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以鸭梨为试材,采用qRT-PCR技术克隆鸭梨果实几丁质酶基因PbChi Ⅱ,并对几丁质酶氨基酸序列进行聚类分析;同时采用荧光定量PCR的方法对鸭梨植株不同器官及水杨酸(salicylic acid,SA)和梨轮纹病菌(Botryosphaeria berengeriana de.Not.f.sp.piricola(Nose)Koganecawa et.Sokwwa)处理后几丁质酶基因的表达量进行了分析,以期为该基因的进一步研究与利用提供参考依据。结果表明:PbChi Ⅱ基因长度为969bp,核苷酸序列及推导的氨基酸序列与沙梨的同源性均达到100%,该基因属于第Ⅱ类几丁质酶基因;PbChi Ⅱ在根和果实中表达量较大。在鸭梨果实中,SA和病原菌可诱导该基因表达,且表达量在48h达到最高,初步推测PbChi Ⅱ可能参与SA介导的植物抗病防卫反应的信号通路及轮纹病菌引起的防卫反应。
Using pear as test material, the chitinase gene PbChi Ⅱ was cloned by qRT-PCR and the amino acid sequence of chitinase was clustered. The fluorescence quantitative PCR And salicylic acid (SA) and Botryosphaeria berengeriana de.Not.f.sp.piricola (Nose) Koganecawa et. Sokwwa) were analyzed. Provide reference for the further study and utilization of this gene. The results showed that the length of PbChi Ⅱ gene was 969bp, the nucleotide sequence and the deduced amino acid sequence were all 100% homologous with Sapirudinosa. The gene belongs to class Ⅱ chitinase gene. PbChi Ⅱ is expressed in roots and fruits Larger amount. In Ya pear fruit, SA and pathogen could induce the expression of this gene, and the expression level reached the peak at 48h. It was preliminarily presumed that PbChi Ⅱ might be involved in the signal transduction pathways induced by SA in plant defense response and the defensive response caused by rotten germs.