cold tolerance

耐寒性
  • 文章类型: Journal Article
    番茄果实(SolanumlycopersicumL.)具有非常短暂的可储存性,在寒冷条件下储存时显示冷害(CI),用于延迟成熟。出于这个原因,在这项研究中,测定了不同浓度(10、50和100mgL-1)的绿原酸(ChA),以评估其在保持番茄果实品质性状和减轻CI症状方面的有效性。我们的结果表明,ChA处理有效地延缓了体重减轻并保持了果实的硬度,在50mgL-1时观察到最佳结果。总的来说,更高的浓度不会导致显著的质量改善.此外,ChA处理的西红柿表现出丙二醛(MDA)含量和电解质泄漏(EL)的降低值,表明改善的膜完整性和减少的氧化损伤。ChA处理在储存过程中也保持了较高的总酚含量(TPC),具有显著水平的单个多酚,例如芦丁,新绿原酸,和对香豆酸,提示抗氧化能力增强,果实品质保存较好。这是首次在任何水果物种中评估ChA降低CI的潜力,它对番茄成熟的影响显示出在冷藏过程中保持水果质量,延长西红柿的储存性。特别是,我们强调其天然来源和有效性作为一种采后治疗。
    Tomato fruit (Solanum lycopersicum L.) has a very brief storability, displaying chilling injury (CI) when stored in cold conditions used to delay ripening. For this reason, in this study, different concentrations (10, 50, and 100 mg L-1) of chlorogenic acid (ChA) were assayed to evaluate its effectiveness in maintaining fruit quality traits and mitigating CI symptoms in tomatoes. Our results showed that ChA treatments effectively delayed weight loss and maintained fruit firmness, with optimal results observed at 50 mg L-1. In general, higher concentrations did not result in significant quality improvements. Additionally, ChA-treated tomatoes exhibited reduced values in malondialdehyde (MDA) content and electrolyte leakage (EL), indicating improved membrane integrity and reduced oxidative damage. ChA treatments also maintained a higher total phenolic content (TPC) during storage, with significant levels of individual polyphenols such as rutin, neochlorogenic acid, and p-coumaric acid, suggesting enhanced antioxidant capacity and better preservation of fruit quality. This is the first time the potential of ChA to reduce CI has been evaluated in any fruit species, and its impact in tomato ripening is shown to uphold fruit quality during cold storage, prolonging the storability of tomatoes. In particular, we highlight its natural origin and effectiveness as a postharvest treatment.
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  • 文章类型: Journal Article
    地中海果蝇(medfly),头皮炎(Wiedemann),来自地中海沿岸栖息地的新鲜水果和蔬菜的最重要的入侵害虫之一,正在将其目前的地理分布扩展到欧洲温度较低的地区。自2010年以来,每年在维也纳地区都会发现这种苍蝇,奥地利。然而,它是否可以建立永久人口尚不清楚。在本文中,C.capitata在维也纳越冬的能力,对奥地利(北纬48.1°)进行了连续两个冬季(2020-2022年)的研究。不同生命阶段的越冬试验(幼虫,蛹,和成年)的C.capitata在露天和没有加热系统的地下室的受保护环境中进行。将对照蝇保持在气候室中的恒定条件下(25°C,60%RH,14:10L:D)。我们的数据表明,地中海果蝇的任何生命阶段都无法在空地上度过奥地利冬季。然而,在受保护的环境中,C.capitata在所有研究的生命阶段中至少在少量的情况下都比冬季更长,并且有几只幸存的雌性能够在接下来的结果季节产卵。讨论了这些发现对欧洲温带国家害虫正在进行的地理范围扩大的影响。
    The Mediterranean fruit fly (medfly), Ceratitis capitata (Wiedemann), one of the most important invasive pests of fresh fruits and vegetables from the coastal Mediterranean habitats, is expanding its current geographic distribution to cooler more temperate areas of Europe. Every year since 2010 the fly is detected in the area of Vienna, Austria. However, whether it can establish permanent populations is not known. In this current paper, the capacity of C. capitata to overwinter in Vienna, Austria (48.1° northern latitude) was studied over 2 consecutive winter seasons (2020-2022). Overwintering trials with different life stages (larva, pupa, and adult) of C. capitata were performed in the open field and in the protected environment of a basement without a heating system. Control flies were kept under constant conditions in a climate chamber (25 °C, 60% RH, 14:10 L:D). Our data showed that no life stage of the Mediterranean fruit fly was able to survive the Austrian winter in the open field. However, in the protected environment C. capitata outlived the winter months in all studied life stages at least in small numbers and several surviving females were able to lay eggs at the time of the following fruiting season. Implications of these findings for the ongoing geographic range expansion of the pest in temperate European countries are discussed.
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  • 文章类型: Journal Article
    对高温和低温变化的适应和耐受性对于植物和动物的生存和增殖至关重要。然而,关于动物和植物之间的共同分子,没有明确的信息。在这项研究中,我们发现热量,线虫秀丽隐杆线虫的耐寒性受RNA结合蛋白EMB-4的相反调节,其植物同源物包含多态性,导致耐热性多样性。秀丽隐杆线虫会根据以前的培养温度改变其对冷和热的耐受性,其中EMB-4通过改变基因表达分别充当耐热和耐冷性的正和负控制器。在表达受EMB-4(一种磷脂加扰酶)调控的基因中,和一种酸性鞘磷脂酶,它们参与膜脂代谢,被发现在耐热性的负调控中起着至关重要的作用。
    Adaptation and tolerance to changes in heat and cold temperature are essential for survival and proliferation in plants and animals. However, there is no clear information regarding the common molecules between animals and plants. In this study, we found that heat, and cold tolerance of the nematode Caenorhabditis elegans is oppositely regulated by the RNA-binding protein EMB-4, whose plant homolog contains polymorphism causing heat tolerance diversity. Caenorhabditis elegans alters its cold and heat tolerance depending on the previous cultivation temperature, wherein EMB-4 respectively acts as a positive and negative controller of heat and cold tolerance by altering gene expression. Among the genes whose expression is regulated by EMB-4, a phospholipid scramblase, and an acid sphingomyelinase, which are involved in membrane lipid metabolism, were found to play essential roles in the negative regulation of heat tolerance.
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  • 文章类型: Journal Article
    由于气候变化,许多地区正在经历越来越温和的冬天。因此,来自温暖地区的害虫,特别是那些对低温有一定耐受性的,可以将其地理范围扩展到这些传统上较冷的地区。棕榈树蛾(Paysandisiaarchon)是一种新热带昆虫,近几十年来,它已成为欧洲和亚洲最严重的棕榈树害虫之一。人们对它能够忍受中度寒冷的冬天的能力知之甚少,因此,殖民新的地区。在这项工作中,我们通过测量其热极限来表征Paysandisiaarchon的耐寒性:中位致死温度,LT50,寒战昏迷发作温度,CTmin,过冷点,SCP,冷冻时间和冷冻存活率。我们发现这个物种能够在短时间内完全冻结,冷冻暴露30分钟后的存活率为87%,和33%的1小时暴露。它是一种中等耐冻的物种,与所有其他产于温暖地区的鳞翅目相反,不耐冻结。此外,我们调查了这种昆虫在短期或长期暴露于亚致死低温后是否提高了其耐寒性。为此,我们研究了主要耐热性参数的潜在变化,用X射线计算机断层扫描,也在预处理动物的形态成分中。我们发现,短暂的暴露前并不意味着SCP和CTmin值的显着变化。相比之下,长期预处理的幼虫提高了它们在冷冻和低温下的存活率,并且需要比其他组更长的时间才能完全冻结。这些长期暴露前的幼虫也出现了一些形态变化,包括可能解释的水含量的减少,至少在某种程度上,它们更长的冷冻时间和更高的冷冻存活率。我们的结果代表了这种害虫的第一个耐寒性特征,这可能与更好的设计策略来对抗它有关。
    Due to climate change, many regions are experiencing progressively milder winters. Consequently, pest insects from warm regions, particularly those with some tolerance to low temperatures, could expand their geographic range into these traditionally colder regions. The palm borer moth (Paysandisia archon) is a Neotropical insect that in recent decades has reached Europe and Asia as one of the worst pests of palm trees. Little is known about its ability to tolerate moderately cold winters and, therefore, to colonize new areas. In this work, we characterized the cold tolerance of Paysandisia archon by measuring its thermal limits: median lethal-temperature, LT50, chill-coma onset temperature, CTmin, supercooling point, SCP, freezing time and freezing survival. We found that this species was able to survive short periods of complete freezing, with survival rates of 87% after a 30-min freezing exposure, and 33% for a 1 h-exposure. It is then a moderately freeze-tolerant species, in contrast to all other lepidopterans native to warm areas, which are freeze-intolerant. Additionally, we investigated whether this insect improved its cold tolerance after either short or long pre-exposure to sub-lethal low temperatures. To that end, we studied potential changes in the main thermo-tolerance parameters and, using X-ray Computed Tomography, also in the morphological components of pretreated animals. We found that short pre-exposures did not imply significant changes in the SCP and CTmin values. In contrast, larvae with long pretreatments improved their survival to both freezing and low temperatures, and required longer times for complete freezing than the other groups. These long-term pre-exposed larvae also presented several morphological changes, including a reduction in water content that probably explained, at least in part, their longer freezing time and higher freezing survival. Our results represent the first cold tolerance characterization of this pest insect, which could be relevant to better design strategies to combat it.
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  • 文章类型: Journal Article
    了解玉米(ZeamaysL.)如何应对寒冷胁迫对于促进耐寒品种的育种计划至关重要。尽管广泛利用全基因组关联研究(GWAS)方法探索与玉米耐寒性相关的有利天然等位基因,很少有报道成功鉴定出有助于玉米耐冷性的候选基因。在这项研究中,通过采用从不同种质来源收集的不同玉米自交系,我们对冷胁迫期间玉米真叶的相对损伤面积的变化进行了GWAS,该性状与玉米耐寒性最密切相关,并鉴定了编码B类热休克转录因子的HSF21,在幼苗和发芽阶段都能积极调节耐寒性。耐寒HSF21Hap1等位基因启动子内的天然变异通过抑制bZIP68转录因子的结合导致冷胁迫下HSF21表达增加,耐寒性的负调节剂。通过整合转录组深度测序,DNA亲和纯化测序,和有针对性的脂质组学分析,我们揭示了HSF21在调节玉米抗寒性的脂质代谢稳态中的功能。此外,HSF21赋予玉米耐寒性而不会导致产量损失。因此,这项研究确立了HSF21作为提高玉米耐寒性的关键调节剂,从而为耐寒玉米品种的选育提供了宝贵的遗传资源。
    Understanding how maize (Zea mays L.) responds to cold stress is crucial for facilitating breeding programs of cold-tolerant varieties. Despite the extensive utilization of the genome-wide association study (GWAS) approach in exploring favorable natural alleles associated with maize cold tolerance, there are few reports that have successfully identified the candidate genes contributing to maize cold tolerance. In this study, by employing a diverse panel of maize inbred lines collected from different germplasm sources, we conducted a GWAS on the variation of the relative injured area of maize true leaves during cold stress-a trait most closely correlated with maize cold tolerance-and identified HSF21, encoding a B-class heat shock transcription factor, which positively regulates cold tolerance at both seedling and germination stages. The natural variations within the promoter of the cold-tolerant HSF21Hap1 allele led to increased HSF21 expression under cold stress by inhibiting the binding of bZIP68 transcription factor, a negative regulator of cold tolerance. Through integrated transcriptome deep sequencing, DNA affinity purification sequencing, and targeted lipidomic analysis, we unveiled the function of HSF21 in regulating lipid metabolism homeostasis for modulating cold tolerance in maize. Additionally, HSF21 confers maize cold tolerance without incurring yield penalties. This study thereby establishes HSF21 as a key regulator that enhances cold tolerance in maize, thus providing valuable genetic resources for the breeding of cold-tolerant maize varieties.
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  • 文章类型: Journal Article
    低温胁迫是影响水稻生长和产量的主要非生物胁迫之一。解剖遗传基础对于水稻耐寒性的遗传改良具有重要意义。在这项研究中,来自云南高原的新的耐寒来源,李江小黑谷,被用作供体亲本,并与冷敏感品种杂交,Deyou17,开发重组自交系(RIL),用于定量性状基因座(QTL)分析,以在水稻的幼苗和孕穗期进行耐寒性。总的来说,确定了2号和7号染色体上幼苗早期的三个耐冷性QTL,以及1号、3号、5号和7号染色体上孕穗期的四个QTL。单倍型和线性回归分析表明,基于这些有利位点的加性效应的QTL金字塔具有良好的耐寒育种潜力。在RIL和BC3F3种群中的效果评估表明,qCTB1在遗传分离种群的孕穗期对耐寒性具有稳定的作用。在不同的冷应激条件下,qCTB1精细映射到标记M3和M4之间的341kb间隔。通过亲本序列的组合比较,基于候选基因的关联分析,以及组织和冷诱导表达分析,确定了qCTB1的8个重要候选基因。本研究将为提高水稻耐寒性的分子育种和基因克隆提供遗传资源。
    在线版本包含补充材料,可在10.1007/s11032-024-01488-3获得。
    Cold stress is one of the main abiotic stresses that affects rice growth and production worldwide. Dissection of the genetic basis is important for genetic improvement of cold tolerance in rice. In this study, a new source of cold-tolerant accession from the Yunnan plateau, Lijiangxiaoheigu, was used as the donor parent and crossed with a cold-sensitive cultivar, Deyou17, to develop recombinant inbred lines (RILs) for quantitative trait locus (QTL) analysis for cold tolerance at the early seedling and booting stages in rice. In total, three QTLs for cold tolerance at the early seedling stage on chromosomes 2 and 7, and four QTLs at the booting stage on chromosomes 1, 3, 5, and 7, were identified. Haplotype and linear regression analyses showed that QTL pyramiding based on the additive effect of these favorable loci has good potential for cold tolerance breeding. Effect assessment in the RIL and BC3F3 populations demonstrated that qCTB1 had a stable effect on cold tolerance at the booting stage in the genetic segregation populations. Under different cold stress conditions, qCTB1 was fine-mapped to a 341-kb interval between markers M3 and M4. Through the combination of parental sequence comparison, candidate gene-based association analysis, and tissue and cold-induced expression analyses, eight important candidate genes for qCTB1 were identified. This study will provide genetic resources for molecular breeding and gene cloning to improve cold tolerance in rice.
    UNASSIGNED: The online version contains supplementary material available at 10.1007/s11032-024-01488-3.
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  • 文章类型: Journal Article
    目标:随着冬季和春季气温的持续升高,在许多季节性寒冷地区,开花和落叶的时机正在推进。这种进步可能会使在春季早期开花的植物在出现晚期冻结事件的年份中面临繁殖减少的风险。不幸的是,对森林群落的花卉抗冻性知之甚少。在这项研究中,我们研究了冻结温度对北美冬季变暖迅速的地区木本植物花朵的影响。
    方法:我们对25种木本植物的花朵进行了硬冷冻(-5ºC)和轻度冷冻(0ºC)。我们使用电解质渗漏评估组织损伤。在物种的一个子集中,我们还研究了硬冻对花粉管生长的影响。为了确定花朵对冰冻伤害的脆弱性是否与开花时间有关,并检查开花时间对春季温度的响应性,我们记录了三年来研究物种的第一朵花的日期。
    结论:跨物种,我们发现花卉的耐冻性与开花时间密切相关,在今年早些时候开花的植物中耐冻性最高。我们假设这些早期开花的物种不太可能受到虚假春天的影响。相反,对假春天最脆弱的物种应该是那些在季节晚些时候开花的物种。这些物种的开花时间对温度也更敏感,让他们冒着经历虚假春天的巨大风险。最终,一年内的花卉损害不会对物种适应性产生大的影响,但是如果假弹簧变得更加频繁,可能会对脆弱物种的繁殖产生长期影响。
    OBJECTIVE: As winter and spring temperatures continue to increase, the timing of flowering and leaf out is advancing in many seasonally cold regions. This advancement could put plants that flower early in the spring at risk of decreased reproduction in years when there are late freeze events. Unfortunately, relatively little is known about floral freezing tolerance in forest communities. In this study, we examined the impact of freezing temperatures on the flowers of woody plants in a region where there is rapid winter warming in North America.
    METHODS: We subjected the flowers of twenty-five woody species to a hard (-5ºC) and a light freeze (0ºC). We assessed tissue damage using electrolyte leakage. In a subset of species, we also examined the impact of a hard freeze on pollen tube growth. To determine if the vulnerability of flowers to freezing damage relates to flowering time and to examine the responsiveness of flowering time to spring temperature, we recorded the date of first flower for our study species for three years.
    CONCLUSIONS: Across species, we found that floral freezing tolerance was strongly tied to flowering time with the highest freezing tolerance occurring in plants that bloomed earlier in the year. We hypothesize that these early blooming species are unlikely to be impacted by a false spring. Instead, the most vulnerable species to a false spring should be those that bloom later in the season. The flowering time in these species is also more sensitive to temperature, putting them at a great risk of experiencing a false spring. Ultimately, floral damage in one year will not have a large impact on species fitness, but if false springs become more frequent, there could be long-term impacts on reproduction of vulnerable species.
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  • 文章类型: Journal Article
    在中国,辽东半岛是蓝莓的重要产区,充足的光,昼夜温差大。然而,辽东半岛冬季和早春期间空气的低温和相对湿度是蓝莓植物受损的主要原因。这里,我们记录了三个蓝莓品种对冷胁迫的转录组和蛋白质组动态响应(“北国”,\'Bluecrop\',和\'Berkeley\')。功能富集分析表明,许多差异表达基因(DEGs)和差异丰度蛋白(DAPs)主要参与内质网蛋白质加工途径。谷胱甘肽代谢途径,和核糖体。我们确定了12,747个转录因子(TFs)分布在20个家族中。根据我们的发现,我们推测,耐寒性的发展是由钙相关基因(CDPKs和CMLs)的表达引起的,谷胱甘肽蛋白质,和TFs(NAC,WRKY,和ERF)。我们的调查发现,三个品种在田间暴露于-9°C至-15°C的温度下会遭受冷害。因此,越冬期间蓝莓的耐寒性不仅要抵抗低温的影响,还要抵抗大风和空气中相对湿度低等复杂环境因素的影响。三个蓝莓品种的抗寒强度顺序为“Berkeley”,\'Bluecrop\',和“北国”。这些结果提供了对冷应激的反应的全面概况,它有可能被用作提高蓝莓耐寒性的项目的选择标记。
    In China, the Liaodong Peninsula is an important growing area for blueberries because of the high organic matter content in the soil, the abundance of light, and the large temperature difference between day and night. However, the low temperature and relative humidity of the air during the winter and early spring in the Liaodong Peninsula are the main reasons for the damage to blueberry plants. Here, we documented the transcriptome and proteome dynamics in response to cold stress in three blueberry cultivars (\'Northland\', \'Bluecrop\', and \'Berkeley\'). Functional enrichment analysis indicated that many differentially expressed genes (DEGs) and differentially abundant proteins (DAPs) were mainly involved in the pathways of protein processing in the endoplasmic reticulum, the glutathione metabolism pathway, and ribosomes. We identified 12,747 transcription factors (TFs) distributed in 20 families. Based on our findings, we speculated that cold tolerance development was caused by the expression of calcium-related genes (CDPKs and CMLs), glutathione proteins, and TFs (NAC, WRKY, and ERF). Our investigation found that three cultivars experienced cold damage when exposed to temperatures between -9 °C and -15 °C in the field. Therefore, the cold resistance of blueberries during overwintering should not only resist the influence of low temperatures but also complex environmental factors such as strong winds and low relative humidity in the air. The order of cold resistance strength in the three blueberry cultivars was \'Berkeley\', \'Bluecrop\', and \'Northland\'. These results provide a comprehensive profile of the response to cold stress, which has the potential to be used as a selection marker for programs to improve cold tolerance in blueberries.
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  • 文章类型: Journal Article
    水稻是全球一半人口的重要食物,对维护全球粮食安全至关重要。气候变化,不断增加的人口和最近的COVID大流行事件造成了财政负担,并威胁着全球粮食安全。由于这些因素,水稻种植也面临着重大挑战。频繁的天气变化对农业规划构成了相当大的挑战,以前是在一致的季节性变化上中继的。在这种情况下,水稻种植对寒冷特别敏感,低温(<18°C)抑制了其发展和生产力。开发温度低,产量好的水稻品种是植物科学家当前育种工作的主要目标之一。为此,短稻和早稻品种最有利于避免寒冷胁迫,并在较少的天数内产量更多。本研究旨在研究低温对不同水稻品种的影响。该研究旨在通过早期和定期培养来鉴定低温粘附基因型。为此,在两个播种季节(2018-2019年)评估了34种基因型,播种时间不同。统计播种时间显示所有产量贡献参数之间的显着相互作用。数据表明,在生殖阶段暴露于低温会延长作物的成熟期,圆锥花序的长度和小穗的生育力也下降了,导致敏感品种产量大幅下降。与其他品种相比,有些品种对冷应激更敏感。在埃及的背景下,Giza176,Sakha104和Sakha107建议用于早期种植,而基因型Giza179,Sakha101,Sakha104和GZ9730-1-1-1-1-1表示为正常培养期。Sakha104品种尤其值得注意,因为这两个目的都推荐。此外,本研究获得的数据为选择适合埃及北三角洲双季种植的水稻品种提供了有价值的信息。这项研究也有助于现有的文献,提供对面对气候变化的水稻种植弹性的见解。
    Rice (Oryza sativa L.) is an essential food for half of the global population and is vital in maintaining global food security. Climate change, increasing population and recent incident of COVID pandemic has generated financial burden and threaten the global food security. Due to theses factors rice cultivation also has to face significant challenges. frequent weather changes pose a considerable challenge to agricultural planning, which was previously relaying on consistent seasonal variations. In this context, rice cultivation is particularly sensitive to cold, where its development and productivity inhibited by low temperatures (< 18 °C). Developing rice varietes with low temprature tolerence and good yield potential is one of the major goals of current breeding efforts of plant scientists. For this purpose, short duration and early rice varieties are most favorable to avoid cold stress and yield more in less number of days. this study was designed to investigate the effect of low temperatures on different rice varieties. the study was designed to identify low temprature tolerent genotypes with early and regular cultivation. For this, thirty-four genotypes were evaluated in two gorwing seasons (2018-2019) with four different sowing times. Statistically sowing time showed significant interaction between all yield contributing parameters. The data indicate that exposure to low temperatures during the reproductive phase prolongs the maturation period of the crop, also length of the panicle and the fertility of the spikelets drops, resulting in a significant decrease in the production of sensitive varieties. Some varieties are more sensitive to cold stress compared to others. In the Egyptian context, Giza176, Sakha104, and Sakha107 are recommended for early cultivation, while the genotypes Giza 179, Sakha101, Sakha104, and GZ 9730-1-1-1-1 are indicated for the normal cultivation period. The Sakha104 variety is particularly notable, as it is recommended for both purposes. In addition, the data obtained in this study provide valuable information for selecting rice varieties suitable for double cropping in the North Delta of Egypt. This study also contributes to the existing literature, providing insights into the resilience of rice cultivation in the face of climate change.
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  • 文章类型: Journal Article
    冷胁迫可以在分子和形态水平上影响植物生物学。我们使用不同的播种方法种植了两种不同类型的烟草幼苗,在4°C下观察到它们的耐寒性的显著差异。冷应激12小时后,浅水播种栽培处理显示出相对较好的生长状态,叶片略有枯萎。使用浮法系统种植的烟草表现得很短,粗根,而那些通过浅水播种种植的人的根细长,有更多的尖端和叉子。在寒冷的压力之后,浅水播种栽培处理表现出更高的抗氧化酶活性,丙二醛(MDA)含量较低。在冷处理的三个阶段(冷胁迫前,在寒冷的压力之后,和恢复3天后)。在分析原始数据后,我们发现,浅水播种栽培处理与低温胁迫前烟酰胺腺嘌呤二核苷酸(NAD)生物合成和NAD代谢的显著功能富集有关,冷应激后与维持细胞结构相关的功能的富集,以及恢复期与光合作用相关的大量功能富集。进行加权基因共表达网络分析(WGCNA),鉴定几个可能导致两种烟草幼苗之间耐寒性差异的hub基因。在我们的分析过程中,与能量转换相关的集线器基因主要在浅水播种栽培处理中得到鉴定,超越其他领域的发现。这些包括AS基因,控制NAD前体的合成,PED1基因,与脂肪酸β-氧化密切相关,和RROP1基因,与ATP生产有关。总的来说,本研究为探索烟苗培育的改良方法提供了有价值的理论依据。通过转录组测序技术,我们已经阐明了在三个时间点不同烟草幼苗中基因表达的差异,确定影响烟草耐寒性的关键基因,为未来的基因编辑提供可能。
    Cold stress can impact plant biology at both the molecular and morphological levels. We cultivated two different types of tobacco seedlings using distinct seeding methods, observing significant differences in their cold tolerance at 4 °C. After 12 h cold stress, shallow water seeding cultivation treatment demonstrates a relatively good growth state with slight wilting of the leaves. Tobacco grown using the float system exhibited short, thick roots, while those cultivated through shallow water seeding had elongated roots with more tips and forks. After cold stress, the shallow water seeding cultivation treatment demonstrated higher antioxidant enzyme activity, and lower malondialdehyde (MDA) content.Transcriptome analysis was performed on the leaves of these tobacco seedlings at three stages of cold treatment (before cold stress, after cold stress, and after 3 days of recovery). Upon analyzing the raw data, we found that the shallow water seeding cultivation treatment was associated with significant functional enrichment of nicotinamide adenine dinucleotide (NAD) biosynthesis and NAD metabolism before cold stress, enrichment of functions related to the maintenance of cellular structure after cold stress, and substantial functional enrichment related to photosynthesis during the recovery period. Weighted gene co-expression network analysis (WGCNA) was conducted, identifying several hub genes that may contribute to the differences in cold tolerance between the two tobacco seedlings. Hub genes related to energy conversion were predominantly identified in shallow water seeding cultivation treatment during our analysis, surpassing findings in other areas. These include the AS gene, which controls the synthesis of NAD precursors, the PED1 gene, closely associated with fatty acid β-oxidation, and the RROP1 gene, related to ATP production.Overall, our study provides a valuable theoretical basis for exploring improved methods of cultivating tobacco seedlings. Through transcriptome sequencing technology, we have elucidated the differences in gene expression in different tobacco seedlings at three time points, identifying key genes affecting cold tolerance in tobacco and providing possibilities for future gene editing.
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