA) transfection group, and Mic60 knockdown combined with an 80 μg/mL blueberry treatment group were established. The intracellular lipid deposition was observed by oil red O staining, and the effect of different concentrations of blueberry pulp on the survival rate of L02 cells treated with FFA was measured by MTT assay. The levels of alanine aminotransferase (ALT), aspartate aminotransferase (AST), triglyceride (TG), total cholesterol (TC), superoxide dismutase (SOD) activity, glutathione (GSH) and malondialdehyde (MDA) contents were measured by visible spectrophotometry. The expression of reactive oxygen species (ROS) in hepatocytes was observed by fluorescence microscopy, and the mRNA and protein expression of Mic60 were detected by real-time quantitative PCR and Western blot analysis, respectively. Results After 24 hours of FFA stimulation, a large number of red lipid droplets in the cytoplasm of L02 cells was observed, and the survival rate of L02 cells treated with 80 μg/mL blueberry was higher. The results of ALT, AST, TG, TC, MDA and the fluorescence intensity of ROS in blueberry treated group were lower than those in model group, while the levels of SOD, GSH, Mic60 mRNA and protein in blueberry treated group were higher than those in model group. Conclusion Blueberry promotes the expression of Mic60, increases the levels of SOD and GSH in hepatocytes, and reduces the production of ROS, thus alleviating the injury of MAFLD hepatocytes and regulating the disorder of lipid metabolism.
Humans
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Blueberry Plants/chemistry*
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Hepatocytes/metabolism*
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Liver/metabolism*
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Liver Diseases/metabolism*
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Reactive Oxygen Species/metabolism*
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Superoxide Dismutase/metabolism*
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Superoxides/metabolism*
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Mitochondrial Membranes/metabolism*
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Mitochondrial Proteins/metabolism*
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Plant Extracts/pharmacology*
7.Phenylpropanoid pathway in plants and its role in response to heavy metal stress: a review.
Wenjia GE ; Jianpan XIN ; Runan TIAN
Chinese Journal of Biotechnology 2023;39(2):425-445
Phenylpropanoid metabolic pathway is one of the most important secondary metabolic pathways in plants. It directly or indirectly plays an antioxidant role in plant resistance to heavy metal stress, and can improve the absorption and stress tolerance of plants to heavy metal ions. In this paper, the core reactions and key enzymes of the phenylpropanoid metabolic pathway were summarized, and the biosynthetic processes of key metabolites such as lignin, flavonoids and proanthocyanidins and relevant mechanisms were analyzed. Based on this, the mechanisms of key products of phenylpropanoid metabolic pathway in response to heavy metal stress were discussed. The perspectives on the involvement of phenylpropanoid metabolism in plant defense against heavy metal stress provides a theoretical basis for improving the phytoremediation efficiency of heavy metal polluted environment.
Plants/metabolism*
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Metals, Heavy/metabolism*
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Flavonoids/metabolism*
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Biodegradation, Environmental
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Antioxidants
8.Targeted innovative design of Bt Cry toxin insecticidal mimics.
Chongxin XU ; Yuan LIU ; Xiao ZHANG ; Xianjin LIU
Chinese Journal of Biotechnology 2023;39(2):446-458
Bt Cry toxin is the mostly studied and widely used biological insect resistance protein, which plays a leading role in the green control of agricultural pests worldwide. However, with the wide application of its preparations and transgenic insecticidal crops, the resistance to target pests and potential ecological risks induced by the drive are increasingly prominent and attracting much attention. The researchers seek to explore new insecticidal protein materials that can simulate the insecticidal function of Bt Cry toxin. This will help to escort the sustainable and healthy production of crops, and relieve the pressure of target pests' resistance to Bt Cry toxin to a certain extent. In recent years, the author's team has proposed that Ab2β anti-idiotype antibody has the property of mimicking antigen structure and function based on the "Immune network theory" of antibody. With the help of phage display antibody library and specific antibody high-throughput screening and identification technology, Bt Cry toxin antibody was designed as the coating target antigen, and a series of Ab2β anti-idiotype antibodies (namely Bt Cry toxin insecticidal mimics) were screened from the phage antibody library. Among them, the lethality of Bt Cry toxin insecticidal mimics with the strongest activity was close to 80% of the corresponding original Bt Cry toxin, showing great promise for the targeted design of Bt Cry toxin insecticidal mimics. This paper systematically summarized the theoretical basis, technical conditions, research status, and discussed the development trend of relevant technologies and how to promote the application of existing achievements, aiming to facilitate the research and development of green insect-resistant materials.
Insecticides/metabolism*
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Bacillus thuringiensis
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Endotoxins/pharmacology*
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Bacillus thuringiensis Toxins/metabolism*
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Hemolysin Proteins/pharmacology*
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Bacterial Proteins/chemistry*
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Plants, Genetically Modified/genetics*
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Pest Control, Biological
9.Genome-wide identification and characterization of the WOX gene family in Brassica juncea.
Wenlong YANG ; Chu XU ; Jiaqi HAN ; Xiaohui ZHANG ; Jiangping SONG ; Huixia JIA ; Haiping WANG
Chinese Journal of Biotechnology 2023;39(2):537-551
The WUSCHEL related-homeobox (WOX) family is one of the plant-specific transcription factor families, playing important roles in plant growth and development. In this study, 51 WOX gene family members were identified from the genome data of Brassica juncea by searching and screening with HUMMER, Smart and other software. Their protein molecular weight, amino acids numbers, and isoelectric point were analyzed by using Expasy online software. Furthermore, bioinformatics software was used to systematically analyze the evolutionary relationship, conservative region, and gene structure of the WOX gene family. The mustard WOX gene family was divided into three subfamilies: ancient clade, intermediate clade, and WUS clade/modern clade. Structural analysis showed that the type, organization form and gene structure of the conservative domain of WOX transcription factor family members in the same subfamily were highly consistent, while there was a certain diversity among different subfamilies. 51 WOX genes are distributed unevenly on 18 chromosomes of mustard. Most of the promoters of these genes contain cis acting elements related to light, hormone and abiotic stress. Using transcriptome data and real-time fluorescence quantitative PCR (qRT-PCR) analysis, it was found that the expression of mustard WOX gene was spatio-temporal specific, among which BjuWOX25, BjuWOX33, and BjuWOX49 might play an important role in the development of silique, and BjuWOX10, BjuWOX32, and BjuWOX11, BjuWOX23 respectively might play an important role in the response to drought and high temperature stresses. The above results may facilitate the functional study of mustard WOX gene family.
Mustard Plant/genetics*
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Multigene Family/genetics*
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Transcription Factors/metabolism*
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Plants/genetics*
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Promoter Regions, Genetic
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Phylogeny
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Gene Expression Regulation, Plant
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Plant Proteins/metabolism*
10.Effect of ACC oxidase gene AhACOs on salt tolerance of peanut.
Jianbin HUANG ; Wenjie ZHOU ; Lei FANG ; Mingming SUN ; Xin LI ; Jingjing LI ; Xiaoting LI ; Yanyan TANG ; Defeng JIANG ; Hong ZHU ; Jiongming SUI ; Lixian QIAO
Chinese Journal of Biotechnology 2023;39(2):603-613
ACC oxidase (ACO) is one of the key enzymes that catalyze the synthesis of ethylene. Ethylene is involved in salt stress response in plants, and salt stress seriously affects the yield of peanut. In this study, AhACO genes were cloned and their functions were investigated with the aim to explore the biological function of AhACOs in salt stress response, and to provide genetic resources for the breeding of salt-tolerant varieties of peanut. AhACO1 and AhACO2 were amplified from the cDNA of salt-tolerant peanut mutant M29, respectively, and cloned into the plant expression vector pCAMBIA super1300. The recombinant plasmid was transformed into Huayu22 by pollen tube injection mediated by Agrobacterium tumefaciens. After harvest, the small slice cotyledon was separated from the kernel, and the positive seeds were screened by PCR. The expression of AhACO genes was analyzed by qRT-PCR, and the ethylene release was detected by capillary column gas chromatography. Transgenic seeds were sowed and then irrigated with NaCl solution, and the phenotypic changes of 21-day-seedings were recorded. The results showed that the growth of transgenic plants were better than that of the control group Huayu 22 upon salt stress, and the relative content of chlorophyll SPAD value and net photosynthetic rate (Pn) of transgenic peanuts were higher than those of the control group. In addition, the ethylene production of AhACO1 and AhACO2 transgenic plants were 2.79 and 1.87 times higher than that of control peanut, respectively. These results showed that AhACO1 and AhACO2 could significantly improve the salt stress tolerance of transgenic peanut.
Salt Tolerance/genetics*
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Arachis/genetics*
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Plant Breeding
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Ethylenes/metabolism*
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Plants, Genetically Modified/genetics*
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Gene Expression Regulation, Plant
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Plant Proteins/genetics*

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