Eyespot of Cereals Revisited: ITS phylogeny Reveals New Species Relationships

Four species so far classified in Pseudocercosporella or Ramulispora (hyphomycetes) are associated with eyespot disease symptoms of cereals. Two of these have been linked to teleomorphs that were described in Tapesia. Sequence data derived from the Internal Transcribed Spacer region (ITS1, 5.8S and ITS2) of the rDNA operon showed, however, that the eyespot fungi associated with Tapesia are not congeneric with Ramulispora sorghi, the type of Ramulispora. The genus name Tapesia is now rejected in favour of the conserved name Mollisia, which appears to comprise heterogeneous fungi. Tapesia yallundae is not closely related to the type of Mollisia, M. cinerea, but clusters separately, being more closely allied to species with Cadophora anamorphs. A new holomorph genus, Oculimacula, is therefore proposed for teleomorphs of the eyespot fungi, while the anamorphs are accommodated in Helgardia gen. nov.     

Multiple Selection of Potato Cyst Nematode Globodera pallida Virulence on a Range of Potato Species. I. Serial Selection on Solanum-hybrids

A series of selection experiments on potato cyst nematode Globodera pallida, pathotype Pa1, tested the virulence response of the nematode to a range of resistant potato Solanum genotypes. Alleles conferring virulence against all four Solanum sources used in the study (i.e. Solanum vernei, S. multidissectum, S. sanctae-rosae and S. tuberosum ssp. andigena) were detected. Selection for multiple virulence against a combination of resistant sources resulted in the originally-selected virulence genes being retained or lost. The mechanism, or basis, of potato cyst nematode resistance differs between the Solanum species. The appropriate use of resistance cultivars produced from a range of Solanum-resistant species offers a management tool for controlling potato cyst nematode levels in infested land.     

Evidence that a Leaf-Disk Test Allows Assessment of Isolate-Specific Resistance in Brassica oleracea Crops Against Downy Mildew (Peronospora parasitica)

A rapid resistance/susceptibility test for Peronospora parasitica (downy mildew) was established by inoculating leaf-disks of four Brassica oleracea accessions. Several conditions were tested: disk disinfection or not, agar medium with or without nutrients and with 50 or 100 ppm of benzimidazole. Using disinfected disks placed on agar (no nutrient and benzimidazole at 50 or 100 ppm), the responses of leaf-disks to four isolates were similar to those obtained using the classical cotyledon test, whereas undesired contaminations occurred in all other conditions. The possible effect of the particular leaf used for obtaining the disks was also studied. In each incompatible interaction tested, disks were resistant whatever the leaf used. In compatible interactions, susceptible phenotypes were observed on disks derived from the six lowest leaves, but disks from upper leaves were resistant. The genetic basis of resistance in a F1 hybrid broccoli was assessed, by testing six isolates on an F2 population derived from this hybrid. The cotyledon test only allows inoculation of two isolates per seedling, whereas many isolates can be tested on each plant by using leaf-disks. The segregation of the resistance to each of the six isolates was analysed: two dominant genes (tightly linked) control resistance to all isolates (one to five isolates; the other to only one isolate).     

Identification and Characterisation of Xanthomonas campestris pv. campestris Strains from Tanzania by Pathogenicity Tests, Biolog, rep-PCR and Fatty Acid Methyl Ester Analysis

Black rot, caused by Xanthomonas campestris pv. campestris (Xcc), is a major disease constraint to cabbage production by smallholder farmers in Africa. Variability exists within the pathogen, and yet differentiation of Xcc strains from other closely-related xanthomonads attacking crucifers is often difficult. The Biolog system, fatty acid methyl ester analysis using microbial identification system (MIS), rep-PCR and pathogenicity tests were used to identify and characterise Xcc strains from Tanzania. Great diversity was observed among Xcc strains in their Biolog and rep-PCR profiles. Specific rep-PCR genomic fingerprints were linked to some geographical areas in the country. Most of the Xcc strains were clustered in two groups based on their fatty acid profiles and symptom expression in cabbage although some deviant strains were found. Each of the methods allowed a degree of identification from species, pathovar to the strain level. Biolog and MIS identified all Xcc strains at least to the genus level. Additionally, Biolog identified 47% of Xcc strains to the pathovar and 43% to strain level, whereas MIS identified 43% of the strains to pathovar level. In the absence of a database, the utility of rep-PCR for routine diagnosis of strains was limited, although the procedure was good for delineation of Xcc to the strain level. These findings indicate the existence of Xcc strains in Tanzania that are distinct from those included in Biolog and MIS databases. The limitations noticed warrant continued improvement of databases and inclusion of pathogenicity testing, using universally susceptible cultivars, as an integral part of strain identification.     

基于mtCOI基因序列的我国云南省和缅甸、柬埔寨草地贪夜蛾种群遗传多样性分析

为制定有效的草地贪夜蛾防控措施,分别自缅甸、柬埔寨和我国云南省采集4、2和8个种群共542个草地贪夜蛾样品,基于mtCOI基因序列分析这14个种群的遗传多样性指数、遗传分化系数及基因流。结果表明,缅甸草地贪夜蛾种群的单倍型多样性指数和平均核苷酸差异数分别介于0.273～0.396和4.643～6.727之间,高于我国云南省的草地贪夜蛾种群,分别介于0.047～0.214和0.791～3.636之间;除ZT种群、LS种群和TO种群外,其它11个种群的Fu’s F均为显著正值,表明缅甸、柬埔寨和我国云南省的草地贪夜蛾种群未经历种群扩张事件;在14个种群中,LS种群与其它种群分化较明显,TK种群、MY种群、CP种群、MS种群和KY种群有效迁入个数和有效迁出个数之和较高,分别为699.41、682.50、855.76、684.56和701.31,推测这5个种群在草地贪夜蛾基因交流中起着类似"中转站"的作用。     

基于线粒体COI基因序列的大豆食心虫不同地理种群遗传分化

为揭示大豆食心虫Leguminivora glycinivorella不同地理种群间的遗传分化情况,通过测定19个地理种群214头老熟幼虫的线粒体COI基因序列,利用MEGA 6.0和Dna SP 5.0软件对不同地理种群间序列变异和遗传分化进行分析。结果表明,在214条408 bp的COI基因序列中发现46个变异位点,获得49种单倍型。总群体单倍型多样度为0.8277,固定系数为0.59,基因流为0.17。总群体Tajima’s D检验结果不显著,说明在较近时间内大豆食心虫未经历群体扩张。大豆食心虫不同地理种群间基因交流较少,贵阳种群、都安种群与其它地理种群分化明显,整体的遗传多样性和遗传分化程度较高,地理距离是影响不同地理种群间遗传距离的重要因素。     

Analysis of a Secreted Aspartic Peptidase Disruption Mutant of Glomerella cingulata

Peptidases have been implicated in the pathogenicity of fungi that cause disease in plants. Expression of the secreted aspartic peptidase gene (gcsap), of a Glomerella cingulata isolate pathogenic on apples, is induced during appressorium formation. To determine whether the secreted aspartic peptidase (GcSAP) is essential to pathogenicity, gcsap was disrupted using a vector containing a 637 bp fragment of genomic DNA that encodes the sequence spanning the two active site aspartic acid (Asp) residues. To ensure that the truncated gcsap gene products could not have residual peptidase activity the codons for the active site residues Asp¹¹² and Asp²⁹⁷ were both mutated to histidine residues. Both PCR and Southern analysis confirmed disruption of gcsap. Neither gcsap mRNA nor GcSAP activity was detected in the disruption mutant. Pathogenicity tests on fruit from three apple cultivars showed that GcSAP was not required for pathogenicity. The disruption mutant grew on medium containing protein as the sole source of nitrogen because G. cingulata secretes a previously undetected peptidase(s). A serine peptidase that had a pH optimum between pH 7.0 and 8.0 and a K _m of 0.25 mM for the synthetic substrate succinyl-Ala–Ala–Pro–Phe-p-nitroanilide was identified.     

云南省小麦条锈菌不同地理亚群体的遗传结构分析

为明确云南省小麦条锈菌（Puccinia striiformis f. sp. tritici,Pst）在不同地理环境下的群体遗传结构,通过3种不同地理亚群体划分方式,即以县域（Group C）、区域（Group R）和海拔（Group E）对云南省537个Pst单孢系进行不同层次的群体划分,并利用12对SSR引物对其进行遗传多样性和群体遗传结构分析。结果显示,云南省Pst的遗传多样性水平在Group C的亚群体之间差异最大,且来自滇中及滇东北地区的Pst群体的遗传多样性较高。Group R和Group E的亚群体基因流及遗传分化结果表明,云南省Pst菌源交流频繁、遗传分化较小。系统进化树分析结果显示,滇东北与滇中Pst群体类似,滇东南与滇西Pst群体类似。Pst遗传组分呈现出由东向西、由北向南和自低海拔向高海拔变化的趋势,与云南省自东南向西北逐渐攀升的地形及地势吻合。Mantel检验结果表明地理距离与遗传距离不存在相关性,在8个县域亚群体、2个区域亚群体检测到有性生殖。表明来自滇中和滇东北地区的Pst群体具有更高的遗传多样性,地理隔离可能是滇西地区遗传多样性较低的成因,遗传分化发生在...     

芋疫霉重组聚合酶扩增结合侧流层析试纸条快速检测方法的建立及应用

为建立芋疫霉Phytophthora colocasiae快速准确的分子检测方法,基于Ypt1基因特异序列,设计芋疫霉的特异性引物与探针,建立一种快速、准确、可视化的芋疫霉重组聚合酶扩增结合侧流层析试纸条（recombinase polymerase amplification-lateral flow dipstick,LFD-RPA）检测方法,对该检测方法进行优化,评估其特异性与灵敏度,并对田间疑似样品进行检测。结果表明,优化后的芋疫霉LFD-RPA检测方法最适反应条件为39℃恒温反应30 min。LFD-RPA检测方法能够特异性地检测出芋疫霉,而对其他卵菌近缘种和常见植物病原真菌均未检出,且该检测方法对芋疫霉DNA的检测灵敏度达到1 pg/μL。对田间带病组织检测发现,LFD-RPA检测方法能够快速准确地从田间自然发病植株中检测出芋疫霉。表明本研究所建立的芋疫霉LFD-RPA快速可视化检测方法特异性好、灵敏度高、简单快捷,可用于芋疫病的田间快速诊断。     

Colonization by two isolates of Plasmodiophora brassicae with differing pathogenicity on a clubroot-resistant cultivar of Chinese cabbage (Brassica rapa L. subsp. pekinensis)

A commercial clubroot-resistant F₁ cultivar of Chinese cabbage (Brassica rapa L. subsp. pekinensis), Kukai 70, is resistant to an isolate of populations of Plasmodiophora brassicae from Hagi (HG) city and is susceptible to another from Yamaguchi (YMG) city. The degree and frequency of primary and secondary cycle colonization by the isolates in the root hairs and root tissues of cv. Kukai 70 were compared. Seedlings of cv. Kukai 70 were grown in soils amended with inoculum of either HG or YMG and harvested 10 days after inoculation to observe the primary cycle (number of root-hair infections) and 20, 30, and 40 days after inoculation to observe of the secondary cycle (frequency of infected cells and degree of plasmodial development based on the number of nuclei in infected cells). Although more root hairs were infected in HG than in YMG, fewer cells in root tissues including the cortex and medullary rays were infected in HG than in YMG. In addition, YMG developed plasmodia with many nuclei and formed resting spores, whereas plasmodia remained immature with a small number of nuclei in HG and did not form resting spores even by 40 days after inoculation. These results suggest that suppression of plasmodial development during secondary colonization is associated with resistance mechanisms to HG in cv. Kukai 70. Starch did not accumulate (i.e., development of amyloplasts) in HG-infected cells. This may be involved in the suppression of secondary colonization of P. brassicae in the cultivar.     

茶翅蝽非典型气味受体基因的克隆及其生物信息学特征和组织特异性表达分析

为明确茶翅蝽Halyomorpha halys的非典型气味受体（odorant receptor co-receptor,Orco）基因的生物信息学特征和组织特异性表达谱,采用Trizol法提取茶翅蝽成虫触角的总RNA,利用RTPCR技术克隆其Orco基因,使用生物信息学软件对茶翅蝽Orco蛋白进行跨膜区域预测、结构分析和系统发育分析,并采用实时荧光定量PCR技术测定Orco的组织表达谱。结果表明,克隆获得茶翅蝽Orco基因并命名为Hhal Orco,该基因的开放阅读框全长1 428 bp,编码475个氨基酸。Hhal Orco蛋白的二级结构中,α螺旋占34.11%,无规则卷曲占36.84%,延伸链占29.05%;其分子质量为53.49 kD,等电点为6.16,无信号肽,是非分泌蛋白;有7个跨膜区,是疏水性蛋白,疏水性系数的平均值为0.297;预测的Hhal Orco蛋白为对称亚基构成的同源四聚体。系统进化分析发现,Hhal Orco与同为半翅目的荔蝽Tessaratoma papillosa亲缘关系最近。qPCR检测结果显示,Hhal Orco基因主要在雌雄成虫的触角中表达,在成虫的头、胸、腹、足中仅有微量表达。     

内蒙古亚洲小车蝗种群遗传多样性和遗传分化的ISSR分析

亚洲小车蝗Oedaleus asiaticus Bei-Bienko是我国北方草原和农牧交错区的主要害虫。为评价内蒙古地区亚洲小车蝗种群的遗传多样性和遗传分化,应用ISSR标记方法对内蒙古15个亚洲小车蝗种群遗传多样性及遗传分化进行了分析。结果表明,7条引物扩增出85条ISSR条带,均为多态性条带。多态性比例（P）、Nei''s遗传多样性指数（H）和香农多样性指数（I）分别为82.59%、0.2319和0.3421,表明亚洲小车蝗种群具有较高的遗传多样性。基因流（Nm）和基因分化系数（Gst）分别为1.2298和0.3352,表明亚洲小车蝗不同地理种群具有明显的遗传分化。遗传距离与地理距离呈极显著正相关关系。表明地理距离和地形差异可能是形成亚洲小车蝗种群遗传分化的主要原因。     

甘肃省甘谷县小麦条锈菌群体遗传结构分析

为明确我国小麦条锈菌Puccinia striiformis f. sp. tritici主要越夏地区的群体遗传结构及演变情况,利用SSR荧光检测技术,对甘肃省甘谷县2013—2015年期间连续5个小麦生长季采集的141株小麦条锈菌单孢系基因组DNA进行分子标记分析,对小麦条锈菌季节亚群的遗传多样性进行分析。结果表明,甘谷地区小麦条锈菌的遗传多样性丰富,有效等位基因数为1.71,Shannon信息指数为0.66,2014年秋季(2014A)亚群体的基因型多样性低于其余亚群体。分子变异方差分析结果显示,小麦条锈菌季节亚群体间变异仅占21%,变异主要出现在亚群体内部,表明甘谷地区各季节亚群体间遗传分化水平差异较小,小麦条锈菌群体在一个小范围内基本能维持稳定状态。主坐标分析(PCoA)、遗传分化、基因流以及共享基因型分析均表明2014年秋季(2014A)亚群体的遗传结构与相邻季节亚群体存在一定差异,表明越夏过程对甘谷地区个别年份小麦条锈菌群体周年稳定性造成较大的影响,越冬过程对小麦条锈菌群体的影响相对较小,春季受外来菌源干扰的可能性较低。     

沙葱萤叶甲表皮蛋白基因GdAbd的克隆及对温度胁迫的响应

为揭示沙葱萤叶甲Galeruca daurica表皮蛋白基因GdAbd在其应对温度胁迫中的作用,采用RACE技术克隆表皮蛋白基因GdAbd的cDNA全长序列,并进行生物信息学分析;利用实时荧光定量(qPCR)技术比较GdAbd经不同温度处理1 h及25℃恢复30 min后在沙葱萤叶甲2龄幼虫体内的表达水平。结果表明,GdAbd基因全长708 bp(GenBank登录号:MG874710),开放阅读框为477 bp,编码158个氨基酸;蛋白预测分子量为16.98 kD,等电点为4.26;编码蛋白具有典型的表皮蛋白RR保守结构域,属于RR-2亚族;具有1个跨膜结构和1个信号肽。同源序列比对和系统发育分析表明,GdAbd与马铃薯甲虫Leptinotarsa decemlineata SgAbd-4的同源性最高,氨基酸序列一致性达50.63%。qPCR检测结果表明,与25℃对照相比,GdAbd基因表达水平在-10～5℃低温胁迫时未发生显著变化,而在35℃高温胁迫时发生显著上调;-10、-5和0℃低温胁迫后25℃恢复30 min可诱导GdAbd显著上调表达,但5℃低温和35℃高温胁迫处理与对照差异不显著。说明短时低温胁迫不能显著影响GdAbd表达,但胁迫后回温可诱导其上调表达。     

The Role of the Bark Beetle, Hylastes ater (Coleoptera: Scolytidae), as a Sapstain Fungi Vector to Pinus radiata Seedlings: A Crisis for the New Zealand Forestry Industry?

The role of the bark beetle Hylastes ater in the re-establishment of Pinus radiata forest in New Zealand is discussed. H. ater was found to be a dominant factor in seedlings mortality in the first year following planting. However, seedling mortality is usually relatively low. In contrast, it was found the large numbers of seedlings were sub-lethally damaged by H. ater feeding attempts, particularly in high risk sites. High risk sites were identified as sites that were harvested during March and April (autumn) when the peak flight activity of H. ater occurred, and subsequently planted with P. radiata seedlings the following winter. H. ater was found to vector sapstain fungi to seedlings during feeding attempts, and a strong relationship between the severity of damage and presence of sapstain fungi was identified. The role of H. ater as a vector of these fungi and the potential implications to the New Zealand forest industry are discussed.     

东北地区亚洲玉米螟不同寄主植物种群线粒体基因遗传多样性

为了解不同寄主植物对亚洲玉米螟Ostrinia furnacalis(Guenée)寄主专化性及遗传多样性的影响,以东北地区取食4种寄主植物的亚洲玉米螟种群为研究对象,选取其线粒体COⅠ和COⅡ基因作为分子标记,通过序列比对分析,研究了亚洲玉米螟种群间的遗传多样性、基因流水平及分子变异。结果显示,亚洲玉米螟线粒体基因具有较丰富的遗传多态性;COⅠ和COⅡ基因总寄主植物种群Fu’s Fs检验结果分别为-1.82和-2.04,种群历史呈扩张趋势,基因流Nm分别为6.38和3.24,说明各种群间基因交流水平较高;遗传变异主要来自于种群内部;亚洲玉米螟的单倍型在BI树和NJ树上的分布拓扑结构与寄主植物间无对应关系。研究表明,取食不同寄主植物的亚洲玉米螟种群间的遗传多态性较高,基因交流频繁,且尚未发生遗传分化。     

Description ofPauesia (Pauesia)anatolica (Hymenoptera: Braconidae, Aphidiinae) sp. nov., a parasitoid of the cedar aphidCinara cedri

The first parasitoid species reared from a population of the cedar aphid,Cinara cedri Mimeur, 1935 (Hemiptera: Aphididae), from the peninsula of Anatolia in Turkey is described.Pauesia anatolica (Hymenoptera: Braconidae, Aphidiinae) is closely related to some species of thelaricis group ofPauesia from which it differs in the number of antenomers, the propodeum and features of the female genitalia. Its potential use as a natural enemy in areas where the aphid has been introduced is discussed. http://www.phytoparasitica.org posting Aug. 28, 2005.