南方根结线虫Col基因分子克隆及其VIGS效应分析

Molecular clone of Col gene in Meloidogyne incognita and analysis of its VIGS effect

为了明确线虫角质层胶原蛋白基因(Col)与根结线虫病害的关系,利用从线虫基因组中预测的Col设计特异引物,克隆了南方根结线虫Meloidogyne incognitacol基因MiCol。该基因完整编码区长903bp,编码300个氨基酸。克隆的MiCol与预测的MiCol基因序列一致性高达99.67%,氨基酸序列一致性高达100%。利用病毒介导的基因沉默技术,将其导入番茄植株并接种南方根结线虫,60 d后,沉默载体pTV-MiColi处理番茄植株的根结数比空载体对照及清水对照分别减少49.4%和49.2%,表明MiCol基因沉默显著降低了南方根结线虫的侵染数量。

In order to elucidate the relationship between nematode cuticle collagen gene （Col） and dis- ease caused by root-knot nematode, the MiCol gene in Meloidogyne incognita was cloned with special primers designed based on the predicted Col sequence in nematode genomes. The complete coding se- quence of the cloned MiCol was 903 bp encoding 300 amino acids. The identity between the cloned MiCol gene and the predicted MiCol gene was as high as 99.67% in nucleotide sequence and 100% in the de- duced amino acid sequence, respectively. Using virus-induced gene silence （VIGS） technique, the Mi- Col gene was transferred into tomato plantlets followed by inoculating with M. incognita. Sixty days later, the amounts of root knot on the tomato plantlets treated with the gene silencing vector pTV-MiColi de- creased by 49.4% or 49.2% compared with the empty vector control or water control, respectively. The study revealed that the MiCol gene silencing significantly reduced the infection of M. incognita