Transcriptomic Analysis Of Sisal Infected By Phytophthora NicotianaeBreda

Sisal (Agave sisalana L.), the most important fiber crop in tropical regions of China. It possesses multiple usages, wide development foreground and high integrated values. Unfortunately, a kind of disease, named Zebra Disease, is one of the most common and serious disease in sisal. Caused by Phytophthora nicotianae., the Zebra Disease seriously affected the growth of sisal, cause hemp district deficiency in great quantities, led to the reduction of seedling in the early growth stage and eventually caused critical damage to yield of sisal.The pathogenic fungus of which, Phytophthora nicotianae., was diploid eukaryotic organisms(Chromista), traditional chitin sterilizing agent have no use in oomycetes. Currently, themolecular mechanisms of Sisal-Phytophthora nicotianae. interaction is unclear.In this study, we use Illuminahigh-throughput sequencing technology to analysis the transcriptome of sisal affected by Phytophthora nicotianae., use Real-time PCR, the semi-quantitative PCR to two-factor authentication, and then select the upregulated gene construct RNA interference vectors and transformed sisal plants. The main results are as follows:1. Three different methods were used toinoculated the Phytophthora nicotianae., to compare, we found that the most suitable method for Phytophthora nicotianae.Inoculation is homoiothermy moisturizing inoculation.2. Three different RNA extraction schemes indicated that improved sisal total RNA extracted kit havethe best quality.3. By transcriptome, we obtained131422non-redundant unigenes, total bases around8GB, most of fragment length between500to1000bp, the number of unigenes was length dependent, All-unigene N50is equal to the value of861, indicating that the ideal transcriptome sequencing results.4. By Nr blastn, we found that vitis vinifera have the most annotation,and most of then have40-80%similarities. There have507unigenes annotated to defense response by COG analysis, GO classification the42,987genes annotated to three ontology,KEGG analysis data then into128classes.5. Transcripts down-regulated genes more than up-regulated genes in which there are680Unigene pathogen interactions involved in the metabolic pathway, the expression fold difference from212to2-14, and we found that the leave of susceptible plants phenylalanine ammonia lyase gene expression more, surface plays an important role in the anti-zebra disease.6. Compaired by t ranscriptome sequencing、Real-time PCR and semi-quantitative PCR,we found that transcriptome sequencing data and reliable, for subsequent analysis.7.14RNAi vector were identified by sequencing and double enzyme identification, confirms the gateway cloning technology is a fast, efficient and accurate vector construction methods.