茉莉酸甲酯和水杨酸对丹参幼苗中蔗糖代谢和酚酸类物质积累的影响

以丹参幼苗为材料,研究了茉莉酸甲酯(MeJA)和水杨酸(SA)对其地上地下部分蔗糖代谢和根系中酚酸类物质积累的影响.结果显示:(1)外源施用MeJA能够显著增强幼苗叶片和根中酸性蔗糖转化酶活性,促进蔗糖降解,减少根系中蔗糖含量;同时根中丹参素(来源于酪氨酸-衍生物支路)、原儿茶酸(类苯丙烷支路的中间产物)、咖啡酸(来源于类苯丙烷支路)、迷迭香酸(来源于类苯丙烷支路和酪氨酸-衍生物支路)、丹酚酸B(丹参素、咖啡酸、迷迭香酸的衍生物)以及总酚酸含量均显著增加.(2)外施SA可显著降低幼苗叶片和根中酸性蔗糖转化酶活性及叶片中性蔗糖转化酶活性,抑制蔗糖降解,显著增加地上部分蔗糖含量,根中蔗糖含量和对照相比差异不显著;同时根中丹参素含量减少,但原儿茶酸、咖啡酸以及迷迭香酸含量增加,丹酚酸B和总酚酸类物质含量与对照相比差异不显著.因此推测,植物中蔗糖代谢影响丹参素合成的酪氨酸-衍生物支路,而不影响原儿茶酸、咖啡酸及迷迭香酸等合成的类苯丙烷支路.     

外源NO对铝毒下水稻根系生长和抗氧化系统的影响

以水稻品种‘浙辐802’为材料,采用水培法研究铝毒下外源NO对幼苗根系生长、活性氧产生和抗氧化酶活性的影响,探讨外源NO提高水稻耐铝性的生理生化机制。结果显示:(1)0.05mmol/L Al显著抑制水稻根系生长,促使根尖Al、胼胝质、过氧化氢(H2O2)和超氧阴离子自由基(O-·2)含量显著增加;而外源0.1mmol/L的NO供体亚硝基铁氰化钠(sodium nitroprusside,SNP)预处理能使铝毒下水稻幼苗根相对伸长率及根尖NO含量分别增加34.96%和12.86%,根尖Al和相对胼胝质含量分别下降83.04%和31.93%,表明NO可部分缓解铝毒害,且这种作用与内源NO含量变化有关。(2)外源NO同时使铝毒下水稻幼苗根尖H2O2和O-·2含量分别下降15.43%和12.93%,使超氧化物歧化酶(SOD)、过氧化物酶(POD)和过氧化氢酶(CAT)活性显著上升,且外源NO的该种作用可以被0.075mmol/L NO清除剂(carboxy-PTIO,cPTIO)所逆转。研究表明,外源NO在调节活性氧代谢以维持细胞膜结构稳定,进而有效减轻Al对水稻根系的损伤上起着重要作用。     

干旱胁迫下外源茉莉酸甲酯对玉米幼苗根系吸水的影响

茉莉酸类化合物作为环境信号分子,不仅参与植物生长发育的调控,同时受到环境胁迫的诱导,参与植物对逆境胁迫的响应和防御。本研究以北方广泛种植的玉米品种‘郑单958’为材料,通过对根系外源施加茉莉酸甲酯的方式,探究干旱胁迫下茉莉酸甲酯对玉米幼苗抗旱性以及根系吸水的影响。结果表明,外源茉莉酸甲酯可提高玉米幼苗光合速率、蒸腾作用和气孔导度,增强抗氧化酶活性,降低H_2O_2和丙二醛的含量,从而缓解干旱胁迫对植株造成的损伤。通过测定根系水导、氯化汞处理的蒸腾速率的变化以及水通道蛋白的表达量,发现干旱胁迫下外源茉莉酸甲酯可增强根系水通道蛋白的表达,进而增强玉米幼苗的根系吸水能力,从而缓解干旱胁迫造成的叶片水分含量的下降和水势的降低,提高了玉米幼苗的抗旱性。     

Cd-K双重处理对水稻幼苗生长和Cd吸收转运的影响及相关转运通道分析

以水稻( Oryza sativa Linn.)高 Cd 积累品种‘T 优705’(‘T You 705’)和低 Cd 积累品种‘湘早籼24’(‘Xiangzaoxian 24’)为实验材料,采用水培法对不同浓度Cd(0.0和2.7μmol·L-1 Cd)和K(0、30和60 mmol·L-1 K)处理条件下2个品种幼苗的相对生长量、根系和地上部的Cd含量及其亚细胞分布特征进行了比较,并分析了添加离子通道活性抑制剂TEA和LaCl3后幼苗根系和地上部的Cd和K含量;在此基础上,比较了NSCCs(非选择性阳离子通道)和K专性通道对2个品种幼苗根系和地上部Cd和K吸收贡献率的影响。结果表明：与Cd单一处理组(2.7μmol·L-1 Cd)相比, Cd-K双重处理组(2.7μmol·L-1 Cd-30 mmol·L-1 K和2.7μmol·L-1 Cd-60 mmol·L-1 K)2个品种幼苗的相对生长量显著提高,而幼苗根系和地上部的Cd含量显著下降;随K浓度的提高,2个品种幼苗根系细胞壁和细胞液中的Cd含量显著下降,但细胞壁中Cd含量的分配比例增大而细胞液中Cd含量的分配比例则减小。在含2.7μmol·L-1 Cd和30 mmol·L-1 K的培养液中分别添加5 mmol·L-1 TEA或0.2 mmol·L-1 LaCl3后,2个品种幼苗根系和地上部的Cd和K含量均显著下降,其中,LaCl3处理组的根系Cd含量降幅高于TEA处理组,但LaCl3处理组的根系K含量降幅则低于TEA组。 NSCCs对品种‘T优705’幼苗根系和地上部Cd吸收的贡献率显著低于品种‘湘早籼24’幼苗,而K专性通道对品种‘T优705’幼苗根系K吸收和地上部Cd吸收的贡献率则显著低于品种‘湘早籼24’幼苗。研究结果显示：添加外源K可缓解Cd对水稻幼苗生长的抑制作用,并通过提高细胞壁与Cd的结合能力来降低细胞液中Cd的积累,以此减弱幼苗对Cd的吸收和转运能力;幼苗体内的K和Cd均可通过K专性通道和NSCCs转运,其中,K吸收和转运主要通过K专性通道完成,而Cd吸收和转运主要通过NSCCs完成。此外,品种‘T优705’可能具有多种离子通道参与Cd的吸收和转运,而品种‘湘早籼24’主要依赖NSCCs参与Cd的吸收和转运,且后者对K的吸收和积累强于前者。     

外源NO对镉胁迫下水稻幼苗抗氧化系统和微量元素积累的影响

重金属污染已成为世界范围的主要问题之一,其中镉(Cd)毒害的范围最广.Cd污染影响植物的生长和发育.一氧化氮(NO)作为植物体内的一种活性信号因子,参与了植物对各种胁迫的应答.为了探讨外源NO对Cd胁迫下水稻苗期生理生化响应的调节作用,以粳稻和籼稻为材料,采用营养液水培的方法,研究外源NO供体硝普钠(SNP)对100μmol/L Cd胁迫下2个水稻基因型(ZF802和ZH11)幼苗生长、抗氧化酶系统和微量元素吸收的影响.结果表明,1.5 mmol/L SNP能明显缓解Cd胁迫对两种水稻幼苗生长的抑制作用,提高幼苗对Cd的耐性.尽管存在一定的基因型差异,但外源NO能使两个基因型叶片中的超氧化物歧化酶(SOD)和过氧化物酶(POD)活性下降,降低了叶片中丙二醛(MDA)含量,从而缓解Cd对水稻幼苗的毒害.另外,外源NO影响Cd胁迫下水稻幼苗地上部和根部Cd和微量元素的积累,这种影响的程度与水稻的品种有关,具有显著的基因型差异,且机制较复杂.     

基于~(32)P示踪的不同供磷环境杉木幼苗磷的分配规律分析

通过分析杉木(Cunninghamia lanceolata)幼苗磷(P)分配规律,可以阐明两个磷高效利用杉木在不同供磷水平下吸收外源磷的分配及动态变化,为进一步进行磷高效利用基因型的选育提供参考。该研究以2个磷高效利用杉木家系(被动忍受型M1与主动活化型M4)幼苗为试验材料,利用~(32)P同位素示踪技术,研究在不同供磷水平下2个杉木家系幼苗磷分配规律。结果表明,M1和M4吸收的外源磷的含量分布特征均为根叶茎,自显影中相同处理时期的各器官在水平投影面上~(32)P含量均为根茎叶。低磷处理下M1和M4根、茎、叶吸收的外源磷的含量均明显低于高磷处理,自显影中相同处理时间根、茎、叶低磷水平下成像的黑化程度也低于高磷水平,且低磷处理下吸收的外源磷的含量增加缓慢,说明低磷胁迫严重影响杉木苗磷的吸收与积累。M1和M4的根系磷分配率在低磷胁迫下呈现出明显的先减少后增加趋势,高磷水平下根系磷分配率表现为先增加后趋于平稳。这说明M1和M4可以通过体内磷的重新分配来适应外界低磷胁迫,即杉木苗在低磷胁迫初期将根系中的磷转移至地上部分,随着胁迫时间的延长,地上部分的磷向根系中转移。但两个家系在低磷条件下对吸收的外源磷的分配格局差异明显:从开始至结束M1吸收的外源磷的分配率表现为根系地上部分,而M4先表现为根系地上部分,后表现为地上部分根系,说明M1在低磷胁迫后加强体内磷循环的程度相比于M4更高,即磷从地上部分向根系转移的趋势更强烈。     

Phosphorus distribution inside Chinese fir seedlings under different P supplies based on 32P tracer

通过分析杉木(Cunninghamia lanceolata)幼苗磷(P)分配规律, 可以阐明两个磷高效利用杉木在不同供磷水平下吸收外源磷的分配及动态变化, 为进一步进行磷高效利用基因型的选育提供参考。该研究以2个磷高效利用杉木家系(被动忍受型M1与主动活化型M4)幼苗为试验材料, 利用 ³²P同位素示踪技术, 研究在不同供磷水平下2个杉木家系幼苗磷分配规律。结果表明, M1和M4吸收的外源磷的含量分布特征均为根>叶>茎, 自显影中相同处理时期的各器官在水平投影面上 ³²P含量均为根>茎>叶。低磷处理下M1和M4根、茎、叶吸收的外源磷的含量均明显低于高磷处理, 自显影中相同处理时间根、茎、叶低磷水平下成像的黑化程度也低于高磷水平, 且低磷处理下吸收的外源磷的含量增加缓慢, 说明低磷胁迫严重影响杉木苗磷的吸收与积累。M1和M4的根系磷分配率在低磷胁迫下呈现出明显的先减少后增加趋势, 高磷水平下根系磷分配率表现为先增加后趋于平稳。这说明M1和M4可以通过体内磷的重新分配来适应外界低磷胁迫, 即杉木苗在低磷胁迫初期将根系中的磷转移至地上部分, 随着胁迫时间的延长, 地上部分的磷向根系中转移。但两个家系在低磷条件下对吸收的外源磷的分配格局差异明显: 从开始至结束M1吸收的外源磷的分配率表现为根系>地上部分, 而M4先表现为根系>地上部分, 后表现为地上部分>根系, 说明M1在低磷胁迫后加强体内磷循环的程度相比于M4更高, 即磷从地上部分向根系转移的趋势更强烈。     

外源EBR对NaCl胁迫下燕麦幼苗无机离子吸收、运输和分配的影响

为了探讨外源2,4-表油菜素内酯(2,4-epibrassinolide,EBR)诱导燕麦(Avena sativa L.)幼苗抗盐性的效果及其生理调节机制,以"青引2号"和"加燕2号"燕麦为材料,研究NaCl胁迫下施用外源EBR对燕麦幼苗无机离子吸收、运输和分配的影响。结果表明:100mmol·L-1NaCl胁迫下,"青引2号"和"加燕2号"燕麦幼苗叶片和根系中的Na+、Cl-含量均显著升高,对阳离子的吸收产生了拮抗作用,导致燕麦幼苗叶片和根系中的K+、Ca2+、Mg2+、Mn2+、Fe2+、Zn2+、Cu2+含量显著降低,离子稳态平衡被打破;100 mmol·L-1NaCl胁迫下,施用0.01μmol·L-1外源EBR后,"青引2号"和"加燕2号"燕麦幼苗叶片和根系中的Na+和Cl-含量显著降低,促进了燕麦幼苗根系对K+、Ca2+、Mg2+、Fe2+、Mn2+、Cu2+和Zn2+的吸收,叶片和根系中K+/Na+、Cl-/Na+、Ca2+/Na+、Mg2+/Na+、Fe2+/Na+、Mn2+/Na+、Cu2+/Na+和Zn2+/Na+显著升高,并且有效调控燕麦幼苗体内无机离子的运输比和阳离子的运输选择性比率,离子稳态重新达到平衡状态;说明外源EBR能够缓解NaCl胁迫下Na+和Cl-对燕麦幼苗所造成的离子毒害作用,有效调控燕麦幼苗对无机离子的选择性吸收、运输和分配,对维持燕麦幼苗体内的离子稳态平衡具有正向调控作用。     

外源胆固醇对水稻幼苗膜脂组成及抗冷力的影响

分析了水稻幼苗低温胁迫前后膜脂和膜脂脂肪酸含量变化。结果表明,经胆固醇处理的幼苗叶和根细胞膜脂中LPC、PS和PG含量比对照下降少,PA含量增加也较少。胆固醇处理的幼苗叶和根棕榈酸(16:0)增加量和亚麻酸(18:3)与IUFA减少量均明显比对照少。试验结果证明,水稻幼苗叶片和根系的抗冷力与PA含量和脂肪酸不饱和程度变化有密切关系。外源胆固醇处理水稻幼苗能阻止低温对膜脂的破坏作用,提高幼苗抗低温胁迫能力。     

外源EBR对NaCl胁迫下燕麦幼苗无机离子吸收、运输和分配的影响

为了探讨外源2,4-表油菜素内酯(2,4-epibrassinolide,EBR)诱导燕麦(Avena sativa L.)幼苗抗盐性的效果及其生理调节机制,以"青引2号"和"加燕2号"燕麦为材料,研究NaCl胁迫下施用外源EBR对燕麦幼苗无机离子吸收、运输和分配的影响。结果表明:100mmol·L-1NaCl胁迫下,"青引2号"和"加燕2号"燕麦幼苗叶片和根系中的Na+、Cl-含量均显著升高,对阳离子的吸收产生了拮抗作用,导致燕麦幼苗叶片和根系中的K+、Ca2+、Mg2+、Mn2+、Fe2+、Zn2+、Cu2+含量显著降低,离子稳态平衡被打破; 100 mmol·L-1NaCl胁迫下,施用0.01μmol·L-1外源EBR后,"青引2号"和"加燕2号"燕麦幼苗叶片和根系中的Na+和Cl-含量显著降低,促进了燕麦幼苗根系对K+、Ca2+、Mg2+、Fe2+、Mn2+、Cu2+和Zn2+的吸收,叶片和根系中K+/Na+、Cl-/Na+、Ca2+/Na+、Mg2+/Na+、Fe2+/Na+、Mn2+/Na+、Cu2+/Na+和Zn2+/Na+显著升高,并且有效调控燕麦幼苗体内无机离子的运输比和阳离子的运输选择性比率,离子稳态重新达到平衡状态;说明外源EBR能够缓解NaCl胁迫下Na+和Cl-对燕麦幼苗所造成的离子毒害作用,有效调控燕麦幼苗对无机离子的选择性吸收、运输和分配,对维持燕麦幼苗体内的离子稳态平衡具有正向调控作用。     

Abnormal accumulation of sugars and phenolics in tobacco roots expressing the Agrobacterium T-6b oncogene and the role of these compounds in 6b-induced growth

The Agrobacterium T-DNA oncogene 6b induces tumors and modifies the growth of transgenic plants by an unknown mechanism. We have investigated changes in roots of tobacco seedlings that express a dexamethasone-inducible T-6b (dex-T-6b) gene. On induction medium with sucrose, intact or isolated dex-T-6b roots accumulated sucrose, glucose, and fructose and changed their growth, contrary to noninduced roots. Root fragments bridging agar blocks with or without sucrose accumulated sugars at the site of sucrose uptake, resulting in local growth. Induced root fragments showed enhanced uptake of 14C-labeled sucrose, glucose, and fructose. When seedlings were placed on sucrose-free induction medium, sugar levels strongly decreased in roots and increased in cotyledons. Collectively, these results demonstrate that 6b stimulates sugar uptake and retention with drastic effects on growth. Apart from sugars, phenolic compounds also have been found to accumulate in 6b tissues and have been proposed earlier to play a role in 6b-induced growth. Induced dex-T-6b roots accumulated high levels of 5-caffeoylquinic acid (or chlorogenic acid [CGA]), but only under conditions where endogenous sugars increased. Inhibition of phenylalanine ammonia-lyase with the competitive inhibitor 2-aminoindan-2-phosphonic acid (AIP) abolished CGA accumulation without modifying sugar accumulation or affecting the 6b phenotype. We conclude that the absorption, retention, and abnormal accumulation of sugars are essential factors in 6b-induced growth changes, whereas phenylpropanoids only marginally contribute to the 6b seedling phenotype.     

Sugar uptake and transport in rice embryo. Expression of companion cell-specific sucrose transporter (OsSUT1) induced by sugar and light

We investigated sugar uptake and transport in rice (Oryza sativa) embryo during grain germination. Endogenous sugar levels, accumulation of starch granules, and gene expression of a rice sucrose transporter (OsSUT1) were examined using rice embryos germinated with or without exogenous sugar supply. Starch granules remarkably accumulated in the cells around vascular bundles as a consequence of the sugar taken up by the embryos, indicating that the taken-up sugars are transiently converted into starch. In situ detection for OsSUT1 mRNA indicated its localization in the phloem companion cells. Furthermore, northern-blot and in situ hybridization analyses showed that OsSUT1 expression is not detectable in embryos subjected to sugar starvation conditions, whereas its expression is enhanced by an increased endogenous sugar level. Overall results indicate that the expression of companion cell-specific sucrose transporter, OsSUT1 is regulated by the endogenous sugar status as well as light exposure.     

Excess nickel modulates activities of carbohydrate metabolizing enzymes and induces accumulation of sugars by upregulating acid invertase and sucrose synthase in rice seedlings

The effects of increasing concentrations of nickel sulfate, NiSO₄ (200 and 400 μM) in the growth medium on the content of starch and sugars and activity levels of enzymes involved in starch and sugar metabolism were examined in seedlings of the two Indica rice cvs. Malviya-36 and Pant-12. During a 5–20 day growth period of seedlings in sand cultures, with Ni treatment, no definite pattern of alteration in starch level could be observed in the seedlings. In both roots and shoots of the seedlings Ni treatment led to a significant decrease in activities of starch degrading enzymes α-amylase, β-amylase, whereas starch phosphorylase activity increased. The contents of reducing, non-reducing, and total sugars increased in Ni-treated rice seedlings with a concomitant increase in the activities of sucrose degrading enzymes acid invertase and sucrose synthase. However, the activity of sucrose synthesizing enzyme sucrose phosphate synthase declined. These results suggest that Ni toxicity in rice seedlings causes marked perturbation in metabolism of carbohydrates leading to increased accumulation of soluble sugars. Such perturbation could serve as a limiting factor for growth of rice seedlings in Ni polluted environments and accumulating soluble sugars could serve as compatible solutes in the cells under Ni toxicity conditions.     

Tissue and cell-specific localization of rice aquaporins and their water transport activities

Water transport in plants is greatly dependent on the expression and activity of water transport channels, called aquaporins. Here, we have clarified the tissue- and cell-specific localization of aquaporins in rice plants by immunoblotting and immunocytochemistry using seven isoform-specific aquaporin antibodies. We also examined water transport activities of typical aquaporin family members using a yeast expression system in combination with a stopped-flow spectrophotometry assay. OsPIP1 members, OsPIP2;1, OsTIP1;1 and OsTIP2;2 were expressed in both leaf blades and roots, while OsPIP2;3, OsPIP2;5 and OsTIP2;1 were expressed only in roots. In roots, large amounts of aquaporins accumulated in the region adjacent to the root tip (around 1.5-4 mm from the root tip). In this region, cell-specific localization of the various aquaporin members was observed. OsPIP1 members and OsTIP2;2 accumulated predominantly in the endodermis and the central cylinder, respectively. OsTIP1;1 showed specific localization in the rhizodermis and exodermis. OsPIP2;1, OsPIP2;3 and OsPIP2;5 accumulated in all root cells, but they showed higher levels of accumulation in endodermis than other cells. In the region at 35 mm from the root tip, where aerenchyma develops, aquaporins accumulated at low levels. In leaf blades, OsPIP1 members and OsPIP2;1 were localized mainly in mesophyll cells. OsPIP2;1, OsPIP2;3, OsPIP2;5 and OsTIP2;2 expressed in yeast showed high water transport activities. These results suggest that rice aquaporins with various water transport activities may play distinct roles in facilitating water flux and maintaining the water potential in different tissues and cells.     

Monosaccharide/proton symporter AtSTP1 plays a major role in uptake and response of Arabidopsis seeds and seedlings to sugars

The aim of this study was to investigate the in vivo properties and function of the high-affinity monosaccharide/proton symporter AtSTP1 of Arabidopsis. We isolated an Atstp1 knock-out mutant and found that this plant grows and develops normally. The AtSTP1 gene is expressed in germinating seeds and seedlings, with AtSTP1 activity found mainly in the seedling root. The rate of uptake of [(14)C]-3-O-methylglucose and [(14)C]-D-glucose is 60% less in Atstp1 seedlings than in the wild type, showing that AtSTP1 is the major monosaccharide transporter in Arabidopsis seedlings. Transport of D-galactose and D-mannose is also up to 60% less in Atstp1 seedlings compared to wild type, but transport of D-fructose, L-arabinose and sucrose is not reduced. Germination of Atstp1 seed shows reduced sensitivity to D-mannose, demonstrating that AtSTP1 is active before germination. Atstp1 seedlings grow effectively on concentrations of D-galactose that inhibit wild-type growth, even at up to 100 mM D-galactose, indicating that active transport by AtSTP1 plays a major role at very high concentrations of exogenous sugar. These findings provide insight into the physiological function of AtSTP1 and clearly establish its importance in the uptake of extracellular sugars by the embryo and in seedlings.     

Rice calcium‐dependent protein kinase OsCPK17 targets plasma membrane intrinsic protein and sucrose‐phosphate synthase and is required for a proper cold stress response

Calcium‐dependent protein kinases (CDPKs) are involved in plant tolerance mechanisms to abiotic stresses. Although CDPKs are recognized as key messengers in signal transduction, the specific role of most members of this family remains unknown. Here, we test the hypothesis that OsCPK17 plays a role in rice cold stress response by analysing OsCPK17 knockout, silencing and overexpressing rice lines under low temperature. Altered OsCPK17 gene expression compromises cold tolerance performance, without affecting the expression of key cold stress‐inducible genes. A comparative phosphoproteomic approach led to the identification of six potential in vivo OsCPK17 targets, which are associated with sugar and nitrogen metabolism, and with osmotic regulation. To test direct interaction, in vitro kinase assays were performed, showing that the sucrose‐phosphate synthase OsSPS4 and the aquaporin OsPIP2;1/OsPIP2;6 are phosphorylated by OsCPK17 in a calcium‐dependent manner. Altogether, our data indicates that OsCPK17 is required for a proper cold stress response in rice, likely affecting the activity of membrane channels and sugar metabolism.     

Bioregulator enhancement of sink activity in sugar beet

The sink mobilizing abillity is partially determined by sugar uptake rates of storage cells. Two synthetic growth regulators (Pix and BAS 106W) were tested for their effect on sucrose uptake in root tissue discs or glucose uptake in cell cultures of sugar beet. In tissue discs, uptake at the plasmalemma was determined by incubating the discs for 1 h in the presence of 5 mM sucrose and at the tonoplast for 4 h in the presence of 40 mM sucrose. Cell cultures were incubated for 1 h in the presence of 1 mM glucose. Pix (10 mg l^–1) caused a 20% stimulation of active sucrose uptake at the plasmalemma. Active sucrose uptake at the tonoplast was increased 67% by 100 mg l^–1 Pix. No effect of BAS 106W was observed on sucrose uptake in tissue discs. In cell cultures, a 65% enhancement of active glucose uptake was observed with both Pix and BAs 106W. When the bioregulators were applied to the root medium of seedlings, Pix but not BAS 106W resulted in increased root/shoot ratio, translocation of ¹⁴C-assimilates, and allocation of more biomass to the root sink. The data suggested that sugar transport and translocation may be used as biochemical criteria for rapid screening of effective yield enhancing bioregulators.