Buck J W, Andrews J H
University of Wisconsin, Madison, Wisconsin 53706, USA.
Appl Environ Microbiol. 1999 Feb;65(2):465-71. doi: 10.1128/AEM.65.2.465-471.1999.
The basidiomycetous yeast Rhodosporidium toruloides (anamorph, Rhodotorula glutinis) is a common phylloplane epiphyte with biocontrol potential. To understand how R. toruloides adheres to plant surfaces, we obtained nonadherent fungal mutants after chemical mutagenesis with methane-sulfonic acid ethyl ester. Sixteen attachment-minus (Att-) mutants were identified by three methods: (i) screening capsule-minus colonies for loss of adhesive ability; (ii) enrichment for mutants unable to attach to polystyrene; and (iii) selection for reduced fluorescence of fluorescein isothiocyanate-concanavalin A (Con A)-stained cells by fluorescence-activated cell sorting. None of the 16 mutants attached to polystyrene or barley leaves. The lectin Con A eliminated adhesion in all of the wild-type isolates tested. Hapten competition assays indicated that Con A bound to mannose residues on the cell surface. Adhesion of wild-type R. toruloides was transient; nonadhesive cells subsequently became adhesive, with bud development. All Att- mutants and nonattaching wild-type cells lacked polar regions that stained intensely with fluorescein isothiocyanate-Con A and India ink. Lectin, enzyme, and chemical treatments showed that the polar regions consisted of alkali-soluble materials, including mannose residues. Tunicamycin treatment reduced wild-type adhesion, indicating that the mannose residues could be associated with glycoproteins. We concluded that compounds, including mannose residues, that are localized at sites of bud development mediate adhesion of R. toruloides to both polystyrene and barley leaf surfaces.
担子菌酵母红冬孢酵母(无性型为粘红酵母)是一种常见的叶表附生菌,具有生物防治潜力。为了解红冬孢酵母如何附着于植物表面,我们用甲基磺酸乙酯进行化学诱变后获得了非附着性真菌突变体。通过三种方法鉴定出了16个附着缺陷型(Att-)突变体:(i)筛选无荚膜菌落以检测其黏附能力的丧失;(ii)富集无法附着于聚苯乙烯的突变体;(iii)通过荧光激活细胞分选选择异硫氰酸荧光素-伴刀豆球蛋白A(Con A)染色细胞荧光减弱的突变体。这16个突变体均未附着于聚苯乙烯或大麦叶片。凝集素Con A消除了所有测试的野生型菌株的黏附。半抗原竞争试验表明Con A与细胞表面的甘露糖残基结合。野生型红冬孢酵母的黏附是短暂的;非黏附细胞随后随着芽的发育而变得具有黏附性。所有Att-突变体和不附着的野生型细胞都缺乏用异硫氰酸荧光素-Con A和印度墨汁强烈染色的极性区域。凝集素、酶和化学处理表明极性区域由碱溶性物质组成,包括甘露糖残基。衣霉素处理降低了野生型的黏附,表明甘露糖残基可能与糖蛋白有关。我们得出结论,包括甘露糖残基在内的位于芽发育部位的化合物介导了红冬孢酵母对聚苯乙烯和大麦叶片表面的黏附。