转AtNHX1基因玉米的产生及其耐盐性分析

以玉米(ZeamaysL.)骨干自交系DH4866、齐319和鲁原16106的胚性愈伤组织为材料,采用农杆菌介导法将AtNHX1和hpt基因转入玉米培养细胞,经筛选获得了抗潮霉素的愈伤组织并再生植株。经PCR检测和Southernblot验证,确定了22.8%的再生植株为转基因植株。农杆菌液浓度、愈伤组织基因型及共培养时间对转化率均有明显影响。外源基因在转基因植株后代中的分离呈多样性,在部分株系中表现出孟德尔遗传规律。耐盐筛选表明,一些转基因植株及其后代具有很好的耐盐性,部分株系可在0.8%-1.0%NaCl溶液浇灌下萌发和生长。Northern杂交表明,植株耐盐性提高与AtNHX1基因的转录水平相一致。     

Salt accumulation in vegetative organs and ecological stoichiometry characteristics in typical halophytes in Xinjiang,China

AimsStudying salt accumulation in vegetative organs and carbon (C), nitrogen (N), phosphorus (P) stoichiometry in halophytes contributes to understand the adaptive strategy to saline environment and the distribution mechanism of nutrients and salinity of the halophytes.MethodsFour typical dominant species, including three perennial shrubs (Halocnemum strobilaceum, Suaeda microphylla and Kalidium foliatum) and an annual herb (Salicornia europaea) were selected for analyzing the salt accumulation and C, N, P stoichiometry relationships.Important findings 1) The salt accumulation showed “salt island” effect at peak growing stage; Na⁺, Cl^- and electrical conductivity all showed significant positive trends, when salt ions were transported from the root to the shoot of 4 halophytes. 2) P was the limiting growth element of perennial shrubs, while both N and P were limiting factors for the annual herb. 3) Different organs, species and their interactions affected C, N, P stoichiometry and salt icons except Mg²⁺. 4) C was negatively correlated with N and P, but there was a significant positive correlation between N and P. 5) Electrical conductivity, Na⁺and K⁺ were positively correlated with N, N:P ratio, and negatively correlated with C, C:N and C:P ratios, and yet K⁺, CO₃^2- had significant positive correlation with P. These results implied that there were some kind of mutual promoting relationships between nutrients and salinity in halophytes in the saline environment, and fertilizing with N could promote sodium chloride desalination.     

Multi-target novel neuroprotective compound ITH33/IQM9.21 inhibits calcium entry, calcium signals and exocytosis

Compound ITH33/IQM9.21 (ITH/IQM) belongs to a new family of l-glutamic acid derivatives with antioxidant and neuroprotective properties on in vitro and in vivo models of stroke. Because neuronal damage after brain ischemia is tightly linked to excess Ca²⁺ entry and neuronal Ca²⁺ overload, we have investigated whether compound ITH/IQM antagonises the elevations of the cytosolic Ca²⁺ concentrations ([Ca²⁺]_c) and the ensuing exocytotic responses triggered by depolarisation of bovine chromaffin cells. In fluo-4-loaded cell populations, ITH/IQM reduced the K⁺-evoked [Ca²⁺]_c transients with an IC₅₀ of 5.31 μM. At 10 μM, the compound decreased the amplitude and area of the Ca²⁺ transient elicited by challenging single fura-2-loaded cells with high K⁺, by 40% and 80%, respectively. This concentration also caused a blockade of K⁺-induced catecholamine release at the single-cell level (78%) and cell populations (55%). These effects are likely due to blockade of the whole-cell inward Ca²⁺ currents (IC₅₀ = 6.52 μM). At 10 μM, ITH/IQM also inhibited the Ca²⁺-dependent outward K⁺ current, leaving untouched the voltage-dependent component of I_K. The inward Na⁺ current was unaffected. Inhibition of depolarisation-elicited Ca²⁺ entry, [Ca²⁺]_c elevation and exocytosis could contribute to the neuroprotective effects of ITH/IQM in vulnerable neurons undergoing depolarisation during brain ischemia.     

外源cGMP调控盐胁迫下黑麦草种子萌发机制

c GMP(cyclic guanosine-3',5'-monophosphate)是一类环化核苷酸,能响应植物非生物胁迫的信号分子。以c GMP的膜透性类似物8-Br-cGMP为供体,研究c GMP对盐胁迫下黑麦草种子萌发及生理指标的影响。通过计算萌发指标、测定种子萌发过程中各项生理指标,探讨c GMP对黑麦草种子萌发时耐盐机制的作用。结果表明:低浓度(50 mmol/L)盐胁迫能促进黑麦草种子的萌发,随着盐浓度(100、150、200、250 mmol/L)的增加黑麦草种子的萌发率逐渐降低甚至抑制其萌发。而加入20μmol/L c GMP能缓解100 mmol/L Na Cl胁迫对黑麦草种子造成的伤害,使黑麦草种子的发芽率、发芽势、发芽指数、活力指数分别提高了7.4%、133.3%、52.1%、104.2%。此外,研究发现加入20μmol/Lc GMP能够促进100 mmol/L Na Cl盐胁迫下黑麦草萌发期根的生长,比单独100 mmol/L Na Cl盐胁迫处理下的黑麦草植株全长、根长、叶长分别提高了2.5、2.8倍和2.6倍,种子在萌发过程中可溶性糖、可溶性蛋白含量、淀粉酶活性,脯氨酸含量分别提高了47.9%、15.7%、94.3%、117.4%,而淀粉含量、MDA含量、电导率,O_2~-产生速率分别降低了40.9%、128.7%、88.6%、211.9%。说明20μmol/Lc GMP通过提高可溶性糖,可溶性蛋白含量及脯氨酸含量,同时减少MDA、O_2~-的产生,以此缓解盐胁迫对黑麦草种子的伤害、同时促进淀粉水解,从而加速种子萌发。     

Differential effects of four abiotic factors on the germination of salt marsh annuals

Interspecific differences in responsiveness to temperature, photoperiod, soil salinity, and soil moisture confirm the hypothesis that abiotic factors differentially affect the germination of salt marsh plants. In growth chamber experiments, four of eight annual species responded to small differences in temperature or photoperiod. Increasing soil salinity decreased the final proportion of seeds germinating and slowed germination for each of the seven species tested. Higher soil moisture increased the proportion germinating of five species and germination speed of all seven species. Salinity and moisture interacted to affect the proportion germinating of five species and germination speed of all seven species. Although the abiotic factor with the largest effect on germination varied among species, more species responded to, and the magnitudes of the responses were larger for, soil salinity than for the other abiotic factors. These germination tests partially explained interspecific differences in the timing of germination in the field. Patterns of Hutchinsia procumbens, Lythrum hyssopifolium, Parapholis incurva, and possibly Lasthenia glabrata ssp. coulteri germination in response to a nonseasonal rainfall could be explained by their response to salinity, temperature, or photoperiod. Fine-scale differences in the timing of establishment within the typical germination window and spatial distributions along salinity and moisture gradients were not explained.     

内生细菌252和254对盐胁迫下小麦幼苗过氧化物酶和过氧化氢酶活性的影响

以‘周麦18’为试验材料,采用盆栽法设置盐胁迫、盐胁迫下接菌和对照3组处理,研究不同盐浓度胁迫下接种内生菌对小麦幼苗生长的影响.结果表明: 内生菌252、254分别与死亡谷芽孢杆菌和根癌土壤杆菌具有最高相似率.盐胁迫下接种内生细菌252,在14 d时小麦幼苗随盐浓度升高过氧化物酶(POD)活性先升高后下降,其中盐浓度为100 mmol·L^-1时POD活性最高(13370 U·g^-1·min^-1);21 d时小麦幼苗POD和过氧化氢酶(CAT)活性较对照明显升高;28 d时盐浓度为150、250 mmol·L^-1的POD和CAT活性最高.盐胁迫下接种内生细菌254,在14 d时盐浓度为250 mmol·L^-1的小麦幼苗CAT活性最高;21 d时盐浓度在100～250 mmol·L^-1的POD和CAT活性较高,其中150 mmol·L^-1时CAT活性最高;28 d时盐浓度为200 mmol·L^-1的CAT活性最高.在盐胁迫下,接种内生菌对小麦幼苗不同培养时期发挥最佳作用的盐浓度不同.内生菌252和254可提高盐胁迫下小麦植株POD、CAT活性,对小麦幼苗生长有一定修复作用.     

Mycosporines and mycosporine-like amino acids: UV protectants or multipurpose secondary metabolites?

Mycosporines and mycosporine-like amino acids (MAAs) are low-molecular-weight water-soluble molecules absorbing UV radiation in the wavelength range 310-365 nm. They are accumulated by a wide range of microorganisms, prokaryotic (cyanobacteria) as well as eukaryotic (microalgae, yeasts, and fungi), and a variety of marine macroalgae, corals, and other marine life forms. The role that MAAs play as sunscreen compounds to protect against damage by harmful levels of UV radiation is well established. However, evidence is accumulating that MAAs may have additional functions: they may serve as antioxidant molecules scavenging toxic oxygen radicals, they can be accumulated as compatible solutes following salt stress, their formation is induced by desiccation or by thermal stress in certain organisms, they have been suggested to function as an accessory light-harvesting pigment in photosynthesis or as an intracellular nitrogen reservoir, and they are involved in fungal reproduction. Here, the evidence for these additional roles of MAAs as 'multipurpose' secondary metabolites is reviewed, with special emphasis on their functions in the microbial world.     

Xanthine oxidase converts nitric oxide to nitroxyl that inactivates the enzyme

Xanthine oxidase (XO) was found to convert nitric oxide (NO* ) released from spermine-NONOate to nitroxyl (HNO), the one-electron reduction product of NO*, in the presence of its substrate hypoxanthine under anaerobic conditions. Under these conditions, XO lost its activity. Upon aerobic incubation of XO with its substrate, neither conversion of NO* to HNO nor inactivation of the enzyme was observed. Angeli's salt (an HNO generator) or synthetic peroxynitrite inactivated XO at low concentrations, whereas high concentrations of diethylamine-NONOate (an NO* donor) and SIN-1 (which generates peroxynitrite by releasing both NO* and superoxide) were required to inactivate XO. These results suggest that HNO generated by XO under anaerobic conditions inactivates XO. As both XO and NO* synthase are activated and/or induced in ischemia-reperfusion injury, HNO formed by XO may contribute to pathogenesis by exerting its potent oxidation activity against a variety of biological compounds.     

生长素和氯化钙对盐胁迫下沙冬青幼苗的缓解作用

沙冬青(Ammopiptanthus mongolicus)是亚洲中部荒漠地区唯一的常绿阔叶灌木,本课题组已将其在天津引种栽培成功。为进一步了解生长素和氯化钙对沙冬青在盐胁迫下的缓解作用,用水培方法培养沙冬青幼苗,测定其在1.3%NaCl胁迫下经不同浓度的生长素和氯化钙处理后抗氧化酶活性、PSII光化学效率以及其它与植物抗性有关的生理指标的变化。结果表明,一定浓度的IBA和CaCl2处理可以促进盐胁迫下沙冬青幼苗的生长、提高叶绿素的含量、增强保护酶的活性、降低丙二醛含量,缓解了盐胁迫。但是当IBA、CaCl2浓度分别达到5和20mmol.L-1时则对沙冬青幼苗产生伤害。综合各项生理指标说明,CaCl对沙冬青盐胁迫的缓减作用要优于IBA。