Mitochondrial DNA Variability in Fusarium Proliferatum (Gibberella Intermedia)

Restriction fragment length polymorphisms (RFLP) were used to assess genetic diversity of mitochondrial DNA (mtDNA) among 184 isolates of Fusarium proliferatum recovered from maize, asparagus, palms and reed. All strains were cross-fertile with standard mating type tester strains of Gibberella intermedia. Sixteen mitochondrial haplotypes were identified following digestion of DNAs with HaeIII, with seven, seven, five and six different haplotypes from maize, asparagus, palms and reed, respectively. Four haplotypes (I, III, IV and VII) were found on more than one host. Of these four, haplotype I was dominant on maize, representing 71% of the isolates. The banding patterns for haplotypes III and IV were >90% similar to the banding pattern of haplotype I. Haplotypes I, III and IV accounted for 87% of the isolates from maize, but were less common on the other hosts, accounting for 70%, 52% and 33% of the isolates from asparagus, palms and reed, respectively. Thirteen of the 16 haplotypes were recovered from only a single host plant species. When comparing the banding patterns and frequencies of these haplotypes, at least five were recovered at a higher frequency from one host relative to the others. Our results suggest that mtDNA RFLP analysis is a useful indicator of genetic divergence in Fusarium proliferatum.     

Susceptibility of Douglas fir (Pseudotsuga menziesii) to pitch canker, caused by Gibberella circinata (anamorph = Fusarium circinatum)

For better characterization of the risk of pitch canker (caused by Gibberella circinata , anamorph =  Fusarium circinatum ) to Douglas fir ( Pseudotsuga menziesii ), Californian isolates, selected exotic isolates, and ascospore progeny of a cross between wild-type Californian isolates were tested for aggressiveness to this host species. In addition, seedlings from representative provenances of P. menziesii in California were tested for susceptibility to pitch canker. The results revealed only minor differences between isolates, but differences in susceptibility between trees were often significant. The majority of the tested trees were relatively resistant as indicated by the development of only very short lesions, but some were clearly susceptible.     

Genetic Mapping of Pathogenicity and Aggressiveness of Gibberella zeae (Fusarium graminearum) Toward Wheat

ABSTRACT Gibberella zeae is the major fungal pathogen of Fusarium head blight of wheat and produces several mycotoxins that are harmful to humans and domesticated animals. We identified loci associated with pathogenicity and aggressiveness on an amplified fragment length polymorphism based genetic map of G. zeae in a cross between a lineage 6 nivalenol producer from Japan and a lineage 7 deoxynivalenol producer from Kansas. Ninety-nine progeny and the parents were tested in the greenhouse for 2 years. Progeny segregated qualitatively (61:38) for pathogenicity:nonpathogenicity, respectively. The trait maps to linkage group IV, which is adjacent to loci that affect colony pigmentation, perithecium production, and trichothecene toxin amount. Among the 61 pathogenic progeny, the amount of disease induced (aggressiveness) varied quantitatively. Two reproducible quantitative trait loci (QTL) for aggressiveness were detected on linkage group I using simple interval analysis. A QTL linked to the TRI5 locus (trichodiene synthase in the trichothecene pathway gene cluster) explained 51% of the variation observed, and a second QTL that was 50 centimorgans away explained 29% of the phenotypic variation. TRI5 is tightly linked to the locus controlling trichothecene toxin type. The two QTLs, however, were likely part of the same QTL using composite interval analysis. Progeny that produced deoxynivalenol were, on average, approximately twice as aggressive as those that produced nivalenol. No transgressive segregation for aggressiveness was detected. The rather simple inheritance of both traits in this interlineage cross suggests that relatively few loci for pathogenicity or aggressiveness differ between lineage 6 and 7.     

Mitochondrial DNA Diversity and Lineage Determination of European Isolates of Fusarium graminearum (Gibberella zeae)

Restriction fragment length polymorphisms (RFLPs) were used to assess genetic diversity of mitochondrial DNA (mtDNA) among standard isolates of seven lineages of Fusarium graminearum. The mtDNA patterns within each lineage were very similar (>89%), whereas significant differences were observed between the isolates belonging to different lineages, with the exception of lineages 1 and 4 where strong similarity was found between the RFLPs. Analysis of different band patterns resulted in characteristic HhaI and HaeIII bands that were suitable for identification of members of lineages 7, 6, 5, 3 and 2. Investigation of lineage distribution of 144 European isolates revealed that 142 belong to lineage 7. These data, therefore, confirmed the hypothesis that members of lineage 7 are predominant in Europe. Further analysis of isolates belonging to lineage 7 resulted in five haplotypes. These haplotypes have arisen as different combinations of three RFLP patterns for both HaeIII and HhaI restriction enzymes. Two isolates from Hungary, however, shared the same mtDNA RFLP profiles with a standard isolate of lineage 3, indicating that members of lineage 3, at a lower frequency, may also occur in Europe.     

广西稻恶苗病发生历史概况分析

稻恶苗病在广西属普遍发生的水稻常见病之一，其仅在少数年份的局部地区，个别品种上达到偏重发生程度，建国50天来，属偏重发生年的有6天，其中以1982年发生最重，属偏轻发生年的有34天，20世纪60年代和90年代全部属偏轻发后年份，笔者运用时间序列分析法的ARIMA（3，1，3）模型模拟预测2000年后5天稻恶苗病的发生趋势，结果提示今后5天该病的发生将有一个明显上升过程。     

Vegetative Compatibility and Mycotoxin Chemotypes among Fusarium graminearum (Gibberella zeae) Isolates from Wheat in Argentina

Gibberella zeae (anamorph Fusarium graminearum) is the main pathogen causing Fusarium head blight of wheat in Argentina. The objective of this study was to determine the vegetative compatibility groups (VCGs) and mycotoxin production (deoxynivalenol, nivalenol and 3-acetyl deoxynivalenol) by F. graminearum populations isolated from wheat in Argentina. VCGs were determined among 70 strains of F. graminearum isolated from three localities in Argentina, using nitrate non-utilizing (nit) mutants. Out of 367 nit mutants generated, 41% utilized both nitrite and hypoxanthine (nit1), 45% utilized hypoxanthine but not nitrite (nit3), 9% utilized nitrite but not hypoxanthine (NitM) and 5% utilized all the nitrogen sources (crn). The complementations were done by pairing the mutants on nitrate medium. Fifty-five different VCGs were identified and the overall VCG diversity (number of VCGs/number of isolates) averaged over the three locations was 0.78. Forty-eight strains were incompatible with all others, thus each of these strains constituted a unique VCG. Twenty-two strains were compatible with other isolates and were grouped in seven multimembers VCGs. Considering each population separately, the VCG diversity was 0.84, 0.81 and 1.0 for San Antonio de Areco, Alberti and Marcos Juarez, respectively. Toxin analysis revealed that of the 70 strains of F. graminearum tested, only 90% produced deoxynivalenol, 10% were able to produce deoxynivalenol and very low amounts of 3-acetyldeoxynivalenol. No isolate produced nivalenol. The results indicate a high degree of VCG diversity in the F. graminearum populations from wheat in Argentina. This diversity should be considered when screening wheat germplasm for Fusarium head blight resistance.     

上海地区水稻恶苗病的回升因素与防治对策

水稻恶苗病俗称白秆病、抢先稻。多年来自然发病较重。据近年各区（县）植保部门调查统计，恶苗病从秧苗期至抽穗期都有发生，自然发病株率在秧苗期为５％～１０％，在拔节孕穗期为１０％～２０％，严重田块病株率达３０％以上。通过严格的种子处理后，一般品种病株率在０．３％左右，感病品种为２％左右。而２００１年在局部地区水稻恶苗病发生很重，部分田块发病株率达１５％以上。为此，作者与基层植保员进行了认真的走访、询问和调查工作，现将结果简述如下。１发生现状根据市、区（县）有关部门于２００１年８月下旬至９月中旬对６个区…     

Genetic Variability and Population Structure of the Wheat Foot Rot Fungus, Fusarium culmorum, in Tunisia

The random amplified polymorphic DNA (RAPD) method was used to investigate the genetic variability and population structure of Fusarium culmorum isolated from wheat stem bases. A total of 108 isolates, representing seven geographically distinct populations, was collected from five climatic regions in Tunisia. Pseudo-allelic frequencies were estimated at each of the 25 putative RAPD loci analyzed by scoring for the presence or absence of amplified fragments; 92 haplotypes were found among the 108 strains. The analysis of the population structure did not reveal any trend with regard to geographic origin. Total gene diversity (H_T ^* = 0.318) was mostly attributable to diversity within populations (H_S ^* = 0.308). Analysis of molecular variance confirmed that most of the genetic variability was within populations. Genetic differentiation among populations was low to moderate (G_ST ^* ranged from 0 to 0.190 and averaged 0.041 over all loci). Cluster analysis with UPGMA using genetic distances did not reveal any spatial clustering of the isolates collected from the different geographic regions. Based on these results, we conclude that the F. culmorum isolates recovered from different regions in Tunisia might be part of a single population pool.     

糜子抗旱及复水相关基因的cDNA-AFLP差异显示

以糜子抗旱种质为实验材料,采用cDNA-AFLP技术分析四叶期糜子幼苗叶片在干旱、复水与对照等条件下的基因表达差异。选取了36个引物组合(6个荧光标记TaqⅠ引物×6个非荧光标记MseⅠ引物)进行选择性扩增,结果表明,这些引物组合都有很好的扩增效果,在三种处理之间均能扩增出表达模式不同的差异片段。随机选取了8个干旱特有和4个复水特有的差异片段进行了克隆测序。利用NCBI的Blastn进行序列的dbEST比对结果表明,12个EST序列中有2个分别与小麦、玉米的EST序列有较高的相似性,其余10个与已知EST序列的同源性都很低,可能为新的糜子抗旱相关基因。用Blastx与Genebank的非冗余蛋白数据库进行比较结果表明,其中的一个序列(DR007245)与N-乙酰葡萄糖胺-N-乙酰胞壁酸五肽(UDP-N-acetylglucosamine--N-acetylmuramyl-(Pentapeptide))的同源性较高,两个序列(DR007249,DR007250)与水稻的反转录转座子蛋白具有较高的同源性,反转录转座蛋白在植物抗旱、耐盐等抗逆境方面有重要作用。另外还有两个序列(DR007251,DR007252)与两种假定蛋白同源性较高。     

Assessment of Subtractive Hybridization to Select Species and Subspecies Specific DNA Fragments for the Identification of Xylophilus ampelinus by Polymerase Chain Reaction (PCR)

Eighteen Bsp143I digested DNA fragments specific to Xylophilus ampelinus were cloned from a library enriched for X. ampelinus obtained after a subtractive hybridization step. It was also possible to clone specific DNA sequences directly after DNA digestion with Bsp143I probably because X. ampelinus is a unique bacterium. Nucleotidic sequences of four cloned specific fragments were determined. They did not share any homology with other DNA sequences in the EMBL/GeneBank database. Four primer sets were designed and tested for specificity to X. ampelinus. One primer set (Xamp 1.27) was a good candidate for a species-specific reagent in a procedure of identification of X. ampelinus using PCR. One primer set detected only Greek strains isolated from Vitis vinifera cv. Sultana. Genetic diversity within the X. ampelinus species can be used in further epidemiological studies on the bacterial necrosis of grapevine.     

Identification of Pathogenic Races 0, 1B/C, 5, And 6 Of Fusarium Oxysporum F. Sp. Ciceris With Random Amplified Polymorphic DNA (RAPD)

Ninety-nine isolates of Fusarium oxysporum f. sp. ciceris (Foc), representative of the two pathotypes (yellowing and wilt) and the eight races described (races 0, 1A, 1B/C, 2, 3, 4, 5, and 6), were used in this study. Sixty isolates were analyzed by the RAPD technique using DNA bulks for each race and 40 primers. Bands presumably specific for a DNA bulk were identified and this specificity was confirmed by further RAPD analysis of individual isolates in each DNA bulk. Primers OPI-09, OPI-18, OPF-06, OPF-10, and OPF-12 generated RAPD marker bands for races 0, 1B/C, 2, 3, 4, 5, and 6. The reliability and utility of this procedure was validated in blind trials using 39 new Foc isolates. Ten of the 39 isolates had already been typed to race by pathogenicity tests and 29 were typed both by pathogenicity and RAPD testing in this study. In these blind trials, we assigned the 39 new isolates to a race solely on the basis of their RAPD haplotype. Thus, we concluded that Foc races 0, 1B/C, 5, and 6 can be characterized by the RAPD markers. Cluster analysis of the RAPD data set resulted in three clusters of isolates within Foc. The yellowing isolates were grouped in two distinct clusters which correspond to races 0 and 1B/C. The wilt isolates constitute a third cluster that included races 1A, 2, 3, 4, 5, and 6. These results provide a means of studying the distribution of Foc races, to assist in the early detection of introduced race(s) and to facilitate the efficient deployment of available host resistance.     

杀菌剂与吡虫啉混合配剂对褐飞虱种群增长的影响

褐飞虱Nilaparvata lugens Stål是亚洲国家水稻产区的主要害虫,其腹部脂肪体内存在大量类酵母共生菌（Yeast-like symbiotes,YLS）。YLS对褐飞虱的生长发育和繁殖起着至关重要的作用。以YLS为靶标,通过抑制褐飞虱体内YLS从而控制褐飞虱的发生必将是今后有效防治褐飞虱的新途径。前期研究表明,与单一的化学杀虫剂相比,杀虫剂和杀菌剂的混配制剂因能有效抑制褐飞虱体内YLS的数量从而导致褐飞虱更高的死亡率。本研究利用生命表和Morris-Watt数学模型分析了27%丰加霉素&四霉素P&四烯菌素B&四霉素A和75%肟菌酯&戊唑醇分别与吡虫啉的混合配剂对褐飞虱种群增长影响。结果表明,2种混合配剂处理后褐飞虱体内YLS的数量均明显下降,且褐飞虱种群增长指数显著低于吡虫啉处理组,混合配剂对褐飞虱的抑制作用主要表现在卵孵化期和成虫产卵期。本研究结果为褐飞虱防治提供了新的思路和方法。     

基于两性生命表的以朱砂叶螨为猎物的加州新小绥螨四川种群生长发育、繁殖及捕食率

为全面掌握加州新小绥螨四川种群的种群特征,本试验运用两性生命表开展了以朱砂叶螨为猎物的加州新小绥螨四川种群的生长发育、繁殖与捕食率研究。结果表明,加州新小绥螨四川种群不同发育阶段重叠,在雌成螨产卵期结束前雌成螨特定年龄-阶段存活率S_xj明显高于雄成螨。单位时间（12 h）特定年龄-阶段繁殖力f_x5、特定年龄繁殖力m_x最大值分别为1.74和1.12粒/雌,单雌总产卵量为49.28粒。种群内禀增长率为0.27/d、周限增长率为1.31/d、净增殖率为26.61粒/个体、世代平均周期为12.34 d。在未成熟阶段,加州新小绥螨各螨态特定年龄-阶段捕食率C_xj均呈双峰型,雌成螨和雄成螨捕食率均随时间明显波动。加州新小绥螨取食朱砂叶螨的转化率为4.55、内禀捕食率为0.54、周限捕食率为1.72头。以朱砂叶螨为猎物时,加州新小绥螨四川种群表现出较好的个体发育、种群增长和捕食能力,可作为控制朱砂叶螨的生防资源,值得进一步研究和开发利用;同时朱砂叶螨可作为加州新小绥螨四川种群人工繁殖选择的猎物之一。     

Photodynamic oxygen activation by rubellin D,a phytotoxin produced by Ramularia collo-cygni (Sutton et Waller)

Ramularia collo-cygni, a pathogenic deuteromycete, is the causal agent of Ramularia leaf spot disease on barley. It produces a number of coloured metabolites of which one was identified as rubellin D, a phytotoxin only described for Mycosphaerella rubella until now. Rubellin D induces light- and concentration-dependent necrosis on barley leaves. In a biochemical model system we could demonstrate that rubellin D exhibits photodynamic activity triggering the light-dependent production of reactive oxygen species (ROS). The use of inhibitors of ROS production confirmed mainly the production of singlet oxygen and to a lesser extent superoxide. In addition, light-dependent peroxidation of α-linolenic acid by rubellin D was shown. Based on our results we present a model for ROS-mediated symptom induction on barley leaves by Ramularia collo-cygni.     

Evidence of Genetic Exchange by Parasexual Recombination and Genetic Analysis of Pathogenicity and Mating Type of Parasexual Recombinants in Rice Blast Fungus, Magnaporthe oryzae

ABSTRACT A selectable marker gene conferring resistance to bialaphos (BI) was introduced into rice blast isolate Y90-71BI and another conferring resistance to blasticidin S (BS) into isolate 3514-R-2BS of Magnaporthe oryzae to demonstrate exchange of DNA. Colonies obtained from co-cultures of these two isolates were resistant to both BI and BS and had both resistance genes as shown by Southern blot analysis of their genomic DNA. Conidia from these BI-BS-resistant isolates had only one nucleus per cell after staining with 4',6-diamidino-2-phenylindole (DAPI). Using flow cytometry, however, these BI-BS-resistant isolates were found to be haploid. Segregation of BI-BS-resistant isolates for pathogenicity (avirulence to virulence) on rice line K59-1 was consistent with a 1:1 ratio, as was segregation for mating type. These BI-BS-resistant isolates were thus apparently derived from parasexual exchange of DNA and the segregation of pathogenicity and of mating type of the parasexual recombinants might correspond to that of the progeny of the offspring of the sexual cross.     

江西亚洲柑橘木虱的种群遗传结构研究

亚洲柑橘木虱Diaphorina citri Kuwayama是芸香科Rutaceae植物新梢期的重要害虫,也是黄龙病Huanglongbing的主要传播媒介,严重威胁江西柑橘产业的发展。本文利用线粒体基因标记,研究江西10个地点亚洲柑橘木虱的种群遗传结构和多态性。扩增了2个线粒体COⅠ基因片段,长度为1 431 bp。其中多态性位点2个(均为非同义突变,A/G转换),单倍型3个(Hap-1,Hap-2和Hap-3)。最常见的单倍型是Hap-1(占总样本数的91.7%),发生在所有采样点;其次为Hap-3(6.3%),分布在大余和崇义;Hap-2(2.0%)仅在兴国出现。单一直线型的单倍型网络结构模式、小而不显著的HR(Harpending’s raggedness)指数值,以及单峰型错配分布曲线均表明:江西地区的亚洲柑橘木虱是近期入侵并经历了快速的种群动态扩张。     

油茶象种群密度及空间格局

油茶象Curculio chinensis Chevrolat是我国特有木本油料树种——油茶Camellia spp.的专性蛀果害虫,常导致其大量落果。本文旨在明确油茶象幼体(卵+幼虫)种群动态、空间格局及其关键影响因子,以便为调查取样、长期监测和科学防控等提供理论依据。2017年在湖南常宁普通油茶林,2016年-2017年在湖南益阳小果油茶林于雌虫产卵季节进行了调查,解剖收集的果实并记录其所含幼体数。用6种聚集度指标(m~*/m、I_δ、C、I、C_a、k)和Taylor幂法则、Iwao回归模型分析油茶象幼体的空间格局,并探讨其关键影响因子。调查和分析表明,雌虫于6月上、中旬开始产卵,幼体种群密度呈单峰变化,2016年和2017年高峰期分别在7月25日和7月18日。小果油茶林2016年、2017年高峰期种群密度分别为17.95±1.53和16.27±1.06幼体/样株,高于普通油茶林,分别是普通油茶林种群密度(8.16±1.23幼体/样株)的2.20倍和1.99倍,差异极显著(P0.001)。聚集度指标分析显示,油茶象在普通油茶林均呈聚集分布,在小果油茶林呈聚集分布或随机分布。回归模型表明,该虫在两种油茶林均呈聚集分布。Blackith种群聚集均数λ2,表明油茶象幼体聚集是由亲代雌虫本身的产卵行为与寄主果实特征综合作用的结果。给出了最适理论抽样数公式并绘制了3种抽样精度下最适抽样数随种群密度变化的曲线图。综上,初步明确了油茶象幼体在普通油茶林和小果油茶林的空间分布格局和种群动态,并探讨了其关键影响因子。试验结果可为该虫调查取样、长期监测和科学防控等提供理论依据。     

贵州喀斯特稻区黑肩绿盲蝽种群发生规律研究

为了摸清喀斯特稻区黑肩绿盲蝽Cyrtorhinuslividipennis的发生规律,2010年在贵州省6个具有代表性的稻区开展了该天敌田间种群消长的监测研究。结果表明：田间黑肩绿盲蝽的始见期集中在6月上、中旬,水稻生长前期虫量发生较少,后期发生量较大,发生高峰期一般在7月下旬一8月下旬。黑肩绿盲蝽与褐飞虱Nilaparvatalugens的种群发生存在同步性。水稻害虫防治的常用药剂啶虫脒、阿维菌素、吡虫啉和毒死蜱对黑肩绿盲蝽种群发生有明显的抑制作用。