Molecular characterization of strawberry pathogen Gnomonia fragariae and its genetic relatedness to other Gnomonia species and members of Diaporthales

Gnomonia fragariae is a poorly studied ascomycete belonging to Diaporthales. Originally G. fragariae was considered a saprophyte occurring on dead tissues of strawberry plants. Recently this fungus was found in Latvia and Sweden, and it was proven to be the cause of severe root rot and petiole blight of strawberry. Thirteen isolates of this pathogen and several other Gnomonia species occurring on rosaceous hosts were characterized by molecular analysis using nucleotide sequences of partial LSU rRNA gene and the total ITS region. The homologous regions from relevant diaporthalean taxa available in the GenBank were also included and compared with the taxa sequenced in this study. Phylogenetic analyses revealed that G. fragariae, G. rubi, and Gnomonia sp. (CBS 850.79) were genetically different from G. gnomon, the type species of the genus, and other members of Gnomoniaceae. The analyses showed that G. fragariae and Hapalocystis were genetically very closely related, forming a phylogenetic clade, which is possibly presenting a new family in the Diaporthales. Morphological comparisons of the Gnomonia species on the basis of commonly used criteria for the taxonomy of Diaporthales, so far did not reveal any evident features for the polyphyletic status of Gnomonia.     

Novel hosts of the Eucalyptus canker pathogen Chrysoporthe cubensis and a new Chrysoporthe species from Colombia

The pathogen Chrysoporthe cubensis (formerly Cryphonectria cubensis) is best known for the important canker disease that it causes on Eucalyptus species. This fungus is also a pathogen of Syzygium aromaticum (clove), which is native to Indonesia, and like Eucalyptus, is a member of Myrtaceae. Furthermore, C. cubensis has been found on Miconia spp. native to South America and residing in Melastomataceae. Recent surveys have yielded C. cubensis isolates from new hosts, characterized in this study based on DNA sequences for the ITS and β-tubulin gene regions. These hosts include native Clidemia sericea and Rhynchanthera mexicana (Melastomataceae) in Mexico, and non-native Lagerstroemia indica (Pride of India, Lythraceae) in Cuba. Isolates from these hosts and areas group in the sub-clade of C. cubensis accommodating the South American collections of the fungus. This sub-clade also includes isolates recently collected from Eucalyptus in Cuba, which are used to epitypify C. cubensis. New host records from Southeast Asia include exotic Tibouchina urvilleana from Singapore and Thailand and native Melastoma malabathricum (Melastomataceae) in Sumatra, Indonesia. Consistent with their areas of occurrence isolates from the latter collections group in the Asian sub-clade of C. cubensis. DNA sequence comparisons of isolates from Tibouchina lepidota in Colombia revealed that they represent a new sub-clade within the greater Chrysoporthe clade. Isolates in this clade are described as Chrysoporthe inopina sp. nov., based on distinctive morphological differences.     

The genus Setulipes (Marasmiaceae) in Madagascar and the Mascarenes, including a key to other African taxa

The authors report first records for the genus Setulipes in Madagascar, with the presence of Setulipes cf. hakgalensis and two new species, Setulipes funaliformis and S. moreaui, as well as a new species from Mauritius: S. mauritiensis. A key to these taxa, as well as to other African species, is supplied.     

The type species of Apiognomonia, A. veneta, with its Discula anamorph is distinct from A. errabunda

Species of Apiognomonia with their Discula anamorphic states in the Gnomoniaceae, Diaporthales, are known throughout the temperate Northern Hemisphere and cause diseases such as sycamore or plane tree anthracnose. The genus Apiognomonia was described based on A. veneta as the type species; however, there has been disagreement about whether or not A. veneta is a synonym of A. errabunda. Using morphological, ecological, and DNA sequence data we conclude that A. errabunda and A. veneta are different species, although very closely related; thus, A. veneta is the correct name for the type species of Apiognomonia. This conclusion is based on a combined analysis of sequences from the ITS regions of nuclear rDNA for 51 isolates from host plants of eight genera and intron regions from actin, calmodulin and translation elongation factor 1-alpha for over 25 isolates. The type species of the genus Discula is D. nervisequa, the earliest available epithet for D. platani, the lectotype of Discula. D. nervisequa is the anamorph of A. veneta. Based on an examination of the type specimen, we determined that the commonly used name for the anamorph of A. errabunda, D. umbrinella, refers to another species. A. veneta and A. errabunda including their anamorphs are described and illustrated. An account of all synonyms and excluded synonyms is presented.     

Physiological acclimation to light in Chara intermedia nodes

In this study we investigated the ability of Chara intermedia to acclimate to different irradiances (i.e. “low-light” (LL): 20–30 μmol photons m⁻² s⁻¹ and “high-light” (HL): 180–200 μmol photons m⁻² s⁻¹) and light qualities (white, yellow and green), using morphological, photosynthesis, chlorophyll fluorescence and pigment analysis.Relative growth rates increased with increasing irradiance from 0.016 ± 0.003 (LL) to 0.024 ± 0.005 (HL) g g⁻¹ d⁻¹ fresh weight and were independent of light quality. A growth-based branch orientation towards high-light functioning as a mechanism to protect the plant from excessive light was confirmed. It was shown that the receptor responsible for the morphological reaction is sensitive to blue-light.C. intermedia showed higher oxygen evolution (up to 10.5 (HL) vs. 4.5 (LL) nmol O₂ mg Chl⁻¹ s⁻¹), photochemical and energy-dependent Chl fluorescence quenching and a lower Fv/Fm after acclimation to HL. With respect to qP, the acclimation of the photosynthetic apparatus depended on light quality and needed the blue part of the spectrum for full development. In addition, pigment composition was influenced by light and the Chl a/Car and Antheraxanthin (A) + Zeaxanthin (Z)/Violaxanthin (V) + A + Z (DES) ratios revealed the expected acclimation behaviour in favour of carotenoid protection under HL (i.e. decrease of Chl a/Car from 3.41 ± 0.48 to 2.30 ± 0.35 and increase of DES from 0.39 ± 0.05 to 0.87 ± 0.03), while the Chl a/Chl b ratios were not significantly affected. Furthermore it was shown that morphological light acclimation mechanisms influence the extent of the physiological modifications.     

Natural variation of ascospore and conidial germination by Fusarium verticillioides and other Fusarium species

Fusarium verticillioides and other Fusarium species were examined for their spore germination phenotypes. In general, germinating spores of F. verticillioides formed germ tubes that immediately penetrated into agar. Such invasive germination was the predominant growth phenotype among 22 examined field isolates of F. verticillioides from a broad range hosts and locations. However, two of the field isolates were unique in that they formed conidial germ tubes and hyphae that grew along the surface of agar before penetration eventually occurred. Conidia of 22 other Fusarium species were assessed for their germination phenotypes, and only some strains of F. annulatum, F. fujikuroi, F. globosum, F. nygamai, and F. pseudoanthophilum had the surface germination phenotype (21 % of the strains assessed). Sexual crosses and segregation analyses involving one of the F. verticillioides surface germination strains, NRRL 25059, indicated a single locus, designated SIG1 (surface vs. invasive germination), controlled the germ tube growth phenotypes exhibited by both conidia and ascospores. Perfect correlation was observed between an ascospore germination phenotype and the germination phenotype of the conidia produced from the resulting ascospore-derived colony. Recombination data suggested SIG1 was linked (7 % recombination frequency) to FPH1, a recently described locus necessary for enteroblastic conidiogenesis. Corn seedling blight assays indicated surface germinating strains of F. verticillioides were less virulent than invasively germinating strains. Assays also indicated pathogenicity segregated independently of the FPH1 locus. Invasive germination is proposed as the dominant form of spore germination among Fusarium species. Furthermore, conidia were not necessary for corn seedling disease development, but invasive germination may have enhanced the virulence of conidiating strains.     

中间锦鸡儿CiMYB68基因克隆及表达分析

该研究以中间锦鸡儿(Caragana intermedia)为材料,利用RACE技术克隆了CiMYB68基因的全长序列。对CiMYB68的基因组DNA和cDNA全长分析显示,CiMYB68基因无内含子,开放阅读框为852bp,编码284个氨基酸。预测CiMYB68基因编码的蛋白质等电点为8.95,分子量约为31 459.4Da。序列比对和系统进化分析表明,该蛋白和大豆的GmMYB68一致性最高,达到67%。构建了CiMYB68基因与GFP融合表达质粒,激光共聚焦显微镜观察发现,融合蛋白定位于细胞核。用实时荧光定量PCR技术对在不同胁迫条件下CiMYB68基因的表达检测结果表明,在干旱和低温处理下CiMYB68均受到不同程度的诱导,暗示CiMYB68基因可能与中间锦鸡儿响应逆境胁迫有关。     

中间锦鸡儿CiPUB22基因克隆与功能分析

中间锦鸡儿(Caragana intermedia)耐旱、耐寒、耐盐碱,是西北干旱地区的重要固沙灌木,筛选其优良抗逆境基因,可以作为林草基因工程的基因源。该研究在中间锦鸡儿干旱胁迫转录组文库中找到1条CiPUB22 (plant U box 22)基因的cDNA全长序列,CiPUB22基因包括1 260 bp开放阅读框,编码419个氨基酸。实时荧光定量PCR结果表明,在脱水、盐和ABA处理1 h后CiPUB22基因表达量上升并达到最高水平,分别为对照表达量的12倍、35倍和7倍,干旱处理后12 d达到最高值,为对照的2.5倍,表明CiPUB22的转录水平受非生物胁迫诱导。构建CiPUB22基因的过表达载体并转化野生型拟南芥,对转基因纯合体株系抗逆性分析发现,在150 mmol/L NaCl、1 μmol/L ABA和400 mmol/L甘露醇处理下,过表达株系的萌发率均低于野生型,说明过表达CiPUB22基因降低了拟南芥在种子萌发过程中对盐和渗透胁迫的耐受性。     

内蒙古高原小叶锦鸡儿与中间锦鸡儿遗传多样性的研究

利用ISSR分子标记,选取内蒙古高原自东向西共15个锦鸡儿种群(7个小叶锦鸡儿种群,8个中间锦鸡儿种群)进行遗传多样性分析。结果表明:(1)7个小叶锦鸡儿种群总的多态位点百分数为98.19%,Nei’s基因多样性指数与Shannon信息指数分别为0.289 7、0.444 0;居群间遗传分化系数Gst为0.119 0,表明小叶锦鸡儿种群存在中等程度的遗传分化;居群间基因交流频繁,基因流(Nm)为3.701 0。(2)8个中间锦鸡儿种群多态位点百分数为99.7%,Nei’s基因多样性指数为0.312 8,Shannon信息指数为0.478 4;居群间遗传分化系数(Gst)为0.188 1,表明中间锦鸡儿种群存在较大的遗传分化,基因流(Nm)为2.157 8。(3)中间锦鸡儿遗传多样性及遗传分化均高于小叶锦鸡儿,这是中间锦鸡儿替代小叶锦鸡儿适应不良环境扩展分布的遗传基础。(4)邻接聚类分析结果显示,15个锦鸡儿种群表现为从东到西逐渐聚类,体现了这2个种在分布上的地理渐变性及地理替代性;遗传多样性各指数与气象因子及土壤因子之间的相关性分析表明,平均气温对锦鸡儿种群的遗传分化起着重要的作用。     

中间锦鸡儿转录因子基因 CiNAC1的克隆及功能分析

NAC转录因子家族是植物特有的、最大的转录因子家族之一,在植物应答非生物胁迫和生长发育过程中有重要的功能。该研究通过PCR技术,克隆得到了中间锦鸡儿 CiNAC1基因1 066 bp的cDNA全长序列。生物信息学分析显示, CiNAC1基因的开放阅读框(ORF)为921 bp,编码306个氨基酸,推导的蛋白分子量为34.57 kD,等电点为8.35,是一种亲水性蛋白,N端具有保守的NAM结构域,具有26个磷酸化位点和7个糖基化位点。实时荧光定量PCR检测显示,中间锦鸡儿 CiNAC1基因表达受干旱、高盐、脱水、高pH诱导;亚细胞定位发现CiNAC1定位到细胞核中,这与它作为转录因子的功能是一致的;转CiNAC1基因拟南芥株系侧根数目显著多于野生型,根长也明显比野生型长。研究认为, CiNAC1基因可能与中间锦鸡儿响应逆境胁迫机制有关。     

荒漠草原中间锦鸡儿冠层截留特征

以荒漠草原人工中间锦鸡儿（Caragana intermedia）为研究对象,利用2016-2017年监测获取的26次降雨事件,对比分析了两组灌丛（自然组和人工组）的冠层截留特征。结果表明：（1）试验期间共观测到有效降雨33次,总降雨量为251 mm,次平均降雨量和降雨强度分别为7.6 mm和1.14 mm/h,以雨量<2 mm,雨强<1 mm/h和降雨历时2-5 h的降雨出现次数最多;（2）自然组和人工组中间锦鸡儿平均截留量分别为1.11 mm和0.72 mm,平均截留率分别为24.81%和15.95%,两组灌丛截留存在极显著差异（P < 0.01）;（3）在雨量级>15 mm时,自然组（4.57%,CV=73.38%）和人工组（5.25%,CV=51.96%）平均截留率变异性相差最大;（4）自然组和人工组截留量与降雨量,降雨历时和降雨强度之间的关系可以用幂函数描述,截留率与三者的关系均用指数函数描述较好。在降雨特征相同的情况下,灌丛形态特征是影响中间锦鸡儿冠层截留的关键因素。     

New fungal genera, Tectonidula gen. nov. for Calosphaeria-like fungi with holoblastic-denticulate conidiogenesis and Natantiella gen. nov. for three species segregated from Ceratostomella

Two morphologically similar groups of ascomycetes with globose to subglobose perithecia, elongate necks, unitunicate asci floating freely at maturity, and hyaline ascospores currently placed in Calosphaeria s. lat. and Ceratostomella s. lat., respectively, are studied. The Calosphaeria-like fungi have groups of perithecia growing between cortex and wood, arranged in circular groups with converging necks and piercing the cortex in a common point; the asci with a shallow apical ring and U- to horseshoe-shaped hyaline ascospores are compared with Calosphaeria pulchella, the type species of the genus. Conidiogenesis of the investigated Calosphaeria-like fungi is holoblastic-denticulate; ramichloridium-like and sporothrix-like conidiophores and conidia were formed in vitro. Ascospore and ascus morphology, structure of the ascal apex, ascogenous system, mode of conidiogenesis and the large subunit rRNA sequences of this group differ considerably from C. pulchella and both groups are unrelated. Thus a new genus, Tectonidula, is described with two accepted species, T. hippocrepida and T. fagi; they are separated by ascospore and ascus morphology and holoblastic-denticulate conidiogenesis from the core species of Calosphaeria. The placement of Tectonidula among perithecial ascomycetes is discussed. The relationship of Tectonidula with Barbatosphaeria and two ramichloridium-like hyphomycetous genera Rhodoveronaea and Myrmecridium is investigated. Three species formerly attributed to Ceratostomella are studied. The revision of the herbarium type specimen and fresh material of Ceratostomella ligneola revealed that it is conspecific with Ceratostomella ampullasca and Ceratostomella similis. The LSU phylogeny clearly separated C. ligneola from Ceratostomella s. str. and morphologically similar Lentomitella. On the basis of molecular sequence data and detailed comparison of morphology of asci, ascospores and ascogenous system the genus Natantiella is described for C. ligneola with C. ampullasca and C. similis as its synonyms. Natantiella produced sterile mycelium in vitro.     

苏云金芽胞杆菌幕虫亚种晶胞粘连现象与晶体蛋白基因cry26Aa所在质粒有关

苏云金芽胞杆菌幕虫亚种T02菌株的伴胞晶体在芽胞外壁内侧形成,呈现晶胞粘连的现象。在此菌株中克隆了cry26Aa和cry28Aa两个基因,并对晶胞粘连现象与质粒的相关性做了系统研究。通过消除幕虫亚种T02菌株的质粒,得到了仅消除cry26Aa所在质粒的菌株BMB1151和无质粒的菌株BMB1152。通过穿梭载体将cry26Aa和cry28Aa两个基因分别和同时转化无质粒突变株BMB1152并表达,形成的晶体与芽胞独立存在不能粘连,表明在幕虫亚种染色体背景下仅仅cry的表达不能形成晶胞粘连现象,从而推断晶胞粘连现象可能与幕虫亚种两个基因所在的质粒有关;进一步的研究发现将cry26Aa在仅消除cry26Aa所在质粒的突变株BMB1151中表达,形成的晶体与芽胞也分别独立存在不能粘连,从而进一步推断幕虫亚种晶胞粘连现象与cry26Aa所在质粒有关。     

Synthesis of -Dopa by Escherichia intermedia cells immobilized in a carrageenan gel

Escherichia intermedia cells were immobilized by entrapment in a carrageenan gel and used for -DOPA synthesis from catechol, pyruvate, and ammonia. A preparation containing 75 mg of cell per gram of gel retained 60–65% of its original activity. The effect of substrate concentrations on the initial rate of -DOPA synthesis was very similar for free and immobilized cells, and substrate inhibition was observed for the three substrates. In batch reactors, up to 7.8 g l⁻¹ of -DOPA was obtained in 20 h (productivity 0.39 g l⁻¹ h⁻¹). Cells immobilized in a carrageenan gel showed higher -DOPA synthesis, in both initial rates conditions and batch reactors, than cells immobilized in a polyacrylamide gel.     

蝇蛹俑小蜂对橘小实蝇和瓜实蝇的偏好性

【背景】蝇蛹俑小蜂是实蝇类害虫蛹期的一种重要寄生蜂,对压制下一代实蝇类害虫的种群数量具有重要作用,但有关其对不同实蝇害虫的寄生特性尚缺乏研究。【方法】采用"H"型装置和培养皿测定方法,研究了蝇蛹俑小蜂的寄主选择偏好性。【结果】蝇蛹俑小蜂在橘小实蝇蛹和瓜实蝇蛹共存的情况下,偏好在橘小实蝇蛹上停留,且寄生率较高,最高寄生率达61.11%。【结论与意义】本研究为合理利用蝇蛹俑小蜂控制实蝇类害虫提供了理论基础。     

Two new Fusicoccum species from Acacia and Eucalyptus in Venezuela, based on morphology and DNA sequence data

Botryosphaeria spp. are common endophytes of woody plants, and they also include some serious pathogens of Eucalyptus and Acacia species. Numerous anamorphs have been associated with Botryosphaeria, of which the species Fusicoccum are amongst the most common. Here, we characterize two new Fusicoccum species, isolated from Eucalyptus and Acacia trees in Venezuela, based on morphological features in culture and comparisons of DNA sequence data. The two taxa named Fusicoccum andinum and F. stromaticum spp. nov, reside in two well-supported clades (BS values = 100 %) based on a combined data set of the ITS of the rDNA operon and translation elongation factor 1-α (EF1- α) gene sequences. The conidia of F. andinum are unusually large amongst Botryosphaeria anamorphs, and peripherally resemble those of B. mamane and B. melanops. F. stromaticum is characterized by large conidiomata in cultures, growth at 35 °C and slightly thickened conidial walls, which is different to most other Fusicoccum spp. No teleomorphs were observed for these fungi, but DNA sequence data show that they are anamorphs of Botryosphaeria.     

Phylogeny of Caragana (Fabaceae) based on DNA sequence data from rbcL, trnS–trnG, and ITS

Phylogenetic relationships of 48 species of Caragana (Fabaceae: tribe Hedysareae) and one representative each of Astragalus, Calophaca, Halimodendron, and Hedysarum are estimated from DNA sequences of the rbcL gene, trnS–trnG intron and spacer, and ITS region. At least one representative of all five sections and 12 series within Caragana are included. Analyses yielded strongly supported clades corresponding to sections Caragana, Bracteolatae, and Frutescentes. The species of section Jubatae are distributed among three strongly supported clades, i.e., one with the species of section Bracteolatae, another with two species of section Spinosae, and a third as sister to section Frutescentes. All but the last of these six clades are corroborated by at least one unambiguously traced morphological character. The placement of the other four species of section Spinosae are not well supported and lack unambiguous morphological synapomorphies, and the samples of Calophaca and Halimodendron nest within Caragana with weak support.