ISOZYME ANALYSIS OF GENETIC DIVERSITY AMONG ISOLATES OF RHIZOCTONIA SOLANI ANASTOMOSIS GROUP 1-Ⅰ A(AG1-Ⅰ A)

利用系统聚类分析方法,通过对丝核菌菌丝融合一群Ｒｈｉｚｏｃｔｏｎｉａｓｏｌａｎｉ（ＡＧ１）的２３个菌株在七个同功酶电泳所显示的５０个表现型进行了遗传分析。结果显示,２３个丝核菌菌株可聚类为三大类,他们分别代表了丝核菌菌丝融合一群中的三个亚群ＡＧⅠＡ,ＡＧⅠＢ,ＡＧⅠＣ;其中第一聚类又可进一步分为四个亚类。同功酶的分析证实了丝核菌菌丝融合１群中的三个亚群的分类概念是具有遗传背景的,并可作为丝核菌菌丝融合群之间及群内分类的重要手段。     

利用同功酶分析丝核菌菌丝融合1群1A亚群：（RhizoctoniasolaniAG1—1 …

利用系统聚类分析方法，通过对丝核菌菌丝融合一群Ｒｈｉｚｏｃｔｏｎｉｃｓｏｌａｎｉ（ＡＧ－１）的２３个菌株在七个同功酶电泳所显示的５０个表现型进行了遗传分析。     

三种蜘蛛丝蛋白组成分析

本文应用高压液相色谱（ＨＰＬＣ）法分析了岳麓山的大腹园蛛Ａｒａｎｅｕｓ ｖｅｎｔｒｉｃｏｓｕｓ（Ｃ．Ｋｏｃｈ,１８７８）,机敏漏头蛛Ａｇｅｌｅｎａ ｄｉｆｆｉｃｌｉｓ （Ｆｏｘ,１９３７）,白额巨蟹蛛Ｈｅｔｅｒｏｐｏｄａ ｖｅｎａｔｏｒｉａ （Ｌｉｎｎａｅｕｓ,１７５７）的丝蛋白的氨基酸组成,以ＳＤＳ－ＰＡＧＥ法测定了大腹园蛛不同丝腺体的未成丝的可溶性丝蛋白的分子量。实验结果表明蛛丝蛋白中占优势的     

丛枝菌根真菌对植物根尖分生区和根冠细胞的侵染*

一般说来,从枝菌根（ＡＭ）真菌大多数是从植物根系根毛区（成熟区）侵入和扩展的,在显微镜下往往看不到根尖分生区和根冠表皮细胞被ＡＭ真菌侵染的特征。这就很容易给人们造成一种假象,似乎ＡＭ真菌不能侵染根尖分生区和根冠表皮细胞,即它们对ＡＭ真菌是免疫的。然而笔者多次于显微镜下看到ＡＭ真菌侵染根尖分生区和根冠表皮细胞,并形成典型的泡囊、丛枝、菌丝等结构。这一现象导致作者在温室盆栽和大田条件下研究了玫瑰红巨孢囊霉（ Ｇｉｇａｓｐｏｒａ　ｒｏｓｅａ Ｎｉｃｏｌ　＆ Ｓｃｈｅｎｃｋ）、珠状巨孢囊霉（Ｇｉｇａｓｐｏｒａ　ｍａｒｇａｒｉｔａ　Ｂｅｃｋｅｒ　＆　Ｈａｌｌ）、根内球囊霉（Ｇｌｏｍｕｓ　ｏｍｔｒａｒａｄｉｃｅｓ　ｓｃｈｅｎｃｋ　＆　Ｓｍｉｔｈ、摩西球囊霉（Ｇｌｏｍｕｓ ｍｏｓｓｅａｅ （Ｎｉｃｏｌ　＆ Ｇｅｒｄ．） Ｇｅｒｄｅｍａｎｎ　＆ Ｔｒａｐｐｅ）、地表球囊霉（ Ｇｌｏｍｕｓ ｖｅｒｓｉｆｏｒｍｅ（ Ｋａｒｓｔｅｎ）Ｂｅｒｃｈ）和弯丝硬囊霉（ Ｓｃｌｅｒｏｃｙｓｔｉｓ　ｓｉｎｕｏｓａ　Ｇｅｒｄｅｍａｎｎ　＆ Ｂａｋｈｉ）对棉花（Ｇｏｓｓｙｐｉｕｍ　ｈｉｒｓｕｔｕｍ　Ｌ．）、烟草（Ｎｉｃｏｔｉａｎａ　 ｔａｂａｃｕｍ Ｌ．）和白     

识别未衍生化的13—羟化GAs及其葡萄糖苷的单克隆抗体

抗ＧＡ３ 及其葡萄糖苷的ＭＡＢ１０单克隆抗体源于以ＧＡ３ 中的３ 位羟基（３－ＯＨ）为偶联位点,人血清白蛋白（ＨＳＡ）为载体合成的ＧＡ３－３－ＨＳＡ 免疫原． 该抗体对１３－羟化ＧＡｓ（１３－ＯＨＧＡｓ、ＧＡ１、ＧＡ３、ＧＡ５ 等）和ＧＡ３ 葡萄糖苷具有高亲和力． ７ 位羧基的甲酯化可显著降低ＭＡＢ１０ 对１３－ＯＨ ＧＡｓ的亲和力,而３－ＯＨ 的糖苷化却未降低其亲和力． 用该抗体建立的两种分别用于ＧＡ３ 及其葡萄糖苷测定的酶联免疫吸附法（ＥＬＩＳＡ）,其检测线性范围均为０．２～２０ ｐｍ ｏｌ． 借助这两种ＥＬＩＳＡｓ,研究了羊蹄（Ｒｕｍ ｅｘ ｊａｐｏｎｉｃｕｓ）叶片中ＧＡ３ 及其类似ＧＡｓ和葡萄糖苷的动态变化．结果表明,叶片衰老与游离态ＧＡｓ的糖苷化有关;而６－ＢＡ 延缓衰老则可能与其减缓ＧＡｓ的糖苷化有关     

中国附丝壳属研究

本文报道了附丝壳属Ａｐｐｅｎｄｉｃｕｌｅｌｌａ ｖｏｎ Ｈｏｅｈｎｅｌ两个新种。它们是：蕈树附丝壳Ａｐｐｅｎｄｉｃｕｌｅｌｌａ ａｌｔｉｎｇｉａｅ Ｙ．Ｘ．Ｈｕ ｅｔ Ｂ．Ｓｏｎｇ、武夷附丝壳Ａｐｐｅｎｄｉｃｕｌｅｌｌａ ｗｕｙｉｅｎｓｉｓ Ｙ．Ｘ．Ｈｕ ｅｔ Ｂ．Ｓｏｎｇ。新种附有拉丁文、中文描述和显微结构图。模式标本藏于广东省微生物研究所真菌标本室（ＨＭＩＧＤ）。     

滇南及滇东南胶孔菌复合群的分类地理研究

报道了我国滇南及滇东南热带、亚热带胶孔菌复合群众路线属６种,其中瘤丝牛肝菌Ｆｉｌｏｂｏｌｅｔｕｓ ｖｅｒｒｕｃｕｌｏｓｕｓ Ｐ．Ｇ．Ｌｉｕ和滇丝牛肝菌Ｆ．ｙｕｎｎａｎｅｎｓｉｓ Ｐ．Ｇ．Ｌｉｕ是新种,紫兰小菇Ｍｙｃｅｎａ ｖｉｏｌａｃｅｌｌａ（Ｓｐｅｑ．）Ｓｉｎｇ。是我国新纪录种;新种附有拉丁文描术和插图,新纪录和附有形态解剖图。本文所引证标本均存放于中国科学院昆明植物研究所隐花植物标本馆（ＨＫＡ     

中国附丝壳属研究III.

本文报道了中国的附丝壳属Ａｐｐｅｎｄｉｃｕｌｅｌｌａ二个新种,八角附丝壳ＡｐｐｅｎｄｉｃｕｌｅｌｌａｉｌｌｉｃｉｉＢ．Ｓｏｎｇ和新木姜子附丝壳ＡｐｐｅｎｄｉｃｕｌｅｌｌａｎｅｏｌｉｔｓｅａｅＢ．Ｓｏｎｇ新种附有拉丁文,英文描述和显微构图,模式标本保存于广东省微生物研究所标本室（ＨＭＩＧＤ）。     

管藻目绿藻叶绿素蛋白复合物特性及比较研究

采用温和的ＰＡＧＥ法从管藻目刺松藻（Ｃｏｄｉｕｍ ｆｒａｇｉｌｅ （Ｓｕｒ．）Ｈａｒｉｏｔ）和假根羽藻（Ｂｒｙｏｐｓｉｓ ｃｏｒｔｉｃｕｌａｎｓ Ｓｅｔｃｈ．）,丝藻目绿藻软丝藻（Ｕｌｏｔｈｒｉｘ ｆｌａｃｃａ （Ｄｉｌｌｗ．）Ｔｈｕｒ．）,及菠菜（Ｓｐｉｎａｃｉａ ｌｏｅｒａｃｅａ Ｍｉｌｌ．）中分别得到１１、１１、７和９种色素蛋白复合物,对复合物的多种特性,包括分子量、Ｃｈｌ ａ／ｂ比值、叶绿     

牡丹冬季到内催花过程中内源激素含量的变化

牡丹品种朱砂垒（Ｐａｅｏｎｉａ ｓｕｆｆｒｕｔｉｃｏｓａ Ａｎｄｒ．ｃｖ．Ｚｈｕｓｈａｌｅｉ）在冬季室内催花过程７种内源激素含量变化不同。玉米素核苷（Ｚ＋ＡＲ）、生长素（ＩＡＡ）和赤霉素（ＧＡ３）的含量在花生长发 处于较高水平;而脱落酸（ＡＢ）、异戊烯基腺苷（ＩＰ＋ＩＰＡ）、二氢玉米素核苷（ＤＨＺ＋ＤＨＺＲ）、赤霉素（ＧＡ４）的含量低于上述３种内源激素。激素平衡方面,ＧＡｓ／ＡＢＡ、ＣＴＫｓ／ＡＢ     

立枯丝核菌可溶性蛋白图谱研究 Ⅱ.用等电聚焦电泳比较各融合群及亚群

利用薄层(0.5mm)聚丙烯酰胺凝胶等电聚焦电泳对于pH3.5-10.0范围内立枯丝核菌(Rhizoctonia solani)AG-1～AG-5各融合群及亚群共45个参试菌株的菌体可溶性蛋白进行了比较分析。参试菌株分别来自于日本、美国及国内鉴定的菌株。电泳结果表明:不同融合群或亚群的蛋白质图谱表现显著差异。AG-3、AG-4、A G-5各融合群和AG-1IA、IB,IC、AG-2-1、AG-2-2各亚群分别表现出各自的特征性图谱。以上结论与前篇(刘力、葛起新,1988)报道中的基本相符,且更为明确。针对试验结果,就可溶性蛋白等电聚焦电泳图谱与培养性状类型的比较以及不同菌培养时间对电泳结果的影响进行了分析讨论。     

中国东北地区玉米纹枯病菌的融合群鉴定

自东北三省采集玉米纹枯病标本300余份,分离获得286个丝核菌菌株。融合群测定及5.8S rDNA-ITS区序列分析结果表明,这些菌株分别属于多核丝核菌的AG1-IA、AG1-IB、AG1-IC、AG4-HG-I、AG4-HG-III、AG-5、WAG-Z群及双核丝核菌的AG-Ba群。其中AG1-IA是优势致病群,占分离菌株总数的38.46%,其次是WAG-Z和AG-5群,分别占26.92%及24.83%。AG4-HG-III群菌株是国内首次从罹病玉米植株上分离得到。自各融合群中选取代表性的菌株进行5.8     

Characterization of Xanthomonas oryzae pv. oryzae recX, a gene that is required for high-level expression of recA

A universally primed (UP)-PCR cross hybridization assay was developed for rapid identification of isolates of Rhizoctonia solani into the correct anastomosis group (AG). Twenty-one AG tester isolates belonging to 11 AGs of R. solani were amplified with a single UP primer which generated multiple PCR fragments for each isolate. The amplified products were spotted onto a filter, immobilized and used for cross hybridization against amplification products from the different isolates. Isolates within AG subgroups cross hybridize strongly, whereas between different AGs little or no cross hybridization occurs. Sixteen Rhizoctonia isolates from diseased sugar beets and potatoes were identified using the assay. The results were supported by restriction fragment length polymorphism analysis of the ITS1-5.8S-ITS2 region of the nuclear encoded ribosomal DNA. Through standardization and use of quick non-radioactive labeling techniques, the UP-PCR cross hybridization assay has potential for routine use by modern DNA chip technology.     

First Report of Rhizoctonia solani AG‐7 on Cotton in Egypt

Eighty‐two isolates of Rhizoctonia solani were recorded from roots of naturally‐infected seedlings of the Egyptian cotton (Gossypium barbadense L.). Anastomosis groups (AGs) of the isolates were determined by using 13 different AGs testers. Three (3.7%) of the isolates were identified as R. solani AG7, while the remaining isolates were belonging to the AG 2‐1, AG4 and AG5. The identification of the three isolates was based on the frequency of the C2 reaction with the AG7 tester isolate. No fusion was observed between AG7 and isolates representing the other 13 AGs. Colonies of AG7 isolates grown on potato dextrose agar (PDA), malt yeast agar (MYA) and melt peptone agar (MPA) were brown to dark brown with aerial mycelium and sclerotia. The isolates had pitted sclerotial clusters and brownish exudates after 21 days of culturing on PDA, but without clear zonation. Pathogenicity test under greenhouse conditions revealed that AG7 caused the common symptoms of damping–off, which included seed rot, lesions on the hypocotyls and root rot.     

黄淮海地区夏玉米纹枯病菌的融合群鉴定

从黄淮海地区采集玉米纹枯病标样250余份,分离得到176个丝核菌菌株。融合群测定及5.8SrDNA-ITS区序列分析结果表明,这些菌株分别属于多核丝核菌的AG1-IA、AG1-IB、AG4-HG-I、AG-5、WAG-Z融合群及双核丝核菌的AG-A、AG-Ba融合群。其中AG1-IA是优势融合群,占分离菌株总数的64.20%,其次是AG-Ba,占12.50%,再依次分别是WAG-Z(10.23%)、AGI-IB(5.11%)、AG-4-HG-I(3.98%)、AG-5(2.27%)和AG-A(1.70%)。其中AG-Ba融合群是国内首次在玉米上分离得到。从各融合群中选取代表性的菌株进行5.8S rDNA-ITS区序列分析结果表明,隶属不同融合群或亚群的菌株其5.8S rDNA-ITS区序列存在较大的差异,而相同融合群(或亚群)不同菌株之间其序列的一致性可高达97%-100%。     

Typing of anastomosis groups of Rhizoctonia solani by restriction analysis of ribosomal DNA

A method based on restriction analysis of polymerase chain reaction (PCR)-amplified ribosomal DNA was developed for the rapid characterization of large populations of Rhizoctonia solani at the anastomosis group (AG) level. The restriction maps of the internal transcribed spacers (ITS) sequences were compared for 219 isolates of R. solani belonging to AG-1 to AG-12 and AG-BI, representing diverse geographic and host range origins. Four discriminant restriction enzymes (MseI, AvaII, HincII, and MunI) resolved 40 restriction fragment length polymorphism (RFLP) types among the 219 ITS sequences of R. solani. Each RFLP type could be assigned to a single AG except for two RFLP types, which were common to two AG. A fifth enzyme allowed the discrimination of AG-6 and AG-12. In addition, the combination of four enzymes allowed the discrimination of subsets within AG-1, AG-2, AG-3, and AG-4. The efficiency of the typing method was confirmed by analyzing PCR-amplified ITS sequences of 30 reference strains. Furthermore, the PCR-RFLP method was used to characterize at the AG level 307 isolates of R. solani originating from ten sugar beet fields exhibiting patches of diseased plants in France. The PCR-based procedure described in this paper provides a rapid method for AG typing in R. solani.     

Cropping Systems and Cultural Practices Determine the Rhizoctonia Anastomosis Groups Associated with Brassica spp. in Vietnam

Ninety seven Rhizoctonia isolates were collected from different Brassica species with typical Rhizoctonia symptoms in different provinces of Vietnam. The isolates were identified using staining of nuclei and sequencing of the rDNA-ITS barcoding gene. The majority of the isolates were multinucleate R. solani and four isolates were binucleate Rhizoctonia belonging to anastomosis groups (AGs) AG-A and a new subgroup of A-F that we introduce here as AG-Fc on the basis of differences in rDNA-ITS sequence. The most prevalent multinucleate AG was AG 1-IA (45.4% of isolates), followed by AG 1-ID (17.5%), AG 1-IB (13.4%), AG 4-HGI (12.4%), AG 2-2 (5.2%), AG 7 (1.0%) and an unknown AG related to AG 1-IA and AG 1-IE that we introduce here as AG 1-IG (1.0%) on the basis of differences in rDNA-ITS sequence. AG 1-IA and AG 1-ID have not been reported before on Brassica spp. Pathogenicity tests revealed that isolates from all AGs, except AG-A, induced symptoms on detached leaves of several cabbage species. In in vitro tests on white cabbage and Chinese cabbage, both hosts were severely infected by AG 1-IB, AG 2-2, AG 4-HGI, AG 1-IG and AG-Fc isolates, while under greenhouse conditions, only AG 4-HGI, AG 2-2 and AG-Fc isolates could cause severe disease symptoms. The occurrence of the different AGs seems to be correlated with the cropping systems and cultural practices in different sampling areas suggesting that agricultural practices determine the AGs associated with Brassica plants in Vietnam.     

Evolutionary Diversification Indicated by Compensatory Base Changes in ITS2 Secondary Structures in a Complex Fungal Species, <Emphasis Type="Italic">Rhizoctonia solani</Emphasis>

The rRNA cistron (18S–ITS1–5.8S–ITS2–28S) is used widely for phylogenetic analyses. Recent studies show that compensatory base changes (CBC) in the secondary structure of ITS2 correlate with genetic incompatibility between organisms. Rhizoctonia solani consists of genetically incompatible strain groups (anastomosis groups, AG) distinguished by lack of anastomosis between hyphae of strains. Phylogenetic analysis of internal transcribed spacer (ITS) sequences shows a strong correlation with AG determination. In this study, ITS sequences were reannotated according to the flanking 5.8S and 28S regions which interact during ribogenesis. One or two CBCs were detected between the ITS2 secondary structure of AG-3 potato strains as compared to AG-3 tobacco strains, and between these two strains and all other AGs. When a binucleate Rhizoctonia species related to Ceratobasidiaceae was compared to the AGs of R. solani, which were multinucleate (3–21 nuclei per cell), 1–3 CBCs were detected. The CBCs in potato strains of AG-3 distinguish them from AG-3 tobacco strains and other AGs yielding further evidence that the potato strains of AG-3 originally described as R. solani are a species distinct from other AGs. The ITS1–5.8S–ITS2 sequences were analyzed by direct sequencing of PCR products from 497 strains of AG-3 isolated from potato. The same 10 and 4 positions in ITS1 and ITS2, respectively, contained variability in 425 strains (86%). Nine different unambiguous ITS sequences (haplotypes) could be detected in a single strain by sequencing cloned PCR products indicating that concerted evolution had not homogenized the rRNA cistrons in many AG-3 strains. Importantly, the sequence variability did not affect the secondary structure of ITS2 and CBCs in AG-3. Electronic supplementary material  The online version of this article (doi:) contains supplementary material, which is available to authorized users.     

Trials of direct detection and identification of <Emphasis Type="Italic">Rhizoctonia solani</Emphasis> AG 1 and AG 2 subgroups using specifically primed PCR analysis

Specifically primed polymerase chain reaction (PCR) analysis was used for direct detection and identification of Rhizoctonia solani isolates belonging to AG 1 subgroups (IA, IB, and IC) and AG 2 subgroups (2-1 and 2-2). A rapid DNA extraction method with a solution of sodium hydroxide was conducted to extract PCR templates. PCR-specific primer sets for each group were designed from sequences in the regions of the 28S ribosomal DNA of this fungus. The results of specifically primed PCR analysis showed that AG 1-IA, AG 1-IB, AG 1-IC, AG 2-1, and AG 2-2 primers sets contributed detection from the same AG isolates and could escape detection from different AG isolates at a high level of frequency. In this experiment, we suggested that our synthesized primer sets might provide a method for the direct detection and identification of AGs of R. solani. Received: June 28, 2001 / Accepted: November 14, 2001