边缘细胞对荞麦根尖铝毒的防护效应和对细胞壁多糖的影响

以2个荞麦(Fygopyrum esculentum Moench)基因型‘江西荞麦’(耐性)和‘内蒙荞麦’(敏感)为材料,采用悬空培养(保持边缘细胞附着于根尖和去除根尖边缘细胞),研究边缘细胞对根尖铝毒的防护效应以及对细胞壁多糖组分的影响。结果表明,铝毒抑制荞麦根系伸长,导致根尖Al积累。去除边缘细胞的根伸长抑制率和根尖Al含量高于保留边缘细胞的根。去除边缘细胞使江西荞麦和内蒙荞麦根尖的酸性磷酸酶(APA)活性显著升高,前者在铝毒下增幅更大。同时,铝毒胁迫下去除边缘细胞的根尖果胶甲酯酶(PME)活性和细胞壁果胶、半纤维素1、半纤维素2含量显著高于保留边缘细胞的酶活性和细胞壁多糖含量。表明边缘细胞对荞麦根尖的防护效应,与其阻止Al的吸收,降低根尖细胞壁多糖含量及提高酸性磷酸酶活性有关,以此缓解Al对根伸长的抑制。     

大豆（Glycine max L.）边缘细胞对铝毒的生理生态响应

以大豆浙春2号(铝耐性)和华春18号(铝敏感)为材料,研究了铝胁迫下附着于根尖边缘细胞(即原位边缘细胞)的存活率、根伸长抑制率和PME活性变化以及Al3 对离体后边缘细胞存活率、黏液层厚度的影响。结果表明:铝胁迫下附着于根尖的边缘细胞比离体边缘细胞有更高的活性,前者Al3 处理24h后,其成活率仍能达到74%以上,而后者Al3 处理12h,浙春2号和华春18号边缘细胞的活性在400μmol/L时分别只有44.58%和26.16%;前后两者细胞活性都有随着Al3 浓度升高和处理时间的延长,边缘细胞活性呈越来越低的变化趋势,而离体边缘细胞Al3 处理6h时,相对于敏感性品种而言,高浓度Al3 (≥200μmol/L)有利于铝耐性品种的边缘细胞存活。随Al3 浓度的提升,果胶甲基酯酶(PME)活性增加,根伸长受抑加剧,敏感品种PME活性及铝造成的根伸长抑制均高于耐性品种;同时,Al3 对黏液的产生有一定的影响,黏液层的厚度随Al3 浓度成正向变化趋势。不同Al3 浓度及处理时间下,耐性品种都比敏感品种边缘细胞有较高的活性、分泌较多的黏液。以上结果说明Al3 对边缘细胞具有一定的毒害效应,果胶甲基酯化程度、根伸长受抑及边缘细胞黏液的分泌是根冠对Al毒胁迫反应的结果。     

麻疯树根边缘细胞生物学特性及镉对其活性的影响

以麻疯树为试材,采用悬空气法,探讨麻疯树离体根边缘细胞的生物学特性及重金属镉(Cd2+)对根边缘细胞活性的影响.结果表明,麻疯树根边缘细胞大多数呈椭圆型,少数成弯曲的长条型;根长为15 mm时,边缘细胞的生物学活性(存活率)最大;根冠果胶甲基脂酶(PME)酶活性随着根的伸长有所降低.镉对麻疯树根边缘细胞产生明显毒害,随着镉浓度和处理时间的增加,边缘细胞的活性都呈现下降的趋势.     

四种常见杂草根系及根边缘细胞对铝胁迫的响应

以2种禾本科杂草(升马唐、稗草)和2种菊科杂草(旱莲草、野茼蒿)为实验材料,通过砂培法研究不同科属杂草根部对铝胁迫的响应.结果表明:4种杂草根边缘细胞活性均随着铝胁迫浓度和时间呈显著下降的趋势,但禾本科杂草根系边缘细胞的活性高于菊科杂草,且活性的降低幅度较小;4种杂草根相对伸长率均随铝浓度和处理时间的增加呈递减趋势,但铝对旱莲草和野茼蒿根生长的抑制程度要明显高于升马唐和稗草;根系的铝含量、游离脯氨酸含量、MDA 含量和质膜透性均随铝处理浓度和处理时间的增加而增大,且在高铝浓度(1000 mg · L~(-1))时达到最大值,但升马唐和稗草根系的铝含量、游离脯氨酸含量、MDA含量和质膜透性均显著低于旱莲草和野茼蒿,且随着铝浓度的增加,禾本科杂草根系的游离脯氨酸含量及MDA含量的变化没有达到显著水平(P＞0.05).由此说明,铝毒对杂草造成的伤害随着浓度增加和时间延长而加重;升马唐和稗草的根系通过较高的根边缘细胞活性和根相对伸长率及较低的铝含量、游离脯氨酸含量、MDA含量和质膜透性来增加其对铝的耐性;2种禾本科杂草(升马唐、稗草)的耐铝性高于2种菊科杂草(旱莲草、野茼蒿).     

水稻边缘细胞的生物学特性及对铁毒的响应

以水稻为实验材料,从19个品种中筛选出来相对耐亚铁的水稻品种中优9288和敏感品种汕优10号,研究水稻根边缘细胞的生物学特性及对铁毒的响应.结果表明,第1个水稻边缘细胞几乎与初生根同时出现,当根长为25 mm时,边缘细胞数目最大,分别为1165个(中优9288)、1311个(汕优10号);两个品种边缘细胞活性随着根长的增加而增强,在根长20 mm时活性最强,随着根的进一步伸长,活性下降;根长为2 mm时根冠果胶甲基脂酶(PME)活性最高.随着亚铁溶液浓度的增加,中优9288的边缘细胞数目先增加后减少,当浓度为100 mg/L时达到最大值,汕优10号的边缘细胞数则一直呈现减少趋势;两个品种边缘细胞活性随亚铁溶液浓度增加而降低,当亚铁浓度增加到400 mg/L时,活性有所回升,但当浓度增加到800 mg/L后存活率又下降;根冠PME活性随着亚铁溶液浓度的增加先上升后下降.有关铁毒影响水稻根冠PME活性的机理进行了讨论.     

绿豆根边缘细胞发育特性

以东北绿豆为试验材料,采用琼脂悬空培养法,研究了绿豆边缘细胞的发育特性。结果表明:绿豆根尖发育初期根边缘细胞呈球形,随着根尖伸长逐渐发育形成椭圆形、长椭圆形和长条形;发育过程中,根边缘细胞具有较高的存活率,在根长大于10mm后根边缘细胞的存活率均在70%～80%之间并趋于稳定;在根长为25～30mm时根边缘细胞数目达到最大值(约13 000个);根冠果胶甲基酯酶(PME)活性在根长5mm时达到最高值(1.486H+μmol.root cap-1.h-1),此后随着根的伸长,根冠PME活性在1.107～1.256H+μmol.root cap-1.h-1间变化并趋于稳定。     

高Fe2+ 对水稻离体根边缘细胞的影响

以水稻"汕优10号"为试验材料,悬空气培法获得根边缘细胞,研究水稻边缘细胞的数量、活性和脱离根冠后对高Fe2 的响应过程.结果显示,水稻根长为1 mm时,就形成了205个边缘细胞,随着根的伸长数目不断增加,根长25 mm时,边缘细胞数目最大;根长20 mm时边缘细胞活性最大;根长2 mm时,根冠果胶甲基酯酶(Pectin methyl esterase, PME)活性最高,水稻边缘细胞的形成与根冠PME活性相关.高Fe2 对离体边缘细胞有毒害效应,用浓度为200 μmol·L-1的Fe2 溶液处理48 h后,细胞活性比对照下降了72.70%, 但用400 μmol·L-1 Fe2 溶液处理时,细胞活性有所回升,这可能是边缘细胞耐铁毒的一种应激性反应;根冠PME活性随着Fe2 浓度的增加先上升后下降,表明根冠PME活性与植物铁毒有关.     

高等植物根边缘细胞的发育调控及其生物学功能

植物根边缘细胞是从根冠表皮游离出来并聚集在根尖周围的一群特殊细胞 ,以前曾称为根冠脱落细胞。最近的证据表明 ,绝大多数物种边缘细胞具有生物学活性 ,其发育是受内外信号调控。边缘细胞一旦从根表皮游离后 ,其代谢活性大大上升、基因表达明显不同于根冠细胞。最近 ,与边缘细胞发育早期和晚期相关的两个基因PsUGT1和RCPME1分别被克隆和鉴定。边缘细胞能特异性地合成、分泌一系列的化学物质 ,包括花色素苷、抗生素、特异性酶类及其他化学物质能抑制或促进根际周围的细菌、真菌、病毒、线虫等的生长以及中和根际周围一些有毒化学物质如铝毒。因此 ,边缘细胞在植物生长发育过程中起着多种生物学功能     

高等植物根边缘细胞的发育调控及其生物学功能

植物根边缘细胞是从根冠表皮游离出来并聚集在根尖周围的一群特殊细胞,以前曾称为根冠脱落细胞.最近的证据表明,绝大多数物种边缘细胞具有生物学活性,其发育是受内外信号调控.边缘细胞一旦从根表皮游离后,其代谢活性大大上升、基因表达明显不同于根冠细胞.最近,与边缘细胞发育早期和晚期相关的两个基因PsUGT1和RCPME1分别被克隆和鉴定.边缘细胞能特异性地合成、分泌一系列的化学物质,包括花色素苷、抗生素、特异性酶类及其他化学物质能抑制或促进根际周围的细菌、真菌、病毒、线虫等的生长以及中和根际周围一些有毒化学物质如铝毒.因此,边缘细胞在植物生长发育过程中起着多种生物学功能.     

NaCl胁迫对黄瓜根系边缘细胞发生的影响

以黄瓜为试材,研究了NaCl处理对植株生物量、根长、根系活力、根边缘细胞的数目和活性及黏胶层厚度的影响。结果表明,NaCl处理降低了植株生物量与根系长度,增加了黄瓜幼苗的根系活力。黄瓜边缘细胞的出现几乎与根同时发生,当根长达到25mm时,边缘细胞的数目与活性均达到最大值。NaCl处理对边缘细胞的数目与活性表现出一定的抑制作用。离体根尖的边缘细胞活性也随NaCl处理浓度与处理时间的增加而逐渐减小,但根边缘细胞黏胶层厚度却随NaCl处理浓度的增加而增加。总之,NaCl对黄瓜幼苗造成一定伤害,但根系边缘细胞可通过降解死亡与增加黏胶的分泌量在一定程度上减轻这种伤害程度。     

Response and tolerance of root border cells to aluminum toxicity in soybean seedlings

Root border cells (RBCs) and their secreted mucilage are suggested to participate in the resistance against toxic metal cations, including aluminum (Al), in the rhizosphere. However, the mechanisms by which the individual cell populations respond to Al and their role in Al resistance still remain unclear. In this research, the response and tolerance of RBCs to Al toxicity were investigated in the root tips of two soybean cultivars [Zhechun No. 2 (Al-tolerant cultivar) and Huachun No. 18 (Al-sensitive cultivar)]. Al inhibited root elongation and increased pectin methylesterase (PME) activity in the root tip. Removal of RBCs from the root tips resulted in a more severe inhibition of root elongation, especially in Huachun No. 18. Increasing Al levels and treatment time decreased the relative percent viability of RBCs in situ and in vitro in both soybean cultivars. Al application significantly increased mucilage layer thickness around the detached RBCs of both cultivars. Additionally, a significantly higher relative percent cell viability of attached and detached RBCs and thicker mucilage layers were observed in Zhechun No. 2. The higher viability of attached and detached RBCs, as well as the thickening of the mucilage layer in separated RBCs, suggest that RBCs play an important role in protecting root apices from Al toxicity.     

Interaction between iron plaque and root border cells ameliorates aluminum toxicity of Oryza sativa differing in aluminum tolerance

Root border cells (RBCs), which are generated during plant growth and surround the root cap, and iron plaque (IP), ubiquitously formed on the root surfaces of rice, are known to alleviate aluminum (Al) toxicity. To verify the interactive effects of IP and RBCs on ameliorating Al toxicity, two rice cultivars differing in Al resistance were used to compare Al tolerance between cultivars. Additionally, root elongation, Al uptake and RBCs viability were measured as indicators of the effects of Al. The amounts of DCB-extractable Fe and Al on the root surfaces were much higher in the presence of IP than the absence. IP presence significantly decreased Al-induced inhibition of root elongation and Al contents in roots and root tips. The removal of RBCs from the root tips caused a more severe inhibition of root elongation and a higher Al accumulation in rice roots and root tips. Furthermore, root growth inhibition and Al contents in roots and root tips were significantly lower in roots with a combination of IP and RBCs than in roots with IP or RBCs only. The formation of IP on the root surface maintained higher RBCs viability and depressed mucilage exudation in an Al-tolerant rice cultivar. The results suggest that both IP and RBCs ameliorate Al toxicity, and IP has a greater capacity for Al resistance. The combination of IP and RBCs exhibited a synergistic effect associated with Al resistance.     

Colonization of Tomato Root Seedling by Pseudomonas fluorescens 92rkG5: Spatio–temporal Dynamics, Localization, Organization, Viability, and Culturability

The localization, viability, and culturability of Pseudomonas fluorescens 92rkG5 were analyzed on three morphological root zones (root tip + elongation, root hair, and collar) of 3-, 5-, and 7-day-old tomato plants. Qualitative information about the localization and viability was collected by confocal laser scanning microscopy. Quantitative data concerning the distribution, viability, and culturability were obtained through combined dilution plating and flow cytometry. Colonization by P. fluorescens affected root development in a complex way, causing a general increase in the length of the collar and early stimulation of the primary root growth (3rd day), followed by a reduction in length (7th day). The three root zones showed different distribution, organization, and viability of the bacterial cells, but the distribution pattern within each zone did not change with time. Root tips were always devoid of bacteria, whereas with increasing distance from the apex, microcolonies or strings of cells became more and more prominent. Viability was high in the elongation zone, but it declined in the older parts of the roots. The so-called viable but not culturable cells were observed on the root, and their proportion in the distal (root tip + elongation) zone dramatically increased with time. These results suggest the existence of a specific temporal and spatial pattern of root colonization, related to cell viability and culturability, expressed by the plant-beneficial strain P. fluorescens 92rkG5.     

Possible role of root border cells in detection and avoidance of aluminum toxicity

Root border cells are living cells that surround root apices of most plant species and are involved in production of root exudates. We tested predictions of the hypothesis that they participate in detection and avoidance of aluminum (Al) toxicity by comparing responses of two snapbean (Phaseolus vulgaris) cultivars (cv Dade and cv Romano) known to differ in Al resistance at the whole-root level. Root border cells of these cultivars were killed by excess Al in agarose gels or in simple salt solutions. Percent viability of Al-sensitive cv Romano border cells exposed in situ for 96 h to 200 microM total Al in an agarose gel was significantly less than that of cv Dade border cells; similarly, relative viability of harvested cv Romano border cells was significantly less than that of cv Dade cells after 24 h in 25 microM total Al in a simple salt solution. These results indicate that Al-resistance mechanisms that operate at the level of whole roots also operate at the cellular level in border cells. Al induced a thicker mucilage layer around detached border cells of both cultivars. Cultivar Dade border cells produced a thicker mucilage layer in response to 25 microM Al compared with that of cv Romano cells after 8 h of treatment and this phenomenon preceded that of observed cultivar differences in relative cell viability. Release of an Al-binding mucilage by border cells could play a role in protecting root tips from Al-induced cellular damage.     

Effect of toxic Fe2+ levels on the biological characteristics of rice root border cells

Root border cells are a population of rhizosphere cells surrounding the root tips but separated from them. The root tip is a major target of Fe²⁺ toxicity; thus, it was hypothesized that the border cells might protect or exacerbate Fe²⁺ toxicity. To explore the effects of excess Fe²⁺ on the border cells in rice (Oryza sativa L.), experiments were carried out using the border cells in vitro (Shanyou No. 10). The border cells were precultured under ??hanging in the air?? and detached from the root tips. The shape, numbers, and viability of border cells were examined during exposure to toxic levels of Fe²⁺. When the root was 1 mm long, there were 205 border cells on average. With the growth of the root, more border cells were observed. When the root grew to 25 mm long, the total number of border cells reached a maximum, while the maximum activity of border cells appeared when the root was 20 mm long. The pectin methyl esterase (PME) activity of the root cap peaked at a root length of 2 mm. Border cell development was related to PME activity in rice. Excessive Fe²⁺ was toxic to detached border cells. After treatment with 200 ??M Fe²⁺ solution for 48 h, cell viability decreased by 72.70%. However, when treated with 400 ??M Fe²⁺ solution, the number of viable cells was actually higher, suggesting the induction of a cellular self-protection response. The activity of PME first increased under high concentrations of Fe²⁺ and then decreased. These results indicate that toxic levels of Fe²⁺ modulate PME activity and border cell survival.     

Elongation of barley roots in high‐pH nutrient solution is supported by both cell proliferation and differentiation in the root apex

Many crops grow well on neutral or weakly acidic soils. The ability of roots to elongate under high‐external pH would be advantageous for the survival of plants on alkaline soil. We found that root elongation was promoted in some plant species in alkaline‐nutrient solution. Barley, but not tomato, root growth was maintained in pH 8 nutrient solution. Fe and Mn were absorbed well from the pH 8 nutrient solution by both barley and tomato plants, suggesting that the different growth responses of these two species may not be caused by insolubilization of transition metals. The ability of intact barley and tomato plants to acidify external solution was comparable; in both species, this ability decreased in plants exposed to pH 8 nutrient solution for 1 w. Conversely, cell proliferation and elongation in barley root apices were facilitated at pH 8 as shown by microscopy and cell‐cycle‐related gene‐expression data; this was not observed in tomato. We propose that barley adapts to alkaline stress by increasing root development.     

Aluminum targets elongating cells by reducing cell wall extensibility in wheat roots

Phytotoxicity of aluminum is characterized by a rapid inhibition of root elongation at micromolar concentrations, however, the mechanisms primarily responsible for this response are not well understood. We investigated the effect of Al on the viscosity and elasticity parameters of root cell wall by a creep-extension analysis in two cultivars of wheat (Triticum aestivum L.) differing in Al resistance. The root elongation and both viscous and elastic extensibility of cell wall of the root apices were hardly affected by the exposure to 10 microM Al in an Al-resistant cultivar, Atlas 66. However, similar exposure rapidly inhibited root elongation in an Al-sensitive cultivar, Scout 66 and this was associated with a time-dependent accumulation of Al in the root tissues with more than 77% residing in the cell wall. Al caused a significant decrease in both the viscous and elastic extensibility of cell wall of the root apices of Scout 66. The "break load" of the root apex of Scout 66 was also decreased by Al. However, neither the viscosity nor elasticity of the cell wall was affected by in vitro Al treatment. Furthermore, pre-treatment of seedlings with Al in conditions where root elongation was slow (i.e. low temperature) did not affect the subsequent elongation of roots in a 0 Al treatment at room temperature. These results suggest that the Al-dependent changes in the cell wall viscosity and elasticity are involved in the inhibition of root growth. Furthermore, for Al to reduce cell wall extensibility it must interact with the cell walls of actively elongating cells.     

Al3+对大豆根边缘细胞程序性死亡诱导的生理生态作用

 设置不同的Al ³⁺浓度（0、25、50、100、200、400 μmol·L^-1）和培养时间 （12、24 h）,研究了边缘细胞活性和大豆（Glycine max）根中 过氧化氢酶（CAT）、过氧化物酶POD）、超氧化物歧化酶SOD）随Al ³⁺浓度及处理时间变化的规律,并通过Hoec hst333 42-PI双重荧光染色、 梯状DNA（即DNA ladder）分析和末端脱氧核糖核酸转移酶介导的dUTP切口末端标记（即TUNEL原位标记）检测,研究了Al ³⁺对大豆根边缘细胞 程序性死亡诱导的生理生态作用。结果表明,Al ³⁺胁迫能诱导边缘细胞的死亡,随着Al ³⁺浓度的升高和处理时间的延长,细胞死亡率增加。通 过Hoechst33342-PI双重荧光染色、DNA ladder分析和TUNEL原位标记,检测到Al ³⁺胁迫下发生程序性死亡的边缘细胞。其表现为：在 400μmol·L^-1 Al ³⁺诱导大豆根24 h时, 核酸电泳显示细胞DNA发生特异性降解并形成阶梯状电泳条带(DNA ladder),用TUNEL原位标记检测200 和400μmol·L^-1 Al ³⁺处理12 h后的大豆根 边缘细胞,发现DNA的3′-OH端被原位特异标记,二氨基联苯胺（DAB）显色后,细胞核为阳性或强 阳性。同时,高浓度Al ³⁺ （＞100μmol·L^-1）处理下,CAT、POD和S OD活性均有不同程度的下降,CAT和SOD的活性也随处理时间的延长而降低 。说明在Al ³⁺胁迫下边缘细胞的死亡可能是一种程序性死亡形式,高浓度Al ³⁺胁迫下,通过诱导活性氧在细胞体内的产生和累积而导致细胞凋 亡,此过程是其对逆境胁迫所作出的生理生态防御性应答方式之一。