单孢分离菌株的异核不对称特性揭示梯棱羊肚菌为假同宗结合真菌

本文以基因组数据显示只具有MAT1-1 idiomorph的单孢菌株YPL6-3和只具有MAT1-2 idiomorph的单孢菌株YPL6-1为材料,研究它们子代的子囊果、单孢菌株群体和同一子囊中8个单孢菌株的交配型分布情况。YPL6-3和YPL6-1菌株分别隔离栽培和互补混栽均能形成正常的子囊果,其子囊果菌柄交配型分布与亲本菌株有关。PCR扩增检测235株子代单孢菌株的交配基因出现有趣现象：一些菌株的MAT1-1-1基因电泳条带强,而MAT1-2-1基因的条带弱;另一些菌株则MAT1-1-1基因条带弱,而MAT1-2-1基因条带强。同时也有两基因条带都强或者一基因条带强,另一基因无条带的菌株。从3个子囊果中共挑取了10个子囊,并对每个子囊中的单孢进行独立分离,PCR扩增并电泳检测交配基因时也出现了上述相同的情况。若MAT1-1-1强,则MAT1-2-1弱或无,这样的菌株在同一子囊中不会超出4个,反之亦然。利用长片段PCR扩增YPL6-1和YPL6-3菌株的全长MAT idiomorph,利用Nanopore测序技术对扩增子进行单分子实时测序,2次重复实验的序列比对发现：菌株YPL6-1中存在99.63%和99.81%的MAT1-2 idiomorph分子及0.37%和0.19%的MAT1-1 idiomorph分子;菌株YPL6-3中存在99.45%和99.74%的MAT1-1 idiomorph 分子及0.55%和0.26%的MAT1-2 idiomorph分子。从而证实,这2个基因组测序和PCR扩增都只能检测到一种交配型的菌株,其实是异核菌株,只是2种交配型核的数量占比存在较大偏离。根据上述现象推断,梯棱羊肚菌的子囊孢子都是异核的,萌发后形成的单孢菌株具有异核不对称特点,从而推测梯棱羊肚菌是一种特殊的假同宗子囊菌,同时也揭示了梯棱羊肚菌单孢出菇的真相。     

Heterothallism in Saccharomyces cerevisiae isolates from nature: effect of HO locus on the mode of reproduction

Understanding the evolution of sex and recombination, key factors in the evolution of life, is a major challenge in biology. Studies of reproduction strategies of natural populations are important to complement the theoretical and experimental models. Fungi with both sexual and asexual life cycles are an interesting system for understanding the evolution of sex. In a study of natural populations of yeast Saccharomyces cerevisiae , we found that the isolates are heterothallic, meaning their mating type is stable, while the general belief is that natural S. cerevisiae strains are homothallic (can undergo mating-type switching). Mating-type switching is a gene-conversion process initiated by a site-specific endonuclease HO; this process can be followed by mother–daughter mating. Heterothallic yeast can mate with unrelated haploids (amphimixis), or undergo mating between spores from the same tetrad (intratetrad mating, or automixis), but cannot undergo mother–daughter mating as homothallic yeasts can. Sequence analysis of HO gene in a panel of natural S. cerevisiae isolates revealed multiple mutations. Good correspondence was found in the comparison of population structure characterized using 19 microsatellite markers spread over eight chromosomes and the HO sequence. Experiments that tested whether the mating-type switching pathway upstream and downstream of HO is functional, together with the detected HO mutations, strongly suggest that loss of function of HO is the cause of heterothallism. Furthermore, our results support the hypothesis that clonal reproduction and intratetrad mating may predominate in natural yeast populations, while mother–daughter mating might not be as significant as was considered.     

Experimental sexual selection in Chlamydomonas

Sexual and asexual lines of the unicellular chlorophyte Chlamydomonas reinhardtii were propagated for about 100 sexual cycles and 1000 vegetative cycles in contrasted environments, liquid and solid growth media, in order to generate divergent natural and sexual selection. Sexual lines were transferred by many zygotes or by a single zygote in each sexual generation. By the end of the experiment zygote production was in the order sexual mass-transfer>sexual single-zygote>asexual>ancestor. The direct response to sexual selection was large, with zygote production increasing by about two orders of magnitude, mainly because mating had become spontaneous instead of being invoked by nitrogen starvation. Asexual lines became sexually sterilized by the fixation of a single mating type. Sexual selection caused a radical shift in the gender system, with homothallism spreading to high frequency in all sexual lines of this normally heterothallic species. This may have been caused by the transposition of a mating-type gene to an autosome. No substantial degree of environment-specific mating evolved, however, and thus no sexual isolation indicative of incipient speciation. It is possible that selection experiments of this kind are unlikely to induce sexual isolation because mating-type genes evolve in a saltatory fashion.     

黄伞的交配型性状研究

以两个黄伞子实体为材料, 经担孢子弹射, 稀释分离获得两组Ⅰ和Ⅱ分别为27和104株单核体。通过采用三轮杂交系统研究,确定黄伞为四极性异宗配合的担子菌;实验还发现单核体不同类型在群体中的比例显示出明显的偏向性分布。     

梯棱羊肚菌单孢及杂交群体的栽培出菇试验和极性分析

羊肚菌的极性和单孢出菇至今未有确切的实证,这限制了对羊肚菌生活史的深入理解和遗传育种工作的开展。本研究选用梯棱羊肚菌(Morchella importuna)栽培菌株(F0),采取显微操作从其正常出菇的子囊果获得F1单孢群体;随机选取其中的7个F1单孢菌株和10对单孢杂交(F1两两混合,记为F2)进行栽培试验,记录和分析了栽培出菇时间、采收期、产量和其他生产性状。此外,分析了F0亲本菌株、F1单孢群体、F2子囊果以及2个F1单孢菌株出菇的F2子囊果的F3单孢群体的交配型基因型。结果表明:除了F1单孢菌株MM32未出菇以外,其他供试菌株均正常出菇。各菌株的出菇潜能不同,3个F1单孢菌株的平均单产高于出发菌株,其中MM34菌株平均单产最高,达1.01 kg/m^2,是F0亲本菌株(0.68 kg/m^2)的1.49倍; 10个单孢杂交的F2平均单产在0.21～0.97 kg/m^2之间。F0亲本菌株和所有的F2出菇子囊果中均含有2种交配型基因,F1和F3单孢群体仅含有2种交配型基因中的1种。虽然F1单孢群体大部分菌株均正常出菇,然而F2子囊果和F3单孢群体的交配型基因型分析结果表明,单孢菌株在栽培生产中自然引入了具有相反交配型基因的细胞核,这种自然引入可能是由无性孢子传播引起。出菇试验和交配型基因分析确证梯棱羊肚菌为异宗结合真菌。本研究有利于深入理解羊肚菌生活史,促进羊肚菌育种工作的开展。     

Fruiting studies in species ofCapronia (Herpotrichiellaceae)

A recently described protocol for thein vitro production of ascomata was employed to determine the sexual incompatibility systems of five species ofCapronia. The formation of mature ascomata in isolates derived from single ascospores demonstrated thatC. epimyces, C. mansonii, andC. munkii n. sp. are homothallic. In contrast, fertile ascomata were observed only in mass-ascospore isolates and pairwise crosses between specific single-ascospore isolates inC. dactylotricha n. sp. andC. moravica. TheExophiala anamorphs ofC. dactylotricha andC. munkii are described and aPhialophora-like synanamorph is reported for the former species. Germinating ascospores ofC. munkii formed conidiogenous cells directly, while the ascospores of the remaining species germinated to produce germ tubes and hyphae. The application of the terms microcyclic conidiation to secondary conidium production and sclerotial body and stroma to the multicellular structures produced by species ofCapronia andExophiala are discussed.     

Wirtelschnallen bei homo- und heterothallischenCoprinus-Arten (Agaricales)

Within agaric fungi whorled (multiple) clamps have been described so far only for the homothallicCoprinus myceliocephalus Lange. The new speciesCoprinus verticillatus Schulz-Weddigen (ined.) develops fertile mycelia which form septae without or with 1–5 clamps. Stronger hyphae tend to branch dichotomously. Most cells of the vegetative system contain many nuclei. Though single spore cultures remain sterile, they can develop multiple clamps, too. Moreover, very young mycelia produce a special type of oidiophores. According to its mating interactions a bipolar pattern characterizes the heterothallicC. verticillatus.

     

Tetrasporangia in Asparagopsis armata

The first record for tetrasporangia in Asparagopsis armata Harv. (Falkenbergia rufolanosa (Harv.) Schm.) is given for the British Isles.     

EVIDENCE FOR SEXUAL REPRODUCTION IN THE PRIMITIVE GREEN ALGA NEPHROSELMIS OLIVACEA (PRASINOPHYCEAE)1

Evidence for a specialized sexual process in Nephroselmis olivacea Stein is presented. This alga is a member of the Prasinophyceae, which is regarded by some as the most primitive Class of green plants. N. olivacea has a heterothallic type of mating system. Plus and minus gametes were morphologically similar but showed different behaviors during the mating process. The minus gamete settled to the substratum, attaching by its ventral side. The plus gamete attached to the dorsal side of the minus gamete by the region near the flagellar bases of the plus gamete. The mature zygotes were spherical and strongly adhered to the substratum. After zygote germination, two biflagellate daughter cells, each with two pyrenoids were liberated. These cells divided, resulting in four vegetative cells, each with a single pyrenoid.