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.     

基于新型城镇化背景的耕地利用与保护研究

建国以来，中国城镇化发展经历了六个阶段，呈现出波状起伏、落后于世界水平、依靠政府推动、区域不平衡等特点。城镇化进程中土地利用存在“寅吃卯粮”、农村土地收益流向城镇、土地资源过度开发、耕地健康水平下降、土地政策基本取向难以实现等问题。新型城镇化背景下耕地保护必须有新思维：降低土地消耗强度，提高土地利用效益；建立与农业适度规模经营相适应的农地流转新机制；实施耕地健康养护工程，保障安全食品供给；制定涉地指标省域内跨市有偿调剂和“地随人走”的政策。     

陕南秦巴山地退耕还林工程综合效益评价

为了解退耕还林工程带来的效益,基于2001—2014年陕南地区退耕还林工程的现状,参考国内外有关效益评价指标体系的原则和方法选取指标,建立陕南退耕还林评价指标体系,应用层次分析法确定各项指标权重,利用效益评价模型从生态、经济、社会3个方面对退耕还林的综合效益进行定量评价。结果表明,陕南三市退耕还林效益指数曲线均呈上升趋势,说明退耕还林效益正逐渐、稳定地显现,尤其从2004年开始,退耕还林的效益显现尤为明显,增速加快。汉中、安康、商洛的综合效益较接近且呈现出同步上升的趋势,截至2014年,汉中市退耕效益最高,综合效益指数为0.6630,安康市居中为0.4985,商洛市最低为0.4260。陕南地区退耕还林综合效益逐步显现且仍将保持平稳增长的态势。     

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。     

丘陵山区农田信息采集车底盘爬坡性能仿真

针对丘陵山区精细农作时农田信息数据采集困难的现状,设计了一种新型的丘陵山区农田信息采集车底盘。为了提高该底盘的爬坡性能,运用多体动力学仿真软件RecurDyn中的低速履带模块(Track-LM),建立了该底盘的虚拟样机模型,并对其纵向和横向爬坡性能进行仿真研究,分析了不同坡角和路面对该底盘行驶速度、俯仰角、侧倾角及侧履带压力的影响。此研究可为改善该底盘爬坡性能提供理论依据。     

农田信息管理决策支持系统的设计与实现——基于Silverlight

随着我国农业的发展,农田分布范围不断扩大,产生的农田信息繁多而复杂。基于WebGIS的农业决策支持系统已在一些地区的现代化农业生产中得到了示范应用,但由于其客户端采用Ajax或Flex技术实现,存在多媒体功能不足及平台可移植性较差等问题。为此,借助WebGIS开放性平台,采用微软Silverlight技术,设计开发了基于Silverlight的农田信息管理决策支持系统,实现了地图基本操作、地块信息管理决策及环境信息管理决策等功能。该系统可以辅助用户管理农田信息,并为施肥及生产计划提供决策支持。在黑龙江红星农场的试用结果表明:系统自投入使用后运行正常,并能提供信息管理及适宜辅助决策支持。     

新疆节水灌区农田排水措施分析

随着膜下滴灌技术在新疆的大力推广应用,灌排模式发生了改变,从而导致了新的土壤次生盐渍化问题。在阐述新疆3种排水措施——明沟排水、暗管排水和竖井排灌发展的基础上,分析了目前新疆节水灌区农田排水措施存在的问题,结合南疆某一节水灌区灌排试验,分析了排水措施对土壤盐分及地下水位、矿化度的影响,为当地灌排模式提出合理化建议,且为减轻节水灌区的土壤盐渍化提供参考借鉴。     

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.     

苜蓿地撂荒过程中土壤-微生物-胞外酶及化学计量的变化

紫花苜蓿(Medicago sativa L.)作为优良修复草种在黄土高原广泛种植,然而随着农村人口迁移及土地利用变化,苜蓿人工草地大量弃置的问题已引起广泛关注。为研究苜蓿弃置草地土壤质量变化规律,本研究分析了不同弃置年限(0年、5年和10年)苜蓿草地土壤养分、微生物量、参与土壤碳氮磷(C,N,P)循环的胞外酶活性及其化学计量变化。结果表明：总体上,土壤和微生物C,N含量随弃置年限呈增加趋势,而C,N相关胞外酶活性和化学计量变化幅度较小;土壤和微生物P含量随弃置年限增加而显著降低,但P相关胞外酶活性及化学计量均有不同程度的增加。进一步分析发现：相较于其他土层,表层(0～20 cm)土壤C,N,P含量及其酶活性较高,其变化趋势与总体结果相似;亚表层土壤(20～40 cm)和(80～100 cm)深层土壤C,N,P含量及其酶活性呈先增后减趋势,且各指标较于表层土壤均降低40%～80%。相关分析表明弃置草地土壤由于P匮乏而逐步退化,可通过补施磷肥改善。