基于phy基因的双边界植物表达载体构建

采用同源克隆方法从泡盛曲霉中克隆了1 515 bp 的植酸酶基因,与黑曲霉(A. niger963)phyA2 的核苷酸同源性为95%.以植物表达载体pTTBUG8为基础,构建了由35S启动子调控、具有磷高效利用功能基因(phy)的双边界植物表达载体 pBSP.解决了bar基因及其产物 PAT蛋白的安全性及产权等问题,并为植物养分高效利用基因工程研究奠定了重要基础.     

0601台风“珍珠”暴雨的数值模拟及诊断分析

为了研究台风暴雨过程对局地的影响,利用WRF模式,对0601号强台风“珍珠”引发广东潮州的暴雨过程进行数值模拟,并对模拟的结果进行Q矢量诊断和分析。模拟结果表明,强台风“珍珠”活动期间,低层辐合对潮州的暴雨降水有非常重要的作用,同时配合低层充沛的水汽输送触发了强降水的产生。对WRF模式的输出结果,利用Q矢量进行诊断,发现沿着降水区域移动方向,始终有Q矢量辐合区配合。同时,Q矢量散度的大值区对应降水大值的分布。Q矢量散度在垂直方向的分布,可以用来判断锋生与锋消,对台风的衰减与消亡有一定的预报意义。     

Cucumber mosaic virus infection of chickpea stands: temporal and spatial patterns of spread and yield‐limiting potential

Patterns of spread and yield losses were examined when migrant aphid vectors transmitted Cucumber mosaic virus (CMV) within chickpea (Cicer arietinum) plots. When numbers of chickpea plant infection foci were varied to provide initial infection incidences of 0·3–2%, rate of virus spread and its final incidence increased in proportion to initial virus incidence and pathogen progress curves reflected a polycyclic pattern of spread. Chickpea seed yields decreased by 44–45% when CMV incidence reached 61–74% at final assessment in plots with simulated 1–2% initial incidence. In chickpea plots with or without introduced lupin plant infection foci, cumulative spatial data for diseased plants were assessed using spatial analysis by distance indices (sadie ). When CMV spread within a plot, infection was concentrated in large patch clusters and was consistent with comprehensive localized spread around primary infection foci. When introduced infection foci were absent, there was more diffuse spread with many isolated clusters. In individual plants that developed CMV symptoms at different growth stages, shoot and pod dry weight were reduced by 60–65% and 77–79% (early infection) and 44 and 66% (late infection), respectively. Seed yield losses were 78–80% (early) and 65–67% (late), and reduction in 50‐seed weight was 20–25% regardless of time of infection, so seed number was reduced more by early than late infection. Infection also reduced seed quality as, in addition to smaller seed size, seed coats were discoloured and malformed: the proportions of malformed seeds were 9–11% (early), 3–6% (late) and 0·5% (healthy).     

The invasive weed Ventenata dubia is a host of Barley yellow dwarf virus with implications for an endangered grassland habitat

Ventenata dubia (African wiregrass), a winter annual weed, is a non‐native species invading grasslands, rangelands and pastures throughout the USA. Limited information is available on its suitability as a host to pathogens and insects in its invaded range. The barley/cereal yellow dwarf virus (B/CYDV) complex occurs ubiquitously in Poaceae species. In non‐managed grasslands, BYDV infection influences competitive dynamics between native and invasive grasses and facilitates invasion by non‐native annual weeds. The Palouse prairie of south‐eastern Washington and northern Idaho, USA, is an endangered ecosystem. Surveys of V. dubia in Palouse prairie and neighbouring Conservation Reserve Program (CRP) habitats were conducted to determine whether B/CYDV viral species are present. Laboratory tests examined the suitability of V. dubia to host BYDV‐PAV and serve as an inoculum source. Plant growth and weight parameters were measured to gauge the impact of BYDV‐PAV on V. dubia. Infection of V. dubia in Palouse prairie and CRP habitats with two species of BYDV, PAV and SGV, was detected for the first time. The ability of BYDV‐PAV to infect V. dubia in the laboratory and transmission from infected V. dubia to barley were demonstrated. BYDV‐PAV‐infected V. dubia showed reductions in plant height, number of leaves and tillers per plant, and above‐ground dry weight, suggesting that V. dubia is sensitive to BYDV. Results demonstrate that V. dubia is a host to BYDV and may serve as a virus inoculum source with potential implications for its management, competitive dynamics between invasive and native grasses and future conservation of endangered grasslands.     

New experimental hosts of Barley yellow dwarf virus among wild grasses,with implications for grassland habitats

Barley yellow dwarf virus (BYDV), an economically important virus, infects small grain cereal crops and over 150 other Poaceae species. BYDV infection plays an important role in competition among grasses in non‐managed systems, but many grasses remain unexamined as potential BYDV hosts. This study examined grass species that have not been reported as BYDV hosts but are commonly encountered in non‐managed grasslands throughout the United States and Canada. Laboratory inoculations with BYDV‐PAV using the aphid vector Rhopalosiphum padi were performed to examine the ability of 13 grass species and barley to be infected with the virus; eight of the grass species were not documented previously as virus hosts. Serological and molecular assays were used to confirm BYDV‐PAV infection. Plant height, number of leaves, number of tillers and weight were recorded to evaluate susceptibility or sensitivity to BYDV. Infection with BYDV was experimentally achieved for the first time on Achnatherum lettermanii, Achnatherum occidentale, Achnatherum thurberianum, Danthonia intermedia, Poa fendleriana, Sporobolus airoides and Sporobolus cryptandrus, but not on Alopecurus pratensis and Elymus wawawaiensis. Infection was confirmed in Bromus inermis, Elymus elymoides, Poa bulbosa, Poa secunda and Hordeum vulgare, which served as controls. BYDV infection caused reductions in plant height on P. bulbosa and P. fendleriana. BYDV‐infected P. secunda had more leaves per plant compared to healthy plants of the same species. BYDV‐infected A. lettermanii exhibited reduced dry weight in both below‐ground and above‐ground tissue. These findings have implications for the management and conservation of grassland habitats.     

拟南芥rd29A启动子的克隆及植物表达载体构建

以拟南芥(Arabidopsis thaliana)基因组DNA为材料,根据Gen Bank中拟南芥rd29A启动子序列设计引物,采用PCR克隆一段长度为951 bp的rd29A启动子序列。采用Plant CARE启动子在线预测工具分析表明,rd29A启动子序列具有TATA-box、CAAT-box基本的顺式作用元件和1个脱落酸应答元件(ABRE)和2个脱水应答调控元件(DRE)。将rd29A启动子序列亚克隆到pcambia3301载体的多克隆位点,构建了由rd29A启动子驱动的植物表达载体。     

RNAi介导的百合抗CMV和LMoV的载体构建

为获得高效的RNAi(RNA interference)表达载体,培育百合抗病毒新种质,选取感病百合叶片为试验材料,同源序列比对后选取CMV 2b基因271 bp和LMoV cp基因428 bp作为靶基因片段。首先采用RT-PCR方法对感病植株进行病毒检测,确定其带有CMV和LMoV病毒;之后提取感病植株叶片总RNA,反转录得到cDNA,设计带有不同酶切位点的引物,以反转录得到的cDNA为模板,PCR扩增获得目的片段,产物回收纯化后,转化大肠杆菌,重组克隆。将各自的反向片段、正向片段顺次连接到载体pFGC5941上,经多次双酶切、连接、转化后,采用冻融法将构建好的载体转入农杆菌EHA105并进行重组鉴定。酶切结果表明已成功构建了分别抗CMV、LMoV的RNAi植物表达载体pFGC5941-C2和pFGC5941-L2,PCR扩增结果表明表达载体已成功转入农杆菌中。所构建的载体及获得的工程菌与预期的完全一致,获得含载体的农杆菌可以直接应用于百合转基因抗病毒育种工程中。     

Detection of the white spot syndrome virus in zooplankton samples collected off the coast of Sonora,Mexico

The aim of this study was to determine the presence of the white spot syndrome virus (WSSV) in zooplankton organisms collected from an important shrimp‐culture area of the Gulf of California. Environmental water parameters (temperature, salinity, dissolved oxygen, and pH) were monitored, and zooplankton was collected. Samples were sectioned according to taxonomic groups (from Phylum to Family) for the detection of WSSV via PCR. A total of 52 zooplankton taxa were identified, of which crustaceans were the most abundant (82%). From the 228 WSSV diagnoses performed, 35% were positive. Moreover, from 32 taxa recorded at least one positive result was observed during the study, and only 13 were negative in all tests. The highest prevalence was observed in three taxa of copepods, brachyurous and bivalves. However, considering prevalence and frequency of occurrence, it was determined that up to 12 taxa could be considered as high‐risk vectors. Finally, a significant correlation was found between the number of diagnoses per station with biovolume (rs = 0.817) and taxa richness (rs = 0.995). The results suggest that zooplankton diversity and abundance are associated with virus dispersion; moreover, these results demonstrated that the dispersion capacity of the virus through different taxa has been probably underestimated in recent years.     

种子带毒率及有翅蚜降落量对SMV流行的影响

大豆田主要迁飞降落的蚜虫有大豆蚜、棉蚜、玉米蚜、桃蚜、禾谷缢管蚜、豆蚜等,而大豆蚜的数量占绝对优势。当有翅蚜迁飞降落量大,同时田间病株率高时,SMV日侵染率高,最高达49.57%。田间有翅蚜发生和迁飞降落高峰一般在7月份,此时的迁飞降落蚜量和SMV的种子带毒率决定病害流行的程度。0.5%种子带毒率显著推迟SMV流行的时期。田间有翅蚜发生和飞翔高峰期推迟到8月份,0.2%和0.1%种子带毒率可控制SMV的流行为害。