1.Investigating the impact of silencing an RNA-binding protein gene SlRBP1 on tomato photosynthesis through RNA-sequencing analysis.
Xiwen ZHOU ; Liqun MA ; Hongliang ZHU
Chinese Journal of Biotechnology 2024;40(1):150-162
		                        		
		                        			
		                        			Photosynthesis in plants directly affects the synthesis and accumulation of organic matter, which directly influences crop yield. RNA-binding proteins (RBPs) are involved in the regulation of a variety of physiological functions in plants, while the functions of RBPs in photosynthesis have not been clearly elucidated. To investigate the effect of a glycine-rich RNA-binding protein (SlRBP1) in tomato on plant photosynthesis, a stably inherited SlRBP1 silenced plant in Alisa Craig was obtained by plant tissue culture using artificial small RNA interference. It turns out that the size of the tomato fruit was reduced and leaves significantly turned yellow. Chlorophyll(Chl) content measurement, Chl fluorescence imaging and chloroplast transmission electron microscopy revealed that the chloroplast morphology and structure of the leaves of tomato amiR-SlRBP1 silenced plants were disrupted, and the chlorophyll content was significantly reduced. Measurement of photosynthesis rate of wild-type and amiR-SlRBP1 silenced plants in the same period demonstrated that the photosynthetic rate of these plants was significantly reduced, and analysis of RNA-seq data indicated that silencing of SlRBP1 significantly reduced the expression of photosynthesis-related genes, such as PsaE, PsaL, and PsbY, and affected the yield of tomato fruits through photosynthesis.
		                        		
		                        		
		                        		
		                        			RNA
		                        			;
		                        		
		                        			Solanum lycopersicum/genetics*
		                        			;
		                        		
		                        			Photosynthesis/genetics*
		                        			;
		                        		
		                        			Chlorophyll
		                        			;
		                        		
		                        			RNA-Binding Proteins/genetics*
		                        			
		                        		
		                        	
2.Light-driven CO2 conversion system: construction, optimization and application.
Yamei GAN ; Liang GUO ; Cong GAO ; Wei SONG ; Jing WU ; Liming LIU ; Xiulai CHEN
Chinese Journal of Biotechnology 2023;39(6):2390-2409
		                        		
		                        			
		                        			The use of light energy to drive carbon dioxide (CO2) reduction for production of chemicals is of great significance for relieving environmental pressure and solving energy crisis. Photocapture, photoelectricity conversion and CO2 fixation are the key factors affecting the efficiency of photosynthesis, and thus also affect the efficiency of CO2 utilization. To solve the above problems, this review systematically summarizes the construction, optimization and application of light-driven hybrid system from the perspective of combining biochemistry and metabolic engineering. We introduce the latest research progress of light-driven CO2 reduction for biosynthesis of chemicals from three aspects: enzyme hybrid system, biological hybrid system and application of these hybrid system. In the aspect of enzyme hybrid system, many strategies were adopted such as improving enzyme catalytic activity and enhancing enzyme stability. In the aspect of biological hybrid system, many methods were used including enhancing biological light harvesting capacity, optimizing reducing power supply and improving energy regeneration. In terms of the applications, hybrid systems have been used in the production of one-carbon compounds, biofuels and biofoods. Finally, the future development direction of artificial photosynthetic system is prospected from the aspects of nanomaterials (including organic and inorganic materials) and biocatalysts (including enzymes and microorganisms).
		                        		
		                        		
		                        		
		                        			Carbon Dioxide/metabolism*
		                        			;
		                        		
		                        			Photosynthesis
		                        			;
		                        		
		                        			Metabolic Engineering
		                        			
		                        		
		                        	
3.Effects of stereoscopic traction on photosynthetic characteristics, yield, and quality of Codonopsis pilosula under organic cultivation.
Yang ZHOU ; Yuan CHEN ; Feng-Xia GUO ; Hong-Yan WANG ; Zi-Ping CAI
China Journal of Chinese Materia Medica 2023;48(10):2725-2731
		                        		
		                        			
		                        			To solve the serious problem of stem and leaf shading in the middle and late stage of traditional flat planting of Codonopsis pilosula, this study analyzed the effects of different stereoscopic traction heights on the photosynthetic characteristics and growth of C. pilosula and explored the optimal traction height to improve the yield and quality of C. pilosula. The experiment designed three stereo-scopic traction heights [H1(60 cm), H2(90 cm), and H3(120 cm)] with natural growth without traction as the control(CK). The results showed that the increase in stereoscopic traction heights broadened the growth space of stems and leaves of C. pilosula, enhanced the ventilation effect, significantly increased the average daily net photosynthetic rate of C. pilosula, promoted the absorption of intercellular CO_2, decreased the transpiration rate, and reduced the evaporation of water. Moreover, it effectively avoided the problem of weakened photosynthesis, maintained the carbon balance of individual plants, and promoted the growth and development of the C. pilosula roots. In terms of the seed yield of C. pilosula, it was ranked as H2>H1>H3>CK. To be specific, H1 increased by 213.41% compared with CK, H2 increased by 282.43% compared with CK, and H3 increased by 133.95% compared with CK. The yield and quality of C. pilosula were the highest in the H3 treatment group, with the fresh yield of 6 858.33 kg·hm~(-2), 50.59% higher than CK, dry yield of 2 398.33 kg·hm~(-2), 76.54% higher than CK, and lobetyolin content of 0.56 mg·g~(-1), 45.22% higher than CK. Therefore, the stereoscopic traction height has a great influence on the photosynthetic characteristics, yield, and quality of C. pilosula. Particularly, the yield and quality of C. pilosula can be optimized and improved in the traction height treatment of H3(120 cm). This planting method is worth popularizing and applying in the cultivated management of C. pilosula.
		                        		
		                        		
		                        		
		                        			Codonopsis
		                        			;
		                        		
		                        			Traction
		                        			;
		                        		
		                        			Photosynthesis
		                        			;
		                        		
		                        			Plant Leaves
		                        			;
		                        		
		                        			Plant Roots
		                        			
		                        		
		                        	
4.Physiological and transcriptional responses to heat stress in a typical phenotype of Pinellia ternata.
Jialu WANG ; Jialei CHEN ; Xiangyu ZHANG ; Xue FENG ; Xiwen LI
Chinese Journal of Natural Medicines (English Ed.) 2023;21(4):243-252
		                        		
		                        			
		                        			Pinellia ternata is an important medicinal plant, and its growth and development are easily threatened by high temperature. In this study, comprehensive research on physiological, cytological and transcriptional responses to different levels of heat stress were conducted on a typical phenotype of P. ternata. First, P. ternata exhibited tolerance to the increased temperature, which was supported by normal growing leaves, as well as decreased and sustained photosynthetic parameters. Severe stress aggravated the damages, and P. ternata displayed an obvious leaf senescence phenotype, with significantly increased SOD and POD activities (46% and 213%). In addition, mesophyll cells were seriously damaged, chloroplast thylakoid was fuzzy, grana lamellae and stroma lamellae were obviously broken, and grana thylakoids were stacked, resulting in a dramatically declined photosynthetic rate (74.6%). Moreover, a total of 16 808 genes were significantly differential expressed during this process, most of which were involved in photosynthesis, transmembrane transporter activity and plastid metabolism. The number of differentially expressed transcription factors in MYB and bHLH families was the largest, indicating that these genes might participate in heat stress response in P. ternata. These findings provide insight into the response to high temperature and facilitate the standardized cultivation of P. ternata.
		                        		
		                        		
		                        		
		                        			Pinellia/genetics*
		                        			;
		                        		
		                        			Heat-Shock Response/genetics*
		                        			;
		                        		
		                        			Photosynthesis/genetics*
		                        			;
		                        		
		                        			Plants, Medicinal/genetics*
		                        			;
		                        		
		                        			Phenotype
		                        			
		                        		
		                        	
5.Physiological regulation of salicylic acid on Helianthus tubeuosus upon copper stress and root FTIR analysis.
Jinxiang AI ; Jieke GE ; Ziyi ZHANG ; Wenqian CHEN ; Jiayi LIANG ; Xinyi WANG ; Qiaoyuan WU ; Jie YU ; Yitong YE ; Tianyi ZHOU ; Jinyi SU ; Wenwen LI ; Yuhuan WU ; Peng LIU
Chinese Journal of Biotechnology 2023;39(2):695-712
		                        		
		                        			
		                        			Phytoremediation plays an important role in the treatment of heavy metal pollution in soil. In order to elucidate the mechanism of salicylic acid (SA) on copper absorption, seedlings from Xuzhou (with strong Cu-tolerance) and Weifang Helianthus tuberosus cultivars (with weak Cu-tolerance) were selected for pot culture experiments. 1 mmol/L SA was sprayed upon 300 mg/kg soil copper stress, and the photosynthesis, leaf antioxidant system, several essential mineral nutrients and the changes of root upon copper stress were analyzed to explore the mechanism of copper resistance. The results showed that Pn, Tr, Gs and Ci upon copper stress decreased significantly compared to the control group. Meanwhile, chlorophyll a, chlorophyll b and carotenoid decreased with significant increase in initial fluorescence (F0), maximum photochemical quantum yield of PSⅡ (Fv/Fm), electron transfer rate (ETR) and photochemical quenching coefficient (qP) content all decreased. The ascorbic acid (AsA) content was decreased, the glutathione (GSH) value was increased, the superoxide dismutase (SOD), catalase (CAT) and ascorbate peroxidase (APX) activity in the leaves were decreased, and the peroxidase (POD) activity was significantly increased. SA increased the Cu content in the ground and root system, and weakened the nutrient uptake capacity of K, Ca, Mg, and Zn in the root stem and leaves. Spray of exogenous SA can maintain the opening of leaf stomata, improve the adverse effect of copper on photosynthetic pigment and PSⅡ reaction center. Mediating the SOD and APX activity started the AsA-GSH cycle process, effectively regulated the antioxidant enzyme system in chrysanthemum taro, significantly reduced the copper content of all parts of the plant, and improved the ion exchange capacity in the body. External SA increased the content of the negative electric group on the root by changing the proportion of components in the root, promoted the absorption of mineral nutrient elements and the accumulation of osmoregulatory substances, strengthened the fixation effect of the root on metal copper, and avoided its massive accumulation in the H. tuberosus body, so as to alleviate the inhibitory effect of copper on plant growth. The study revealed the physiological regulation of SA upon copper stress, and provided a theoretical basis for planting H. tuberosus to repair soil copper pollution.
		                        		
		                        		
		                        		
		                        			Antioxidants
		                        			;
		                        		
		                        			Copper
		                        			;
		                        		
		                        			Helianthus/metabolism*
		                        			;
		                        		
		                        			Salicylic Acid/pharmacology*
		                        			;
		                        		
		                        			Chlorophyll A/pharmacology*
		                        			;
		                        		
		                        			Spectroscopy, Fourier Transform Infrared
		                        			;
		                        		
		                        			Chlorophyll/pharmacology*
		                        			;
		                        		
		                        			Ascorbic Acid
		                        			;
		                        		
		                        			Superoxide Dismutase/metabolism*
		                        			;
		                        		
		                        			Photosynthesis
		                        			;
		                        		
		                        			Glutathione
		                        			;
		                        		
		                        			Plant Leaves
		                        			;
		                        		
		                        			Stress, Physiological
		                        			;
		                        		
		                        			Seedlings
		                        			
		                        		
		                        	
6.Advances in using adaptive laboratory evolution technology for engineering of photosynthetic cyanobacteria.
Jiawei GAO ; Xiaofei ZHU ; Tao SUN ; Lei CHEN ; Weiwen ZHANG
Chinese Journal of Biotechnology 2023;39(8):3075-3094
		                        		
		                        			
		                        			Cyanobacteria are the only prokaryotes capable of oxygenic photosynthesis, which have potential to serve as "autotrophic cell factories". However, the synthesis of biofuels and chemicals using cyanobacteria as chassis are suffered from poor stress tolerance and low yield, resulting in low economic feasibility for industrial production. Thus, it's urgent to construct new cyanobacterial chassis by means of synthetic biology. In recent years, adaptive laboratory evolution (ALE) has made great achievements in chassis engineering, including optimizing growth rate, increasing tolerance, enhancing substrate utilization and increasing product yield. ALE has also made some progress in improving the tolerance of cyanobacteria to high light intensity, heavy metal ions, high concentrations of salt and organic solvents. However, the engineering efficiency of ALE strategy in cyanobacteria is generally low, and the molecular mechanisms underpinning the tolerance to various stresses have not been fully elucidated. To this end, this review summarizes the ALE-associated technical strategies and their applications in cyanobacteria chassis engineering, following by discussing how to construct larger ALE mutation library, increase mutation frequency of strains and shorten evolution time. Moreover, exploration of the construction principles and strategies for constructing multi-stress tolerant cyanobacteria, and efficient analysis the mutant libraries of evolved strains as well as construction of strains with high yield and strong robustness are discussed, with the aim to facilitate the engineering of cyanobacteria chassis and the application of engineered cyanobacteria in the future.
		                        		
		                        		
		                        		
		                        			Technology
		                        			;
		                        		
		                        			Photosynthesis/genetics*
		                        			;
		                        		
		                        			Cyanobacteria/genetics*
		                        			;
		                        		
		                        			Light
		                        			;
		                        		
		                        			Biofuels
		                        			
		                        		
		                        	
7.Characterization the response of Chlamydomonas reinhardtii serine/threonine protein kinase mutant to blue light.
Wangning LI ; Mengjing LIANG ; Ze YANG ; Yanan LI ; Chunhui ZHANG ; Chunli JI ; Runzhi LI ; Song QIN ; Jinai XUE ; Hongli CUI
Chinese Journal of Biotechnology 2023;39(11):4563-4579
		                        		
		                        			
		                        			In order to investigate the molecular mechanism of silk/threonine protein kinase (STK)-mediated blue light response in the algal Chlamydomonas reinhardtii, phenotype identification and transcriptome analysis were conducted for C. reinhardtii STK mutant strain crstk11 (with an AphvIII box reverse insertion in stk11 gene coding region) under blue light stress. Phenotypic examination showed that under normal light (white light), there was a slight difference in growth and pigment contents between the wild-type strain CC5325 and the mutant strain crstk11. Blue light inhibited the growth and chlorophyll synthesis in crstk11 cells, but significantly promoted the accumulation of carotenoids in crstk11. Transcriptome analysis showed that 860 differential expression genes (DEG) (559 up-regulated and 301 down-regulated) were detected in mutant (STK4) vs. wild type (WT4) upon treatment under high intensity blue light for 4 days. After being treated under high intensity blue light for 8 days, a total of 1 088 DEGs (468 upregulated and 620 downregulated) were obtained in STK8 vs. WT8. KEGG enrichment analysis revealed that compared to CC5325, the crstk11 blue light responsive genes were mainly involved in catalytic activity of intracellular photosynthesis, carbon metabolism, and pigment synthesis. Among them, upregulated genes included psaA, psaB, and psaC, psbA, psbB, psbC, psbD, psbH, and L, petA, petB, and petD, as well as genes encoding ATP synthase α, β and c subunits. Downregulated genes included petF and petJ. The present study uncovered that the protein kinase CrSTK11 of C. reinhardtii may participate in the blue light response of algal cells by mediating photosynthesis as well as pigment and carbon metabolism, providing new knowledge for in-depth analysis of the mechanism of light stress resistance in the algae.
		                        		
		                        		
		                        		
		                        			Chlamydomonas reinhardtii/genetics*
		                        			;
		                        		
		                        			Photosynthesis/genetics*
		                        			;
		                        		
		                        			Plants/metabolism*
		                        			;
		                        		
		                        			Protein Kinases
		                        			;
		                        		
		                        			Threonine/metabolism*
		                        			;
		                        		
		                        			Carbon/metabolism*
		                        			;
		                        		
		                        			Serine/metabolism*
		                        			
		                        		
		                        	
8.Biological function and molecular mechanism of the transcription factor GLKs in plants: a review.
Shurong SHEN ; Junjie YUAN ; Yiling XU ; Bojun MA ; Xifeng CHEN
Chinese Journal of Biotechnology 2022;38(8):2700-2712
		                        		
		                        			
		                        			GLKs (GOLDEN 2-LIKEs) are a group of plant-specific transcription factors regulating the chloroplast biogenesis, differentiation and function maintains by triggering the expression of the photosynthesis-associated nuclear genes (PhANGs). The GLKs also play important roles in nutrient's accumulation in fruits, leaf senescence, immunity and abiotic stress response. The expression of GLK genes were affected by multiple hormones or environmental factors. Therefore, GLKs were considered as the key nodes of regulatory network in plant cells, and potential candidates to improve the photosynthetic capacity of crops. Since numerous researches of GLKs have been reported in plants, the biological function, molecular mechanism of GLKs genes and its applications in breeding were summarized and a GLK-mediated signaling network model was developed. This review may facilitate future research and application of GLKs.
		                        		
		                        		
		                        		
		                        			Chloroplasts/genetics*
		                        			;
		                        		
		                        			Gene Expression Regulation, Plant
		                        			;
		                        		
		                        			Photosynthesis/genetics*
		                        			;
		                        		
		                        			Plant Breeding
		                        			;
		                        		
		                        			Transcription Factors/metabolism*
		                        			
		                        		
		                        	
9.Transcriptional analysis of grape in response to weak light stress.
Tianchi CHEN ; Tao XU ; Xuefu LI ; Leyi SHEN ; Lingling HU ; Yanfei GUO ; Yonghong JIA ; Yueyan WU
Chinese Journal of Biotechnology 2022;38(10):3859-3877
		                        		
		                        			
		                        			Grape (Vitis vinifera L.) in production is frequently exposed to inadequate light, which significantly affects its agronomic traits via inhibiting their physiological, metabolic and developmental processes. To explore the mechanism how the grape plants respond to the weak light stress, we used 'Yinhong' grape and examined their physiology-biochemistry characteristics and transcriptional profile under different levels of weak light stress. The results showed that grape seedlings upon low intensity shading treatments were not significantly affected. As the shading stress intensity was strengthened, the epidermis cells, palisade tissue, and spongy tissue in the leaves were thinner, the intercellular space between the palisade tissue and spongy tissue was larger compared with that of the control, and the activities of superoxide dismutase, catalase and peroxidase were decreased gradually. Additionally, the soluble protein content increased and the free proline content decreased gradually. Compared with the control, significant changes in plant photosynthetic characteristics and physiology-biochemistry characteristics were observed under high intensity of shading (80%). RNA-seq data showed that the differentially expressed genes between CK and T2, CK and T4, T2 and T4 were 13 913, 13 293 and 14 943, respectively. Most of the enrichment pathways were closely related with the plant's response to stress. Several signaling pathways in response to stress-resistance, e.g. JA/MYC2 pathway and MAPK signal pathway, were activated under weak light stress. The expression level of a variety of genes related to antioxidation (such as polyphenol oxidase and thioredoxin), photosynthesis (such as phytochrome) was altered under weak light stress, indicating that 'Yinhong' grape may activate the antioxidation related pathways to cope with reactive oxygen species (ROS). In addition, it may activate the expression of photosynthetic pigment and light reaction structural protein to maintain the photosynthesis activity. This research may help better understand the relevant physiological response mechanism and facilitate cultivation of grape seedlings under weak light.
		                        		
		                        		
		                        		
		                        			Vitis/metabolism*
		                        			;
		                        		
		                        			Gene Expression Regulation, Plant
		                        			;
		                        		
		                        			Photosynthesis/genetics*
		                        			;
		                        		
		                        			Plant Leaves
		                        			;
		                        		
		                        			Light
		                        			;
		                        		
		                        			Seedlings/metabolism*
		                        			
		                        		
		                        	
10.Effects of shading on photosynthetic physiology and energy metabolism of Asarum forbesii.
Li-Xian LU ; Hong-Zhuan SHI ; Qiao-Sheng GUO ; Kun ZHAO ; Huai-Yang CHANG ; Jie ZOU ; Li-Yuan GUO ; Zhao-Rui YANG
China Journal of Chinese Materia Medica 2022;47(15):4048-4054
		                        		
		                        			
		                        			Light is the main source for plants to obtain energy.Asarum forbesii is a typical shade medicinal plant, which generally grows in the shady and wet place under the bushes or beside the ditches.It can grow and develop without too much light intensity.This experiment explores the effects of shading on the growth, physiological characteristics and energy metabolism of A.forbesii, which can provide reference and guidance for its artificial planting.In this experiment, A.forbesii was planted under 80%, 60%, 40%, 20% and no shade.During the vigorous growth period, the photosynthetic physiological characteristics such as fluorescence parameters, photosynthetic parameters, photosynthetic pigment content and ultrastructure, as well as the content of mitochondrial electron transport chain(ETC) synthase and nutrients were measured.The results showed that the photosynthetic pigment content, chlorophyll fluorescence parameters and net photosynthesis rate(P_n) decreased with the decrease of shading.Under 20%-40% shading treatment, the plants had damaged ultrastructure, expanded and disintegrated chloroplast, disordered stroma lamella and grana lamella, and increased osmiophi-lic granules and starch granules.The activities of nicotinamide adenine dinucleotide dehydrogenase(NADH), succinate dehydrogenase(SDH), cytochrome C oxidoreductase(CCO) and adenosine triphosphate(ATP) synthasewere positively related to light intensity.With the reduction of shading, the content of total sugar and protein in nutrients increased first and then decreased, and the content was the highest under 60% shade.In conclusion, under 60%-80% shading treatment, the chloroplast and mitochondria had more complete structure, faster energy metabolism, higher light energy-conversion efficiency, better absorption and utilization of light energy and more nutrient synthesis, which was more suitable for the growth and development of A.forbesii.
		                        		
		                        		
		                        		
		                        			Asarum
		                        			;
		                        		
		                        			Chlorophyll/metabolism*
		                        			;
		                        		
		                        			Chloroplasts
		                        			;
		                        		
		                        			Energy Metabolism
		                        			;
		                        		
		                        			Photosynthesis/physiology*
		                        			;
		                        		
		                        			Plant Leaves/metabolism*
		                        			
		                        		
		                        	
            
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