| 摘要: |
| 为选育抗根结线虫的李属砧木,以新疆野生樱桃李(Prunus sogdiana)扦插苗为试验材料,对其接种南方根结线虫(Meloidogyne incognita),从中筛选出12株抗病植株。根据已知的抗根结线虫基因的NBS-LRR保守结构域设计简并引物,并以抗病植株的DNA和cDNA为模板对抗根结线虫同源基因序列进行扩增。获得6个抗根结线虫同源基因片段,依次命名为psoRPM1、psoRPM2、psoRPM3、psoRPM4、psoRPM5和psoRPM6,GenBank登录号为:HM593974、HM593975、HM593970、HM593971、HM593972和HM593973。其中前4个基因片段与桃、甜樱桃、酸樱桃以及樱花中的抗病基因的同源性高达90%以上。基因序列分析表明,这6个基因片段均具有典型的NBS保守结构域。根据其氨基酸序列的特点,推断这6个基因片段均属于non TIR-NBS-LRR型抗病基因。RT-PCR表达分析结果显示,psoRPM1和psoRPM6在接种南方根结线虫12和24 h后表达量显著增加,预示着这2个基因片段可能参与了樱桃李对根结线虫的识别和防御过程。本研究在樱桃李中克隆出抗根结线虫相关的NBS-LRR型基因片段,为进一步克隆樱桃李中的抗根结线虫基因全长序列及转基因育种奠定了基础。 |
| 关键词: 樱桃李 根结线虫 抗病植株 基因克隆 NBS-LRR 表达分析 |
| DOI：10.11841/j.issn.1007-4333.2010.05.013 |
| 投稿时间：2010-03-05 |
| 基金项目:国家自然科学基金资助项目 (30971981); 新疆维吾尔自治区科技计划项目 (200931101-2)第一作者:肖芳权,硕士研究生,E-mail:xfq0529@163.com通讯作者:胡建芳,副教授,主要从事果树生理与分子生物学研究,E-mail:hujf@cau.edu.cn |
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| Cloning and expression analysis of fragments related toroot-knot nematode resistance from wildMyrobalan plum (Prunus sogdiana) |
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XIAO Fang-quan1, HU Jian-fang1, XU Zheng2, LIAO Kang3, LI Tian-zhong1
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| (1.College of Agronomy and Biotechnology,China Agricultural University,Beijing 100193,China;2.YiLi Kazak Autonomous Prefecture Academy of Forestry,Yining 835000,China;3.College of Horticulture,Xinjiang Agricultural University,Wulumuqi 830052,China) |
| Abstract: |
| In order to screen the resistant rootstocks for Prunus,the resistance to root-knot nematode was evaluated with inoculating root-knot nematode Meloidogyne incognita,and then identifying the resistant plants.The resistant gene analogs from DNA and cDNA of Myrobalan plum were amplified by using degenerated primers based on the conserved regions of Nucleotide Binding Site (NBS)-Leucine Rich Repeat (LRR) domain from plant resistance genes reported.Six resistance gene analogs were obtained and named psoRPM1,psoRPM2,psoRPM3,psoRPM4,psoRPM5 and psoRPM6(GenBank accession number: HM593974,HM593975,HM593970,HM593971,HM593972,HM593973),respectively.Putative amino acid sequences deduced from the first four RGAs shared above 90% identity with the resistance genes from peach,sour cherry,sweet cherry and cherry blossom.All the six RGAs contained the conserved regions of Nucleotide Binding Site (NBS) and belonged to the non TIR-NBS-LRR class.The RT-PCR showed that the expression of the psoRPM1 and the psoRPM6 was significantly increased at 12 and 24 h after the inoculation of M.incognita,indicating the two genes might play important roles in the resistance to the root-knot nematodes.The first cloning of the RGAs from the Myrobalan plum laid a promising foundation for cloning resistance genes from Myrobalan plum and molecular breeding. |
| Key words: Myrobalan plum root-knot nematodes gene clone resistant plants NBS-LRR expression analysis |
