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本研究以自育的26份番茄育种种质和抗南方根结线虫番茄品种‘仙客1号’及感南方根结线虫品种‘阿乃兹’为试验材料,采用病土盘栽自然发病鉴定、苗期二龄幼虫人工接种鉴定和PCR扩增及CAPS检测相结合的方法,研究供试材料对南方根结线虫的抗性水平及基因型。鉴定结果表明,供试的28份番茄材料中,高抗17份、抗病6份(包括抗病对照‘仙客1号’)、感病2份、高感3份(包括感病对照‘阿乃兹’),无免疫材料,且病土盘栽鉴定与苗期接种鉴定的结果一致。PCR扩增结果表明,27份番茄材料均能检测到一条750 bp的特异条带;TaqⅠ酶切结果显示,具有酶切位点的22份试材中有15份含纯合显性Mi-1/Mi-1基因、5份含杂合显性Mi-1/mi-1基因、2份含180 bp+250 bp+300 bp三条特异性片段,并且这22份试材对南方根结线虫表现高抗或抗,而其余5份不含显性Mi-1基因的试材则表现高感或感,证明Mi-1是具有抗南方根结线虫能力的基因,分子检测结果与病土盘栽鉴定和苗期接种鉴定的结果吻合率达92.6%;含有180 bp+250 bp+300 bp三条特异性片段的高抗材料可能含有其它抗根结线虫基因。分子检测与抗性鉴定相结合,能为创制抗南方根结线虫新种质提供一种更加省时、省力且经济有效的科学育种方法,可为番茄抗南方根结线虫育种提供理论依据和实践经验。
In this study, 26 self-bred tomato breeding germplasms and resistant to Meloidogyne incognita tomato cultivar ’Xianke 1’ and southern root-knot nematode ’Anais’ were used as experimental materials, , The second instar larvae artificial inoculation identification and PCR amplification and CAPS detection methods to study the resistance of the tested materials to southern root knot nematode level and genotype. The results showed that among 28 tomato cultivars tested, 17 were highly resistant, 6 were resistant to disease (including the resistant control ’Xianke 1’), 2 were susceptible, 3 were highly susceptible (including susceptible control ’ Arnaud ’), no immune material, and soil disease identification and seedlings inoculum identification consistent results. PCR amplification results showed that a specific 750 bp band could be detected in 27 tomato materials. The results of TaqⅠ digestion showed that 15 of the 22 samples with restriction sites contained homozygous dominant Mi-1 5 Mi-1 / mi-1 genes, two of them contained three specific fragments of 180 bp + 250 bp + 300 bp, and these 22 samples showed strong resistance to Meloidogyne incognita High resistance or resistance, while the remaining five copies of the non-dominant Mi-1 gene showed high sense of flu or test material, Mi-1 is proved to have resistance to root-knot nematode gene, the molecular test results and the soil planted The coincidence rate of identification and seedling inoculation was 92.6%. The high resistance material containing three specific fragments of 180 bp + 250 bp + 300 bp might contain other genes against root knot nematode. The combination of molecular detection and resistance identification can provide a more time-saving, labor-saving and cost-effective scientific breeding method for the development of new germplasm resistant to Meloidogyne incognita, which can provide theoretical basis and practice for tomato resistance to Meloidogyne incognita experience.