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1.
本文系统研究了紫蓬山虫生真菌资源概况。共采集分离获得436株虫生真菌,鉴定为6属13种,其中优势种为环链棒束孢、蝉棒束孢、球孢白僵菌和玫烟色棒束孢,4种优势种占所有菌株的89.23%。对其虫生真菌季节变化和分布特点进行研究发现,虫生真菌数量丰富但物种多样性指数较低;夏季和晚春虫生真菌最为丰富,随着气温及降雨量的下降,菌株数量逐渐下降;不同季节环链棒束孢、蝉棒束孢和球孢白僵菌的数量波动大,但玫烟色棒束孢波动较小。环链棒束孢、蝉棒束孢和玫烟色棒束孢主要分布于阔叶林中,对单一生境的依赖性较强;而球孢白僵菌主要分布于针叶林中。本研究明确了紫蓬山虫生真菌的物种多样性和分布特点,为进一步开发生物农药奠定基础。  相似文献   

2.
球孢白僵菌有性世代培养及子代毒力稳定性鉴定   总被引:1,自引:0,他引:1  
在利用球孢白僵菌Beauveria bassiana有性态改良球孢白僵菌菌株的研究中,菌株的选择非常重要,其中利用交配型基因选择菌株是必要手段。而且虫生真菌的有性态能够使菌株间产生基因交流,促进其进化的过程。为更好地研究球孢白僵菌的遗传规律,以获得避免菌种衰退的方法,本研究建立了分别以柞蚕蛹为活体寄主和糙米培养基为基质的2种人工培养有性世代繁殖体系。结果表明,交配型为MAT1-1-1型和MAT1-2-1型混合型菌株BbDPSD5,经过自交在2种基质上均能够产生有性态。而具有2种单一交配型的菌株BbOFDH1-5和BbDPSD2进行配伍试验,BbOFDH1-5×BbDPSD2、BbOFDH1-5×BbDPSD5和BbDPSD2×BbDPSD5在2种基质上均没有有性态产生。对获得的有性态子囊孢子经无性繁殖产生的菌株进行r DNA-ITS片段以及交配型基因检测,并对测序结果进行比对分析。结果显示,扩增得到的r DNA-ITS序列和MAT1-2-1交配型基因与亲本序列同源性为100%,而没有检测到MAT1-1-1型。2种培养方式获得的结果完全一致。对有性态子代子囊孢子产生的株系进行毒力测试结果表明,所有子代菌株与原始菌株BbDPSD5的毒力差异不显著,子代交配型基因虽然缺失了MAT1-1-1,但是毒力没有衰退,未出现菌种退化现象。  相似文献   

3.
为明确黑龙江省采集自不同年份、不同地区的稻瘟病菌Magnaporthe oryzae的育性能力和交配型分布,采用2株标准菌株GUY11(MAT1-2)和KA3(MAT1-1)对2016—2017年黑龙江省西部、东部、中部3个地区经单孢分离的241株稻瘟病菌进行育性测定,并利用PCR技术对其交配型进行检测。结果表明,黑龙江省西部、东部、中部的241株稻瘟病菌中可育性菌株比例为11.62%,其中雌性菌株、雄性菌株、两性菌株分别占1.66%、4.56%和1.25%,不能判断其性别的未知菌株占4.15%。采集自不同地区、不同年份的稻瘟病菌可育性差异均较大,西部、东部、中部地区可育性菌株出现频率分别为13.25%、7.27%和12.62%;2016年采集的稻瘟病菌可育性较高,可育性菌株出现频率为25.30%。黑龙江省稻瘟病菌群体中同时存在MAT1-1和MAT1-2两种交配型,主要以交配型MAT1-1占优势,出现频率为58.92%,交配型为MAT1-2的菌株出现频率为8.30%。不同地区稻瘟病菌的交配型亦有差异,交配型为MAT1-1的菌株在黑龙江省东部地区出现频率最高,为72.73%,在中部、西部地区的出现频率次之,分别为61.17%和46.99%。表明黑龙江省水稻种植区的稻瘟病菌同时存在2种交配型菌株,其交配型存在丰富的多态性,但其可育性及交配型分布不均衡。  相似文献   

4.
本文通过分析两种不同交配型球孢白僵菌菌株(MAT1-1-1,I型;MAT1-2-1,II型)在玉米植株中的定殖效率和回收菌株交配型基因序列的变异,拟揭示球孢白僵菌在玉米植株内生定殖过程中玉米对不同交配型菌株的选择规律、不同交配型菌株对环境的适应性能力。试验分为4个接种组,接种菌株为I型交配型菌株(Bb OFDH1-5)(处理B5)、II型交配型菌株(Bb DPSD2)(处理B2)、两个菌株等量混合(处理B5+B2)、对照组(处理CK),处理组B5、B2、B5+B2接种孢子浓度均为108孢子/m L,对照组接种量为零。结果表明,处理B5、B2、B5+B2三组中均有球孢白僵菌在玉米叶片中定殖,其中,交配型II菌株处理组B2中内生定殖效率高,检测率为33.00%;混合处理组B5+B2中交配型II菌株比I型菌株回收比率高,回收比为12:10。交配型基因序列分析显示,交配型I菌株处理组B5和混合处理组B5+B2的回收菌株中I型交配型基因非编码区序列出现多个位点突变,编码区仅在3个位点出现突变;交配型II菌株处理组B2和混合处理组B5+B2中II型交配型基因序列均没有发生变异。以上结论说明球孢白僵菌在玉米植株定殖过程中,交配型II菌株Bb DPSD2定殖率较I型菌株Bb OFDH1-5高;玉米植株更倾向于选择II型交配型菌株作为内生菌株;交配型基因MAT1-1-1比MAT1-2-1更易发生突变表明I型菌株对自然环境的适应性更强。  相似文献   

5.
绿僵菌Metarhizium spp.是虫生真菌的主要类群之一,是当前世界上研究和应用最广的虫生真菌之一。由于绿僵菌能在自然土壤环境下长期存活,对地下害虫的控制具有明显优势。本研究从蛴螬僵虫上分离出一株寄生真菌,结合形态学和EF-1α基因5'端序列分析结果鉴定为黄绿绿僵菌Metarhizium flavoviride var. pemphigi。菌株培养的试验表明,乳糖有利于该菌株的菌丝生长,葡萄糖有利于该菌株产孢,KNO3为该菌株最适宜的氮源,在乳糖和KNO3为碳氮源时,C/N比为34:1有利于菌丝生长及产孢,添入微量元素Mn能促进产孢。在灭菌土中含5×106孢子/g时该菌株的对筛胸梳爪叩甲幼虫的校正累积死亡率为57.14%,幼虫逃逸率为4.76%。  相似文献   

6.
本文基于ITS-5.8S rDNA序列对来自国内外的玫烟色棒束孢不同菌株进行了系统发育及DNA多态性研究。系统发育分析表明,玫烟色棒束孢全部被试序列都分离为两个完全独立的支系,分别为聚在分支CladeⅠ-1中且支持率为94%的菌株和聚在分支CladeⅡ-1内且支持率为90%的菌株;为了确定上述2支系的真正分类地位,对所试玫烟色棒束孢与爪哇棒束孢菌株进行了多态性研究,获得8个单倍型,单倍型多样性Hd=0.65,核苷酸多样性Pi=0.04,以及2个不同的DNA序列保守区等。通过系统发育及多态性研究结果表明所试玫烟色棒束孢无论在种的水平上或假定的种群水平上,其遗传多样性都较大、分化明显,它们的差异已超出了种的边界,分支CladeⅠ-1中的菌株是典型的玫烟色棒束孢,而CladeⅡ-1中的玫烟色棒束孢菌株可能是爪哇棒束孢。  相似文献   

7.
实地调查了徂徕山林场松阿扁叶蜂越冬期幼虫感病死亡情况,并对引起幼虫感病死亡的致病菌分离纯化,进行了形态鉴定和分子鉴定。结果表明,松阿扁叶蜂越冬感病幼虫在自然状态下感病致死率可达8.9%~21.3%,其病原分离物可分为真菌菌株和细菌菌株两大类。其中,真菌感染致死率为67.8%,细菌感染致死率为32.2%。从松阿扁叶蜂死亡幼虫虫体中分离到5株细菌[嗜虫沙雷氏菌(Serratia entomophila)、河生肠杆菌(Enterobacter amnigenus)、鲍氏不动杆菌(Acinetobcter baumannii)、黏质沙雷菌(Serratia marcescens)、枯草芽孢杆菌(Bacillus subtilis)和2株真菌(粉质棒束孢(Isaria farinosa)、环链棒束孢(Isaria cateinannulata)]。其中,2株细菌菌株(黏质沙雷菌、枯草芽孢杆菌)和2株真菌菌株(粉质棒束孢、环链棒束孢)对松阿扁叶蜂幼虫的室内试验校正致死率为24.2%~68.9%;林间试验校正致死率为16.7%~62.4%。其中,环链棒束孢的校正死亡率为62.4%。  相似文献   

8.
稻曲菌交配型初探   总被引:2,自引:0,他引:2  
 稻曲菌(有性态:Villosiclava virens;无性态:Ustilaginoidea virens)有性繁殖产生的子囊孢子是水稻稻曲病可能的初侵染源之一,交配型基因座对真菌有性繁殖中的性别控制起着决定性作用。为进一步揭示稻曲菌的有性繁殖方式,本研究首次克隆了稻曲菌交配型基因mat1-1-1的α-结构域(α-domain)相应核苷酸序列,发现其与麦角菌科真菌Cordyceps militaris、Cor. bassianaClaviceps purpureamat1-1-1基因中α-结构域相应核苷酸序列同源性分别为61%、63%和68%;根据mat1-1-1和mat1-2-1部分基因序列设计特异性引物,并使用PCR方法检测了来源于3个异源子座的240个子囊孢子单孢菌株和50个田间菌株的交配型基因,结果显示稻曲菌mat1-1-1和mat1-2-1基因分别存在于不同菌株中;将具有相同或不同交配型基因的菌株配对接种,发现菌核通常产生在mat1-1-1和mat1-2-1基因型菌株配对接种产生的稻曲球上,其中大部分菌核可萌发产生子座,而菌株单独接种或具有相同交配型基因的菌株配对接种水稻后多数不能形成菌核;根据以上结果初步推断稻曲菌为异宗配合真菌。  相似文献   

9.
茶小绿叶蝉Empoasca spp.是我国最重要的茶树害虫,几乎对所有化学农药都产生了很高的抗药性,亟需开发经济有效的生物农药。本研究通过对从茶小绿叶蝉上分离的1株爪哇棒束孢Isaria javanicus和4株自其它叶蝉上分离的球孢白僵菌Beauveria bassiana的胞外蛋白酶活性测定,初选出活性较高的爪哇棒束孢和两株球孢白僵菌;对假眼小绿叶蝉低龄若虫的生物测定结果显示,爪哇棒束孢毒力最强,分别比两株球孢白僵菌高2.33~7.26倍和6.08~27.26倍,在1×108孢子mL 1浓度条件下第13 d的致死率达100%,LT50为5.9 d。通过随机区组设计进行的田间防治试验结果表明,使用爪哇棒束孢菌株配制的IJOD-1悬乳剂以7.5×1013孢子hm 2的剂量常量喷施,对假眼小绿叶蝉虫口的防治效果在第11 d时为50.32%;第25 d时追加1次放菌,在第55 d时的防治效果达68.40%。  相似文献   

10.
安徽歙县油茶林昆虫病原真菌物种多样性及其季节变化   总被引:1,自引:0,他引:1  
为研究安徽歙县油茶林的昆虫病原真菌群落多样性,于2016-2017年对油茶林内自然罹病的昆虫标本进行分离和鉴定。结果表明,分离的昆虫病原真菌共计132株,鉴定为6属10种。球孢白僵菌Beauveria bassiana、环链棒束孢Isaria cateniannulata和细脚棒束孢I.tenuipes为歙县油茶林昆虫病原真菌的优势类群,3种昆虫病原真菌的数量占所有菌株的75.76%。同时对昆虫病原真菌物种的多样性研究表明,其物种数、多度、多样性指数、均匀度等指标在一年中变化不大。不同季节昆虫病原真菌数量变化较大,夏季菌株最为丰富,随着气温下降及降雨量的减少,菌株数量逐渐下降。本研究结果可为进一步开发生物农药奠定基础。  相似文献   

11.
 有性生殖在真菌的生活史和进化过程中具有重要作用,而交配型基因是控制有性生殖的关键因子。前期研究发现稻曲病菌(Villosiclava virens)MAT1-2型菌株中包含MAT1-2-1MAT1-2-8两个交配型基因,但是它们如何调控稻曲病菌有性生殖依然不清楚。本文研究了它们在不同侵染和生长发育时期的表达模式和编码的蛋白结构特性。研究表明MAT1-2-1在侵染不同阶段一直下调表达;而MAT1-2-8在侵染早期(5 dpi)上调表达,在侵染后期下调表达。与营养菌丝阶段比较,MAT1-2-1MAT1-2-8在有性发育过程菌核形成、菌核萌发、子座原基形成和子座成熟4个阶段的表达量都是下降的,在菌核形成阶段表达量最低。生物信息学分析显示MAT1-2-1和MAT1-2-8具有磷酸化位点,为非分泌蛋白,无明显的跨膜结构域。蛋白同源比对分析表明MAT1-2-1与香柱菌(Epichloë typhina)的MAT1-2-1同源性最高,而MAT1-2-8与绿僵菌(Metarhizium)的MBR_08192蛋白同源性最高。进一步研究发现MAT1-2-1和MAT1-2-8能够互作,并分别主要定位在细胞核和细胞基质中。通过质谱技术鉴定到MAT1-2-1的一些候选互作蛋白,如假定Ran交换因子Prp20/Pim1(KDB12229.1)、假定rRNA处理蛋白Ebp2(KDB12923.1)及组蛋白H1(KDB12711.1)等。因此,以上结果为研究稻曲病菌交配型基因MAT1-2-1MAT1-2-8调控有性生殖的生物学功能奠定了基础。  相似文献   

12.
ABSTRACT Dothistroma septosporum and D. pini are the two causal agents of Dothistroma needle blight of Pinus spp. in natural forests and plantations. Degenerate primers amplified portions of mating type genes (MAT1-1-1 and MAT1-2) and chromosome walking was applied to obtain the full-length genes in both species. The mating-type-specific primers designed in this study could distinguish between the morphologically similar D. pini and D. septosporum and between the different mating types of these species. Screening of isolates from global collections of D. septosporum showed that only MAT2 isolates are present in Australian and New Zealand collections, where only the asexual form of the fungus has been found. In contrast, both mating types of D. septosporum were present in collections from Canada and Europe, where the sexual state is known. Intriguingly, collections from South Africa and the United Kingdom, where the sexual state of the fungus is unknown, included both mating types. In D. pini, for which no teleomorph is known, both mating types were present in collections from the United States. These results provided new insights into the biology and global distribution of two of the world's most important pine pathogens and should facilitate management of the diseases caused by these fungi.  相似文献   

13.
玉米大斑病菌两性交配型菌株的出现频率及其育性分析   总被引:1,自引:1,他引:0  
玉米大斑病菌属于异宗配合真菌,在自然界中存在A、a交配型菌株和Aa两性交配型菌株,为明确两性菌株的出现频率及其对有性生殖的影响,采用有性态诱导和交配型PCR鉴定的方法,对2011—2014年采集的野生菌株及室内诱导产生的有性杂交后代F1、F2代进行交配型组成鉴定,通过检测两性菌株子囊壳和子囊孢子的发育情况明确其有性生殖能力,结合RT-PCR技术对有性生殖过程中MAT1和MAT2基因的表达情况进行分析。结果表明,野生菌株和有性杂交后代中均存在两性交配型菌株,且出现频率相当,介于2.09%和6.25%之间;Aa两性菌株的育性与杂交时对应菌株的交配型组成相关,两性菌株与A交配型菌株或a交配型菌株杂交均可育,能产生成熟的子囊壳和子囊孢子,但其自交时败育,只产生子囊壳,不产生子囊孢子。两性菌株Aa自交和两性菌株Aa与A交配型或与a交配型菌株杂交时,Aa菌株中MAT1和MAT2基因表达量无明显差异,推测MAT基因在有性生殖产生子囊壳的过程中发挥了重要作用。  相似文献   

14.
Sexual reproduction in fungi is controlled by mating type genes, which are located at the MAT locus. In this study, we investigated the structure of this locus in the phytopathogenic fungus Phyllosticta citricarpa, the causal agent of citrus black spot disease. Despite intensive study, its sexual state has never been observed in single-spore culture. Through analysis of the genome sequences of two individual P. citricarpa isolates, the sequence of the DNA lyase gene was identified and, as previously reported in the literature, the mating type genes were located in the 3′ flanking region of this gene. The results suggested that P. citricarpa is heterothallic, owing to the exclusive presence of the MAT1–1 or MAT1–2 gene in individual strains. In order to characterize the MAT locus, we designed primers to amplify this region. P. citricarpa was found to have complete and apparently functional copies of MAT genes, containing α-1 and HMG domains, present in different isolates. In addition to MAT1–2-1 and MAT1–1-1 genes, the MAT1–1-4 gene was located in the 5′ flanking region of the MAT1–1-1 gene and the MAT1–2-5 gene was located in 5′ flanking region of the MAT1–2-1 gene. A multiplex PCR protocol was also developed to differentiate P. citricarpa idiomorphs, which can be used in distribution and incidence studies of mating type strains, in order to determine the occurrence of sexual reproduction and to facility crossing studies. Furthermore, in Brazil, the two idiomorphs occur in a 1:1 ratio, which is expected in sexually reproducing populations.  相似文献   

15.
Mating strategies contributing to a balance between inbreeding and outcrossing are understudied in all but a few model fungal pathogens. This study examined factors that influence the occurrence of the sexual state of Pyrenophora semeniperda. It was consistently found to have functional copies of the MAT1-1 and MAT1-2 idiomorphs essential for sexual reproduction, despite considerable polymorphism in both single nucleotide polymorphisms (SNPs) and number of 18-base minisatellite repeats. The two idiomorphs occurred at approximately equal frequencies across 25 populations on Bromus tectorum seeds in the western United States, suggesting maintenance of sexual reproduction. The putative mating system is described as facultative pseudohomothallism, with only one of the two MAT1 idiomorphs found in a nucleus. Unikaryotic strains of opposite mating type can potentially mate, as can nuclei of opposite mating type in a thallus that results from anastomosis between vegetatively compatible unikaryotic strains. Strains shown to be dikaryotic using simple sequence repeat (SSR) markers may contain either or both MAT1 idiomorphs. Most populations consist of a mixture of MAT1-1, MAT1-2 and MAT1-1/MAT1-2 genotypes. A possible constraint on recombination is the presence of multiple strain groups characterized by ITS haplotype. These are apparently vegetatively incompatible, as even dikaryotic strains are invariably composed of a single ITS haplotype. Different ITS haplotypes also have unique combinations of MAT1-1 and MAT1-2 allelic variants, suggesting that perhaps these strain groups are also sexually incompatible. Phylogenetic analysis using both genome-wide SNP/indel polymorphisms and SSR markers demonstrated genetic divergence among ITS haplotypes, supporting the hypothesis that these strain groups may represent cryptic species.  相似文献   

16.
Mating type genes of Verticillium dahliae, a wilt pathogen affecting many plant species, were identified to examine sexual recombination between Japanese pathotypes. We amplified a DNA sequence encoding high mobility group (HMG) box from V. dahliae using PCR. A cloned genomic DNA fragment included a sequence homologous to MAT1-2-1 gene. Despite that sequence's presence in all V. dahliae isolates we used, MAT1-1-1 (an opposite mating type gene) was never amplified. We concluded that V. dahliae is potentially heterothallic. Furthermore, sexual bias practically obviates sexual recombination between Japanese pathotypes. This report describes, for the first time, a mating type gene of phytopathogenic Verticillium.  相似文献   

17.
18.
Verticillium dahliae, a soilborne plant pathogen, causes wilt disease in many important crops. We reported previously that the mating type gene MAT1-2-1 is spread to isolates of this asexual fungus. However, we did not determine whether V. dahliae is homothallic or heterothallic because the opposite mating type gene, MAT1-1-1, had not been identified. In the present study, we identified the MAT1-1-1 gene from an isolate lacking MAT1-2-1 and the mating type idiomorphs of V. dahliae. Each isolate we tested contained either the MAT1-1 or MAT1-2 idiomorph, indicating that the asexual fungus V. dahliae is potentially heterothallic.  相似文献   

19.
小麦白粉病菌有性生殖与自然群体交配型检测   总被引:1,自引:0,他引:1  
 本研究根据MAT1-1-1(GenBank登录号HQ171902)和MAT1-2-1(GenBank登录号HQ171899)的部分基因序列设计和筛选其特异性引物,使用PCR方法对2012年度采自四川、陕西、云南、贵州、河南、安徽、江苏、河北、山东和甘肃10个省30个采集地点共516个小麦白粉病菌菌株的交配型基因进行检测。结果显示30个采集地点两种交配型基因都能检测到,其中28个采集地的两种交配型菌株比例(MAT1-1-1 :MAT1-2-1)符合1:1。选择具不同交配型基因的小麦白粉病菌株配对接种,16个随机杂交组合都能产生闭囊壳。对获得的闭囊壳进行子囊孢子释放,均可获得子囊孢子。该研究结果为深入研究有性生殖对小麦白粉病菌株毒性多样性的作用奠定了基础。  相似文献   

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