Instituto de Bioquímica Vegetal y Fotosíntesis, Consejo Superior de Investigaciones Científicas and Universidad de Sevilla, E-41092 Seville, Spain.
Plant Physiol. 2013 Apr;161(4):1984-92. doi: 10.1104/pp.112.213116. Epub 2013 Mar 5.
In cyanobacteria-plant symbioses, the symbiotic nitrogen-fixing cyanobacterium has low photosynthetic activity and is supplemented by sugars provided by the plant partner. Which sugars and cyanobacterial sugar uptake mechanism(s) are involved in the symbiosis, however, is unknown. Mutants of the symbiotically competent, facultatively heterotrophic cyanobacterium Nostoc punctiforme were constructed bearing a neomycin resistance gene cassette replacing genes in a putative sugar transport gene cluster. Results of transport activity assays using (14)C-labeled fructose and glucose and tests of heterotrophic growth with these sugars enabled the identification of an ATP-binding cassette-type transporter for fructose (Frt), a major facilitator permease for glucose (GlcP), and a porin needed for the optimal uptake of both fructose and glucose. Analysis of green fluorescent protein fluorescence in strains of N. punctiforme bearing frt::gfp fusions showed high expression in vegetative cells and akinetes, variable expression in hormogonia, and no expression in heterocysts. The symbiotic efficiency of N. punctiforme sugar transport mutants was investigated by testing their ability to infect a nonvascular plant partner, the hornwort Anthoceros punctatus. Strains that were specifically unable to transport glucose did not infect the plant. These results imply a role for GlcP in establishing symbiosis under the conditions used in this work.
在蓝细菌-植物共生体中,共生固氮蓝细菌的光合作用活性较低,由植物伙伴提供的糖来补充。然而,共生体中涉及哪些糖以及蓝细菌的糖摄取机制尚不清楚。本文构建了共生能力强、兼性异养的蓝细菌鱼腥藻属 punctiforme 的突变体,其带有一个新霉素抗性基因盒,取代了一个假定的糖转运基因簇中的基因。使用(14)C 标记的果糖和葡萄糖进行的转运活性测定结果以及这些糖的异养生长测试,确定了一个用于果糖(Frt)的 ATP 结合盒型转运蛋白、一个用于葡萄糖的主要易化渗透酶(GlcP)以及一个对果糖和葡萄糖最佳摄取都需要的孔蛋白。对带有 frt::gfp 融合基因的鱼腥藻属 punctiforme 菌株的绿色荧光蛋白荧光分析表明,在营养细胞和类似厚垣孢子的细胞中高表达,在游动孢子中可变表达,在异形胞中无表达。通过测试鱼腥藻属 punctiforme 糖转运突变体感染非维管束植物伙伴角苔 Anthoceros punctatus 的能力,研究了其共生效率。不能特异性运输葡萄糖的菌株不能感染植物。这些结果表明,在本工作中使用的条件下,GlcP 在建立共生关系中起作用。