The Differential Expression Of Genes Related To The Resistance Of Jujube Witches Broom Induced By Phytoplasma

Chinnese jujube (Ziziphus jujuba Mill.), a precious gerplasm resources originated from china and a kind of important and specific fruit tree in China, belongs to family Rhamnaceae. Jujube witches' broom (JWB), caused by phytoplasma, is one the most serious and destructive disease of Chinese jujube. It is important to clone the resistant gene by using molecular biology and resistant resources. By the aid of cDNA-AFLP analysis, the differential expression of resistance gene for JWB induced by phytoplasma in resistant cultivar 'Xingguang' were studied taking sensitive cultivar as the comparison. The main results obtained were as following:1. 86 differential expression bands were gained in this research, 26 bands were related to the resistance of JWB, 14 bands were related to the sensitive of JWB, 18 bands were related to the phytoplasma inoculating. 28 bands were related to the cultivar.Only 20 out of 130 pairs primer were screened for cDNA-AFLP analysis and 86 polymorphic bands were amplified. A differential band of 400 bp were amplified at different stages of Xingguang from 75 days to 115 days after grafting induced by phytoplasma by the primer combinaion of Eaa/Mgt. A differential band of 250 bp were amplified at different stages of Xingguang and Zanhuangdazao infected by phytoplasma2. The inducing phases of resistance mechanism of 'Xingguang' was confirmed. The result of the diffrential expression bands at different stages of Xingguang from 55days to 115 days after grafting showed that the start-up stage of the resistance mechanism was from 55 days to 75 days after grafting, the stage of abundant-expression of the resistance mechanism was from 75 days to 115 days after grafting and the steady stage of the resistance mechanism was since 115 days after grafting.3. A sequene of 233bp was primary confirmed to be related to the resistance of Jujube witches' broom.Four differential expression patterns were reclaimed. Three sequenes of the length 399 bp,233 bp and 240 bp were obtained. The analysis of Blast in NCBI showed the frgment of 233 bp was homology to the Transcript derived fragments (TDFs) from wheat leaves inoculated by stripe rust fungus by 95%, and was homology to the gene related to stress response of cassava by 100%. So this sequene was primary confirmed to be related to the induced by phytoplasma.