外源甜菜碱对苋菜抗盐性与累积重金属特性的影响

用不同盐分(0.2%、0.4%)、重金属污染土壤进行盆栽实验,研究叶面喷施不同浓度甜菜碱(GB,1、10、50 mmol·L-1)对苋菜抗盐性与重金属累积的影响.结果表明,0.4%重度盐分胁迫下,喷施50 mmol·L-1甜菜碱可大幅提高苋菜茎叶部游离脯氨酸和总游离氨基酸含量;盐分胁迫抑制了苋菜根系对Ca、Mg、K、Fe的吸收,而叶面喷施甜菜碱可显著增强对Ca、Mg、K、Fe的吸收,K/Na、Ca/Na比值随着甜菜碱剂量的增加而逐渐增大.在0.4%盐分下,喷施50 mmol·L-1甜菜碱促使根系K/Na、Ca/Na比值分别增加了40.4%、34.1%,说明盐分越高,外源甜菜碱对苋菜抗盐性更明显.盐分胁迫下,外源甜菜碱明显提高了苋菜根系重金属Cd、Cr、Pb、Cu、Zn的含量.在0.4%盐分与喷施50 mmol·L-1甜菜碱条件下,苋菜根系对各重金属的累积增幅分别为22.1%、127.1%、63.4%、61.1%、56.1%.由根系重金属与盐分离子的相关性分析可知,0.4%盐分处理后,Pb、Cr、Cu、Zn与盐分离子、重金属的吸收变化呈显著正相关(p0.01);而在0.2%盐分处理下,只有Na与Ca、Cd与Cr、Cu与Zn的吸收变化呈显著正相关(p0.01).说明盐分含量较高,外源甜菜碱对苋菜抗盐胁迫效果越明显,对必需元素及其伴随吸收的非必需重金属元素吸收增加也越明显.     

Generation of low-cadmium rice germplasms via knockout of OsLCD using CRISPR/Cas9

The OsLCD gene, which has been implicated in cadmium (Cd) accumulation in rice, might be a useful target for CRISPR/Cas9 editing. However, the effects of OsLCD gene editing on Cd accumulation, plant growth, and yield traits remain unknown. Here, we used CRISPR/Cas9 to generate oslcd single mutants from indica and japonica rice cultivars. We also generated osnramp5 single mutants and oslcd osnramp5 double mutants in the indica background. When grown in Cd-contaminated paddy soils, all oslcd single mutants accumulated less Cd than the wild types (WTs). Consistent with this, oslcd single mutants grown in Cd-contaminated hydroponic culture accumulated significantly less Cd in the shoots as compared to WTs. This decrease in accumulation probably resulted from the reduction of Cd translocation under Cd stress. Oxidative damage also decreased, and plant growth increased in all oslcd single mutant seedlings as compared to WTs in the presence of Cd. Plant growth and most yield traits, as well essential element concentrations in rice seedling shoots, brown rice, and rice straw, were similar between oslcd single mutants and WTs. In the presence of Cd, Cd concentrations in the brown rice and shoots of oslcd osnramp5 double mutants were significantly decreased compared with WTs as well as osnramp single mutants. Our results suggested that OsLCD knockout may reduce Cd accumulation alone or in combination with other knockout mutations in a variety of rice genotypes; unlike OsNramp5 mutations, OsLCD knockout did not reduce essential element contents. Therefore, OsLCD knockout might be used to generate low-Cd rice germplasms.     

水分管理模式与土壤Eh值对水稻Cd迁移与累积的影响

通过水稻盆栽试验,研究了4种水分管理模式[水稻全生育期湿润灌溉(M)、灌浆期湿润和灌浆期后淹水灌溉(M-F)、灌浆期淹水和灌浆后湿润灌溉(F-M)、全生育期淹水灌溉(F)]对土壤Eh值、Cd生物有效性及对水稻Cd迁移与累积的影响.结果表明土壤Eh值从负值变为正值时,土壤交换态Cd含量先升高后降低.与M处理相比,F-M、M-F、F处理显著降低水稻根、茎叶、谷壳、糙米Cd含量,其中M-F和F处理水稻糙米Cd含量分别为0.19 mg·kg~(-1)和0.10 mg·kg~(-1),均低于国家食品污染物限量0.2 mg·kg~(-1)的标准(GB 2762-2012).与M处理相比,F-M、M-F、F处理显著降低水稻地上部Cd累积量和水稻中Cd的转运系数.水稻地上部分Cd累积量、水稻中Cd的转运系数、水稻糙米Cd含量与土壤Eh值均存在正指数关系.4种水分管理模式中,M-F处理的水稻产量最高.虽然威优46被认为是Cd高累积水稻品种,但是在5 mg·kg~(-1)的Cd污染土壤中进行适当的水分管理,威优46糙米Cd含量也能够达到国家标准.因此,为了保证水稻的食品安全和水稻产量,建议采用M-F处理作为水分管理模式,且灌浆期后的土壤Eh值保持在-160~-130 m V范围.     

Piriformospora indica confers cadmium tolerance in Nicotiana tabacum

Piriformospora indica, a root-colonizing endophytic fungus of Sebacinales, promotes plant growth and confers resistance against biotic and abiotic stresses. In order to confirm the influence of P. indica on growth, proline, malondialdehyde (MDA), chlorophyll, and cadmium (Cd) amounts in Nicotiana tabacum under Cd stress, hydroponics, pot and field trials were conducted. The results showed that P. indica can store Cd in plant roots and reduce leaf Cd content, reduce the concentration of MDA, and increase the proline and chlorophyll content and the activities of catalase, peroxidase, and superoxide dismutase under hydroponic Cd stress. RT-PCR analysis showed that the relative expression level of genes Gsh2, TaPCS1, oas1, GPX, and Hsp70 in colonized plants was 4.3, 1.4, 2.9, 1.7, and 6.9 fold higher than in un-colonized plants respectively. Cd exposure significantly reduced un-colonized plants'' agronomic traits compared to P. indica-colonized ones. Our results suggested that P. indica can sequester Cd in roots, so that much less cadmium was transported to leaves, and the increased concentrations of antioxidant enzymes, pigments and proline contents, as well as the higher expression of stress-related phytochelatin biosynthesis genes in P. indica-inoculated plants, may also serve to protect N. tabacum plants against oxidative damage, enhancing Cd tolerance.     

外源锌对水稻各部位镉吸收与累积的拮抗效应

为探明外源锌(Zn)对镉(Cd)污染稻田系统Cd吸收累积的影响,采用盆栽试验,向轻度(Cd含量为0.50 mg·kg~(-1))和中度(Cd含量为1.50 mg·kg~(-1))Cd污染土壤中施用不同浓度的外源Zn,研究Zn对土壤、水稻根表铁膜和水稻各部位中Cd、Zn含量变化的影响,探讨外源Zn对Cd污染稻田系统Cd吸收累积的交互作用.结果表明,在2种Cd污染土壤中,施用外源Zn显著降低了湘晚籼12糙米Cd含量,降低幅度为13.4%～78.4%;在轻度Cd污染土壤中,施用外源Zn显著降低了威优46糙米Cd含量,降低幅度为14.7%～79.8%,而在中度Cd污染土壤中,当外源Zn浓度超过40 mg·kg~(-1)将增加威优46糙米Cd含量;在2种Cd污染土壤下均显著增加2种水稻根表铁膜数量,分别增加14.7%～85.9%、5.8%～61.2%;但对土壤交换态Cd含量无显著影响.在2种Cd污染土壤中,2种水稻各部位Cd/Zn比值与Cd含量正线性相关,且各部位Cd/Zn比值呈下降趋势,2种水稻糙米、茎叶间Cd-Zn拮抗作用显著.轻度Cd污染土壤中,施加外源Zn可以降低湘晚籼12和威优46糙米中的Cd含量;中度Cd污染土壤中,外源Zn仍然可以降低湘晚籼12糙米Cd含量,而当外源Zn施加量高于40 mg·kg~(-1)时将增加威优46糙米的Cd含量.     

Effects of EDTA and plant growth-promoting rhizobacteria on plant growth and heavy metal uptake of hyperaccumulator Sedum alfredii Hance

Phytoremediation is a cost-effective and environment-friendly strategy for decontaminating heavy-metal-contaminated soil. However, the practical use of phytoremediation is constrained by the low biomass of plants and low bioavailability of heavy metals in soil. A pot experiment was conducted to investigate the effects of the metal chelator ethylenediaminetetraacetic acid (EDTA) and EDTA in combination with plant growth-promoting rhizobacteria (Burkholderia sp. D54 or Burkholderia sp. D416) on the growth and metal uptake of the hyperaccumulator Sedum alfredii Hance. According to the results, EDTA application decreased shoot and root biomass by 50% and 43%, respectively. The soil respiration and Cd, Pb, Zn uptake were depressed, while the photosynthetic rate, glutathione and phytochelatin (PC) contents were increased by EDTA application. Interestingly, Burkholderia sp. D54 and Burkholderia sp. D416 inoculation significantly relieved the inhibitory effects of EDTA on plant growth and soil respiration. Compared with the control, EDTA + D416 treatment increased the Cd concentration in shoots and decreased the Pb concentration in shoots and roots, but did not change the Zn concentration in S. alfredii plants. Furthermore, EDTA, EDTA + D54 and EDTA + D416 application increased the cysteine and PC contents in S. alfredii (p < 0.05); among all tested PCs, the most abundant species was PC2, and compared with the control, the PC2 content was increased by 371.0%, 1158.6% and 815.6%, respectively. These results will provide some insights into the practical use of EDTA and PGPR in the phytoremediation of heavy-metal-contaminated soil by S. alfredii.     

碱性废弃物及添加锌肥对污染土壤镉生物有效性的影响及机制

采用盆栽方法,以石灰作对比研究了2种碱性废弃物(纸厂滤泥和赤泥)及添加锌肥对酸性镉污染稻田土壤的修复效应及其相关机制,同时利用大田试验进行了验证.结果表明,盆栽条件下施用石灰、纸厂滤泥和赤泥(2 g·kg^-1)后土壤 pH显著提高,在淹水平衡7 d和水稻移栽后30 d时土壤交换性钙含量显著高于不施的对照,增幅分别为33.1%～76.0%和31.0%～78.3%,同时土壤有效态镉含量显著降低,淹水平衡7 d时降幅为38.4%～45.0%,水稻移栽后30 d和60 d时降幅分别为37.4%～52.9%和33.2%～38.7%,水稻根系和糙米镉含量显著降低,降幅分别为24.0%～48.5%和26.3%～44.7%.等量碱性物质对提高土壤 pH和降低水稻镉累积的效果为石灰＞赤泥＞纸厂滤泥.纸厂滤泥和赤泥添加锌肥(0.2 g·kg^-1)对降低水稻镉累积的效果均比两者单施更明显.大田试验结果趋势基本相同,施入不同量的纸厂滤泥、赤泥及与锌肥配施后两季作物镉累积量显著减少,当季水稻糙米和第二季油菜籽粒镉含量分别较对照降低了27.1%～65.1%和16.4%～41.6%,糙米镉含量基本达到国家粮食卫生标准(GB 2715-2005).因此,对于中轻度酸性镉污染稻田土壤,利用纸厂滤泥、赤泥及与锌肥配合施用是一种可行的修复方法,通过构建与土壤镉污染程度等特征相适应的碱性废弃物用量与锌肥配施比例,可确保稻米质量安全.     

Cadmium accumulation and tolerance of two castor cultivars in relation to antioxidant systems

To investigate the effects of Cd on tolerance and antioxidant activities of castor, two different castor（Ricinus communis L.） cultivars（Zibo No. 5 and Zibo No. 8） were used for a hydroponic experiment（0, 1 and 2 mg/L Cd） and a pot experiment using Cd-contaminated soil（34 mg/kg） with the addition of ethylenedinitrilotetraacetic acid（EDTA）. The results indicated that there were significant differences between the two cultivars with respect to Cd uptake in shoots（113–248 mg/kg for Zibo No. 5 and 130–288 mg/kg Zibo No. 8）, biomass tolerance indexes（64.9%–74.6% for Zibo No. 5 and 80.1%–90.9% for Zibo No. 8） in the hydroponic experiment and survival rates（0% for Zibo No. 5 and 100% for Zibo No. 8）determined by the addition of EDTA in the pot experiment, suggesting that Zibo No. 8 has higher tolerance than Zibo No. 5. Moreover, the castor cultivars have low bioconcentration factors（4.80% for Zibo No. 5 and 5.43% for Zibo No. 8） and low translocation factors（〈1%）.Consequently, Zibo No. 8 can participate in Cd phytostabilization in highly Cd-polluted areas. The results indicated that glutathione（GSH） as a non-enzymatic antioxidant, and antioxidant enzymes including superoxide dismutase（SOD）, catalase（CAT） and guaiacol peroxidase（GPX）, were cultivar- and dose-dependent. The higher tolerance of Zibo No. 8compared with Zibo No. 5 can be attributed to the higher GSH levels in the root and higher GPX activity in the leaf.     

颗粒有机质对水稻镉吸收及转运的影响

采用盆栽试验,通过测定不同颗粒有机质(POM)添加量下土壤、POM中有机碳(POM-C)和Cd(POM-Cd)含量及水稻各部位Cd含量,研究了POM对紫色水稻土水稻镉吸收的影响及机制.结果表明:添加POM提高了土壤有机碳(SOC)、溶解性有机碳(DOC)、POM-C、POM-Cd、POM对Cd的富集系数和土壤有效Cd含量,在POM添加量达2. 5 g·kg~(-1)时可显著提高土壤POM-C和POM-Cd比例.添加POM提高了水稻植株生物量及Cd在水稻植株中的总累积量,但使水稻根Cd含量降低24%～42%,茎叶Cd含量提高9%～30%,籽粒Cd含量在POM添加量为0. 5 g·kg~(-1)和1. 0 g·kg~(-1)时分别降低了17%和36%,添加量为2. 5 g·kg~(-1)时提高了39%.添加POM对Cd在水稻根和茎叶中的分配不显著,但POM添加量较少时可抑制Cd从水稻茎叶向籽粒转运,POM添加达到一定量时,促进Cd从茎叶向籽粒转运,最终提高籽粒中Cd含量.相关分析结果表明,土壤有效Cd是影响茎叶Cd累积的主要因素,POM-Cd总量是影响籽粒中Cd累积的主要因素.因此,添加POM可通过改变土壤SOC、DOC、POM-C、POM-Cd及有效镉含量,影响水稻对Cd的吸收.     

施加盐基离子调控水稻Cd的吸收与累积

盐基离子含量是土壤肥力的重要指标,也是决定土壤Cd生物有效性的重要因素.为探明盐基离子对稻田系统中Cd迁移、转运与累积的影响,本研究通过水稻盆栽试验,按照土壤中盐基离子的物质的量比施加不同浓度梯度KCl、CaCl_2、NaCl和MgCl_2到模拟制备的轻度(0.5 mg·kg~(-1))和中度(1.5 mg·kg~(-1))Cd污染土壤,分析盐基离子对土壤CaCl_2提取态Cd含量和水稻吸收累积Cd的影响.结果表明:施加盐基离子能够增加土壤阳离子交换量(CEC),在轻度和中度Cd污染土壤中,土壤CaCl_2提取态Cd含量分别下降了24.6%～56.1%和17.0%～71.1%,糙米Cd含量分别降低了9.1%～60.5%和7.2%～36.0%.在轻度和中度Cd污染土壤中,糙米Cd含量与土壤CaCl_2提取态Cd含量之间存在线性正相关关系,根、茎、叶Cd含量与Ca含量均呈线性负相关.