Expressions of Defense Signal Pathway Genes in Tomato Plant Induced by Ralstonia solanacearum of Different Virulence
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摘要:
目的 探究不同致病力青枯雷尔氏菌诱导番茄防御相关基因的表达水平，为探明无致病力青枯雷尔氏菌的生防机制奠定基础。 方法 以无菌水为对照（CK），分别接种青枯雷尔氏菌强致病力菌株FJAT91和无致病力菌株FJAT1458，接种后不同时间（0 ，3，6 ，12 ，24 ，36 ，48 ，72 h）取样，利用荧光定量PCR技术，检测接种不同致病力青枯雷尔氏菌后番茄植株中水杨酸（SA）信号转导途径的gluA和PR-1a基因、茉莉酸（JA）信号转导途径的loxA和pin2基因及乙烯（ET）信号转导途径的Osm和PR-1a基因相对表达量的变化。 结果 菌株FJAT1458和FJAT91均能诱导番茄gluA、PR-1a、loxA、pin2、Osm和PR-1a基因的表达，接种后期（36 h之后），这些基因（PR-1a和Osm除外）在FJAT1458处理组的表达量均比FJAT91处理组高；FJAT1458和FJAT91诱导gluA和PR-1A基因表达量呈“先上升后下降”的趋势，接种12 h表达量最大；FJAT1458诱导番茄植株loxA和pin2基因表达量均显著高于CK，FJAT91诱导番茄植株loxA基因和pin2基因表达量在接种初期显著高于CK和FJAT1458处理组，后期迅速降低；FJAT-91诱导番茄植株Osm基因显著高于FJAT1458和对照处理（72 h除外），该菌株诱导番茄植株PR-1b基因表达量变化较为强烈，接种3 h，其诱导的PR-1b基因表达量为CK的251.33倍。 结论 不同致病力青枯雷尔氏菌诱导番茄防御相关基因表达量不同，可以进一步通过转录组法挖掘更多差异表达的基因。 Abstract:Objective Expressions of the defense-related genes in tomato plant induced by Ralstonia solanacearum of varied pathogenicity were studied to understand the biocontrol mechanisms of the avirulent FJAT1458. Method Real-time PCR was used to determine the expressions of gluA and PR-1a in the salicylic acid signal pathway, loxA and pin2 in the jasmonic acid signal pathway, and Osm and PR-1a in the ethylene signal pathway in tomato plants inoculated with the virulent strain FJAT91, the avirulent strain FJAT1458, or sterile water (CK) for 0, 3, 6, 12, 24, 36, 48, and 72 h. Result Both avirulent and virulent strains could induce expressions of gluA, PR-1a, loxA, pin2, Osm, and PR-1a. However, except for PR-1a and Osm, the gene expressions induced by FJAT1458 were significantly higher than those by FJAT91. Both FJAT1458 and FJAT91 inoculations raised the expressions of gluA and PR-1a to a peak followed by a decline in 12 h. The expressions of loxA and pin2 induced by FJAT1458 induction were significantly higher than CK, but FJAT91 did only initially. On the other hand, FJAT91 produced a significantly higher Osm expression than either FJAT1458 or CK, excluding the 72 h inoculation. And in the 3 h inoculation it rendered a PR-1b expression significantly stronger than FJAT1458 at 260.46 times higher than CK. Conclusion Both avirulent and virulent R. solanacearum could induce expressions of the defense-related genes in tomato plants in varied degrees. The transcriptomic method applied in this study could conceivably be used to unveil additional differently expressed genes associated with the pathways in tomato plants in the future. -
图 1 接种青枯菌不同时间番茄叶片gluA基因（A）和PR-1a基因（B）相对表达量
Figure 1. Relative expressions of gluA (A) and PR-1a (B) in tomato leaves after R. solanacearum inoculation for different time lengths
图 2 接种青枯菌不同时间番茄叶片loxA基因（A）和pin2基因（B）相对表达量
Figure 2. Relative expressions of loxA (A) and pin2 (B) in tomato leaves after R. solanacearum inoculation for different time lengths
图 3 接种青枯菌不同时间番茄叶片Osm基因（A）和PR-1b基因（B）相对表达量
Figure 3. Relative expressions of Osm (A) and PR-1b (B) in tomato leaves after R. solanacearum inoculation for different time lengths
表 1 RT-PCR检测基因的特异引物序列
Table 1. Specific primer sequences used for RT-PCR
检测基因
Tested
gene信号通路
Signal pathways基因库
登记号
GenBank accession no.引物序列
Primer sequencegIuA SA M80604 F: TCAGCAGGGTTGCAAAATCA R: CTCTAGGTGGGTAGGTGTTGGTTAA PR-1a SA M69247 F: TCT TGT GAG GCC CAA AAT TC R: ATA GTC TGG CCT CTC GGA CA loxA JA U09026 F: TGGTAGACCACCAACACGAA R: GACCAAAACGCTCGTCTCTC pin2 JA AY129402 F: TGATGCCAAGGCTTGTACTAGAGA R: AGCGGACTTCCTTCTGAACGT PR-1b ET X14065 F: CCA AGA CTA TCT TGC GGT TC R: GAA CCT AAG CCA CGA TAC CA Osmotin-like ET M21346 F: TGTACCACGTTTGGAGGACA R: ACCAGGGCAAGTAAATGTGC 
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