Exogenous melatonin improves the chilling tolerance and preharvest fruit shelf life in eggplant by affecting ROS-and senescence-related processes

Low temperature is the most common abiotic stress factors during the eggplant cultivation in solar greenhouses.Melatonin plays important roles in plant resistance to low temperature.However,the role of melatonin in regulating chilling tolerance and extending the preharvest shelf life of eggplant fruits is still unknown.In this study,we investigated the effects of exogenous melatonin on eggplant plants and fruits in response to low temperature.Under simulated low-temperature conditions,exogenous melatonin significantly relieved the chilling symptoms of seedlings by reducing reactive oxygen species (ROS) and malondialdehyde (MDA) levels and relative leakage rates.These reductions were caused by higher superoxide dismutase (SOD) and catalase (CAT) activities and increased endogenous polyamine and melatonin levels compared with those in untreated seedlings.Notably,the expression levels of SOD,CAT1/2,and polyamine synthesis genes (ADC and ODC) were also increased by 100μmol·L~(-1)melatonin,as well as those of genes involved in melatonin synthesis (TDC,T5H,SNAT,ASMT,and COMT) and cold regulation (COR1,CBFa/b,and ZAT2/6/12).To further investigate the effects of melatonin on eggplant leaves and fruits under natural low temperature conditions,100μmol·L~(-1)melatonin was sprayed on the functional leaves at three days before commodity maturation.Melatonin significantly alleviated chilling injury in the leaves and pericarp and extended the preharvest shelf life of the fruit by increasing the expression of COR1,CBF,ZAT2/6/12,and API5 and decreasing the expression of senescence-related genes (NCED1/2 and SAG12).Therefore,100μmol·L~(-1)melatonin improved chilling tolerance and fruit shelf life by upregulating ZAT2/6/12 to affect ROS-and senescence-related processes,which provides a reference for alleviating cold stress and extending the preharvest fruit shelf life in eggplant.

Effect of Polyamine Priming on Chilling Tolerance of Lolium perenne during Seed Imbibition

[Objective] This study aimed to investigate the effect of potyamine priming on physiological and biochemical variations of Lolium perenne embryos and seed germination. [Method] With annual Lolium perenne （Diamond T and Grazing-8000） as experimental materials, after priming with 0.5 mmol/L putrescine （Put）, spermidine （Spd） and spermine （Spm） for 24 h and chilling imbibition at 5 ℃ for 12, 24, 36 and 48 h, the effect of Put, Spd and Spm priming on chilling tolerance and germination ability of annual Lolium perenne seeds during imbibition was investigated. [Result] Put, Spd and Spm priming improved the activities of peroxidase （POD）, catalase （CAT）, superoxide dismutase （SOD） and ascorbate peroxidase （APX） and content of soluble protein content under low temperature stress, significantly in-creased the germination rate, and shortened the average germination duration. After chilling imbibition for 48 h, compared with the control, the average germination rate of annual Lolium perenne seeds was improved by 15.5% and 12.0% after Put, Spd and Spm priming, and the average germination duration was shortened by 1.21 and 1.14 d. During seed imbibition, the chilling tolerance of Grazing-8000 was stronger than that of Diamond T. Overall, Put, Spd and Spm treatment could increase the chilling tolerance of annual Lolium perenne seeds during imbibition, and improve the germination ability of seeds under low temperature stress. [Conclusion] Results of this study provided theoretical basis for the application of seed priming technology in the production of annual ryegrass.

Analysis and Evaluation Indicator Selection of Chilling Tolerance of Different Cotton Genotypes

[Objectivc] This study aimed to investigate the chilling tolerance of seedlings of different cotton genotypes and screen appropriate indicators for assess- ing chilling tolerance, to establish reliable mathematical evaluation model for chilling tolerance of cotton, thus providing theoretical basis for breeding and promoting new chilling-tolerant cotton germplasms and large-scale evaluation of chilling tolerance of cotton varieties. [Method] Fifteen cotton varieties （lines） were used as experimental materials. The photosynthetic gas exchange parameters, chlorophyll fluorescence ki- netic parameters, chlorophyll content, relative soluble sugar content, malonaldehyde content, relative proiine content, relative conductivity and other 12 physiological indi- cators of seedling leaves under low temperature treatment （5 ℃, 12 h） and recovery treatment （25 ℃. 24 h） were determined; based on the chilling tolerance coefficient （CTC） of various individual indicators, the comprehensive evaluation of chilling toler- ance was conducled by using principal component analysis, hierarchical cluster anal- ysis and stepwise regression analysis. [Result] The results showed that the 12 indi- vidual physiological indicators could be classified into 7 independent comprehensive components by principal component analysis; 15 cotton varieties （lines） were clus- tered into three categories by using membership function method and hierarchical cluster analysis; the mathematical model for evaluating chilling tolerance of cotton seedlings was established： D =0.275 -0.244Fo1 ＋0.206Fv/Fm1＋0.326g,%-0.056SS ＋ 0.225MDA＋O.O38REC （FF=0.995）, and the evaluation accuracy of the equation was higher than 94.25%,0. Six identification indicators closely related to chilling tolerance were screened, including Fo,, Fv/Fm1, Seedling leaves of cotton varieties （lines） gs2, SS, MDA, and REC. [Conclusion] with high chilling tolerance are less dam- aged under low temperature stress, and are able to maintain relatively high photo- synthetic electron transport capacity and high stomatal conductance after recovery treatment, which is contributed to gas exchange and recovery of photosynthetic ca- pacity. Determination of the six indicators under the same stress condition can be adopted for rapid identification and prediction of the chilling tolerance of other cotton varieties, which provides basis for the breeding, promotion, identification and screen- ing of chilling tolerant germplasms.

Ectopic expression of antifreeze protein gene from Ammopiptanthus nanus confers chilling tolerance in maize

Improved chilling tolerance is important for maize production. Previous efforts in transgenics and marker-assisted breeding have not achieved practical results. In this study, the antifreeze protein(AnAFP) from the super-xerophyte Ammopiptanthus nanus was aligned to KnS-type dehydrins.Phosphorylation in vitro and subcellular localization showed that AnAFP was phosphorylated by maize casein kinase II and translocated from nucleus to cytoplasm under chilling stress. AnAFP also increased lactate dehydrogenase activity. A parent line of commercial maize hybrids was transformed with the AnAFP gene. Based on thermal asymmetric interlaced PCR, one hemizygous and two homozygous integration sites were identified in one T_(1) line. Ectopic expression of AnAFP in transgenic lines was confirmed by quantitative real-time PCR, RNA-seq, and Western blotting. After chilling treatment, the transgenic lines showed significantly improved phenotype, higher kernel production, survival rate and biomass, and lower relative electrolyte leakage and superoxide dismutation than the untransformed line. Thus, ectopic expression of AnAFP gene improved chilling tolerance in the transgenic lines, which could be used to apply for further safety assessment for commercial breeding.

Chilling Tolerance of Cucumber During Germination is Related to Expression of Lysine Decarboxylase Gene

Using cDNA-AFLP technique, a specific fragment was isolated from cucumber cultivar Changchun mici possessing chilling tolerance induced at low temperature （15℃）. This fragment, named cctr 132, could not be induced in the chilling sensitive cucumber cultivar Beijing jietou. After recovering the fragment, sequencing and translating, the results of blastx and blastp in GenBank of NCBI indicated that CCTR132 had 88.37% identities and 100% positives with Oryza sativa putative lysine decarboxylase-like protein respectively, and PGGXGTXXE, the putative conserved domain of lysine decarboxylase family, was detected from CCTR132, suggesting the cucumber chilling tolerance during germination is related to the expression of the lysine decarboxylase gene.

Heterologous expression of Lolium perenne antifreeze protein confers chilling tolerance in tomato

Antifreeze proteins（AFP） are produced by certain plants, animals, fungi and bacteria that enable them to survive upon extremely low temperature. Perennial rye grass, Lolium perenne, was reported to possess AFP which protects them from cold environments. In the present investigation, we isolated AFP gene from L. perenne and expressed it in tomato plants to elucidate its role upon chilling stress. The T1 transgenic tomato lines were selected and subjected to molecular, biochemical and physiological analyses. Stable integration and transcription of Lp AFP in transgenic tomato plants was confirmed by Southern blot hybridization and RT-PCR, respectively. Physiological analyses under chilling conditions showed that the chilling stress induced physiological damage in wild type（WT） plants, while the transgenic plants remained healthy. Total sugar content increased gradually in both WT and transgenic plants throughout the chilling treatment. Interestingly, transgenic plants exhibited remarkable alterations in terms of relative water content（RWC） and electrolyte leakage index（ELI） than those of WT. RWC increased significantly by 3-fold and the electrolyte leakage was reduced by 2.6-fold in transgenic plants comparing with WT. Overall, this report proved that Lp AFP gene confers chilling tolerance in transgenic tomato plants and it could be a potential candidate to extrapolate the chilling tolerance on other chilling-sensitive food crops.

The Relationship Between Heat-induced Chilling Tolerance and Endogenous ABA Levels in Mangoes

Heating 'Zhihua' mangoes (Mangifera indica L. cv. Zhihua) in hot air at 38℃ for 12hours significantly reduced the development of chilling injury following storage for 12days at 2t. Prolonging the heating up to 72hours acquired similar results. During the 12 days' storage at 2℃ following heating for 72hours, the membrane leakage of the mango peels showed little change, while it increased with time for the control fruits, and the endogenous ABA levels in the heated fruits were higher than that in the control. These results indicated that higher ABA rates were linked with stronger chilling tolerance in mangoes.

Chilling Tolerance and Physiological Parameters as Influenced by Grafting in Watermelon Seedlings

The influences of different rootstocks on chilling tolerance and physiological parameters in watermelon seedlings have been studied. The results showed that grafting improved the chilling tolerance. Compared with own-rooted watermelon seedlings, the grafted watermelon seedlings had lower chilling injury index, lower electrolytic leakage (%), lower malondialdehyde (MDA) content, higher chlorophyll and proline content, and higher activities of superoxide dismutase (SOD), ascorbate peroxidase (AsA-POD) and dehydroascorbate reductase (DR) in the leaves under low temperature stress. There was a considerable difference of chilling tolerance among different grafted watermelon seedlings due to the difference of rootstock chilling tolerance. After low temperature treatment, the grafted seedling with higher chilling tolerance had lower electrolytic leakage (%),lower MDA content, higher proline content and higher activities of SOD, AsA-POD and DR in the leaves compared with the grafted seedling with weaker chilling tolerance. From these, we could conclude that chilling tolerance of watermelon seedlings may be related to higher antioxidative ability and membrane stability in the plants. The chilling tolerance of grafted seedling could be properly evaluated by comprehensive physiological indexes but not a single physiological index.

Effect of LaCl_3 on the chilling tolerance of rice seedlings

The rice variety Tesanai 2 is susceptible tochilling.Exposure of the seedlings grown at 28±1℃ and under a photo flux density(PFD)of30 μmol/m~2s to 1℃ and under a PFD of 150μmol/m~2s for 2 d caused a physiological disor-der called chilling injury which reduced the sur-vival rate to 50%.The experimental results in-

Transcriptional activation and phosphorylation of OsCNGC9 confer enhanced chilling tolerance in rice

Low temperature is a major environmental factor that limits plant growth and productivity.Although transient elevation of cytoplasmic calcium has long been recognized as a critical signal for plant cold tolerance,the calcium channels responsible for this process have remained largely elusive.Here we report that OsCNGC9,a cyclic nucleotide-gated channel,positively regulates chilling tolerance by mediating cytoplasmic calcium elevation in rice(Oryza sativa).We showed that the loss-of-function mutant of OsCNGC9 is defective in cold-induced calcium influx and more sensitive to prolonged cold treatment,whereas OsCNGC9 overexpression confers enhanced cold tolerance.Mechanistically,we demonstrated that in response to chilling stress,OsSAPK8,a homolog of Arabidopsis thaliana OST1,phosphorylates and activates OsCNGC9 to trigger Ca2+influx.Moreover,we found that the transcription of OsCNGC9 is activated by a rice dehydration-responsive element-binding transcription factor,OsDREB1A.Taken together,our results suggest that OsCNGC9 enhances chilling tolerance in rice through regulating cold-induced calcium influx and cytoplasmic calcium elevation.

Genome-wide Association Analysis of Ten Chilling Tolerance Indices at the Germination and Seedling Stages in Maize

Maize seedlings are very sensitive to chilling, especially during the transition phase from heterotrophic to autotrophic growth. Genetic dissection of the genetic basis of chilling tolerance would provide useful information for genetic improvement of maize inbreds. In this study, genome-wide association analysis was conducted to explore the genetic architecture of maize chilling tolerance at the seed germination and seedling stages with an association panel of 125 inbreds. Ten tolerance indices （ratios of the performance of 10 germination rates and seedling growth-related traits under chilling stress and control conditions） were investigated to assess the ability of chilling tolerance of the inbreds, and a total of 43 single nucleotide polymorphisms associated with chilling tolerance were detected, with none of them being related to chilling tolerance at both the germination and seedling stages simultaneously. Correlation analysis also revealed that the genetic basis of chilling tolerance at the seed germination stage is generally different from that at the seedling stage. In addition, a total of 40 candidate genes involving 31 of the 43 single nucleotide polymorphisms were predicted, and were grouped into five categories according to their functions. The possible roles of these candidate genes in chilling tolerance were also discussed.

Protection of ultrastructure in chilling-stressed banana leaves by salicylic acid

Objective： Chilling tolerance of salicylic acid （SA） in banana seedlings （Musa acuminata cv., Williams 8818） was investigated by changes in ultrastructure in this study. Methods： Light and electron microscope observation. Results： Pretreatment with 0.5 mmol/L SA under normal growth conditions （30/22 ℃） by foliar spray and root irrigation resulted in many changes in ultrastructure of banana cells, such as cells separation from palisade parenchymas, the appearance of crevices in cell walls, the swelling of grana and stromal thylakoids, and a reduction in the number of starch granules. These results implied that SA treatment at 30/22 ℃ could be a type of stress. During 3 d of exposure to 7 ℃ chilling stress under low light, however, cell ultrastructure of SA-pretreated banana seedlings showed less deterioration than those of control seedlings （distilled water-pretreated）. Conclusion： SA could provide some protection for cell structure of chilling-stressed banana seedling.

Nitric oxide induced by polyamines involves antioxidant systems against chilling stress in tomato (Lycopersicon esculentum Mill.) seedling

Polyamines（PAs） and nitric oxide（NO） are vital signals in modulating plant response to abiotic stress. However, to our knowledge, studies on the relationship between NO and PAs in response to cold stress in tomato are limited. Accordingly, in this study, we investigated the effects of putrescine（Put） and spermidine（Spd） on NO generation and the function of Spd-induced NO in the tolerance of tomato seedling under chilling stress. Spd increased NO release via the nitric oxide synthase（NOS）-like and nitrate reductase（NR） enzymatic pathways in the seedlings, whereas Put had no such effect. Moreover, H2O2 might act as an upstream signal to stimulate NO production. Both exogenous NO donor（sodium nitroprusside（SNP）） and Spd enhanced chilling tolerance in tomato, thereby protecting the photosynthetic system from damage. Compared to chilling treatment alone, Spd enhanced the gene expressions of superoxide dismutase（SOD）, peroxidase（POD）, catalase（CAT）, and ascorbate peroxidase（APX）, and their enzyme activities in tomato leaves. However, a scavenger or inhibitor of NO abolished Spd-induced chilling tolerance and blocked the increased expression and activity due to Spd of these antioxidant enzymes in tomato leaves under chilling stress. The results showed that NO induced by Spd plays a crucial role in tomato＇s response to chilling stress.