镉在东南景天中的亚细胞分配

采用差速离心技术,比较研究了Cd在矿山生态型东南景天(Sedum alfredii Hance)根、茎、叶中的亚细胞分布。结果表明:矿山生态型东南景天对Cd具有很好的忍受耐和累积能力,非矿山生态型东南景天则不具有这种能力。Cd在矿山生态型东南景天根、茎、叶各部分的亚细胞分配满足F1(细胞壁部分)>F3(可溶部分)>F2(细胞器及膜)规律,且Cd在细胞壁部分的分配占绝对优势;同时矿山生态型东南景天地上部有很好的Cd累积能力。与众多超累积植物相似的Cd亚细胞分配规律和地上部的良好的Cd累积能力。对此植物的Cd亚细胞分布完全不同于非矿山生态型东南景天的情况作了比较分析,结果显示矿山生态型东南景天很可能是一种新的镉超累积种质资源。     

古老铅锌矿山生态型东南景天对锌耐性及超积累特性的研究

植物长期生长在重金属污染的生境中,逐渐进化成不同的生态型.通过调查中国东南部古老Pb/Zn矿和非矿山生境中的植物种群,发现生长在古老Pb/Zn矿的东南景天(Sedum alfredii Hance)是一种新的Zn超积累植物.在自然和控制条件下,古老Pb/Zn矿生态型比非矿山生态型植株的茎粗、叶片大、植株高.在矿山土壤Zn有效含量为105.5～325.4 mg·kg-1时,矿山生态型东南景天植株地上部Zn含量为4134～5000 mg·     

超积累和非超积累生态型东南景天茎、叶内生细菌多样性分析

【目的】探究和比较超积累和非超积累生态型东南景天茎、叶微生物群落结构的异同。【方法】采用高通量测序技术研究野外两种生态型东南景天茎和叶片的内生细菌群落结构。【结果】4个样品总共得到366 783条有效序列和39 948个OTU(97%相似度)。从Shannon指数得知:两种生态型东南景天叶片内生菌的多样性均高于茎;超积累生态型东南景天叶片内生菌的多样性高于非超积累生态型,但非超积累生态型东南景天茎组织中内生菌多样性高于超积累生态型东南景天。超积累生态型东南景天的叶片和茎中的内生菌分别包括26和21个门,123和117个科;非超积累生态型东南景天叶片和茎中的内生菌分别包括43和22个门,175和83个科,4个样品的优势菌群均为变形菌门(Proteobacteria)、拟杆菌门(Bacteroidetes)、厚壁菌门(Firmicutes)、放线菌门(Actinobacteria)和蓝藻细菌门(Cyanobacteria)。在属分类水平上,超累积生态型东南景天叶片和茎第一优势菌属分别为Synechococcus和Plesiomonas;非超积累生态型东南景天叶片和茎组织第一优势菌群分别为Pseudomonas和Dechloromonas。【结论】两种生态型东南景天的叶片和茎中均具有丰富的内生细菌,但超积累生态型东南景天叶片内生菌多样性最大,且存在一些独有的功能菌群。     

不同生态型东南景天对土壤中Cd的生长反应及吸收积累的差异性

 采用盆栽方法研究了两种生态型东南景天(Sedum alfredii)对土壤中不同含量Cd(即对照, 12.5, 25, 50, 100, 200, 300, 400 mg&;#8226;kg^－1)的生 长反应、吸收和积累Cd的差异性。结果表明,土壤添加重金属Cd后,矿山生态型东南景天生长正常,地上部和根系Cd含量随着土壤中Cd含量的 增加而增加,在400 mg&;#8226;kg^－1 Cd处理下含量分别高达2 900和500 mg&;#8226;kg^－1,其地上部显著大于根部;然而,土壤添加Cd后,非矿山生态型东 南景天的生长受到抑制,地上部和根部的生物量显著降低。当土壤Cd含量为50～100 mg&;#8226;kg^－1 时,非矿山生态型东南景天的地上部和根系Cd含 量随着土壤中Cd含量的增加而增加,而且根系Cd含量则大于地上部。当土壤Cd≤50 mg&;#8226;kg^－1时,矿山生态型东南景天根系Cd含量比非矿山生态 型高 ,但当土壤Cd≥100 mg&;#8226;kg^－1,两者之间无显著差异;然而,但在同一Cd处理水平下,矿山生态型东南景天地上部Cd含量总是高于非矿山 生态型。这些结果表明,矿山生态型东南景天有很强的忍耐和吸收土壤Cd的能力,再次证明其为一种Cd超积累植物。     

古老铅锌矿区生态型东南景天对锌耐性及超积累特性的研究

 植物长期生长在重金属污染的生境中,逐渐进化成不同的生态型。通过调查中国东南部古老Pb/zn矿和非矿山生境中的植物种群,发现生长在古老Pb／Zn矿的东南景天(Sedum alfredii Hance)是一种新的Zn超积累植物。在自然和控制条件下,古老Pb／Zn矿生态型比非矿山生态型植株的茎粗、叶片大、植株高。在矿山土壤Zn有效含量为105.5～325.4mg·kg-1时,矿山生态型东南景天植株地上部Zn含量为4134～5000mg·kg-1;当营养液中Zn浓度为1223.6μmol时,其Zn含量高达2％。在相同Zn浓度下,矿山生态型地上部Zn含量比非矿山生态型高30倍左右。两种生态型体内Zn分布也不同,古老铅锌矿山生态型的不同器官中Zn含量以茎>叶片>根系,而非矿山生态型则以根系>茎>叶片。古老铅锌矿山生态型地上部积累的Zn占植株总积累量的90％以上,其中叶片和茎分别占41.66％±5.46％和54.75％±5.87％;非矿山生态型各器官中积累的Zn远远低于古老铅锌矿山生态型,各器官中积累的Zn以茎>根系>叶片。本研究表明,这两种生态型东南景天的发现,为今后探讨植物耐高Zn胁迫和超积累Zn的微进化过程提供了非常有价值的材料,也为Zn污染土壤的植物修复提供了一种很有潜力的候选材料。     

一种新的铅富集植物——富集生态型东南景天

对浙江一古老铅锌矿区的土壤和植物种群进行调查后发现一种新的具有耐铅特性和铅富集能力的植物———景天科景天属东南景天 (SedumalfrediiHance) ,称为铅富集生态型植物。进一步比较和分析了不同浓度硝酸铅处理对富集和非富集生态型东南景天的生长及其对铅的吸收特性的影响。结果表明 ,32 0mgPb/L处理对富集生态型的地上部分生长无显著影响 ,而非富集生态型在 2 0mgPb/L时即出现受害症状。富集和非富集生态型的地上部分铅含量、根系铅含量以及单株铅积累速率均随处理浓度的增加而表现出先升后降的变化趋势。其中富集生态型的地上部分铅含量、根系铅含量以及单株铅积累速率最高可达到 5 14mg/kg、1392 2mg/kg和 8.6 2 μg/plant/d ,分别是非富集生态型的 2 .2 7、2 .6 2和 7.16倍。由于具有生长速度快和高积累铅的能力 ,从植物修复的观点来说 ,东南景天铅富集生态型在铅污染土壤的修复方面具有巨大的潜力。     

镉在水花生叶片中的亚细胞分布及其毒理学

以水花生为试验材料,研究了不同浓度镉(Cd)(0、0.05、0.1、0.2、0.4 mmol·L-1)胁迫下水花生叶中Cd及矿质元素的亚细胞分布、抗氧化能力、叶绿素、可溶性蛋白和植物络合素(PCs)含量的变化.结果表明:随着培养液中Cd浓度的增加,叶片内各亚细胞组分中Cd含量显著增加,但分布不均,主要积累在细胞壁,其次为可溶成分,线粒体和叶绿体中分布最少.Cd处理浓度大于0.2 mmol·L-1时,呈现Cd向水花生叶的可溶成分中分配增加、细胞壁中分配减少的趋势.矿质元素的吸收明显失衡,表现为各亚细胞组分中Ca含量显著增加且主要分布在细胞壁中,细胞壁和可溶成分中P和K含量不断降低,叶绿体中Mg和Fe的含量逐渐减少.叶绿素和可溶性蛋白含量持续下降.谷胱甘肽和抗坏血酸含量先升后降,总抗氧化能力(T-AOC)不断提高,PCs大量累积.水花生对水体中的Cd有一定耐性,且Cd在亚细胞结构中的富集量与其毒性间具有明显的剂量-效应关系;亚细胞中矿质元素的紊乱以及叶绿素和可溶性蛋白含量的显著降低,Cd在亚细胞中的积累对植物体造成了明显的毒害作用;Ca在细胞壁中的累积以及PCs和T-AOC水平的提高说明水花生具有较强的抗性;PCs的合成量与Cd对水花生的毒性变化有一定的相关性,可以作为Cd胁迫的生物标记物.     

高粱属植物对土壤镉吸收及亚细胞的分配

通过盆栽模拟试验研究了3种高粱属植物(甜高粱、高丹草和苏丹草)对土壤cd的富集效应及其亚细胞的分配.结果表明:cd在3种高粱属植物不同部位的富集量随着cd处理浓度的增加而增加,生育期内3种高粱属植物根系中Cd的富集量大于茎鞘和叶片中的富集量;高丹草对Cd的富集量高,甜高梁次之,苏丹草最低;Cd在高粱属植物叶片、茎鞘和根系中各亚细胞组分中的分布相似表现为细胞壁>可溶性部分>细胞核、叶绿体组分>线粒体.     

一种新的铅富集植物--富集生态型东南景天

对浙江一古老铅锌矿区的土壤和植物种群进行调查后发现一种新的具有耐铅特性和铅富集能力的植物--景天科景天属东南景天(Sedum alfredii Hance),称为铅富集生态型植物.进一步比较和分析了不同浓度硝酸铅处理对富集和非富集生态型东南景天的生长及其对铅的吸收特性的影响.结果表明, 320 mg Pb/L处理对富集生态型的地上部分生长无显著影响,而非富集生态型在20 mg Pb/L时即出现受害症状.富集和非富集生态型的地上部分铅含量、根系铅含量以及单株铅积累速率均随处理浓度的增加而表现出先升后降的变化趋势.其中富集生态型的地上部分铅含量、根系铅含量以及单株铅积累速率最高可达到514 mg/kg、13 922 mg/kg和8.62 μg/plant/d,分别是非富集生态型的2.27、2.62和7.16倍.由于具有生长速度快和高积累铅的能力,从植物修复的观点来说,东南景天铅富集生态型在铅污染土壤的修复方面具有巨大的潜力.     

外源钙离子对东南景天生长及锌积累的影响

采用水培试验,研究了外源添加不同浓度钙离子（Ca²⁺）对两种生态型东南景天生物量、根系形态及体内锌、钙、硫含量的影响.结果表明：随着外源Ca²⁺浓度的上升,两种生态型东南景天的干物质量均增加,且超积累生态型地上部增加显著(P<0.05);超积累生态型根长和根表面积增加,而非超积累生态型降低;超积累生态型根、茎、叶锌含量随着外源Ca²⁺浓度的增加而上升,但各处理间差异不显著(P>0.05),非超积累生态型地上部锌含量显著降低(P<0.05).非超积累生态型根、茎、叶钙含量与外源Ca²⁺浓度呈显著正相关(P<0.05),超积累生态型根系硫含量与外源Ca²⁺浓度呈极显著正相关(P<0.01).外源Ca²⁺对超积累生态型东南景天锌吸收及积累有促进作用,而Ca²⁺浓度的升高抑制了非超积累生态型东南景天对锌的吸收.适当增加外源Ca²⁺可促进超积累生态型东南景天生长,改善其锌积累能力.     

Enhanced root-to-shoot translocation of cadmium in the hyperaccumulating ecotype of Sedum alfredii

Sedum alfredii (Crasulaceae) is the only known Cd-hyperaccumulating species that are not in the Brassica family; the mechanism of Cd hyperaccumulation in this plant is, however, little understood. Here, a combination of radioactive techniques, metabolic inhibitors, and fluorescence imaging was used to contrast Cd uptake and translocation between a hyperaccumulating ecotype (HE) and a non-hyperaccumulating ecotype (NHE) of S. alfredii. The K(m) of (109)Cd influx into roots was similar in both ecotypes, while the V(max) was 2-fold higher in the HE. Significant inhibition of Cd uptake by low temperature or metabolic inhibitors was observed in the HE, whereas the effect was less pronounced in the NHE. (109)Cd influx into roots was also significantly decreased by high Ca in both ecotypes. The rate of root-to-shoot translocation of (109)Cd in the HE was >10 times higher when compared with the NHE, and shoots of the HE accumulated dramatically higher (109)Cd concentrations those of the NHE. The addition of the metabolic inhibitor carbonyl cyanide m-chlorophenylhydrazone (CCCP) resulted in a significant reduction in Cd contents in the shoots of the HE, and in the roots of the NHE. Cd was distributed preferentially to the root cylinder of the HE but not the NHE, and there was a 3-5 times higher Cd concentration in xylem sap of the HE in contrast to the NHE. These results illustrate that a greatly enhanced rate of root-to-shoot translocation, possibly as a result of enhanced xylem loading, rather than differences in the rate of root uptake, was the pivotal process expressed in the Cd hyperaccumulator HE S. alfredii.     

Cadmium uptake and xylem loading are active processes in the hyperaccumulator Sedum alfredii

Sedum alfredii is a well known cadmium (Cd) hyperaccumulator native to China; however, the mechanism behind its hyperaccumulation of Cd is not fully understood. Through several hydroponic experiments, characteristics of Cd uptake and translocation were investigated in the hyperaccumulating ecotype (HE) of S. alfredii in comparison with its non-hyperaccumulating ecotype (NHE). The results showed that at Cd level of 10 microM measured Cd uptake in HE was 3-4 times higher than the implied Cd uptake calculated from transpiration rate. Furthermore, inhibition of transpiration rate in the HE has no essential effect on Cd accumulation in shoots of the plants. Low temperature treatment (4 degrees C) significantly inhibited Cd uptake and reduced upward translocation of Cd to shoots for 9 times in HE plants, whereas no such effect was observed in NHE. Cadmium concentration was 3-4-fold higher in xylem sap of HE, as compared with that in external uptake solution, whereas opposite results were obtained for NHE. Cadmium concentration in xylem sap of HE was significantly reduced by the addition of metabolic inhibitors, carbonyl cyanide m-chlorophenylhydrazone (CCCP) and 2,4-dinitrophenol (DNP), in the uptake solutions, whereas no such effect was noted in NHE. These results suggest that Cd uptake and translocation is an active process in plants of HE S. alfredii, symplastic pathway rather than apoplastic bypass contributes greatly to root uptake, xylem loading and translocation of Cd to the shoots of HE, in comparison with the NHE plants.     

伴矿景天植物螯合肽合酶基因的克隆及功能分析

重金属超积累植物由于长期生长在高浓度的重金属环境中,使得经由植物螯合肽（phytochelatins, PCs）解毒途径来应对重金属毒害代价高昂。我们从Zn/Cd超积累植物伴矿景天（Sedum plumbizincicola）中克隆了植物螯合肽合酶（phytochelatin synthase, PCS）基因SepPCS。该基因在裂殖酵母和拟南芥中表达后都具有PCS活性,而且能够互补它们的PCs缺失突变体的Cd敏感表型。SepPCS在伴矿景天中的表达受到高浓度Cd处理的诱导。与其近亲非超积累生态型东南景天（S. alfredii）相比,虽然伴矿景天地上部PCs与Cd的摩尔比远低于东南景天,但是在高浓度Cd处理时PCs含量以及PCs与Cd的摩尔比急剧增加。我们推测在伴矿景天应对Cd毒害的过程中, PCs起到一定的作用,并且在高浓度Cd胁迫时地上部PCs依赖的解毒作用有所加强。     

Cellular sequestration of cadmium in the hyperaccumulator plant species Sedum alfredii

Spatial imaging of cadmium (Cd) in the hyperaccumulator Sedum alfredii was investigated in vivo by laser ablation inductively coupled plasma mass spectrometry and x-ray microfluorescence imaging. Preferential Cd accumulation in the pith and cortex was observed in stems of the Cd hyperaccumulating ecotype (HE), whereas Cd was restricted to the vascular bundles in its contrasting nonhyperaccumulating ecotype. Cd concentrations of up to 15,000 μg g(-1) were measured in the pith cells, which was many fold higher than the concentrations in the stem epidermis and vascular bundles in the HE plants. In the leaves of the HE, Cd was mainly localized to the mesophyll and vascular cells rather than the epidermis. The distribution pattern of Cd in both stems and leaves of the HE was very similar to calcium but not zinc, irrespective of Cd exposure levels. Extended x-ray absorption fine structure spectroscopy analysis showed that Cd in the stems and leaves of the HE was mainly associated with oxygen ligands, and a larger proportion (about 70% in leaves and 47% in stems) of Cd was bound with malic acid, which was the major organic acid in the shoots of the plants. These results indicate that a majority of Cd in HE accumulates in the parenchyma cells, especially in stems, and is likely associated with calcium pathways and bound with organic acid (malate), which is indicative of a critical role of vacuolar sequestration of Cd in the HE S. alfredii.     

Effects of mutual grafting on the cadmium accumulation characteristics of two ecotypes of Solanum photeinocarpum

Abstract

The effects of mutual grafting on the cadmium (Cd) accumulation characteristics of two ecotypes (farmland and mining) of the potential Cd-hyperaccumulator Solanum photeinocarpum were studied through a pot experiment for one month. Four treatments were used in the experiment: ungrafted farmland ecotype (F-CK), ungrafted mining ecotype (M-CK), the farmland ecotype as the scion grafted onto rootstocks of the mining ecotype (F-Scion), and the mining ecotype as the scion grafted onto rootstocks of the farmland ecotype (M-Scion). Mutual grafting increased the rootstock biomass of both S. photeinocarpum ecotypes. However, mutual grafting decreased the scion biomass of F-Scion compared with F-CK and M-CK, and the scion biomass of M-Scion was higher than that of M-CK and lower than that of F-CK. The Cd content in the rootstock of M-Scion increased compared with F-CK, and the Cd content in the rootstock of F-Scion increased compared with M-CK, but mutual grafting decreased the Cd content in scions of both S. photeinocarpum ecotypes. Mutual grafting increased Cd extraction by rootstocks of both S. photeinocarpum ecotypes, but decreased extraction by scions. Therefore, mutual grafting can increase Cd accumulation in S. photeinocarpum rootstocks but not increase Cd accumulation in S. photeinocarpum scions in a short period.