Reduction in cadmium accumulation in japonica rice grains by CRISPR/Cas9-mediated editing of OsNRAMP5

Cadmium (Cd) intake is harmful to human health and Cd contamination in rice grains represents a severe threat to those consuming rice as a staple food. Knockout of Cd transporters is a promising strategy to reduce Cd accumulation in rice grains. OsNRAMP5 is the major transporter for Cd and manganese (Mn) uptake in rice. Nevertheless, it is uncertain whether knockout of OsNRAMP5 is applicable to produce low Cd rice without affecting plant growth and grain yield. In this study, we adopted CRISPR/Cas9-based gene editing technology to knock out OsNRAMP5 in two japonica varieties. We generated three independent transgene-free osnramp5 mutants and investigated the effect of osnramp5 mutations on Cd accumulation and plant growth. Hydroponic experiments showed that plant growth and chlorophyll content were significantly reduced in osnramp5 mutants at low Mn conditions, and this defective growth in the mutants could be fully rescued by supply of high levels of Mn. Cd and Mn accumulation in both roots and shoots was markedly reduced in the mutants compared to that in wild-type plants. In paddy field experiments, although Cd in flag leaves and grains was greatly reduced in osnramp5 mutants, some agronomic traits including plant height, seed setting rate, and grain number per panicle were affected in the mutants, which ultimately caused a mild reduction in grain yield. The reduced plant growth in the mutants can be attributed to a marked decrease in Mn accumulation. Our results reveal that the manipulation of OsNRAMP5 should be treated with caution: When assessing the applicability of osnramp5 mutants, soil pH and soil water content in paddy fields need to be taken into consideration, since they might affect the levels of available Mn in the soil and consequently determine the effect of the mutation on grain yield.     

黄河三角洲台田间渗水池塘的水质变动规律分析

以黄河三角洲地区台田间渗水池塘为研究对象,分析了渗水池塘的水质因子变动规律。结果表明:不同池塘间水质因子差异较大,但均表现出水体盐度较高、氮磷等营养物质含量较低、K+离子含量相对较低、Ca2+离子和Na+离子含量相对较高及K+/Na+显著低于正常海水等特点。结果说明,黄河三角洲地区台田间渗水池塘的水质条件不适合直接进行水产养殖,应对水质调整后才可做养殖之用。     

硒肥与钝化材料组配对土壤Cd钝化及稻米Cd消减效果

为了探讨不同硒肥施用方式联合钝化材料对土壤镉钝化和稻米镉消减的效果,采用盆栽试验的方式,选用亚硒酸钠作为硒肥,钙镁磷肥和硅藻土作为钝化材料,设置基施硒肥+钙镁磷肥+硅藻土和叶面喷施硒肥+钙镁磷肥+硅藻土2种方式,研究其不同用量对镉污染酸性稻田土壤修复与安全利用的影响。结果表明：随着施用量的增加,稻米产量增加,基施硒肥产量略高于叶面喷施硒肥,产量差为 2.115 g/pot,与对照(CK)相比,基施0.28%钙镁磷肥+0.12%硅藻土+0.004‰硒(T3)能够提高1.68倍的稻米产量;随着施用量的增加,pH升高,有效Cd降低,有机质与CEC变化不大;基施硒肥与叶面喷施硒肥处理对土壤pH、有机质与CEC差异不显著,但基施硒肥处理有效Cd含量略低于叶面喷施硒肥处理,T3对土壤Cd的钝化效果最佳;随着基施硒肥用量的增加,稻米Cd含量降低,随着叶面喷施硒肥用量的增加,稻米Cd含量先降低后升高,基施硒肥处理对稻米Cd的消减程度强于叶面喷施硒肥处理,相差 0.021 mg/kg,与对照(CK)相比,T3处理稻米Cd降低0.063 mg/kg。可见,硒对调控稻米镉累积具有重要作用,且基施硒肥强于叶面喷施。综上所述,基施0.28%钙镁磷肥+0.12%硅藻土+0.004‰硒对土壤Cd钝化与稻米Cd消减的效果最佳,值得在镉污染稻田推广应用。     

Fas对镉激活PC12细胞线粒体凋亡通路的影响

旨在探究死亡受体Fas在镉致大鼠肾上腺嗜铬细胞瘤细胞（PC12）凋亡中的作用及其对线粒体通路的调控机制,用10 μmol·L^-1镉处理Fas基因沉默的PC12细胞株12 h,通过Western blot检测BH3相互作用域死亡激动剂（BID）、半胱氨酸蛋白酶-9（caspase-9）、半胱氨酸蛋白酶-3（caspase-3）、多聚二磷酸腺苷核糖聚合酶（PARP）的活化情况,Bcl-2相关X蛋白（Bax）、B细胞淋巴瘤/白血病-2基因（Bcl-2）、凋亡诱导因子（AIF）、核酸内切酶G（Endo G）的表达情况,以及细胞色素C（Cyt C）在细胞内的分布情况,免疫荧光染色检测AIF核转位。结果显示,镉极显著上调tBID/BID比值和Bax/Bcl-2比值,诱导Cyt C从线粒体释放到细胞质,激活caspase-9、caspase-3和PARP,增加AIF和Endo G蛋白表达水平（P<0.01）,并诱导AIF核转位;沉默Fas极显著抑制镉引起的tBID/BID比值和Bax/Bcl-2比值升高,Cyt C从线粒体释放到胞浆,caspase-3、PARP蛋白活化和AIF、Endo G蛋白表达水平极显著升高（P<0.01）,显著抑制镉激活的caspase-9（P<0.05）,并抑制AIF核转位。综上表明,Fas通过调控线粒体通路参与镉致PC12细胞凋亡。     

锰胁迫下鸡眼草的富集特征及生理响应

为探明植物对锰胁迫的耐受性机制,以采自重金属污染区与非污染区的鸡眼草为试验材料,在不同锰浓度胁迫下［0 （对照）、1000、5000、10000、15000、20000 μmol·L^-1］ 开展盆栽试验,比较研究锰胁迫对两种来源鸡眼草表型、生理生化特性及锰积累特征的影响。结果表明：随着锰浓度的升高,1） 两种来源鸡眼草的根干重、芽干重、根冠比均呈降低的趋势;当锰浓度达5000~20000 μmol·L^-1时,与对照相比,污染区、非污染区鸡眼草的芽干重分别下降4.34%~27.71%与16.33%~49.77%,根干重分别下降19.00%~66.06%与27.90%~77.54%,污染区下降幅度均较非污染区的小;2） 两种来源鸡眼草的叶绿素、可溶性糖、脯氨酸含量及超氧化物歧化酶（SOD）、过氧化物酶（POD）活性均呈先升后降的趋势,可溶性蛋白含量逐渐下降,丙二醛（MDA）含量则逐渐升高;3） 两种来源鸡眼草根、茎、叶的锰含量均增加;锰浓度为20000 μmol·L^-1时,污染区和非污染区鸡眼草的叶锰含量分别为对照的16.53和13.41倍。因此,污染区鸡眼草锰耐受能力及富集能力均较非污染区鸡眼草高,较高的抗氧化酶活性是其耐受高锰胁迫的重要生理机制。     

贵州锰矿废渣堆场重金属污染风险评价及草本植物重金属吸收特征

为明确锰矿废渣堆场重金属污染及优势草本植物重金属吸收特征,对贵州省典型锰矿废渣堆场的重金属污染风险进行了评价,并调查研究了锰矿废渣堆场草本植物的类群及重金属吸收特征。共采集优势草本植物18种,隶属11科18属,菊科和禾本科为优势科。对18种草本植物体地上部、地下部及其生长基质中Cr、Ni、Cu、Zn、Cd、Pb和Mn 7种重金属元素进行了测定,结果显示,锰矿废渣堆场重金属污染空间变异较大,Mn污染最严重,其次是Cd,属于Mn、Cd复合型污染。计算了草本植物对7种重金属元素的富集系数、转运系数,并对18种草本植物进行了聚类分析,综合分析了草本植物重金属吸收特征,结果显示,看麦娘对Cd富集系数达22.49,对Cd有较强的富集能力;五节芒、商陆对Cr、Ni具有较强的富集能力和转运能力;风毛菊和夏枯草对锰矿废渣区多种重金属均表现出较强的富集能力和转运能力。研究结果表明,锰矿废渣堆场主要为Mn和Cd污染,矿区本土自然生长的优势草本植物可作为生态修复的首选植物。     

灰铸铁生产技术的一些新进展

对灰铸铁的研究现状和生产中的一些新技术做了评述。重点介绍了高强度薄壁灰铸铁的生产现状以及先进的制造工艺和先进的制造手段在灰铸铁生产中应用所取得的技术成果。     

Heavy Metal Immobilization in Contaminated Soils using Phosphogypsum and Rice Straw Compost

The application of soil amendments to immobilize heavy metals is a promising technology to meet the requirements for environmentally sound and cost‐effective remediation. The present study was carried out to evaluate the result of phosphogypsum (PG) used alone and in combination with compost (CP) at a mix ratio of 1:1 wet weight ratio (PG + CP) at 10 and 20 g dry weight kg⁻¹ dry soil, on heavy metal immobilization in contaminated soil and on canola growth. The results revealed that the Pb, Cd and Zn uptake of canola plants was reduced by the application of PG alone and when it was mixed with CP as compared with untreated soil. At an application rate of 10 g dry weight kg⁻¹ dry soil of (PG + CP) the dry weight of canola plants increased by 66·8% was increased in comparison with its weight in the untreated soil. The addition of PG alone resulted in more pronounced immobilization of heavy metal as compared with PG mixed with CP. Plant growth was improved with CP addition, but heavy metals immobilization was the greatest in PG alone treatments. Results suggest that PG may be useful for the immobilization of heavy metals in contaminated soils. Copyright © 2014 John Wiley & Sons, Ltd.     

Protective Effect of Sulforaphane on Reproductive Function of Male Mice with Cadmium Poisoning

The study was aimed to explore the protective effect of sulforaphane (SFN) on the reproductive function of male mice with cadmium poisoning.40 healthy clean grade male Kunming mice were randomly divided into four groups:control group (H₂O),cadmium chloride group (2.3 mg/kg CdCl₂),sulforaphane group (10 mg/kg SFN),sulforaphane + cadmium chloride group (10 mg/kg SFN+2.3 mg/kg CdCl₂),and continuous administration for 10 d,all mice were executed by dislocated cervical vertebra at 2 d after the last administration,and then the pathologic changes of testicular tissues,organ coefficient of testicle and epididymis,sperm quality and concentration of testosterone were tested.Additionally,the contents of GSH and MDA,and the activities of T-SOD in testis were also detected at the same time. Compared with the control group,pathology damages were observed in cadmium chloride group,organ coefficient of testis and epididymis,sperm quality and levels of testosterone extremely significantly decreased (P<0.01),the activities of T-SOD and GSH content were extremely significantly decreased (P<0.01),and the concentration of MDA was extremely significantly enhanced (P<0.01).Compared with the control group,the activity of T-SOD and concentration of GSH in sulforaphane group were significantly increased (P<0.05),and the concentration of MDA was not significant different between the control group and sulforaphane group (P>0.05).While compared with the cadmium chloride group,the sperm motility rate and sperm total count in sulforaphane and cadmium chloride group were extremely significantly increased (P<0.01),the organ coefficient of testicle and epididymis was increased significantly (P<0.05),the concentration of GSH and activity of T-SOD in testicular tissue were extremely significantly increased (P<0.01),and the concentration of MDA was extremely significantly decreased (P<0.01).The results indicated that sulforaphane had the protection effect on reproduction function of male mice with cadmium poisoning.