Advances in Molecular Regulatory Mechanisms of Fruit Trees under Low Temperature Stress

The most recent research findings on the tolerance of fruit trees to cold stress are reviewed from a molecular perspective,including the perception and transduction of low temperature calcium signaling,CBF-dependent molecular regulatory mechanisms,non-CBF-dependent molecular regulatory mechanisms,and so forth.The objective is to provide a reference basis for further improving the cold resistance of fruit trees and cultivating new varieties of hardy plants.

Effects of Low Temperature Stress on Germination and Physiological Characteristics of Different Sweet Maize Varieties

[Objectives]The paper was to explore the effects of low temperature stress on germination and physiological characteristics of different sweet maize varieties.[Methods]Taking Taitian 264,Zhexuetian 1 and Chaotian 4 as the research objects,the changes in germination potential,germination index,plant height,biomass,and antioxidant enzyme activity of maize seeds were studied under optimal temperature conditions(25℃)and low temperature stress conditions(10℃).[Results]Under 10℃stress,the germination rate and germination index of Taitian 264 were higher than that of Zhexuetian 1 and Chaotian 4.Under low temperature stress,Taitian 264 exhibited the least reduction in height and biomass,while Zhexuetian 1 had the most reduction.Additionally,the SOD and POD activities of Taitian 264 were higher than that of Zhexuetian 1 and Chaotian 4 under both temperature conditions,while the MDA content of Taitian 264 was lower.Taitian 264 showed strong germination ability against low temperature stress.[Conclusions]This study provides a basis for timely sowing practices of sweet maize in agricultural production.

De Novo Transcriptome Analysis in Hevea brasiliensis to Unveil Genes Involved in Low Temperature Stress Response

Low temperature is one of the adversities threatening the growth and development and reduces the yield of rubber trees.However,molecular mechanisms toward rubber trees in response to low temperature are largely unclear.In this study,7,159 and 7,600 differentially expressed genes(DEGs)were identified in‘Reyan 73397’rubber trees.Through GO analysis,the catalytic activity was the representative of the GO term in the only DEGs at the two studied temperatures(room temperature and 4°C,respectively),while KEGG analysis showed that carbon metabolism was the most important grouping under the comparison of these two temperatures.In addition,expression of 9 members of transcription factor MYB family genes were further verified by qRT-PCR,and MYB family genes may play important roles in the regulation of rubber trees under low temperature stress.This study provided a theoretical foundation for(1)revealing the molecular mechanisms of rubber trees in response to low temperature and(2)breeding of tolerant varieties of rubber trees.

Analysis of microRNA Expression Characteristics Related to Low Temperature Stress in Chewing Cane

[Objectives]To explore microRNA expression characteristics related to low temperature stress in chewing cane.[Methods]The research on miRNA under abiotic stress of sugarcane at home and abroad mainly focused on the types and regulation of miRNA under cold,heat,drought,high salt,and mechanical stress.However,there are few studies on miRNA under low temperature stress in chewing cane.The target genes of miR394 and miR825 in chewing cane were predicted and functionally analyzed by bioinformatics technology.[Results]The results showed that the target genes of miR394 and miR825 were mainly members of the WRKY transcription factor family,involved in plant growth,development and stress resistance.Real-time fluorescence quantitative PCR analyzed the expression characteristics of target miRNA in different tissues of chewing cane at different periods of low temperature stress.[Conclusions]The results showed that the expression of chewing cane miR394 and miR408 had temporal and spatial specificity and tissue specificity,both of which could respond to low temperature stress with significant differential expression.

Gene Expression and Activities of SOD in Cucumber Seedlings Were Related with Concentrations of Mn^(2+),Cu^(2+),or Zn^(2+) Under Low Temperature Stress

Effects of increasing Mn^2＋, Cu^2＋, or Zn^2＋ on SOD expressions were studied in cucumber seedlings under low temperature stress. Both gene expressions and activities of Cu/Zn-SOD and Mn-SOD in cucumber seedling leaves were induced by increasing Mn^2＋, Cu^2＋, or Zn^2＋ under low temperature stress, especially 48 h afterwards. The activities of Cu/Zn-SOD and Mn-SOD at 0 and 48 h after treatment were in accordance with their gene expression levels, which implied that the transcriptional regulation plays key roles in regulating their activities at the early stage of low temperature stress. Gene expressions of Cu/Zn-SOD and Mn-SOD declined at 96 h, but Cu/Zn-SOD and Mn-SOD activities still remain high, which suggested that Cu/Zn-SOD and Mn-SOD activities might be regulated by other factors after transcription at the later stage of low temperature stress. Therefore, we concluded that the increasing Mn^2＋, Cu^2＋, or Zn^2＋ could increase the capacity of scavenging ROS in cucumber seedlings under low temperature stress by inducing gene expressions of Cu/ Zn-SOD and Mn-SOD, elevating activities of Cu/Zn-SOD, Mn-SOD, or regulating other factors after transcription.

Effects of Phosphate Fertilizer on Cold Tolerance and Its Related Physiological Parameters in Rice Under Low Temperature Stress

The study was designated to explore the physiological mechanism of cold tolerance enhanced by phosphate in rice. An experiment was conducted to investigate the effects of different levels of phosphate fertilizer on cold tolerance and its related physiological parameters in rice seedings （chilling-sensitive cv. Changbai 9 and chilling-tolerant cv. Jijing 81） under low temperature stress. At the same time, the identification of cold tolerance was conducted. Compared with the normal temperature treatment, the relative chlorophyll content, photosynthesis rate, Fv/Fm and qP decreased and index of unsaturated fatty acid increased in rice under low temperature stress. The effect of chilling-sensitive cultivars was more than that of chilling-tolerant cultivars, more phosphorus fertilizer properly improved seedling quality of rice, slowed relative chlorophyll content dropping degree of rice seeding, increased photosynthesis rate, Fv/Fm, qP and index of unsaturated fatty acids, and enhanced the ability to chilling-tolerant cultivars under low temperature. The effect on chilling-tolerant cultivars was significantly higher than that on chilling sensitive cultivars by applying more phosphorus fertilizer. Phosphate regulated photosynthetic physiology and membrane fluidity to reduce injury by low temperature, and increasd the cold tolerance capacity of rice.

Evaluation of Rice Seedling Tolerance to Constant and Intermittent Low Temperature Stress

M202, IR50 and advanced backcross （BC4F6） lines carrying the cold tolerance QTLs qCTS4 and qCTS12 were evaluated using three low temperature stress assays （constant, intermittent and constant with recovery period）. Under constant stress, two BC4F6 lines （MIb 4853-9 and MIb 6885-2） exhibited differences in seedling growth and accumulation of stress-related compounds although both carry the two QTLs in the IRS0 background. Differences between these lines were also observed in the constant with recovery period assay, although both of them performed comparably under intermittent stress, MIb 6885-2 performed more similarly to M202 in the constant and constant with recovery period assays, suggesting that this line contains an introgressed region（s） not present in MIb 4853-9. The three assays were also applied to assess variations in cold tolerance in a set of diverse germplasms. Performance metrics and visual ratings were comparable for evaluating tolerance to constant stress. However, differences in response to intermittent stress were more evident from growth and total chlorophyll measurements than from visual ratings. With regard to the constant with recovery period assay, about 15% of the germplasm accessions did not recover. However, some lines which exhibited chilling injuries of comparable severity were able to overcome them. This recovery phenotype may improve rice performance in the field and warrant further investigation.

Effects of Low Temperature Stress on Seedling Growth of Plateau Japonica Rice

[ Objective ] The study aimed to screen rational seedling raising date and seedling raising mode for plateau japonica rice, in order to reduce the effect of low temperature on seedlings. [ Method ] The seedlings of conventional japonica rice Hexi 22-2 and giant embryo No. 1 japonica rice were treated at day tempera- ture 12℃/night temperature 8℃ and day temperature 14 ℃/night temperature 10 ℃, and the indicators including chlorophyll content, soluble sugar content, sol- uble protein content, survival rate, plant height, number of ≥ 1 cm roots, leaf wilting degree and aboveground dry matter weight were determined after treated for 0, 3 and 6 d, respectively. [ Result] Chlorophyll content of seedlings decreased when treated by low temperature for a certain period; with the prolongation of treat- ment time, contents of soluble sugar and soluble protein in some low temperature treatments increased, while plant height, number of ≥ 1 cm roots and aboveground dry matter weight increased slowly, but the growth rate was significantly lower than that at room temperature; with the decrease of temperature and the prolongation of treatment time, leaf wilting degree increased. The comprehensive experimental results showed that with the increasing duration of low temperature, when two japonica rice varieties grew under the same low temperature, seedlings grew more and more slowly, probably because the lower the treatment temperature, the greater the damage on plants. Giant Embryo No. 1 had weaker cold tolerance than Hexi 22-2. [ Conclusion] The study provides a theoretical basis for formula- tion of seedling raising technique of plateau japonica rice.

Physiological Response and Transcriptome Analysis of Cotton Leaves under Low Temperature Stress at the Two-leaf Stage

[Objectives]To investigate the effect of low temperature treatment on cotton leaves at the two-leaf stage.[Methods]Two cotton varieties(CN01 and SJB016(low temperature-tolerant))were used as the trial materials.They were treated at 25(CK)and 12℃(low temperature)for 0,3,6,12,24,48 and 72 h,respectively.Then,the changes in the contents of MDA,SS and Pro in the cotton leaves were analyzed.Based on the analysis results,RNA-seq verification was performed.[Results]Two cotton varieties(CN01 and SJB016(low temperature-tolerant))were used as the trial materials.They were treated at 25(CK)and 12℃(low temperature)for 0,3,6,12,24,48 and 72 h,respectively.Then,the changes in the contents of MDA,SS and Pro in the cotton leaves were analyzed.Based on the analysis results,RNA-seq verification was performed.[Conclusions]These genes may play an important role in improving the cold resistance of cotton.

Effects of PQQ on Protective System in Cucumber Cotyledons under Low Temperature Stress

The authors have studied the effects of Pyrroioguinloine quinone (PQQ) on superoxide dismutase (SOD), Ascorbid acid(AsA) peroxidase (AsAPOD), glutathion (GSH) and electrolytic leakage of cotyledon in cucumber seedling under low temperature stress, meanwhile, 8-hydroquinone (8-HQ) and AsA (activeoxiygen scavengers) have been made use of in comparison with PQQ. The results indicate that the activities of SOD, AsAPOD and content of GSH can be increased by PQQ. The relative conductivity of cotyledon in cucumber seedling is decreased for PQQ possesses the ability of cleaning up free redical of oxygen. We came to the conclusion that PQQ can act as a kind of active oxygen scavenger and adjust the metabolism on free radical of oxygen to balance in plants and enhance resistance finally in plants.

Genome-Wide Exploration of the Grape GLR Gene Family and Differential Responses of VvGLR3.1 and VvGLR3.2 to Low Temperature and Salt Stress

Grapes,one of the oldest tree species globally,are rich in vitamins.However,environmental conditions such as low temperature and soil salinization significantly affect grape yield and quality.The glutamate receptor(GLR)family,comprising highly conserved ligand-gated ion channels,regulates plant growth and development in response to stress.In this study,11 members of the VvGLR gene family in grapes were identified using whole-genome sequence analysis.Bioinformatic methods were employed to analyze the basic physical and chemical properties,phylogenetic trees,conserved domains,motifs,expression patterns,and evolutionary relationships.Phylogenetic and collinear analyses revealed that the VvGLRs were divided into three subgroups,showing the high conservation of the grape GLR family.These members exhibited 2 glutamate receptor binding regions(GABAb and GluR)and 3-4 transmembrane regions(M1,M2,M3,and M4).Real-time quantitative PCR analysis demonstrated the sensitivity of all VvGLRs to low temperature and salt stress.Subsequent localization studies in Nicotiana tabacum verified that VvGLR3.1 and VvGLR3.2 proteins were located on the cell membrane and cell nucleus.Additionally,yeast transformation experiments confirmed the functionality of VvGLR3.1 and VvGLR3.2 in response to low temperature and salt stress.Thesefindings highlight the significant role of the GLR family,a highly conserved group of ion channels,in enhancing grape stress resistance.This study offers new insights into the grape GLR gene family,providing fundamental knowledge for further functional analysis and breeding of stress-resistant grapevines.

Regulation of Calcium on Peanut Photosynthesis Under Low Night Temperature Stress

The effects of different levels of CaCl 2 on photosynthesis under low night temperature（8°C） stress in peanuts were studied in order to find out the appropriate concentration of Ca2＋through the artificial climate chamber potted culture test.The results indicated that Ca2＋,by means of improving the stomatal conductivity of peanut leaves under low night temperature stress,may mitigate the decline of photosynthetic rate in the peanut leaves.The regulation with 15 mmol L-1CaCl 2（Ca15） was the most effective,compared with other treatments.Subsequently,the improvement of Ca2＋on peanut photosynthesis under low night temperature stress was validated further through spraying with Ca15,Ca2＋chelator（ethylene glycol bis（2-aminoethyl） tetraacetic acid; EGTA） and calmodulin antagonists（trifluonerazine; TFP）.And CaM（Ca2＋-modulin） played an important role in the nutritional signal transduction for Ca2＋mitigating photosynthesis limitations in peanuts under low night temperature stress.

Effects of Low Temperature on Physiological Responses of Two White Clover Cultivars

In order to explore the effects of low temperature on the two different genotypes of white clover, electrical conductivity （EC）, soluble sugar, malondialdehyde （MDA） and some other physiological indexes of plants were compared under laboratory conditions. Seedlings of Longping No.1 and Koala were undergone a series of degrading temperatures set in turn 25℃, 10℃, 5℃, 0℃, –5℃ and –10℃. The results showed that the activities of superoxide dismutase （SOD）, electrical conductivity, soluble sugar, free proline and MDA increased obviously with the decrease of stress temperature, while the content of starch decreased, and the content of soluble protein increased first and then decreased during the process. Comprehensive assessments on physiological responses to low temperature stress indicated that the cultivar Longping No.1 had stronger cold resistance than that of cultivar Koala.

Potentiality of Different Seed Priming Agents to Mitigate Cold Stress of Winter Rice Seedling

Seed priming has proved to be an effective pre-germination seed invigoration technique for different crops to improve seed and seedling performance under different abiotic stresses.In Bangladesh,winter rice is very often exposed to cold waves just after sowing in the nursery bed resulting in poor seed germination and seedling emergence,yellowish and thin seedlings production,and a very low survival rate.Seed priming may mitigate the cold stress during seed germination and seedling emergence and helps in the quality seedling production of winter rice.To evaluate the efficacy of different seed priming techniques in increasing seedling emergence,growth,vigor and survivability of winter rice cultivars under cold stress,a pot experiment was conducted at the Department of Agronomy,Bangladesh Agricultural University during December 2018 to January 2019.The experiment comprised two factors,(A)Winter rice variety namely,(i)BRRI dhan29 and (ii)BRRI dhan36;(B)Seed priming agent namely (i)Control(no priming),(ii)20000 ppm NaCl,(iii)30000 ppm NaCl,(iv)20000 ppm KCl,(v)30000 ppm KCl,(vi)20000 ppm CaCl2,(vii)30000 ppm CaCl2,(viii)50 ppm CuSO4,(ix)75 ppm CuSO_(4),(x)10000 ppm ZnSO4,(xi)15000 ppm ZnSO4,(xii)2 ppm Na2MoO4,(xiii)3 ppm Na2MoO4,(xiv)100 ppm PEG(Polyethylene glycol 4000)and (xv)150 ppm PEG.Seeds were sown on two different dates viz.,1st December and 1st January so that seedlings are exposed to cold stress at different stages.The experiment was laid out in a completely randomized design(CRD)with three replications.Results indicated that(in most of the cases)seed priming has a positive impact on seedling emergence rate(%),root length,shoot length,root shoot ratio,root dry weight,shoot dry weight,seedling dry weight and survival rate(%).Among the priming agents,KCl and CaCl2 performed best;while priming with NaCl and PEG showed no advantages over no priming for both the sowing dates.In general,BRRI dhan36 performed better than BRRI dhan29 in terms of seedling growth because of its higher tolerance to cold stress.But,both the varieties performed similarly in terms of emergence rate and survival rate.Thus,priming is an effective tool to increase seed germination,better seedling growth,and higher seedling survivability of winter rice under cold stress,and KCl(20000 ppm)or CaCl2(20000 ppm)can be considered as a viable priming agent.

Effects of Silicon on Physiology and Biochemistry of Dendrobium moniliforme Plantlets under Cold Stress

[ Objective] The study aimed to investigate the effects of silicon on physiology and biochemistry of Dendrobium moniliforme plantlets under low tempera- ture stress. [ Method ] By using Dendrobium monilforme as the experimental material, different concentrations of Na2SiO3 （0, O. 2, 0.4, 0.6, 0.8, 1.0, 5.0 and 10.0 mmol/L） was added to the basic medium[ 1/2MS ＋ 6-BA （0.1mg/L） ＋ NAA （ 1 mg/L） ＋ agar （7.2 g/L） ＋ sucrose （30 g/L） ] for tissue culture; af- ter hardening and transplanting, Dendrobium moniliforme plantlets were treated under low temperature stress at 4 ~C for 0, 24 and 48 h, in order to investigate the physiological response of Dendrobium ranniliforme leaves to different concentrations of Na2SiO3. [ Result] Under low temperature stress at 4℃, Dendrob/um mon//i- fortns leaves have certain osmotic regulation ability, and the three osmotic regulation substances show different variation trends at different stages. Appropriate con- centration of NshSiO3 can increase the contents of free proline, soluble sugar and soluble protein to varying degrees, reduce MDA content and further improve the cold resistance of Dendrobium moniliforme plantlets. The order of the effects of Na2SiO~ on various physiological indicators is ： free proline 〉 MDA 〉 soluble sugar （or soluble protein）. According to the correlation analysis among various physiological indicators, free proline, soluble sugar, soluble protein and MDA contents can all be used as reference indicators to identify the cold resistance of Dendrobium moniliforme. [ Conclusion] The addition of Na2SiO3 （0.4 retool/L） can moder- ately decrease the thermal energy for normal growth of Dendrobium moniliforme, which is conducive to reducing the cost of cultivation. Key words Na2SiO3 ;Dendrobium monlifforme;Low temperature stress;Physiological and biochemical characteristics

Proteomic Alterations of Antarctic Ice Microalga Chlamydomonas sp.Under Low-Temperature Stress

Antarctic ice microalga can survive and thrive in cold channels or pores in the Antarctic ice layer. In order to understand the adaptive mechanisms to low temperature, in the present study we compared two-dimensional polyacrylamide gel electrophoresis （2-DE） profiles of normal and low temperature-stressed Antarctic ice microalga Chlamydomonas sp. cells. In addition, new protein spots induced by low temperature were identified with peptide mass fingerprinting based on matrix-assisted laser desorption/ionization time-of-flight mass spectrometry （MALDI-TOF-MS） and database searching. Well-resolved and reproducible 2-DE patterns of both normal and low temperature-stressed cells were acquired. A total of 626 spots was detected in control cells and 652 spots were detected in the corresponding low temperature-stressed cells. A total of 598 spots was matched between normal and stressed cells. Two newly synthesized proteins （a and b） in low temperature-stressed cells were characterized. Protein spot A （53 kDa, pl 6.0） was similar to isopropylmalate/homocitrate/citramalate synthases, which act in the transport and metabolism of amino acids. Protein spot b （25 kDa, pl 8.0） was related to glutathione S-transferase, which functions as a scavenger of active oxygen, free radicals, and noxious metabolites. The present study is valuable for the application of ice microalgae, establishing an ice microalga Chlamydomonas sp. proteome database, and screening molecular biomarkers for further studies.

Contributions of Different Wetland Plants to Nitrogen and Phosphorus Removal in Constructed Wetlands

Constructed wetlands with Cyperus altrnlifolius,Pennisetum sinese Roxb and elephant grass as vegetation were built to study the nitrogen and phosphorous removal from domestic sewage in cold climate.It was found that these three plants could all grow slowly and stably under low temperature stress in winter,and the biomass,nitrogen and phosphorus absorption characteristics and absorbing capacity of the three plants were different.The growth rate and biomass were as follows:P.sinese Roxb>elephant grass>C.altrnlifolius,and these three plants could be used as winter wetland decontamination plants in Southwest China;the absorption characteristics of nitrogen and phosphorus in different tissues of the three plants were as follows:nitrogen content in leaf>nitrogen content in stem,phosphorus content in stem>phosphorus content in leaf;the removal of total nitrogen(TN)by three plants in the wetland was ranged form 4.3%to 7.8%,and the removal of total phosphorus(TP)was ranged from 3.3%to 5.3%.Different plants had different absorption effects on nitrogen and phosphorus.There was a significantly positive correlation between plant nitrogen and phosphorus accumulation and biomass,and nitrogen removal rate in wetland showed a significantly positive correlation with biomass.

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.