Jablonska Barbara, Ammiraju Jetty S S, Bhattarai Kishor K, Mantelin Sophie, Martinez de Ilarduya Oscar, Roberts Philip A, Kaloshian Isgouhi
Department of Nematology, University of California, Riverside, California 92521, USA.
Plant Physiol. 2007 Feb;143(2):1044-54. doi: 10.1104/pp.106.089615. Epub 2006 Dec 15.
Resistance conferred by the Mi-1 gene from Solanum peruvianum is effective and widely used for limiting root-knot nematode (Meloidogyne spp.) yield loss in tomato (Solanum lycopersicum), but the resistance is ineffective at soil temperatures above 28 degrees C. Previously, we mapped the heat-stable resistance gene Mi-9 in Solanum arcanum accession LA2157 to the short arm of chromosome 6, in a genetic interval as Mi-1 and the Cladosporium fulvum resistance gene Cf2. We developed a fine map of the Mi-9 region by resistance and marker screening of an F2 population and derived F3 families from resistant LA2157 x susceptible LA392. Mi-1 intron 1 flanking primers were designed to amplify intron 1 and fingerprint Mi-1 homologs. Using these primers, we identified seven Mi-1 homologs in the mapping parents. Cf-2 and Mi-1 homologs were mapped on chromosome 6 using a subset of the F2. Cf-2 homologs did not segregate with Mi-9 resistance, but three Mi-1 homologs (RH1, RH2, and RH4) from LA2157 and one (SH1) from LA392 colocalized to the Mi-9 region. Reverse transcriptase-polymerase chain reaction analysis indicated that six Mi-1 homologs are expressed in LA2157 roots. We targeted transcripts of Mi-1 homologs for degradation with tobacco (Nicotiana tabacum) rattle virus (TRV)-based virus-induced gene silencing using Agrobacterium infiltration with a TRV-Mi construct. In most LA2157 plants infiltrated with the TRV-Mi construct, Mi-9-mediated heat-stable root-knot nematode resistance was compromised at 32 degrees C, indicating that the heat-stable resistance is mediated by a homolog of Mi-1.
来自秘鲁番茄(Solanum peruvianum)的Mi-1基因所赋予的抗性有效,并且被广泛用于限制番茄(Solanum lycopersicum)中根结线虫(Meloidogyne spp.)造成的产量损失,但该抗性在土壤温度高于28摄氏度时无效。此前,我们将野生番茄(Solanum arcanum)种质LA2157中的热稳定抗性基因Mi-9定位到6号染色体短臂上,其遗传区间与Mi-1和抗叶霉病基因Cf2相同。我们通过对F2群体进行抗性和标记筛选,并从抗性的LA2157与感病的LA392杂交得到的F3家系中,构建了Mi-9区域的精细图谱。设计Mi-1内含子1侧翼引物来扩增内含子1并对Mi-1同源基因进行指纹分析。利用这些引物,我们在作图亲本中鉴定出7个Mi-1同源基因。使用F2的一个子集将Cf-2和Mi-1同源基因定位到6号染色体上。Cf-2同源基因与Mi-9抗性不发生分离,但来自LA2157的3个Mi-1同源基因(RH1、RH2和RH4)以及来自LA392的1个(SH1)与Mi-9区域共定位。逆转录聚合酶链反应分析表明,6个Mi-1同源基因在LA2157根中表达。我们利用基于烟草(Nicotiana tabacum)脆裂病毒(TRV)的病毒诱导基因沉默技术,通过农杆菌浸润携带TRV-Mi构建体,靶向降解Mi-1同源基因的转录本。在大多数用TRV-Mi构建体浸润的LA2157植株中,Mi-9介导的热稳定根结线虫抗性在32摄氏度时受到损害,这表明热稳定抗性是由Mi-1的一个同源基因介导的。