Zhi-Feng Chen’s research while affiliated with Shanxi Agricultural University and other places

Bolting and flowering of vegetables are induced by vernalization in their early growth stage. This phenomenon is called premature bolting, and it has caused massive losses in production of vegetables such as cabbage, celery, carrot, radish, and spinach, etc. This review aimed to summarize studies on bolting and flowering pathways, physiological and biochemical changes, and underlined molecular mechanisms of various vegetable crop bolting involving genome and transcriptome analysis, and its association with vegetable breeding. This review could provide basic knowledge to carry out research on vegetable genetics and breeding and vegetable cultivation.

4-Coumarate: coenzyme A ligase (4CL; EC 6.2.1.12) is an important enzyme in the phenylpropanoid metabolic pathway that controls the biosynthesis of lignin and flavonoids. In this study, to identify the function of the Ag4CL3 gene of celery, the Ag4CL3 gene was cloned from celery cv. “Nanxuan Liuhe Ziqin”. Sequence analysis results showed that the Ag4CL3 gene contained an open reading frame (ORF) with a length of 1688 bp, and 555 amino acids were encoded. The Ag4CL3 protein was highly conserved among different plant species. Phylogenetic analysis demonstrated that the 4CL proteins from celery and carrot belonged to the same clade. The Ag4CL3 protein was mainly composed of 31.89% α-helixes, 18.02% extended strands, 6.67% β-turns, and 43.42% random coils, and the signal peptide was unfound. A total of 62 phosphorylation sites and a class-I superfamily of adenylate-forming domains were found. As the growth time increased, the plant height and stem thickness also increased, and the petiole lignin content increased and became lignified gradually. The relative expression levels of the Ag4CL3 gene in “Nanxuan Liuhe Ziqin” petioles were higher than those in other tissues, with the highest level occurring 70 d after sowing. The lignin contents in the transgenic Arabidopsis thaliana lines hosting the Ag4CL3 gene were higher than those in the WT. In this study, the overexpression of Ag4CL3 led to the significant upregulation of lignin biosynthesis gene expression in transgenic A. thaliana plants, except for AtPAL, AtCCR, and AtLAC. This study speculates that Ag4CL3 genes are related to lignin synthesis in A. graveolens.

Solar greenhouses are important in the vegetable production and widely used for the counter-season production in the world. However, the CO2 consumed by crops for photosynthesis after sunrise is not supplemented and becomes chronically deficient due to the airtight structure of solar greenhouses. Vegetable crops cannot effectively utilize light resources under low-CO2 environment, and this incapability results in reduced photosynthetic efficiency and crop yield. We used cucumber as a model plant and generated several sets of transgenic cucumber plants overexpressing individual genes, including β-carbonic anhydrase 1 (CsβCA1), β-carbonic anhydrase 4 (CsβCA4), and sedoheptulose-1,7-bisphosphatase (CsSBP); fructose-1,6-bisphosphate aldolase (CsFBA), and CsβCA1 co-expressing plants; CsβCA4, CsSBP, and CsFBA co-expressing plants (14SF). The results showed that the overexpression of CsβCA1, CsβCA4, and 14SF exhibited higher photosynthetic and biomass yield in transgenic cucumber plants under low-CO2 environment. Further enhancements in photosynthesis and biomass yield were observed in 14SF transgenic plants under low-CO2 environment. The net photosynthesis biomass yield and photosynthetic rate increased by 49% and 79% compared with those of the WT. However, the transgenic cucumbers of overexpressing CsFBA and CsSBP showed insignificant differences in photosynthesis and biomass yield compared with the WT under low-CO2.environment. Photosynthesis, fluorescence parameters, and enzymatic measurements indicated that CsβCA1, CsβCA4, CsSBP, and CsFBA had cumulative effects in photosynthetic carbon assimilation under low-CO2 environment. Co-expression of this four genes (CsβCA1, CsβCA4, CsSBP, and CsFBA) can increase the carboxylation activity of RuBisCO and promote the regeneration of RuBP. As a result, the 14SF transgenic plants showed a higher net photosynthetic rate and biomass yield even under low-CO2environment.These findings demonstrate the possibility of cultivating crops with high photosynthetic efficiency by manipulating genes involved in the photosynthetic carbon assimilation metabolic pathway.

Steroidal glycoalkaloids (SGAs), the nitrogen-containing compounds produced primarily by Liliaceae and Solanaceae species, are toxic to animals and humans and have putative roles in defense against pests. UDP-glycosyltransferases (UGTs) catalyze the final glycosylation steps of SGA biosynthesis. Although previously published studies focused on the effect of UGT proteins on SGA biosynthesis, research to understand the effects of constitutive overexpression of UGTs on plant phenotype and fruit development is limited. The constitutive overexpression of a UGT encoding gene, GAME 17, may provide an alternative method to study the role of UGTs on the fruit development. In this study, we have identified 162 SlUGT proteins in tomato that are classified into t 23 groups. Gene structure and motif analyses have demonstrated that all SlUGTs have similar intron/exon distribution and motif compositions. RNA-seq data analysis has shown that SlUGTs exhibit differential expression patterns in different organs or different stages of fruit development. When the constitutive promoter 35S is used to control the expression of GAME 17, we have observed significant differences in growth parameters (i.e., plant height, leaf length, leaf width, internode length, and stem diameter) between WT and transgenic plants under high-CO2 conditions, and slight differences in growth parameters between WT and transgenic plants have beendetected. In addition, the contents of glucose, fructose, and soluble sugar of transgenic plants are significantly higher than those of WT plants. The increases in glucose, fructose, and soluble sugar in transgenic tomato fruits at three developmental stages under high-CO2 conditions are significantly higher than under natural conditions. This study provides additional evidence that the GAME 17 gene plays an important role in controlling plant phenotype and sugar homeostasis, especially in environments with high concentration of CO2.

Genes in the carbonic anhydrase (CA) family encode zinc metalloenzymes that catalyze the reversible interconversion of carbon dioxide and water to bicarbonate and protons. Although CAs play key roles in diverse biological processes involving carboxylation and decarboxylation, including photosynthesis and respiration, plant growth and response to stress, the characteristics of CA gene family members in tomato remain unclear. In this study, we performed an exhaustive search of the tomato genome and accordingly identified 14 CA genes that are unevenly distributed on the 12 tomato chromosomes. We examined in detail the structures, conserved motifs, phylogenetic relationships and duplications of these genes, and for functional characterization, also undertook RNA-seq analyses to assess the transcript levels of CA genes in various tissues and organs and at different developmental stages. Furthermore, we investigated the expression patterns of the CA genes in response to salt stress. We found that some family members exhibited tissue-specific expression, whereas others were more ubiquitously expressed. Our results will provide a valuable foundation for further studies on the CA genes in tomato and other plants in the Solanaceae family.

Carbon dioxide (CO2) is very important for photosynthesis of green plants. CO2 concentration in the atmosphere is relatively stable, but it drops sharply after sunrise due to the tightness of the greenhouse and the absorption of CO2 by vegetable crops. Vegetables in greenhouses are chronically CO2 starved. To investigate the feasibility of using genetic engineering to improve the photosynthesis and yield of greenhouse cucumber in a low CO2 environment, five genes encoding glyoxylate carboligase (GCL), tartronic semialdehyde reductase (TSR), and glycolate dehydrogenase (GlcDH) in the glycolate catabolic pathway of Escherichia coli were partially or completely introduced into cucumber chloroplast. Both partial pathway by introducing GlcDH and full pathway expressing lines exhibited higher photosynthetic efficiency and biomass yield than wild-type (WT) controls in low CO2 environments. Expression of partial pathway by introducing GlcDH increased net photosynthesis by 14.9% and biomass yield by 44.9%, whereas the expression of the full pathway increased seed yield by 33.4% and biomass yield by 59.0%. Photosynthesis, fluorescence parameters, and enzymatic measurements confirmed that the introduction of glycolate catabolic pathway increased the activity of photosynthetic carbon assimilation-related enzymes and reduced the activity of photorespiration-related enzymes in cucumber, thereby promoting the operation of Calvin cycle and resulting in higher net photosynthetic rate even in low CO2 environments. This increase shows an improvement in the efficiency of the operation of the photosynthetic loop. However, the utilization of cucumber of low concentration CO2 was not alleviated. This study demonstrated the feasibility of introducing the pathway of exogenous glycolate catabolic pathway to improve the photosynthetic and bio-yield of cucumber in a low CO2 environment. These findings are of great significance for high photosynthetic efficiency breeding of greenhouse cucumber.

Tomato yellow leaf curl virus (TYLCV) is a globally devastating disease that affects the production of many crops, particularly tomato, leading to considerable economic losses. RNA interference, which allows sequence-specific gene silencing at the post-transcriptional level, is an effective method to obtain virus-resistant lines of crops. Agrobacterium-mediated transient expression assay has emerged as a rapid and useful approach to evaluate gene functions in plants without the need to produce transgenic lines. In this study, we transformed the plant expression vector pBIN438-AV1-AC1-AC3(i/r), which consists of the inverted repeat of Delta AV1, Delta AC1, and Delta AC3 fusion fragments into Agrobacterium tumefaciens EHA105 to generate tomato lines with high viral resistance. The viral resistance of the fusion gene was evaluated by transient expression. The test plants did not show disease symptoms at 35 d postinoculation with TYLCV. This study provided an important method for plant antiviral breeding and safe tomato production.