摘要
大豆孢囊线虫 (HeteroderaglycinesIchinohe ;SoybeanCystNematode ,SCN)是一种土传的专性内寄生线虫。SCN的二龄幼虫侵入到大豆幼嫩的根组织中 ,导致大豆根内的细胞变形并与之形成“合胞体”。合胞体在形态上和生理上的变化是SCN直接诱导大豆基因表达的结果。本研究以高抗SCN的灰布支黑豆为材料 ,用大豆孢囊线虫二龄幼虫直接接种大豆的根系 ,应用DDRT -PCR技术及RDB (Reversedot-blotting)杂交鉴定 ,获得 6个阳性cDNA克隆 ,分别是SCN侵染后 5天的A32克隆 (GenBank登录号为BI173978) ;侵染后 10天的B12克隆 (GenBank登录号为BI173979)、B71克隆 (GenBank登录号为BI173980 ) ;侵染后 15天的C11克隆 (GenBank登录号为BI173981)、CP12 (GenBank登录号为BI173982 )克隆和CP32克隆 (GenBank登录号为BI173983)。序列的同源比较表明 ,6个cDNA均与Shoemaker构建的大豆基因表达库中的cDNA序列有非常高的同源性 ,证明这些cDNA是大豆基因表达的产物。其中A32克隆的序列与控制拟南芥下胚轴生长的MYB转录因子、营养元素缺失诱导的番茄根的表达文库中的一个cDNA及番茄抗假单胞杆菌表达文库中的一个cDNA有较高的同源性。
Soybean cyst nematode (SCN), Heterodera glycines Ichinohe, is one kind of soil sedentary endoparasitic nematodes and one of the most important parasites of soybean. The second-stage juvenile (J2) of SCN penetrates soybean roots and causes the formation of specialized feeding cells which are usually called syncytium in the roots' vascular system. The morphological and physiological changes of syncytium are considered to be the results of soybean gene expression induced by SCN directly. However, the mechanism of those changes are still not very clear, identifying and characterizing resistant gene is very important to make clear the interaction between SCN and soybean. ZDD2315, a local germplasm of Shanxi province, is strong resistance to SCN race 4 and was used in this research. 2-day-old soybean seedlings were inoculated by J2 of SCN race 4. mRNA Differential display was used to detect host gene expression changes during interaction between soybean and SCN. Thirty-six cDNA clones corresponding to mRNAs with different abundances in SCN-infected (19 cDNAs) and uninfected (17 cDNAs) roots were identified. Six of them were recovered as plasmid clones that showed hybridization intensity differences in reverse dot-blotting assay. They are A32 clone (BI173978) from soybean roots five days after inoculation, B12 clone (BI173979) and B71 clone(BI173980) from soybean roots ten days after inoculation, C11 clone (BI173981), CP12 clone (BI173982) and CP32 clone (BI173983)from soybean roots fifteen days after inoculation. The sequence of above six clones have been registered in GenBank. Accession numbers are showed in the brackets. The sequences of six clones were subjected to database comparisons with the aid of BLAST algorithm. All of them showed strong similarity to cDNAs which were in soybean EST library constructed by Shoemaker. Especially, A32 clone showed strong similarity to EST cDNA for coding MYB of Arabidopsis late elongated hypocityl protein, one of cDNA from the library induced by tomato root nutrition deficiency
出处
《分子植物育种》
CAS
CSCD
2003年第2期193-200,共8页
Molecular Plant Breeding
基金
国家 8 6 3计划 (项目编号 :2 0 0 1AA2 1110 1)
国家转基因植物专项 (项目编号 :J99-A - 0 0 9)
海南省重点科技项目 (项目编号 :0 12 0 2 )资助
关键词
大豆
孢囊线虫
4号生理小种
诱导表达
cDNA分析
合胞体
根系
Soybean (G. max), Soybean Cyst Nematode Race 4 (Heterodera glycines Ichinohe), mRNA Differential display (DDRT), cDNA analysis
