外源海藻糖调节西瓜细胞渗透胁迫抗性的研究

为探究外源海藻糖对渗透胁迫下西瓜细胞生长的影响,本研究以西瓜悬浮培养细胞为试材,测定外源海藻糖预处理后,渗透胁迫对西瓜悬浮培养细胞生长量、细胞外pH、细胞内ROS（活性氧簇）相对含量和微管骨架的影响。结果表明：渗透胁迫下西瓜细胞的生长量明显受到抑制,并且渗透胁迫可诱导西瓜悬浮培养细胞质外体碱化,ROS迸发,细胞微管骨架发生解聚;外源添加海藻糖可在一定程度上缓解渗透胁迫对西瓜悬浮细胞生长量的抑制作用,调节pH、ROS表达水平、并维持微管骨架结构的完整性。以上研究结果表明渗透胁迫下,海藻糖对西瓜细胞具有维持细胞生长、保护亚细胞结构并调节抗性反应的功能。     

An efficient in vitro shoot regeneration through direct organogenesis from seedling-derived petiole and leaf segments and acclimatization of Ficus religiosa

Prolific and rapid in vitro plant organogenesis via direct regeneration has been obtained from axenic seedling-derived petiole and leaf explants of Ficus religiosa in Murashige and Skoog (MS) medium containing different concentrations of cytokinins in combination with indole-3-butyric acid (IBA). MS medium with 1.5 mg/l 6-benzylaminopurine plus 0.15 mg/l IBA produced the highest shoot induction frequency with an average of 6.26 and 10.13 shoots per leaf and petiole explants, respectively. After 4 weeks, the highest root formation frequency (96.7%), root number (5.73), and root length (4.76 cm) were with MS medium containing 2.0 mg/l IBA plus 0.1 mg/l α-naphthalene acetic acid. In addition, the effect of four sodium nitroprusside (SNP) treatments on acclimatization was also studied. Highest morphological traits such as survival rates, fresh and dry root weights as well as antioxidant enzymatic activities such as superoxide dismutase, peroxidase, and catalase was achieved with 125 ppm SNP. The α-amino acid, proline, content was highest with this treatment while the highest H2O2 (hydrogen peroxide) was in the controls. This study introduces a cost-effective, prolific, and efficient in vitro multiplication system to supply pharmaceutical and ornamental needs. It is the first report of an in vitro organogenesis protocol for F. religiosa by direct regeneration through axenic seedling-derived petiole and leaf explants, which can be efficiently employed for the utilization of active biomolecules.     

4种野生早熟禾对土壤酸碱性的生理响应

以甘肃省境内采集的4种野生早熟禾为材料,设8个pH处理(3、4、5、6、7、8、9、10),探讨土壤pH对其生理特性的影响机制以及适宜生长的土壤pH,以期为早熟禾属草坪的养护管理和野生早熟禾的引种驯化提供依据。结果表明,随pH增大,4种野生早熟禾相对膜透性,游离脯氨酸(F-Pro)、可溶性糖(SS)和丙二醛(MDA)含量及过氧化物酶(POD)活性均呈先减小后增大的趋势,可溶性蛋白(SP)含量和超氧化物歧化酶(SOD)、过氧化氢酶(CAT)活性均呈先增大后减小的趋势;草地早熟禾(Poa pratensis)、一年生早熟禾(P.annua)和小药早熟禾(P.micrandra)在pH为7时,硬质早熟禾(P.sphondylodes)在pH为6时受到的伤害最小,是其最适宜生长的pH。     

3种盐胁迫对蓟幼苗生长及生理生化的影响

采用盆栽试验研究不同浓度(0.1%、0.2%、0.3%、0.4%、0.5%、0.6%)的NaCl、Na_2SO_4、Na_2CO_3对蓟(Cirsium japonicum)幼苗苗高、叶片相对含水量、丙二醛(MDA)、抗氧化酶(POD、SOD、CAT)以及渗透调节物质(可溶性糖、游离脯氨酸)的影响。结果表明,除0.1%NaCl处理蓟苗高和叶片相对含水量高于无盐胁迫(CK)外,其余处理均低于CK,且随盐浓度增加而降低,其中,0.3%NaCl、0.4%Na_2SO_4和0.1%Na_2CO_3处理下苗高依次为CK的62.15%、58.00%和45.77%,而0.5%Na_2CO_3处理幼苗死亡;随盐处理浓度增加和处理时间延长,叶片中丙二醛含量均呈上升趋势;POD、SOD、CAT活性随盐浓度、时间变化的趋势不尽一致,3种酶活性均出现了阈值,POD出现在0.3%NaCl、0.3%Na_2SO_4、0.4%Na_2CO_3胁迫5d,SOD出现在0.3%NaCl处理3d、0.5%Na_2SO_4处理1d、0.4%Na_2CO_3处理3d,CAT出现在0.3%NaCl、0.2%Na_2SO_4、0.1%Na_2CO_3胁迫3d;相同处理时间下,随盐浓度的增加,除NaCl胁迫下游离脯氨酸(Pro)含量先增后减外,Na_2SO_4和Na_2CO_3胁迫下游离脯氨酸含量,以及3种盐胁迫下可溶性糖(SS)含量均逐渐增加。综上认为,蓟对NaCl、Na_2SO_4、Na2CO4这3种盐均具有一定的耐性,可以在低于0.3%NaCl、0.4%Na_2SO_4和0.1%Na_2CO_3条件下生存和生长。     

辽棉19号转化耐盐基因AlNHX1的研究

为提高辽宁棉花品种的耐盐性,利用农杆菌介导上胚轴外植体转化法,将来源于盐生植物獐茅的Na~+/H~+逆向转运蛋白基因(AlNHX1)导入棉花辽棉19号中,分析影响棉花上胚轴外植体农杆菌转化的几个重要因素,建立并优化了该品种的转化体系,结果表明:1)农杆菌侵染的最佳时间为90s,共培养过程中乙酰丁香酮最适浓度为100 mg/L,筛选培养基中潮霉素最适浓度为30 mg/L,生根培养基中吲哚乙酸IBA的最适浓度为10mg/L;2)对转化植株进行PCR检测,表明耐盐基因AlNHX1已导入辽棉19号中;3)在盐胁迫下,转基因植株的电导率,渗透势都显著低于未转化植株。结果表明,通过筛选并优化转化体系,辽棉19号品种的耐盐性有较明显的提升,为沿海滩涂地区种植棉花提供了一定的参考。     

绵羊催产素在渗透泵中稳定性的研究

试验旨在评价39.1℃恒温下绵羊催产素在渗透泵（ALZET^®）中的稳定性。试验分为3个组,分别为渗透泵组、微量离心管组和对照组。除对照组始终冻存于-80℃以外,另外两组置于39.1℃培养箱中进行孵育,分不同时间点采集样品。所有样品经超高效液相色谱串联四级杆/飞行时间质谱（ultra-performance liquid chromatography tandem quadrupole/flight time mass spectrometry,UPLC-Q-TOF MS）方法获得氧化产物提取离子流色谱图,通过比较各产物的峰面积,分析催产素在39.1℃缓释泵中的氧化过程。结果表明,催产素在39.1℃缓释泵及微量离心管中均表现出氧化程度随时间逐渐增加的趋势,两试验组之间差异不显著（P＞0.05）。渗透泵组和微量离心管组不同时间点采集样品中催产素原型占对照组原型的百分比分别为83.6%~117.8%和83.2%~116.2%,说明在渗透泵植入绵羊体内试验中需要考虑因催产素氧化引起的含量变化对试验结果的影响。     

胶孢炭疽菌CgSho1基因的克隆与功能分析

HOG-MAPK(high osmolarity glycerol mitogen-activated protein kinase)信号途径是真菌MAPK途径中参与渗透压响应的一条重要通路,在植物病原菌生长发育及致病过程中发挥着重要的作用。Sho1(synthetic high osmolarity-sensitive protein1)是HOG-M APK信号途径上游的一个重要感受器,在不同真菌中常具有不同的功能。本研究从胶孢炭疽菌中克隆了Sho1的同源基因,命名为Cg Sho1,该基因编码一个291个氨基酸的蛋白,含有4个跨膜结构域和一个SH3功能域。利用同源重组的方法获得了该基因的敲除突变体,与野生型相比,敲除突变体表现为营养生长缓慢,菌丝稀疏且疏水性增强,产孢量下降,对氧化压力和渗透压更加敏感,致病力明显减弱。上述结果表明,Cg Sho1参与调控胶胞炭疽菌的营养生长、分生孢子产量、氧化应激反应、渗透压响应及致病性。     

Physiologic response of six plant species grown in two contrasting soils and irrigated with brackish groundwater and RO concentrate

With declining availability of fresh surface water, brackish groundwater is increasingly used for irrigation in the arid and semi-arid southwestern United States. Brackish water can be desalinated by reverse osmosis (RO) but RO results in a highly saline concentrate. Disposal of concentrate is a major problem hindering augmentation of inland desalination in arid areas. The objective of this study was to determine the effect of texture and saline water irrigation on the physiology of six species (Atriplex canescens (Pursh) Nutt., Hordeum vulgare L., Lepidium alyssoides A. Gray, Distichlis stricta (L.) Greene, Panicum virgatum L., and ×Triticosecale Wittm. ex A. Camus [Secale?×?Triticum]). All species were grown in two contrasting soils and irrigated with the same volume of control water (EC 0.9?dS/m), brackish groundwater (4.1?dS/m), RO concentrate (EC 8.0?dS/m). Several plant physiological measurements were made during the growing season including height, number of stem nodes, average internodal length, number of leaves, leaf length, photosynthetic rates, stomatal conductance rates, transpiration rates, leaf temperatures, stem water potential, and osmotic potential. P. virgatum was the only species that showed significant decrease in plant height and growth with texture and irrigation water salinity. Except for A. canescens and L. alyssoides, number and length of leaves decreased with increasing salinity for all species. No significant differences were observed for photosynthetic, stomatal conductance, and transpiration rates by soil texture or irrigation water salinity. Stem water potential and osmotic potential did show some significant influence by soil texture and irrigation water salinity. Based on the results, RO concentrate can be reused to grow all six species in sand; however, growth of all species showed some limitations in clay. Local reuse of RO concentrate along desert margins with regular soil and environmental quality monitoring can accelerate implementation of inland desalination for sustaining food security.     

植物生长调节剂对马铃薯叶片生理代谢及产量品质的影响

大田栽培条件下,以马铃薯克新13为材料,叶面喷施植物生长调节剂2-N,N-二乙氨基乙基己酸酯(Diethyl aminoethyl hexanoate,简称DTA-6)、缩节安(Mepiquat chloride,简称DPC)以及烯效唑(Uniconazole,简称S3307),研究植物生长调节剂对马铃薯叶片生理代谢及产量、品质形成的影响。结果表明:在马铃薯块茎膨大期喷施植物生长调节剂,均显著增加了单薯重、淀粉产量和鲜薯产量,其中,以DTA-6调控效果为最佳,单薯重、淀粉产量、鲜薯产量和淀粉含量比CK分别增加了10.40%、40.26%、26.56%和11.32%,块茎还原糖含量比CK降低了20%。此外,调节剂处理均提高了马铃薯叶片叶绿素含量、净光合速率、蒸腾速率和细胞间隙CO_2浓度,加快了叶片的光合作用,各指标与产量之间呈正相关。随着时间的推移,各处理的叶片淀粉和蔗糖含量呈先降低后升高的变化趋势,淀粉酶活性呈降低的变化趋势,调节剂对叶片蔗糖转化酶活性的调控效果呈不同规律的变化。总体而言,调节剂均提高了马铃薯产量,改善了块茎品质,其中DTA-6处理效果最佳。     

基于超高效液相色谱-高分辨质谱快速筛查绿茶中12种植物生长调节剂的方法研究

建立了超高效液相色谱-高分辨质谱（ultra performance liquid chromatography-high resolution mass spectrometry,UPLC-HRMS）快速筛查绿茶中2,4-滴、对氯苯氧乙酸、吲哚丁酸、氯吡脲和吲哚乙酸等12种植物生长调节剂的方法。采用CAPCELL PKA-C18色谱柱（100 mm×2.1 mm,2 μm）,正离子模式下以体积分数为5%的甲醇水溶液（含有5 mmol/L乙酸铵和体积分数为0.1%甲酸）为流动相;负离子模式下以体积分数为5%甲醇水为流动相,梯度洗脱。在全扫描采集模式下,基于化合物的保留时间、准分子离子峰的精确质量数、碎片离子的精确质量数和同位素分布匹配指数,对目标物进行筛查分析。结果表明:12种植物生长调节剂在绿茶中的报告限（RL）为0.01 mg/kg;在0.01、0.1和0.5 mg/kg 3个添加水平下,其回收率在61%~130%之间,相对标准偏差在1.8%~17%之间。该方法简便快捷,可用于绿茶中常用植物生长调节剂的筛查。