Genetic diversity of the pine pathogen Lecanosticta acicola in Slovenia and Croatia

Brown spot needle blight (BSNB), a disease of pine trees caused by the fungus Lecanosticta acicola, has been known in Slovenia since 2008 and in Croatia since 1975. Recent outbreaks in Slovenia prompted this study to compare L. acicola populations in these two neighbouring European countries. Sixty-nine isolates collected from three pine species (Pinus mugo, P. halepensis and P. nigra) were used to determine the phylogenetic relationships, genetic structure, and reproductive strategy of the pathogen. EF1-α sequences showed that Slovenian and Croatian isolates share a common ancestry with individuals from central and northern Europe. Population structure analysis revealed four distinct population clusters of L. acicola in these two countries, generally corresponding to their respective geographic location and host. An unequal ratio of mating types and a low overall genetic diversity in the population indicated a strong influence of asexual reproduction. Although some of the oldest recorded European occurrences of BSNB are from Croatia, this study provided no evidence that the population studied in Croatia was the source of the sampled outbreaks in Slovenia. Recent outbreaks of L. acicola in Slovenia are most likely due to introductions from other, yet to be identified, sources.     

玉米大斑病菌和小斑病菌交配型多重PCR 检测方法的建立与应用

【目的】由大斑突脐蠕孢（Exserohilum turcicum）和玉蜀黍平脐蠕孢（Bipolaris maydis）引起的玉米大斑病和小斑病是玉米生产上重要的叶部真菌病害,本研究旨在建立玉米大斑病菌和小斑病菌交配型多重PCR检测方法,为玉米大斑病菌和小斑病菌的交配型田间分布和有性生殖研究提供技术方法。【方法】根据GenBank中已登录的玉米大斑病菌（登录号MAT1-1：GU997138和MAT1-2：GU997137）和小斑病菌（登录号MAT1-1：X68399和MAT1-2：X68398）交配型基因序列,利用Primer Premier 5.0软件设计2种病原菌交配型多重PCR检测特异性引物,采用单因素法对引物的退火温度以及扩增程序中延伸时间和循环数等重要参数进行优化,建立玉米大斑病菌和小斑病菌交配型多重PCR检测方法,并对2种病原菌的交配型多重PCR检测灵敏度和特异性进行检验。同时,对田间采集的129株玉米大斑病菌和194株玉米小斑病菌单孢菌株的交配型进行多重PCR检测,以明确建立的交配型多重PCR检测方法的适用性。【结果】建立的多重PCR方法和设计的交配型特异引物StMAT01-2F/R、StMAT02-3F/R、ChMAT01-3F/R和ChMAT02-2F/R可分别扩增出MAT1-1、MAT1-2型菌株大小为816、132 bp（大病斑菌）与490、136 bp（小病斑菌）的特异性目的条带。25 μL多重PCR扩增体系：2×Multiplex PCR Mix 12.5 μL,引物各10 pmol,DNA模板100 ng,退火温度为57.2℃（大病斑菌）和55.0℃（小斑病菌）,35个循环。该多重PCR对玉米大斑病菌MAT1-1、MAT1-2型单孢菌株的检测灵敏度分别为0.1、0.01 ng基因组DNA,而对玉米小斑病菌MAT1-1、MAT1-2交配型的检测灵敏度均为0.1 ng基因组DNA。该交配型多重PCR检测方法对玉米大斑病菌和小斑病菌特异性很强,能够很好地区分玉米大斑病菌和小斑病菌相应的近缘种和14株其他真菌菌株。不同地理来源的玉米大斑病菌和小斑病菌交配型检测结果表明,该多重PCR能够准确地检出129株玉米大斑病菌和194株玉米小斑病菌的交配型,且检测结果与随机抽取的菌株杂交验证结果完全吻合。【结论】构建的玉米大斑病菌和小斑病菌交配型多重PCR检测方法灵敏度高、特异性好、操作简便,能够准确、快速地检测出玉米大斑病菌和小斑病菌的交配型,为玉米大斑病菌和小斑病菌的交配型田间分布和监测及有性生殖研究提供了可靠的技术方法。     

3种龙胆科植物的资源分配对策

选择青藏高原东缘同一生境类型下具有不同交配系统特点和不同传粉者丰度的3种龙胆科植物湿生扁蕾(Gentianopsis paludosa)(自花授粉)、祁连獐牙菜(Swertia przewalskii)(高昆虫访问异花授粉)和线叶龙胆(Gentiana farreri)(低昆虫访问异花授粉)为对象,研究其资源分配特征。结果表明,1)3种植物的花粉胚珠比例(P/O)存在显著差异(P0.05),湿生扁蕾的P/O显著低于另外两种异花授粉植物的(P0.05),祁连獐牙菜的P/O显著低于线叶龙胆的(P0.05);2)3种植物的繁殖分配比例存在差异,湿生扁蕾的繁殖分配比例显著高于另外两种异花授粉植物的(P0.05),线叶龙胆的繁殖分配比例显著高于祁连獐牙菜的(P0.05);3)在繁殖分配比例与个体大小关系上,湿生扁蕾与其个体大小不存在显著相关关系(P0.05),而另外两种异花授粉植物则与其各自个体大小之间存在显著负相关关系(P0.05)。研究表明,交配系统类型和传粉者丰度与植物的资源分配模式有关,尽管3种植物的繁殖绝对投入量都与其个体大小显著正相关(P0.05),但自花授粉植物相对于异花授粉植物具有较高的繁殖分配比例,并且这种较高的繁殖分配比例不受个体大小影响,这种高繁殖投入的资源分配模式可能与自花授粉植物具有繁殖保障,从而降低其繁殖代价有关。     

金乌贼雄性规格和社群数量对求偶与交配行为的影响

为探讨不同规格雄性金乌贼(Sepia esculenta)在求偶交配过程中的竞争及优势等级,采用实验生态学方法,在室内可控条件下分别设置1L1S组、1L2S组和1L4S组(L和S分别表示大规格雄性和小规格雄性,数字表示实验中雄性金乌贼数量),连续摄像观察和记录金乌贼的繁殖行为。结果显示:(1)繁殖过程中金乌贼具有明显的领域行为和护卫伴游行为。(2)随着群体中小规格雄性比例的增加,处于优势地位的大规格雄性个体的优势等级发生变化,主要表现为大规格雄性护卫伴游时间、成功交配次数逐渐减少,而小规格雄性护卫伴游时间、成功交配次数逐渐增多。(3)当群体中小规格雄性较少时,大规格雄性常常主动攻击小规格雄性,而当小规格雄性数量远大于大规格雄性时,小规格雄性主动向其他雄性发起攻击,以争取护卫权和交配权。研究表明,雄性金乌贼的规格和数量对于求偶竞争以及交配行为具有显著影响。随着繁殖群体中小规格雄性增多,大规格雄性成功交配次数逐渐降低。1L4S组成功交配次数稍高于1L2S组,显著高于1L1S组(P0.05)。人工苗种繁育过程中选择合适规格的雄性亲体和合理的雌雄比例,对提高金乌贼繁殖效率具有重要意义。     

花色多态植物紫薇花部综合特征与繁育系统

[目的]紫薇具有红色、紫色和白色等显著不同的花色以及形态、大小和颜色明显不同的2种雄蕊,使其成为花色多态型和异型雄蕊研究的良好材料.对紫薇花部综合特征和繁育系统进行系统研究,探究紫薇不同花色植株的繁殖策略以及2种雄蕊在传粉过程中的功能作用.[方法]通过野外定点观测,采用传粉昆虫调查、人工授粉以及人工控制套袋试验等方法对紫薇3种花色植株的花部形态特征、开花进程、传粉式样以及花朵的功能形态特征进行检测.[结果]1)3种花色植株的花期和单花开放进程基本相同(单花花期为4～5天).2)3种花色植株的花在形态特征上存在差异.3)3种花色植株的短雄蕊数表现为白色花植株＞紫色花植株＞红色花植株,花色间存在显著差异(P＜0.05);红色花植株的单花花粉量和P/O值显著低于其他2种花色植株(P＜0.05).4)3种花色植株的花粉活性和柱头可授性动态变化趋势相似;花粉寿命与柱头可授期有部分重叠,长雄蕊花粉活性最高时期早于柱头最佳可授期,短雄蕊的与柱头最佳可授期同步;白色和紫色花植株的柱头最佳可授期比红色花植株的长(约2h).5)2种雄蕊在形态、颜色、花粉量、人工控制试验条件下的结实率和结籽数以及传粉昆虫访花行为等方面都存在显著差异,说明2种雄蕊在传粉过程中存在功能分化,即黄色短雄蕊主要起招引昆虫作用,暗黄色长雄蕊主要起传粉作用;但2种雄蕊在花粉活性、花粉组织化学成分以及人工控制授粉后的结实率等方面均无显著差异,表明二者在生理上尚未分化;去除雄蕊试验表明,雄蕊的排列方式是影响2种雄蕊功能分化程度的重要因素.6)P/O值、套袋试验以及传粉昆虫观察结果表明,紫薇不存在无融合生殖、主动自交和自交不亲和现象,其交配方式是以异交为主、自交亲和的兼性异交繁育系统.7)传粉昆虫对红色花植株有明显偏好,访问该花色植株的频率显著高于其他2种花色植株(P＜0.01);人工授粉后,白色和紫色花植株的结实率显著提高.[结论]在自然条件下,紫薇的白色和紫色花植株存在一定的花粉限制.此时,增加对雄性功能的投入、延长柱头最佳可授期是其维持繁殖成效的适应策略;在传粉过程中,2种雄蕊既有分工,又有协作.2种雄蕊花药开裂时间不同步,花粉活力维持时间也不同,花粉的这种装配机制是紫薇提高花粉输出效率的繁殖策略.     

Effect of Pheromone Blend Components,Sex Ratio,and Population Size on the Mating of Cadra cautella (Lepidoptera: Pyralidae)

The almond moth Cadra cautella (Walker), a key pest of storage facilities, is difficult to manage using synthetic chemicals. Pheromone-based management methods remain a high priority due to advantages over conventional management practices, which typically use insecticides. Cadra cautella females release a blend of pheromone including (Z, E)-9,12-tetradecadienyl acetate (ZETA) and (Z)-9-tetradecadien-1-yl acetate (ZTA). The effect of these components on mating of C. cautella and how response varies with the population density and sex ratio remain unknown. In this study, the mating status of C. cautella was studied inside mating cages under different ratios of ZETA and ZTA diluted in hexane and at different population sizes either with equal or unequal sex ratio. The lowest percentage of mated females (highest mating disruption [MD] effects), corresponding to roughly 12.5%, was produced by a 5:1 and 3.3:1 ratio of ZETA:ZTA. Populations with equal sex ratio showed the lowest percentage of mated females, at 20% and 12.5% under lower and higher density, respectively. The next lowest percentage of mated females was produced when the sex ratio was set to 1: 2 and 2:1 male:female, with just 25% and 22.5% of moths mated, respectively. This study shows that mating status of C. cautella is influenced by ZETA:ZTA ratio, sex ratio, and population size. This current knowledge would have useful implications for mating disruption programs.     

High vegetative compatibility diversity of Cryphonectria parasitica infecting sweet chestnut (Castanea sativa) in Britain indicates multiple pathogen introductions

Chestnut blight, caused by Cryphonectria parasitica, was identified in Devon, UK, in December 2016. Intensive surveys detected the disease at further sites in Devon (seven), Berkshire (one), Dorset (one), Derbyshire (four) and a cluster of eight sites in southeast London. Over 570 survey samples were tested, and 227 were positive for C. parasitica by isolation and real-time PCR. A total of 227 isolates were tested for mating type, and 197 screened for vegetative compatibility group (VCG) and compared with VCGs known from mainland Europe. The same isolates were also screened for the presence of Cryphonectria hypovirus 1 (CHV-1). Eleven VCGs were identified within the UK population. Five corresponded to already known European VCGs but six were unique. The European VCGs mainly came from the Devon, Dorset, Berkshire and Derbyshire disease outbreaks, whilst unique VCGs were almost exclusively from the southeast London cluster. Both mating types were detected, but only one mating type was present at each site, with the exception of a single Devon site. Perithecia of C. parasitica were never observed at any site. CHV-1 was found in seven isolates from three different locations and was always subtype-I, which has limited hypovirulence. Therefore, although CHV-1 is associated with C. parasitica at some outbreaks, it probably has limited impact on virulence. The diversity of VCGs and their distribution at outbreak sites, together with findings of CHV-1, suggests C. parasitica has been introduced to the UK multiple times over at least two decades through international plant trade.     

Modified full-sib selection and best linear unbiased prediction of progeny performance in a European F2 maize population

Four cycles of modified recurrent full‐sib (FS) selection were conducted in an intermated F₂ population of European flint maize. The objectives of our study were to monitor trends across selection cycles in the estimates of population mean, inbreeding coefficients and variance components, and to investigate the usefulness of best linear unbiased prediction (BLUP) of progeny performance under the recurrent FS selection scheme applied. We used a selection rate of 25% for a selection index, based on grain yield and dry matter content. A pseudo‐factorial mating scheme was used for recombination. In this scheme, the selected FS families were divided into an upper‐ranking group of parents mated to the lower‐ranking group. Variance components were estimated with restricted maximum likelihood (REML). Average grain yield increased 1.2 t/ha per cycle, average grain moisture decreased 20.1 g/kg per cycle, and the selection index relative to the F₂ check entries decreased 0.3% per cycle. For a more precise calculation of selection response, the four cycles should be tested together in multi‐environmental trials. We observed a significant decrease in additive variance in the selection index, suggesting smaller future selection response. Predictions of FS family performance in Cn + 1 based on mean performance of parental FS families in Cn were of equal precision as those based on the mean additive genetic BLUP of their parents, and corresponding correlations were of moderate size for grain moisture and selection index.     

The frequency of natural cross-fertilisation in sequential sowings of winter barley

Summary Estimations of random mating frequency were computed for a series of sequential autumn sowings of populations of winter barley. The estimations were by means of the maximum likelihood scoring method and three varieties, each carrying a recessive genetic marker, provided three independent estimations for each population. High levels of out-crossing were found in sowings made early in September, and a trend towards absence of out-crossing in populations sown in late November was evident. Although fluctuations in this trend could be accounted for by fluctuations in meteorological factors, a more profound underlying effect was evident in that the three marker varieties behaved similarly, though to different degrees. It is postulated that the principle effect of sowing date, as it affects out-crossing, is upon the development of the flowering apex, and that environmental conditions at flowering time merely modify flowering behaviour.