龟纹瓢虫对草地贪夜蛾卵和低龄幼虫的捕食作用

为明确龟纹瓢虫Propylaea japonica对草地贪夜蛾Spodoptera frugiperda的控害潜力,在(25±1)℃、L∥D=16 h//8 h、RH (60±5)%的条件下研究了龟纹瓢虫雌、雄成虫和4龄幼虫对草地贪夜蛾卵、1龄和2龄幼虫的捕食能力。结果表明,龟纹瓢虫4龄幼虫和雌、雄成虫对草地贪夜蛾卵、1龄和2龄幼虫的捕食反应均符合HollingⅡ模型。龟纹瓢虫雌、雄成虫和4龄幼虫对草地贪夜蛾卵的日最大捕食量分别为204.6、110.9粒和124.7粒,瞬时攻击率分别为1.125、1.658和1.429,处理时间分别为0.004 9、0.009 0 d和0.008 0 d;对草地贪夜蛾1龄幼虫的日最大捕食量分别为242.4、265.5头和109.4头,瞬时攻击率分别为1.233、1.038和1.411,处理时间分别为0.004 1、0.003 8 d和0.009 1 d;对草地贪夜蛾2龄幼虫的日最大捕食量分别为41.2、40.3头和27.6头,瞬时攻击率分别为1.386、1.226和1.685,处理时间分别为0.024 3、0.024 8 d和0.036 2 d。雌、雄成虫和4龄幼虫对草地贪夜蛾卵和低龄幼虫的搜寻效应均随着猎物密度的增加而降低。龟纹瓢虫对草地贪夜蛾卵和1龄幼虫的捕食量高于2龄幼虫。结果表明龟纹瓢虫对草地贪夜蛾卵和低龄幼虫具有较好的控制作用,为利用龟纹瓢虫控制草地贪夜蛾提供了理论依据。     

利用CRISPR/Cas9敲除葡萄VviPDS1基因的研究

选择葡萄八氢番茄红素脱氢酶(Phytoene desaturase)基因VviPDS1为靶标,利用CRISPR/Cas9系统构建基因敲除载体,瞬时转化葡萄叶片原生质体,检测到不同类型的突变。通过农杆菌介导转化‘无核白’葡萄胚性愈伤组织,筛选获得卡那霉素抗性植株71株。经PCR鉴定,其中53株为阳性植株,阳性率为74.64%。测序结果表明,共有20株在靶点发生不同类型的突变,编辑效率为37.74%;其中9株产生了双等位基因突变。对其进行氨基酸序列预测,在第202位氨基酸之后发生了不同程度的变异。利用CRISPR/Cas9系统敲除VviPDS1获得的突变体植株呈现整体矮化,其叶片出现不同程度白化。表明CRISPR/Cas9系统可以通过细胞中的瞬时或稳定表达进行基因编辑,可以实现在葡萄编辑植株中产生纯合敲除。     

利用CRISPR/Cas9系统敲除葡萄中VviEDR2提高对白粉病的抗性

利用CRISPR/Cas9系统定点编辑葡萄白粉病感病基因VviEDR2（Enhanced disease resistance 2），在VviEDR2的DUF1336结构域设计靶位点VviEDR2-T1，构建CRISPR/Cas9敲除载体，通过农杆菌介导法转化‘无核白’葡萄胚性愈伤组织。对PCR阳性植株进行靶位点扩增测序，结果表明，共有8个转基因植株在靶位点处发生不同类型的双等位基因突变，编辑效率为32%；突变体植株生长势较弱，叶片较小，茎秆丛生、细弱。进一步对突变体植株进行抗病检测，结果表明，接种葡萄白粉菌（Erysiphe necator Schw.）5 d后，突变体植株叶片上白粉菌孢子仅能萌发出少量较短初级菌丝，表皮细胞产生大量明显的H2O2，而野生型叶片中白粉菌萌发出大量初级菌丝、次级菌丝和吸器，无明显H2O2产生。这些结果表明，可以利用CRISPR/Cas9技术编辑葡萄感病基因VviEDR2，提高葡萄白粉菌抗性。     

利用酵母双杂交膜系统筛选与小麦蓝矮植原体免疫膜蛋白互作的异沙叶蝉蛋白

正小麦蓝矮病(Wheat blue dwarf,WBD)是我国西北麦区一种重要植原体病害,在我国西部地区危害严重。该病害由异沙叶蝉(Psammotettix alienus L.)专化性传播,介体传毒成为病害流行的重要中心环节[1]。本实验室前期通过免疫荧光标记研究发现WBD植原体免疫膜蛋白(immunodominant membrane protein, IMP)与介体异沙叶蝉肌动蛋白互作,说明IMP在植原体传播和致病过程中起关键作用。     

Solvent retention capacity and swelling index of glutenin in hard red wheat flour as possible indicators of rheological and baking quality characteristics

To overcome the cost and expense of milling and baking hundreds of samples in wheat breeding programs, cereal chemists have developed various rapid predictive tests for end-use quality assessment. In this study, five small-scale rapid tests, including Solvent Retention Capacity (SRC) and Swelling Index of Glutenin (SIG), were compared for viability and accuracy to determine end-use quality using small quantities of flour. The tests were applied on commercial hard red spring and hard red winter wheat cultivars from two of the three main production regions in South Africa. Rheological and bread-baking characteristics were related to results of the rapid tests. Highly significant cultivar differences were seen for all rheological and baking-related characteristics, and SRC and SIG parameters, with a strong genotype effect. The environment effect was also highly significant across regions, especially for flour protein content (FPC), which affected the rheological and baking-related characteristics differently across the regions. Correlations among SRC, SIG, rheological, and baking quality-related parameters were highly significant, but inconsistent for the two production regions. Only lactic acid SRC and SIG correlated consistently with the rheological and baking quality-related characteristics in both production regions.     

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。     

Exogenous application of melatonin improves plant resistance to virus infection

In this study, melatonin (MEL)-mediated plant resistance to tobacco mosaic virus (TMV) was examined to study local infection in Nicotiana glutinosa and systemic infection in Solanum lycopersicum. Exogenous application of 100 µm MEL increased anti-virus infection activity to 37.4% in virus-infected N. glutinosa plants. The same treatment significantly reduced relative levels of virus RNA analysed by qRT-PCR and virus titres measured by dot-ELISA, and increased the relative expression levels of the PR1 and PR5 genes analysed by qRT-PCR, in virus-infected S. lycopersicum. MEL treatment induced considerable accumulations of salicylic acid (SA) and nitric oxide (NO) but did not significantly affect production of hydrogen peroxide (H₂O₂) in the virus-infected S. lycopersicum plants. Transgenic nahG N. tabacum was used to determine whether MEL-induced TMV resistance was dependent on the SA pathway. The results showed that the relative RNA level of the TMV analysed by qRT-PCR and virus titres analysed by dot-ELISA were not reduced by the MEL treatment in the nahG transgenic N. tabacum seedlings treated twice with 100 µm MEL. The increased relative expression levels of PR1 and PR5 were greatly reduced when cPTIO, an NO scavenger, was included in the MEL treatment. A working model of MEL-mediated plant resistance to TMV is proposed. MEL-mediated plant resistance to viruses provides a new avenue to control plant viral diseases.     

三七NAC基因家族全基因组鉴定与表达分析

为明确三七Panax notoginseng NAC转录因子基因家族的分布、功能和结构,通过生物信息学分析法进行鉴定,对其理化特性、染色体位置和进化特征进行分析,并根据RNA-seq数据分析其家族成员的时空表达模式和受黑斑病菌Alternaria panax诱导后的表达情况。结果显示,三七中共有98个NAC基因家族成员,其编码蛋白质长度介于104～882个氨基酸之间,分子量在11.78～100.20 kD之间,等电点在4.12～9.75之间。这98个NAC基因家族不均匀地分布在三七的12条染色体上,其中1号染色体分布最多（16个）,而11号染色体上分布最少（1个）。三七NAC基因启动子区域存在与光响应、生长素响应、赤霉素响应及茉莉酸甲酯响应等相关的多种顺式作用元件。NAC基因在三七不同组织及根部不同发育时期均有表达,在受到黑斑病菌侵染的叶片中NAC部分基因家族成员显著上调表达。表明NAC基因家族在三七的生长发育和响应黑斑病菌侵染过程中具有重要作用。     

油菜素内酯提高蔬菜作物抗逆性的研究进展

油菜素内酯化合物(Brassinosteroids,BRs)是国际上公认的一类高效、广谱、无毒的新型植物甾醇类生长激素。近年来研究发现,BRs的使用可显著提高植物对生物和非生物逆境的抗性,从而促进植物生长发育,大幅度提高作物产量。本文综述了BRs的类型、应用方式以及在提高蔬菜抵抗干旱、低氧、温度、盐离子、病害、重金属等环境胁迫方面的研究成果,并对未来的研究方向进行了展望。