Garcia-Fraile Paula, Seaman Jonathan C, Karunakaran Ramakrishnan, Edwards Anne, Poole Philip S, Downie J Allan
Department of Molecular Microbiology, John Innes Centre, Norwich Research Park, Norwich, NR4 7UH UK ; Institute of Microbiology, Academy of Sciences of Czech Republic, Videnska 1083, 142 20 Prague, Czech Republic.
School of Biological Sciences, University of Reading, Reading, RG6 6AJ UK.
Plant Soil. 2015;390(1-2):251-264. doi: 10.1007/s11104-015-2389-5. Epub 2015 Jan 30.
To form nitrogen-fixing nodules on pea roots, biovar must be competitive in the rhizosphere. Our aim was to identify genes important for rhizosphere fitness.
Signature-tagged mutants were screened using microarrays to identify mutants reduced for growth in pea rhizospheres. Candidate mutants were assessed relative to controls for growth in minimal medium, growth in pea rhizospheres and for infection of peas in mixed inoculants. Mutated genes were identified by DNA sequencing and confirmed by transduction.
Of 5508 signature-tagged mutants, microarrays implicated 50 as having decreased rhizosphere fitness. Growth tests identified six mutants with rhizosphere-specific phenotypes. The mutation in one of the genes was in an arabinose catabolism operon and blocked growth on arabinose. The mutation in another gene (), encoding a predicted solute binding protein for protocatechuate and hydroxybenzoate uptake, decreased growth on protocatechuate. Both mutants were decreased for nodule infection competitiveness with mixed inoculants, but nodulated peas normally when inoculated alone. Other mutants with similar phenotypes had mutations predicted to affect secondary metabolism.
Catabolism of arabinose and protocatechuate in the pea rhizosphere is important for competitiveness of . Other genes predicted to be involved in secondary metabolism are also important.
为了在豌豆根上形成固氮根瘤,生物型必须在根际具有竞争力。我们的目的是鉴定对根际适应性重要的基因。
使用微阵列筛选签名标签突变体,以鉴定在豌豆根际中生长减少的突变体。相对于对照,评估候选突变体在基本培养基中的生长、在豌豆根际中的生长以及在混合接种物中对豌豆的感染情况。通过DNA测序鉴定突变基因,并通过转导进行确认。
在5508个签名标签突变体中,微阵列表明有50个突变体的根际适应性降低。生长测试鉴定出6个具有根际特异性表型的突变体。其中一个基因的突变位于阿拉伯糖分解代谢操纵子中,阻断了在阿拉伯糖上的生长。另一个基因()的突变,该基因编码一种预测的用于原儿茶酸和羟基苯甲酸摄取的溶质结合蛋白,降低了在原儿茶酸上的生长。这两个突变体在混合接种物中根瘤感染竞争力均降低,但单独接种时豌豆正常结瘤。其他具有相似表型的突变体的突变预计会影响次级代谢。
豌豆根际中阿拉伯糖和原儿茶酸的分解代谢对生物型的竞争力很重要。其他预计参与次级代谢的基因也很重要。