miR-18a-5p在鸡不同生长时期的表达及其生物信息学分析与靶基因预测

为探究miR-18a-5p在鸡不同生长时期组织表达变化规律及其生物信息学特点,本研究以苏禽3号鸡为试验动物,利用实时荧光定量PCR技术检测不同生长时期鸡miR-18a-5p的组织表达变化。通过文献和miRBase检索脊椎动物的miR-18a-5p序列,利用Ensembl数据库确定miR-18a-5p在基因组中的位置,根据成熟序列构建系统进化树;使用miRmap、microT、miRanda和TargetScan网站预测miR-18a-5p靶基因,并进行GO和KEGG分析。组织表达分析结果显示,miR-18a-5p序列在鸡心脏、脾脏、肾脏和下丘脑中的表达量显著高于除大脑以外的其他组织(P<0.05)。与3日龄雏鸡相比,90日龄鸡心脏、脾脏、肾脏、腿肌和下丘脑中miR-18a-5p的表达量均显著上升(P<0.05)。在50种脊椎动物中共发现52条miR-18a-5p序列,几乎所有物种都只有1条成熟序列;基因定位分析发现,鸡miR-18a-5p位于1号染色体上的基因间隔区。多序列比对分析表明,不同物种miR-18a-5p的成熟序列同源性较高,物种间较为保守。系统进化树分析发现,鸡miR-18a-5p与原鸽、斑胸草雀等其他鸟类聚为一类,这表明miR-18a-5p进化过程中是保守的。靶基因预测和功能分析发现,miR-18a-5p共有121个靶基因。GO分析结果显示,靶基因主要富集到蛋白质泛素化、细胞周期阻滞、白细胞介素-6产生的负调控等功能。KEGG 通路分析表明,靶基因主要富集到细胞周期及Wnt和FoxO信号通路等。多个与肌肉生长及细胞增殖等相关的基因富集到相关通路之上。综上,鸡miR-18a-5p是组织广泛表达的miRNA,其可能通过Wnt及FoxO信号通路调控肌肉生长及细胞增殖分化。本研究为miR-18a-5p功能及调控机制的深入研究提供参考依据。     

5‐Aminolevulinic Acid Activates Antioxidative Defence System and Seedling Growth in Brassica napus L. under Water‐Deficit Stress

The present study assesses the effects of 5‐aminolevulinic acid (ALA, 0, 0.1, 1 and 10 mg l^?1) on the growth of oilseed rape (Brassica napus L. cv. ZS758) seedlings under water‐deficit stress induced by polyethylene glycol (PEG 6000, 0 and ?0.3 MPa). Water‐deficit stress imposed negative effects on seedling growth by reducing shoot biomass, cotyledon water potential, chlorophyll content and non‐enzymatic antioxidants (glutathione and ascorbic acid) levels. On the other hand, water‐deficit stress enhanced the malondialdehyde (MDA) content, reactive oxygen species (ROS) production, enzymatic antioxidants activities, reduced/oxidized glutathione ratio (GSH/GSSG) and reduced/oxidized ascorbic acid (ASA/DHA) ratio in seedlings. Application of ALA at lower dosages (0.1 and 1 mg l^?1) improved shoot weight and chlorophyll contents, and decreased MDA in rape seedlings, whereas moderately higher dosage of ALA (10 mg l^?1) hampered the growth. The study also indicated that 1 mg l^?1 ALA improved chlorophyll content, but reduced MDA content and ROS production significantly under water‐deficit stress. Lower dosages of ALA (0.1 and 1 mg l^?1) also enhanced GSH/GSSG and ASA/DHA as compared to the seedlings under water‐deficit stress. The antioxidant enzymes (ascorbate peroxidase, peroxidase, catalase, glutathione reductase and superoxide dismutase) enhanced their activities remarkably with 1 mg l^?1 ALA treatment under water‐deficit stress. It was also revealed that 1 mg l^?1 ALA treatment alone induced the expression of APX, CAT and GR substantially and under water‐deficit stress conditions ALA treatment could induce the expression of POD, CAT and GR to a certain degree. These results indicated that 0.1–1 mg l^?1 ALA could enhance the water‐deficit stress tolerance of oilseed seedlings through improving the biomass accumulation, maintaining a relative high ratio of GSH/GSSG and ASA/DHA, enhancing the activities of the specific antioxidant enzymes and inducing the expression of the specific antioxidant enzyme genes.     

绿皮苦瓜新品种冠春5号的选育

冠春5号是以高代自交系327作母本,以自交系186作父本配制而成的苦瓜一代杂种。植株生长势旺盛,早熟,从播种至始收82 d(天)。主侧蔓均可结果,主蔓第1雌花节位为第7~8节。瓜长棒形,瓜长25~35 cm,横径5~7 cm,肉厚0.9~1.0 cm,单瓜质量420 g左右。果表皮浅绿色,点状突起与短条状突起相间瘤状物,尾部钝,果肉脆,苦味淡。早春栽培一般每667 m~2产量约3 000 kg,田间对白粉病和疫病的抗性优于对照新农村油绿苦瓜。适宜四川、重庆等地种植。     

生防菌+恶霉灵最优组合的筛选及对苦瓜枯萎病的防治效果研究

苦瓜枯萎病是由尖孢镰孢菌苦瓜专化型（Fusarium oxysporum f. sp. momordicae）侵染引起的一种土传病害，目前市场上缺乏防治效果较好的农药或生防菌剂。为了更好地防治苦瓜枯萎病，通过盆栽试验筛选出绿针假单胞菌（Pseudomonas chlororaphis）G5 与恶霉灵复配施用的最优组合，并在苦瓜枯萎病发病样地对盆栽试验筛选出的最优组合进行防效验证。试验结果表明：菌药组合B+0.75Hym（1×10⁹ cfu · mL^-1 G5 菌悬液+187.5 mg · L-¹ 恶霉灵）处理对盆栽苦瓜枯萎病的防病效果为81.60%，明显高于其他处理；同时该组合对苦瓜幼苗的促生作用最为明显。接种苦瓜枯萎病菌后，菌药组合B+0.75Hym的苦瓜叶片中PAL、PPO、POD 防御酶活性较高。田间防效验证试验中苦瓜苗移栽30、60 d 后菌药组合B+0.75Hym 对苦瓜枯萎病的防治效果较高，分别达到了52.59%、32.13%，且苦瓜产量也高于其他处理。表明盆栽试验筛选出的菌药最优组合B+0.75Hym 不仅能够降低农药的使用量，而且能够有效防治苦瓜枯萎病，提高苦瓜产量，有一定的生产实用价值。     

质核互作型不育系阜CMS5A大豆的选育及应用

阐述了质核互作型不育系阜CMS5A大豆的选育经过,总结了其特征特性,并分析了其应用成效,以供参考。     

小麦转录因子TaMYB5 like转录自激活能力及其参与生理型花粉败育过程表达模式研究

化学杂交剂SQ-1诱导的小麦花粉败育是一个复杂的生理代谢过程，TaMYB5-like被发现参与此过程。为了解TaMYB5-like基因在SQ-1诱导小麦生理型花粉败育过程中的作用，以SQ-1诱导的小麦生理型雄性不育系PHYMS-1376和对照可育系CK-1376四分体、单核早期、单核晚期、二核期和三核期花药为试验材料，克隆TaMYB5-like基因，并对其进行序列结构、生物信息学、表达模式、亚细胞定位及转录自激活效应分析。结果发现，TaMYB5-like基因序列长5 207 bp，其cDNA长为1 133 bp，其中CDS序列长819 bp，编码272个氨基酸，含有2个SANT结构域，分子量为29.36 kDa，无信号肽，无跨膜结构，亚细胞定位在细胞核内；TaMYB5-like基因启动子涉及光响应、逆境胁迫响应、茉莉酸甲酯响应等顺式作用元件。自激活效应发现，TaMYB5-like蛋白的自激活区段位于N端，合适浓度的AbA和3-AT可以抑制BD-TaMYB5-like和BD-TaMYB5-like-N-SANT融合蛋白的自激活性；随着蛋白质氨基酸序列的截断，自激活性呈降低趋势。与CK-1376相比，PHYMS-1376四分体和单核早期花药中TaMYB5-like表达量差异不显著；花药发育到单核晚期、二核期和三核期时，TaMYB5-like表达量显著增加；CK-1376和PHYMS-1376三核期时花药内TaMYB5-like表达量均最高。     

A calibration method of detecting soil water content based on the information-sharing in wireless sensor network

In agricultural production, there is contradiction between the cost and accuracy of detection during the course of acquiring soil water content (θ) online. This conflict is one of the core issues of automatic water-saving irrigation technology in agriculture. At the same time, capacitive soil moisture sensor (CSMS) has received considerable attention, for it can acquire θ with low cost and high precision, and meet the application requirements of wireless sensor network (WSN). But CSMS is vulnerable to the soil temperature (T_s) and salinity (S_s) in the measurement process. Therefore, this study took EC-5 sensor for example to establish water detection calibration models of soil temperature and salinity for single sensor, using Least Squares Support Vector Machines on MatLAB (LS-SVMlab) as the tool. On this basis, we explored the spatial variability of T_s and S_s, and then a method, which could be used to calibrate the output signals of sensors in multi-point network, was proposed based on the information-sharing (T_s or S_s) technology of WSN. Through laboratory experiment, we effectively reduced the impact of soil temperature and salinity on the single sensor. In example analysis, we investigated the detection precision and costs under different information-sharing radiuses (r). And the results indicated that the method we proposed based on the information-sharing technology of WSN could successfully calibrate the influence of soil temperature and salinity on sensors in multi-point network, and it was an efficacious approach to determine the balance between the calibration accuracy of moisture sensor and the investment of agricultural production. For example, while the calibration precision of soil temperature and salinity is respectively 1%, the costs can be reduced by 30%.     

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。     

不同极性色谱柱在滇红香气成分分析中的对比研究

采用顶空固相微萃取(HS-SPME)对滇红的香气成分进行富集,分别使用两种不同极性的色谱柱进行色谱分离,气质联用(GC-MS)检测分析,比较了非极性色谱柱HP-5MS和极性色谱柱CP-Wax对红茶香气成分分离效果的差异。结果表明,两种不同极性色谱柱对红茶香气成分的分离效果均表现良好,但分析结果存在一定的差异。因此在对红茶中不同的香气成分进行研究分析时,应根据分析目标的不同选择合适的色谱柱。