胡杨细胞和组织结构与其耐盐性关系的研究

利用电子显微镜和光学显微镜中相差和微分干涉等技术对胡杨（ＰｏｐｕｌｕｓｅｕｐｈｒａｔｉｃａＯｌｉｖ．）悬浮培养的细胞和试管苗的组织和细胞结构以及盐胁迫对其结构影响进行了研究。与毛白杨（Ｐ．ｔｏｍｅｎｔｏｓａＣａｒ．）相比,胡杨存在与耐盐相关的结构,如根尖具有较发达的表皮和外皮层,根毛着生靠近根端;叶片输导组织不发达;细胞中线粒体和质体较丰富。盐胁迫和渗透胁迫可使胡杨细胞质中的线粒体和质体变得更为丰富,质体中内含体增多,在细胞质中和液泡内缘出现明显的嗜锇物质,在悬浮培养的细胞内表现出明显的丝状结构,细胞核变大,核仁明显。受到８ｇ／ＬＮａＣｌ胁迫的胡杨根尖分生细胞仍可维持正常的结构。在超微结构研究中还发现,胡杨细胞膜与细胞壁之间呈齿状结合,说明了膜与壁之间结合的牢固性和稳定性,解释了胡杨细胞在胁迫中不易发生质壁分离的原因。     

Osmotic Stress and Ion-Specific Effects on Xylem Abscisic Acid and the Relevance to Salinity Tolerance in Poplar

We designed two experiments to investigate the osmotic stress and ion-specific effects on xylem abscisic acid (ABA) and the relevance to salinity tolerance in one-year-old seedlings of Populus euphratica Oliv. (a salt-resistant genotype) and one-year-old rooted cuttings of P. 'popularis 35-44' (P. popularis) (a salt-sensitive genotype). Net photosynthetic rates (Pn) and unit transpiration rates (TRN) of the two genotypes were significantly decreased upon osmotic shock caused by PEG 6000 (osmotic potential = -0.24 MPa) or iso-NaCl (50 mM). Shoot xylem ABA concentrations in both genotypes increased rapidly after the onset of PEG stress, resulting from a decreased water flow. NaCl-treated trees of P. euphratica maintained considerably greater concentrations of ABA than PEG-treated plants in a longer term, whereas salinized P. popularis exhibited a transient accumulation of ABA in the shoot. TRN was greatly enhanced in both genotypes when pressure (0.24 MPa) was applied to counteract the osmotic suction of 50 mM NaCl. Pressurizing of root systems diluted solutes in the root xylem, but the dilution effect was more pronounced in P. popularis. Root xylem ABA concentrations in P. euphratica steadily increased with salt stress although pressurization lowered its levels. In contrast, there were no observed changes in ABA response to salinity in pressured P. popularis. Therefore, we concluded that the salt-tolerant P. euphratica had a greater capacity to synthesize ABA under saline conditions, which may partially result from specific salt effects. In addition, P. euphratica exhibited a higher capacity for salt (Na+ and Cl-) transport control under salt stress, compared with P. popularis. The possible association between ABA and salt transport limitation, and the relevance to salinity tolerance were discussed.     

Osmotic Stress and Ion-Specific Effects on Xylem Abscisic Acid and the Relevance to Salinity Tolerance in Poplar

We designed two experiments to investigate the osmotic stress and ion-specific effects on xylem abscisic acid (ABA) and the relevance to salinity tolerance in one-year-old seedlings of Populus euphratica Oliv. (a salt-resistant genotype) and one-year-old rooted cuttings of P. 'popularis 35-44' (P. popularis) (a salt-sensitive genotype). Net photosynthetic rates (Pn) and unit transpiration rates (TRN) of the two genotypes were significantly decreased upon osmotic shock caused by PEG 6000 (osmotic potential = -0.24 MPa) or iso-NaCl (50 mM). Shoot xylem ABA concentrations in both genotypes increased rapidly after the onset of PEG stress, resulting from a decreased water flow. NaCl-treated trees of P. euphratica maintained considerably greater concentrations of ABA than PEG-treated plants in a longer term, whereas salinized P. popularis exhibited a transient accumulation of ABA in the shoot. TRN was greatly enhanced in both genotypes when pressure (0.24 MPa) was applied to counteract the osmotic suction of 50 mM NaCl. Pressurizing of root systems diluted solutes in the root xylem, but the dilution effect was more pronounced in P. popularis. Root xylem ABA concentrations in P. euphratica steadily increased with salt stress although pressurization lowered its levels. In contrast, there were no observed changes in ABA response to salinity in pressured P. popularis. Therefore, we concluded that the salt-tolerant P. euphratica had a greater capacity to synthesize ABA under saline conditions, which may partially result from specific salt effects. In addition, P. euphratica exhibited a higher capacity for salt (Na+ and Cl-) transport control under salt stress, compared with P. popularis. The possible association between ABA and salt transport limitation, and the relevance to salinity tolerance were discussed.     

胡杨木质部水分传导对盐胁迫的响应与适应北大核心CSCD

解析植物木质部导水率对逆境的响应和适应对促进植物抗逆性机理研究和受损植被恢复具有重要意义。该文以荒漠河岸林建群种胡杨(Populus euphratica)为研究对象,系统分析了胡杨幼株根、茎、叶水分传输通道对不同浓度盐胁迫的响应和适应。结果表明:(1)胡杨幼株根系对盐胁迫的敏感性高于茎和叶,盐胁迫下根系生长和根尖数显著受到抑制,根木质部易于发生栓塞,导水率明显降低。(2)胡杨幼株茎木质部导水率对盐胁迫的响应依盐浓度而定,轻度(0.05 mol·L–1 Na Cl)和中度(0.15 mol·L–1 Na Cl)盐胁迫下,胡杨可以通过协调导管输水的有效性和安全性来调节木质部的导水率,维持植物正常生长;重度(0.30 mol·L–1 Na Cl)盐胁迫下,胡杨茎木质部导管输水有效性和安全性均明显降低,木质部导水率显著下降,并伴随叶片气孔导度的显著降低,从而严重抑制了胡杨的光合和生长。     

Adaptation to high salinity in poplar involves changes in xylem anatomy and auxin physiology

To investigate the physiological basis of salt adaptation in poplar, we compared the effect of salt stress on wood anatomy and auxin physiology of the salt-resistant Populus euphratica and salt-sensitive Populus x canescens. Both poplar species showed decreases in vessel lumina associated with increases in wall strength in response to salt, however, in P. euphratica at three-fold higher salt concentrations than in P. x canescens. The predicted hydraulic conductivity of the wood formed under salt stress decreased in P. x canescens, while in P. euphratica, no significant effects of salt on conductivity and transpiration were observed. The concentration of free indole-3-acetic acid (IAA) decreased under salt stress in the xylem of both poplar species, but to a larger extent in P. x canescens than in P. euphratica. Only salt-treated P. euphratica exhibited an increase in IAA-conjugates in the xylem. Genes homologous to the auxin-amidohydrolase ILL3 were isolated from the xylems of P. euphratica and P. x canescens. For functional analysis, the auxin-amidohydrolase from P. x canescens was overexpressed in Arabidopsis. Transgenic Arabidopsis plants were more resistant to salt stress than the wild-type plants. Increased sensitivity of the transgenic Arabidopsis to IAA-Leu showed that the encoded hydrolase used IAA-Leu as a substrate. These results suggest that poplar can use IAA-amidoconjugates in the stem as a source of auxin to balance the effects of salt stress on auxin physiology.     

胡杨无性系幼苗响应盐胁迫的miRNA表达差异研究

植物miRNA在调控基因表达、细胞周期、生物体发育、抗逆等方面起重要作用。为研究胡杨(Populus euphratica Oliv.)的耐盐机制,以1年生胡杨无性系幼苗为材料,构建具有空间代表性的盐胁迫胡杨cDNA文库,利用二代测序技术测定NaCl胁迫下和正常培养条件下胡杨叶和根miRNA表达情况。结果表明,不同的miRNA之间表达量存在明显差异,表达丰度最高的miRNA有miR156、miR157、miR165、miR166和miR167等,合计占总表达量的90%以上。胡杨根部存在特异表达的miRNA,在整个耐盐调控机制中发挥着生理调节、分子调控和信号传导等极为重要的作用。盐处理样品中发现大量响应盐胁迫的miRNA,对这些转录因子进行靶基因预测和注释后,发现很多盐胁迫响应的miRNA与NAC和SPL等重要转录因子家族相关,与前人的结论一致,另外还发现许多miRNA的调控对象是ATP酶和激素响应因子。     

多效唑浸种对渗透胁迫下玉米根边缘细胞发生的影响

以‘农大108’玉米种子为材料,采用不同浓度(50、100、150、200、250、300、350 mg·L-1)的多效唑溶液进行浸种处理,研究其对渗透胁迫(20％ PEG-6000)下玉米种子萌发、根系生长和根系边缘细胞数目、活性及黏胶层厚度的影响.结果表明:与对照相比,渗透处理抑制了玉米种子露白与根系生长,增加了边缘细胞数目与黏胶层厚度.预先用多效唑浸种后再进行渗透胁迫处理进一步降低了主根的长度,但增加侧根的生长而使根系鲜重增加,进一步增加了根边缘细胞黏胶层厚度,在一定程度上减少了由于渗透胁迫造成的边缘细胞数目的增加程度.无论是渗透处理还是预先用多效唑浸种处理对边缘细胞活性的影响均不大.可见,多效唑浸种能够增加玉米根系的抗旱能力与边缘细胞黏胶层厚度的增加有关,而与边缘细胞数目、活性的关系不大;多效唑浸种溶液的适宜浓度范围为200～250 mg·L-1.     

Lanthanum Prevents Salt Stress-induced Programmed Cell Death in Rice Root Tip Cells by Controlling Early Induction Events

In a previous study, a salt stress-induced programmed cell death （PCD） model was established in rice root tip cells. Here, by using Wuyunjing 8th rice seedlings, the effects of lanthanum on salt stress-induced PCD early events were studied. The results indicated that low concentrations （10 μmol/L）, but not high concentrations （100 μmol/L） of LaCl3 could effectively prevent salt stress-induced PCD. Further study demonstrated that in the early stages of salt-induced PCD process, 10 μmol/L of La^3＋ could prevent the increase of cytoplasmic calcium levels, inhibit reactive oxygen species （ROS） production, and enhance the ROS-scavenging enzyme activities such as superoxide dismutases （SOD） and ascorbate peroxidase （APX）. Imidazole （20 mmol/L）, the inhibitor of nicotinamide adenine dinucleotide phosphate-oxidase （NADPH oxidase）, could alleviate the occurrence of PCD obviously, and such alleviation could be enhanced by the addition of La^3＋, indicating the involvement of NADPH oxidase in the salt stress-induced PCD process. Taken together, lanthanum could prevent salt stress-induced PCD occurrence in the rice root tip cells by blocking the calcium influx under stress, which was followed by inhibiting calcium-dependent NADPH oxidase activity to prevent O2^·- production and, enhancing the cytosolic antioxidative enzyme activities to scavenge the reactive oxygen species.     

Salt Stress-induced Programmed Cell Death in Rice Root Tip Cells

Salt stressed rice root tips were used to investigate the changes of reactive oxygen species （ROS） and antioxidant enzymes at the early stages of programmed cell death （PCD）. The results indicated that 500 mmol/L NaCI treatment could lead to specific features of PCD in root tips, such as DNA ladder, nuclear condense and deformation, and transferase mediated dUTP nick end labeling positive reaction, which were initiated at 4 h of treatment and pro- gressed thereafter. Cytochrome c release from mitochondria into cytoplasm was also observed, which occurred at 2 h and was earlier than the above nuclear events. In the very early phase of PCD, an immediate burst in hydrogen peroxide and superoxide anion production rate was accompanied by two-phase changes of superoxide dismutases and ascorbate peroxidase. A short period of increase in the activity was followed by prolonged impairment. Thus, we conclude that salt can induce PCD in rice root tip cells, and propose that in the early phase of rice root tip cell PCD, salt stress-induced oxidative burst increased the antioxidant enzyme activity, which, in turn, scavenged the ROS and abrogated PCD. Also, when the stress is prolonged, the antioxidant system is damaged and accumulated ROS induces the PCD process, which leads to cytochrome c release and nuclear change.     

等渗Ca(NO3)2和NaCl胁迫对黄瓜幼苗生长及渗透调节物质含量的影响

采用营养液培养方法,以耐盐性较弱的‘津春2号’黄瓜品种为试材,研究了等渗Ca(NO3)2和NaCl胁迫对黄瓜幼苗生长、根系电解质渗透率、根系活力、Na+和K+含量及渗透调节物质含量的影响。结果显示:(1)在84mmol.L-1 NaCl和56mmol.L-1 Ca(NO3)2等渗胁迫下,黄瓜幼苗鲜重和干重均显著下降,且NaCl处理下降的幅度大于等渗Ca(NO3)2处理。(2)NaCl主要通过对黄瓜根系的伤害来抑制植株生长,表现为根系活力下降、根系质膜透性增大、Na+大量积累、K+含量显著下降、Na+/K+明显上升,最终导致根冠比下降;而Ca(NO3)2处理对根系质膜透性、K+含量、Na+/K+的影响均小于NaCl胁迫,且根系活力和根冠比上升,但Ca(NO3)2胁迫后叶片含水量和渗透调节能力均小于NaCl胁迫。(3)NaCl胁迫条件下,黄瓜幼苗内渗透调节物质以可溶性糖为主,而Ca(NO3)2胁迫以可溶性蛋白为主。研究表明,NaCl胁迫对黄瓜幼苗的伤害大于等渗Ca(NO3)2,NaCl主要通过破坏根系质膜结构影响植株生长,而Ca(NO3)2主要通过引起地上部生理干旱来影响植株生长。