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近年来,马铃薯受根结线虫的危害日益严重,造成减产、植株萎蔫、影响其商品性,并使其很容易受其他土传病害侵染。而不同马铃薯材料对根结线虫抗病性存在明显差异。因此,从分子角度挖掘和利用其本身所具备的抗病基因变得尤为重要。本研究采用电子克隆技术,搜索马铃薯EST数据库,拼接获得28 714条重叠群(contigs),以番茄、辣椒中已经克隆的根结线虫抗性基因作为探针,与所拼接的contigs进行比对及序列分析,共获得2条马铃薯根结线虫抗性基因的c DNA序列(contig_5164,contig_12247)。进一步采用生物信息学方法,对所得2条抗性基因序列进行了氨基酸组成和亲缘关系的分析。结果显示,contig_5164总长1 191 bp,推测编码206个氨基酸,等电点为4.77。Contig_12247总长892 bp,推测编码114个氨基酸,等电点为4.24。氨基酸组成均以亮氨酸最多,谷氨酸其次。两基因与番茄亲缘关系最近。随后对拼接所得contigs进行功能注释和GO分类,完成了对马铃薯中5类抗病保守结构域基因的挖掘。本研究的结果可为马铃薯根结线虫抗性基因进一步的研究与应用以及开发利用马铃薯抗性资源奠定基础。
In recent years, potato is more and more endangered by root-knot nematodes, which leads to the decrease of yield and the wilting of plants, affecting its commerciality and making it susceptible to other soil-borne diseases. However, there was a significant difference in disease resistance between different potato materials and root knot nematode. Therefore, it is particularly important to tap and utilize the disease-resistant genes that are possessed by them from a molecular point of view. In this study, we searched EST database of potato by using electronic cloning technique to obtain 28 714 contigs. The root knot nematode resistance genes cloned from tomato and pepper were used as probes to compare with the contigs A total of 2 cDNA sequences of conidial root knot nematode resistance genes (contig_5164, contig_12247) were obtained by sequence analysis. Further bioinformatics methods were used to analyze the amino acid composition and genetic relationship of the two resistance gene sequences obtained. The results showed that contig_5164 has a total length of 1 191 bp, which is deduced to encode 206 amino acids with an isoelectric point of 4.77. The total length of Contig_12247 is 892 bp, presumed to encode 114 amino acids with an isoelectric point of 4.24. Amino acid composition are most leucine, glutamic acid followed. The two genes and tomato genetic relationship recently. Then, the functional contigs of splicing were annotated and the GO classification was completed, and the excavation of five types of resistance conserved domain genes in potato was completed. The results of this study may lay a foundation for further research and application of potato root knot nematode resistance gene and development and utilization of potato resistance resources.