Resistance Reaction Of Rice Photo Thermo Sensitive Genic Male Sterile Lines To Southern Rice Black-Streaked Dwarf Virus

19rice photo thermo sensitive genic male sterile lines (PTGMS) were used to study the SRBSDV (Southern rice black streaked dwarf virus) identification method, their resistance to SRBSDV, the physiological change by SRBSDV infection, the inheritance, the NBS resistance genes. The main results were summarized as follows:1. The identification method of SRBSDV and resistantance evaluation of19rice sterile lines to SRBSDV:SRBSDV S10(EU523360) was used to design a pair of specific primers to detect SRBSDV, using molecular biology and the field investigation method, with reference to Guangdong province biological control experiment station’s grade standards about occurrence regularity and control technology research of southern black rice dwarf virus disease, we measured19rice sterile lines’plant height and disease grades, computing their average incidence, their disease index and their relative incidence. Clustal analysis with main component average incidence,19resistance of sterile lines are divided into four levels:(1) high resistant (HR):164S;(2) moderate resistant (MR):Fu3S, Shuguang1268S, Guangzhan63S;(3)moderate sensitive (MS):Ma S,6201S and95S;(4) high sensitive (HS):Huamo m207S, Huamo103S, An S, FSA,25S,116S,6303S, Huashi wy-2S, Chunghua38S, Anhui195S, Peiai64S and Chun S. Haven’t found any immune material.2. Physiological changes of rice by SRBSDV infection:(1) Analysis of the morphology and activities of ratooning rice root systems infected by SRBSDV:Three rice photo thermo sensitive genic male sterile lines which had been identified to be high resistant, moderate resistant and high sensitive to SRBSDV were used as materials and four disease grades (0,1,2and3grade) were set to study the influence of SRBSDV infection on sterile lines ratooning rice root morphology and activities by the method of hydroponic regeneration. The results showed that while there was no SRBSDV infection (grade0), root indexes reached significant differences between high resistant and high sensitive sterile lines, but there was no significant difference between high resistant and moderate resistant sterile lines. While sterile lines infected by SRBSDV (grade1,2,3), the root indexes indicated a downward trend as disease grades increased within the same sterile line. Indicators reached significant differences as the grades increased except the tips number of the high resistant sterile line. The influence of SRBSDV on root system mainly performed on different size roots.(2) Relationship analysis of endogenous hormone and PTGMS resistance to SRBSDVArtificial inoculating SRBSDV and HPLC (High Performance Liquid Chromatography) technique were used to measure the GA3, IAA, SA and ABA content of high resistant PTGMS line164S and high sensitive PTGMS line Peiai64S in response to SRBSDV infection. The results showed that:1) Peiai64S’GA3and IAA average content were significantly lower in SRBSDV infected plants than that in healthy plants. The lowest value appeared in the5th and6th day after SRBSDV infected when disease symptoms began to appear.164S’IAA average content was significantly lower in SRBSDV infected plants than those in healthy plants; GA3average content was lower in the infected plants than those in healthy plants, but didn’t reach the5%significant level.2) Resistance signal molecule SA content was higher in SRBSDV infected plants than in healthy plants. Both reached the highest level when disease symptoms initialised. Peiai64S’ SA content declined faster than that of164S’ after the highest value and164S’ SA content remained significantly higher than that in healthy plants.3) Endogenous hormone ABA which relevants to aging and death content was higher in SRBSDV infected plants than that in healthy plants. Peiai64S’content significantly higher than164S’ and significantly higher than their healthy plants;164S didn’t show significant difference with its healthy plants.4) SRBSDV infection changed plants’ IAA/ABA and GA3/ABA balance. Conclusion: SRBSDV stress changed rice endogenous hormone content, but this change was direct infection or indirect damage of the virus still remains elusive.(3) Establishment and implication of SRBSDV real-time qPCRDesign a pair of specific primers to amplify SRBSDV CP gene fragment, UBC was used as referance gene, establish SRBSDV CP two-step real time absolute quantitative PCR detection method. Absolute quantitative the grade0, grade1, grade2, grade3rice leaf SRBSDV CP copies number, through UBC regulation, results show that SRBSDV CP copies number significantly difference between grades.3. SRBSDV resistance genetic analysis of rice sterile lines: 3moderate resistant and1high resistant rice photo thermo sensitive genic male sterile line to SRBSDV and sensitive check TN1were used as materials in this study. Classical genetic analysis was applied to study this five SRBSDV resistant parent genetic and logarithm of resistance genes.164S hybrided with TN1, their F2hybrids showed resistant to sensitive15:1separation ratio. The result indicates164S’ resistance to SRBSDV is controlled by2dominant genes. Four sterile lines Guangzhan63S, Shuguang1268S, Fu3S hybrided with TN1, their F2hybrids showed resistant to sensitive15:1separation ratio. The results showed that their resistances to SRBSDV are controlled by1dominant gene.4. Rice sterile lines resistance genes homologous cloning and sequence analysis:Design a pair of degenerate primers according to the NBS-LRR conserved amino acid sequences, homologous cloning and analysis the rice sterile lines’ disease resistant gene sequences. The results showed that18rice sterile lines, Enhui58and TN1, received9types of NBS-LRR resistance gene homologous sequences total. These9types of resistance genes homologous amino acid sequence similarities are between49.5%～83.6%. Contrast with the9had been known NBS resistance genes amino acid sequences similarities are26.2%～100.0%.