ABSTRACT
In order to study the transcriptional differences of Citrus medica var. sarcodactylis at different developmental stages, we explored the genes regulating the biosynthesis of the effective components. In this study, Illumina Hiseq 4 000 high-throughput sequencing technology was used to sequence the transcriptome of C. medica var. sarcodactylis at different developmental stages, 121 235 unigenes were obtained with an average length of 2 434 bp, 3 379 different genes were obtained using DESeq screening, which mainly connected to biological processes such as signal transmission, biological regulation, and metabolic processes, and enriched in metabolic pathways such as starch, sucrose metabolism, phenylpropanoid biosynthesis, and flavonoid biosynthesis. Further dynamic comparison of biosynthesis related genes of active ingredients: the expression levels of PAL, CHI, CYP75B1, ZDS, 4CL and FLS gradually increased as the fruit turned from green to yellow; the expressions of COMT, F3H and CYP73A increased at first and then decreased; CCR, HCT and HRP were down-regulated whereas up-regulated. This study provides references for further excavation of key genes in the biosynthesis of active components, as well as biopathway analysis of active components for C. medica var. sarcodactylis.
Subject(s)
Citrus/genetics , Computational Biology , Fruit/genetics , Gene Expression Profiling , Gene Expression Regulation, Plant , High-Throughput Nucleotide Sequencing , TranscriptomeABSTRACT
Objective::To analyze and compare different samples in many aspects to identify Citrus medica var. sarcodactylis infected with Huanglongbing(HLB) timely and accurately, in order to prevent and control the disease in time. Method::HLB was identified through character analysis, reverse transcription-polymerase chain reaction(RT-PCR), enzyme digestion reaction and Real-time PCR. Result::In terms of characters, there were typically variegated yellow leaves and relatively small fruit, even with deformity but without " red nose fruit" among C. medica var. sarcodactylis infected with HLB. All of these can be used as the basis for the preliminary identification of HLB in the fields. According to the RT-PCR test results and enzyme digestion reaction, when the primer was OI1/OI2c, there was specific band of 1 160 bp, which could be cut into 520 bp and 640 bp by Xba I enzyme. These results were consistent with the characters of other citrus plants infected with HLB. According to the Real-time PCR detection results, C. medica var. sarcodactylis infected with HLB had amplification curves and dissolved peaks, with the melting temperature was 82 ℃ and Ct between 24.6 to 28.2, while the normal plants were not amplified. Conclusion::Character analysis can be used to roughly distinguish HLB in the fields, but with a certain subjectivity. RT-PCR or Real-time PCR can be used to identify C. medica var.sarcodactylis infected with HLB in a timely and accurate manner, and qPCR detection is more sensitive and quantitative. Through the combination of character analysis and molecular identification, C. medica var.sarcodactylis infected with HLB can be determined more timely and accurately.