| Ningxia wolfberry (Lycium barbarum L.) is a plant belongs to Lycium L., Solanaceae, Solaneae Reichb.and Lyciinae Wettst.. The fruit of Lycium barbarum L. contains glycolipids, carotenoids, polyphenols, Lycium barbanan polysaccharide (LBP), several different vitamins, many microelements, etc., Our previous study showed that the quality of wolfberry decreased under doubled CO2 concentration treatment. Accordingly the present study is taken the point of view of plant physiology and molecular biology, physiology basis test methods, anew generation of high-throughput sequencing technology (Illumina HiSeq2500) and qRT-PCR technology methods to reveal the influence of Ningxia wolfberry fruit from different angles under doubled CO2 concentration. The specific results are as follows:1. By the determination of plant physiology methods for sucrose metabolism enzymatic activity of Ningxia wolfberry maturity fruit. The results showed that under high CO2 conentration, the activity of enzymes on sucrose synthesis and on sucrose cleavage of summer fruit (CO2 processing 90d) are that TR higher than CK, and invertase (Al, NI) activity was significantly different in this, while, in autumn fruit (CO2 process 120d), the activity of enzymes on sucrose synthesis and on sucrose cleavage are tower than summer fruit, to be specific, the activity of enzymes on sucrose cleavage are that TR higher than CK, the activity of enzymes on sucrose synthesis are that TR lower than CK, and SS(synthetic direction) was significantly different. To be concluded that the acid invertase(AI) and the neutral invertase(NI) activity might be the key factors of sucrose metabolism-related under doubled atmospheric CO2 concentrations.2. Taken Ningxia wolfberry maturity fruit samples CK (In open top chamber, the CO2 concentration is 350±20μmol·mol-1 in entire growth period), TR (In open top chamber, the COt concentration is 700±20μmol·mol-1 in entire growth period), CT (In open top chamber, the CO2 concentration is 350±20μmol·mol-1 in entire growth period) as materials and divided it into three groups (two as a biological replicates, divided into 3 groups)to extracted RNA. The transcriptome were obtained after sequencing 44.45Gb Clean Data, each sample Clean Data are up to 6.66Gb, Q30 base percentage at 86.38%and above. Overall, a higher rate of correct sequencing, sequencing results ideal for Ningxia wolfberry transcriptome studies provided a positive and reliable data. After De novo assembly to obtain a total of 81,100 pieces Unigene, annotation information obtained is 35,111, more than 1kb in length Unigene has 21,005 of all annotation information, accounting for 25.9% of overall Unigene. Therefore, this sample composition result was more ideal, provided a good basis for comparison of different groups between Ningxia wolfberry fruit samples under different CO2 concentrations. Analysised the structure of the resulting gene-based Unigene database, which received a total of 9,931 SSR markers, and also conducted a CDS forecast and SNP analysis.3. Compared Ningxia wolfberry maturity fruit sequencing results from different obtained pairwise groups, found differentially expressed genes (DEG)between CK and TR, the quantity of DEG was 73.55 was up-regulated and 18 was down-regulated in it. All of that involves eight kinds of metabolic pathways, that was starch and sucrose metabolism, phenylpropanoid biosynthesis, pyruvate metabolism etc. Under processing by qRT-PCR of partial differential gene from CK and TR, the test results prove the reliability of the transcriptome sequencing. |
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