秸秆还田配施氮肥对稻田土壤活性碳氮动态变化的影响

【目的】土壤微生物量碳氮和水溶性有机碳氮是土壤中最活跃的碳氮组分,是衡量土壤碳氮周转与养分有效性的重要指标。探讨秸秆配施氮肥、氮肥用量及基追比例对稻田土壤微生物量碳氮、水溶性有机碳氮、易氧化有机碳和速效氮的影响,明确秸秆还田条件下水稻生长季不同氮肥用量与基追比的土壤活性碳氮变化特征,为稻麦轮作区秸秆还田的氮肥管理提供理论依据。【方法】2012—2015年在湖北省荆门市田间试验中设置施氮量、秸秆配施氮肥和施氮时期3个大田试验。施氮量：不施氮（N0）,推荐施氮（165 kg·hm ^-2,N165）,习惯施氮（195 kg·hm ^-2,N195）;秸秆配施氮肥：秸秆移除（CK）,秸秆还田（移栽前将上季小麦秸秆全部还田,S）,秸秆还田+习惯施氮量（SN）,秸秆还田+推荐施氮量（SF）,秸秆还田+推荐施氮量+腐解菌剂（SM）;施氮时期：基施﹕拔节期﹕抽穗期氮肥施用比例为7﹕3﹕0（R1）,5﹕3﹕2（R2）,10﹕0﹕0（R3）。【结果】秸秆还田+习惯施氮量（SN）显著提高了水稻拔节期土壤微生物量碳（SMBC）含量,但是其成熟期水溶性有机碳含量（DOC）显著降低。秸秆还田+推荐施氮量（SF）显著提高了水稻拔节期土壤水溶性有机氮含量（DON）。腐解菌剂的施用显著降低了水稻成熟期DON含量,拔节期易氧化有机碳含量（ROC）也显著降低。秸秆还田下增加氮肥用量显著提高了水稻抽穗期和灌浆期土壤速效氮含量（AN）;推荐施氮处理（165 kg N·hm ^-2）的DON和AN含量显著升高;农民习惯施氮处理（195 kg N·hm ^-2）降低了DON和AN含量;增加追施氮肥比例对土壤SMBC和DOC含量无明显影响,但提高了水稻拔节期SMBN和ROC含量。【结论】施氮量及其基追比是影响秸秆还田下稻田土壤活性碳氮含量的主要因素,合理配施氮肥能提高土壤微生物量碳、速效氮及水溶性有机氮等活性碳氮组分含量,增加追肥比例也能提高水稻生育期内土壤活性碳氮含量。     

β-carotene attenuates weaning-induced apoptosis via inhibition of PERK-CHOP and IRE1-JNK/p38 MAPK signalling pathways in piglet jejunum

Weaning may cause oxidative injury, immune response impairment, apoptosis and other injuries in piglets. Oxidative and endoplasmic reticulum stress (ERS) can elicit inflammatory responses, and persistent oxidative and ERS also may lead to apoptotic cascades, which is associated with the pathogenesis of multiple diseases. β-carotene, a natural carotenoid, has potential anti-inflammatory and antioxidant functions. However, the effect of β-carotene on apoptosis in weaned piglets and the detailed molecular mechanism remain unclear. In this study, we found that β-carotene decreased malondialdehyde (MDA) levels and increased the activities of superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px) in piglet serum. β-carotene could inhibit the mRNA levels of caspase-3 significantly, but had no significant inhibitory effect of the mRNA levels of caspase-9 and caspase-12 in the piglet jejunum. In addition, β-carotene decreased the activation of GRP78, CHOP, and JNK/p38 MAPK and the ratio of Bax/Bcl-2. Furthermore, β-carotene had a significant influence on the activation of ERS and apoptosis-related signals in TG-induced IPEC-J2. In the present study, β-carotene pre-treatment attenuated the ratio of Bax/Bcl-2 and prevented TG-induced increases in the level of PERK-CHOP and IRE1-JNK/p38 MAPK pathway activation in a dose-dependent manner. Overall, these findings indicate that β-carotene may protect weaning-induced apoptosis through inhibiting ERS.     

利用CRISPR/Cas9系统编辑日本结缕草ZjSGR基因技术研究

日本结缕草(Zoysia japonica)是一种常见的具有众多优良性状的暖季型草种,秋冬季节在我国北方地区会较早出现叶片脱绿现象,在一定程度上限制了日本结缕草在北方的大面积使用。ZjSGR是从结缕草中分离出来的一种衰老诱导基因,负调控叶片滞绿性状。本研究利用CRISPR/Cas9系统,通过基因枪介导的遗传转化方法实现对日本结缕草ZjSGR基因的编辑,经测序验证获得1株单碱基缺失突变植株。突变植株叶片深绿,在正常生长温度(28~30℃)条件下,和对照相比,叶绿素含量更高(P<0.05)。本研究为进一步阐述ZjSGR基因在日本结缕草滞绿性状调控中的作用机理提供了基础研究材料,也为培育日本结缕草滞绿品种育种工作奠定了基础。     

桓仁水库初级生产力和限制性营养盐研究

2018年4月10日、6月11日、8月14日在桓仁水库的上游(江南九队)、中游(砬砬岗子)和下游(泗河大地)3个站位用500 mL具塞磨口瓶盛装采水器采集的水库水样,进行限制性营养盐原位试验,同时测定水中溶解氧和初级生产力,并检测分析水体理化指标,以查明辽宁桓仁水库氮磷分布、营养盐限制和初级生产力状况,保护桓仁水库生态系统的结构和功能完整性。试验结果显示,营养盐加富试验中,0.5 mg/mL氮+0.3 mg/mL磷的添加组可显著提高水体中的溶解氧水平(P<0.05);上游和中游氮磷比为2∶1、下游氮磷比为4∶1可显著提高水体中的溶解氧水平(P<0.05);初级生产力随季节变化特征明显,原始P/R系数均小于试验组。根据试验结果分析得知,桓仁水库营养盐限制因素受季节变化的影响显著,具有氮、磷双限制等特征;根据初级生产力判断桓仁水库为自养代谢型水体。     

番茄ShARPC5抗白粉病功能分析

【目的】白粉病（powdery mildew）是番茄生产上的重要病害,严重影响番茄的产量。番茄基因组测序工作的完成为抗病基因挖掘提供了重要的信息资源。ARP2/3（actin-related protein 2 and 3）复合体是肌动蛋白微丝骨架动力学的主要调控因子,能够参与包括响应外界胁迫等多种细胞学过程。本研究通过对番茄ARPC5（actin-related protein C5）进行克隆和抗病功能验证,为番茄基因组信息完善、抗病机制解析和分子育种等方面打下基础。【方法】从番茄LA1777（Solanum habrochaites）cDNA中PCR扩增ShARPC5,使用DNAMAN 6.0进行多序列比对;MEGA 6.0构建系统发育树;应用在线工具ProtComp v. 9.0进行亚细胞定位预测。利用实时荧光定量PCR（qRT-PCR）比较接种白粉菌（Oidium neolycopersici,On-Lz）后高感品种Moneymaker（MM）和高抗品种LA1777中番茄ARPC5的表达特征,分析白粉菌侵染与ARPC5表达的相关性。应用病毒诱导的基因沉默（virus-induced gene silencing,VIGS）技术进一步验证该基因在番茄中的抗病功能,观察沉默株和野生型株系接种后表型变化,利用台盼蓝和DAB染色法检测植株产生过敏性坏死和H₂O₂的能力,并检测ShARPC5沉默后一些与植物抗病相关标记基因的表达变化。采用农杆菌浸花法遗传转化拟南芥过表达ShARPC5植株,观察转基因和野生型株系接种后表型变化,并统计单病斑分生孢子数。【结果】从番茄品种LA1777中克隆到ShARPC5,编码132个氨基酸残基,包含一个保守的P16-Arc结构域。与番茄MM-白粉菌亲和互作相比,非亲和互作的番茄品种LA1777在接种白粉菌后,ShARPC5显著上调表达,尤其在接种后18 h。在番茄上沉默ShARPC5能够增加植株对白粉菌On-Lz的敏感性,防卫反应基因PR1b1显著下调表达。组织学观察显示与对照植株相比,ShARPC5沉默植株接种后诱导产生过敏性坏死和活性氧减少。在烟草上瞬时过表达ShARPC5能够诱导产生坏死斑。相反,在拟南芥上过表达ShARPC5能够增加植物的抗病性。【结论】ShARPC5是番茄响应白粉菌侵染的重要基因,可减轻番茄白粉病的发病程度,在番茄抗白粉病机理研究方面具有较大的应用价值,可作为番茄抗白粉病分子育种的一个候选基因。     

植物TIR1/AFBs基因家族研究进展

TIR1/AFBs基因家族是一种存在于细胞核中的生长素受体,属于F-box蛋白基因中的一个小亚族。它们通过与相关生长素相结合活化转录因子来促进基因的表达,从而进行调控,是生长素信号转导过程中的关键部分。为了深入研究生长素信号转导机制,从TIR1/AFBs基因家族的发现与结构,家族成员表达模式的差异及对植物生长发育方面的调节等方面概括介绍了TIR1/AFBs基因家族的分子调控机制,总结了TIR1/AFBs基因的功能。最后探讨了TIR1/AFBs基因的研究方向。     

华山松大小蠹2个甲羟戊酸路径基因的克隆与表达

香叶基二磷酸合酶/法尼基焦磷酸（G/FPPS）和异戊二烯焦磷酸异构酶（IDI）是单萜和倍半萜生物合成途径分支点的关键酶。通过克隆华山松大小蠹DaG/FPPS和DaIDI基因的全长序列，预测基因和编码蛋白的性质、结构和功能，并对序列特征和不同发育时期（幼虫、蛹、初羽化期成虫和扩散期成虫）以及不同组织（头、前中肠、后中肠、后肠和剩余躯体）DaG/FPPS和DaIDI表达的研究，旨在揭示DaG/FPPS和DaIDI在华山松大小蠹信息化学物质合成中的分子调控机制。采用RT-PCR和RACE克隆华山松大小蠹DaG/FPPS和DaIDI基因，DNAStar、Blast、ExpasyProtscale、SignalP 4.1、PSORT和TargetP等多种生物信息学方法对基因全长cDNA序列、基本理化性质、系统进化关系、信号肽和蛋白亚细胞定位等进行分析。用Real-time PCR检测DaG/FPPS和DaIDI在华山松大小蠹不同发育时期和不同组织中的表达。结果表明，克隆获得华山松大小蠹香叶基二磷酸合酶/法尼基焦磷酸DaG/FPPS和异戊二烯焦磷酸异构酶DaIDI全长cDNA序列，分别命名为DaG/FPPS和DaIDI，DaG/FPPS编码的氨基酸序列与山松大小蠹DpF/GPPS相似度最高达95%，氨基酸序列含有一个RALS 四肽基序、7 个异戊烯基转移酶保守区（Ⅰ-Ⅶ）和2个典型的DD**D的天冬氨酸富含区；DaIDI编码的氨基酸序列与山松大小蠹IDI相似度最高，达到92%。Expasy Protscale结果表明DaG/FPPS蛋白疏水性氨基酸明显多于亲水性氨基酸，推测其为疏水性蛋白，而DaIDI蛋白亲水性氨基酸数量大于疏水性氨基酸，故推测DaIDI为亲水性蛋白，有较好的水溶性。信号肽预测结果表明，DaG/FPPS和DaIDI均不含信号肽，TargetP结果表明，DaG/FPPS和DaIDI蛋白含线粒体靶向肽，结合PSORT结果推测DaG/FPPS和DaIDI均定位于线粒体。华山松大小蠹DaG/FPPS和DaIDI在不同发育时期和不同组织中均有表达，其中完全羽化期与扩散期成虫保持一致，该阶段表达量最高，不同组织中前中肠表达量最高。     

Hormonal changes play important roles in the key period of superior and inferior earshoot differentiation in maize

The upper earshoots with higher superiority usually have higher yield potential and higher efficiency. To determine the key period for the asynchronous differentiation of superior and inferior earshoots and how hormones are involved in this process, a two-year experiment was designed using two maize hybrids: Suyu 41 (S41, single-ear hybrid) and AN 101 (A101, double-ear hybrid). The results showed that the lag of lower earshoot differentiation was not only caused by the delay of the differentiation starting time but also related to extension of the duration in spikelet differentiation (stage II) and sexual organ formation stage (stage IV). From 12 days before silking (DBS), the contents of indole-3-acetic acid (IAA), zeatin riboside (ZR)+zeatin (ZT), and gibberellic acid (GA₃) in both upper and lower earshoots of the two hybrids increased dramatically and then decreased quickly. ABA slightly increased in the two hybrids and then decreased slowly in S41, while it was maintained at a high level in A101. At 8 DBS, i.e., the transition period from floret differentiation to sexual organ formation stage, not only the growth of upper-to-lower earshoot difference (ULED), but also the values for ULED of IAA, ZR+ZT and GA₃ were all significantly higher in S41 than in A101. Furthermore, the upper-to-lower hormone ratios IAA_U//AA_L and (ZR+ZT)_U/(ZR+ZT)_L were also much higher in the single-ear hybrid than in the double-ear hybrid, while the GA_3U/GA_3L and ABA_U/ABA_L had no significant differences. In addition, the time course of ULED_hormone/ULED_{earshoot growth rate} also suggested that the hormones work in different ways in earshoot superiority/inferiority formation. The delayed differentiation of lower ear shoots was conclusively related to the later initiation of differentiation and the longer durations of specific differentiation stages. Compared with the regulating roles of IAA and ZR+ZT in the key period (8 DBS) of superiority/inferiority differentiation, GA₃ seems to be affected earlier, while ABA contributes little to this process.