Molecular dissection of an intronic enhancer governing cold-induced expression of the vacuolar invertase gene in potato.

Potato (Solanum tuberosum) is the third most important food crop in the world. Potato tubers must be stored at cold temperatures to minimize sprouting and losses due to disease. However, cold temperatures strongly induce the expression of the potato vacuolar invertase gene (VInv) and cause reducing sugar accumulation. This process, referred to as "cold-induced sweetening," is a major postharvest problem for the potato industry. We discovered that the cold-induced expression of VInv is controlled by a 200 bp enhancer, VInvIn2En, located in its second intron. We identified several DNA motifs in VInvIn2En that bind transcription factors involved in the plant cold stress response. Mutation of these DNA motifs abolished VInvIn2En function as a transcriptional enhancer. We developed VInvIn2En deletion lines in both diploid and tetraploid potato using clustered regularly interspaced short palindromic repeat (CRISPR)/CRISPR-associated nuclease 9 (Cas9)-mediated gene editing. VInv transcription in cold-stored tubers was significantly reduced in the deletion lines. Interestingly, the VInvIn2En sequence is highly conserved among distantly related Solanum species, including tomato (Solanum lycopersicum) and other non-tuber-bearing species. We conclude that the VInv gene and the VInvIn2En enhancer have adopted distinct roles in the cold stress response in tubers of tuber-bearing Solanum species.

Drought priming at seedling stage improves photosynthetic performance and yield of potato exposed to a short-term drought stress.

Potato (Solanum tuberosum L.) is an important food and vegetable crop worldwide. In recent years, the arid environment resulting from climate change has caused a sharp decline in potato yield. To clarify the effect of drought priming at the seedling stage on the tolerance of potato plants to drought stress during tuber expansion, we conducted a pot experiment to investigate the physiological response of the plants generated from seed potatoes of the variety 'Favorita' to varied water supply conditions: normal water supply at the seedling stage (control), normal water supply at the seedling stage and drought stress at the mid-tuber-expansion stage (non-primed), and drought priming at the seedling stage plus drought stress at the mid-tuber-expansion stage (primed). Drought priming resulted in an increase in the number of small vascular bundles in potato plants compared to non-primed plants. It also altered the shape and density of stomata, enhancing water use efficiency and reducing whole-plant transpiration. The primed plants maintained the basal stem cambium for a longer time under drought stress, which gained an extended differentiation ability to generate a greater number of small vascular bundles compared to non-primed plants. Drought priming increased the amount and rate of dry matter translocation, and so reduced the adverse effects on tubers of potato under drought stress. Therefore, drought priming at the seedling stage improved the photosynthetic performance and yield, and probably enhanced the drought tolerance of potato.

Systematic Analysis of Galactinol Synthase and Raffinose Synthase Gene Families in Potato and Their Expression Patterns in Development and Abiotic Stress Responses.

Raffinose family oligosaccharides (RFOs) are very important for plant growth, development, and abiotic stress tolerance. Galactinol synthase (GolS) and raffinose synthase (RFS) are critical enzymes involved in RFO biosynthesis. However, the whole-genome identification and stress responses of their coding genes in potato remain unexplored. In this study, four StGolS and nine StRFS genes were identified and classified into three and five subgroups, respectively. Remarkably, a total of two StGolS and four StRFS genes in potato were identified to form collinear pairs with those in both Arabidopsis and tomato, respectively. Subsequent analysis revealed that StGolS4 exhibited significantly high expression levels in transport-related tissues, PEG-6000, and ABA treatments, with remarkable upregulation under salt stress. Additionally, StRFS5 showed similar responses to StGolS4, but StRFS4 and StRFS8 gene expression increased significantly under salt treatment and decreased in PEG-6000 and ABA treatments. Overall, these results lay a foundation for further research on the functional characteristics and molecular mechanisms of these two gene families in response to ABA, salt, and drought stresses, and provide a theoretical foundation and new gene resources for the abiotic-stress-tolerant breeding of potato.

Transcriptome analysis reveals the proline metabolic pathway and its potential regulation TF-hub genes in salt-stressed potato.

Potato (Solanum tuberosum) is currently the third most important food crop in the world. However, the production of potato is seriously threatened by salt stress, which often occurs in the facility cultivation environment, and the mining of salt tolerance genes in potato remains to be further studied. In this study, test-tube plantlets of DM potato were treated with 200-mM NaCl to simulate salt stress, and 15 cDNA libraries were constructed for RNA-seq analysis. A total of 8383 DEGs were identified, of which 3961 DEGs were shared among all the salt treatments, and 264 (7.15%) TF-coding genes were identified from these shared DEGs. KEGG enrichment analysis showed that most DEGs identified from the "arginine and proline metabolism" (ko00330) were enriched in the proline metabolic pathway, and their functions almost covered the whole proline metabolic process. Further analysis showed that expression levels of all the 13 structural DEGs in the pathway were significantly up-regulated and proline accumulation was also significantly increased under salt stress, and 13 TF-hub genes were discovered by WGCNA in the lightcyan and tan modules which were highly positively correlated with the proline contents. Correlation analysis revealed that the four TF-hub genes of the lightcyan module and seven structural DEGs of the proline metabolic pathway might be the potential candidate genes, especially the potential and novel regulatory gene StGLK014720. Furthermore, the dual-luciferase reporter assay confirmed that the key protein StGLK014720 could activate the promoters of both structural genes StAST021010 and StAST017480. In conclusion, these results lay the foundation for further study on the salt tolerance mechanism of potato, and provide a theoretical basis and new genetic resources for salt tolerance breeding of potato.

Night warming at the vegetative stage improves pre-anthesis photosynthesis and plant productivity involved in grain yield of winter wheat.

We conducted pot experiments during the 2018-2020 growing seasons to study the effects of night warming at different growth stages of wheat on the photosynthetic performance; accumulation, transportation, and distribution of dry matter; and grain yield of winter wheat. Night warming at all the different growth stages resulted in an elevation of wheat yield by increasing the 1000-grain weight and the number of grains per ear. Night warming during the period from jointing to booting stage resulted in the greatest increase in wheat yield. It also increased the amount of overall dry matter and transferrable amount of dry matter in plants and increased the distribution of dry matter to grains to increase grain weight. Night warming treatments at three different growth stages enhanced pre-anthesis photosynthetic capacity by increasing flag leaf net photosynthetic rate, chlorophyll content, and photochemical efficiency of winter wheat at the early stage of grain filling, especially in the night warming treatment from jointing to booting stage. Night warming not only increased the stomatal density and stomatal index of wheat leaves but also increased stomatal conductance and transpiration rate in the early stage of grain filling, thus being conducive to the smooth progress of photosynthesis. In conclusion, night warming treatment at different growth stages increased the photosynthesis of flag leaves at the early stage of grain filling, and promoted the accumulation of dry matter in plants after anthesis, which was conducive to the grain yield of winter wheat.

Night-Warming Priming at the Vegetative Stage Alleviates Damage to the Flag Leaf Caused by Post-anthesis Warming in Winter Wheat (Triticum aestivum L.).

The asymmetric warming in diurnal and seasonal temperature patterns plays an important role in crop distribution and productivity. Asymmetric warming during the early growth periods of winter wheat profoundly affects its vegetative growth and post-anthesis grain productivity. Field experiments were conducted on winter wheat to explore the impact of night warming treatment in winter (Winter warming treatment, WT) or spring (Spring warming treatment, ST) on the senescence of flag leaves and yield of wheat plants later treated with night warming during grain filling (Warming treatment during grain filling, FT). The results showed that FT decreased wheat yield by reducing the number of grains per panicle and per 1,000-grain weight and that the yield of wheat plants treated with FT declined to a greater extent than that of wheat plants treated with WT + FT or ST + FT. The net photosynthetic rate, chlorophyll content, and chlorophyll fluorescence parameters of the flag leaves of wheat plants treated with WT + FT or ST + FT were higher than those under the control treatment from 0 to 7 days after anthesis (DAA) but were lower than those under the control treatment and higher than those of wheat plants treated with FT alone from 14 to 28 DAA. The soluble protein and Rubisco contents in the flag leaves of wheat plants treated with WT + FT or ST + FT were high in the early grain-filling period and then gradually decreased to below those of the control treatment. These contents were greater in wheat plants treated with WT + FT than in wheat plants treated with ST + FT from 0 to 14 DAA, whereas the opposite was true from 21 to 28 DAA. Furthermore, WT + FT and ST + FT inhibited membrane lipid peroxidation by increasing superoxide dismutase and peroxidase activities and lowering phospholipase D (PLD), phosphatidic acid (PA), lipoxygenase (LOX), and free fatty acid levels in the early grain-filling period, but their inhibitory effects on membrane lipid peroxidation gradually weakened during the late grain-filling period. Night-warming priming alleviated the adverse effect of post-anthesis warming on yield by delaying the post-anthesis senescence of flag leaves.