Sugarcane Yellow Leaf Virus Molecule Detection And ScYLV-CP Gene Clone

Sugarcane is the most important sugar crop in the world and is accounting for more than ninty perecent of Chinese sugar products. At the same time,sugarcane is an important crop for green energy production. The accumulation of virus in sugarcane stalks causes a yearly serious diseases because of vegetative propagation. Sugarcane Yellow Leaf Disease or Yellow Leaf Syndrome infected by Sugarcane Yellow Leaf Virus(ScYLV), is one of the most important sugarcane diseases in our country and in the world, causes severely decline in sugar content and stalk production of sugarcane. Breeding sugarcane varieties resistance to disease is the most effectively method to control the disease. Planting virus-free is another efficient method.Ten years is necessary to breeding a new variety by traditional crossing due to the complex genetic background. It is very difficult to breed a sugarcane variety resistant to Yellow Leaf Disease and with high stalk production and high sucrose content because of inadvantagous ligation of the genes controlly these characters. Due to the progress of plant genetic engineering, it is possible to improve the sugarcane disease resistance via transgene.Two pairs of specific primers are used in the research and the method of RT-PCR detection of ScYLV in sugarcane leaves are developed. One pair of primers named YLSR462 and YLSF111 is designed according to the Dr.Irey. Another pair of primers named YLSCP18 and YLSCP573 is designed by ourselves according to the amino acid conservative sequence of ScYLV recorded in GENBANK. Two core fragments of coat protein gene of ScYLV are isolated by RT-PCR. Two target bands after recovery and purification of the fragments. Recombinants are selected by colony coloi and endzyme digestion. The size of two fragments are 352bp and 556bp respectively by sequencing of ScYLV-CP gene. The results of sequence BLAST showed they are part sequences in coding region. Comparision the two fragments with the other reports of ScYLV-CP gene sequences, which are isolated by foreign researchers from strains with different geographic origins, the homology of amino acid sequence is over ninty-five percent. The results of this research provide the key technology for detection of ScYLV in sugarcane...