褪黑素引发对干旱胁迫下黄瓜幼苗生理特性的影响

研究褪黑素(MT)种子引发对干旱胁迫下黄瓜幼苗生长的影响,以期为MT作为黄瓜种子引发剂或包衣剂物料筛选提供理论依据。以‘津研4号’为试验材料,用50、100、150 μmol/L MT对种子进行6、12、15 h引发处理,以未引发种子为对照,设置50%田间持水量的干旱胁迫,采用基质盆栽,探究MT引发对干旱胁迫下黄瓜幼苗生长的影响。结果表明,50~100 μmol/L的MT引发15 h时对黄瓜幼苗耐旱性增强有显著作用,其中50 μmol/L时提高幼苗的光合速率达85.5%,超氧化物歧化酶(SOD)、过氧化物酶(POD)、过氧化氢酶(CAT)活性增幅分别为5.1%、42.4%、38.8%,根系活力提高154.0%,说明适宜浓度的MT引发可以通过增强幼苗的抗氧化酶活性及根系活力来提高黄瓜幼苗的耐旱性。     

绵羊毛囊凋亡及抑制的原因初探

为探索在毛囊细胞凋亡中雌激素和褪黑素的作用,实验通过对不同浓度的雌激素和褪黑素水平下体外培养绵羊皮肤的免疫组化染色,发现0.01～0.5 μg/mL雌激素均导致了毛囊和皮脂腺的凋亡,10-4mol/L褪黑素显著抑制了0.01 μg/mL雌激素引起的凋亡,却不能抑制0.5 μg/mL雌激素引起的凋亡.说明雌激素和褪黑素是毛囊周期调控的两个重要激素,它们之间通过相反的作用方式,即通过促进和抑制凋亡来调节毛囊的周期性变化.     

黄瓜采后外源褪黑素处理提高品质和延缓衰老的研究

以采后‘博耐35’黄瓜为试材,研究外源褪黑素浸泡处理对其10℃贮藏期间品质的影响及其机理。结果表明:外源褪黑素处理减缓了黄瓜贮藏过程中叶绿素、维生素C、可滴定酸、可溶性蛋白含量的下降速度,使黄瓜保持了更好的品质。进一步研究得出,外源褪黑素处理降低了黄瓜果实的相对电导率、丙二醛含量、活性氧含量及抗氧化酶(SOD、CAT、POD、APX)活性,降低了对细胞的氧化伤害,并抑制了呼吸速率和乙烯释放量,使细胞保持了更完整的结构,抑制了线粒体和叶绿体的形变。100μmol·L~(-1)的外源褪黑素处理可使黄瓜保持更好的品质,最大程度降低对细胞的氧化伤害,延缓了衰老,达到延长货架期的目的。     

外源褪黑素对盐胁迫下棉花幼苗生长及光合特性的影响

【目的】土壤盐渍化严重制约棉花幼苗生长,褪黑素(Melatonin,MT)能有效调控植物在盐胁迫下的生长机制。研究外源褪黑素对盐胁迫下棉花幼苗生长及光合特性的影响。【方法】采用对棉花幼苗叶面喷施外源褪黑素的方法,分析外源褪黑素在NaCl胁迫下棉花幼苗生长及光合系统的响应,筛选出最适喷施褪黑素浓度。以新陆中70号为材料,设置不同褪黑素浓度(25、50、100、200 μmol/L)处理,测定NaCl胁迫下不同时间段(1、3、6和9 d)棉花幼苗生长指标及叶片荧光、光合作用参数等指标。【结果】NaCl胁迫显著降低棉花幼苗株高、叶片光合、荧光指标,喷施外源褪黑素后,促进株高生长及干物质质量的增加,并有效缓解了NaCl胁迫对棉花幼苗叶片净光合速率、气孔导度、最大光化学效率、PSⅡ有效光量子产量、光化学猝灭等指标的影响,减少NaCl胁迫对棉花幼苗的损伤。【结论】NaCl胁迫条件下,外源褪黑素能有效促进棉花幼苗生长,喷施100 μmol/L褪黑素浓度处理效果最佳。     

采后褪黑素处理对鲜切花椰菜保鲜品质及货架期的影响

为研究采后外源性褪黑素处理对鲜切花椰菜货架期间品质及生理的影响，试验以"雪白"花椰菜为材料，拟从中筛选出有效的使用浓度，进而从生理、细胞和基因表达水平解析褪黑素对鲜切花椰菜保鲜品质的调节作用，以期为探索鲜切花椰菜保鲜和衰老调控的有效途径提供科学依据。采用0.05、0.10、0.50 mmol/L褪黑素溶液浸泡鲜切花椰菜样本15 min，分析货架期间（0、4、8、12、16 d）样本的失重率、硬度、1,1-二苯基-2-三硝基苯肼（1,1-diphenyl-2-picrylhydrazyl, DPPH）值、总酚含量、抗坏血酸含量、色泽、总硫代葡萄糖苷含量、内源性褪黑素含量、多聚半乳糖醛酸酶（polygalacturonase, PG）和脂氧合酶（lipoxygenases, LOX）活性及基因表达水平、细胞超微结构的变化。结果表明：0.10 mmol/L褪黑素处理明显延缓（P<0.05）了失重率、硬度、色泽和总硫代葡萄糖苷含量的下降，提升（P<0.05）了样本中内源性褪黑素的含量。同时，0.10 mmol/L褪黑素处理最有利于提高总酚含量和抗氧化能力，延缓抗坏血酸的降低。此外，0.10 mmol/L可抑制细胞结构的损伤，降低PG和LOX的基因表达，延缓PG和LOX的活性。由此可知，对鲜切花椰菜进行外源褪黑素处理可能是延长其采后寿命和提高品质的有效技术。     

Effect of chelating and antioxidant agents on morphology and DNA methylation in freeze-drying rabbit (Oryctolagus cuniculus) spermatozoa

Freeze-drying (FD) has been exhaustively tried in several mammalian species as an alternative technique to sperm cryopreservation, but few studies have been done in rabbits (Oryctolagus cuniculus). The main objective of this study was to compare the protective effect of various antioxidants added to EDTA medium on structural and functional components of FD rabbit spermatozoa and on their status of global DNA methylation. FD media used were composed of basic FD medium (10 mM Tris-HCl buffer and 50 mM NaCl) supplemented with either 50 mM EDTA alone (EDTA) or added with 105 µM of rosmarinic acid (RA, EDTA-RA) or 10 µM of melatonin (MLT, EDTA-MLT). The effect of each medium on the preservation of FD spermatozoon structure was evaluated with light and scanning electron microscopy (SEM). Global DNA methylation was quantified in all FD sperm samples as well as in fresh spermatozoa. Morphologically, fracture points were evidenced in the neck, mid and principal piece of the spermatozoon tail. No differences in spermatozoon fracture points were evidenced among FD treatments: intact spermatozoa were the largest (p < .01) category, whereas the most frequent (p < .01) injury was the neck fracture, resulting in tailless heads. At SEM, the head of spermatozoa showed a well-conserved shape and intact membrane in all treatments. DNA methylation status was the same in all FD treatments. In conclusion, supplementation of EDTA, EDTA-RA and EDTA-MLT during FD preserved rabbit sperm morphological integrity and methylation status as well. Therefore, the difficulty of getting viable offspring using FD semen is likely unrelated to the impact of the lyophilization process on DNA methylation and morphology of lyophilized spermatozoa.     

Age-related changes in 2-[125I]-iodomelatonin binding sites in the brain of sea breams (Sparus aurata,L.)

The pineal organ of fish, through its 24h rhythmic release of melatonin, acts as a transducer of the photoperiod, influencing different physiological functions (e.g., reproduction, growth). The target sites for melatonin are poorly known in fish, especially marine species. A radioligand study was undertaken using the gilthead sea bream (Sparus aurata) maintained under natural temperature and photoperiod (at 28°N latitude). This species exhibits the property of changing sex during growth. Brains of one year-old males were collected at 16:00h and brains of three year-old females at 03:00, 10:00, 16:00 and 23:00h. Membrane homogenate receptor assays were run using 2-[¹²⁵I]iodomelatonin as a ligand. Binding sites were detected in brains of young and old fish. In the younger, the exhibited a B_max between 3.52 and 4.29 fmol mg protein^–1 and a K_D between 358–380 pmol l^–1. In the older fish, the K_D varied according to a daily pattern: values were three times higher at 03:00 and 10:00h (500–600 pmol l^–1) than at 16:00 and 23:00h (150–300 pmol l^–1). The number of sites also were higher at 03:00 and 10:00h (180–200 fmol mg protein^–1) than at 16:00 and 23:00h (95–110 fmol mg protein^–1). Melatonin and iodomelatonin displaced 2-[¹²⁵I]iodomelatonin binding in a dose dependent manner, the second being more potent than the first. Binding was also inhibited by GTP. The results provide the first evidence for the presence of membrane melatonin binding sites in the brain of an exclusively marine fish. They suggest that their number and affinity varies during growth and throughout a light/dark cycle. Future experiments will aim to precise the anatomical location and role of these binding sites.     

Melatonin alleviates cold-induced oxidative damage by regulation of ascorbate–glutathione and proline metabolism in melon seedlings (Cucumis melo L.)

Cold stress is one of the most detrimental environmental factors affecting plant growth and development. Melatonin (MEL), a natural indoleamine compound, responds to various environmental cues. To explore the role of MEL in the response of melon (Cucumis melo L.) seedlings to cold stress, the effects of exogenous MEL on the ascorbate–glutathione (AsA–GSH) cycle and proline metabolism were investigated. Melon seedlings were sprayed with various concentrations of MEL (0, 50, 100, 200, or 400 μM), then exposed to cold stress, 12/6°C (day/night) for 7 d, followed by recovery at 28/18°C for another 7 d. The results showed that MEL, especially the 200 μM treatment, dramatically alleviated growth inhibition caused by cold stress, manifested by increased plant growth and decreased O₂^?¯ production rate and malondialdehyde content. Importantly, exogenous application of MEL enhanced the ratios of reduced and oxidized forms of AsA (AsA/DHA) and GSH (GSH/GSSG), and the activities of ascorbate peroxidase (APX), glutathione reductase (GR), and dehydroascorbate reductase (DHAR) involved in the ascorbate–glutathione (AsA–GSH) cycle in melon seedings grown under cold stress. Besides, MEL pretreatment further increased the contents of proline and soluble protein under cold stress. The results reveal that protective effects of MEL against cold stress in melon seedlings are most likely associated with the regulation of the AsA–GSH cycle and proline metabolism as an effective antioxidant system.     

Supplementation of corn seed with regular diet modulates immune function and antioxidant status in Capra hircus

Melatonin modulates the functional activation of antioxidative enzymes to maintain the oxidant–antioxidant balance in the physiological system. Melatonin administration via subcutaneous injection increases endogenous melatonin concentration that has been used to modulate the immune function in rodents and reproductive activity in small ruminant like goats. However, to date, no report exist which could suggest the effect of corn seed supplementation on the endogenous melatonin concentration and its impact on cellular‐immune function and antioxidative enzymes activity in any small ruminant like goats. Therefore, in this study, Zea mays (corn seeds), as source of melatonin, were supplemented with regular diet to explore its impact on endogenous melatonin concentration in goat, Capra hircus. Zea mays supplementation with regular diet for 40 days significantly elevated the endogenous melatonin concentration, proliferative response of peripheral blood mononuclear cells and antioxidative enzymes activity along with total antioxidant capacity of the plasma. An increase in circulatory IL‐2 and IL‐6 level along with declined TNF‐α, malondialdehyde and nitric oxide was noted with elevated endogenous melatonin concentration. In conclusion, it might be suggested that corn seed could be used as supplement to modulate endogenous melatonin concentration that may improve cellular‐immune function and antioxidant status. The corn seed might also be included as nutraceutical in regular diet to maintain immune‐antioxidant homoeostasis in other species.